Search results for: green fluorescent protein

Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

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Authors: Oğuz Taşbozan, Celal Erbaş, Şefik Surhan Tabakoğlu, Mahmut Ali Gökçe

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Polyunsaturated fatty acids (PUFAs) and particularly the levels and ratios of ω-3 and ω-6 fatty acids are important for biological functions in humans and recognized as essential components of human diet. According to the terms of many different points of view, the nutritional composition of fish in culture conditions and caught from wild are wondered by the consumers. Therefore the aim of this study was to investigate the chemical composition of cage-cultured and wild sharpsnout seabream which has been preferred by the consumers as an economical important fish species in Turkey. The fish were caught from wild and obtained from cage-cultured commercial companies. Eight fish were obtained for each group, and their average weights of the samples were 245.8±13.5 g for cultured, 149.4±13.3 g for wild samples. All samples were stored in freezer (-18 °C) and analyses were carried out in triplicates, using homogenized boneless fish fillets. Proximate compositions (protein, ash, moisture and lipid) were determined. The fatty acid composition was analyzed by a GC Clarous 500 with auto sampler (Perkin–Elmer, USA). Proximate compositions of cage-cultured and wild samples of sharpsnout seabream were found statistical differences in terms of proximate composition between the groups. The saturated fatty acid (SFA), monounsaturated fatty acid (MUFA) and PUFA amounts of cultured and wild sharpsnout seabream were significantly different. ω3/ω6 ratio was higher in the cultured group. Especially in protein level and lipid level of cultured samples was significantly higher than wild counterparts. One of the reasons for this, cultured species exposed to continuous feeding. This situation had a direct effect on their body lipid content. The fatty acid composition of fish differs depending on a variety of factors including species, diet, environmental factors and whether they are farmed or wild. The higher levels of MUFA in the cultured fish may be explained with the high content of monoenoic fatty acids in the feed of cultured fish as in some other species. The ω3/ω6 ratio is a good index for comparing the relative nutritional value of fish oils. In our study, the cultured sharpsnout seabream appears to be better nutritious in terms of ω3/ω6. Acknowledgement: This work was supported by the Scientific Research Project Unit of the University of Cukurova, Turkey under grant no FBA-2016-5780.

Keywords: Diplodus puntazo, cage cultured, PUFA, fatty acid

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Authors: Mario Hanna Louis Hanna

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The use of CAD (pc Aided layout) generation is ubiquitous inside the structure, engineering and construction (AEC) industry. This has led to its inclusion in the curriculum of structure faculties in Nigeria as an important part of the training module. This newsletter examines the moral troubles involved in implementing CAD (pc Aided layout) content into the architectural training curriculum. Using current literature, this study begins with the advantages of integrating CAD into architectural education and the responsibilities of various stakeholders in the implementation process. It also examines issues related to the terrible use of records generation and the perceived bad effect of CAD use on design creativity. The use of a survey technique, information from the architecture department of Chukwuemeka Odumegwu Ojukwu Uli college changed into accumulated to serve as a case observe on how the problems raised have been being addressed. The object draws conclusions on what guarantees a hit moral implementation. Tens of millions of human beings around the sector suffer from hepatitis C, one of the international's deadliest sicknesses. Interferon (IFN) is a remedy alternative for patients with hepatitis C, but these treatments have their aspect outcomes. Our research targeted growing an oral small molecule drug that goals hepatitis C virus (HCV) proteins and has fewer facet effects. Our contemporary study targets to broaden a drug primarily based on a small molecule antiviral drug precise for the hepatitis C virus (HCV). Drug improvement and the use of laboratory experiments isn't always best high-priced, however also time-eating to behavior those experiments. instead, on this in silicon have a look at, we used computational strategies to recommend a particular antiviral drug for the protein domain names of discovered in the hepatitis C virus. This examines used homology modeling and abs initio modeling to generate the 3-D shape of the proteins, then figuring out pockets within the proteins. Proper lagans for pocket pills were advanced the usage of the de novo drug design method. Pocket geometry is taken into consideration while designing ligands. A few of the various lagans generated, a different for each of the HCV protein domains has been proposed.

Keywords: drug design, anti-viral drug, in-silicon drug design, Hepatitis C virus (HCV) CAD (Computer Aided Design), CAD education, education improvement, small-size contractor automatic pharmacy, PLC, control system, management system, communication.

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Authors: Kevin A. Droguett, Edwin G. Pérez, Denis Fuentealba, Margarita E. Aliaga, Angélica M. Fierro

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Supramolecular chemistry is a field of growing interest. Moreover, studying the formation of host-guest complexes between macrocycles and dyes is highly attractive due to their potential applications. Examples of the above are drug delivery, catalytic process, and sensing, among others. There are different dyes of interest in the literature; one example is the quinolinone derivatives. Those molecules have good optical properties and chemical and thermal stability, making them suitable for developing fluorescent probes. Secondly, several macrocycles can be seen in the literature. One example is the cucurbiturils. This water-soluble macromolecule family has a hydrophobic cavity and two identical carbonyl portals. Additionally, the thermodynamic analysis of those supramolecular systems could help understand the affinity between the host and guest, their interaction, and the main stabilization energy of the complex. In this work, two 7-(diethylamino) quinoline-2 (1H)-one derivative (QD1-2) and their interaction with cucurbit[7]uril (CB[7]) were studied from an experimental and in-silico point of view. For the experimental section, the complexes showed a 1:1 stoichiometry by HRMS-ESI and isothermal titration calorimetry (ITC). The inclusion of the derivatives on the macrocycle lends to an upward shift in the fluorescence intensity, and the pKa value of QD1-2 exhibits almost no variation after the formation of the complex. The thermodynamics of the inclusion complexes was investigated using ITC; the results demonstrate a non-classical hydrophobic effect with a minimum contribution from the entropy term and a constant binding on the order of 106 for both ligands. Additionally, dynamic molecular studies were carried out during 300 ns in an explicit solvent at NTP conditions. Our finding shows that the complex remains stable during the simulation (RMSD ~1 Å), and hydrogen bonds contribute to the stabilization of the systems. Finally, thermodynamic parameters from MMPBSA calculations were obtained to generate new computational insights to compare with experimental results.

Keywords: host-guest complexes, molecular dynamics, quinolin-2(1H)-one derivatives dyes, thermodynamics

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Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

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Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) system can facilitate targeted genome editing in organisms. Dual or single guide RNA (gRNA) can program the Cas9 nuclease to cut target DNA in particular areas; thus, introducing concise mutations either via error-prone non-homologous end-joining repairing or via incorporating foreign DNAs by homologous recombination between donor DNA and target area. In spite of high demand of such promising technology, developing a well-organized procedure in order for reliable mining of potential target sites for gRNAs in large genomic data is still challenging. Hence, we aimed to perform high-throughput detection of target sites by specific PAMs for not only common Streptococcus pyogenes (SpCas9) but also for Neisseria meningitides (NmCas9) CRISPR-Cas systems. Previous research confirmed the successful application of such RNA-guided Cas9 orthologs for effective gene targeting and subsequently genome manipulation. However, Cas9 orthologs need their particular PAM sequence for DNA cleavage activity. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of the target site for the two orthogonals of Cas9 protein, we created a reliable procedure to explore possible gRNA sequences. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. Finally, a complete list of all potential gRNAs along with their locations, strands, and PAMs sequence orientation can be provided for both SpCas9 as well as another potential Cas9 ortholog (NmCas9). The artificial design of potential gRNAs in a genome of interest can accelerate functional genomic studies. Consequently, the application of such novel genome editing tool (CRISPR/Cas technology) will enhance by presenting increased versatility and efficiency.

