Search results for: protein masked representation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3618

Search results for: protein masked representation

2898 The Effects of Seasonal Variation on the Microbial-N Flow to the Small Intestine and Prediction of Feed Intake in Grazing Karayaka Sheep

Authors: Mustafa Salman, Nurcan Cetinkaya, Zehra Selcuk, Bugra Genc

Abstract:

The objectives of the present study were to estimate the microbial-N flow to the small intestine and to predict the digestible organic matter intake (DOMI) in grazing Karayaka sheep based on urinary excretion of purine derivatives (xanthine, hypoxanthine, uric acid, and allantoin) by the use of spot urine sampling under field conditions. In the trial, 10 Karayaka sheep from 2 to 3 years of age were used. The animals were grazed in a pasture for ten months and fed with concentrate and vetch plus oat hay for the other two months (January and February) indoors. Highly significant linear and cubic relationships (P<0.001) were found among months for purine derivatives index, purine derivatives excretion, purine derivatives absorption, microbial-N and DOMI. Through urine sampling and the determination of levels of excreted urinary PD and Purine Derivatives / Creatinine ratio (PDC index), microbial-N values were estimated and they indicated that the protein nutrition of the sheep was insufficient. In conclusion, the prediction of protein nutrition of sheep under the field conditions may be possible with the use of spot urine sampling, urinary excreted PD and PDC index. The mean purine derivative levels in spot urine samples from sheep were highest in June, July and October. Protein nutrition of pastured sheep may be affected by weather changes, including rainfall. Spot urine sampling may useful in modeling the feed consumption of pasturing sheep. However, further studies are required under different field conditions with different breeds of sheep to develop spot urine sampling as a model.

Keywords: Karayaka sheep, spot sampling, urinary purine derivatives, PDC index, microbial-N, feed intake

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2897 Identification of Nutrient Sensitive Signaling Pathways via Analysis of O-GlcNAcylation

Authors: Michael P. Mannino, Gerald W. Hart

Abstract:

The majority of glucose metabolism proceeds through glycolytic pathways such as glycolysis or pentose phosphate pathway, however, about 5% is shunted through the hexosamine biosynthetic pathway, producing uridine diphosphate N-acetyl glucosamine (UDP-GlcNAc). This precursor can then be incorporated into complex oligosaccharides decorating the cell surface or remain as an intracellular post-translational-modification (PTM) of serine/threonine residues (O-GlcNAcylation, OGN), which has been identified on over 4,000 cytosolic or nuclear proteins. Intracellular OGN has major implications on cellularprocesses, typically by modulating protein localization, protein-protein interactions, protein degradation, and gene expression. Additionally, OGN is known to have an extensive cross-talk with phosphorylation, be in a competitive or cooperative manner. Unlike other PTMs there are only two cycling enzymes that are capable of adding or removing the GlcNAc moiety, O-linked N-aceytl glucosamine Transferase (OGT) and O-linked N-acetyl glucoamidase (OGA), respectively. The activity of OGT has been shown to be sensitive to cellular UDP-GlcNAc levels, even changing substrate affinity. Owing to this and that the concentration of UDP-GlcNAc is related to the metabolisms of glucose, amino acid, fatty acid, and nucleotides, O-GlcNAc is often referred to as a nutrient sensing rheostat. Indeed OGN is known to regulate several signaling pathways as a result of nutrient levels, such as insulin signaling. Dysregulation of OGN is associated with several disease states such as cancer, diabetes, and neurodegeneration. Improvements in glycomics over the past 10-15 years has significantly increased the OGT substrate pool, suggesting O-GlcNAc’s involvement in a wide variety of signaling pathways. However, O-GlcNAc’s role at the receptor level has only been identified in a case-by-case basis of known pathways. Examining the OGN of the plasma membrane (PM) may better focus our understanding of O-GlcNAc-effected signaling pathways. In this current study, PM fractions were isolated from several cell types via ultracentrifugation, followed by purification and MS/MS analysis in several cell lines. This process was repeated with or without OGT/OGA inhibitors or with increased/decreased glucose levels in media to ascertain the importance of OGN. Various pathways are followed up on in more detailed studies employing methods to localize OGN at the PM specifically.

Keywords: GlcNAc, nutrient sensitive, post-translational-modification, receptor

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2896 Of Rites of Narration and Representation of Orient and Occident in Thomas Heywood's Fair Maid of the West

Authors: Tarik Bouguerba

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Thomas Heywood was an outstanding, prolific playwright of the period, writing both in prose and verse. Unlike Shakespeare in particular, Heywood could be considered as a playwright who was well informed about Morocco and wrote in greater detail about a possible dialogue among cultures. As it is a historical platform for power relations, The Fair Maid of the West recalled the heroism and excitement of English counterattacks against Spain in the Post-Armada period. This paper therefore pins down the acts of narration and representation of Morocco and Moroccans and examines how the Occident has contributed to the production of the Orient and finally attests to the metamorphosis the plot undergoes in Part I and Part II. As an adventure play, The Fair Maid of the West teaches about, informs of and confirms the existing patterns of virtue in European voyagers and at the same time it asserts how honor and chastity are European par excellence whereas villainy and wickedness are Oriental assets. Once taken captive, these virtues and traits are put into task as the plot disentangles. This paper also examines how the play in both parts generates a whole history of stereotypes about Morocco and unexpectedly subverts this stereotype; such a biased mode of narration of the Orient the playwright took up at first was played down at a later phase in the narrative.

Keywords: Heywood, Occident, Orientalism, Stereotype, Virtue

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2895 The Effect of Fermentation and Germination on the Nutrient and Antinutrient Composition of Lima Bean (Phaseolus lunatus) Flour

Authors: P. N. Okeke

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Fermentation and germination of legumes have been an ancient practice. In this study, the influence of fermentation and germination on the chemical properties of Lima bean (Phaseolus lunatus) flour were evaluated. The flours were analyzed for their proximate and mineral composition, using the standard assay methods. The result showed that fermentation and germination increased the moisture, protein and ash content of the flours while fiber, fat and carbohydrate were decreased. The protein level of fermented and germinated lima bean increased from 21.06–26.60%. The minerals: iron, copper, zinc, and phosphorous increased due to germination and fermentation. The phytate and tannin levels were drastically reduced in both the fermented and germinated flours. The result of this study revealed that fermentation and germination makes the nutrient in lima beans more accessible as it reduces the anti-nutrients. It is therefore recommended that lima bean be process accordingly for richer and more bio-availability of the nutrients.

