Search results for: recombinant protein expression
Commenced in January 2007
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Paper Count: 3830

Search results for: recombinant protein expression

170 Computational Approaches to Study Lineage Plasticity in Human Pancreatic Ductal Adenocarcinoma

Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles

Abstract:

Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.

Keywords: deconvolution, imaging, microenvironment, PDAC

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169 Valorisation of Food Waste Residue into Sustainable Bioproducts

Authors: Krishmali N. Ekanayake, Brendan J. Holland, Colin J. Barrow, Rick Wood

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Globally, more than one-third of all food produced is lost or wasted, equating to 1.3 billion tonnes per year. Around 31.2 million tonnes of food waste are generated across the production, supply, and consumption chain in Australia. Generally, the food waste management processes adopt environmental-friendly and more sustainable approaches such as composting, anerobic digestion and energy implemented technologies. However, unavoidable, and non-recyclable food waste ends up as landfilling and incineration that involve many undesirable impacts and challenges on the environment. A biorefinery approach contributes to a waste-minimising circular economy by converting food and other organic biomass waste into valuable outputs, including feeds, nutrition, fertilisers, and biomaterials. As a solution, Green Eco Technologies has developed a food waste treatment process using WasteMaster system. The system uses charged oxygen and moderate temperatures to convert food waste, without bacteria, additives, or water, into a virtually odour-free, much reduced quantity of reusable residual material. In the context of a biorefinery, the WasteMaster dries and mills food waste into a form suitable for storage or downstream extraction/separation/concentration to create products. The focus of the study is to determine the nutritional composition of WasteMaster processed residue to potential develop aquafeed ingredients. The global aquafeed industry is projected to reach a high value market in future, which has shown high demand for the aquafeed products. Therefore, food waste can be utilized for aquaculture feed development by reducing landfill. This framework will lessen the requirement of raw crops cultivation for aquafeed development and reduce the aquaculture footprint. In the present study, the nutritional elements of processed residue are consistent with the input food waste type, which has shown that the WasteMaster is not affecting the expected nutritional distribution. The macronutrient retention values of protein, lipid, and nitrogen free extract (NFE) are detected >85%, >80%, and >95% respectively. The sensitive food components including omega 3 and omega 6 fatty acids, amino acids, and phenolic compounds have been found intact in each residue material. Preliminary analysis suggests a price comparability with current aquafeed ingredient cost making the economic feasibility. The results suggest high potentiality of aquafeed development as 5 to 10% of the ingredients to replace/partially substitute other less sustainable ingredients across biorefinery setting. Our aim is to improve the sustainability of aquaculture and reduce the environmental impacts of food waste.

Keywords: biorefinery, ffood waste residue, input, wasteMaster

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168 Upgrade of Value Chains and the Effect on Resilience of Russia’s Coal Industry and Receiving Regions on the Path of Energy Transition

Authors: Sergey Nikitenko, Vladimir Klishin, Yury Malakhov, Elena Goosen

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Transition to renewable energy sources (solar, wind, bioenergy, etc.) and launching of alternative energy generation has weakened the role of coal as a source of energy. The Paris Agreement and assumption of obligations by many nations to orderly reduce CO₂ emissions by means of technological modernization and climate change adaptation has abridged coal demand yet more. This paper aims to assess current resilience of the coal industry to stress and to define prospects for coal production optimization using high technologies pursuant to global challenges and requirements of energy transition. Our research is based on the resilience concept adapted to the coal industry. It is proposed to divide the coal sector into segments depending on the prevailing value chains (VC). Four representative models of VC are identified in the coal sector. The most promising lines of upgrading VC in the coal industry include: •Elongation of VC owing to introduction of clean technologies of coal conversion and utilization; •Creation of parallel VC by means of waste management; •Branching of VC (conversion of a company’s VC into a production network). The upgrade effectiveness is governed in many ways by applicability of advanced coal processing technologies, usability of waste, expandability of production, entrance to non-rival markets and localization of new segments of VC in receiving regions. It is also important that upgrade of VC by means of formation of agile high-tech inter-industry production networks within the framework of operating surface and underground mines can reduce social, economic and ecological risks associated with closure of coal mines. Such promising route of VC upgrade is application of methanotrophic bacteria to produce protein to be used as feed-stuff in fish, poultry and cattle breeding, or in production of ferments, lipoids, sterols, antioxidants, pigments and polysaccharides. Closed mines can use recovered methane as a clean energy source. There exist methods of methane utilization from uncontrollable sources, including preliminary treatment and recovery of methane from air-and-methane mixture, or decomposition of methane to hydrogen and acetylene. Separated hydrogen is used in hydrogen fuel cells to generate power to feed the process of methane utilization and to supply external consumers. Despite the recent paradigm of carbon-free energy generation, it is possible to preserve the coal mining industry using the differentiated approach to upgrade of value chains based on flexible technologies with regard to specificity of mining companies.

Keywords: resilience, resilience concept, resilience indicator, resilience in the Russian coal industry, value chains

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167 The Rise and Effects of Social Movement on Ethnic Relations in Malaysia: The Bersih Movement as a Case Study

Authors: Nur Rafeeda Daut

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The significance of this paper is to provide an insight on the role of social movement in building stronger ethnic relations in Malaysia. In particular, it focuses on how the BERSIH movement have been able to bring together the different ethnic groups in Malaysia to resist the present political administration that is seen to manipulate the electoral process and oppress the basic freedom of expression of Malaysians. Attention is given on how and why this group emerged and its mobilisation strategies. Malaysia which is a multi-ethnic and multi-religious society gained its independence from the British in 1957. Like many other new nations, it faces the challenges of nation building and governance. From economic issues to racial and religious tension, Malaysia is experiencing high level of corruption and income disparity among the different ethnic groups. The political parties in Malaysia are also divided along ethnic lines. BERSIH which is translated as ‘clean’ is a movement which seeks to reform the current electoral system in Malaysia to ensure equality, justice, free and fair elections. It was originally formed in 2007 as a joint committee that comprised leaders from political parties, civil society groups and NGOs. In April 2010, the coalition developed as an entirely civil society movement unaffiliated to any political party. BERSIH claimed that the electoral roll in Malaysia has been marred by fraud and other irregularities. In 2015, the BERSIH movement organised its biggest rally in Malaysia which also includes 38 other rallies held internationally. Supporters of BERSIH that participated in the demonstration were comprised of all the different ethnic groups in Malaysia. In this paper, two social movement theories are used: resource mobilization theory and political opportunity structure to explain the emergence and mobilization of the BERSIH movement in Malaysia. Based on these two theories, corruption which is believed to have contributed to the income disparity among Malaysians has generated the development of this movement. The rise of re-islamisation values propagated by certain groups in Malaysia and the shift in political leadership has also created political opportunities for this movement to emerge. In line with the political opportunity structure theory, the BERSIH movement will continue to create more opportunities for the empowerment of civil society and the unity of ethnic relations in Malaysia. Comparison is made on the degree of ethnic unity in the country before and after BERSIH was formed. This would include analysing the level of re-islamisation values and also the level of corruption in relation to economic income under the premiership of the former Prime Minister Mahathir and the present Prime Minister Najib Razak. The country has never seen such uprisings like BERSIH where ethnic groups which over the years have been divided by ethnic based political parties and economic disparity joined together with a common goal for equality and fair elections. As such, the BERSIH movement is a unique case where it illustrates the change of political landscape, ethnic relations and civil society in Malaysia.

Keywords: ethnic relations, Malaysia, political opportunity structure, resource mobilization theory and social movement

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166 A High Amylose-Content and High-Yielding Elite Line Is Favorable to Cook 'Nanhan' (Semi-Soft Rice) for Nursing Care Food Particularly for Serving Aged Persons

Authors: M. Kamimukai, M. Bhattarai, B. B. Rana, K. Maeda, H. B. Kc, T. Kawano, M. Murai

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Most of the aged people older than 70 have difficulty in chewing and swallowing more or less. According to magnitude of this difficulty, gruel, “nanhan” (semi-soft rice) and ordinary cooked rice are served in general, particularly in sanatoriums and homes for old people in Japan. Nanhan is the name of a cooked rice used in Japan, having softness intermediate between gruel and ordinary cooked rice, which is boiled with intermediate amount of water between those of the latter two kinds of cooked rice. In the present study, nanhan was made in the rate of 240g of water to 100g of milled rice with an electric rice cooker. Murai developed a high amylose-content and high-yielding elite line ‘Murai 79’. Sensory eating-quality test was performed for nanhan and ordinary cooked rice of Murai 79 and the standard variety ‘Hinohikari’ which is a high eating-quality variety representative in southern Japan. Panelists (6 to 14 persons) scored each cooked rice in six items viz. taste, stickiness, hardness, flavor, external appearance and overall evaluation. Grading (-3 ~ +3) in each trait was performed, regarding the value of the standard variety Hinohikari as 0. Paddy rice produced in a farmer’s field in 2013 and 2014 and in an experimental field of Kochi University in 2015 and 2016 were used for the sensory test. According to results of the sensory eating-quality test for nanhan, Murai 79 is higher in overall evaluation than Hinohikari in the four years. The former was less sticky than the latter in the four years, but the former was statistically significantly harder than the latter throughout the four years. In external appearance, the former was significantly higher than the latter in the four years. In the taste, the former was significantly higher than the latter in 2014, but significant difference was not noticed between them in the other three years. There were no significant differences throughout the four years in flavor. Regarding amylose content, Murai 79 is higher by 3.7 and 5.7% than Hinohikari in 2015 and 2016, respectively. As for protein content, Murai 79 was higher than Hinohikari in 2015, but the former was lower than the latter in 2016. Consequently, the nanhan of Murai 79 was harder and less sticky, keeping the shape of grains as compared with that of Hinohikari, which may be due to its higher amylose content. Hence, the nanhan of Murai 79 may be recognized as grains more easily in a human mouth, which could make easier the continuous performance of mastication and deglutition particularly in aged persons. Regarding ordinary cooked rice, Murai 79 was similar to or higher in both overall evaluation and external appearance as compared with Hinohikari, despite its higher hardness and lower stickiness. Additionally, Murai 79 had brown-rice yield of 1.55 times as compared with Hinohikari, suggesting that it would enable to supply inexpensive rice for making nanhan with high quality particularly for aged people in Japan.