Keywords: CRISPR/Cas9 genome editing, gRNA mining, SpCas9, NmCas9

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Authors: Agnieszka Krzemińska, Katarzyna Świderek, Vicente Molinier, Piotr Paneth

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In order to understand in details the interactions between ligands and the enzyme isotope effects were studied between clinically used drugs that bind in the active site of Human Immunodeficiency Virus Reverse Transcriptase, HIV-1 RT, as well as triazole-based inhibitor that binds in the allosteric pocket of this enzyme. The magnitudes and origins of the resulting binding isotope effects were analyzed. Subsequently, binding isotope effect of the same triazole-based inhibitor bound in the active site were analyzed and compared. Together, these results show differences in binding origins in two sites of the enzyme and allow to analyze binding mode and place of newly synthesized inhibitors. Typical protocol is described below on the example of triazole ligand in the allosteric pocket. Triazole was docked into allosteric cavity of HIV-1 RT with Glide using extra-precision mode as implemented in Schroedinger software. The structure of HIV-1 RT was obtained from Protein Data Bank as structure of PDB ID 2RKI. The pKa for titratable amino acids was calculated using PROPKA software, and in order to neutralize the system 15 Cl- were added using tLEaP package implemented in AMBERTools ver.1.5. Also N-terminals and C-terminals were build using tLEaP. The system was placed in 144x160x144Å3 orthorhombic box of water molecules using NAMD program. Missing parameters for triazole were obtained at the AM1 level using Antechamber software implemented in AMBERTools. The energy minimizations were carried out by means of a conjugate gradient algorithm using NAMD. Then system was heated from 0 to 300 K with temperature increment 0.001 K. Subsequently 2 ns Langevin−Verlet (NVT) MM MD simulation with AMBER force field implemented in NAMD was carried out. Periodic Boundary Conditions and cut-offs for the nonbonding interactions, range radius from 14.5 to 16 Å, are used. After 2 ns relaxation 200 ps of QM/MM MD at 300 K were simulated. The triazole was treated quantum mechanically at the AM1 level, protein was described using AMBER and water molecules were described using TIP3P, as implemented in fDynamo library. Molecules 20 Å apart from the triazole were kept frozen, with cut-offs established on range radius from 14.5 to 16 Å. In order to describe interactions between triazole and RT free energy of binding using Free Energy Perturbation method was done. The change in frequencies from ligand in solution to ligand bounded in enzyme was used to calculate binding isotope effects.

Keywords: binding isotope effects, molecular dynamics, HIV, reverse transcriptase

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Authors: Raka Rahmatulloh, Mohammad Ilham Nugraha, Muhammad Ifan Fathurrahman

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The agricultural sector covers a wide area, one of them is livestock subsector that supply needs of the food source of animal protein. Animal protein is produced by the main livestock production such as meat, milk, eggs, etc. Besides the main production, livestock would produce metabolic residue, so called livestock wastes. Characteristics of livestock wastes can be either solid (feces), liquid (urine), and gas (methane) which turned out to be useful and has economical value when well-processed and well-controlled. Nowadays, this livestock wastes is considered as a source of pollutants, especially water pollution. If the source of pollutants used in an integrated way, it will have a positive impact on organic farming and a healthy environment. Management of livestock wastes can be integrated with the farming sector to the planting and caring that rely on fertilizers. Most Indonesian farmers still use chemical fertilizers, where the use of it in the long term will disturb the ecological balance of the environment. One of the main efforts is to use organic fertilizers instead of chemical fertilizer that conducted by the Faculty of Animal Husbandry, Padjadjaran University. The method is to use the solid waste of livestock and agricultural wastes into liquid organic fertilizer, feed additive, biogas and vermicompost through decomposition. The decomposition takes as long as 14 days including aeration and extraction process using water as a nutrients solvent media which contained in decomposes and disinfection media to release pathogenic microorganisms in decomposes. Liquid organic fertilizer has highly efficient for the farmers to have a ratio of carbon/nitrogen (C/N) 25/1 to 30/1 and neutral pH (6.5-7.5) which is good for plant growth. Feed additive may be given to improve the digestibility of feed so that substances can be easily absorbed by the body for production. Biogas contains methane (CH4), which has a high enough heat to produce electricity. Vermicompost is an overhaul of waste organic material that has excellent structure, porosity, aeration, drainage, and moisture holding capacity. Based on the case study above, an integrated livestock wastes management program strongly supports the Indonesian government in the achievement of sustainable livestock development.

Keywords: integrated, livestock wastes, organic fertilizer, sustainable livestock development

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Authors: Marta Fernández Batalla, José María Santamaría García, Maria Lourdes Jiménez Rodríguez, Roberto Barchino Plata, Adriana Cercas Duque, Enrique Monsalvo San Macario

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Background: Primary Care Nursing is taking more autonomy in clinical decisions. One of the most frequent therapies to solve is related to the problems of maintaining a sufficient supply of food. Nursing diagnoses related to food are addressed by the nurse-family and community as the first responsible. Objectives and interventions are set according to each patient. To improve the goal setting and the treatment of these care problems, a technological tool is developed to help nurses. Objective: To evaluate the computational tool developed to support the clinical decision in feeding problems. Material and methods: A cross-sectional descriptive study was carried out at the Meco Health Center, Madrid, Spain. The study population consisted of four specialist nurses in primary care. These nurses tested the tool on 30 people with ‘need for nutritional therapy’. Subsequently, the usability of the tool and the satisfaction of the professional were sought. Results: A simple and convenient computational tool is designed for use. It has 3 main entrance fields: age, size, sex. The tool returns the following information: BMI (Body Mass Index) and caloric consumed by the person. The next step is the caloric calculation depending on the activity. It is possible to propose a goal of BMI or weight to achieve. With this, the amount of calories to be consumed is proposed. After using the tool, it was determined that the tool calculated the BMI and calories correctly (in 100% of clinical cases). satisfaction on nutritional assessment was ‘satisfactory’ or ‘very satisfactory’, linked to the speed of operations. As a point of improvement, the options of ‘stress factor’ linked to weekly physical activity. Conclusion: Based on the results, it is clear that the computational tools of decision support are useful in the clinic. Nurses are not only consumers of computational tools, but can develop their own tools. These technological solutions improve the effectiveness of nutrition assessment and intervention. We are currently working on improvements such as the calculation of protein percentages as a function of protein percentages as a function of stress parameters.

