Search results for: gene diversity

Authors: T. Sadunishvili, L. Kutateladze, T. Urushadze, R. Khvedelidze, N. Zakariashvili, M. Jobava, G. Kvesitadze

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Multiple repeated soil-climatic zones in Georgia determines the diversity of microorganisms. Hundreds of microscopic fungi of different genera have been isolated from different ecological niches, including some extreme environments. Biosynthetic ability of microscopic fungi has been studied. Trichoderma ressei, representative of the Ascomycetes secrete cellulolytic and xylanolytic enzymes that act in synergy to hydrolyze polysaccharide polymers to glucose, xylose and arabinose, which can be fermented to biofuels. The other mesophilic strains producing cellulases are Allesheria terrestris, Chaetomium thermophile, Fusarium oxysporium, Piptoporus betulinus, Penicillium echinulatum, P. purpurogenum, Aspergillus niger, A. wentii, A. versicolor, A. fumigatus etc. In the majority of the cases the cellulases produced by strains of genus Aspergillus usually have high β-glucosidase activity and average endoglucanases levels (with some exceptions), whereas strains representing Trichoderma have high endo enzyme and low β-glucosidase, and hence has limited efficiency in cellulose hydrolysis. Six producers of stable cellulases and xylanases from mesophilic and thermophilic fungi have been selected. By optimization of submerged cultivation conditions, high activities of cellulases and xylanases were obtained. For enzymes purification, their sedimentation by organic solvents such as ethyl alcohol, acetone, isopropanol and by ammonium sulphate in different ratios have been carried out. Best results were obtained with precipitation by ethyl alcohol (1:3.5) and ammonium sulphate. The yields of enzyme according to cellulase activities were 80-85% in both cases. Cellulase activity of enzyme preparation obtained from the strain Trichoderma viride X 33 is 126 U/g, from the strain Penicillium canescence D 85–185U/g and from the strain Sporotrichum pulverulentum T 5-0 110 U/g. Cellulase activity of enzyme preparation obtained from the strain Aspergillus sp. Av10 is 120 U/g, xylanase activity of enzyme preparation obtained from the strain Aspergillus niger A 7-5–1155U/g and from the strain Aspergillus niger Aj 38-1250 U/g. Optimum pH and temperature of operation and thermostability, of the enzyme preparations, were established. The efficiency of hydrolyses of different agricultural residues by the microscopic fungi cellulases has been studied. The glucose yield from the residues as a result of enzymatic hydrolysis is highly determined by the ratio of enzyme to substrate, pH, temperature, and duration of the process. Hydrolysis efficiency was significantly increased as a result of different pretreatment of the residues by different methods. Acknowledgement: The Study was supported by the ISTC project G-2117, funded by Korea.

Keywords: cellulase, xylanase, microscopic fungi, enzymatic hydrolysis

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Authors: Sahar El Shair, Laura Greco, William Reay, Murray Cairns