Keywords: high-amylose content, high-yielding rice line, nanhan, nursing care food, sensory eating quality test

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165 The Structural Alteration of DNA Native Structure of Staphylococcus aureus Bacteria by Designed Quinoxaline Small Molecules Result in Their Antibacterial Properties

Authors: Jeet Chakraborty, Sanjay Dutta

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Antibiotic resistance by bacteria has proved to be a severe threat to mankind in recent times, and this fortifies an urgency to design and develop potent antibacterial small molecules/compounds with nonconventional mechanisms than the conventional ones. DNA carries the genetic signature of any organism, and bacteria maintain their genomic DNA inside the cell in a well-regulated compact form with the help of various nucleoid associated proteins like HU, HNS, etc. These proteins control various fundamental processes like gene expression, replication, etc., inside the cell. Alteration of the native DNA structure of bacteria can lead to severe consequences in cellular processes inside the bacterial cell that ultimately result in the death of the organism. The change in the global DNA structure by small molecules initiates a plethora of cellular responses that have not been very well investigated. Echinomycin and Triostin-A are biologically active Quinoxaline small molecules that typically consist of a quinoxaline chromophore attached with an octadepsipeptide ring. They bind to double-stranded DNA in a sequence-specific way and have high activity against a wide variety of bacteria, mainly against Gram-positive ones. To date, few synthetic quinoxaline scaffolds were synthesized, displaying antibacterial potential against a broad scale of pathogenic bacteria. QNOs (Quinoxaline N-oxides) are known to target DNA and instigate reactive oxygen species (ROS) production in bacteria, thereby exhibiting antibacterial properties. The divergent role of Quinoxaline small molecules in medicinal research qualifies them for the evaluation of their antimicrobial properties as a potential candidate. The previous study from our lab has given new insights on a 6-nitroquinoxaline derivative 1d as an intercalator of DNA, which induces conformational changes in DNA upon binding.7 The binding event observed was dependent on the presence of a crucial benzyl substituent on the quinoxaline moiety. This was associated with a large induced CD (ICD) appearing in a sigmoidal pattern upon the interaction of 1d with dsDNA. The induction of DNA superstructures by 1d at high Drug:DNA ratios was observed that ultimately led to DNA condensation. Eviction of invitro-assembled nucleosome upon treatment with a high dose of 1d was also observed. In this work, monoquinoxaline derivatives of 1d were synthesized by various modifications of the 1d scaffold. The set of synthesized 6-nitroquinoxaline derivatives along with 1d were all subjected to antibacterial evaluation across five different bacteria species. Among the compound set, 3a displayed potent antibacterial activity against Staphylococcus aureus bacteria. 3a was further subjected to various biophysical studies to check whether the DNA structural alteration potential was still intact. The biological response of S. aureus cells upon treatment with 3a was studied using various cell biology processes, which led to the conclusion that 3d can initiate DNA damage in the S. aureus cells. Finally, the potential of 3a in disrupting preformed S.aureus and S.epidermidis biofilms was also studied.

Keywords: DNA structural change, antibacterial, intercalator, DNA superstructures, biofilms

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164 Genome-Wide Homozygosity Analysis of the Longevous Phenotype in the Amish Population

Authors: Sandra Smieszek, Jonathan Haines

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Introduction: Numerous research efforts have focused on searching for ‘longevity genes’. However, attempting to decipher the genetic component of the longevous phenotype have resulted in limited success and the mechanisms governing longevity remain to be explained. We conducted a genome-wide homozygosity analysis (GWHA) of the founder population of the Amish community in central Ohio. While genome-wide association studies using unrelated individuals have revealed many interesting longevity associated variants, these variants are typically of small effect and cannot explain the observed patterns of heritability for this complex trait. The Amish provide a large cohort of extended kinships allowing for in depth analysis via family-based approach excellent population due to its. Heritability of longevity increases with age with significant genetic contribution being seen in individuals living beyond 60 years of age. In our present analysis we show that the heritability of longevity is estimated to be increasing with age particularly on the paternal side. Methods: The present analysis integrated both phenotypic and genotypic data and led to the discovery of a series of variants, distinct for stratified populations across ages and distinct for paternal and maternal cohorts. Specifically 5437 subjects were analyzed and a subset of 893 successfully genotyped individuals was used to assess CHIP heritability. We have conducted the homozygosity analysis to examine if homozygosity is associated with increased risk of living beyond 90. We analyzed AMISH cohort genotyped for 614,957 SNPs. Results: We delineated 10 significant regions of homozygosity (ROH) specific for the age group of interest (>90). Of particular interest was ROH on chromosome 13, P < 0.0001. The lead SNPs rs7318486 and rs9645914 point to COL4A2 and our lead SNP. COL25A1 encodes one of the six subunits of type IV collagen, the C-terminal portion of the protein, known as canstatin, is an inhibitor of angiogenesis and tumor growth. COL4A2 mutations have been reported with a broader spectrum of cerebrovascular, renal, ophthalmological, cardiac, and muscular abnormalities. The second region of interest points to IRS2. Furthermore we built a classifier using the obtained SNPs from the significant ROH region with 0.945 AUC giving ability to discriminate between those living beyond to 90 years of age and beyond. Conclusion: In conclusion our results suggest that a history of longevity does indeed contribute to increasing the odds of individual longevity. Preliminary results are consistent with conjecture that heritability of longevity is substantial when we start looking at oldest fifth and smaller percentiles of survival specifically in males. We will validate all the candidate variants in independent cohorts of centenarians, to test whether they are robustly associated with human longevity. The identified regions of interest via ROH analysis could be of profound importance for the understanding of genetic underpinnings of longevity.

Keywords: regions of homozygosity, longevity, SNP, Amish

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163 Detection of Egg Proteins in Food Matrices (2011-2021)

Authors: Daniela Manila Bianchi, Samantha Lupi, Elisa Barcucci, Sandra Fragassi, Clara Tramuta, Lucia Decastelli

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Introduction: The undeclared allergens detection in food products plays a fundamental role in the safety of the allergic consumer. The protection of allergic consumers is guaranteed, in Europe, by Regulation (EU) No 1169/2011 of the European Parliament, which governs the consumer's right to information and identifies 14 food allergens to be mandatorily indicated on food labels: among these, an egg is included. An egg can be present as an ingredient or as contamination in raw and cooked products. The main allergen egg proteins are ovomucoid, ovalbumin, lysozyme, and ovotransferrin. This study presents the results of a survey conducted in Northern Italy aimed at detecting the presence of undeclared egg proteins in food matrices in the latest ten years (2011-2021). Method: In the period January 2011 - October 2021, a total of 1205 different types of food matrices (ready-to-eat, meats, and meat products, bakery and pastry products, baby foods, food supplements, pasta, fish and fish products, preparations for soups and broths) were delivered to Food Control Laboratory of Istituto Zooprofilattico Sperimentale of Piemonte Liguria and Valle d’Aosta to be analyzed as official samples in the frame of Regional Monitoring Plan of Food Safety or in the contest of food poisoning. The laboratory is ISO 17025 accredited, and since 2019, it has represented the National Reference Centre for the detection in foods of substances causing food allergies or intolerances (CreNaRiA). All samples were stored in the laboratory according to food business operator instructions and analyzed within the expiry date for the detection of undeclared egg proteins. Analyses were performed with RIDASCREEN®FAST Ei/Egg (R-Biopharm ® Italia srl) kit: the method was internally validated and accredited with a Limit of Detection (LOD) equal to 2 ppm (mg/Kg). It is a sandwich enzyme immunoassay for the quantitative analysis of whole egg powder in foods. Results: The results obtained through this study showed that egg proteins were found in 2% (n. 28) of food matrices, including meats and meat products (n. 16), fish and fish products (n. 4), bakery and pastry products (n. 4), pasta (n. 2), preparations for soups and broths (n.1) and ready-to-eat (n. 1). In particular, in 2011 egg proteins were detected in 5% of samples, in 2012 in 4%, in 2013, 2016 and 2018 in 2%, in 2014, 2015 and 2019 in 3%. No egg protein traces were detected in 2017, 2020, and 2021. Discussion: Food allergies occur in the Western World in 2% of adults and up to 8% of children. Allergy to eggs is one of the most common food allergies in the pediatrics context. The percentage of positivity obtained from this study is, however, low. The trend over the ten years has been slightly variable, with comparable data.