Keywords: feeding behavior health, nutrition therapy, primary care nursing, technology assessment

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Authors: Jean De Dieu Ayabagabo, Paul A.Onjoro, Karubiu P. Migwi, Marie C. Dusingize

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This study investigated the effects of feed on milk yield and quality through feed monitoring and quality assessment, and the consequent enteric methane gas emissions from smallholder dairy farms in drier areas of Rwanda, using the Tier II approach for four seasons in three zones, namely; Mayaga and peripheral Bugesera (MPB), Eastern Savanna and Central Bugesera (ESCB), and Eastern plateau (EP). The study was carried out using 186 dairy cows with a mean live weight of 292 Kg in three communal cowsheds. The milk quality analysis was carried out on 418 samples. Methane emission was estimated using prediction equations. Data collected were subjected to ANOVA. The dry matter intake was lower (p<0.05) in the long dry season (7.24 Kg), with the ESCB zone having the highest value of 9.10 Kg, explained by the practice of crop-livestock integration agriculture in that zone. The Dry matter digestibility varied between seasons and zones, ranging from 52.5 to 56.4% for seasons and from 51.9 to 57.5% for zones. The daily protein supply was higher (p<0.05) in the long rain season with 969 g. The mean daily milk production of lactating cows was 5.6 L with a lower value (p<0.05) during the long dry season (4.76 L), and the MPB zone having the lowest value of 4.65 L. The yearly milk production per cow was 1179 L. The milk fat varied from 3.79 to 5.49% with a seasonal and zone variation. No variation was observed with milk protein. The seasonal daily methane emission varied from 150 g for the long dry season to 174 g for the long rain season (p<0.05). The rain season had the highest methane emission as it is associated with high forage intake. The mean emission factor was 59.4 Kg of methane/year. The present EFs were higher than the default IPPC value of 41 Kg from developing countries in African, the Middle East, and other tropical regions livestock EFs using Tier I approach due to the higher live weight in the current study. The methane emission per unit of milk production was lower in the EP zone (46.8 g/L) due to the feed efficiency observed in that zone. Farmers should use high-quality feeds to increase the milk yield and reduce the methane gas produced per unit of milk. For an accurate assessment of the methane produced from dairy farms, there is a need for the use of the Life Cycle Assessment approach that considers all the sources of emissions.

Keywords: footprint, forage, girinka, tier

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Authors: Laura Rueda-Gensini, Jader Rodríguez, Juan C. Cruz, Carolina Munoz-Camargo

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There is an emerging interest in botanicals and plant extracts for medicinal practices due to their widely reported health benefits. A large variety of phytochemicals found in plants have been correlated with antioxidant, immunomodulatory, and analgesic properties, which makes plant-derived products promising candidates for modulating the progression and treatment of numerous diseases. Non-centrifugal cane sugar (NCS), in particular, has been known for its high antioxidant and nutritional value, but composition-wise variability due to changing environmental and processing conditions have considerably limited its use in the nutraceutical and biomedical fields. This work is therefore aimed at assessing the effect of thermal exposure during NCS production over its bioactive composition and, in turn, its therapeutic value. Accordingly, two modified dehydration methods are proposed that employ: (i) vacuum-aided evaporation, which reduces the necessary temperatures to dehydrate the sample, and (ii) window refractance evaporation, which reduces thermal exposure time. The biochemical composition of NCS produced under these two methods was compared to traditionally-produced NCS by estimating their total polyphenolic and protein content with Folin-Ciocalteu and Bradford assays, as well as identifying the major phenolic compounds in each sample via HPLC-coupled mass spectrometry. Their antioxidant activities were also compared as measured by their scavenging potential of ABTS and DPPH radicals. Results show that the two modified production methods enhance polyphenolic and protein yield in resulting NCS samples when compared to traditional production methods. In particular, reducing employed temperatures with vacuum-aided evaporation demonstrated to be superior at preserving polyphenolic compounds, as evidenced both in the total and individual polyphenol concentrations. However, antioxidant activities were not significantly different between these. Although additional studies should be performed to determine if the observed compositional differences affect other therapeutic activities (e.g., anti-inflammatory, analgesic, and immunoprotective), these results suggest that reducing thermal exposure holds great promise for the production of natural products with enhanced nutritional value.

Keywords: non-centrifugal cane sugar, polyphenolic compounds, thermal processing, antioxidant activity

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Authors: Bekmurza H. Aitchanov, Sholpan K. Aitchanova, Olimzhon A. Baimuratov, Aitkul N. Aldibekova

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This work considers the automated control system (ACS) of milk quality during its magnetic field processing. For achieving high level of quality control methods were applied transformation of complex nonlinear systems in a linearized system with a less complex structure. Presented ACS is adjustable by seven parameters: mass fraction of fat, mass fraction of dry skim milk residues (DSMR), density, mass fraction of added water, temperature, mass fraction of protein, acidity.

Keywords: fluids magnetization, nuclear magnetic resonance, automated control system, dynamic pulse-frequency modulator, PFM, nonlinear systems, structural model

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Authors: Saeed Rezaeian, M. Rezaee Boroon

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Nitrogen accumulates in soils during the process of fertilizer addition to promote the plant growth. When the organic matter decomposes, the form of available nitrogen produced is in the form of nitrate, which is highly mobile. The most significant health effect of nitrate ingestion is methemoglobinemia in infants under six months of age (blue baby syndrome). The mobile nutrients, like nitrate nitrogen, are not stored in the soil as the available forms for the long periods and in large amounts. It depends on the needs for the crops such as vegetables. On the other hand, the vegetables will compete actively for nitrate nitrogen as a mobile nutrient and water. The mobile nutrients must be shared. The fewer the plants, the larger this share is for each plant. Also, this nitrate nitrogen is poisonous for the people who use these vegetables. Nitrate is converted to nitrite by the existing bacteria in the stomach and the Gastro-Intestinal (GI) tract. When nitrite is entered into the blood cells, it converts the hemoglobin to methemoglobin, which causes the anoxemia and cyanosis. The increasing use of pesticides and chemical fertilizers, especially the fertilizers with nitrates compounds, which have been common for the increased production of agricultural crops, has caused the nitrate pollution in the (soil, water, and environment). They have caused a lot of damage to humans and animals. In this research, the nitrate accumulation in different kind of vegetables such as; green pepper, tomatoes, egg plants, watermelon, cucumber, and red pepper were observed in the suburbs of Mashhad, Neisabour, and Sabzevar cities. In some of these cities, the information forms of agronomical practices collected were such as; different vegetable crops fertilizer recommendations, varieties, pesticides, irrigation schedules, etc., which were filled out by some of our colleagues in the research areas mentioned above. Analysis of the samples was sent to the soil and water laboratory in our department in Mashhad. The final results from the chemical analysis of samples showed that the mean levels of nitrates from the samples of the fruit crops in the mentioned cities above were all lower than the critical levels. These fruit crop samples were in the order of: 35.91, 8.47, 24.81, 6.03, 46.43, 2.06 mg/kg dry matter, for the following crops such as; tomato, cucumber, eggplant, watermelon, green pepper, and red pepper. Even though, this study was conducted with limited samples and by considering the mean levels, the use of these crops from the nutritional point of view will not cause the poisoning of humans.

