Search results for: substance of expression
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2513

Search results for: substance of expression

1433 Control of Lymphatic Remodelling by miR-132

Authors: Valeria Arcucci, Musarat Ishaq, Steven A. Stacker, Greg J. Goodall, Marc G. Achen

Abstract:

Metastasis is the lethal aspect of cancer for most patients. Remodelling of lymphatic vessels associated with a tumour is a key initial step in metastasis because it facilitates the entry of cancer cells into the lymphatic vasculature and their spread to lymph nodes and distant organs. Although it is clear that vascular endothelial growth factors (VEGFs), such as VEGF-C and VEGF-D, are key drivers of lymphatic remodelling, the means by which many signaling pathways in endothelial cells are coordinately regulated to drive growth and remodelling of lymphatics in cancer is not understood. We seek to understand the broader molecular mechanisms that control cancer metastasis, and are focusing on microRNAs, which coordinately regulate signaling pathways involved in complex biological responses in health and disease. Here, using small RNA sequencing, we found that a specific microRNA, miR-132, is upregulated in expression in lymphatic endothelial cells (LECs) in response to the lymphangiogenic growth factors. Interestingly, ectopic expression of miR-132 in LECs in vitro stimulated proliferation and tube formation of these cells. Moreover, miR-132 is expressed in lymphatic vessels of a subset of human breast tumours which were previously found to express high levels of VEGF-D by immunohistochemical analysis on tumour tissue microarrays. In order to dissect the complexity of regulation by miR-132 in lymphatic biology, we performed Argonaute HITS-CLIP, which led us to identify the miR-132-mRNA interactome in LECs. We found that this microRNA in LECs is involved in the control of many different pathways mainly involved in cell proliferation and regulation of the extracellular matrix and cell-cell junctions. We are now exploring the functional significance of miR-132 targets in the biology of LECs using biochemical techniques, functional in vitro cell assays and in vivo lymphangiogenesis assays. This project will ultimately define the molecular regulation of lymphatic remodelling by miR-132, and thereby identify potential therapeutic targets for drugs designed to restrict the growth and remodelling of tumour lymphatics resulting in metastatic spread.

Keywords: argonaute HITS-CLIP, cancer, lymphatic remodelling, miR-132, VEGF

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1432 Optical Fiber Data Throughput in a Quantum Communication System

Authors: Arash Kosari, Ali Araghi

Abstract:

A mathematical model for an optical-fiber communication channel is developed which results in an expression that calculates the throughput and loss of the corresponding link. The data are assumed to be transmitted by using of separate photons with different polarizations. The derived model also shows the dependency of data throughput with length of the channel and depolarization factor. It is observed that absorption of photons affects the throughput in a more intensive way in comparison with that of depolarization. Apart from that, the probability of depolarization and the absorption of radiated photons are obtained.

Keywords: absorption, data throughput, depolarization, optical fiber

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1431 COX-2 Inhibitor NS398 Counteracts Chemoresistance to Temozolomide in T98G Glioblastoma Cell Line

Authors: Francesca Lombardi, Francesca Rosaria Augello, Benedetta Cinque, Maria Grazia Cifone, Paola Palumbo

Abstract:

Glioblastoma multiforme (GBM) is a high-grade primary brain tumor refractory to current forms of treatment. The survival benefits of patients with GBM remain unsatisfactory due to the intrinsic or acquired resistance to temozolomide (TMZ), an alkylating agent, used as the first-line chemotherapeutic drug to treat GBM patients. Its cytotoxic effect is visualized by the induction of O6-methylguanine (O6MeG) within DNA. Cyclooxygenase-2 (COX-2), an inflammation-associated enzyme, has been implicated in tumorigenesis and progression of GBM, its inhibition shows anticancer activities. In the present study, it was verified if the combination of a COX-2 selective inhibitor, NS398, with TMZ could counteract the TMZ resistance. In particular, the effect of NS398 mixed with TMZ was investigated in the GBM TMZ-resistant cell line, T98G. Cells were pretreated with NS398 (100µM, 24 hours) and then exposed to TMZ alone (200µM), NS398 alone, or both for 72 hours, after which cell growth rate and cycle phases, as well as apoptosis level, were evaluated. Coadministration of NS398 and TMZ caused a significant decrease in cell growth and a progressive increase of dead cells detected by trypan blue staining. Moreover, a significant level of apoptotic cell percentage and alteration of cell cycle phases were observed in T98G treated with TMZ-NS398 combination when compared to untreated cells or TMZ-treated cells. TMZ-resistant tumors, as GBM, express elevated levels of DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT). The mixture drastically reduced MGMT expression in the TMZ-resistant cell line T98G, known to express high levels of MGMT basically. Moreover, while TMZ alone did not influence the COX-2 protein expression, the combination successfully reduced it. In conclusion, these results demonstrated that NS398 enhanced the efficacy of TMZ through cell number reduction, apoptosis induction, and decreased MGMT levels, suggesting the ability of drug combination to reduce the chemoresistance. This drug combination deserves attention and could be considered as a promising therapeutic strategy for GBM patients.

Keywords: COX-2, COX-2 inhibitor, glioblastoma, NS398, T98G, temozolomide

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1430 Unzipping the Stress Response Genes in Moringa oleifera Lam. through Transcriptomics

Authors: Vivian A. Panes, Raymond John S. Rebong, Miel Q. Diaz

Abstract:

Moringa oleifera Lam. is known mainly for its high nutritional value and medicinal properties contributing to its popular reputation as a 'miracle plant' in the tropical climates where it usually grows. The main objective of this study is to discover the genes and gene products involved in abiotic stress-induced activity that may impact the M. oleifera Lam. mature seeds as well as their corresponding functions. In this study, RNA-sequencing and de novo transcriptome assembly were performed using two assemblers, Trinity and Oases, which produced 177,417 and 120,818 contigs respectively. These transcripts were then subjected to various bioinformatics tools such as Blast2GO, UniProt, KEGG, and COG for gene annotation and the analysis of relevant metabolic pathways. Furthermore, FPKM analysis was performed to identify gene expression levels. The sequences were filtered according to the 'response to stress' GO term since this study dealt with stress response. Clustered Orthologous Groups (COG) showed that the highest frequencies of stress response gene functions were those of cytoskeleton which make up approximately 14% and 23% of stress-related sequences under Trinity and Oases respectively, recombination, repair and replication at 11% and 14% respectively, carbohydrate transport and metabolism at 23% and 9% respectively and defense mechanisms 16% and 12% respectively. KEGG pathway analysis determined the most abundant stress-response genes in the phenylpropanoid biosynthesis at counts of 187 and 166 pathways for Oases and Trinity respectively, purine metabolism at 123 and 230 pathways, and biosynthesis of antibiotics at 105 and 102. Unique and cumulative GO term counts revealed that majority of the stress response genes belonged to the category of cellular response to stress at cumulative counts of 1,487 to 2,187 for Oases and Trinity respectively, defense response at 754 and 1,255, and response to heat at 213 and 208, response to water deprivation at 229 and 228, and oxidative stress at 508 and 488. Lastly, FPKM was used to determine the levels of expression of each stress response gene. The most upregulated gene encodes for thiamine thiazole synthase chloroplastic-like enzyme which plays a significant role in DNA damage tolerance. Data analysis implies that M. oleifera stress response genes are directed towards the effects of climate change more than other stresses indicating the potential of M. oleifera for cultivation in harsh environments because it is resistant to climate change, pathogens, and foreign invaders.