Keywords: allergens, food, egg proteins, immunoassay

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162 Effect of Different By-Products on Growth Performance, Carcass Characteristics and Serum Parameters of Growing Simmental Crossbred Cattle

Authors: Fei Wang, Jie Meng, Qingxiang Meng

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China is rich in straw and by-product resources, whose utilization has always been a hot topic. The objective of this study was to investigate the effect of feeding soybean straw and wine distiller’s grain as a replacement for corn stover on performance of beef cattle. Sixty Simmental×local crossbred bulls averaging 12 months old and 335.7 ± 39.1 kg of body weight (BW) were randomly assigned into four groups (15 animals per group) and allocated to a diet with 40% maize stover (MSD), a diet with 40% wrapping package maize silage (PMSD), a diet with 12% soybean straw plus 28% maize stover (SSD) and a diet with 12% wine distiller’s grain plus 28% maize stover (WDD). Bulls were fed ad libitum an TMR consisting of 36.0% maize, 12.5% of DDGS, 5.0% of cottonseed meal, 4.0% of soybean meal and 40.0% of by-product as described above. Treatment period lasted for 22 weeks, consisting of 1 week of dietary adaptation. The results showed that dry matter intake (DMI) was significantly higher (P < 0.01) for PMSD group than MSD and SSD groups during 0-7 week and 8-14week, and PMSD and WDD groups had higher (P < 0.05) DMI values than MSD and SSD groups during the whole period. Average daily gain (ADG) values were 1.56, 1.72, 1.68 and 1.58 kg for MSD, PMSD, SSD and WDD groups respectively, although the differences were not significant (P > 0.05). The value of blood sugar concentration was significantly higher (P < 0.01) for MSD group than WDD group, and the blood urea nitrogen concentration of SSD group was lower (P < 0.05) than MSD and WDD groups. No significant difference (P > 0.05) of serum total cholesterol, triglycerides or total protein content was observed among the different groups. Ten bulls with similar body weight were selected at the end of feeding trial and slaughtered for measurement of slaughtering performance, carcass quality and meat chemical composition. SSD group had significantly lower (P < 0.05) shear force value and cooking loss than MSD and PMSD groups. The pH values of MSD and SSD groups were lower (P < 0.05) than PMSD and WDD groups. WDD group had a higher fat color brightness (L*) value than PMSD and SSD groups. There were no significant differences in dressing percentage, meat percentage, top grade meat weight, ribeye area, marbling score, meat color and meat chemical compositions among different dietary treatments. Based on these results, the packed maize stover silage showed a potential of improving the average daily gain and feed intake of beef cattle. Soybean straw had a significant effect on improving the tenderness and reducing cooking loss of beef. In general, soybean straw and packed maize stover silage would be beneficial to nitrogen deposition and showed a potential to substitute maize stover in beef cattle diets.

Keywords: beef cattle, by-products, carcass quality, growth performance

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161 Anti-Angiogenic and Anti-Metastatic Effect of Aqueous Fraction from Euchelus Asper Methanolic Extract

Authors: Sweta Agrawal, Sachin Chaugule, Gargi Rane, Shashank More, Madhavi Indap

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Angiogenesis and metastasis are two of the most important hallmarks of cancer. Hence, most of the cancer therapies nowadays are multi-targeted so as to reduce resistance and have better efficacy. As synthetic molecules arise with a burden of their toxicities and side-effects, more and more research is being focussed on exploiting the vast natural resources of drugs, in the form of plants and animals. Although, the idea of using marine organisms as a source of pharmaceuticals is not new, the pace at which marine drugs are being discovered, has definitely up surged! In the present study, we have assessed the anti-angiogenic and in vitro anti-metastatic activity of aqueous fraction from the extract of marine gastropod Euchelus asper. The soft body of Euchelus Asper was extracted with methanol and named EAME. Partition chromatography of EAME gave three fractions EAME I, II and III. Biochemical analysis revealed the presence of proteins in EAME III. Preliminary analysis had revealed the anti-angiogenic activity was exhibited by EAME III out of the three fractions. Hereafter, EAME III (concentration 25µg/ml-400µg/ml) was tested on chick chorioallantoic membrane (CAM) model for the detailed analysis of its potential anti-angiogenic effect. In vitro testing of the fraction (concentration 0.25µg/ml - 1µg/ml), involved cytotoxicity by SRB assay, cell cycle analysis by flow cytometry and anti-proliferative effect by scratch wound healing assay on A549 lung carcinoma cells. Apart from this, a portion of treated CAM as well as conditioned medium from treated A549 were subjected to gelatin zymography for assessment of matrix metalloproteinases MMP-2 and MMP-9 levels. Our results revealed that EAME III exhibited significant anti-angiogenic activity on CAM which was also supported by histological observations. During histological studies of CAM, it was found that EAME III caused reduction in angiogenesis by altering the extracellular matrix of the CAM membrane. In vitro analysis disclosed that EAME III exhibited moderate cytotoxic effect on A549 cells and its effect was not dose-dependent. The results of flow cytometry confirmed that EAME III caused cell cycle arrest in A549 cell line as almost all of the treated cells were found in G1 phase. Further, the migration and proliferation of A549 was significantly reduced by EAME III as observed from the scratch wound assay. Moreover, Gelatin zymography analysis revealed that EAME III caused suppression of MMP-2 in CAM membrane and reduced MMP-9 and MMP-2 expression in A549 cells. This verified that the anti-angiogenic and anti-metastatic effects of EAME III were correlated with the suppression of MMP-2 and -9. To conclude, EAME III shows dual anti-tumour action by reducing angiogenesis and exerting anti-metastatic effect on lung cancer cells, thus it has the potential to be used as an anti-cancer agent against lung carcinoma.

Keywords: angiogenesis, anti-cancer, marine drugs, matrix metalloproteinases

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160 The ‘Othered’ Body: Deafness and Disability in Nina Raine’s Tribes

Authors: Nurten Çelik

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Under the new developments in science, medicine, sociology, psychology and literary theories, body studies has gained huge importance and the body has become a debatable issue. There has emerged, among sociologists and literary theorists, an overwhelming consensus that body is socially, politically and culturally perceived and constructed and thus, the position of an individual in the society is determined in accordance with his/her body image. In this regard, the most complicated point is the theoretical views propounded upon disability studies, where the disabled body is considered to be a site upon which social and political restrictions as well as repressions are inscribed. There has been the widely-accepted view that no matter what kind of disability it is, those with physical, mental or learning impairments face varied social, political and environmental obstacles that prevent them from being an active citizen, worker, lover and even a family member. In parallel with these approaches, the matter of the sufferings of disabled individuals attains its place in cinema and literature as well as in theatre studies under the category of disability theatre. One of the prominent plays that deal with physical disability came from the contemporary British playwright Nina Raine. In her awarded play Tribes, which premiered at the Royal Court Theatre in 2010, Raine develops the social strata where her deaf protagonist, Billy, caught up between two tribes – namely his family and his lover Slyvia, a member of the deaf community– experiences personal and social hardships due to his hearing impairment. In the play, intransigent and self-opinionated family members foster no sense of empathy towards Billy, there are noisy talking and shouting, but no communication, love, compassion or mutual understanding, and language becomes just a tool for the expression of rage and oppression. In the disordered atmosphere of the family life, Billy experiences isolation and loneliness. Billy’s hopes for success and love are destroyed when Slyvia, troubled between hearing and deafness, rejects him because she does not utterly grasp what Billy is experiencing. Drawing upon the hardships, Billy undergoes in his relationships with his family and his girlfriend, Tribes problematizes the concept of deafness and explores to what extent a deaf person can find a place in the hearing world. Setting ‘the disabled’ bodies against ‘the abled’ bodies in a family, a microcosm of the society where bodies are socially shaped and constructed, Tribes dramatizes how the disabled bodies are disenfranchised, stigmatised, marginalized and othered on the grounds that they are socially misfit. Tribes, with a specific focus on the dysfunctional family, shows that the lack of communication and empathy numbs the characters to the feelings of each other and thereby, they become more disabled than Billy. In conclusion, this paper, with the reference to the embodiment of disability and social theories, aims to explore how disabled bodies are socially marked and segregated from family and society.