Keywords: environmental pollution, human health, nitrate accumulations, nitrate fertilizers

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Authors: Sontana Mimapan, Phattarawadee Wattanasuntorn, Phanom Saijit

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Aflatoxin contamination in maize, one of several agriculture crops grown for livestock feeding, is still a problem throughout the world mainly under hot and humid weather conditions like Thailand. In this study Aspergillus flavus (A. Flavus), the key fungus for aflatoxin production especially aflatoxin B1 (AFB1), isolated from naturally infected maize were identified and characterized according to colony morphology and PCR using ITS, Beta-tubulin and calmodulin genes. The strains were analysed for the presence of four aflatoxigenic biosynthesis genes in relation to their capability to produce AFB1, Ver1, Omt1, Nor1, and aflR. Aflatoxin production was then confirmed using immunoaffinity column technique. A loop-mediated isothermal amplification (LAMP) was applied as an innovative technique for rapid detection of target nucleic acid. The reaction condition was optimized at 65C for 60 min. and calcein flurescent reagent was added before amplification. The LAMP results showed clear differences between positive and negative reactions in end point analysis under daylight and UV light by the naked eye. In daylight, the samples with AFB1 producing A. Flavus genes developed a yellow to green color, but those without the genes retained the orange color. When excited with UV light, the positive samples become visible by bright green fluorescence. LAMP reactions were positive after addition of purified target DNA until dilutions of 10⁻⁶. The reaction products were then confirmed and visualized with 1% agarose gel electrophoresis. In this regards, 50 maize samples were collected from dairy farms and tested for the presence of four aflatoxigenic biosynthesis genes using LAMP technique. The results were positive in 18 samples (36%) but negative in 32 samples (64%). All of the samples were rechecked by PCR and the results were the same as LAMP, indicating 100% specificity. Additionally, when compared with the immunoaffinity column-based aflatoxin analysis, there was a significant correlation between LAMP results and aflatoxin analysis (r= 0.83, P < 0.05) which suggested that positive maize samples were likely to be a high- risk feed. In conclusion, the LAMP developed in this study can provide a simple and rapid approach for detecting AFB1 producing A. Flavus genes from maize and appeared to be a promising tool for the prediction of potential aflatoxigenic risk in livestock feedings.

Keywords: Aflatoxin B1, Aspergillus flavus genes, maize, loop-mediated isothermal amplification

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Authors: Marcel Ciesla, Victoria Oberascher, Sven Eder, Stefan Kirchweger, Wolfgang E. Baaske, Gerald Ostermayer

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The omnipresent trend towards sustainable mobility is a major challenge, especially for commuters in rural areas. The use of micro public transport systems is expected to significantly reduce pollutant emissions, as several commuters travel the first mile together with a single pick-up bus instead of their own car. In this paper, different aspects of such a micro public transport system are analyzed. The main findings of the investigations should be how the travel times of commuters change and how many CO₂ emissions can be saved if some of the commuters use public transport instead of their own vehicle.

Keywords: micro public transport, green transportation, sustainable mobility, low-emission commuting

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Authors: Nseabasi Nsikakabasi Etim

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In recent times, there has been a significant shortfall in between the production and supply of animal protein to meet the ever increasing population. To meet the increasing demand for animal protein, there is a need to focus attention on the production of livestock whose nutritional requirement does not put much strain on the limited sources of feed ingredients to which men subscribe. An example of such livestock is the rabbit. Rabbit is a pseudo-ruminant herbivore which utilizes much undigested and unabsorbed feed materials as sources of nutrient for maintenance and production. Thus, this study was conducted to investigate the effects of feeding Aspilia africana as forage on the growth rates of female pseudo-ruminant herbivores (rabbits) at different physiological states. Thirty (30) Dutch breed rabbit does of 5–6 months of age were used for the experiment which was conducted in a completely randomized design for four months. The rabbits were divided into three treatment groups, ten does per treatment group; which consisted of mixed forages (Centrosema pubescent (200g), Panicum maximum (200g) and Ipomea batatas leaves (100g) without Aspilia africana (T1; control), fresh Aspilia africana (500g/dose/day) (T2) and wilted Aspilia africana (500g/dose/day) (T3). Rabbits in all treatment groups received the same concentrate (300g/animal/day) throughout the period of the study and mixed forages from the commencement of the experiment till the does kindled. After parturition, fresh and wilted Aspilia africana were introduced in treatments 2 and three respectively, whereas the control group continued on mixed forages throughout the study. The result of the study revealed that the initial average body weight of the rabbit does was 1.74kg. At mating and gestation periods, the body weights of the does in T2 was significantly higher (P<0.05) than the rest. There were no significant differences (P<0.05) in the body weights of does at kindling between the various treatment groups. During the physiological states of lactation, weaning and re-mating, the control group (T1) had significantly lower body weight than those of the treated groups (T2 and T3). Furthermore, T2 had significantly higher body weight than T3. The study revealed that Aspilia africana; mainly the fresh leaves have greater growth stimulating effects when fed to pseudo-ruminants (rabbits), thereby enhancing body weights of does during lactation and weaning.

Keywords: Aspilia africana, herbivores, pseudo-ruminants, physiological states

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Authors: Turban Kar, Maitree Bhattacharyya

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Curcumin, a dietary antioxidant has been identified as a wonder molecule to possess therapeutic properties protecting the cellular macromolecules from oxidative damage. In our experimental study, we have explored the effectiveness of curcumin in protecting the structural paradigm of Human Serum Albumin (HSA) when exposed to gamma irradiation. HSA, being an important transport protein of the circulatory system, is involved in binding of variety of metabolites, drugs, dyes and fatty acids due to the presence of hydrophobic pockets inside the structure. HSA is also actively involved in the transportation of drugs and metabolites to their targets, because of its long half-life and regulation of osmotic blood pressure. Gamma rays, in its increasing concentration, results in structural alteration of the protein and superoxide radical generation. Curcumin, on the other hand, mitigates the damage, which has been evidenced in the following experiments. Our study explores the possibility for protection by curcumin during the molecular and conformational changes of HSA when exposed to gamma irradiation. We used a combination of spectroscopic methods to probe the conformational ensemble of the irradiated HSA and finally evaluated the extent of restoration by curcumin. SDS - PAGE indicated the formation of cross linked aggregates as a consequence of increasing exposure of gamma radiation. CD and FTIR spectroscopy inferred significant decrease in alpha helix content of HSA from 57% to 15% with increasing radiation doses. Steady state and time resolved fluorescence studies complemented the spectroscopic measurements when lifetime decay was significantly reduced from 6.35 ns to 0.37 ns. Hydrophobic and bityrosine study showed the effectiveness of curcumin for protection against radiation induced free radical generation. Moreover, bityrosine and hydrophobic profiling of gamma irradiated HSA in presence and absence of curcumin provided light on the formation of ROS species generation and the protective (magical) role of curcumin. The molecular mechanism of curcumin protection to HSA from gamma irradiation is yet unknown, though a possible explanation has been proposed in this work using Thioflavin T assay. It was elucidated, that when HSA is irradiated at low dose of gamma radiation in presence of curcumin, it is capable of retaining the native characteristic properties to a greater extent indicating stabilization of molecular structure. Thus, curcumin may be utilized as a therapeutic strategy to protect cellular proteins.