Keywords: nutrient, anti-nutrient, fermented, germinated, lima bean flour

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2894 Identification of Significant Genes in Rheumatoid Arthritis, Melanoma Metastasis, Ulcerative Colitis and Crohn’s Disease

Authors: Krishna Pal Singh, Shailendra Kumar Gupta, Olaf Wolkenhauer

Abstract:

Background: Our study aimed to identify common genes and potential targets across the four diseases, which include rheumatoid arthritis, melanoma metastasis, ulcerative colitis, and Crohn’s disease. We used a network and systems biology approach to identify the hub gene, which can act as a potential target for all four disease conditions. The regulatory network was extracted from the PPI using the MCODE module present in Cytoscape. Our objective was to investigate the significance of hub genes in these diseases using gene ontology and KEGG pathway enrichment analysis. Methods: Our methodology involved collecting disease gene-related information from DisGeNET databases and performing protein-protein interaction (PPI) network and core genes screening. We then conducted gene ontology and KEGG pathway enrichment analysis. Results: We found that IL6 plays a critical role in all disease conditions and in different pathways that can be associated with the development of all four diseases. Conclusions: The theoretical importance of our research is that we employed various systems and structural biology techniques to identify a crucial protein that could serve as a promising target for treating multiple diseases. Our data collection and analysis procedures involved rigorous scrutiny, ensuring high-quality results. Our conclusion is that IL6 plays a significant role in all four diseases, and it can act as a potential target for treating them. Our findings may have important implications for the development of novel therapeutic interventions for these diseases.

Keywords: melanoma metastasis, rheumatoid arthritis, inflammatory bowel diseases, integrated bioinformatics analysis

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2893 Issues in the Learning and Construction of a National Music Identity in Multiracial Malaysia: Diversity, Complexity, and Contingency

Authors: Loo Fung Ying, Loo Fung Chiat

Abstract:

The formation of a musical identity that shapes the nation in this multiracial country reveals many complexities, conundrums, and contingencies. Creativity and identity formation at the level of an individual or a collective group further diversified musical expression, representation, and style, which has led to an absence of regularities. In addition, ‘contemporizing accretion,’ borrowing a term used by Schnelle in theology (2009), further complicates musical identity, authenticity, conception, and realization. Thus, in this paper, we attempt to define the issues surrounding the teaching and learning of the multiracial Malaysian national music identity. We also discuss unnecessary power hierarchies, interracial conflicts, and sentiments in the construct of a multiracial national music identity by referring to genetic origins, the evolution of music, and the neglected issues of representation and reception at a global level from a diachronic perspective. Lastly, by synthesizing Ladson-Billings, Gay, Kruger, and West-Burns’s culturally relevant/responsive pedagogical theories, we discuss possible analytic tools for consideration that are more multiculturally relevant and responsive for the teaching, learning, and construction of a multiracial Malaysian national music identity.

Keywords: Malaysia, music, multiracial, national music identity, culturally relevant/responsive pedagogy

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2892 When the Rubber Hits the Road: The Enactment of Well-Intentioned Language Policy in Digital vs. In Situ Spaces on Washington, DC Public Transportation

Authors: Austin Vander Wel, Katherin Vargas Henao

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Washington, DC, is a city in which Spanish, along with several other minority languages, is prevalent not only among tourists but also those living within city limits. In response to this linguistic diversity and DC’s adoption of the Language Access Act in 2004, the Washington Metropolitan Area Transit Authority (WMATA) committed to addressing the need for equal linguistic representation and established a five-step plan to provide the best multilingual information possible for public transportation users. The current study, however, strongly suggests that this de jure policy does not align with the reality of Spanish’s representation on DC public transportation–although perhaps doing so in an unexpected way. In order to investigate Spanish’s de facto representation and how it contrasts with de jure policy, this study implements a linguistic landscapes methodology that takes critical language-policy as its theoretical framework (Tollefson, 2005). Specifically concerning de facto representation, it focuses on the discrepancies between digital spaces and the actual physical spaces through which users travel. These digital vs. in situ conditions are further analyzed by separately addressing aural and visual modalities. In digital spaces, data was collected from WMATA’s website (visual) and their bilingual hotline (aural). For in situ spaces, both bus and metro areas of DC public transportation were explored, with signs comprising the visual modality and recordings, driver announcements, and interactions with metro kiosk workers comprising the aural modality. While digital spaces were considered to successfully fulfill WMATA’s commitment to representing Spanish as outlined in the de jure policy, physical spaces show a large discrepancy between what is said and what is done, particularly regarding the bus system, in addition to the aural modality overall. These discrepancies in situ spaces place Spanish speakers at a clear disadvantage, demanding additional resources and knowledge on the part of residents with limited or no English proficiency in order to have equal access to this public good. Based on our critical language-policy analysis, while Spanish is represented as a right in the de jure policy, its implementation in situ clearly portrays Spanish as a problem since those seeking bilingual information can not expect it to be present when and where they need it most (Ruíz, 1984; Tollefson, 2005). This study concludes with practical, data-based steps to improve the current situation facing DC’s public transportation context and serves as a model for responding to inadequate enactment of de jure policy in other language policy settings.

Keywords: Urban landscape, language access, critical-language policy, spanish, public transportation

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2891 Chemotactic Behaviour of Human Mesenchymal Stem Cells in Response to Silicate Substituted Hydroxyapatite

Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson

Abstract:

Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.

Keywords: cell migration, hMSCs, SiHA, transwell migration system

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2890 Application of Microparticulated Whey Proteins in Reduced-Fat Yogurt through Hot-Extrusion: Influence on Physicochemical and Sensory Properties

Authors: M. K. Hossain, J. Keidel, O. Hensel, M. Diakite

Abstract:

Fat reduced dairy products are holding a potential market due to health reason. Due to less creamy, and pleasantness, reduced and/or low-fat dairy products are getting less consumer acceptance whereas the fat molecule provides smooth, creamy and a pleasant mouthfeel in dairy products especially yogurt & ice cream. This study was aimed to investigate whether the application of microparticulated whey proteins (MWPs) processed by extrusion cooking, the reduced fat yogurt can achieve similar or higher creaminess compared to whole milk (3.8% fat) and skimmed milk (0.5% fat) yogurt. Full cream and skimmed milk were used to prepare natural stirred yogurt, as well as the dry matter content, also adjusted up to 16% with skimmed milk powder. Whey protein concentrates (WPC80) were used to produce MWPs in particle size of d50 > 5 µm, d50 3<5 µm and d50 < 3 µm through the hot-extrusion process with a screw speed of 400, 600 and 1000 rpm respectively. Furthermore, the commercially available microparticulated whey protein called Simplesse® was also applied in order to compare with extruded MWPs. The rheological and sensory properties of yogurt were assessed, and data were analyzed statistically. The applications of extruded MWPs with 600 and 1000 rpm were achieved significantly (p < 0.05) higher creaminess and preference compared to the whole and skimmed milk yogurt whereas, 400 rpm got lower preference. On the other hand, Simplesse® obtained the lowest creaminess and preference compared to other yogurts, although the contribution of dry matter in yogurt was same as extruded MWPs. The creaminess and viscosities were strongly (r = 0.62) correlated, furthermore, the viscosity from sensory evaluation and the dynamic viscosity of yogurt was also significantly (r = 0.72) correlated which clarifies that the performance of sensory panelists as well as the quality of the products.