Keywords: body, deafness, disability, disability theatre, Nina Raine, tribes

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159 Functional Analysis of Variants Implicated in Hearing Loss in a Cohort from Argentina: From Molecular Diagnosis to Pre-Clinical Research

Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

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Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

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158 Bioresorbable Medicament-Eluting Grommet Tube for Otitis Media with Effusion

Authors: Chee Wee Gan, Anthony Herr Cheun Ng, Yee Shan Wong, Subbu Venkatraman, Lynne Hsueh Yee Lim

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Otitis media with effusion (OME) is the leading cause of hearing loss in children worldwide. Surgery to insert grommet tube into the eardrum is usually indicated for OME unresponsive to antimicrobial therapy. It is the most common surgery for children. However, current commercially available grommet tubes are non-bioresorbable, not drug-treated, with unpredictable duration of retention on the eardrum to ventilate middle ear. Their functionality is impaired when clogged or chronically infected, requiring additional surgery to remove/reinsert grommet tubes. We envisaged that a novel fully bioresorbable grommet tube with sustained antibiotic release technology could address these drawbacks. In this study, drug-loaded bioresorbable poly(L-lactide-co-ε-caprolactone)(PLC) copolymer grommet tubes were fabricated by microinjection moulding technique. In vitro drug release and degradation model of PLC tubes were studied. Antibacterial property was evaluated by incubating PLC tubes with P. aeruginosa broth. Surface morphology was analyzed using scanning electron microscopy. A preliminary animal study was conducted using guinea pigs as an in vivo model to evaluate PLC tubes with and without drug, with commercial Mini Shah grommet tube as comparison. Our in vitro data showed sustained drug release over 3 months. All PLC tubes revealed exponential degradation profiles over time. Modeling predicted loss of tube functionality in water to be approximately 14 weeks and 17 weeks for PLC with and without drug, respectively. Generally, PLC tubes had less bacteria adherence, which were attributed to the much smoother tube surfaces compared to Mini Shah. Antibiotic from PLC tube further made bacteria adherence on surface negligible. They showed neither inflammation nor otorrhea after 18 weeks post-insertion in the eardrums of guinea pigs, but had demonstrated severe degree of bioresorption. Histology confirmed the new PLC tubes were biocompatible. Analyses on the PLC tubes in the eardrums showed bioresorption profiles close to our in vitro degradation models. The bioresorbable antibiotic-loaded grommet tubes showed good predictability in functionality. The smooth surface and sustained release technology reduced the risk of tube infection. Tube functional duration of 18 weeks allowed sufficient ventilation period to treat OME. Our ongoing studies include modifying the surface properties with protein coating, optimizing the drug dosage in the tubes to enhance their performances, evaluating their functional outcome on hearing after full resoption of grommet tube and healing of eardrums, and developing animal model with OME to further validate our in vitro models.

Keywords: bioresorbable polymer, drug release, grommet tube, guinea pigs, otitis media with effusion

Procedia PDF Downloads 432
157 Encephalon-An Implementation of a Handwritten Mathematical Expression Solver

Authors: Shreeyam, Ranjan Kumar Sah, Shivangi

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Recognizing and solving handwritten mathematical expressions can be a challenging task, particularly when certain characters are segmented and classified. This project proposes a solution that uses Convolutional Neural Network (CNN) and image processing techniques to accurately solve various types of equations, including arithmetic, quadratic, and trigonometric equations, as well as logical operations like logical AND, OR, NOT, NAND, XOR, and NOR. The proposed solution also provides a graphical solution, allowing users to visualize equations and their solutions. In addition to equation solving, the platform, called CNNCalc, offers a comprehensive learning experience for students. It provides educational content, a quiz platform, and a coding platform for practicing programming skills in different languages like C, Python, and Java. This all-in-one solution makes the learning process engaging and enjoyable for students. The proposed methodology includes horizontal compact projection analysis and survey for segmentation and binarization, as well as connected component analysis and integrated connected component analysis for character classification. The compact projection algorithm compresses the horizontal projections to remove noise and obtain a clearer image, contributing to the accuracy of character segmentation. Experimental results demonstrate the effectiveness of the proposed solution in solving a wide range of mathematical equations. CNNCalc provides a powerful and user-friendly platform for solving equations, learning, and practicing programming skills. With its comprehensive features and accurate results, CNNCalc is poised to revolutionize the way students learn and solve mathematical equations. The platform utilizes a custom-designed Convolutional Neural Network (CNN) with image processing techniques to accurately recognize and classify symbols within handwritten equations. The compact projection algorithm effectively removes noise from horizontal projections, leading to clearer images and improved character segmentation. Experimental results demonstrate the accuracy and effectiveness of the proposed solution in solving a wide range of equations, including arithmetic, quadratic, trigonometric, and logical operations. CNNCalc features a user-friendly interface with a graphical representation of equations being solved, making it an interactive and engaging learning experience for users. The platform also includes tutorials, testing capabilities, and programming features in languages such as C, Python, and Java. Users can track their progress and work towards improving their skills. CNNCalc is poised to revolutionize the way students learn and solve mathematical equations with its comprehensive features and accurate results.

Keywords: AL, ML, hand written equation solver, maths, computer, CNNCalc, convolutional neural networks

Procedia PDF Downloads 91
156 Bone Mineralization in Children with Wilson’s Disease

Authors: Shiamaa Eltantawy, Gihan Sobhy, Alif Alaam

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Wilson disease, or hepatolenticular degeneration, is an autosomal recessive disease that results in excess copper buildup in the body. It primarily affects the liver and basal ganglia of the brain, but it can affect other organ systems. Musculoskeletal abnormalities, including premature osteoarthritis, skeletal deformity, and pathological bone fractures, can occasionally be found in WD patients with a hepatic or neurologic type. The aim was to assess the prevalence of osteoporosis and osteopenia in Wilson’s disease patients. This case-control study was conducted on ninety children recruited from the inpatient ward and outpatient clinic of the Paediatric Hepatology, Gastroenterology, and Nutrition department of the National Liver Institute at Menofia University, aged from 1 to 18 years. Males were 49, and females were 41. Children were divided into three groups: (Group I) consisted of thirty patients with WD; (Group II) consisted of thirty patients with chronic liver disease other than WD; (Group III) consisted of thirty age- and sex-matched healthy The exclusion criteria were patients with hyperparathyroidism, hyperthyroidism, renal failure, Cushing's syndrome, and patients on certain drugs such as chemotherapy, anticonvulsants, or steroids. All patients were subjected to the following: 1- Full history-taking and clinical examination. 2-Laboratory investigations: (FBC,ALT,AST,serum albumin, total protein, total serum bilirubin,direct bilirubin,alkaline phosphatase, prothrombin time, serum critine,parathyroid hormone, serum calcium, serum phosphrus). 3-Bone mineral density (BMD, gm/cm2) values were measured by dual-energy X-ray absorptiometry (DEXA). The results revealed that there was a highly statistically significant difference between the three groups regarding the DEXA scan, and there was no statistically significant difference between groups I and II, but the WD group had the lowest bone mineral density. The WD group had a large number of cases of osteopenia and osteoporosis, but there was no statistically significant difference with the group II mean, while a high statistically significant difference was found when compared to group III. In the WD group, there were 20 patients with osteopenia, 4 patients with osteoporosis, and 6 patients who were normal. The percentages were 66.7%, 13.3%, and 20%, respectively. Therefore, the largest number of cases in the WD group had osteopenia. There was no statistically significant difference found between WD patients on different treatment regimens regarding DEXA scan results (Z-Score). There was no statistically significant difference found between patients in the WD group (normal, osteopenic, or osteoporotic) regarding phosphorus (mg/dL), but there was a highly statistically significant difference found between them regarding ionised Ca (mmol/L). Therefore, there was a decrease in bone mineral density when the Ca level was decreased. In summary, Wilson disease is associated with bone demineralization. The largest number of cases in the WD group in our study had osteopenia (66.7%). Different treatment regimens (zinc monotherapy, Artamin, and zinc) as well as different laboratory parameters have no effect on bone mineralization in WD cases. Decreased ionised Ca is associated with low BMD in WD patients. Children with WD should be investigated for BMD.