Keywords: Bityrosine content, conformational change, curcumin, gamma radiation, human serum albumin

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Authors: Benedicte Madon, Marion Pillet, Justine Castrec, Quentin Fonatine, Pierre Lejeune, Michel Marengo, Helene Thomas

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Biodiversity in port ecosystems faces many anthropic pressures from port activities. The maritime industry and port areas have been under scrutiny regarding their environmental impacts. In the port value chain, port managers need to implement actions to fulfil environmental certifications and European Directive requirements. This paper seeks to highlight the lessons learned and opportunities through the QUAMPO project to move towards port biodiversity restoration in Corsica using innovative biomonitoring in the goal of obtaining green certification.

Keywords: biomonitoring, port, water quality, invertebrate, corsica, biomarker, trace elements, HAP, PCB, certification

Procedia PDF Downloads 117

Authors: Justus Enninga

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Enthusiasts and skeptics about economic growth have not much in common in their preference for institutional arrangements that solve ecological conflicts. This paper argues that agreement between both opposing schools can be found in the Bloomington Schools’ concept of polycentricity. Growth-enthusiasts who will be referred to as Simons after the economist Julian Simon and growth-skeptics named Ehrlichs after the ecologist Paul R. Ehrlich both profit from a governance structure where many officials and decision structures are assigned limited and relatively autonomous prerogatives to determine, enforce and alter legal relationships. The paper advances this argument in four steps. First, it will provide clarification of what Simons and Ehrlichs mean when they talk about growth and what the arguments for and against growth-enhancing or degrowth policies are for them and for the other site. Secondly, the paper advances the concept of polycentricity as first introduced by Michael Polanyi and later refined to the study of governance by the Bloomington School of institutional analysis around the Nobel Prize laureate Elinor Ostrom. The Bloomington School defines polycentricity as a non-hierarchical, institutional, and cultural framework that makes possible the coexistence of multiple centers of decision making with different objectives and values, that sets the stage for an evolutionary competition between the complementary ideas and methods of those different decision centers. In the third and fourth parts, it is shown how the concept of polycentricity is of crucial importance for growth-enthusiasts and growth-skeptics alike. The shorter third part demonstrates the literature on growth-enhancing policies and argues that large parts of the literature already accept that polycentric forms of governance like markets, the rule of law and federalism are an important part of economic growth. Part four delves into the more nuanced question of how a stagnant steady-state economy or even an economy that de-grows will still find polycentric governance desirable. While the majority of degrowth proposals follow a top-down approach by requiring direct governmental control, a contrasting bottom-up approach is advanced. A decentralized, polycentric approach is desirable because it allows for the utilization of tacit information dispersed in society and an institutionalized discovery process for new solutions to the problem of ecological collective action – no matter whether you belong to the Simons or Ehrlichs in a green political economy.

Keywords: degrowth, green political theory, polycentricity, institutional robustness

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Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

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Authors: Kriti Jain, Sucha Singh Gill

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Subsidies have been a major part of agricultural policies around the world, and more extensively since the green revolution in developing countries, for the sake of attaining higher agricultural productivity and achieving food security. But entrenched subsidies lead to distorted incentives and promote inefficiencies in the agricultural sector, threatening the viability of these very subsidies and sustainability of the agricultural production systems, posing a threat to the livelihood of farmers and laborers dependent on it. This paper analyzes the economic and ecological sustainability implications of prolonged input and output subsidies in agriculture by studying the case of Indian Punjab, an agriculturally developed state responsible for ensuring food security in the country when it was facing a major food crisis. The paper focuses specifically on the environmentally unsustainable cropping pattern changes as a result of Minimum Support Price (MSP) and assured procurement and on the resource use efficiency and cost implications of power subsidy for irrigation in Punjab. The study is based on an analysis of both secondary and primary data sources. Using secondary data, a time series analysis was done to capture the changes in Punjab’s cropping pattern, water table depth, fertilizer consumption, and electrification of agriculture. This has been done to examine the role of price and output support adopted to encourage the adoption of green revolution technology in changing the cropping structure of the state, resulting in increased input use intensities (especially groundwater and fertilizers), which harms the ecological balance and decreases factor productivity. Evaluation of electrification of Punjab agriculture helped evaluate the trend in electricity productivity of agriculture and how free power imposed further pressure on the extant agricultural ecosystem. Using data collected from a primary survey of 320 farmers in Punjab, the extent of wasteful application of groundwater irrigation, water productivity of output, electricity usage, and cost of irrigation driven electricity subsidy to the exchequer were estimated for the dominant cropping pattern amongst farmers. The main findings of the study revealed how because of a subsidy has driven agricultural framework, Punjab has lost area under agro climatically suitable and staple crops and moved towards a paddy-wheat cropping system, that is gnawing away the state’s natural resources like water table has been declining at a significant rate of 25 cms per year since 1975-76, and excessive and imbalanced fertilizer usage has led to declining soil fertility in the state. With electricity-driven tubewells as the major source of irrigation within a regime of free electricity and water-intensive crop cultivation, there is both wasteful application of irrigation water and electricity in the cultivation of paddy crops, burning an unproductive hole in the exchequer’s pocket. There is limited access to both agricultural extension services and water-conserving technology, along with policy imbalance, keeping farmers in an intensive and unsustainable production system. Punjab agriculture is witnessing diminishing returns to factor, which under the business-as-usual scenario, will soon enter the phase of negative returns to factor.

Keywords: cropping pattern, electrification, subsidy, sustainability

Procedia PDF Downloads 184

Authors: Elvan G. Bulut, Betul Goksun, Tugba G. Gun, Ozge Sakiyan Demirkol, Kamuran Ayhan, Kezban Candogan

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Dysphagia, difficulty in swallowing solid foods and thin liquids, is one of the common health threats among the elderly who require foods with modified texture in their diet. Although there are some commercial food formulations or hydrocolloids to thicken the liquid foods for dysphagic individuals, there is still a need for developing and offering new food products with enriched nutritional, textural and sensory characteristics to safely nourish these patients. 3D food printing is an appealing alternative in creating personalized foods for this purpose with attractive shape, soft and homogenous texture. In order to modify texture and prevent phase separation, hydrocolloids are generally used. In our laboratory, an optimized 3D printed beef-based formulation specifically for people with swallowing difficulties was developed based on the research project supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK Project # 218O017). The optimized formulation obtained from response surface methodology was 60% beef powder, 5.88% gelatin, and 0.74% kappa-carrageenan (all in a dry basis). This product was enriched with powders of freeze-dried beet, celery, and red capia pepper, butter, and whole milk. Proximate composition (moisture, fat, protein, and ash contents), pH value, CIE lightness (L*), redness (a*) and yellowness (b*), and color difference (ΔE*) values were determined. Counts of total mesophilic aerobic bacteria (TMAB), lactic acid bacteria (LAB), mold and yeast, total coliforms were conducted, and detection of coagulase positive S. aureus, E. coli, and Salmonella spp. were performed. The 3D printed products had 60.11% moisture, 16.51% fat, 13.68% protein, and 1.65% ash, and the pH value was 6.19, whereas the ΔE* value was 3.04. Counts of TMAB, LAB, mold and yeast and total coliforms before and after 3D printing were 5.23-5.41 log cfu/g, < 1 log cfu/g, < 1 log cfu/g, 2.39-2.15 log EMS/g, respectively. Coagulase positive S. aureus, E. coli, and Salmonella spp. were not detected in the products. The data obtained from this study based on determining some important product characteristics of functional beef-based formulation provides an encouraging basis for future research on the subject and should be useful in designing mass production of 3D printed products of similar composition.