Keywords: stress response, genes, Moringa oleifera, transcriptomics

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1429 A Novel Upregulated circ_0032746 on Sponging with MIR4270 Promotes the Proliferation and Migration of Esophageal Squamous Cell Carcinoma

Authors: Sachin Mulmi Shrestha, Xin Fang, Hui Ye, Lihua Ren, Qinghua Ji, Ruihua Shi

Abstract:

Background: Esophageal squamous cell carcinoma (ESCC) is a tumor arising from esophageal epithelial cells and is one of the major disease subtype in Asian countries, including China. Esophageal cancer is the 7th highest incidence based on the 2020 data of GLOBOCAN. The pathogenesis of cancer is still not well understood as many molecular and genetic basis of esophageal carcinogenesis has yet to be clearly elucidated. Circular RNAs are RNA molecules that are formed by back-splicing covalently joined 3′- and 5′-endsrather than canonical splicing, and recent data suggest circular RNAs could sponge miRNAs and are enriched with functional miRNA binding sites. Hence, we studied the mechanism of circular RNA, its biological function, and the relationship between microRNA in the carcinogenesis of ESCC. Methods: 4 pairs of normal and esophageal cancer tissues were collected in Zhongda hospital, affiliated to Southeast University, and high-throughput RNA sequencing was done. The result revealed that circ_0032746 was upregulated, and thus we selected circ_0032746 for further study. The backsplice junction of circRNA was validated by sanger sequence, and stability was determined by RNASE R assay. The binding site of circRNA and microRNA was predicted by circinteractome,mirandaand RNAhybrid database. Furthermore, circRNA was silenced by siRNA and then by lentivirus. The regulatory axis of circ0032746/miR4270 was validated by shRNA, mimic, and inhibitor transfection. Then, in vitro experiments were performed to assess the role of circ0032746 on proliferation (CCK-8 assay and colon formation assay), migration and invasion (Transewell assay), and apoptosis of ESCC. Results: The upregulated circ0032746 was validated in 9 pairs of tissues and 5 types of cell lines by qPCR, which showed high expression and was statistically significant (P<0.005) ). Upregulated circ0032746 was silenced by shRNA, which showed significant knockdown in KYSE 30 and TE-1 cell lines expression compared to control. Nuclear and cytoplasmic mRNA fraction experiment displayed the cytoplasmic location of circ0032746. The sponging of miR4270 was validated by co-transfection of sh-circ0032746 and mimic or inhibitor. Transfection with mimic showed the decreased expression of circ_0032746, whereas inhibitor inhibited the result. In vitro experiments showed that silencing of circ_0032746 inhibited the proliferation, migration, and invasion compared to the negative control group. The apoptosis was seen higher in a knockdown group than in the control group. Furthermore, 11 common mircoRNA target mRNAs were predicted by Targetscan, MirTarbase, and miRanda database, which may further play role in the pathogenesis. Conclusion: Our results showed that novel circ_0032746 is upregulated in ESCC and plays role in itsoncogenicity. Silencing of circ_0032746 inhibits the proliferation and migration of ESCC whereas increases the apoptosis of cancer cells. Hence, circ0032746 acts as an oncogene in ESCC by sponging with miR4270 and could be a potential biomarker in the diagnosis of ESCC in the future.

Keywords: circRNA, esophageal squamous cell carcinoma, microRNA, upregulated

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1428 Affective Robots: Evaluation of Automatic Emotion Recognition Approaches on a Humanoid Robot towards Emotionally Intelligent Machines

Authors: Silvia Santano Guillén, Luigi Lo Iacono, Christian Meder

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One of the main aims of current social robotic research is to improve the robots’ abilities to interact with humans. In order to achieve an interaction similar to that among humans, robots should be able to communicate in an intuitive and natural way and appropriately interpret human affects during social interactions. Similarly to how humans are able to recognize emotions in other humans, machines are capable of extracting information from the various ways humans convey emotions—including facial expression, speech, gesture or text—and using this information for improved human computer interaction. This can be described as Affective Computing, an interdisciplinary field that expands into otherwise unrelated fields like psychology and cognitive science and involves the research and development of systems that can recognize and interpret human affects. To leverage these emotional capabilities by embedding them in humanoid robots is the foundation of the concept Affective Robots, which has the objective of making robots capable of sensing the user’s current mood and personality traits and adapt their behavior in the most appropriate manner based on that. In this paper, the emotion recognition capabilities of the humanoid robot Pepper are experimentally explored, based on the facial expressions for the so-called basic emotions, as well as how it performs in contrast to other state-of-the-art approaches with both expression databases compiled in academic environments and real subjects showing posed expressions as well as spontaneous emotional reactions. The experiments’ results show that the detection accuracy amongst the evaluated approaches differs substantially. The introduced experiments offer a general structure and approach for conducting such experimental evaluations. The paper further suggests that the most meaningful results are obtained by conducting experiments with real subjects expressing the emotions as spontaneous reactions.

Keywords: affective computing, emotion recognition, humanoid robot, human-robot-interaction (HRI), social robots

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1427 Effects of Extract from Lactuca sativa on Sleep in Pentobarbital-Induced Sleep and Caffeine-Induced Sleep Disturbance in Mice

Authors: Hae Dun Kim, Joo Hyun Jang, Geu Rim Seo, Kyung Soo Ra, Hyung Joo Suh

Abstract:

Lactuca sativa (lettuce) has been known for its medical property to relieve anxiety and nervous. This study was implemented to investigate sleep-promoting effects of the lettuce alcohol extract (LAE). Caffeine is widely used psychoactive substance known to induced wakefulness and insomnia to its consumers. In the present study, the sedative-hypnotic activity of the LAE was studied using the method of pentobarbital-induced sleep in the mouse model. The LAE was administrated to mice 30 min before the pentobarbital injection. The LAE prolonged the pentobarbital-induced sleep duration and decreased sleep latency. The effects of LAE were comparable to those of induced by diazepam. Another study was performed to examine whether LAE ameliorates caffeine-induced sleep disturbance in mice. Additionally, caffeine (10 mg/kg, p.o) delayed sleep onset and reduced sleep duration of mice. Conversely, LAE treatment (80 or 160 mg/kg, p.o), especially at 160 mg/kg, normalized the sleep disturbance induced by caffeine. LAE supplementation can counter the sleep disturbance induced by caffeine. These results suggest that LAE possess significant sedative-hypnotic activity, which supports the popular use of lettuce for treatment of insomnia and provide the basis for new drug discovery. Furthermore, these results demonstrate that the lettuce extract may be preferable for the treatment of insomnia.