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Background: Rheumatoid arthritis (RA) is a chronic, systematic, inflammatory, autoimmune disease that involves damages to joints and erosions to the associated bones and cartilage, resulting in reduced physical function and disability. RA is a multifactorial disorder influenced by heterogenous genetic and environmental factors. Whilst different medications have proven successful in reducing inflammation associated with RA, they often come with significant side effects and limited efficacy. To address this, the novel pharmagenic enrichment score (PES) algorithm was tested in self-reported RA patients from the UK Biobank (UKBB), which is a cohort of predominantly European ancestry, and identified individuals with a high genetic risk in clinically actionable biological pathways to identify novel opportunities for precision interventions and drug repurposing to treat RA. Methods and materials: Genetic association data for rheumatoid arthritis was derived from publicly available genome-wide association studies (GWAS) summary statistics (N=97173). The PES framework exploits competitive gene set enrichment to identify pathways that are associated with RA to explore novel treatment opportunities. This data is then integrated into WebGestalt, Drug Interaction database (DGIdb) and DrugBank databases to identify existing compounds with existing use or potential for repurposed use. The PES for each of these candidates was then profiled in individuals with RA in the UKBB (Ncases = 3,719, Ncontrols = 333,160). Results A total of 209 pathways with known drug targets after multiple testing correction were identified. Several pathways, including interferon gamma signaling and TID pathway (which relates to a chaperone that modulates interferon signaling), were significantly associated with self-reported RA in the UKBB when adjusting for age, sex, assessment centre month and location, RA polygenic risk and 10 principal components. These pathways have a major role in RA pathogenesis, including autoimmune attacks against certain citrullinated proteins, synovial inflammation, and bone loss. Encouragingly, many also relate to the mechanism of action of existing RA medications. The analyses also revealed statistically significant association between RA polygenic scores and self-reported RA with individual PES scorings, highlighting the potential utility of the PES algorithm in uncovering additional genetic insights that could aid in the identification of individuals at risk for RA and provide opportunities for more targeted interventions. Conclusions In this study, pharmacologically annotated genetic risk was explored through the PES framework to overcome inter-individual heterogeneity and enable precision drug repurposing in RA. The results showed a statistically significant association between RA polygenic scores and self-reported RA and individual PES scorings for 3,719 RA patients. Interestingly, several enriched PES pathways were targeted by already approved RA drugs. In addition, the analysis revealed genetically supported drug repurposing opportunities for future treatment of RA with a relatively safe profile.

Keywords: rheumatoid arthritis, precision medicine, drug repurposing, system biology, bioinformatics

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Authors: Kyoung Lee

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Numerous aerobic bacteria have the ability to form multicellular communities on the surface layer of the air-liquid (A-L) interface as a biofilm called a pellicle. Pellicles occupied at the A-L interface will benefit from the utilization of oxygen from air and nutrient from liquid. Buoyancy of cells can be obtained by high surface tension at the A-L interface. Thus, formation of pellicles is an adaptive advantage in utilization of excess nutrients in the standing culture where oxygen depletion is easily set up due to rapid cell growth. In natural environments, pellicles are commonly observed on the surface of lake or pond contaminated with pollutants. Previously, we have shown that when cultured in standing LB media an alkylphenol-degrading bacteria Pseudomonas alkylphenolia KL28 forms pellicles in a diameter of 0.3-0.5 mm with a thickness of ca 40 µm. The pellicles have unique features for possessing flatness and unusual rigidity. In this study, the biogenesis of the circular pellicles has been investigated by observing the cell organization at early stages of pellicle formation and cell arrangements in pellicle, providing a clue for highly organized cellular arrangement to be adapted to the air-liquid niche. Here, we first monitored developmental patterns of pellicle from monolayer to multicellular organization. Pellicles were shaped by controlled growth of constituent cells which accumulate extracellular polymeric substance. The initial two-dimensional growth was transited to multilayers by a constraint force of accumulated self-produced extracellular polymeric substance. Experiments showed that pellicles are formed by clonal growth and even with knock-out of genes for flagella and pilus formation. In contrast, the mutants in the epm gene cluster for alginate-like polymer biosynthesis were incompetent in cell alignment for initial two-dimensional growth of pellicles. Electron microscopic and confocal laser scanning microscopic studies showed that the fully matured structures are highly packed by matrix-encased cells which have special arrangements. The cells on the surface of the pellicle lie relatively flat and inside longitudinally cross packed. HPLC analysis of the extrapolysaccharide (EPS) hydrolysate from the colonies from LB agar showed a composition with L-fucose, L-rhamnose, D-galactosamine, D-glucosamine, D-galactose, D-glucose, D-mannose. However, that from pellicles showed similar neutral and amino sugar profile but missing galactose. Furthermore, uronic acid analysis of EPS hydrolysates by HPLC showed that mannuronic acid was detected from pellicles not from colonies, indicating the epm-derived polymer is critical for pellicle formation as proved by the epm mutants. This study verified that for the circular pellicle architecture P. alkylphenolica KL28 cells utilized EPS building blocks different from that used for colony construction. These results indicate that P. alkylphenolica KL28 is a clever architect that dictates unique cell arrangements with selected EPS matrix material to construct sophisticated building, circular biofilm pellicles.