Keywords: beef, dysphagia, product characteristics, texture-modified foods, 3D food printing

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Authors: Abdullah Al Mohiuddin

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Bangladesh is one of the poorest countries in the world, and ranks low on almost all measures of economic development, thus leaving the population extremely vulnerable to natural disasters and climate events. 20% of GDP come from agriculture but more than 60% of the population relies on agriculture as their main source of income making the entire economy vulnerable to climate change and natural disasters. High population density exacerbates the exposure to and effect of climate events, and increases the levels of vulnerability, as does the poor institutional development of the country. The most vulnerable sectors to climate change impacts in Bangladesh are agriculture, coastal zones, water resources, forestry, fishery, health, biomass, and energy. High temperatures, heavy rainfall, high humidity and fairly marked seasonal variations characterize the climate in Bangladesh: Mild winter, hot humid summer and humid, warm rainy monsoon. Much of the country is flooded during the summer monsoon. The Department of Environment (DOE) under the Ministry of Environment and Forestry (MoEF) is the focal point for the United Nations Framework Convention on Climate Change (UNFCCC) and coordinates climate related activities in the country. Recently, a Climate Change Cell (CCC) has been established to address several issues including adaptation to climate change. The climate change focus started with The National Environmental Management Action Plan (NEMAP) which was prepared in 1995 in order to initiate the process to address environmental and climate change issues as long-term environmental problems for Bangladesh. Bangladesh was one of the first countries to finalise a NAPA (Preparation of a National Adaptation Plan of Action) which addresses climate change issues. The NAPA was completed in 2005, and is the first official initiative for mainstreaming adaptation to national policies and actions to cope with climate change and vulnerability. The NAPA suggests a number of adaptation strategies, for example: - Providing drinking water to coastal communities to fight the enhanced salinity caused by sea level rise, - Integrating climate change in planning and design of infrastructure, - Including climate change issues in education, - Supporting adaptation of agricultural systems to new weather extremes, - Mainstreaming CCA into policies and programmes in different sectors, e.g. disaster management, water and health, - Dissemination of CCA information and awareness raising on enhanced climate disasters, especially in vulnerable communities. Bangladesh has geared up its environment conservation steps to save the world’s poorest countries from the adverse effects of global warming. Now it is turning towards green economy policies to save the degrading ecosystem. Bangladesh is a developing country and always fights against Natural Disaster. At the same time we also fight for establishing ecological environment through promoting Green Economy/Energy by Youth Networking. ANTAR is coordinating a big Youth Network in the southern part of Bangladesh where 30 Youth group involved. It can be explained as the economic development based on sustainable development which generates growth and improvement in human’s lives while significantly reducing environmental risks and ecological scarcities. Green economy in Bangladesh promotes three bottom lines – sustaining economic, environment and social well-being.

Keywords: resilience, networking, mobilizing, resource

Procedia PDF Downloads 308

Authors: Alice Nwanyioma Ohuoba, Rose Erdoo Kukwa, Ukpabi Joseph Ukpabi

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Cassava root (Manihot esculenta) is one of the important carbohydrates containing crops in Nigeria. It is a staple food, mostly in the southern part of the country, and a source of income to farmers and processors. Cassava gari processing methods result to residue fiber (solid waste) from the sieving operation, these residue fibers ( solid wastes) can be dried and milled into flour and used to prepare cakes, cookies, crackers and chin-chin instead of being thrown away mostly on farmland or near the residential area. Flour for baking or frying may contain carbohydrates and protein (wheat flour) or rich in only carbohydrates (cassava flour). Cake, cookies, crackers, and chin-chin were prepared using the residue flour obtained from the residue fiber of cassava variety NR87184 roots, processed into gari. This study is aimed at evaluating the proximate composition, mineral content and sensory attributes of these selected snacks produced. The proximate composition results obtained showed that crackers had the lowest value in moisture (2.3390%) and fat (1.7130%), but highest in carbohydrates (85.2310%). Amongst the food products, cakes recorded the highest value in protein (8.0910%). Crude fibre values ranges from 2.5265% (cookies) to 3.4165% (crackers). The result of the mineral contents showed cookies ranking the highest in Phosphorus (65.8535 ppm) and Iron (0.1150 mg/L), Calcium (1.3800mg/L) and Potassium (7.2850 mg/L) contents, while chin-chin and crackers were lowest in Sodium ( 2.7000 mg/L). The food products were also subjected to sensory attributes evaluation by thirty member panelists using 9-hedonic scale which ranged from 1 ( dislike extremely) to 9 (like extremely). The means score obtained shows all the food products having above 7.00 (above “like moderately”). This study has shown that food products that may be functional or nutraceuticals could be prepared from the residue flour. There is a call for the use of gluten-free flour in baking due to ciliac disease and other allergic causes by gluten. Therefore local carbohydrates food crops like cassava residue flour that are gluten-free, could be the solution. In addition, this could aid cassava gari processing waste management thereby reducing post-harvest losses of cassava root.

