Search results for: genes expression
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2479

Search results for: genes expression

199 Geothermal Resources to Ensure Energy Security During Climate Change

Authors: Debasmita Misra, Arthur Nash

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Energy security and sufficiency enables the economic development and welfare of a nation or a society. Currently, the global energy system is dominated by fossil fuels, which is a non-renewable energy resource, which renders vulnerability to energy security. Hence, many nations have begun augmenting their energy system with renewable energy resources, such as solar, wind, biomass and hydro. However, with climate change, how sustainable are some of the renewable energy resources in the future is a matter of concern. Geothermal energy resources have been underexplored or underexploited in global renewable energy production and security, although it is gaining attractiveness as a renewable energy resource. The question is, whether geothermal energy resources are more sustainable than other renewable energy resources. High-temperature reservoirs (> 220 °F) can produce electricity from flash/dry steam plants as well as binary cycle production facilities. Most of the world’s high enthalpy geothermal resources are within the seismo-tectonic belt. However, exploration for geothermal energy is of great importance in conventional geothermal systems in order to improve its economic viability. In recent years, there has been an increase in the use and development of several exploration methods for geo-thermal resources, such as seismic or electromagnetic methods. The thermal infrared band of the Landsat can reflect land surface temperature difference, so the ETM+ data with specific grey stretch enhancement has been used to explore underground heat water. Another way of exploring for potential power is utilizing fairway play analysis for sites without surface expression and in rift zones. Utilizing this type of analysis can improve the success rate of project development by reducing exploration costs. Identifying the basin distribution of geologic factors that control the geothermal environment would help in identifying the control of resource concentration aside from the heat flow, thus improving the probability of success. The first step is compiling existing geophysical data. This leads to constructing conceptual models of potential geothermal concentrations which can then be utilized in creating a geodatabase to analyze risk maps. Geospatial analysis and other GIS tools can be used in such efforts to produce spatial distribution maps. The goal of this paper is to discuss how climate change may impact renewable energy resources and how could a synthesized analysis be developed for geothermal resources to ensure sustainable and cost effective exploitation of the resource.

Keywords: exploration, geothermal, renewable energy, sustainable

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198 Characterization of a Lipolytic Enzyme of Pseudomonas nitroreducens Isolated from Mealworm's Gut

Authors: Jung-En Kuan, Whei-Fen Wu

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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

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197 Covid-19 Lockdown Experience of Elderly Female as Reflected in Their Artwork

Authors: Liat Shamri-Zeevi, Neta Ram-Vlasov

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Today the world as a whole is attempting to cope with the COVID-19, which has affected all facets of personal and social life from country-wide confinement to maintaining social distance and taking protective measures to maintain hygiene. One of the populations faced with the most severe restrictions is seniors. Various studies have shown that creativity plays a crucial role in dealing with crisis events. Painting - regardless of media - allows for emotional and cognitive processing of these situations, and enables the expression of experiences in a tangible creative way that conveys and endows meaning to the artwork. The current study was conducted in Israel immediately after a 6-week lockdown. It was designed to specifically examine the impact of the COVID-19 pandemic on the quality of life of elderly women as reflected in their artworks. The sample was composed of 21 Israeli women aged 60-90, in good mental health (without diagnosed dementia or Alzheimer's), all of whom were Hebrew-speaking, and retired with an extended family, who indicated that they painted and had engaged in artwork on an ongoing basis throughout the lockdown (from March 12 to May 30, 2020). The participants' artworks were collected, and a semi-structured in-depth interview was conducted that lasted one to two hours. The participants were asked about their feelings during the pandemic and the artworks they produced during this time, and completed a questionnaire on well-being and mental health. The initial analysis of the interviews and artworks revealed themes related to the specific role of each piece of artwork. The first theme included notions that the artwork was an activity and a framework for doing, which supported positive emotions, and provided a sense of vitality during the closure. Most of the participants painted images of nature and growth which were ascribed concrete and symbolic meaning. The second theme was that the artwork enabled the processing of difficult and /or conflicting emotions related to the situation, including anxiety about death and loneliness that were symbolically expressed in the artworks, such as images of the Corona virus and the respiratory machines. The third theme suggested that the time and space prompted by the lockdown gave the participants time for a gathering together of the self, and freed up time for creative activities. Many participants stated that they painted more and more frequently during the Corona lockdown. At the conference, additional themes and findings will be presented.

Keywords: Corona virus, artwork, quality of life of elderly

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196 Luteolin Exhibits Anti-Diabetic Effects by Increasing Oxidative Capacity and Regulating Anti-Oxidant Metabolism

Authors: Eun-Young Kwon, Myung-Sook Choi, Su-Jung Cho, Ji-Young Choi, So Young Kim, Youngji Han

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Overweight and obesity have been linked to a low-grade chronic inflammatory response and an increased risk of developing metabolic syndrome including insulin resistance, type 2 diabetes mellitus and certain types of cancers. Luteolin is a dietary flavonoid with anti-inflammatory, anti-oxidant, anti-cancer and anti-diabetic properties. However, little is known about the detailed mechanism associated with the effect of luteolin on inflammation-related obesity and its complications. The aim of the present study was to reveal the anti-diabetic effect of luteolin in diet-induced obesity mice using “transcriptomics” tool. Thirty-nine male C57BL/6J mice (4-week-old) were randomly divided into 3 groups and were fed normal diet, high-fat diet (HFD, 20% fat) and HFD+0.005% (w/w) luteolin for 16 weeks. Luteolin improved insulin resistance, as measured by HOMA-IR and glucose tolerance, along with preservation action of pancreatic β-cells, compared to the HFD group. Luteoiln was significantly decreased the levels of leptin and ghrelin that play a pivotal role in energy balance, and the macrophage low-grade inflammation marker sCD163 (soluble Cd antigen 163) in plasma. Activities of hepatic anti-oxidant enzymes (catalase and glutathione peroxidase) were increased, while the levels of plasma transaminase (GOT and GPT) and oxidative damage markers (hepatic mitochondria H2O2 and TBARS) were markedly decreased by luteolin supplementation. In addition, luteolin increased oxidative capacity and fatty acid utilization by presenting decrease in enzyme activities of citrate synthase, cytochrome C oxidase and β-hydroxyacyl CoA dehydrogenase and UCP3 gene expression compared to high-fat diet. Moreover, our microarray results of muscle also revealed down-regulated gene expressions associated with TCA cycle by HFD were reversed to normal level by luteolin treatment. Taken together, our results indicate that luteolin is one of bioactive components for improving insulin resistance by increasing oxidative capacity, modulating anti-oxidant metabolism and suppressing inflammatory signaling cascades in diet-induced obese mice. These results provide possible therapeutic targets for prevention and treatment of diet-induced obesity and its complications.

Keywords: anti-oxidant metabolism, diabetes, luteolin, oxidative capacity

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195 iPSCs More Effectively Differentiate into Neurons on PLA Scaffolds with High Adhesive Properties for Primary Neuronal Cells

Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.

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Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.

Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid

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194 Isolation of Bacterial Species with Potential Capacity for Siloxane Removal in Biogas Upgrading

Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

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Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

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193 An Empty Canvas is Full

Authors: Joonha Park

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This essay examines the Soviet Artist Pavel Korin’s artistic pursuit towards his life-long project, “Requiem/Passing of the Rus,” framing the funeral of Tikhon, the last great defender of the Russian orthodox Church during the Great purge, as the final moment of “Rus,” which is the identity of the Russian people that built up in the 1000 year of history behind Russian Orthodoxy. Korin’s project remains in the form of a series of 29 man-sized portraits and a monumental blank canvas. Born in a family dedicated to iconography, Korin witnessed the historic drama during Stalin’s terror; therefore, he tried to convey the nation’s mourning for the disappearance of “Rus” and disapproval of the Soviet notion of atheism. Yet, due to Korin’s success as a state artist, many believed that the political pressure led Korin to give up his belief and controversy arose over the fact that Korin left the canvas blank. The empty 40-square-meter canvas, which remains untouched in his studio since 1930, supports this theory to an extent. However, resources such as Korin’s notes, primary accounts from his fellow Soviet Artists, and testimonies from his wife suggested that this assumption is incorrect. Moreover, Korin’s uninterrupted relationship with the church and the religious attributes in his commissioned works were brought up as evidence of Korin’s continued belief. The empty canvas not only represents Korin’s discontentment towards the repression and the hardships that the Orthodox Church experienced, but also depicts the identity that coexisted with the Church in order to bequeath this idea to future generations. The faultless canvas surrounded by the striking 29 portraits is a symbol of the highest spirit, similar to that of the iconography paintings placed in every Russian house that unites the Russian people till this day, therefore one can deduce that the legacy of “Requiem” is still relevant to the Russian people even under freedom of religious expression. Consequently, “Requiem” was on display at the Tretyakov Gallery for the first time in public in 2013 even though Korin started creating this piece in 1925, extolling Korin not only as an artist but also as a historian; by recording the turmoil of the Great Oppression, Korin exhibited the social responsibility universal to artists across time and space. In this essay, the legacy Korin left behind, both to his contemporaries and his posterity is reevaluated through the lens of his works, unfinished as they were.

Keywords: Pavel Korin, Art History, Art, Russia, Soviet Union, Requiem, Russian orthodox church, Treytyakov gallery, contemporary art, socialist realism, Maxim Gorky

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192 Intercultural Competence among Jewish and Arab Students Studying Together in an Academic Institution in Israel

Authors: Orly Redlich

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Since the establishment of the state of Israel, and as a result of various events that led to it, Jewish citizens and Arab citizens of the state have been in constant conflict, which finds its expression in most levels of life. Therefore, the attitude of one group member to the other group members is mostly tense, loaded, and saturated with mutual suspicion. Within this reality, in many higher education institutions in Israel, Jews and Arabs meet with each other intensively and for several years. For some students, this is their first opportunity for a meaningful cross-cultural encounter. These intercultural encounters, which allow positive interactions between members of different cultural groups, may contribute to the formation of "intercultural competence" which means long-term change in knowledge, attitudes, and behavior towards 'the other culture'. The current study examined the concept of the ‘other’ among Jewish and Arab students studying together and their "intercultural competence". The study also examined whether there is a difference in the perception of the ‘other’ between students studying in different academic programs, and between students taking academic courses on multiculturalism. This quantitative study was conducted among 274 Arab and Jewish students studying together, for bachelors or master's degree, in various academic programs at the Israel Academic College of Ramat-Gan. The background data of the participants are varied, in terms of religion, origin, religiosity, employment status, living area, and marital status. The main hypothesis is that academic, social, and intercultural encounters between Jewish and Arab students, who attend college together, will be a significant factor in building "intercultural competence". Additionally, the existence of "intercultural competence" has been linked to demographic characteristics of the students, as well as the nature of intercultural encounters between Jews and Arabs in a higher education institution. The dependent variables were measured by a self-report questionnaire, using the components of '"intercultural competence"' among students, which are: 1. Cognitive knowledge of the ‘others’, 2. Feelings towards the ‘others’, 3. Change in attitudes towards the 'others', and 4. Change in behavior towards the ‘others’. The findings indicate a higher "intercultural competence" among Arab students than Jews; it was also found higher level of "intercultural competence" among Educational Counseling students than the other respondents. The importance of this research lies in finding the means to develop "intercultural competence" among Jewish and Arab students, which may reduce prejudice and stereotypes towards the other culture and may even prevent occurrences of alienation and violence in cross-cultural encounters in Israel.

Keywords: cross-cultural learning, intercultural competence, Jewish and Arab students, multiculturalism

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191 Neuro-Epigenetic Changes on Diabetes Induced-Synaptic Fidelity in Brain

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

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

Keywords: epigenetics, cognition, chaperones, DNA methylation

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190 Anti-Arthritic Effect of a Herbal Diet Formula Comprising Fruits of Rosa Multiflora and Flowers of Lonicera Japonica

Authors: Brian Chi Yan Cheng, Hui Guo, Tao Su, Xiu‐qiong Fu, Ting Li, Zhi‐ling Yu

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Rheumatoid arthritis (RA) affects around 1% of the globe population. Yet, there is still no cure for RA. Toll-like receptor 4 (TLR4) signalling has been found to be involved in the pathogenesis of RA, making it a potential therapeutic target for RA treatment. A herbal formula (RL) consisting of fruits of Rosa Multiflora (Eijitsu rose) and flowers of Lonicera Japonica (Japanese honeysuckle) has been used in treating various inflammatory disorders for more than a thousand year. Both of them are rich sources of nutrients and bioactive phytochemicals, which can be used in producing different food products and supplements. In this study, we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis (CIA) in rats and investigate the involvement of TLR4 signaling in the mode of action of RL. Anti-arthritic efficacy was evaluated using CIA rats induced by bovine type II collagen. The treatment groups were treated with RL (82.5, 165, and 330 mg/kg bw per day, p.o.) or positive control indomethacin (0.25 mg/kg bw per day, p.o.) for 35 days. Clinical signs (hind paw volume and arthritis severity scores), changes in serum inflammatory mediators, pro-/antioxidant status, histological and radiographic changes of joints were investigated. Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats. The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats, murine RAW264.7 macrophages and human THP-1 monocytic cells. The severity of arthritis in the CIA rats was significantly attenuated by RL. Antioxidant status, histological score and radiographic score were efficiently improved by RL. RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats. RL significantly inhibited the production of various pro-inflammatory mediators, the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines. RL possesses anti-arthritic effect on collagen-induced arthritis in rats. The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum. The inhibition of the TAK1/NF-κB and TAK1/MAPK pathways participate in the anti-arthritic effects of RL. This provides a pharmacological justification for the dietary use of RL in the control of various arthritic diseases. Further investigation should be done to develop RL into a anti-arthritic food products and/or supplements.