Keywords: microparticulation, hot-extrusion, reduced-fat yogurt, whey protein concentrate

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2889 Genesis and Achievements of Madhesh Movement in Nepal

Authors: Deepak Chaudhary

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The main objective of the study is to explore the genesis and achievements of the Madhesh movement. Madhesh Movement is a social movement that brought massive political changes and contributed a lot to the nation-building process in the modern history of Nepal. This movement erupted in January 2007 in the Tarai/Madhesh region following the promulgation of the Interim Constitution that left the incorporation of federalism and proportional representation in the Constitution. The most excluded community in Nepal- Madheshi community, seemed to have angered against state-sponsored discrimination and exclusion that have been occurred for centuries. Since Madheshis were treated as non-Nepali, though the history of Nepal’s Tarai/Madhesh has been ancient. In the beginning, this movement was against Maoist, but later, it went against the state's prejudices and discriminations. It extended across the Tarai/Madhesh region of Nepal for a month. The movement was spontaneous to a large extent. A researcher himself is a witness to the movement. Key Informant Interviews with participants, including politicians, journalists, and activists, have mainly carried out for the study. This movement ensured Madheshi identity first. Secondly, the number of electoral constituencies was increased as it reached 120 in Tarai/Madhesh while it was 80 only. As a result, Madheshi representation in the Constitution Assembly reached 35 %, while it was 20% only. The main thing that this movement played a major role in ensuring the federalism as a political system in Nepal.

Keywords: dignity, exclusion, federalism, inclusion, Madhesh movement, nation-building

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2888 Political Discourse and Linguistic Manipulation in Nigerian Politics

Authors: Kunle Oparinde, Ernestina Maleshoane Rapeane-Mathonsi, Gift Mheta

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Using Critical Discourse Analysis (CDA) and Multimodal Discourse Analysis (MDA), the research seeks to deconstruct politically-motivated discourse as observed from Nigerian politics. This is intended to be achieved by analysing linguistic (mis)representation and manipulation in Nigerian political settings, drawing from instances of language use as observed from different political campaigns. Since language in itself is generally meaningless without context, it is therefore paramount to analyse the (mis)representation and manipulation in Nigerian political sceneries within their contextual basis. The study focuses on political language used by Nigerian politicians emanating from printed and social media forms such as posters, pamphlets, speeches, billboards, and internet sources purposely selected across Nigeria. The research further aims at investigating the discursive strategies used by politicians to gain more audience, and, as a result, shape opinions that result in votes. The study employs a qualitative approach. Two parties are intentionally selected because they have been essentially strong at the national level namely: All Progressive Congress (APC) and the People’s Democratic Party (PDP). The study finds out that politicians in Nigeria, as in many parts of the world, use language to manipulate the electorate. Comprehensive discussion of these instances of political manipulation remains the thrust of this paper.

Keywords: communication, discourse, manipulation, misrepresentation

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2887 The Influence of Concreteness on English Compound Noun Processing: Modulation of Constituent Transparency

Authors: Turgut Coskun

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'Concreteness effect' refers to faster processing of concrete words and 'compound facilitation' refers to faster response to compounds. In this study, our main goal was to investigate the interaction between compound facilitation and concreteness effect. The latter might modulate compound processing basing on constituents’ transparency patterns. To evaluate these, we created lists for compound and monomorphemic words, sub-categorized them into concrete and abstract words, and further sub-categorized them basing on their transparency. The transparency conditions were opaque-opaque (OO), transparent-opaque (TO), and transparent-transparent (TT). We used RT data from English Lexicon Project (ELP) for our comparisons. The results showed the importance of concreteness factor (facilitation) in both compound and monomorphemic processing. Important for our present concern, separate concrete and abstract compound analyses revealed different patterns for OO, TO, and TT compounds. Concrete TT and TO conditions were processed faster than Concrete OO, Abstract OO and Abstract TT compounds, however, they weren’t processed faster than Abstract TO compounds. These results may reflect on different representation patterns of concrete and abstract compounds.

Keywords: abstract word, compound representation, concrete word, constituent transparency, processing speed

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2886 Restrictedly-Regular Map Representation of n-Dimensional Abstract Polytopes

Authors: Antonio Breda d’Azevedo

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Regularity has often been present in the form of regular polyhedra or tessellations; classical examples are the nine regular polyhedra consisting of the five Platonic solids (regular convex polyhedra) and the four Kleper-Poinsot polyhedra. These polytopes can be seen as regular maps. Maps are cellular embeddings of graphs (with possibly multiple edges, loops or dangling edges) on compact connected (closed) surfaces with or without boundary. The n-dimensional abstract polytopes, particularly the regular ones, have gained popularity over recent years. The main focus of research has been their symmetries and regularity. Planification of polyhedra helps its spatial construction, yet it destroys its symmetries. To our knowledge there is no “planification” for n-dimensional polytopes. However we show that it is possible to make a “surfacification” of the n-dimensional polytope, that is, it is possible to construct a restrictedly-marked map representation of the abstract polytope on some surface that describes its combinatorial structures as well as all of its symmetries. We also show that there are infinitely many ways to do this; yet there is one that is more natural that describes reflections on the sides ((n−1)-faces) of n-simplices with reflections on the sides of n-polygons. We illustrate this construction with the 4-tetrahedron (a regular 4-polytope with automorphism group of size 120) and the 4-cube (a regular 4-polytope with automorphism group of size 384).