Keywords: caffeine, Lactuca sativa, sleep duration, sleep latency

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1426 Laying Hens' Feed Fortified with Pectin, Xanthan Gum and Guar Gum Aims to Reduce the Cholesterol in Muscle and Egg Yolk

Authors: Novia Dwi Prabandari, Diah Ayu Asmarani

Abstract:

Soluble fiber can accelerate the metabolism of cholesterol. Pectin and gum has been used in the form of substance additive for material stabilizer and emulsifier. Pectin supplementation in laying hens can decimate the cholesterol content in egg yolk and muscle. Therefore, this laying hens’ feed is regular feed chickens enriched with soluble fiber (Pectin, Xanthan gum, and Guar gum) to produce eggs and muscle with lower cholesterol than usual.The ingredients are mixed in the ratio of concentrate 45%, corn flour 25%, soybean meal 20%, and extract of soluble fiber 10%. Once all the ingredients are mixed and then evaporated with temperature < 80 °C. Then put in the grinding machine resulting in a circular shape with holes 2-3 mm in diameter, after it dried up the water content in the feed is less than 14%. Eggs from laying hen with soluble fiber fortification feed intake will have lower cholesterol levels in eggs than regular feed. So even with the cholesterol content in the muscle, it is because chicken feed fortified with soluble fiber will accelerate the metabolism of cholesterol and cause cholesterol deposits in the chicken less. The use of this kind of laying hens feed is produce eggs with high protein content can be consumed more for people who have hypercholesterolemia.

Keywords: pectin, xanthan gum, guar gum, laying hen, cholesterol

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1425 Superiority of Bone Marrow Derived-Osteoblastic Cells (ALLOB®) over Bone Marrow Derived-Mesenchymal Stem Cells

Authors: Sandra Pietri, Helene Dubout, Sabrina Ena, Candice Hoste, Enrico Bastianelli

Abstract:

Bone Therapeutics is a bone cell therapy company addressing high unmet medical needs in the field of bone fracture repair, more specifically in non-union and delayed-union fractures where the bone repair process is impaired. The company has developed a unique allogeneic osteoblastic cell product (ALLOB®) derived from bone marrow which is currently tested in humans in the indication of delayed-union fractures. The purpose of our study was to directly compare ALLOB® vs. non-differentiated mesenchymal stem cells (MSC) for their in vitro osteogenic characteristics and their in vivo osteogenic potential in order to determine which cellular type would be the most adapted for bone fracture repair. Methods: Healthy volunteers’ bone marrow aspirates (n=6) were expended (i) into BM-MSCs using a complete MSC culture medium or (ii) into ALLOB® cells according to its manufacturing process. Cells were characterized in vitro by morphology, immunophenotype, gene expression and differentiation potential. Additionally, their osteogenic potential was assessed in vivo in the subperiosteal calvaria bone formation model in nude mice. Results: The in vitro side-by-side comparison studies showed that although ALLOB® and BM-MSC shared some common general characteristics such as the 3 minimal MSC criteria, ALLOB® expressed significantly higher levels of chondro/osteoblastic genes such as BMP2 (fold change (FC) > 100), ALPL (FC > 12), CBFA1 (FC > 3) and differentiated significantly earlier than BM-MSC toward the osteogenic lineage. Moreover the bone formation model in nude mice demonstrated that used at the same cellular concentration, ALLOB® was able to induce significantly more (160% vs.107% for control animals) bone formation than BM-MSC (118% vs. 107 % for control animals) two weeks after administration. Conclusion: Our side-by-side comparison studies demonstrated that in vitro and in vivo, ALLOB® displays superior osteogenic capacity to BM-MScs and is therefore a better candidate for the treatment of bone fractures.

Keywords: gene expression, histomorphometry, mesenchymal stem cells, osteogenic differentiation potential, preclinical

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1424 A Review of Material and Methods Used in Liner Layers in Various Landfills

Authors: S. Taghvamanesh

Abstract:

Modern landfills are highly engineered containment systems that are designed to reduce the environmental and human health impacts of solid waste (trash). In modern landfills, waste is contained by a liner system. The primary goal of the liner system is to isolate the landfill contents from the environment, thereby protecting the soil and groundwater from pollution caused by the leachate of a landfill. Landfill leachate is the most serious threat to groundwater. Therefore, it is necessary to design a system that prevents the penetration of this dangerous substance into the environment. These layers are made up of two basic elements: clay and geosynthetics. Hydraulic conductivity and flexibility are two desirable properties of these materials. There are three different types of liner systems that will be discussed in this paper. According to available data, the current article analyzed materials and methods for constructing liner layers made of distinct leachates, including various harmful components and heavy metals from all around the world. Also, this study attempted to gather data on leachates for each of the sites discussed. In conclusion, every landfill requires a specific type of liner, which depends on the type of leachate that it produces daily. It should also be emphasized that, based on available data, this article focused on the number of landfills that each country or continent possesses.

Keywords: landfill, liner layer, impervious layer, barrier layer

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1423 Decreased Tricarboxylic Acid (TCA) Cycle Staphylococcus aureus Increases Survival to Innate Immunity

Authors: Trenten Theis, Trevor Daubert, Kennedy Kluthe, Austin Nuxoll

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Staphylococcus aureus is a gram-positive bacterium responsible for an estimated 23,000 deaths in the United States and 25,000 deaths in the European Union annually. Recurring S. aureus bacteremia is associated with biofilm-mediated infections and can occur in 5 - 20% of cases, even with the use of antibiotics. Despite these infections being caused by drug-susceptible pathogens, they are surprisingly difficult to eradicate. One potential explanation for this is the presence of persister cells—a dormant type of cell that shows a high tolerance to antibiotic treatment. Recent studies have shown a connection between low intracellular ATP and persister cell formation. Specifically, this decrease in ATP, and therefore increase in persister cell formation, is due to an interrupted tricarboxylic acid (TCA) cycle. However, S. aureus persister cells’ role in pathogenesis remains unclear. Initial studies have shown that a fumC (TCA cycle gene) knockout survives challenge from aspects of the innate immune system better than wild-type S. aureus. Specifically, challenges from two antimicrobial peptides--LL-37 and hBD-3—show a log increase in survival of the fumC::N∑ strain compared to wild type S. aureus after 18 hours. Furthermore, preliminary studies show that the fumC knockout has a log more survival within a macrophage. These data lead us to hypothesize that the fumC knockout is better suited to other aspects of the innate immune system compared to wild-type S. aureus. To further investigate the mechanism for increased survival of fumC::N∑ within a macrophage, we tested bacterial growth in the presence of reactive oxygen species (ROS), reactive nitrogen species (RNS), and a low pH. Preliminary results suggest that the fumC knockout has increased growth compared to wild-type S. aureus in the presence of all three antimicrobial factors; however, no difference was observed in any single factor alone. To investigate survival within a host, a nine-day biofilm-associated catheter infection was performed on 6–8-week-old male and female C57Bl/6 mice. Although both sexes struggled to clear the infection, female mice were trending toward more frequently clearing the HG003 wild-type infection compared to the fumC::N∑ infection. One possible reason for the inability to reduce the bacterial burden is that biofilms are largely composed of persister cells. To test this hypothesis further, flow cytometry in conjunction with a persister cell marker was used to measure persister cells within a biofilm. Cap5A (a known persister cell marker) expression was found to be increased in a maturing biofilm, with the lowest levels of expression seen in immature biofilms and the highest expression exhibited by the 48-hour biofilm. Additionally, bacterial cells in a biofilm state closely resemble persister cells and exhibit reduced membrane potential compared to cells in planktonic culture, further suggesting biofilms are largely made up of persister cells. These data may provide an explanation as to why infections caused by antibiotic-susceptible strains remain difficult to treat.