Keywords: biofilm, matrix, pellicle, pseudomonas

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Authors: Muhammad Tariq, Muhammad Waseem, Muhammad Hidayat Rasool

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Isolation and characterization of chromium tolerant Staphylococcus aureus from industrial wastewater and their potential use to bioremediate environmental chromium. Objectives: Chromium with its great economic importance in industrial use is major metal pollutant of the environment. Chromium are used in different industries for various applications such as textile, dyeing and pigmentation, wood preservation, manufacturing pulp and paper, chrome plating, steel and tanning. The release of untreated chromium in industrial effluents causes serious threat to environment and human health, therefore, the current study designed to isolate chromium tolerant Staphylococcus aureus for removal of chromium prior to their final discharge into the environment due to its cost effective and beneficial advantage over physical and chemical methods. Methods: Wastewater samples were collected from discharge point of different industries. Heavy metal analysis by atomic absorption spectrophotometer and microbiological analysis such as total viable count, total coliform, fecal coliform and Escherichia coli were conducted. Staphylococcus aureus was identified through gram’s staining, biomeriux vitek 2 microbial identification system and 16S rRNA gene amplification by polymerase chain reaction. Optimum growth conditions with respect to temperature, pH, salt concentrations and effect of chromium on the growth of bacteria, resistance to other heavy metal ions, minimum inhibitory concentration and chromium uptake ability of Staphylococcus aureus strain K1 was determined by spectrophotometer. Antibiotic sensitivity pattern was also determined by disc diffusion method. Furthermore, chromium uptake ability was confirmed by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope equipped with Oxford Energy Dipersive X-ray (EDX) micro analysis system. Results: The results presented that optimum temperature was 35ᵒC, pH was 8.0 and salt concentration was 0.5% for growth of Staphylococcus aureus K1. The maximum uptake ability of chromium by bacteria was 20mM than other heavy metal ions. The antibiotic sensitivity pattern revealed that Staphylococcus aureus was vancomycin and methicillin sensitive. Non hemolytic activity on blood agar and negative coagulase reaction showed that it was non-pathogenic. Furthermore, the growth of bacteria decreases in the presence of chromium and maximum chromium uptake by bacteria observed at optimum growth conditions. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and Energy dispersive X-ray (EDX) analysis confirmed the presence of chromium uptake by Staphylococcus aureus K1. Conclusion: The study revealed that Staphylococcus aureus K1 have the potential to bio-remediate chromium toxicity from wastewater. Gradually, this biological treatment becomes more important due to its advantage over physical and chemical methods to protect environment and human health.

Keywords: wastewater, staphylococcus, chromium, bioremediation

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Authors: Wang Hong, Liu Chang Kui, Zhao Bing Jing, Hu Min