Keywords: abstract polytope, automorphism group, N-simplicies, symmetry

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2885 Conflict Resolution in Fuzzy Rule Base Systems Using Temporal Modalities Inference

Authors: Nasser S. Shebka

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Fuzzy logic is used in complex adaptive systems where classical tools of representing knowledge are unproductive. Nevertheless, the incorporation of fuzzy logic, as it’s the case with all artificial intelligence tools, raised some inconsistencies and limitations in dealing with increased complexity systems and rules that apply to real-life situations and hinders the ability of the inference process of such systems, but it also faces some inconsistencies between inferences generated fuzzy rules of complex or imprecise knowledge-based systems. The use of fuzzy logic enhanced the capability of knowledge representation in such applications that requires fuzzy representation of truth values or similar multi-value constant parameters derived from multi-valued logic, which set the basis for the three t-norms and their based connectives which are actually continuous functions and any other continuous t-norm can be described as an ordinal sum of these three basic ones. However, some of the attempts to solve this dilemma were an alteration to fuzzy logic by means of non-monotonic logic, which is used to deal with the defeasible inference of expert systems reasoning, for example, to allow for inference retraction upon additional data. However, even the introduction of non-monotonic fuzzy reasoning faces a major issue of conflict resolution for which many principles were introduced, such as; the specificity principle and the weakest link principle. The aim of our work is to improve the logical representation and functional modelling of AI systems by presenting a method of resolving existing and potential rule conflicts by representing temporal modalities within defeasible inference rule-based systems. Our paper investigates the possibility of resolving fuzzy rules conflict in a non-monotonic fuzzy reasoning-based system by introducing temporal modalities and Kripke's general weak modal logic operators in order to expand its knowledge representation capabilities by means of flexibility in classifying newly generated rules, and hence, resolving potential conflicts between these fuzzy rules. We were able to address the aforementioned problem of our investigation by restructuring the inference process of the fuzzy rule-based system. This is achieved by using time-branching temporal logic in combination with restricted first-order logic quantifiers, as well as propositional logic to represent classical temporal modality operators. The resulting findings not only enhance the flexibility of complex rule-base systems inference process but contributes to the fundamental methods of building rule bases in such a manner that will allow for a wider range of applicable real-life situations derived from a quantitative and qualitative knowledge representational perspective.

Keywords: fuzzy rule-based systems, fuzzy tense inference, intelligent systems, temporal modalities

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2884 Potential of Grass Silage as a Source of Nutrients in Poultry Production

Authors: Hamim Abbas, Jean Luc-Hornick, Isabelle Dufrasne

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Feed costs constitute over 60% of total expenses in organic layer poultry production, with feed protein supply being a significant concern. Alfalfa-based dehydrated silage pellets are mainly diets composed of leaves (ABSP), which are non-conventional protein sources that could enhance profits by reducing feed costs and ensuring consistent availability. This experiment studied the effects on the performances of Novogen Brown light layers of a commercial control diet replaced with 10% ABSP. After a 21-day trial, this diet (ABSP) has improved the laying rate, yolk color of eggs, feed conversion rate, ω−3 (PUFAs) and ω−6/ω−3 ratio (P<0.05) while the body weight and egg weight were degraded with the substitution of the ABSP in the diet(P>0.05). The laying rate showed a tendency to increase (P=0.06). These findings suggest that ABSP can replace at least 10% of the feed in organic layer diets without compromising production parameters negatively.

Keywords: alfalfa, silage, pellet, organic layers

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2883 Admission C-Reactive Protein Serum Levels and In-Hospital Mortality in the Elderly Admitted to the Acute Geriatrics Department

Authors: Anjelika Kremer, Irina Nachimov, Dan Justo

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Background: C-reactive protein (CRP) serum levels are commonly measured in hospitalized patients. Elevated admission CRP serum levels and in-hospital mortality has been seldom studied in the general population of elderly patients admitted to the acute Geriatrics department. Methods: A retrospective cross-sectional study was conducted at a tertiary medical center. Included were all elderly patients (age 65 years or more) admitted to a single acute Geriatrics department from the emergency room between April 2014 and January 2015. CRP serum levels were measured routinely in all patients upon the first 24 hours of admission. A logistic regression analysis was used to study if admission CRP serum levels were associated with in-hospital mortality independent of age, gender, functional status, and co-morbidities. Results: Overall, 498 elderly patients were included in the analysis: 306 (61.4%) female patients and 192 (38.6%) male patients. The mean age was 84.8±7.0 years (median: 85 years; IQR: 80-90 years). The mean admission CRP serum levels was 43.2±67.1 mg/l (median: 13.1 mg/l; IQR: 2.8-51.7 mg/l). Overall, 33 (6.6%) elderly patients died during the hospitalization. A logistic regression analysis showed that in-hospital mortality was independently associated with history of stroke (p < 0.0001), heart failure (p < 0.0001), and admission CRP serum levels (p < 0.0001) – and to a lesser extent with age (p = 0.042), collagen vascular disease (p=0.011), and recent venous thromboembolism (p=0.037). Receiver operating characteristic (ROC) curve showed that admission CRP serum levels predict in-hospital mortality fairly with an area under the curve (AUC) of 0.694 (p < 0.0001). Cut-off value with maximal sensitivity and specificity was 19.7 mg/L. Conclusions: Admission CRP serum levels may be used to predict in-hospital mortality in the general population of elderly patients admitted to the acute Geriatrics department.

Keywords: c-reactive protein, elderly, mortality, prediction

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2882 Biodistribution of Fluorescence-Labelled Epidermal Growth Factor Protein from Slow Release Nanozolid Depots in Mouse

Authors: Stefan Gruden, Charlott Brunmark, Bo Holmqvist, Erwin D. Brenndorfer, Martin Johansson, Jian Liu, Ying Zhao, Niklas Axen, Moustapha Hassan

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Aim: The study was designed to evaluate the ability of the calcium sulfate-based NanoZolid® drug delivery technology to locally release the epidermal growth factor (EGF) protein while maintaining its biological activity. Methods: NanoZolid-formulated EGF protein labelled with a near-infrared dye (EGF-NIR) depots or EGF-NIR dissolved in PBS were injected subcutaneously into mice bearing EGF receptor (EGFR) positive human A549 lung cancer tumors inoculated subcutaneously. The release and biodistribution of the EGF-NIR were investigated in vivo longitudinally up to 96 hours post-administration, utilizing whole-body fluorescence imaging. In order to confirm the in vivo findings, histological analysis of tumor cryosections was performed to investigate EGF-NIR fluorescent signal and EGFR expression level by immunofluorescence labelling. Results: The in vivo fluorescence imaging showed a controlled release profile of the EGF-NIR loaded in the NanoZolid depots compared to free EGF-NIR. Histological analysis of the tumors further demonstrated a prevailing distribution of EGF-NIR in regions with high levels of EGFR expression. Conclusion: Calcium sulfate based depots can be used to formulate EGF while maintaining its biological activity, e.g., receptor binding capability. This may have good clinical potential for local delivery of biomolecules to enhance treatment efficacy and minimize systemic adverse effects.