Keywords: japanese honeysuckle, rheumatoid arthritis, rosa multiflora, rosehip

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189 Assessment of the Effect of Ethanolic Leaf Extract of Annona squamosa L. on Den Induced Hepatocellular Carcinoma in Experimental Animals

Authors: Vanitha Varadharaj, Vijalakshmi Krishnamurthy

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Annona squamosa Linn, commonly known as Sugar apple, belonging to the family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The alkaloid and flavonoids present in Annona squamosa leaf has proved to have antioxidant activity. The present work has been planned to investigate the effect of ethanolic leaf extract of Annona squamosa leaf on Den Induced wistar albino rats. The study was carried out to analyze the biochemical Parmeters like Total Proteins, Bilirubin, Enzymatic and Non –Enzymatic enzymes, Marker enzymes and Tumor markers in serum and also the histopathological studies in liver is carried out in control and DEN induced rats. Supplementation of ELAS (Ethanolic Leaf Extract Of Annona squamosa) reduced the liver weight and also reduced the tumour incidence. Chemoprevention group showed near normal values of bilirubin when compared with the control rats. Total protein was decreased in the cancer bearing group and on treatment with the extract the levels of protein were restored. Both in pre and post treatment group, the activities of enzymatic antioxidants such as superoxide dismutase, catalase, and Glutathione peroxidase were increased but in pre treated animals it was more effective than post treated animals. The non- enzymatic antioxidants such as vitamin C and vitamin E were brought back to normal level significantly in post and pre treated animals. Activities of marker enzymes such as SGOT, SGPT, ALP, γ GT were significantly elevated in the serum of cancer animals and the values returned to normal after treatment with the extract suggesting the hepato protective effect of the extract. Lipid peroxide was found to be elevated in the cancer induced group. This condition was brought back to the normal in the pre and post treated animals with ELAS. Histological examination also confirmed the anti- carcinogenic potential of ELAS, Cancer induced groups had a triple fold increase in their AFP values when compared to other groups. DEN treatment increased the level of AFP expression while ELAS partially counteracted the effect of it. So the scientific validation obtained from this study may pave way to many budding scientists to find new drugs from Annona squamosa for various ailments.

Keywords: annona squamosa, biochemical parmeters, cancer, leaf extract

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188 "If It Bleeds It Leads” the Visual Witnessing Trauma Phenomenon among Journalists: An Analysis of Various Media Images from East Africa

Authors: Lydia Ouma Radoli

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The paradox of documenting history through visuals that objectify gruesome images to depict the prominence of stories intrigues media researchers. In East Africa, the topic has been captured in a variety of media frames, but scantly in scholarly work. This paper adopts Visual Rhetoric and Framing Theories to tease out the drivers behind the criteria for the selection of violent visuals. The paper projects that quantitative and qualitative literature regarding journalists’ personal and work-related exposure to PSTD will give insights into the concept of trauma journalism - reporting of horrific events, e.g., violent crime and terror. The data will be collected through methods such as document analysis (photographs and videos) and in-depth interviews to summarize the informational contents with respect to the research objectives and questions. The study is hinged on the background that the criterion for news production is constructed from the idea that ‘if there’s violence, conflict, and death involved, the story gets top priority.’ The anticipated outcome is to establish trauma experiences of visual rhetors, suggest mitigations, and address gaps in academic research. The findings of the study will sustain the critical role of visual rhetors. Further, media practitioners may find the study useful in assessing the effects and values of visual witnessing. Historically, the criterion for visual news production has been that if there’s violence, conflict, and death involved, the story gets top priority. To capture the goriness of the images, media theorists and sociologists have used the expression: “If it bleeds, it leads.” The statement assumes that audiences are attracted to pictures that show violent images. Further, research on visual aspects of Television news has shown its ability to hold viewers’ attention and cause aggression. This paper samples images and narratives from Journalists who have covered trauma-related events. The samples are indicative of the problem under study, which depicts journalists exposed to traumatic events as not receiving any Psycho-social support within newsrooms. It is hoped that the study could inform policy and practice within developing countries through the interpretations of theoretical and empirical explanations of existing trauma phenomena among journalists.

Keywords: visual-witnessing, media culture, visual rhetoric, imaging violence in East Africa

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187 Targeting Methionine Metabolism In Gastric Cancer; Promising To Improve Chemosensetivity With Non-hetrogeneity

Authors: Nigatu Tadesse, Li Juan, Liuhong Ming

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Gastric cancer (GC) is the fifth most common and fourth deadly cancer in the world with limited treatment options at late advanced stage in which surgical therapy is not recommended with chemotherapy remain as the mainstay of treatment. However, the occurrence of chemoresistance as well as intera-tumoral and inter-tumoral heterogeneity of response to targeted and immunotherapy underlined a clear unmet treatment need in gastroenterology. Several molecular and cellular alterations ascribed for chemo resistance in GC including cancer stem cells (CSC) and tumor microenvironment (TME) remodeling. Cancer cells including CSC bears higher metabolic demand and major changes in TME involves alterations of gut microbiota interacting with nutrients metabolism. Metabolic upregulation in lipids, carbohydrates, amino acids, fatty acids biosynthesis pathways identified as a common hall mark in GC. Metabolic addiction to methionine metabolism occurs in many cancer cells to promote the biosynthesis of S-Adenosylmethionine (SAM), a universal methyl donor molecule for high rate of transmethylation in GC and promote cell proliferation. Targeting methionine metabolism found to promotes chemo-sensitivity with treatment non-heterogeneity. Methionine restriction (MR) promoted the arrest of cell cycle at S/G2 phase and enhanced downregulation of GC cells resistance to apoptosis (including ferroptosis), which suggests the potential of synergy with chemotherapies acting at S-phase of the cell cycle as well as inducing cell apoptosis. Accumulated evidences showed both the biogenesis as well as intracellular metabolism of exogenous methionine could be safe and effective target for therapy either alone or in combination with chemotherapies. This review article provides an over view of the upregulation in methionine biosynthesis pathway and the molecular signaling through the PI3K/Akt/mTOR-c-MYC axis to promote metabolic reprograming through activating the expression of L-type aminoacid-1 (LAT1) transporter and overexpression of Methionine adenosyltransferase 2A(MAT2A) for intercellular metabolic conversion of exogenous methionine to SAM in GC, and the potential of targeting with novel therapeutic agents such as methioninase (METase), Methionine adenosyltransferase 2A (MAT2A), c-MYC, methyl like transferase 16 (METTL16) inhibitors that are currently under clinical trial development stages and future perspectives.

Keywords: gastric cancer, methionine metabolism, pi3k/akt/mtorc1-c-myc axis, gut microbiota, MAT2A, c-MYC, METTL16, methioninase

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186 Biosynthesis of a Nanoparticle-Antibody Phthalocyanine Photosensitizer for Use in Targeted Photodynamic Therapy of Cervical Cancer