Keywords: wilson disease, Bone mineral density, liver disease, osteoporosis

Procedia PDF Downloads 39
155 Potential of Water Purification of Turbid Surface Water Sources in Remote Arid and Semi-Arid Rural Areas of Rajasthan by Moringa Oleifera (Drumstick) Tree Seeds

Authors: Pomila Sharma

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Rajasthan is among regions with greatest climate sensitivity and lowest adaptive capabilities. In many parts of the Rajasthan surface water which can be highly turbid and contaminated with fecal coliform bacteria is used for drinking purposes. The majority rely almost exclusively upon traditional sources of highly turbid and untreated pathogenic surface water for their domestic water needs. In many parts of rural areas of Rajasthan, it is still difficult to obtain clean water, especially remote habitations with no groundwater due to quality issues or depletion and limited feasibility to connect with surface water schemes due to low density of population in these areas to justify large infrastructure investment. The most viable sources are rain water harvesting, community managed open wells, private wells, ponds and small-scale irrigation reservoirs have often been the main traditional sources of rural drinking water. Turbidity is conventionally removed by treating the water with expensive chemicals. This study has to investigate the use of crushed seeds from the tree Moringa oleifera (drumstick) as a natural alternative to conventional coagulant chemicals. The use of Moringa oleifera seed powder can produce potable water of higher quality than the original source. Moringa oleifera a native species of northern India, the tree is now grown extensively throughout the tropics and found in many countries of Africa, Asia & South America. The seeds of tree contains significant quantities of low molecular weight, water soluble proteins which carries the positive charge when the crushed seeds are added to water. This protein binds in raw water with negatively charged turbid water with bacteria, clay, algae, etc. Under proper mixing, these particles make flocks, which may be left to settle by gravity or be removed by filtration. Using Moringa oleifera as a replacement coagulation in such surface sources of arid and semi-arid areas can meet the need for water purification in remote places of Rajasthan state of India. The present study accesses to find out laboratory based investigation of the effect of seeds of Moringa tree on its coagulation effectiveness (purification) using turbid water samples of surface source of the Rajasthan state. In this study, moringa seed powder showed that filtering with seed powder may diminish water pollution and bacterial counts. Results showed Moringa oleifera seeds coagulate 90-95% of turbidity and color efficiently leading to an aesthetically clear supernatant & reduced about 85-90% of bacterial load reduction in samples.

Keywords: bacterial load, coagulant, turbidity, water purification

Procedia PDF Downloads 120
154 Sexual Behaviour and Psychological Well-Being of a Group of African Adolescent Males in Alice, Eastern Cape

Authors: Jabulani Gilford Kheswa, Thembelihle Lobi

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From a cultural perspective, expression of hegemonic masculinity in South Africa continues to escalate among adolescent males who grow up in communities lacking in role models and recreational facilities. However, when the schools are constructive, and peer influence is positive, adolescent male can potentially express character strengths and lead a meaningful life. Drawing from Bronfenbrenner’s Ecological Model and Keyes and Ryff’s six dimensions of psychological well-being and mental health, such youth may exemplify positive self-esteem, problem- focused coping strategies, condom self-efficacy, good leadership skills, enhanced motivation and a positive emotional state, which buffer against risky sexual behaviors. This paper was aimed at investigating the relationships between adolescent males’ sexual behavior and psychological well-being. This study employed a quantitative research to collect data from 54 Xhosa-speaking adolescent males from one school high school in Fort Beaufort, Eastern Cape, South Africa. These learners were from grade nine, ten and eleven with their ages ranging from 14 to 20. Prior the research commenced, the school principal and caregivers of the learners who participated in the study, gave their informed consent. Self- administered closed-ended questionnaire with Section A (that is, biographical information) and Section B with each question rated on the 5–point Likert scale was used. The advantages of questionnaires include a high response rate as they require less time and offer anonymity because participants’ names are not identified. The SPSS version 18 was used for statistical data analysis. The mean age was 16.83 with a standard deviation of 1.611. 44.4% of the participants were from grade 9, 33.3% from grade 10 and 22.2% from grade 11. The Chronbach alpha of 0.79 was yielded, with respect to self- esteem of adolescent males. In this study, 76.9% reported to attend church services whilst 23% indicated not to attend church services. A further 96.2% of adolescent males indicated to have good relations with guardians while only 3.8% had poorer relations. A large proportion of adolescent males (72.9%) indicated to high-quality friendship as opposed to 27.1% who reported being receiving negative guidance from peers. Other findings revealed that 81.1% of the participants’ parents do not drink alcohol, and they cope at school as 79.6% reported protective factors as attributable towards non-engagement to risky sexual practices. As a result, 81.4% of participants reported not to participate in criminal activities although 85% of the participants indicated that in their school there are drugs. It could be speculated from this study that adolescent males whose caregivers are authoritative, find purpose in life and are most likely to be socially and academically competent. This paper leads to further research interest into mental health, coping strategies and sexual decision-making skills of the youth in South Africa.

Keywords: church, mental health, school, sexual behaviour

Procedia PDF Downloads 232
153 Regulating the Ottomans on Turkish Television and the Making of Good Citizens

Authors: Chien Yang Erdem

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This paper takes up the proliferating historical dramas and children’s programs featuring the Ottoman-Islamic legacy on Turkish television as a locus where the processes of subjectification take place. A critical analysis of this emergent cultural phenomenon reveals an alliance of neoliberal and neoconservative political rationalities based on which the Turkish media is restructured to transform society. The existing debates have focused on how the Ottoman historical dramas manifest the Justice and Development Party’s (Adalet ve Kalkınma Partisi) neo-Ottomanist ideology and foreign policy. However, this approach tends to overlook the more complex relationship between the media, government, and society. Employing Michel Foucault’s notion of 'technologies of the self,' this paper aims to examine the governing practices that are deployed to regulate the media and to transform individual citizens into governable subjects in contemporary Turkey. First, through a brief discussion of recent development of the Turkish media towards an authoritarian model, the paper suggests that the relation between the Ottoman television drama and the political subject in question cannot be adequately examined without taking into account the force of the market. Second, by focusing on the managerial restructuring of the Turkish Television and Radio Corporation (Türkiye Radyo ve Televizyon Kurumu), the paper aims to illustrate the rationale and process through which the Turkish media sector is transformed into an integral part of the free market where the government becomes a key actor. The paper contends that this new sphere of free market is organized in a way that enables direct interference of the government and divides media practitioners and consumers into opposing categories through their own participation in the media market. On the one hand, a 'free subject' is constituted based on the premise that the market is a sphere where individuals are obliged to exercise their right to freedom (of choice, lifestyle, and expression). On the other hand, this 'free subject' is increasingly subjugated to such disciplinary practices as censorship for being on the wrong side of the government. Finally, the paper examines the relation between the restructured Turkish media market and the proliferation of Ottoman television drama in the 2010s. The study maintains that the reorganization of the media market has produced a condition where private sector is encouraged to take an active role in reviving Turkey’s Ottoman-Islamic cultural heritage and promulgating moral-religious values. Paying specific attention to the controversial case of Magnificent Century (Muhteşem Yüzyıl) in contrast with TRT’s Ottoman historical drama and children’s programs, the paper aims to identify the ways in which individual citizens are directed to conduct themselves as a virtuous citizenry. It is through the double movement between the governing practices associated with the media market and those concerning the making of a 'conservative generation' that a subject of citizenry of new Turkey is constituted.

Keywords: neoconservatism, neoliberalism, ottoman historical drama, technologies of the self, Turkish television

Procedia PDF Downloads 118
152 Food Safety in Wine: Removal of Ochratoxin a in Contaminated White Wine Using Commercial Fining Agents

Authors: Antònio Inês, Davide Silva, Filipa Carvalho, Luís Filipe-Riberiro, Fernando M. Nunes, Luís Abrunhosa, Fernanda Cosme

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The presence of mycotoxins in foodstuff is a matter of concern for food safety. Mycotoxins are toxic secondary metabolites produced by certain molds, being ochratoxin A (OTA) one of the most relevant. Wines can also be contaminated with these toxicants. Several authors have demonstrated the presence of mycotoxins in wine, especially ochratoxin A. Its chemical structure is a dihydro-isocoumarin connected at the 7-carboxy group to a molecule of L-β-phenylalanine via an amide bond. As these toxicants can never be completely removed from the food chain, many countries have defined levels in food in order to attend health concerns. OTA contamination of wines might be a risk to consumer health, thus requiring treatments to achieve acceptable standards for human consumption. The maximum acceptable level of OTA in wines is 2.0 μg/kg according to the Commission regulation No. 1881/2006. Therefore, the aim of this work was to reduce OTA to safer levels using different fining agents, as well as their impact on white wine physicochemical characteristics. To evaluate their efficiency, 11 commercial fining agents (mineral, synthetic, animal and vegetable proteins) were used to get new approaches on OTA removal from white wine. Trials (including a control without addition of a fining agent) were performed in white wine artificially supplemented with OTA (10 µg/L). OTA analyses were performed after wine fining. Wine was centrifuged at 4000 rpm for 10 min and 1 mL of the supernatant was collected and added of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v). Also, the solid fractions obtained after fining, were centrifuged (4000 rpm, 15 min), the resulting supernatant discarded, and the pellet extracted with 1 mL of the above solution and 1 mL of H2O. OTA analysis was performed by HPLC with fluorescence detection. The most effective fining agent in removing OTA (80%) from white wine was a commercial formulation that contains gelatin, bentonite and activated carbon. Removals between 10-30% were obtained with potassium caseinate, yeast cell walls and pea protein. With bentonites, carboxymethylcellulose, polyvinylpolypyrrolidone and chitosan no considerable OTA removal was verified. Following, the effectiveness of seven commercial activated carbons was also evaluated and compared with the commercial formulation that contains gelatin, bentonite and activated carbon. The different activated carbons were applied at the concentration recommended by the manufacturer in order to evaluate their efficiency in reducing OTA levels. Trial and OTA analysis were performed as explained previously. The results showed that in white wine all activated carbons except one reduced 100% of OTA. The commercial formulation that contains gelatin, bentonite and activated carbon reduced only 73% of OTA concentration. These results may provide useful information for winemakers, namely for the selection of the most appropriate oenological product for OTA removal, reducing wine toxicity and simultaneously enhancing food safety and wine quality.