Keywords: bioresorbable, calcium sulfate, controlled release, NanoZolid

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2881 Principles to Design Urbanism in Cinema; An Aesthetic Study on Identity and Representation of a City in a Movie

Authors: Dorsa Moayedi

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‘The Cities’ and Cinema have a history going as far back as silent films; however, the standards of picturing a city in a film are somewhat vague. ‘Genius Loci’ of a city can be easily described with parameters that architects have detected; nevertheless, the genius loci of an ‘urban movie’ is untouched. Cities have been among the provocative matters that pushed filmmakers to ponder upon them and to picture them along with their urban identity thoroughly in their artworks, though the impacts of the urban life on the plot and characters is neglected, and so a city in a movie is usually restricted to ‘the place where the story happens’. Cities and urban life are among those that are in constant change and ongoing expansion; therefore, they are always fresh and ready to challenge people with their existence. Thus, the relationship between the city and cinema is metamorphic, though it could be defined and explored. The dominant research on the idea of urbanism has been conducted by outstanding scholars of architecture, like Christian Norberg-Schulz, and the studies on Cinema have been done by theorists of cinema, like Christian Metz, who have mastered defining their own realm; still, the idea to mingle the domains to reach a unified theory which could be applied to ‘urban movies’ is barely worked on. In this research, we have sought mutual grounds to discuss ‘urbanism in cinema,’ the grounds that cinema could benefit from and get to a more accurate audio-visual representation of a city, in accordance with the ideas of Christopher Alexander and the term he coined ‘The Timeless Way of Building.’ We concentrate on movies that are dependent on urban life, mainly those that possess the names of cities, like ‘Nashville (1975), Manhattan (1979), Fargo (1996), Midnight in Paris (2011) or Roma (2018), according to the ideas of urban design and narratives of cinema. Contrary to what has often been assumed, cinema and architecture could be defined in line with similar parameters, and architectural terms could be applied to the research done on movies. Our findings indicate that the theories of Christopher Alexander can best fit the paradigm to study an ‘Urban Movie’, definitions of a timeless building, elaborate on the characteristics of a design that could be applied to definitions of an urban movie, and set a prototype for further filmmaking regarding the urban life.

Keywords: city, urbanism, urban movies, identity, representation

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2880 Event Data Representation Based on Time Stamp for Pedestrian Detection

Authors: Yuta Nakano, Kozo Kajiwara, Atsushi Hori, Takeshi Fujita

Abstract:

In association with the wave of electric vehicles (EV), low energy consumption systems have become more and more important. One of the key technologies to realize low energy consumption is a dynamic vision sensor (DVS), or we can call it an event sensor, neuromorphic vision sensor and so on. This sensor has several features, such as high temporal resolution, which can achieve 1 Mframe/s, and a high dynamic range (120 DB). However, the point that can contribute to low energy consumption the most is its sparsity; to be more specific, this sensor only captures the pixels that have intensity change. In other words, there is no signal in the area that does not have any intensity change. That is to say, this sensor is more energy efficient than conventional sensors such as RGB cameras because we can remove redundant data. On the other side of the advantages, it is difficult to handle the data because the data format is completely different from RGB image; for example, acquired signals are asynchronous and sparse, and each signal is composed of x-y coordinate, polarity (two values: +1 or -1) and time stamp, it does not include intensity such as RGB values. Therefore, as we cannot use existing algorithms straightforwardly, we have to design a new processing algorithm to cope with DVS data. In order to solve difficulties caused by data format differences, most of the prior arts make a frame data and feed it to deep learning such as Convolutional Neural Networks (CNN) for object detection and recognition purposes. However, even though we can feed the data, it is still difficult to achieve good performance due to a lack of intensity information. Although polarity is often used as intensity instead of RGB pixel value, it is apparent that polarity information is not rich enough. Considering this context, we proposed to use the timestamp information as a data representation that is fed to deep learning. Concretely, at first, we also make frame data divided by a certain time period, then give intensity value in response to the timestamp in each frame; for example, a high value is given on a recent signal. We expected that this data representation could capture the features, especially of moving objects, because timestamp represents the movement direction and speed. By using this proposal method, we made our own dataset by DVS fixed on a parked car to develop an application for a surveillance system that can detect persons around the car. We think DVS is one of the ideal sensors for surveillance purposes because this sensor can run for a long time with low energy consumption in a NOT dynamic situation. For comparison purposes, we reproduced state of the art method as a benchmark, which makes frames the same as us and feeds polarity information to CNN. Then, we measured the object detection performances of the benchmark and ours on the same dataset. As a result, our method achieved a maximum of 7 points greater than the benchmark in the F1 score.

Keywords: event camera, dynamic vision sensor, deep learning, data representation, object recognition, low energy consumption

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2879 A Secreted Protein Can Attenuate High Fat Diet Induced Obesity and Metabolic Syndrome in Mice

Authors: Abdul Soofi, Katherine Wolf, Egon Ranghini, Gregory Dressler

Abstract:

Obesity and its associated complications, such as insulin resistance and non-alcoholic fatty liver disease, are reaching epidemic proportions. In mice, the TGF-β superfamily is implicated in the regulation of white and brown adipose tissues differentiation. The Kielin/Chordin-like Protein (KCP) is a secreted regulator of the TGF-β superfamily pathways that can inhibit both TGF-β and Activin signals while enhancing the Bone Morphogenetic protein (BMP) signaling. However, the effects of KCP on metabolism and obesity have not been studied in animal models. Thus, we examined the effects of KCP loss or gain of function in mice that were maintained on either a regular or a high fat diet. Loss of KCP sensitized mice to obesity and associated complications such as hepatic steatosis and glucose intolerance. In contrast, transgenic mice that expressed KCP in the kidney, liver and adipose tissues were resistant to developing high fat diet induced obesity and had significantly reduced white adipose tissue. KCP over-expression was able to shift the pattern of Smad signaling in vivo, to increase the levels of P-Smad1 and decrease P-Smad3, resulting in resistance to high fat diet induced hepatic steatosis and glucose intolerance. In aging mice, loss of KCP promoted liver pathology even when mice were fed a normal diet. The data demonstrate that shifting the TGF-β superfamily signaling with a secreted inhibitor or enhancer can alter the physiology of adipose tissue to reduce obesity and can inhibit the initiation and progression of hepatic steatosis to significantly reduce the effects of high fat diet induced metabolic disease.