Keywords: antibiotic tolerance, Staphylococcus aureus, host-pathogen interactions, microbial pathogenesis

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1422 A Study on the Role of Human Rights in the Aid Allocations of China and the United States

Authors: Shazmeen Maroof

Abstract:

The study is motivated by a desire to investigate whether there is substance to claims that, relative to traditional donors, China disregards human rights considerations when allocating overseas aid. While the stated policy of the U.S. is that consideration of potential aid recipients’ respect for human rights is mandatory, some quantitative studies have cast doubt on whether this is reflected in actual allocations. There is a lack of academic literature that formally assesses the extent to which the two countries' aid allocations differ; which is essential to test whether the criticisms of China's aid policy in comparison to that of the U.S. are justified. Using data on two standard human rights measures, 'Political Terror Scale' and 'Civil Liberties', the study analyse the two donors’ aid allocations among 125 countries over the period 2000 to 2014. The bivariate analysis demonstrated that a significant share of China’s aid flow to countries with poor human rights record. At the same time, the U.S. seems little different in providing aid to these countries. The empirical results obtained from the Fractional Logit model also provided some support to the general pessimism regarding China’s provision of aid to countries with poor human rights record, yet challenge the optimists expecting better targeted aid from the U.S. These findings are consistent with the split between humanitarian and non-humanitarian aid and in the sample of countries whose human rights record is below some threshold level.

Keywords: China's aid policy, foreign aid allocation, human rights, United States Foreign Assistance Act

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1421 A Comparative Analysis on the Impact of the Prevention and Combating of Hate Crimes and Hate Speech Bill of 2016 on the Rights to Human Dignity, Equality, and Freedom in South Africa

Authors: Tholaine Matadi

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South Africa is a democratic country with a historical record of racially-motivated marginalisation and exclusion of the majority. During the apartheid era the country was run along pieces of legislation and policies based on racial segregation. The system held a tight clamp on interracial mixing which forced people to remain in segregated areas. For example, a citizen from the Indian community could not own property in an area allocated to white people. In this way, a great majority of people were denied basic human rights. Now, there is a supreme constitution with an entrenched justiciable Bill of Rights founded on democratic values of social justice, human dignity, equality and the advancement of human rights and freedoms. The Constitution also enshrines the values of non-racialism and non-sexism. The Constitutional Court has the power to declare unconstitutional any law or conduct considered to be inconsistent with it. Now, more than two decades down the line, despite the abolition of apartheid, there is evidence that South Africa still experiences hate crimes which violate the entrenched right of vulnerable groups not to be discriminated against on the basis of race, sexual orientation, gender, national origin, occupation, or disability. To remedy this mischief parliament has responded by drafting the Prevention and Combatting of Hate Crimes and Hate Speech Bill. The Bill has been disseminated for public comment and suggestions. It is intended to combat hate crimes and hate speech based on sheer prejudice. The other purpose of the Bill is to bring South Africa in line with international human rights instruments against racism, racial discrimination, xenophobia and related expressions of intolerance identified in several international instruments. It is against this backdrop that this paper intends to analyse the impact of the Bill on the rights to human dignity, equality, and freedom. This study is significant because the Bill was highly contested and creates a huge debate. This study relies on a qualitative evaluative approach based on desktop and library research. The article recurs to primary and secondary sources. For comparative purpose, the paper compares South Africa with countries such as Australia, Canada, Kenya, Cuba, and United Kingdom which have criminalised hate crimes and hate speech. The finding from this study is that despite the Bill’s expressed positive intentions, this draft legislation is problematic for several reasons. The main reason is that it generates considerable controversy mostly because it is considered to infringe the right to freedom of expression. Though the author suggests that the Bill should not be rejected in its entirety, she notes the brutal psychological effect of hate crimes on their direct victims and the writer emphasises that a legislature can succeed to combat hate-crimes only if it provides for them as a separate stand-alone category of offences. In view of these findings, the study recommended that since hate speech clauses have a negative impact on freedom of expression it can be promulgated, subject to the legislature enacting the Prevention and Combatting of Hate-Crimes Bill as a stand-alone law which criminalises hate crimes.

Keywords: freedom of expression, hate crimes, hate speech, human dignity

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1420 Understanding Beginning Writers' Narrative Writing with a Multidimensional Assessment Approach

Authors: Huijing Wen, Daibao Guo

Abstract:

Writing is thought to be the most complex facet of language arts. Assessing writing is difficult and subjective, and there are few scientifically validated assessments exist. Research has proposed evaluating writing using a multidimensional approach, including both qualitative and quantitative measures of handwriting, spelling and prose. Given that narrative writing has historically been a staple of literacy instruction in primary grades and is one of the three major genres Common Core State Standards required students to acquire starting in kindergarten, it is essential for teachers to understand how to measure beginning writers writing development and sources of writing difficulties through narrative writing. Guided by the theoretical models of early written expression and using empirical data, this study examines ways teachers can enact a comprehensive approach to understanding beginning writer’s narrative writing through three writing rubrics developed for a Curriculum-based Measurement (CBM). The goal is to help classroom teachers structure a framework for assessing early writing in primary classrooms. Participants in this study included 380 first-grade students from 50 classrooms in 13 schools in three school districts in a Mid-Atlantic state. Three writing tests were used to assess first graders’ writing skills in relation to both transcription (i.e., handwriting fluency and spelling tests) and translational skills (i.e., a narrative prompt). First graders were asked to respond to a narrative prompt in 20 minutes. Grounded in theoretical models of earlier expression and empirical evidence of key contributors to early writing, all written samples to the narrative prompt were coded three ways for different dimensions of writing: length, quality, and genre elements. To measure the quality of the narrative writing, a traditional holistic rating rubric was developed by the researchers based on the CCSS and the general traits of good writing. Students' genre knowledge was measured by using a separate analytic rubric for narrative writing. Findings showed that first-graders had emerging and limited transcriptional and translational skills with a nascent knowledge of genre conventions. The findings of the study provided support for the Not-So-Simple View of Writing in that fluent written expression, measured by length and other important linguistic resources measured by the overall quality and genre knowledge rubrics, are fundamental in early writing development. Our study echoed previous research findings on children's narrative development. The study has practical classroom application as it informs writing instruction and assessment. It offered practical guidelines for classroom instruction by providing teachers with a better understanding of first graders' narrative writing skills and knowledge of genre conventions. Understanding students’ narrative writing provides teachers with more insights into specific strategies students might use during writing and their understanding of good narrative writing. Additionally, it is important for teachers to differentiate writing instruction given the individual differences shown by our multiple writing measures. Overall, the study shed light on beginning writers’ narrative writing, indicating the complexity of early writing development.