Authors: Elvin P. Chizenga, Heidi Abrahamse

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Cancer cell resistance to therapy is the main cause of treatment failures and the poor prognosis of cancer convalescence. The progression of cervical cancer to other parts of the genitourinary system and the reported recurrence rates are overwhelming. Current treatments, including surgery, chemo and radiation have been inefficient in eradicating the tumor cells. These treatments are also associated with poor prognosis and reduced quality of life, including fertility loss. This has inspired the need for the development of new treatment modalities to eradicate cervical cancer successfully. Photodynamic Therapy (PDT) is a modern treatment modality that induces cell death by photochemical interactions of light and a photosensitizer, which in the presence of molecular oxygen, yields a set of chemical reactions that generate Reactive Oxygen Species (ROS) and other free radical species causing cell damage. Enhancing PDT using modified drug delivery can increase the concentration of the photosensitizer in the tumor cells, and this has the potential to maximize its therapeutic efficacy. In cervical cancer, all infected cells constitutively express genes of the E6 and E7 HPV viral oncoproteins, resulting in high concentrations of E6 and E7 in the cytoplasm. This provides an opportunity for active targeting of cervical cancer cells using immune-mediated drug delivery to maximize therapeutic efficacy. The use of nanoparticles in PDT has also proven effective in enhancing therapeutic efficacy. Gold nanoparticles (AuNps) in particular, are explored for their use in biomedicine due to their biocompatibility, low toxicity, and enhancement of drug uptake by tumor cells. In this present study, a biomolecule comprising of AuNPs, anti-E6 monoclonal antibodies, and Aluminium Phthalocyanine photosensitizer was synthesized for use in targeted PDT of cervical cancer. The AuNp-Anti-E6-Sulfonated Aluminium Phthalocyanine mix (AlPcSmix) photosensitizing biomolecule was synthesized by coupling AuNps and anti-E6 monoclonal antibodies to the AlPcSmix via Polyethylene Glycol (PEG) chemical links. The final product was characterized using Transmission Electron Microscope (TEM), Zeta Potential, Uv-Vis Spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), and X-ray diffraction (XRD), to confirm its chemical structure and functionality. To observe its therapeutic role in treating cervical cancer, cervical cancer cells, HeLa cells were seeded in 3.4 cm² diameter culture dishes at a concentration of 5x10⁵ cells/ml, in vitro. The cells were treated with varying concentrations of the photosensitizing biomolecule and irradiated using a 673.2 nm wavelength of laser light. Post irradiation cellular responses were performed to observe changes in morphology, viability, proliferation, cytotoxicity, and cell death pathways induced. Dose-Dependent response of the cells to treatment was demonstrated as significant morphologic changes, increased cytotoxicity, and decreased cell viability and proliferation This study presented a synthetic biomolecule for targeted PDT of cervical cancer. The study suggested that PDT using this AuNp- Anti-E6- AlPcSmix photosensitizing biomolecule is a very effective treatment method for the eradication of cervical cancer cells, in vitro. Further studies in vivo need to be conducted to support the use of this biomolecule in treating cervical cancer in clinical settings.

Keywords: anti-E6 monoclonal antibody, cervical cancer, gold nanoparticles, photodynamic therapy

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185 Tasting Terroir: A Gourmet Adventure in Food and Wine Tourism

Authors: Sunita Boro, Saurabh Kumar Dixit

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Terroir, an intricate fusion of geography, climate, soil, and human expertise, has long been acknowledged as a defining factor in the character of wines and foods. This research embarks on an exploration of terroir's profound influence on gastronomic tourism, shedding light on the intricate interplay between the physical environment and culinary artistry. Delving into the intricate science of terroir, we scrutinize its role in shaping the sensory profiles of wines and foods, emphasizing the profound impact of specific regions on flavor, aroma, and texture. We deploy a multifaceted methodology, amalgamating sensory analysis, chemical profiling, geographical information systems, and qualitative interviews to unearth the nuanced nuances of terroir expression. Through an exhaustive review of the literature, we elucidate the historical roots of terroir, unveil the intricate cultural dimensions shaping it, and provide a comprehensive examination of prior studies in the field. Our findings underscore the pivotal role of terroir in promoting regional identities, enhancing the economic viability of locales, and attracting gastronomic tourists. The paper also dissects the marketing strategies employed to promote terroir-driven food and wine experiences. We elucidate the utilization of storytelling, branding, and collaborative endeavors in fostering a robust terroir-based tourism industry. This elucidates both the potential for innovation and the challenges posed by oversimplification or misrepresentation of terroir. Our research spotlights the intersection of terroir and sustainability, emphasizing the significance of environmentally conscious practices in terroir-driven productions. We discern the harmonious relationship between sustainable agriculture, terroir preservation, and responsible tourism, encapsulating the essence of ecological integrity in gastronomic tourism. Incorporating compelling case studies of regions and businesses excelling in the terroir-based tourism realm, we offer in-depth insights into successful models and strategies, with an emphasis on their replicability and adaptability to various contexts. Ultimately, this paper not only contributes to the scholarly understanding of terroir's role in the world of food and wine tourism but also provides actionable recommendations for stakeholders to leverage terroir's allure, preserve its authenticity, and foster sustainable and enriching culinary tourism experiences.

Keywords: terroir, food tourism, wine tourism, sustainability

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184 Cultural and Natural Heritage Conservation by GIS Tourism Inventory System Project

Authors: Gamze Safak, Umut Arslanoglu

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Cultural and tourism conservation and development zones and tourism centers are the boundaries declared for the purpose of protecting, using, and evaluating the sectoral development and planned development in areas where historical and cultural values are heavily involved and/or where tourism potential is high. The most rapidly changing regions in Turkey are tourism areas, especially the coastal areas. Planning these regions is not about only an economic gain but also a natural and physical environment and refers to a complex process. If the tourism sector is not well controlled, excessive use of natural resources and wrong location choices may cause damage to natural areas, historical values, and socio-cultural structure. Since the strategic decisions taken in the environmental order and zoning plans, which are the means of guiding the physical environment of the Ministry of Culture and Tourism, which have the authority to make plans in tourism centers, are transformed into plan decisions that find the spatial expression, comprehensive evaluation of all kinds of data, following the historical development and based on the correct and current data is required. In addition, the authority has a number of competences in tourism promotion as well as the authority to plan, leading to the necessity of taking part in the applications requiring complex analysis such as the management and integration of the country's economic, political, social and cultural resources. For this purpose, Tourism Inventory System (TES) project, which consists of a series of subsystems, has been developed in order to solve complex planning and method problems in the management of site-related information. The scope of the project is based on the integration of numerical and verbal data in the regions within the jurisdiction of the authority, and the monitoring of the historical development of urban planning studies, making the spatial data of the institution easily accessible, shared, questionable and traceable in international standards. A dynamic and continuous system design has been put into practice by utilizing the advantage of the use of Geographical Information Systems in the planning process to play a role in making the right decisions, revealing the tools of social, economic, cultural development, and preservation of natural and cultural values. This paper, which is prepared by the project team members in TES (Tourism Inventory System), will present a study regarding the applicability of GIS in cultural and natural heritage conservation.

Keywords: cultural conservation, GIS, geographic information system, tourism inventory system, urban planning

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183 The Impact of Hosting an On-Site Vocal Concert in Preschool on Music Inspiration and Learning Among Preschoolers

Authors: Meiying Liao, Poya Huang

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The aesthetic domain is one of the six major domains in the Taiwanese preschool curriculum, encompassing visual arts, music, and dramatic play. Its primary objective is to cultivate children’s abilities in exploration and awareness, expression and creation, and response and appreciation. The purpose of this study was to explore the effects of hosting a vocal music concert on aesthetic inspiration and learning among preschoolers in a preschool setting. The primary research method employed was a case study focusing on a private preschool in Northern Taiwan that organized a school-wide event featuring two vocalists. The concert repertoires included children’s songs, folk songs, and arias performed in Mandarin, Hakka, English, German, and Italian. In addition to professional performances, preschool teachers actively participated by presenting a children’s song. A total of 5 classes, comprising approximately 150 preschoolers, along with 16 teachers and staff, participated in the event. Data collection methods included observation, interviews, and documents. Results indicated that both teachers and children thoroughly enjoyed the concert, with high levels of acceptance when the program was appropriately designed and hosted. Teachers reported that post-concert discussions with children revealed the latter’s ability to recall people, events, and elements observed during the performance, expressing their impressions of the most memorable segments. The concert effectively achieved the goals of the aesthetic domain, particularly in fostering response and appreciation. It also inspired preschoolers’ interest in music. Many teachers noted an increased desire for performance among preschoolers after exposure to the concert, with children imitating the performers and their expressions. Remarkably, one class extended this experience by incorporating it into the curriculum, autonomously organizing a high-quality concert in the music learning center. Parents also reported that preschoolers enthusiastically shared their concert experiences at home. In conclusion, despite being a single event, the positive responses from preschoolers towards the music performance suggest a meaningful impact. These experiences extended into the curriculum, as firsthand exposure to performances allowed teachers to deepen related topics, fostering a habit of autonomous learning in the designated learning centers.