Keywords: adipose tissue, KCP, obesity, TGF-β, BMP, hepatic steatosis, metabolic syndrome

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2878 Utilization Of Guar Gum As Functional Fat Replacer In Goshtaba, A Traditional Indian Meat Product

Authors: Sajad A. Rather, F. A. Masoodi, Rehana Akhter, S. M. Wani, Adil Gani

Abstract:

Modern trend towards convenience foods has resulted in increased production and consumption of restructured meat products and are of great importance to the meat industry. In meat products fat plays an important role in cooking properties, texture & sensory scores, however, high fat contents in particular animal fats provide high amounts of saturated fatty acids and cholesterol and are associated with several types of non communicable diseases such as obesity, hypertension and coronary heart diseases. Thus, fat reduction has generally been seen as an important strategy to produce healthier meat products. This study examined the effects of reducing fat level from 20% to 10% and substituting mutton back fat with guar gum (0.5%, 1% & 1.5%) on cooking properties, proximate composition, lipid and protein oxidation, texture, microstructure and sensory characteristics of goshtaba- a traditional meat product of J & K, India were investigated and compared with high fat counterparts. Reduced- fat goshtaba samples containing guar gum had significantly (p ≤ 0.05) higher yield, less shrinkage, more moisture retention and more protein content than the control sample. TBARs and protein oxidation (carbonyl content) values of the control was significantly (p ≤ 0.05) higher than reduced fat goshtaba samples and showed a positive correlation between lipid and protein oxidation. Hardness, gumminess & chewiness of the control (20%) were significantly higher than reduced fat goshtaba samples. Microstructural differences were significant (p ≤ 0.05) between control and treated samples due to an increased moisture content in the reduced fat samples. Sensory evaluation showed significant (p ≤ 0.05) reduction in texture, flavour and overall acceptability scores of treatment products; however the scores for 0.5% and 1% treated samples were in the range of acceptability. Guar gum may also be used as a source of soluble dietary fibre in food products and a number of clinical studies have shown a reduction in postprandial glycemia and insulinemia on consumption of guar gum, with the mechanism being attributed to an increased transit time in the stomach and small intestine, which may have been due to the viscosity of the meal hindering the access of glucose to the epithelium.

Keywords: goshtaba, guar gum, traditional, fat reduction, acceptability

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2877 Violence Detection and Tracking on Moving Surveillance Video Using Machine Learning Approach

Authors: Abe Degale D., Cheng Jian

Abstract:

When creating automated video surveillance systems, violent action recognition is crucial. In recent years, hand-crafted feature detectors have been the primary method for achieving violence detection, such as the recognition of fighting activity. Researchers have also looked into learning-based representational models. On benchmark datasets created especially for the detection of violent sequences in sports and movies, these methods produced good accuracy results. The Hockey dataset's videos with surveillance camera motion present challenges for these algorithms for learning discriminating features. Image recognition and human activity detection challenges have shown success with deep representation-based methods. For the purpose of detecting violent images and identifying aggressive human behaviours, this research suggested a deep representation-based model using the transfer learning idea. The results show that the suggested approach outperforms state-of-the-art accuracy levels by learning the most discriminating features, attaining 99.34% and 99.98% accuracy levels on the Hockey and Movies datasets, respectively.

Keywords: violence detection, faster RCNN, transfer learning and, surveillance video

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2876 Reconstruction of Alveolar Bone Defects Using Bone Morphogenetic Protein 2 Mediated Rabbit Dental Pulp Stem Cells Seeded on Nano-Hydroxyapatite/Collagen/Poly(L-Lactide)

Authors: Ling-Ling E., Hong-Chen Liu, Dong-Sheng Wang, Fang Su, Xia Wu, Zhan-Ping Shi, Yan Lv, Jia-Zhu Wang

Abstract:

Objective: The objective of the present study is to evaluate the capacity of a tissue-engineered bone complex of recombinant human bone morphogenetic protein 2 (rhBMP-2) mediated dental pulp stem cells (DPSCs) and nano-hydroxyapatite/collagen/poly(L-lactide)(nHAC/PLA) to reconstruct critical-size alveolar bone defects in New Zealand rabbit. Methods: Autologous DPSCs were isolated from rabbit dental pulp tissue and expanded ex vivo to enrich DPSCs numbers, and then their attachment and differentiation capability were evaluated when cultured on the culture plate or nHAC/PLA. The alveolar bone defects were treated with nHAC/PLA, nHAC/PLA+rhBMP-2, nHAC/PLA+DPSCs, nHAC/PLA+DPSCs+rhBMP-2, and autogenous bone (AB) obtained from iliac bone or were left untreated as a control. X-ray and a polychrome sequential fluorescent labeling were performed post-operatively and the animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. Results: Our results showed that DPSCs expressed STRO-1 and vementin, and favoured osteogenesis and adipogenesis in conditioned media. DPSCs attached and spread well, and retained their osteogenic phenotypes on nHAC/PLA. The rhBMP-2 could significantly increase protein content, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, and mineral formation of DPSCs cultured on nHAC/PLA. The X-ray graph, the fluorescent, histological observation and histomorphometric analysis showed that the nHAC/PLA+DPSCs+rhBMP-2 tissue-engineered bone complex had an earlier mineralization and more bone formation inside the scaffold than nHAC/PLA, nHAC/PLA+rhBMP-2 and nHAC/PLA+DPSCs, or even autologous bone. Implanted DPSCs contribution to new bone were detected through transfected eGFP genes. Conclutions: Our findings indicated that stem cells existed in adult rabbit dental pulp tissue. The rhBMP-2 promoted osteogenic capability of DPSCs as a potential cell source for periodontal bone regeneration. The nHAC/PLA could serve as a good scaffold for autologous DPSCs seeding, proliferation and differentiation. The tissue-engineered bone complex with nHAC/PLA, rhBMP-2, and autologous DPSCs might be a better alternative to autologous bone for the clinical reconstruction of periodontal bone defects.