Keywords: writing assessment, early writing, beginning writers, transcriptional skills, translational skills, primary grades, simple view of writing, writing rubrics, curriculum-based measurement

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1419 Construction of the Large Scale Biological Networks from Microarrays

Authors: Fadhl Alakwaa

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One of the sustainable goals of the system biology is understanding gene-gene interactions. Hence, gene regulatory networks (GRN) need to be constructed for understanding the disease ontology and to reduce the cost of drug development. To construct gene regulatory from gene expression we need to overcome many challenges such as data denoising and dimensionality. In this paper, we develop an integrated system to reduce data dimension and remove the noise. The generated network from our system was validated via available interaction databases and was compared to previous methods. The result revealed the performance of our proposed method.

Keywords: gene regulatory network, biclustering, denoising, system biology

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1418 Vapochromism of 3,3’,5,5’-Tetramethylbenzidine-Tetrasilisicfluormica Intercalation Compounds with High Selectivity for Water and Acetonitrile

Authors: Reira Kinoshita, Shin'ichi Ishimaru

Abstract:

Vapochromism is a type of chromism in which the color of a substance changes when it is exposed to the vapor of volatile materials, and has been investigated for the application of chemical sensors for volatile organic compounds causing sick building syndrome and health hazards in workspaces. We synthesized intercalation compounds of 3,3',5,5'-tetramethylbenzidine (TMB), and tetrasilisicfluormica (TSFM) by the commonly used cation-exchange method with the cation ratio TMB²⁺/CEC of TSFM = 1.0, 2.0, 2.7 and 5.4 to investigate the vapochromism of these materials. The obtained samples were characterized by powder XRD, XRF, TG-DTA, N₂ adsorption, and SEM. Vapochromism was measured for each sample under a controlled atmosphere by a handy reflectance spectrometer directly from the outside of the glass sample tubes. The color was yellow for all specimens vacuum-dried at 50 °C, but it turned green under H₂O vapor exposure for the samples with TMB²⁺/CEC = 2.0, 2.7, and 5.4 and blue under acetonitrile vapor for all cation ratios. Especially the sample TMB²⁺/CEC = 2.0 showed clear chromism both for water and acetonitrile. On the other hand, no clear color change was observed for vapors of alcohols, acetone, and non-polar solvents. From these results, this material can be expected to apply for easy detection of humidity and acetonitrile vapor in the environment.

Keywords: chemical sensor, intercalation compound, tetramethylbenzidine, tetrasilisicfluormica, vapochromism, volatile organic compounds

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1417 Effects of Audiovisual Contextualization of L2 Idioms on Enhancing Students’ Comprehension and Retention

Authors: Monica Karlsson

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The positive effect of a supportive written context on comprehension and retention when faced with a previously unknown idiomatic expression is today an indisputable fact, especially if relevant clues are given in close proximity of the item in question. Also, giving learners a chance of visualizing the meaning of an idiom by offering them its source domain and/or by elaborating etymologically, i.e. providing a mental picture in addition to the spoken/written form (referred to as dual coding), seems to enhance comprehension and retention even further, especially if the idiom is of a more transparent kind. For example, by explaining that walk the plank has a maritime origin and a canary in a coal mine comes from the time when canaries were kept in cages to warn miners if gas was leaking out at which point the canaries succumbed immediately, learners’ comprehension and retention have been shown to increase. The present study aims to investigate whether contextualization of an audiovisual kind could help increase comprehension and retention of L2 idioms. 40 Swedish first-term university students studying English as part of their education to become middle-school teachers participated in the investigation, which tested 24 idioms, all of which were ascertained to be previously unknown to the informants. While half of the learners were subjected to a test in which they were asked to watch scenes from various TV programmes, each scene including one idiomatic expression in a supportive context, the remaining 20 students, as a point of reference, were only offered written contexts, though equally supportive. Immediately after these sessions, both groups were given the same idioms in a decontextualized form and asked to give their meaning. After five weeks, finally, the students were subjected to yet another decontextualized comprehension test. Furthermore, since mastery of idioms in one’s L1 appears to correlate to a great extent with a person’s ability to comprehend idioms in an L2, all the informants were also asked to take a test focusing on idioms in their L1. The result on this test is thus seen to indicate each student’s potential for understanding and memorizing various idiomatic expressions from a more general perspective. Preliminary results clearly show that audiovisual contextualization indeed has a positive effect on learners’ retention. In addition, preliminary results also show that those learners’ who were able to recall most meanings were those who had a propensity for idiom comprehension in their L1.

Keywords: English, L2, idioms, audiovisual context

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1416 Effect of Microfiltration on the Composition and Ripening of Iranian Fetta Cheese

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

Abstract:

The effect of Microfiltration (MF) on proteolysis, hardness, and flavor of Feta cheese during 6 mo of aging was determined. Raw skim milk was microfiltered two-fold in two cheese making trials. In trial 1, four vats of cheese were made in 1 d using unconcentrated milk (1X), 1.26X, 1.51X, and 1.82X Concentration Factors (CF). Casein-(CN)-to-fat ratio was constant among treatments. Proteolysis during cheese aging decreased with increasing CF due to either limitation of substrate availability for chymosin due to low moisture in the nonfat substance (MNFS), inhibition of chymosin activity by high molecular weight milk serum proteins, such as α2-macroglobulin, retained in the cheese or low residual chymosin in the cheese. Hardness of fresh cheese increased, and cheese flavor intensity decreased with increasing CF. In trial 2, the 1X and 1.8X CF were compared directly. Changes made in the cheese making procedure for the 1.8X CF (more chymosin and less cooking) increased the MNFS and made proteolysis during aging more comparable for the 1X and 1.8X cheeses. The significant difference in cheese hardness due to CF in trial 1 was eliminated in trial 2. In a triangle test, panelists could not differentiate between the 1X and 1.8X cheeses. Therefore, increasing chymosin and making the composition of the two cheeses more similar allowed production of aged Fetta cheese from milk concentrated up to 1.8X by MF that was not perceived as different from aged feta cheese produced without MF.