Keywords: concert, early childhood music education, aesthetic education, music develpment

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182 Exploring the Link between Hoarding Disorder and Trauma: A Scoping Review

Authors: Murray Anderson, Galina Freed, Karli Jahn

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Trauma is increasingly recognized as an important construct that has health implications for those who struggle with various mental health issues. For those individuals who meet the criteria for a diagnosis of hoarding disorder (HD), many have experienced some form of trauma. Further, some of the therapeutic interventions for those with HD can further perpetuate or magnify the experience of trauma. Therefore, the aim of this scoping review is to identify and document the nature and extent of research evidence related to trauma as it connects with HD. This review was guided by the questions, ‘How can our understanding of the trauma cycle help us to better appreciate the experiences of individuals who hoard, and how will a trauma informed lens inform the interventions for hoarding disorder? A comprehensive literature search was performed to identify original studies that contained the words “hoarding” and “trauma.” PsychINFO”,''EBSCO host,” “CINAHL” and “PubMed” were searched between January 2005 and April 2021. Articles were screened by three reviewers. Data extracted included publication date, demographics, study design, type of analysis, and noted connections between hoarding and trauma. Of the 329 articles, all duplicates, articles on hoardings of animals, articles not in English, and those without full-text availability were removed. Five categories were found in the remaining 45 articles, including (a) traumatic and stressful life events; (b) the link between posttraumatic stress disorder, trauma, and hoarding; (c) the relationships between different comorbidities, trauma, and hoarding; (d) the lack of early emotional expression and other forms of parental deprivation; and (e) the role of attachment. Lastly, the literature explains how the links between hoarding and trauma are difficult to study due to the highly stigmatized identities with this population. The review provided strong support for the connections between the experience of trauma and HD. What is missing from the literature is the use of a trauma-informed lens to better account for the ways in which hoarding disorder is understood. Other missing pieces in the literature are the potential uses of a trauma-informed lens to enhance the therapeutic process, to understand and reduce treatment attrition, and to improve treatment outcomes. The application of a trauma informed lens could improve our understanding of effective interactions among clients, families, and communities and improve the education around hoarding-related matters. Exploring the connections between trauma and HD can improve therapeutic delivery and destigmatize the experience of dealing with clutter and hoarding concerns. This awareness can also provide health care professionals with both the language and skills to liberate them from a reductionist view on HD.

Keywords: hoarding, attachment, parental deprivation, trauma

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181 Angiomotin Regulates Integrin Beta 1-Mediated Endothelial Cell Migration and Angiogenesis

Authors: Yuanyuan Zhang, Yujuan Zheng, Giuseppina Barutello, Sumako Kameishi, Kungchun Chiu, Katharina Hennig, Martial Balland, Federica Cavallo, Lars Holmgren

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Angiogenesis describes that new blood vessels migrate from pre-existing ones to form 3D lumenized structure and remodeling. During directional migration toward the gradient of pro-angiogenic factors, the endothelial cells, especially the tip cells need filopodia to sense the environment and exert the pulling force. Of particular interest are the integrin proteins, which play an essential role in focal adhesion in the connection between migrating cells and extracellular matrix (ECM). Understanding how these biomechanical complexes orchestrate intrinsic and extrinsic forces is important for our understanding of the underlying mechanisms driving angiogenesis. We have previously identified Angiomotin (Amot), a member of Amot scaffold protein family, as a promoter for endothelial cell migration in vitro and zebrafish models. Hence, we established inducible endothelial-specific Amot knock-out mice to study normal retinal angiogenesis as well as tumor angiogenesis. We found that the migration ratio of the blood vessel network to the edge was significantly decreased in Amotec- retinas at postnatal day 6 (P6). While almost all the Amot defect tip cells lost migration advantages at P7. In consistence with the dramatic morphology defect of tip cells, there was a non-autonomous defect in astrocytes, as well as the disorganized fibronectin expression pattern correspondingly in migration front. Furthermore, the growth of transplanted LLC tumor was inhibited in Amot knockout mice due to fewer vasculature involved. By using MMTV-PyMT transgenic mouse model, there was a significantly longer period before tumors arised when Amot was specifically knocked out in blood vessels. In vitro evidence showed that Amot binded to beta-actin, Integrin beta 1 (ITGB1), Fibronectin, FAK, Vinculin, major focal adhesion molecules, and ITGB1 and stress fibers were distinctly induced by Amot transfection. Via traction force microscopy, the total energy (force indicater) was found significantly decreased in Amot knockdown cells. Taken together, we propose that Amot is a novel partner of the ITGB1/Fibronectin protein complex at focal adhesion and required for exerting force transition between endothelial cell and extracellular matrix.

Keywords: angiogenesis, angiomotin, endothelial cell migration, focal adhesion, integrin beta 1

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180 Understanding Different Facets of Chromosome Abnormalities: A 17-year Cytogenetic Study and Indian Perspectives

Authors: Lakshmi Rao Kandukuri, Mamata Deenadayal, Suma Prasad, Bipin Sethi, Srinadh Buragadda, Lalji Singh

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Worldwide; at least 7.6 million children are born annually with severe genetic or congenital malformations and among them 90% of these are born in mid and low-income countries. Precise prevalence data are difficult to collect, especially in developing countries, owing to the great diversity of conditions and also because many cases remain undiagnosed. The genetic and congenital disorder is the second most common cause of infant and childhood mortality and occurs with a prevalence of 25-60 per 1000 births. The higher prevalence of genetic diseases in a particular community may, however, be due to some social or cultural factors. Such factors include the tradition of consanguineous marriage, which results in a higher rate of autosomal recessive conditions including congenital malformations, stillbirths, or mental retardation. Genetic diseases can vary in severity, from being fatal before birth to requiring continuous management; their onset covers all life stages from infancy to old age. Those presenting at birth are particularly burdensome and may cause early death or life-long chronic morbidity. Genetic testing for several genetic diseases identifies changes in chromosomes, genes, or proteins. The results of a genetic test can confirm or rule out a suspected genetic condition or help determine a person's chance of developing or passing on a genetic disorder. Several hundred genetic tests are currently in use and more are being developed. Chromosomal abnormalities are the major cause of human suffering, which are implicated in mental retardation, congenital malformations, dysmorphic features, primary and secondary amenorrhea, reproductive wastage, infertility neoplastic diseases. Cytogenetic evaluation of patients is helpful in the counselling and management of affected individuals and families. We present here especially chromosomal abnormalities which form a major part of genetic disease burden in India. Different programmes on chromosome research and human reproductive genetics primarily relate to infertility since this is a major public health problem in our country, affecting 10-15 percent of couples. Prenatal diagnosis of chromosomal abnormalities in high-risk pregnancies helps in detecting chromosomally abnormal foetuses. Such couples are counselled regarding the continuation of pregnancy. In addition to the basic research, the team is providing chromosome diagnostic services that include conventional and advanced techniques for identifying various genetic defects. Other than routine chromosome diagnosis for infertility, also include patients with short stature, hypogonadism, undescended testis, microcephaly, delayed developmental milestones, familial, and isolated mental retardation, and cerebral palsy. Thus, chromosome diagnostics has found its applicability not only in disease prevention and management but also in guiding the clinicians in certain aspects of treatment. It would be appropriate to affirm that chromosomes are the images of life and they unequivocally mirror the states of human health. The importance of genetic counseling is increasing with the advancement in the field of genetics. The genetic counseling can help families to cope with emotional, psychological, and medical consequences of genetic diseases.