Keywords: wine, ota removal, food safety, fining

Procedia PDF Downloads 514
151 Mycotoxin Bioavailability in Sparus Aurata Muscle After Human Digestion and Intestinal Transport (Caco-2/HT-29 Cells) Simulation

Authors: Cheila Pereira, Sara C. Cunha, Miguel A. Faria, José O. Fernandes

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The increasing world population brings several concerns, one of which is food security and sustainability. To meet this challenge, aquaculture, the farming of aquatic animals and plants, including fish, mollusks, bivalves, and algae, has experienced sustained growth and development in recent years. Recent advances in this industry have focused on reducing its economic and environmental costs, for example, the substitution of protein sources in fish feed. Plant-based proteins are now a common approach, and while it is a greener alternative to animal-based proteins, there are some disadvantages, such as their putative content and intoxicants such as mycotoxins. These are naturally occurring plant contaminants, and their exposure in fish can cause health problems, stunted growth or even death, resulting in economic losses for the producers and health concerns for the consumers. Different works have demonstrated the presence of both AFB1 (aflatoxin B1) and ENNB1 (enniatin B1) in fish feed and their capacity to be absorbed and bioaccumulate in the fish organism after digestion, further reaching humans through fish ingestion. The aim of this work was to evaluate the bioaccessibility of both mycotoxins in samples of Sparus aurata muscle using a static digestion model based on the INFOGEST protocol. The samples were subjected to different cooking procedures – raw, grilled and fried – and different seasonings – none, thyme and ginger – in order to evaluate their potential reduction effect on mycotoxins bioaccessibility, followed by the evaluation of the intestinal transport of both compounds with an in vitro cell model composed of Caco-2/HT-29 co-culture monolayers, simulating the human intestinal epithelium. The bioaccessible fractions obtained in the digestion studies were used in the transport studies for a more realistic approach to bioavailability evaluation. Results demonstrated the effect of the use of different cooking procedures and seasoning on the toxin's bioavailability. Sparus aurata was chosen in this study for its large production in aquaculture and high consumption in Europe. Also, with the continued evolution of fish farming practices and more common usage of novel feed ingredients based on plants, there is a growing concern about less studied contaminants in aquaculture and their consequences for human health. In pair with greener advances in this industry, there is a convergence towards alternative research methods, such as in vitro applications. In the case of bioavailability studies, both in vitro digestion protocols and intestinal transport assessment are excellent alternatives to in vivo studies. These methods provide fast, reliable and comparable results without ethical restraints.

Keywords: AFB1, aquaculture, bioaccessibility, ENNB1, intestinal transport.

Procedia PDF Downloads 37
150 Influence of Dietary Boron on Gut Absorption of Nutrients, Blood Metabolites and Tissue Pathology

Authors: T. Vijay Bhasker, N. K. S Gowda, P. Krishnamoorthy, D. T. Pal, A. K. Pattanaik, A. K. Verma

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Boron (B) is a newer trace element and its biological importance and dietary essentiality is unclear in animals. The available literature suggests its putative role in bone mineralization, antioxidant status and steroid hormone synthesis. A feeding trial was conducted in Wister strain (Rattus norvegicus) albino rats for duration of 90 days. A total of 84 healthy weaned (3-4 weeks) experimental rats were randomly divided into 7 dietary groups (4 replicates of three each) viz., A (Basal diet/ Control), B (Basal diet + 5 ppm B), C (Basal diet + 10 ppm B), D (Basal diet + 20 ppm B), E (Basal diet + 40 ppm B), F (Basal diet-Ca 50%), G (Basal diet-Ca 50% + 40 ppm B). Dietary level of calcium (Ca) was maintained at two levels, 100% and 50% of requirement. Sodium borate was used as source of boron along with other ingredients of basal diet while preparing the pelletized diets. All the rats were kept in proper ventilated laboratory animal house maintained at temperature (23±2º C) and humidity (50 to 70%). At the end of experiment digestibility trial was conducted for 5 days to estimate nutrient digestibility and gut absorption of minerals. Eight rats from each group were sacrificed to collect the vital organs (liver, kidney and spleen) to study histopathology. Blood sample was drawn by heart puncture to determine biochemical profile. The average daily feed intake (g/rat/day), water intake (ml/rat/day) and body weight gain (g/rat/day) were similar among the dietary groups. The digestibility (%) of organic matter and crude fat were significantly improved (P < 0.05) was by B supplementation. The gut absorption (%) Ca was significantly increased (P < 0.01) in B supplemented groups compared to control. However, digestibility of dry matter and crude protein, gut absorption of magnesium and phosphorus showed a non-significant increasing trend with B supplementation. The gut absorption (%) of B (P < 0.01) was significantly lowered (P<0.05) in supplemented groups compared to un-supplemented ones. The serum level of triglycerides (mg/dL), HDL-cholesterol (mg/dL) and alanine transaminase (IU/L) were significantly lowered (P < 0.05) in B supplemented groups. While serum level of glucose (mg/dL) and alkaline phosphatase (KA units) showed a non-significant decreasing trend with B supplementation. However the serum levels of total cholesterol (mg/dL) and aspartate transaminase (IU/L) were similar among dietary groups. The histology sections of kidney and spleen revealed no significant changes among the dietary groups and were observed to be normal in anatomical architecture. However, the liver histology revealed cell degenerative changes with vacuolar degeneration and nuclear condensation in Ca deficient groups. But the comparative degenerative changes were mild in 40 ppm B supplemented Ca deficient group. In conclusion, dietary supplementation of graded levels of boron in rats had a positive effect on metabolism and health by improving nutrient digestibility and gut absorption of Ca. This indicates the beneficial role of dietary boron supplementation.

Keywords: boron, calcium, nutrient utilization, histopathology

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149 Lesbians, Gays and Bisexuals of Botswana: Progressive Steps by the Botswana Court of Appeal towards Recognition and Advancement of Fundamental Human Rights of the Most Vulnerable within Society

Authors: Tashwill Esterhuizen

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Throughout Africa, several countries continue to have laws which criminalise same-sex sexual activities, which increases the vulnerability of the LGBT community to stigma, discrimination, and persecution. These criminal provisions often form the basis upon which states deny LGBT activists the right to freely associate with other like-minded individuals and form organizations that protect their interests and advocate for the rights and aspirations of the LGBT community. Over the past year, however, there has been significant progress in the advancement of universal, fundamental rights of LGBT persons throughout Africa. In many instances, these advancements came about through the bravery of activists who have publically insisted (in environments where same-sex sexual practices are criminalised) that their rights should be respected. Where meaningful engagement with the State was fruitless, activists took their plight to the judiciary and have successfully sought to uphold the fundamental rights of LGBT persons, paving the way for a more inclusive and tolerant society. Litigation Progress: Botswana is a prime example. For several years, the State denied a group of LGBT activists their right to freely associate and form their organisation Lesbians, Gays, and Bisexuals of Botswana (LEGABIBO), which aimed to promote the interests of the LGBT community in Botswana. In March 2016, the Botswana Court of Appeal found that the government’s refusal to register LEGABIBO violated the activists’ right to associate freely. The Court held that the right freedom of association applies to all persons regardless of their sexual orientation or gender identity. It does not matter that the views of the organisation are unpopular or unacceptable amongst the majority. In particular, the Court rejected the government of Botswana’s contention that registering LEGABIBO would disturb public peace and is contrary to public morality. Quite remarkably, the Court of Appeal recognised that while LGBT individuals are a minority group within the country, they are nonetheless persons entitled to constitutional protections of their dignity, regardless of whether they are unacceptable to others on religious or any other grounds. Furthermore, the Court held that human rights and fundamental freedoms are granted to all, including criminals or social outcasts because the denial of an individual’s humanity is the denial of their human dignity. This is crucial observation by the Court of Appeal, as once it is accepted that human rights apply to all human beings, then it becomes much easier for vulnerable groups to assert their own rights. Conclusion: The Botswana Court of Appeal decision, therefore, represents significant progress in the promotion of the rights of lesbian, gay, bisexual and transgender persons. The judgment has broader implications for many other countries which do not provide recognition of sexual minorities. It highlights the State’s duty to uphold basic rights and to ensure dignity, tolerance, and acceptance for marginalised persons.