Keywords: allergy, flour, food-products, gluten-free

Procedia PDF Downloads 155

Authors: Valencia Fernandes, Dharmendra Kumar Khatri, Shashi Bala Singh

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Background and Aim: Epigenetics are the inaudible signatures of several pathological processes in the brain. This study understands the influence of DNA methylation, a major epigenetic modification, in the prefrontal cortex and hippocampus of the diabetic brain and its notable effect on the cellular chaperones and synaptic proteins. Method: Chronic high fat diet and STZ-induced diabetic mice were studied for cognitive dysfunction, and global DNA methylation, as well as DNA methyltransferase (DNMT) activity, were assessed. Further, the cellular chaperones and synaptic proteins were examined using DNMT inhibitor, 5-aza-2′-deoxycytidine (5-aza-dC)-via intracerebroventricular injection. Moreover, % methylation of these synaptic proteins were also studied so as to correlate its epigenetic involvement. Computationally, its interaction with the DNMT enzyme were also studied using bioinformatic tools. Histological studies for morphological alterations and neuronal degeneration were also studied. Neurogenesis, a characteristic marker for new learning and memory formation, was also assessed via the BrdU staining. Finally, the most important behavioral studies, including the Morris water maze, Y maze, passive avoidance, and Novel object recognition test, were performed to study its cognitive functions. Results: Altered global DNA methylation and increased levels of DNMTs within the nucleus were confirmed in the cortex and hippocampus of the diseased mice, suggesting hypermethylation at a genetic level. Treatment with AzadC, a global DNA demethylating agent, ameliorated the protein and gene expression of the cellular chaperones and synaptic fidelity. Furthermore, the methylation analysis profile showed hypermethylation of the hsf1 protein, a master regulator for chaperones and thus, confirmed the epigenetic involvement in the diseased brain. Morphological improvements and decreased neurodegeneration, along with enhanced neurogenesis in the treatment group, suggest that epigenetic modulations do participate in learning and memory. This is supported by the improved behavioral test battery seen in the treatment group. Conclusion: DNA methylation could possibly accord in dysregulating the memory-associated proteins at chronic stages in type 2 diabetes. This could suggest a substantial contribution to the underlying pathophysiology of several metabolic syndromes like insulin resistance, obesity and also participate in transitioning this damage centrally, such as cognitive dysfunction.

Keywords: epigenetics, cognition, chaperones, DNA methylation

Procedia PDF Downloads 202

Authors: Andreia Garrido, Artur Conde, Ana Cunha, Ric De Vos

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Climate changes predictions point to increases in abiotic stress for crop plants in Portugal, like pronounced temperature variation and decreased precipitation, which will have negative impact on grapevine physiology and consequently, on grape berry and wine quality. Short-term mitigation strategies have, therefore, been implemented to alleviate the impacts caused by adverse climatic periods. These strategies include foliar application of kaolin, an inert mineral, which has radiation reflection proprieties that decreases stress from excessive heat/radiation absorbed by its leaves, as well as smart irrigation strategies to avoid water stress. However, little is known about the influence of these mitigation measures on grape berries, neither on the photosynthetic activity nor on the photosynthesis-related metabolic profiles of its various tissues. Moreover, the role of fruit photosynthesis on berry quality is poorly understood. The main objective of our work was to assess the effects of kaolin and irrigation treatments on the photosynthetic activity of grape berry tissues (exocarp and seeds) and on their global metabolic profile, also investigating their possible relationship. We therefore collected berries of field-grown plants of the white grape variety Alvarinho from two distinct microclimates, i.e. from clusters exposed to high light (HL, 150 µmol photons m⁻² s⁻¹) and low light (LL, 50 µmol photons m⁻² s⁻¹), from both kaolin and non-kaolin (control) treated plants at three fruit developmental stages (green, véraison and mature). Plant irrigation was applied after harvesting the green berries, which also enabled comparison of véraison and mature berries from irrigated and non-irrigated growth conditions. Photosynthesis was assessed by pulse amplitude modulated chlorophyll fluorescence imaging analysis, and the metabolite profile of both tissues was assessed by complementary metabolomics approaches. Foliar kaolin application resulted in, for instance, an increased photosynthetic activity of the exocarp of LL-grown berries at green developmental stage, as compared to the control non-kaolin treatment, with a concomitant increase in the levels of several lipid-soluble isoprenoids (chlorophylls, carotenoids, and tocopherols). The exocarp of mature berries grown at HL microclimate on kaolin-sprayed non-irrigated plants had higher total sugar levels content than all other treatments, suggesting that foliar application of this mineral results in an increased accumulation of photoassimilates in mature berries. Unbiased liquid chromatography-mass spectrometry-based profiling of semi-polar compounds followed by ASCA (ANOVA simultaneous component analysis) and ANOVA statistical analysis indicated that kaolin had no or inconsistent effect on the flavonoid and phenylpropanoid composition in both seed and exocarp at any developmental stage; in contrast, both microclimate and irrigation influenced the level of several of these compounds depending on berry ripening stage. Overall, our study provides more insight into the effects of mitigation strategies on berry tissue photosynthesis and phytochemistry, under contrasting conditions of cluster light microclimate. We hope that this may contribute to develop sustainable management in vineyards and to maintain grape berries and wines with high quality even at increasing abiotic stress challenges.

Keywords: climate change, grape berry tissues, metabolomics, mitigation strategies

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Authors: Nahush Modak, Meena Pangarkar, Anand Pathak, Ankita Tamhane

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Background: Breast cancer is the prominent cause of cancer and mortality among women. This study was done to show the statistical analysis of a cohort of over 250 patients detected with breast cancer diagnosed by oncologists using Immunohistochemistry (IHC). IHC was performed by using ER; PR; HER2; Ki-67 antibodies. Materials and methods: Formalin fixed Paraffin embedded tissue samples were obtained by surgical manner and standard protocol was followed for fixation, grossing, tissue processing, embedding, cutting and IHC. The Ventana Benchmark XT machine was used for automated IHC of the samples. Antibodies used were supplied by F. Hoffmann-La Roche Ltd. Statistical analysis was performed by using SPSS for windows. Statistical tests performed were chi-squared test and Correlation tests with p<.01. The raw data was collected and provided by National Cancer Insitute, Jamtha, India. Result: Luminal B was the most prevailing molecular subtype of Breast cancer at our institute. Chi squared test of homogeneity was performed to find equality in distribution and Luminal B was the most prevalent molecular subtype. The worse prognostic indicator for breast cancer depends upon expression of Ki-67 and her2 protein in cancerous cells. Our study was done at p <.01 and significant dependence was observed. There exists no dependence of age on molecular subtype of breast cancer. Similarly, age is an independent variable while considering Ki-67 expression. Chi square test performed on Human epidermal growth factor receptor 2 (HER2) statuses of patients and strong dependence was observed in percentage of Ki-67 expression and Her2 (+/-) character which shows that, value of Ki depends upon Her2 expression in cancerous cells (p<.01). Surprisingly, dependence was observed in case of Ki-67 and Pr, at p <.01. This shows that Progesterone receptor proteins (PR) are over-expressed when there is an elevation in expression of Ki-67 protein. Conclusion: We conclude from that Luminal B is the most prevalent molecular subtype at National Cancer Institute, Jamtha, India. There was found no significant correlation between age and Ki-67 expression in any molecular subtype. And no dependence or correlation exists between patients’ age and molecular subtype. We also found that, when the diagnosis is Luminal A, out of the cohort of 257 patients, no patient shows >14% Ki-67 value. Statistically, extremely significant values were observed for dependence of PR+Her2- and PR-Her2+ scores on Ki-67 expression. (p<.01). Her2 is an important prognostic factor in breast cancer. Chi squared test for Her2 and Ki-67 shows that the expression of Ki depends upon Her2 statuses. Moreover, Ki-67 cannot be used as a standalone prognostic factor for determining breast cancer.