Keywords: India, chromosome abnormalities, genetic disorders, cytogenetic study

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179 Constructivist Grounded Theory of Intercultural Learning

Authors: Vaida Jurgile

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Intercultural learning is one of the approaches taken to understand the cultural diversity of the modern world and to accept changes in cultural identity and otherness and the expression of tolerance. During intercultural learning, students develop their abilities to interact and communicate with their group members. These abilities help to understand social and cultural differences, to form one’s identity, and to give meaning to intercultural learning. Intercultural education recognizes that a true understanding of differences and similarities of another culture is necessary in order to lay the foundations for working together with others, which contributes to the promotion of intercultural dialogue, appreciation of diversity, and cultural exchange. Therefore, it is important to examine the concept of intercultural learning, revealed through students’ learning experiences and understanding of how this learning takes place and what significance this phenomenon has in higher education. At a scientific level, intercultural learning should be explored in order to uncover the influence of cultural identity, i.e., intercultural learning should be seen in a local context. This experience would provide an opportunity to learn from various everyday intercultural learning situations. Intercultural learning can be not only a form of learning but also a tool for building understanding between people of different cultures. The research object of the study is the process of intercultural learning. The aim of the dissertation is to develop a grounded theory of the process of learning in an intercultural study environment, revealing students’ learning experiences. The research strategy chosen in this study is a constructivist grounded theory (GT). GT is an inductive method that seeks to form a theory by applying the systematic collection, synthesis, analysis, and conceptualization of data. The targeted data collection was based on the analysis of data provided by previous research participants, which revealed the need for further research participants. During the research, only students with at least half a year of study experience, i.e., who have completed at least one semester of intercultural studies, were purposefully selected for the research. To select students, snowballing sampling was used. 18 interviews were conducted with students representing 3 different fields of sciences (social sciences, humanities, and technology sciences). In the process of intercultural learning, language expresses and embodies cultural reality and a person’s cultural identity. It is through language that individual experiences are expressed, and the world in which Others exist is perceived. The increased emphasis is placed on the fact that language conveys certain “signs’ of communication and perception with cultural value, enabling the students to identify the Self and the Other. Language becomes an important tool in the process of intercultural communication because it is only through language that learners can communicate, exchange information, and understand each other. Thus, in the process of intercultural learning, language either promotes interpersonal relationships with foreign students or leads to mutual rejection.

Keywords: intercultural learning, grounded theory, students, other

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178 Structural and Binding Studies of Peptidyl-tRNA Hydrolase from Pseudomonas aeruginosa Provide a Platform for the Structure Based Inhibitor Design against Peptidyl-tRNA Hydrolase

Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

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177 Re-interpreting Ruskin with Respect to the Wall

Authors: Anjali Sadanand, R. V. Nagarajan

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Architecture morphs with advances in technology and the roof, wall, and floor as basic elements of a building, follow in redefining themselves over time. Their contribution is bound by time and held by design principles that deal with function, sturdiness, and beauty. Architecture engages with people to give joy through its form, material, design structure, and spatial qualities. This paper attempts to re-interpret John Ruskin’s “Seven lamps of Architecture” in the context of the architecture of the modern and present period. The paper focuses on the “wall” as an element of study in this context. Four of Ruskin’s seven lamps will be discussed, namely beauty, truth, life, and memory, through examples of architecture ranging from modernism to contemporary architecture of today. The study will focus on the relevance of Ruskin’s principles to the “wall” in specific, in buildings of different materials and over a range of typologies from all parts of the world. Two examples will be analyzed for each lamp. It will be shown that in each case, there is relevance to the significance of Ruskin’s lamps in modern and contemporary architecture. Nature to which Ruskin alludes to for his lamp of “beauty” is found in the different expressions of interpretation used by Corbusier in his Villa Stein façade based on proportion found in nature and in the direct expression of Toyo Ito in his translation of an understanding of the structure of trees into his façade design of the showroom for a Japanese bag boutique. “Truth” is shown in Mies van der Rohe’s Crown Hall building with its clarity of material and structure and Studio Mumbai’s Palmyra House, which celebrates the use of natural materials and local craftsmanship. “Life” is reviewed with a sustainable house in Kerala by Ashrams Ravi and Alvar Aalto’s summer house, which illustrate walls as repositories of intellectual thought and craft. “Memory” is discussed with Charles Correa’s Jawahar Kala Kendra and Venturi’s Vana Venturi house and discloses facades as text in the context of its materiality and iconography. Beauty is reviewed in Villa Stein and Toyo Ito’s Branded Retail building in Tokyo. The paper thus concludes that Ruskin’s Lamps can be interpreted in today’s context and add richness to meaning to the understanding of architecture.

Keywords: beauty, design, facade, modernism

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176 Identification of Potent and Selective SIRT7 Anti-Cancer Inhibitor via Structure-Based Virtual Screening and Molecular Dynamics Simulation

Authors: Md. Fazlul Karim, Ashik Sharfaraz, Aysha Ferdoushi

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Background: Computational medicinal chemistry approaches are used for designing and identifying new drug-like molecules, predicting properties and pharmacological activities, and optimizing lead compounds in drug development. SIRT7, a nicotinamide adenine dinucleotide (NAD+)-dependent deacylase which regulates aging, is an emerging target for cancer therapy with mounting evidence that SIRT7 downregulation plays important roles in reversing cancer phenotypes and suppressing tumor growth. Activation or altered expression of SIRT7 is associated with the progression and invasion of various cancers, including liver, breast, gastric, prostate, and non-small cell lung cancer. Objectives: The goal of this work was to identify potent and selective bioactive candidate inhibitors of SIRT7 by in silico screening of small molecule compounds obtained from Nigella sativa (N. sativa). Methods: SIRT7 structure was retrieved from The Research Collaboratory for Structural Bioinformatics Protein Data Bank (RCSB PDB), and its active site was identified using CASTp and metaPocket. Molecular docking simulation was performed with PyRx 0.8 virtual screening software. Drug-likeness properties were tested using SwissADME and pkCSM. In silico toxicity was evaluated by Osiris Property Explorer. Bioactivity was predicted by Molinspiration software. Antitumor activity was screened for Prediction of Activity Spectra for Substances (PASS) using Way2Drug web server. Molecular dynamics (MD) simulation was carried out by Desmond v3.6 package. Results: A total of 159 bioactive compounds from the N. Sativa were screened against the SIRT7 enzyme. Five bioactive compounds: chrysin (CID:5281607), pinocembrin (CID:68071), nigellidine (CID:136828302), nigellicine (CID:11402337), and epicatechin (CID:72276) were identified as potent SIRT7 anti-cancer candidates after docking score evaluation and applying Lipinski's Rule of Five. Finally, MD simulation identified Chrysin as the top SIRT7 anti-cancer candidate molecule. Conclusion: Chrysin, which shows a potential inhibitory effect against SIRT7, can act as a possible anti-cancer drug candidate. This inhibitor warrants further evaluation to check its pharmacokinetics and pharmacodynamics properties both in vitro and in vivo.