Keywords: feta cheese, microfiltration, concentration factor, proteolysis

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1415 The Impact of Efflux Pump Inhibitor on the Activity of Benzosiloxaboroles and Benzoxadiboroles against Gram-Negative Rods

Authors: Agnieszka E. Laudy, Karolina Stępien, Sergiusz Lulinski, Krzysztof Durka, Stefan Tyski

Abstract:

1,3-dihydro-1-hydroxy-2,1-benzoxaborole and its derivatives are a particularly interesting group of synthetic agents and were successfully employed in supramolecular chemistry medicine. The first important compounds, 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole and 5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole were identified as potent antifungal agents. In contrast, (S)-3-(aminomethyl)-7-(3-hydroxypropoxy)-1-hydroxy-1,3-dihydro-2,1-benzoxaborole hydrochloride is in the second phase of clinical trials as a drug for the treatment of Gram-negative bacterial infections of the Enterobacteriaceae family and Pseudomonas aeruginosa. Equally important and difficult task is to search for compounds active against Gram-negative bacilli, which have multi-drug-resistance efflux pumps actively removing many of the antibiotics from bacterial cells. We have examined whether halogen-substituted benzoxaborole-based derivatives and their analogues possess antibacterial activity and are substrates for multi-drug-resistance efflux pumps. The antibacterial activity of 1,3-dihydro-3-hydroxy-1,1-dimethyl-1,2,3-benzosiloxaborole and 10 halogen-substituted its derivatives, as well as 1,2-phenylenediboronic acid and 3 synthesised fluoro-substituted its analogs, were evaluated. The activity against the reference strains of Gram-positive (n=5) and Gram-negative bacteria (n=10) was screened by the disc-diffusion test (0.4 mg of tested compounds was applied onto paper disc). The minimal inhibitory concentration values and the minimal bactericidal concentration values were estimated according to The Clinical and Laboratory Standards Institute and The European Committee on Antimicrobial Susceptibility Testing recommendations. During the minimal inhibitory concentration values determination with or without phenylalanine-arginine beta-naphthylamide (50 mg/L) efflux pump inhibitor, the concentrations of tested compounds ranged 0.39-400 mg/L in the broth medium supplemented with 1 mM magnesium sulfate. Generally, the studied benzosiloxaboroles and benzoxadiboroles showed a higher activity against Gram-positive cocci than against Gram-negative rods. Moreover, benzosiloxaboroles have the higher activity than benzoxadiboroles compounds. In this study, we demonstrated that substitution (mono-, di- or tetra-) of 1,3-dihydro-3-hydroxy-1,1-dimethyl-1,2,3-benzosiloxaborole with halogen groups resulted in an increase in antimicrobial activity as compared to the parent substance. Interestingly, the 6,7-dichloro-substituted parent substance was found to be the most potent against Gram-positive cocci: Staphylococcus sp. (minimal inhibitory concentration 6.25 mg/L) and Enterococcus sp. (minimal inhibitory concentration 25 mg/L). On the other hand, mono- and dichloro-substituted compounds were the most actively removed by efflux pumps present in Gram-negative bacteria mainly from Enterobacteriaceae family. In the presence of efflux pump inhibitor the minimal inhibitory concentration values of chloro-substituted benzosiloxaboroles decreased from 400 mg/L to 3.12 mg/L. Of note, the highest increase in bacterial susceptibility to tested compounds in the presence of phenylalanine-arginine beta-naphthylamide was observed for 6-chloro-, 6,7-dichloro- and 6,7-difluoro-substituted benzosiloxaboroles. In the case of Escherichia coli, Enterobacter cloacae and P. aeruginosa strains at least a 32-fold decrease in the minimal inhibitory concentration values of these agents were observed. These data demonstrate structure-activity relationships of the tested derivatives and highlight the need for further search for benzoxaboroles and related compounds with significant antimicrobial properties. Moreover, the influence of phenylalanine-arginine beta-naphthylamide on the susceptibility of Gram-negative rods to studied benzosiloxaboroles indicate that some tested agents are substrates for efflux pumps in Gram-negative rods.

Keywords: antibacterial activity, benzosiloxaboroles, efflux pumps, phenylalanine-arginine beta-naphthylamide

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1414 Exploration of Abuse of Position for Sexual Gain by UK Police

Authors: Terri Cole, Fay Sweeting

Abstract:

Abuse of position for sexual gain by police is defined as behavior involving individuals taking advantage of their role to pursue a sexual or improper relationship. Previous research has considered whether it involves ‘bad apples’ - individuals with poor moral ethos or ‘bad barrels’ – broader organizational flaws which may unconsciously allow, minimize, or do not effectively deal with such behavior. Low level sexual misconduct (e.g., consensual sex on duty) is more common than more serious offences (e.g., rape), yet the impact of such behavior can have severe implications not only for those involved but can also negatively undermine public confidence in the police. This ongoing, collaborative research project has identified variables from 514 historic case files from 35 UK police forces in order to identify potential risk indicators which may lead to such behavior. Quantitative analysis using logistic regression and the Cox proportion hazard model has resulted in the identification of specific risk factors of significance in prediction. Factors relating to both perpetrator background such as a history of intimate partner violence, debt, and substance misuse coupled with in work behavior such as misusing police systems increase the risk. Findings are able to provide pragmatic recommendations for those tasked with identifying potential or investigating suspected perpetrators of misconduct.

Keywords: abuse of position, forensic psychology, misconduct, sexual abuse

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1413 Transformers in Gene Expression-Based Classification

Authors: Babak Forouraghi

Abstract:

A genetic circuit is a collection of interacting genes and proteins that enable individual cells to implement and perform vital biological functions such as cell division, growth, death, and signaling. In cell engineering, synthetic gene circuits are engineered networks of genes specifically designed to implement functionalities that are not evolved by nature. These engineered networks enable scientists to tackle complex problems such as engineering cells to produce therapeutics within the patient's body, altering T cells to target cancer-related antigens for treatment, improving antibody production using engineered cells, tissue engineering, and production of genetically modified plants and livestock. Construction of computational models to realize genetic circuits is an especially challenging task since it requires the discovery of flow of genetic information in complex biological systems. Building synthetic biological models is also a time-consuming process with relatively low prediction accuracy for highly complex genetic circuits. The primary goal of this study was to investigate the utility of a pre-trained bidirectional encoder transformer that can accurately predict gene expressions in genetic circuit designs. The main reason behind using transformers is their innate ability (attention mechanism) to take account of the semantic context present in long DNA chains that are heavily dependent on spatial representation of their constituent genes. Previous approaches to gene circuit design, such as CNN and RNN architectures, are unable to capture semantic dependencies in long contexts as required in most real-world applications of synthetic biology. For instance, RNN models (LSTM, GRU), although able to learn long-term dependencies, greatly suffer from vanishing gradient and low-efficiency problem when they sequentially process past states and compresses contextual information into a bottleneck with long input sequences. In other words, these architectures are not equipped with the necessary attention mechanisms to follow a long chain of genes with thousands of tokens. To address the above-mentioned limitations of previous approaches, a transformer model was built in this work as a variation to the existing DNA Bidirectional Encoder Representations from Transformers (DNABERT) model. It is shown that the proposed transformer is capable of capturing contextual information from long input sequences with attention mechanism. In a previous work on genetic circuit design, the traditional approaches to classification and regression, such as Random Forrest, Support Vector Machine, and Artificial Neural Networks, were able to achieve reasonably high R2 accuracy levels of 0.95 to 0.97. However, the transformer model utilized in this work with its attention-based mechanism, was able to achieve a perfect accuracy level of 100%. Further, it is demonstrated that the efficiency of the transformer-based gene expression classifier is not dependent on presence of large amounts of training examples, which may be difficult to compile in many real-world gene circuit designs.