Keywords: breast cancer molecular subtypes , correlation, immunohistochemistry, Ki-67 and HR, statistical analysis

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Authors: M. H. Moreira da Silva, L. Valadao, F. Moreira da Silva

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In the present study, the fertilizing capacity of bovine spermatozoa was evaluated before and after its cryopreservation. For this, the testicles of 100 bulls slaughtered on Terceira Island were dissected, the epididymal tails were separated, and semen was recovered by the flotation method and then evaluated by phase contrast microscopy and by flow cytometry. For phase contrast microscopy, a drop of semen was used to evaluate the percentage of motile spermatozoa (from 0 to 100%) and motility (from 0 to 5). After determining the concentration and the abnormal forms, semen was diluted to a final concentration of 50 x 106 spz/ml and evaluated by flow cytometer for membrane and acrosome integrity using the conjugation of fluorescent probes propidium iodide (PI) and Arachis hypogea agglutinin (FITC-PNA). Freezing was carried out in a programmable semen freezer, using 0.25 ml straws, in a total of 20 x 106 viable sperm per straw with glycerol as a cryoprotectant in a final concentration of 0.58 M. It was observed that, on average, a total of 7.25 ml of semen was collected from each bull. The viability and vitality rates were respectively 83.22 ± 7.52% and 3.8 ± 0.4 before freezing, decreasing to 58.81 ± 11.99% and 3.6 ± 0.6, respectively, after thawing. Regarding cytoplasmic droplets, it was observed that a high percentage of spermatozoa had medial cytoplasmic droplets (38.47%), with only 3.32% and 0.15% presenting proximal and distal cytoplasmic drops, respectively. By flow cytometry, it was observed that before freezing, the percentage of sperm with the damaged plasma membrane and intact acrosome was 3.61 ± 0.99%, increasing slightly to 4.21 ± 1.86% after cryopreservation (p<0.05). Regarding spermatozoa with damaged plasma membrane and acrosome, the percentage before freezing was 3.37±1.87%, increasing to 4.34 ±1.16% after thawing, and no significant differences were observed between these two values. For the percentage of sperm with the intact plasma membrane and damaged acrosome, this value was 2.04 ± 2.34% before freezing, decreasing to 0.89 ± 0.48% after thawing (p<0.05). The percentage of sperm with the intact plasma membrane and acrosome before freezing was 90.99±2.75%, with a slight decrease to 90.57±3.15% after thawing (p<0.05). From this study, it can be clearly concluded that, after the slaughtering of bulls, the spermatozoa can be recovered from the epididymis and cryopreserved, maintaining an excellent rate of sperm viability and quality after thawing.

Keywords: bovine semen, epididymis, cryopreservation, fertility assessment

Procedia PDF Downloads 87

Authors: Marinna Madrid, Nabiha Saklayen, Marinus Huber, Nicolas Vogel, Christos Boutopoulos, Michel Meunier, Eric Mazur

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Delivering material into cells is important for a diverse range of biological applications, including gene therapy, cellular engineering and imaging. We present a plasmonic substrate for delivering membrane-impermeable material into cells at high throughput and high efficiency while maintaining cell viability. The substrate fabrication is based on an affordable and fast colloidal self-assembly process. When illuminated with a femtosecond laser, the light interacts with the electrons at the surface of the metal substrate, creating localized surface plasmons that form bubbles via energy dissipation in the surrounding medium. These bubbles come into close contact with the cell membrane to form transient pores and enable entry of membrane-impermeable material via diffusion. We use fluorescence microscopy and flow cytometry to verify delivery of membrane-impermeable material into HeLa CCL-2 cells. We show delivery efficiency and cell viability data for a range of membrane-impermeable cargo, including dyes and biologically relevant material such as siRNA. We estimate the effective pore size by determining delivery efficiency for hard fluorescent spheres with diameters ranging from 20 nm to 2 um. To provide insight to the cell poration mechanism, we relate the poration data to pump-probe measurements of micro- and nano-bubble formation on the plasmonic substrate. Finally, we investigate substrate stability and reusability by using scanning electron microscopy (SEM) to inspect for damage on the substrate after laser treatment. SEM images show no visible damage. Our findings indicate that self-assembled plasmonic substrates are an affordable tool for high-throughput, high-efficiency delivery of material into mammalian cells.

Keywords: femtosecond laser, intracellular delivery, plasmonic, self-assembly

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Authors: Faozia A. S. Abuhtana, Yahia S. Abujnah, Said O. Gnann

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Ultra High Temperature (UHT) processed milk is widely distributed and preferred in numerous countries all over the world due its relatively high quality and long shelf life. Because of the notable high consumption rate of UHT in Libya in addition to negligible studies related to such product on the local level, this study was designed to assess the shelf life of locally produced as well as imported reconstituted sterilized whole milk samples marketed in Tripoli, Libya . Four locally produced vs. three imported brands were used in this study. All samples were stored at room temperature (25± 2C ) for 8 month long period, and subjected to physical, chemical, microbiological and sensory tests. These tests included : measurement of pH, specific gravity, percent acidity, and determination of fat, protein and melamine content. Microbiological tests included total aerobic count, total psychotropic bacteria, total spore forming bacteria and total coliform counts. Results indicated no detection of microbial growth of any type during the study period, in addition to no detection of melamine in all samples. On the other hand, a gradual decline in pH accompanied with gradual increase in % acidity of both locally produced and imported samples was observed. Such changes in both pH and % acidity reached their lowest and highest values respectively during the 24th week of storage. For instance pH values were (6.40, 6.55, 6.55, 6.15) and (6.30, 6.50, 6.20) for local and imported brands respectively. On the other hand, % acidity reached (0.185, 0181, 0170, 0183) and (0180, 0.180, 0.171) at the 24th week for local and imported brands respectively. Similar pattern of decline was also observed in specific gravity, fat and protein content in some local and imported samples especially at later stages of the study. In both cases, some of the recorded pH values, % acidity, sp. gravity and fat content were in violation of the accepted limits set by Libyan standard no. 356 for sterilized milk. Such changes in pH, % acidity and other UHT sterilized milk constituents during storage were coincided with a gradual decrease in the degree of acceptance of the stored milk samples of both types as shown by sensory scores recorded by the panelists. In either case degree of acceptance was significantly low at late stages of storage and most milk samples became relatively unacceptable after the 18th and 20th week for both untrained and trained panelists respectively.

Keywords: UHT milk, shelf life, quality, gravity, bacteria

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Authors: Supatchalee Sirichokworrakit, Pannin Intasen, Chansuda Angkawut

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Hom Nin rice (Oryza Sativa L.) was processed into flour and used to substitute wheat flour in donuts. The donuts were prepared with 0, 20, 40, 60, and 80% Hom Nin rice flour (HNF). The donuts were subjected to proximate, texture, color and sensory evaluations. The results of the study revealed that the ash, moisture, crude fiber contents increased while crude fat and protein contents decreased as the level of HNF increased. The hardness and chewiness of donut increased as the HNF increased but the cohesiveness, springiness, and specific volume decreased. Color of donut (L*, a*, and b* values) decreased with the addition of HNF. Overall acceptability for the 20-40% HNF additions did not differ significantly from the score of the 100% wheat flour.

Keywords: Hom Nin rice, donut, texture evaluation, sensory evaluation

Procedia PDF Downloads 293