Keywords: nano-hydroxyapatite/collagen/poly (L-lactide), dental pulp stem cell, recombinant human bone morphogenetic protein, bone tissue engineering, alveolar bone

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2875 Septin 11, Cytoskeletal Protein Involved in the Regulation of Lipid Metabolism in Adipocytes

Authors: Natalia Moreno-Castellanos, Amaia Rodriguez, Gema Frühbeck

Abstract:

Introduction: In adipocytes, the cytoskeleton undergoes important expression and distribution in adipocytes rearrangements during adipogenesis and in obesity. Indeed, a role for these proteins in the regulation of adipocyte differentiation and response to insulin has been demonstrated. Recently, septins have been considered as new components of the cytoskeletal network that interact with other cytoskeletal elements (actin and tubulin) profoundly modifying their dynamics. However, these proteins have not been characterized as yet in adipose tissue. In this work, were examined the cellular, molecular and functional features of a member of this family, septin 11 (SEPT11), in adipocytes and evaluated the impact of obesity on the expression of this protein in human adipose tissue. Methods: Adipose gene and protein expression levels of SEPT11 were analysed in human samples. SEPT11 distribution was evaluated by immunocytochemistry, electronic microscopy, and subcellular fractionation techniques. GST-pull down, immunoprecipitation and a Yeast-Two Hybrid (Y2H) screening were used to identify the SEPT11 interactome. Gene silencing was employed to assess the role of SEPT11 in the regulation of insulin signaling and lipid metabolism in adipocytes. Results: SEPT11 is expressed in human adipocytes, and its levels increased in both omental and subcutaneous adipose tissue in obesity, with SEPT11 mRNA content positively correlating with parameters of insulin resistance in subcutaneous fat. In non-stimulated adipocytes, SEPT11 immunoreactivity showed a ring-like distribution at the cell surface and associated to caveolae. Biochemical analyses showed that SEPT11 interacted with the main component of caveolae, caveolin-1 (CAV1) as well as with the fatty acid-binding protein, FABP5. Notably, the three proteins redistributed and co-localized at the surface of lipid droplets upon exposure of adipocytes to oleate. In this line, SEPT11 silencing in 3T3-L1 adipocytes impaired insulin signaling and decreased insulin-induced lipogenesis. Conclusions: Those findings demonstrate that SEPT11 is a novel component of the adipocyte cytoskeleton that plays an important role in the regulation of lipid traffic, metabolism and can thus represent a potential biomarker of insulin resistance in obesity in adipocytes through its interaction with both CAV1 and FABP5.

Keywords: caveolae, lipid metabolism, obesity, septins

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2874 Augmented Reality Using Cuboid Tracking as a Support for Early Stages of Architectural Design

Authors: Larissa Negris de Souza, Ana Regina Mizrahy Cuperschmid, Daniel de Carvalho Moreira

Abstract:

Augmented Reality (AR) alters the elaboration of the architectural project, which relates to project cognition: representation, visualization, and perception of information. Understanding these features from the earliest stages of the design can facilitate the study of relationships, zoning, and overall dimensions of the forms. This paper’s goal was to explore a new approach for information visualization during the early stages of architectural design using Augmented Reality (AR). A three-dimensional marker inspired by the Rubik’s Cube was developed, and its performance, evaluated. This investigation interwovens the acquired knowledge of traditional briefing methods and contemporary technology. We considered the concept of patterns (Alexander et al. 1977) to outline geometric forms and associations using visual programming. The Design Science Research was applied to develop the study. An SDK was used in a game engine to generate the AR app. The tool's functionality was assessed by verifying the readability and precision of the reconfigurable 3D marker. The results indicated an inconsistent response. To use AR in the early stages of architectural design the system must provide consistent information and appropriate feedback. Nevertheless, we conclude that our framework sets the ground for looking deep into AR tools for briefing design.

Keywords: augmented reality, cuboid marker, early design stages, graphic representation, patterns

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2873 Impact of Heavy Metal Toxicity on Metabolic Changes in the Diazotrophic Cyanobacterium Anabaena PCC 7120

Authors: Rishi Saxena

Abstract:

Cyanobacteria is a photosynthetic prokaryote, and these obtain their energy through photosynthesis. In this paper, we studied the effect of iron on metabolic changes in the diazotrophic cyanobacterium Anabaena PCC 7120. Nowadays, metal contamination due to natural and anthropogenic sources is a global environment concern. Iron induced changes in growth, N2-fixation, CO2 fixation and photosynthetic activity were studied in a diazotrophic cyanobacterium Anabaena PCC 7120. Iron at 50 uM concentration supported the maximum growth, heterocyst frequency, CO2 fixation, photosystem I (PS I), photosystem II (PS II) and nitrogenase activities in the organism. Higher concentration of iron inhibited these processes. Chl a and PS II activities were more sensitive to iron than the protein and PS I activity. Here, it is also mentioned that heavy metal induced altered macromolecules metabolism and changes in the central dogma of life (DNA→ mRNA → Protein). And also recent advances have been made in understanding heavy metal-cyanobacteria interaction and their application for metal detoxification.

Keywords: cyanobacterium anabaena 7120, nitrogen fixation, photosystem I (PS I), photosystem II (PS II)

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2872 A Feminist Historical Institutional Approach and Gender Participation in Queensland Politics

Authors: Liz van Acker, Linda Colley

Abstract:

Political processes are shaped by the gendered culture of parliaments. This paper examines how the institution of parliament has been affected by the changing number of women in politics. In order to understand how and why gender change occurs, the paper employs a feminist historical institutionalism approach. It argues that while it is difficult to change the gendered nature of political institutions, it is possible, from a gender perspective, to understand the processes of change both formally and informally. Increasing women’s representation has been a slow process which has not occurred without political struggles. A broadly defined ‘feminist historical institutionalism’ has critiqued existing approaches to institutions and combined historical institutional analysis with tools of gender to enhance our understanding of institutional processes and change. The paper examines the gendered rules, norms, and practices that influence institutional design choices and processes. Institutions such as Parliament often are able to adjust to women’s entry and absorb them without too much interruption. Exploring the hidden aspects to informal institutions involves identifying unspoken and accepted norms that may guide decision-making – exposing and questioning the gender status quo. This paper examines the representation of women in the Queensland Parliament, Australia. It places the Queensland experience in historical context, as well as in the national and international context. The study is interesting, given that its gender representation has rocketed from one of the worst performing states in 2012 to one of the best performing in 2015 with further improvements in 2017. The state currently has a re-elected female Premier, a female Deputy Premier and a female-dominated cabinet – in fact, Queensland was the first ministry in Australia to have a majority of women in its Cabinet. However, it is unnecessary to dig far below these headlines to see that this is uncharacteristic of its history: progress towards this current position has been slow and patchy. The paper finds that matters such as the glass ceiling and the use of quotas explain women’s recent success in Queensland politics.