Keywords: acceptance, freedom of association, freedom of expression, fundamental rights and freedoms, gender identity, human rights are universal, inclusive, inherent human dignity, progress, sexual orientation, tolerance

Procedia PDF Downloads 216
148 Writing the Roaming Female Self: Identity and Romantic Selfhood in Mary Wollstonecraft’s Letters Written during a Short Stay in Sweden, Denmark, and Norway (1796)

Authors: Kalyani Gandhi

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The eighteenth century in Britain saw a great burst of activity in writing (letters, journals, newspapers, essays); often these modes of writing had a public-spirited bent in-step with the prevailing intellectual atmosphere. Mary Wollstonecraft was one of the leading intellectuals of that period who utilized letter writing to convey her thoughts on the exciting political developments of the late eighteenth century. Fusing together her anxieties and concerns about humanity in general and herself in particular, Wollstonecraft’s views of the world around her are filtered through the lens of her subjectivity. Thus, Wollstonecraft’s letters covered a wide range of topics on both the personal and political level (for the two are often entwined in Wollstonecraft’s characteristic style of analysis) such as sentiment, gender, nature, peasantry, the class system, the legal system, political duties and rights of both rulers and subjects, death, immortality, religion, family and education. Therefore, this paper intends to examine the manner in which Wollstonecraft utilizes letter-writing to constitute and develop Romantic self-hood, understand the world around her and illustrate her ideas on the political and social happenings in Europe. The primary text analyzed will be Mary Wollstonecraft's Letters Written During a Short Stay in Sweden, Denmark and Norway (1796) and the analysis of this text will be supplemented by researching 18th-century British letter writing culture, with a special emphasis on the epistolary habits of women. Within this larger framework, this paper intends to examine the manner in which this hybrid of travel and epistolary writing aided Mary Wollstonecraft's expression on Romantic selfhood and how it was complicated by ideas of gender. This paper reveals Wollstonecraft's text to be wrought with anxiety about the world around her and within her; thus, the personal-public nature of the epistolary format particularly suits her characteristic point of view that looks within and without. That is to say, Wollstonecraft’s anxieties about gender and self, are as much about the women she sees in the world around her as much as they are about her young daughter and herself. Wollstonecraft constantly explores and examines this anxiety within the different but interconnected realms of politics, economics, history and society. In fact, it is her complex technique of entwining these aforementioned concerns with a closer look at interpersonal relationships among men and women (she often mentions specific anecdotes and instances) that make Wollstonecraft's Letters so engaging and insightful. Thus, Wollstonecraft’s Letters is an exemplar of British Romantic writing due to the manner in which it explores the bond between the individual and society. Mary Wollstonecraft's nuances this exploration by incorporating her concerns about women and the playing out of gender in society. Thus, Wollstonecraft’s Letters is an invaluable contribution to the field of British Romanticism, particularly as it offers crucial insight on female Romantic writing that can broaden and enrich the current academic understanding of the field.

Keywords: British romanticism, letters, feminism, travel writing

Procedia PDF Downloads 186
147 Tip60 Histone Acetyltransferase Activators as Neuroepigenetic Therapeutic Modulators for Alzheimer’s Disease

Authors: Akanksha Bhatnagar, Sandhya Kortegare, Felice Elefant

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Context: Alzheimer's disease (AD) is a neurodegenerative disorder that is characterized by progressive cognitive decline and memory loss. The cause of AD is not fully understood, but it is thought to be caused by a combination of genetic, environmental, and lifestyle factors. One of the hallmarks of AD is the loss of neurons in the hippocampus, a brain region that is important for memory and learning. This loss of neurons is thought to be caused by a decrease in histone acetylation, which is a process that regulates gene expression. Research Aim: The research aim of the study was to develop mall molecule compounds that can enhance the activity of Tip60, a histone acetyltransferase that is important for memory and learning. Methodology/Analysis: The researchers used in silico structural modeling and a pharmacophore-based virtual screening approach to design and synthesize small molecule compounds strongly predicted to target and enhance Tip60’s HAT activity. The compounds were then tested in vitro and in vivo to assess their ability to enhance Tip60 activity and rescue cognitive deficits in AD models. Findings: The researchers found that several of the compounds were able to enhance Tip60 activity and rescue cognitive deficits in AD models. The compounds were also developed to cross the blood-brain barrier, which is an important factor for the development of potential AD therapeutics. Theoretical Importance: The findings of this study suggest that Tip60 HAT activators have the potential to be developed as therapeutic agents for AD. The compounds are specific to Tip60, which suggests that they may have fewer side effects than other HDAC inhibitors. Additionally, the compounds are able to cross the blood-brain barrier, which is a major hurdle for the development of AD therapeutics. Data Collection: The study collected data from a variety of sources, including in vitro assays and animal models. The in vitro assays assessed the ability of compounds to enhance Tip60 activity using histone acetyltransferase (HAT) enzyme assays and chromatin immunoprecipitation assays. Animal models were used to assess the ability of the compounds to rescue cognitive deficits in AD models using a variety of behavioral tests, including locomotor ability, sensory learning, and recognition tasks. The human clinical trials will be used to assess the safety and efficacy of the compounds in humans. Questions: The question addressed by this study was whether Tip60 HAT activators could be developed as therapeutic agents for AD. Conclusions: The findings of this study suggest that Tip60 HAT activators have the potential to be developed as therapeutic agents for AD. The compounds are specific to Tip60, which suggests that they may have fewer side effects than other HDAC inhibitors. Additionally, the compounds are able to cross the blood-brain barrier, which is a major hurdle for the development of AD therapeutics. Further research is needed to confirm the safety and efficacy of these compounds in humans.

Keywords: Alzheimer's disease, cognition, neuroepigenetics, drug discovery

Procedia PDF Downloads 49
146 Accelerating Personalization Using Digital Tools to Drive Circular Fashion

Authors: Shamini Dhana, G. Subrahmanya VRK Rao

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The fashion industry is advancing towards a mindset of zero waste, personalization, creativity, and circularity. The trend of upcycling clothing and materials into personalized fashion is being demanded by the next generation. There is a need for a digital tool to accelerate the process towards mass customization. Dhana’s D/Sphere fashion technology platform uses digital tools to accelerate upcycling. In essence, advanced fashion garments can be designed and developed via reuse, repurposing, recreating activities, and using existing fabric and circulating materials. The D/Sphere platform has the following objectives: to provide (1) An opportunity to develop modern fashion using existing, finished materials and clothing without chemicals or water consumption; (2) The potential for an everyday customer and designer to use the medium of fashion for creative expression; (3) A solution to address the global textile waste generated by pre- and post-consumer fashion; (4) A solution to reduce carbon emissions, water, and energy consumption with the participation of all stakeholders; (5) An opportunity for brands, manufacturers, retailers to work towards zero-waste designs and as an alternative revenue stream. Other benefits of this alternative approach include sustainability metrics, trend prediction, facilitation of disassembly and remanufacture deep learning, and hyperheuristics for high accuracy. A design tool for mass personalization and customization utilizing existing circulating materials and deadstock, targeted to fashion stakeholders will lower environmental costs, increase revenues through up to date upcycled apparel, produce less textile waste during the cut-sew-stitch process, and provide a real design solution for the end customer to be part of circular fashion. The broader impact of this technology will result in a different mindset to circular fashion, increase the value of the product through multiple life cycles, find alternatives towards zero waste, and reduce the textile waste that ends up in landfills. This technology platform will be of interest to brands and companies that have the responsibility to reduce their environmental impact and contribution to climate change as it pertains to the fashion and apparel industry. Today, over 70% of the $3 trillion fashion and apparel industry ends up in landfills. To this extent, the industry needs such alternative techniques to both address global textile waste as well as provide an opportunity to include all stakeholders and drive circular fashion with new personalized products. This type of modern systems thinking is currently being explored around the world by the private sector, organizations, research institutions, and governments. This technological innovation using digital tools has the potential to revolutionize the way we look at communication, capabilities, and collaborative opportunities amongst stakeholders in the development of new personalized and customized products, as well as its positive impacts on society, our environment, and global climate change.

Keywords: circular fashion, deep learning, digital technology platform, personalization

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145 Influence of Iron Content in Carbon Nanotubes on the Intensity of Hyperthermia in the Cancer Treatment

Authors: S. Wiak, L. Szymanski, Z. Kolacinski, G. Raniszewski, L. Pietrzak, Z. Staniszewska

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The term ‘cancer’ is given to a collection of related diseases that may affect any part of the human body. It is a pathological behaviour of cells with the potential to undergo abnormal breakdown in the processes that control cell proliferation, differentiation, and death of particular cells. Although cancer is commonly considered as modern disease, there are beliefs that drastically growing number of new cases can be linked to the extensively prolonged life expectancy and enhanced techniques for cancer diagnosis. Magnetic hyperthermia therapy is a novel approach to cancer treatment, which may greatly contribute to higher efficiency of the therapy. Employing carbon nanotubes as nanocarriers for magnetic particles, it is possible to decrease toxicity and invasiveness of the treatment by surface functionalisation. Despite appearing in recent years, magnetic particle hyperthermia has already become of the highest interest in the scientific and medical environment. The reason why hyperthermia therapy brings so much hope for future treatment of cancer lays in the effect that it produces in malignant cells. Subjecting them to thermal shock results in activation of numerous degradation processes inside and outside the cell. The heating process initiates mechanisms of DNA destruction, protein denaturation and induction of cell apoptosis, which may lead to tumour shrinkage, and in some cases, it may even cause complete disappearance of cancer. The factors which have the major impact on the final efficiency of the treatment include temperatures generated inside the tissues, time of exposure to the heating process, and the character of an individual cancer cell type. The vast majority of cancer cells is characterised by lower pH, persistent hypoxia and lack of nutrients, which can be associated to abnormal microvasculature. Since in healthy tissues we cannot observe presence of these conditions, they should not be seriously affected by elevation of the temperature. The aim of this work is to investigate the influence of iron content in iron filled Carbon Nanotubes on the desired nanoparticles for cancer therapy. In the article, the development and demonstration of the method and the model device for hyperthermic selective destruction of cancer cells are presented. This method was based on the synthesis and functionalization of carbon nanotubes serving as ferromagnetic material nanocontainers. The methodology of the production carbon- ferromagnetic nanocontainers (FNCs) includes the synthesis of carbon nanotubes, chemical, and physical characterization, increasing the content of a ferromagnetic material and biochemical functionalization involving the attachment of the key addresses. The ferromagnetic nanocontainers were synthesised in CVD and microwave plasma system. The research work has been financed from the budget of science as a research project No. PBS2/A5/31/2013.