Keywords: transformers, generative ai, gene expression design, classification

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1412 New Challenge: Reduction of Aflatoxin M1 Residues in Cow’s Milk by MilBond Dietary Hydrated Sodium Calcium Aluminosilicate (HSCAS) and Its Effect on Milk Composition

Authors: A. Aly Salwa, H. Diekmann, S. Hafiz Ragaa, DG Abo Elhassan

Abstract:

This study was aimed to evaluate the effect of Milbond (HSCAS) on aflatoxin M1 in artificially contaminated cows milk. Chemisorption compounds used in this experiment were MIlBond, hydrated sodium calcium aluminosilicate (HSCAS). Raw cow milk were artificially exposed to aflatoxin M1 in a concentration of 100 ppb) with addition of Nilbond at 0.5, 1, 2 and 3 % at room temperature for 30 minutes. Aflatoxin M1 was decreased more than 95% by HSCAS at 2%. Milk composition consist of protein, fat, lactose, solid non fat and total solid were affected by addition of some adsorbents were not significantly affected (p 0.05). Tthis method did not involve degrading the toxin, milk may be free from toxin degradation products and is safe for consumption. In addition, the added material may be easily separated from milk after the substance adsorbs the toxin. Thus, this method should be developed by further researches for determining effects of these compounds on functional properties of milk. The ability of hydrated sodium calcium aluminosilicate to prevent or reduce the level of aflatoxin MI residues in milk is critically needed. This finding has important implications, because milk is ultimately consumed by humans and animals, and the reduction of aflatoxin contamination in the milk could have an important impact on their health.

Keywords: aflatoxin M1, Hydrated sodium calcium aluminium silicate, detoxification, raw cow milk

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1411 Comparative Analysis of Feature Extraction and Classification Techniques

Authors: R. L. Ujjwal, Abhishek Jain

Abstract:

In the field of computer vision, most facial variations such as identity, expression, emotions and gender have been extensively studied. Automatic age estimation has been rarely explored. With age progression of a human, the features of the face changes. This paper is providing a new comparable study of different type of algorithm to feature extraction [Hybrid features using HAAR cascade & HOG features] & classification [KNN & SVM] training dataset. By using these algorithms we are trying to find out one of the best classification algorithms. Same thing we have done on the feature selection part, we extract the feature by using HAAR cascade and HOG. This work will be done in context of age group classification model.

Keywords: computer vision, age group, face detection

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1410 Therapeutic Effect of Indane 1,3-Dione Derivatives in the Restoration of Insulin Resistance in Human Liver Cells and in Db/Db Mice Model: Biochemical, Physiological and Molecular Insights of Investigation

Authors: Gulnaz Khan, Meha F. Aftab, Munazza Murtaza, Rizwana S. Waraich

Abstract:

Advanced glycation end products (AGEs) precursor and its abnormal accumulation cause damage to various tissues and organs. AGEs have pathogenic implication in several diseases including diabetes. Existing AGEs inhibitors are not in clinical use, and there is a need for development of novel inhibitors. The present investigation aimed at identifying the novel AGEs inhibitors and assessing their mechanism of action for treating insulin resistance in mice model of diabetes. Novel derivatives of benzylidene of indan-1,3-dione were synthesized. The compounds were selected to study their action mechanism in improving insulin resistance, in vitro, in human hepatocytes and murine adipocytes and then, in vivo, in mice genetic model of diabetes (db/db). Mice were treated with novel derivatives of benzylidene of indane 1,3-dione. AGEs mediated ROS production was measured by dihydroethidium fluorescence assay. AGEs level in the serum of treated mice was observed by ELISA. Gene expression of receptor for AGEs (RAGE), PPAR-gamma, TNF-alpha and GLUT-4 was evaluated by RT-PCR. Glucose uptake was measured by fluorescent method. Microscopy was used to analyze glycogen synthesis in muscle. Among several derivatives of benzylidene of indan-1,3-dione, IDD-24, demonstrated highest inhibition of AGESs. IDD-24 significantly reduced AGEs formation and expression of receptor for advanced glycation end products (RAGE) in fat, liver of db/db mice. Suppression of AGEs mediated ROS production was also observed in hepatocytes and fat cell, after treatment with IDD-24. Glycogen synthesis was increased in muscle tissue of mice treated with IDD-24. In adipocytes, IDD-24 prevented AGEs induced reduced glucose uptake. Mice treated with IDD-24 exhibited increased glucose tolerance, serum adiponectin levels and decreased insulin resistance. The result of present study suggested that IDD-24 can be a possible treatment target to address glycotoxins induced insulin resistance.

Keywords: advance glycation end product, hyperglycemia, indan-1, 3-dione, insulin resistance

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1409 The Influence of Alvar Aalto on the Early Work of Álvaro Siza

Authors: Eduardo Jorge Cabral dos Santos Fernandes

Abstract:

The expression ‘Porto School’, usually associated with an educational institution, the School of Fine Arts of Porto, is applied for the first time with the sense of an architectural trend by Nuno Portas in a text published in 1983. The expression is used to characterize a set of works by Porto architects, in which common elements are found, namely the desire to reuse languages and forms of the German and Dutch rationalism of the twenties, using the work of Alvar Aalto as a mediation for the reinterpretation of these models. In the same year, Álvaro Siza classifies the Finnish architect as a miscegenation agent who transforms experienced models and introduces them to different realities in a text published in Jornal de Letras, Artes e Ideias. The influence of foreign models and their adaptation to the context has been a recurrent theme in Portuguese architecture, which finds important contributions in the writings of Alexandre Alves Costa, at this time. However, the identification of these characteristics in Siza’s work is not limited to the Portuguese theoretical production: it is the recognition of this attitude towards the context that leads Kenneth Frampton to include Siza in the restricted group of architects who embody Critical Regionalism (in his book Modern architecture: a critical history). For Frampton, his work focuses on the territory and on the consequences of the intervention in the context, viewing architecture as a tectonic fact rather than a series of scenographic episodes and emphasizing site-specific aspects (topography, light, climate). Therefore, the motto of this paper is the dichotomous opposition between foreign influences and adaptation to the context in the early work of Álvaro Siza (designed in the sixties) in which the influence (theoretical, methodological, and formal) of Alvar Aalto manifests itself in the form and the language: the pool at Quinta da Conceição, the Seaside Pools and the Tea House (three works in Leça da Palmeira) and the Lordelo Cooperative (in Porto). This work is part of a more comprehensive project, which considers several case studies throughout the Portuguese architect's vast career, built in Portugal and abroad, in order to obtain a holistic view.