Keywords: SIRT7, antitumor, molecular docking, molecular dynamics simulation

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175 Glucose Uptake Rate of Insulin-Resistant Human Liver Carcinoma Cells (IR/HepG2) by Flavonoids from Enicostema littorale via IR/IRS1/AKT Pathway

Authors: Priyanka Mokashi, Aparna Khanna, Nancy Pandita

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Diabetes mellitus is a chronic metabolic disorder which will be the 7th leading cause of death by 2030. The current line of treatment for the diabetes mellitus is oral antidiabetic drugs (biguanides, sulfonylureas, meglitinides, thiazolidinediones and alpha-glycosidase inhibitors) and insulin therapy depending upon the type 1 or type 2 diabetes mellitus. But, these treatments have their disadvantages, ranging from the developing of resistance to the drugs and adverse effects caused by them. Alternative to these synthetic agents, natural products provides a new insight for the development of more efficient and safe drugs due to their therapeutic values. Enicostema littorale blume (A. Raynal) is a traditional Indian plant belongs to the Gentianaceae family. It is widely distributed in Asia, Africa, and South America. There are few reports on Swrtiamarin, major component of this plant for its antidiabetic activity. However, the antidiabetic activity of flavonoids from E. littorale and their mechanism of action have not yet been elucidated. Flavonoids have a positive relationship with disease prevention and can act on various molecular targets and regulate different signaling pathways in pancreatic β-cells, adipocytes, hepatocytes and skeletal myofibers. They may exert beneficial effects in diabetes by (i) improving hyperglycemia through regulation of glucose metabolism in hepatocytes; (ii) enhancing insulin secretion and reducing apoptosis and promoting proliferation of pancreatic β-cells; (iii) increasing glucose uptake in hepatocytes, skeletal muscle and white adipose tissue (iv) reducing insulin resistance, inflammation and oxidative stress. Therefore, we have isolated four flavonoid rich fractions, Fraction A (FA), Fraction B (FB), Fraction C (FC), Fraction D (FD) from crude alcoholic hot (AH) extract from E. littorale, identified by LC/MS. Total eight flavonoids were identified on the basis of fragmentation pattern. Flavonoid FA showed the presence of swertisin, isovitexin, and saponarin; FB showed genkwanin, quercetin, isovitexin, FC showed apigenin, swertisin, quercetin, 5-O-glucosylswertisin and 5-O-glucosylisoswertisin whereas FD showed the presence of swertisin. Further, these fractions were assessed for their antidiabetic activity on stimulating glucose uptake in insulin-resistant HepG2 cell line model (IR/HepG2). The results showed that FD containing C-glycoside Swertisin has significantly increased the glucose uptake rate of IR/HepG2 cells at the concentration of 10 µg/ml as compared to positive control Metformin (0.5mM) which was determined by glucose oxidase- peroxidase method. It has been reported that enhancement of glucose uptake of cells occurs due the translocation of Glut4 vesicles to cell membrane through IR/IRS1/AKT pathway. Therefore, we have studied expressions of three genes IRS1, AKT and Glut4 by real-time PCR to evaluate whether they follow the same pathway or not. It was seen that the glucose uptake rate has increased in FD treated IR/HepG2 cells due to the activation of insulin receptor substrate-1 (IRS1) followed by protein kinase B (AKT) through phosphoinositide 3-kinase (PI3K) leading to translocation of Glut 4 vesicles to cell membrane, thereby enhancing glucose uptake and insulin sensitivity of insulin resistant HepG2 cells. Hence, the up-regulation indicated the mechanism of action through which FD (Swertisin) acts as antidiabetic candidate in the treatment of type 2 diabetes mellitus.

Keywords: E. littorale, glucose transporter, glucose uptake rate, insulin resistance

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174 Informational Habits and Ideology as Predictors for Political Efficacy: A Survey Study of the Brazilian Political Context

Authors: Pedro Cardoso Alves, Ana Lucia Galinkin, José Carlos Ribeiro

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Political participation, can be a somewhat tricky subject to define, not in small part due to the constant changes in the concept fruit of the effort to include new forms of participatory behavior that go beyond traditional institutional channels. With the advent of the internet and mobile technologies, defining political participation has become an even more complicated endeavor, given de amplitude of politicized behaviors that are expressed throughout these mediums, be it in the very organization of social movements, in the propagation of politicized texts, videos and images, or in the micropolitical behaviors that are expressed in daily interaction. In fact, the very frontiers that delimit physical and digital spaces have become ever more diluted due to technological advancements, leading to a hybrid existence that is simultaneously physical and digital, not limited, as it once was, to the temporal limitations of classic communications. Moving away from those institutionalized actions of traditional political behavior, an idea of constant and fluid participation, which occurs in our daily lives through conversations, posts, tweets and other digital forms of expression, is discussed. This discussion focuses on the factors that precede more direct forms of political participation, interpreting the relation between informational habits, ideology, and political efficacy. Though some of the informational habits can be considered political participation, by some authors, a distinction is made to establish a logical flow of behaviors leading to participation, that is, one must gather and process information before acting on it. To reach this objective, a quantitative survey is currently being applied in Brazilian social media, evaluating feelings of political efficacy, social and economic issue-based ideological stances and informational habits pertaining to collection, fact-checking, and diversity of sources and ideological positions present in the participant’s political information network. The measure being used for informational habits relies strongly on a mix of information literacy and political sophistication concepts, bringing a more up-to-date understanding of information and knowledge production and processing in contemporary hybrid (physical-digital) environments. Though data is still being collected, preliminary analysis point towards a strong correlation between information habits and political efficacy, while ideology shows a weaker influence over efficacy. Moreover, social ideology and economic ideology seem to have a strong correlation in the sample, such intermingling between social and economic ideals is generally considered a red flag for political polarization.