Keywords: feminist historical institutional approach, glass ceiling, quotas, women’s participation in politics

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2871 Effect pH on Chemical and Physical Properties of Iranian Fetta Cheese

Authors: M. Dezyani, R. Ezzati, H. Mirzaei

Abstract:

The objectives of this study were to determine the effect of pH on chemical, structural, and functional properties of Fetta cheese, and to relate changes in structure to changes in cheese unctionality. Fetta cheese was obtained from a cheese-production facility and stored at 4°C. Ten days after manufacture, the cheese was cut into blocks that were vacuum-packaged and stored for 4 d at 4°C. Cheese blocks were then high-pressure injected one, three, or five times with a 20% (wt/wt) glucono-δ-lactone solution. Successive injections were performed 24 h apart. Cheese blocks were then analyzed after 40 d of storage at 4°C. Acidulant injection decreased cheese pH from 5.3 in the uninjected cheese to 4.7 after five injections. Decreased pH increased the content of soluble calcium and slightly decreased the total calcium content of cheese. At the highest level, injection of acidulant promoted syneresis. Thus, after five injections, the moisture content of cheese decreased from 34 to 31%, which esulted in decreased cheese weight. Lowered cheese pH, 4.7 compared with 5.3, also resulted in contraction of the protein matrix. Acidulant injection decreased cheese hardness and cohesiveness, and the cheese became more crumbly.

Keywords: calcium, high-pressure injection, protein matrix, syneresis

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2870 Transcriptomic and Translational Regulation of Peroxisome Proliferator-Activated Receptors after Different Feedings in Salmon

Authors: Mahsa Jalili, Essa Ehsan Khan, Signe Dille Lovmo, Augustine Akruwe, Egil Lien, Rolf Erik Olsen, Trygve Sigholt, Atle Magnus Bones

Abstract:

Data from the Norwegian Directorate of Fisheries reported that >1.2 million tons of Atlantic salmon were produced in Norway aquaculture industry in 2016. Peroxisome proliferator-activated receptors (PPARs) are one of the key transcription factor families that respond to nutritional ligands. Recent studies have shown the connection between PPARs with lipid and carbohydrate metabolism in aquaculture. To our knowledge, there is no published data about the effects of krill meal, soybean meal, Bactocell ® and butyrate feedings compared to control group on PPARs gene and protein expressions in Atlantic salmon. Fish, 1year +postsmolt, average weight 250 gram were cultured for 12 weeks after acclimatization by control commercial feeding in 2 weeks after hatchery. Water oxygen rate, salinity, and temperature were monitored every second day. At the end of the trial, fish were taken from tanks randomly, and four replicates per group were collected and stored in -80 freezers until analysis. Total RNA extracted from posterior part of dorsal fin muscle tissues and Nanodrop and Bioanalyzer was used to check the quality of RNA. Gene expression of PPAR α, β and γ were determined by RT-PCR. The expression of genes of interest was measured relative to control group after normalization to three reference genes. Total protein concentration was calculated by Bradford method, and protein expression was determined with primary PPARγ antibody by western blot. All data were analyzed by ANOVA followed by Benjamini-Hochberg and Bonferroni tests. Probability values <0.05 considered significant. Bactocell® and butyrate groups showed significantly lower PPARα expression. PPARβ and γ were not significantly different among groups. PPARγ mRNA expression was approximately consistent with protein expression pattern, except than butyrate group showed lower mRNA level. The order of PPARγ expression was Bactocell® > soy meal > butyrate > krill meal > control respectively. PPARβ gene expression decreased more in soy meal > butyrate > krill meal > Bactocell® > control groups respectively. In conclusion, the increased expression of PPARγ and α is proposed to represent a reduction tendency of lipid storage in fish fed by Bactocell®, butyrate, soy and krill meal.

Keywords: aquaculture, blotting western, gene expression, krill protein extract, prebiotics, probiotics, Salmo salar

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2869 Oxidative Damage to Lipids, Proteins, and DNA during Differentiation of Mesenchymal Stem Cells Derived from Umbilical Cord into Biologically Active Hepatocytes

Authors: Abdolamir Allameh, Shahnaz Esmaeili, Mina Allameh, Safoura Khajeniazi

Abstract:

Stem cells with therapeutic applications can be isolated from human placenta/umblical cord blood (UCB) as well as the cord tissue (UC). Stem cells in culture are vulnerable to oxidative stress, particularly when subjected to differentiation process. The aim of this study was to examine the chnages in the rate of oxidation that occurs to cellular macromolecules during hepatic differentiation of mononuclear cells (MSCs). In addition, the impact of the hepatic differentiation process of MSC on cellular and biological activity of the cells will be undertaken. For this purpose, first mononuclear cells (MNCs) were isolated from human UCB which was obtained from a healthy full-term infant. The cells were cultured at a density of 3×10⁵ cells/cm² in DMEM- low-glucose culture media supplemented with 20% FBS, 2 mM L-glutamine, 100 μg/ml streptomycin and 100 U/ml penicillin. Cell cultures were then incubated at 37°C in a humidified 5% CO₂ incubator. After removing non-adherent cells by replacing culture medium, fibroblast-like adherent cells were resuspended in 0.25% trypsin-EDTA and plated in 25 cm² flasks (1×10⁴/ml). Characterization of the MSCs was routinely done by observing their morphology and growth curve. MSCs were subjected to a 2-step hepatocyte differentiation protocol in presence of hepatocyte growth factor (HGF), dexamethazone (DEX) and oncostatin M (OSM). The hepatocyte-like cells derived from MSCs were checked every week for 3 weeks for changes in lipid peroxidation, protein carbonyl formation and DNA oxidation i.e., 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assay. During the 3-week differentiation process of MSCs to hepatocyte-like cells we found that expression liver-specific markers such as albumin, was associated with increased levels of lipid peroxidation and protein carbonyl formation. Whereas, undifferentiated MSCs has relatively low levels of lipid peroxidation products. There was a significant increase ( p < 0.05) in lipid peroxidation products in hepatocytes on days 7, 14, and 21 of differentiation. Likewise, the level of protein carbonyls in the cells was elevated during the differentiation. The level of protein carbonyls measured in hepatocyte-like cells obtained 3 weeks after differentiation induction was estimated to be ~6 fold higher compared to cells recovered on day 7 of differentiation. On the contrary, there was a small but significant decrease in DNA damage marker (8-OH-dG) in hepatocytes recovered 3 weeks after differentiation onset. The level of 8-OHdG which was in consistent with formation of reactive oxygen species (ROS). In conclusion, this data suggest that despite the elevation in oxidation of lipid and protein molecules during hepatocyte development, the cells were normal in terms of DNA integrity, morphology, and biologically activity.

Keywords: adult stem cells, DNA integrity, free radicals, hepatic differentiation

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