Keywords: Alvar Aalto, Álvaro Siza, foreign influences, adaptation to the context

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1408 The Effect and Mechanisms of Electroacupuncture on Motion Sickness in Mice

Authors: Chanya Inprasit, Yi-Wen Lin

Abstract:

Motion sickness (MS) is an acute disorder that occurs in healthy persons without considering gender, age or ethnicity worldwide. All signs and symptoms of MS are the results of confliction and mismatch among neural signal inputs. It is known that no singular remedy works for everybody, and electroacupuncture (EA) is one of the popular alternative therapies used for MS. Our study utilized a mouse model in order to exclude any psychological factors of MS and EA. Mice lack an emetic reflex. Therefore pica behavior, which is a normal consumption of non-nutritive substances, was found to measure the response of MS in mice. In the laboratory, Kaolin was used as a non-nutrient food substance instead of natural substances lacking nutritional value such as wood, cloth, charcoal, soil or grass. It was hypothesized that EA treatment could reduce the symptoms of MS through the TRPV1 pathways. The results of pica behavior showed a significantly increased intake of kaolin in the MS group throughout the experiment period. Moreover, the Kaolin intake of the EA group decreased to the average baseline of the control group. There was no recorded difference in the food and water intake of each group. The results indicated an increase of the TRPV1, pERK, pJNK and pmTOR protein levels in the thalamus after MS stimulation, and a significant decrease in the EA group compared with that of the control group. These findings suggest that TRPV1 pathways are associated in MS mechanisms and can be reduced by EA.

Keywords: electroacupuncture, motion sickness, Thalamus, TRPV1

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1407 A Web-Based Systems Immunology Toolkit Allowing the Visualization and Comparative Analysis of Publically Available Collective Data to Decipher Immune Regulation in Early Life

Authors: Mahbuba Rahman, Sabri Boughorbel, Scott Presnell, Charlie Quinn, Darawan Rinchai, Damien Chaussabel, Nico Marr

Abstract:

Collections of large-scale datasets made available in public repositories can be used to identify and fill gaps in biomedical knowledge. But first, these data need to be made readily accessible to researchers for analysis and interpretation. Here a collection of transcriptome datasets was made available to investigate the functional programming of human hematopoietic cells in early life. Thirty two datasets were retrieved from the NCBI Gene Expression Omnibus (GEO) and loaded in a custom, interactive web application called the Gene Expression browser (GXB), designed for visualization and query of integrated large-scale data. Multiple sample groupings and gene rank lists were created based on the study design and variables in each dataset. Web links to customized graphical views can be generated by users and subsequently be used to graphically present data in manuscripts for publication. The GXB tool also enables browsing of a single gene across datasets, which can provide information on the role of a given molecule across biological systems. The dataset collection is available online. As a proof-of-principle, one of the datasets (GSE25087) was re-analyzed to identify genes that are differentially expressed by regulatory T cells in early life. Re-analysis of this dataset and a cross-study comparison using multiple other datasets in the above mentioned collection revealed that PMCH, a gene encoding a precursor of melanin-concentrating hormone (MCH), a cyclic neuropeptide, is highly expressed in a variety of other hematopoietic cell types, including neonatal erythroid cells as well as plasmacytoid dendritic cells upon viral infection. Our findings suggest an as yet unrecognized role of MCH in immune regulation, thereby highlighting the unique potential of the curated dataset collection and systems biology approach to generate new hypotheses which can be tested in future mechanistic studies.

Keywords: early-life, GEO datasets, PMCH, interactive query, systems biology

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1406 Stochastic Repair and Replacement with a Single Repair Channel

Authors: Mohammed A. Hajeeh

Abstract:

This paper examines the behavior of a system, which upon failure is either replaced with certain probability p or imperfectly repaired with probability q. The system is analyzed using Kolmogorov's forward equations method; the analytical expression for the steady state availability is derived as an indicator of the system’s performance. It is found that the analysis becomes more complex as the number of imperfect repairs increases. It is also observed that the availability increases as the number of states and replacement probability increases. Using such an approach in more complex configurations and in dynamic systems is cumbersome; therefore, it is advisable to resort to simulation or heuristics. In this paper, an example is provided for demonstration.

Keywords: repairable models, imperfect, availability, exponential distribution

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1405 Statistical Pattern Recognition for Biotechnological Process Characterization Based on High Resolution Mass Spectrometry

Authors: S. Fröhlich, M. Herold, M. Allmer

Abstract:

Early stage quantitative analysis of host cell protein (HCP) variations is challenging yet necessary for comprehensive bioprocess development. High resolution mass spectrometry (HRMS) provides a high-end technology for accurate identification alongside with quantitative information. Hereby we describe a flexible HRMS assay platform to quantify HCPs relevant in microbial expression systems such as E. Coli in both up and downstream development by means of MVDA tools. Cell pellets were lysed and proteins extracted, purified samples not further treated before applying the SMART tryptic digest kit. Peptides separation was optimized using an RP-UHPLC separation platform. HRMS-MSMS analysis was conducted on an Orbitrap Velos Elite applying CID. Quantification was performed label-free taking into account ionization properties and physicochemical peptide similarities. Results were analyzed using SIEVE 2.0 (Thermo Fisher Scientific) and SIMCA (Umetrics AG). The developed HRMS platform was applied to an E. Coli expression set with varying productivity and the corresponding downstream process. Selected HCPs were successfully quantified within the fmol range. Analysing HCP networks based on pattern analysis facilitated low level quantification and enhanced validity. This approach is of high relevance for high-throughput screening experiments during upstream development, e.g. for titer determination, dynamic HCP network analysis or product characterization. Considering the downstream purification process, physicochemical clustering of identified HCPs is of relevance to adjust buffer conditions accordingly. However, the technology provides an innovative approach for label-free MS based quantification relying on statistical pattern analysis and comparison. Absolute quantification based on physicochemical properties and peptide similarity score provides a technological approach without the need of sophisticated sample preparation strategies and is therefore proven to be straightforward, sensitive and highly reproducible in terms of product characterization.

Keywords: process analytical technology, mass spectrometry, process characterization, MVDA, pattern recognition

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1404 Hamilton-Jacobi Treatment of Damped Motion

Authors: Khaled I. Nawafleh

Abstract:

In this work, we apply the method of Hamilton-Jacobi to obtain solutions of Hamiltonian systems in classical mechanics with two certain structures: the first structure plays a central role in the theory of time-dependent Hamiltonians, whilst the second is used to treat classical Hamiltonians, including dissipation terms. It is proved that the generalization of problems from the calculus of variation methods in the nonstationary case can be obtained naturally in Hamilton-Jacobi formalism. Then, another expression of geometry of the Hamilton Jacobi equation is retrieved for Hamiltonians with time-dependent and frictional terms. Both approaches shall be applied to many physical examples.

Keywords: Hamilton-Jacobi, time dependent lagrangians, dissipative systems, variational principle

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