Keywords: hyperthermia, carbon nanotubes, cancer colon cells, radio frequency field

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144 Nutritional Genomics Profile Based Personalized Sport Nutrition

Authors: Eszter Repasi, Akos Koller

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Our genetic information determines our look, physiology, sports performance and all our features. Maximizing the performances of athletes have adopted a science-based approach to the nutritional support. Nowadays genetics studies have blended with nutritional sciences, and a dynamically evolving, new research field have appeared. Nutritional genomics is needed to be used by nutritional experts. This is a recent field of nutritional science, which can provide a solution to reach the best sport performance using correlations between the athlete’s genome, nutritions, molecules, included human microbiome (links between food, microbiome and epigenetics), nutrigenomics and nutrigenetics. Nutritional genomics has a tremendous potential to change the future of dietary guidelines and personal recommendations. Experts need to use new technology to get information about the athletes, like nutritional genomics profile (included the determination of the oral and gut microbiome and DNA coded reaction for food components), which can modify the preparation term and sports performance. The influence of nutrients on the genes expression is called Nutrigenomics. The heterogeneous response of gene variants to nutrients, dietary components is called Nutrigenetics. The human microbiome plays a critical role in the state of health and well-being, and there are more links between food or nutrition and the human microbiome composition, which can develop diseases and epigenetic changes as well. A nutritional genomics-based profile of athletes can be the best technic for a dietitian to make a unique sports nutrition diet plan. Using functional food and the right food components can be effected on health state, thus sports performance. Scientists need to determine the best response, due to the effect of nutrients on health, through altering genome promote metabolites and result changes in physiology. Nutritional biochemistry explains why polymorphisms in genes for the absorption, circulation, or metabolism of essential nutrients (such as n-3 polyunsaturated fatty acids or epigallocatechin-3-gallate), would affect the efficacy of that nutrient. Controlled nutritional deficiencies and failures, prevented the change of health state or a newly discovered food intolerance are observed by a proper medical team, can support better sports performance. It is important that the dietetics profession informed on gene-diet interactions, that may be leading to optimal health, reduced risk of injury or disease. A special medical application for documentation and monitoring of data of health state and risk factors can uphold and warn the medical team for an early action and help to be able to do a proper health service in time. This model can set up a personalized nutrition advice from the status control, through the recovery, to the monitoring. But more studies are needed to understand the mechanisms and to be able to change the composition of the microbiome, environmental and genetic risk factors in cases of athletes.

Keywords: gene-diet interaction, multidisciplinary team, microbiome, diet plan

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143 Melancholia, Nostalgia: Bernardo Soares after Fernando Pessoa

Authors: Maria de Fátima Lambert

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Bernardo Soares is one of Fernando Pessoa' several heteronyms (and "half-heteronyms"). Perhaps the one that brought together the majority of his qualities and characters of self-identity within the famous inner-persona-alter-diversity. The Book of Disquiet by Bernardo Soares was released in 1983, consisting of ontological remarks caught by an obsessive inquiring about self-existence. The book became a highly valuable substance when focusing upon the philosophical grounds of Pessoa's aesthetics. For sure, we cannot consider a single aesthetic, admitting that each heteronym has its own particular one, developed after different principles and convictions. Regarding Bernardo Soares, his thought arises from sequenced self-clues expressing peculiar existential doubtless presented as certainties -and vice-versa. His written self-search-images are reported, molding the painful awareness of existence through the discredited tolerance of any conclusive dialogue with others. Given the nature of Soares’ [maybe] unfinished writings, it is obvious that he headed far from his self-insurance-capsule: the office, bedroom, or even the walkscapes through Lisbon. The idea of travel/journey is one of the most relevant when recognizing his profound - although undercover - anguish as melancholy and nostalgia. In Bernardo Soares, Aesthetics is taken agonizingly, grounded upon discreet poetic phraseology and terms. His poetical awareness developed compulsive titles such "Aesthetics of Indifference", "Aesthetics of Discouragement". Soares' Aesthetics emerges directly from oneself, understanding art as inner acts and living experienced issues. Art is not freed from the intellectual expression of oneself emotions. The Disquiet Book is an existential nightmare nourished by everyday life, single written thoughts, balanced by melancholia, nostalgia, and distress. One might wonder if it was dreams that guided his fictional literary persona or the narrow facts of life itself. Along with his endless disquiet writing, Pessoa’s semi-heteronymous traveled without physically going anywhere. The complexity of inner existence is fulfilled by lonely mental walks and travels, as in two texts titled The Never Accomplished Journey. Although we also can consider other fragments, these are the deepest reflections about travelling. Let’s recall that Fernando Pessoa’s ortonyms writings -poems and essays- also addressed this issue from a philosophical perspective. We believe that this theme is one of the meaningful concepts for featuring the main principles of his aesthetics. As we know, Fernando Pessoa did not travel to foreign countries (or in Portugal), except for the journey, with his family, from Lisbon to South Africa (as a child) and, some years later, the return back to Lisbon. One may wonder why the poet never undertook other journeys. Maybe due to a disbelief in moving away from his comfort zone or due to the fear of becoming addicted to endless travels and the loss of his convenient self-closeness. In The Book of Disquiet, the poet shared his internal visions of the outer world but mainly visualizing his deepest enigmas and experiences -so strongly incorporated into reality and fiction.

Keywords: aesthetic principles, Bernardo Soares, Fernando Pessoa , melancholia, nostalgia, non-accomplished travel, The Book of Disquiet

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142 Fodder Production and Livestock Rearing in Relation to Climate Change and Possible Adaptation Measures in Manaslu Conservation Area, Nepal

Authors: Bhojan Dhakal, Naba Raj Devkota, Chet Raj Upreti, Maheshwar Sapkota

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A study was conducted to find out the production potential, nutrient composition, and the variability of the most commonly available fodder trees along with the varying altitude to help optimize the dry matter requirement during winter lean period. The study was carried out from March to June, 2012 in Lho and Prok Village Development Committee of Manaslu Conservation Area (MCA), located in Gorkha district of Nepal. The other objective of the research was to learn the impact of climate change on livestock production linking it with feed availability. The study was conducted in two parts: social and biological. Accordingly, a households (HHs) survey was conducted to collect primary data from 70 HHs, focusing on the perception of respondents on impacts of climatic variability on the feeding management. The next part consisted of understanding yield potential and nutrient composition of the four most commonly available fodder trees (M. azedirach, M. alba, F. roxburghii, F. nemoralis), within two altitudes range: (1500-2000 masl and 2000-2500 masl) by using a RCB design in 2*4 factorial combination of treatments, each replicated four times. Results revealed that majority of the farmers perceived the change in climatic phenomenon more severely within the past five years. Farmers were using different adaptation technologies such as collection of forage from jungle, reducing unproductive animals, fodder trees utilization, and crop by product feeding at feed scarcity period. Ranking of the different fodder trees on the basis of indigenous knowledge and experiences revealed that F. roxburghii was the best-preferred fodder tree species (index value 0.72) in terms overall preferability whereas M. azedirach had highest growth and productivity (index value 0.77), F. roxburghii had highest adoptability (index value 0.69) and palatability (index value 0.69) as well. Similarly, fresh yield and dry matter yield of the each fodder trees was significant (P < 0.01) between the altitude and within species. Fodder trees yield analysis revealed that the highest dry matter (DM) yield (28 kg/tree) was obtained for F. roxburghii but that remained statistically similar (P > 0.05) to the other treatment. On the other hand, most of the parameters: ether extract (EE), acid detergent lignin (ADL), acid detergent fibre (ADF), cell wall digestibility (CWD), relative digestibility (RD), digestible nutrient (TDN), and Calcium (Ca) among the treatments were highly significant (P < 0.01). This indicates the scope of introducing productive and nutritive fodder trees species even at the high altitude to help reduce fodder scarcity problem during winter. The finding also revealed the scope of promoting all available local fodder trees species as crude protein content of these species were similar.

Keywords: fodder trees, yield potential, climate change, nutrient composition

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141 Mesenchymal Stem Cells on Fibrin Assemblies with Growth Factors

Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

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Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

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