Keywords: political efficacy, ideology, information literacy, cyberpolitics

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173 Establishment of Farmed Fish Welfare Biomarkers Using an Omics Approach

Authors: Pedro M. Rodrigues, Claudia Raposo, Denise Schrama, Marco Cerqueira

Abstract:

Farmed fish welfare is a very recent concept, widely discussed among the scientific community. Consumers’ interest regarding farmed animal welfare standards has significantly increased in the last years posing a huge challenge to producers in order to maintain an equilibrium between good welfare principles and productivity, while simultaneously achieve public acceptance. The major bottleneck of standard aquaculture is to impair considerably fish welfare throughout the production cycle and with this, the quality of fish protein. Welfare assessment in farmed fish is undertaken through the evaluation of fish stress responses. Primary and secondary stress responses include release of cortisol and glucose and lactate to the blood stream, respectively, which are currently the most commonly used indicators of stress exposure. However, the reliability of these indicators is highly dubious, due to a high variability of fish responses to an acute stress and the adaptation of the animal to a repetitive chronic stress. Our objective is to use comparative proteomics to identify and validate a fingerprint of proteins that can present an more reliable alternative to the already established welfare indicators. In this way, the culture conditions will improve and there will be a higher perception of mechanisms and metabolic pathway involved in the produced organism’s welfare. Due to its high economical importance in Portuguese aquaculture Gilthead seabream will be the elected species for this study. Protein extracts from Gilthead Seabream fish muscle, liver and plasma, reared for a 3 month period under optimized culture conditions (control) and induced stress conditions (Handling, high densities, and Hipoxia) are collected and used to identify a putative fish welfare protein markers fingerprint using a proteomics approach. Three tanks per condition and 3 biological replicates per tank are used for each analisys. Briefly, proteins from target tissue/fluid are extracted using standard established protocols. Protein extracts are then separated using 2D-DIGE (Difference gel electrophoresis). Proteins differentially expressed between control and induced stress conditions will be identified by mass spectrometry (LC-Ms/Ms) using NCBInr (taxonomic level - Actinopterygii) databank and Mascot search engine. The statistical analysis is performed using the R software environment, having used a one-tailed Mann-Whitney U-test (p < 0.05) to assess which proteins were differentially expressed in a statistically significant way. Validation of these proteins will be done by comparison of the RT-qPCR (Quantitative reverse transcription polymerase chain reaction) expressed genes pattern with the proteomic profile. Cortisol, glucose, and lactate are also measured in order to confirm or refute the reliability of these indicators. The identified liver proteins under handling and high densities induced stress conditions are responsible and involved in several metabolic pathways like primary metabolism (i.e. glycolysis, gluconeogenesis), ammonia metabolism, cytoskeleton proteins, signalizing proteins, lipid transport. Validition of these proteins as well as identical analysis in muscle and plasma are underway. Proteomics is a promising high-throughput technique that can be successfully applied to identify putative welfare protein biomarkers in farmed fish.

Keywords: aquaculture, fish welfare, proteomics, welfare biomarkers

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172 Cord Blood Hematopoietic Stem Cell Expansion Ability of Mesenchymal Stem Cells Isolated From Different Sources

Authors: Ana M. Lara, Manuela Llano, Felipe Gaitán, Rosa H. Bustos, Ana Maria Perdomo-Arciniegas, Ximena Bonilla

Abstract:

Umbilical cord blood is used as a source of progenitor and stem cells for the regeneration of the hematopoietic and immune system to treat patients with different hematological or non-hematological diseases. This stem cell source represents an advantage over the use of bone marrow or mobilized peripheral blood because it has a lower incidence rate of graft-versus-host disease, probably due to fewer immunological compatibility restrictions. However, its low cellular dose limits its use in pediatric patients. This work proposes the standardization of a cell expansion technique to compensate for the dose of infused cells through the ex-vivo manipulation of hematopoietic progenitor cells from umbilical cord blood before transplantation. The expansion model is carried out through co-cultures with mesenchymal stem cells (MSC) from bone marrow (BM) and less explored fetal tissues such as Wharton's jelly (WJ) and umbilical cord blood (UCB). Initially, a master cell bank of primary mesenchymal stem cells isolated from different sources was established and characterized following International Society of Cell Therapies (ISCT) indications. Additionally, we assessed the effect of a short 25 Gy cycle of gamma irradiation on cell cycle arrest of mesenchymal cells over the support capacity for the expansion of hematopoietic stem cells from umbilical cord blood was evaluated. The results show that co-cultures with MSC from WJ and UCB allow the cellular dose of HSPC to be maximized between 5 and 16 times having a similar support capacity as BM. In addition, was evaluated the hematopoietic stem progenitor cell's HSPC functionality through the evaluation of migration capacity, their differentiation capacity during culture time by flow cytometry to evaluate the expression of membrane markers associated with lineage-committed progenitors, their clonogenic potential, and the evaluation of secretome profile in the expansion process was evaluated. So far, the treatment with gamma irradiation maintains the hematopoietic support capacity of mesenchymal stem cells from the three sources studied compared to treatments without irradiation, favoring the use of fetal tissues that are generally waste to obtain mesenchymal cell lines for ex-vivo expansion systems. With the results obtained, a standardized protocol that will contribute to the development of ex-vivo expansion with MSC on a larger scale will be achieved, enabling its clinical use and expanding its application in adults.

Keywords: ex-vivo expansion, hematopoietic stem cells, hematopoietic stem cell transplantation, mesenchymal stem cells, umbilical cord blood

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171 Evaluation of a Driver Training Intervention for People on the Autism Spectrum: A Multi-Site Randomized Control Trial

Authors: P. Vindin, R. Cordier, N. J. Wilson, H. Lee

Abstract:

Engagement in community-based activities such as education, employment, and social relationships can improve the quality of life for individuals with Autism Spectrum Disorder (ASD). Community mobility is vital to attaining independence for individuals with ASD. Learning to drive and gaining a driver’s license is a critical link to community mobility; however, for individuals with ASD acquiring safe driving skills can be a challenging process. Issues related to anxiety, executive function, and social communication may affect driving behaviours. Driving training and education aimed at addressing barriers faced by learner drivers with ASD can help them improve their driving performance. A multi-site randomized controlled trial (RCT) was conducted to evaluate the effectiveness of an autism-specific driving training intervention for improving the on-road driving performance of learner drivers with ASD. The intervention was delivered via a training manual and interactive website consisting of five modules covering varying driving environments starting with a focus on off-road preparations and progressing through basic to complex driving skill mastery. Seventy-two learner drivers with ASD aged 16 to 35 were randomized using a blinded group allocation procedure into either the intervention or control group. The intervention group received 10 driving lessons with the instructors trained in the use of an autism-specific driving training protocol, whereas the control group received 10 driving lessons as usual. Learner drivers completed a pre- and post-observation drive using a standardized driving route to measure driving performance using the Driving Performance Checklist (DPC). They also completed anxiety, executive function, and social responsiveness measures. The findings showed that there were significant improvements in driving performance for both the intervention (d = 1.02) and the control group (d = 1.15). However, the differences were not significant between groups (p = 0.614) or study sites (p = 0.842). None of the potential moderator variables (anxiety, cognition, social responsiveness, and driving instructor experience) influenced driving performance. This study is an important step toward improving community mobility for individuals with ASD showing that an autism-specific driving training intervention can improve the driving performance of leaner drivers with ASD. It also highlighted the complexity of conducting a multi-site design even when sites were matched according to geography and traffic conditions. Driving instructors also need more and clearer information on how to communicate with learner drivers with restricted verbal expression.

Keywords: autism spectrum disorder, community mobility, driving training, transportation

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170 Application of Thermoplastic Microbioreactor to the Single Cell Study of Budding Yeast to Decipher the Effect of 5-Hydroxymethylfurfural on Growth

Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

Abstract:

Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast

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