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Objection We observed the repairment effection of canine mandibular defect with meshy Ti6Al4V scaffold fabricated by electron beam melting (EBM) combined with bone marrow mesenchymal stem cells (BMMSCs) encapsulated in chitosan hydrogel. Method Meshy titanium scaffolds were prepared by EBM of commercial Ti6Al4V power. The length of scaffolds was 24 mm, the width was 5 mm and height was 8mm. The pore size and porosity were evaluated by scanning electron microscopy (SEM). Chitosan /Bio-Oss hydrogel was prepared by chitosan, β- sodium glycerophosphate and Bio-Oss power. BMMSCs were harvested from canine iliac crests. BMMSCs were seeded in titanium scaffolds and encapsulated in Chitosan /Bio-Oss hydrogel. The validity of BMMSCs was evaluated by cell count kit-8 (CCK-8). The osteogenic differentiation ability was evaluated by alkaline phosphatase (ALP) activity and gene expression of OC, OPN and CoⅠ. Combination were performed by injecting BMMSCs/ Chitosan /Bio-Oss hydrogel into the meshy Ti6Al4V scaffolds and solidified. 24 mm long box-shaped bone defects were made at the mid-portion of mandible of adult beagles. The defects were randomly filled with BMMSCs/ Chitosan/Bio-Oss + titanium, Chitosan /Bio-Oss+titanium, titanium alone. Autogenous iliac crests graft as control group in 3 beagles. Radionuclide bone imaging was used to monitor the new bone tissue at 2, 4, 8 and 12 weeks after surgery. CT examination was made on the surgery day and 4 weeks, 12 weeks and 24 weeks after surgery. The animals were sacrificed in 4, 12 and 24 weeks after surgery. The bone formation were evaluated by histology and micro-CT. Results: The pores of the scaffolds was interconnected, the pore size was about 1 mm, the average porosity was about 76%. The pore size of the hydrogel was 50-200μm and the average porosity was approximately 90%. The hydrogel were solidified under the condition of 37℃in 10 minutes. The validity and the osteogenic differentiation ability of BMSCs were not affected by titanium scaffolds and hydrogel. Radionuclide bone imaging shown an increasing tendency of the revascularization and bone regeneration was observed in all the groups at 2, 4, 8 weeks after operation, and there were no changes at 12weeks.The tendency was more obvious in the BMMSCs/ Chitosan/Bio-Oss +titanium group and autogenous group. CT, Micro-CT and histology shown that new bone formed increasingly with the time extend. There were more new bone regenerated in BMMSCs/ Chitosan /Bio-Oss + titanium group and autogenous group than the other two groups. At 24 weeks, the autogenous group was achieved bone union. The BMSCs/ Chitosan /Bio-Oss group was seen extensive new bone formed around the scaffolds and more new bone inside of the central pores of scaffolds than Chitosan /Bio-Oss + titanium group and titanium group. The difference was significantly. Conclusion: The titanium scaffolds fabricated by EBM had controlled porous structure, good bone conduction and biocompatibility. Chitosan /Bio-Oss hydrogel had injectable plasticity, thermosensitive property and good biocompatibility. The meshy Ti6Al4V scaffold produced by EBM combined BMSCs encapsulated in chitosan hydrogel had good capacity on mandibular bone defect repair.

Keywords: mandibular reconstruction, tissue engineering, electron beam melting, titanium alloy

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Authors: Ram Mohan, Mahendran Samykano, Shyam Aravamudhan

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Metallic nanowires with sub-micron and hundreds of nanometer diameter have a diversity of applications in nano/micro-electromechanical systems (NEMS/MEMS). Characterizing the mechanical properties of such sub-micron and nano-scale metallic nanowires are tedious; require sophisticated and careful experimentation to be performed within high-powered microscopy systems (scanning electron microscope (SEM), atomic force microscope (AFM)). Also, needed are nanoscale devices for placing the nanowires; loading them with the intended conditions; obtaining the data for load–deflection during the deformation within the high-powered microscopy environment poses significant challenges. Even picking the grown nanowires and placing them correctly within a nanoscale loading device is not an easy task. Mechanical characterizations through experimental methods for such nanowires are still very limited. Various techniques at different levels of fidelity, resolution, and induced errors have been attempted by material science and nanomaterial researchers. The methods for determining the load, deflection within the nanoscale devices also pose a significant problem. The state of the art is thus still at its infancy. All these factors result and is seen in the wide differences in the characterization curves and the reported properties in the current literature. In this paper, we discuss and present our experimental method, results, and discussions of uniaxial tensile loading and the development of subsequent stress–strain characteristics curves for Nickel nanowires. Nickel nanowires in the diameter range of 220–270 nm were obtained in our laboratory via an electrodeposition method, which is a solution based, template method followed in our present work for growing 1-D Nickel nanowires. Process variables such as the presence of magnetic field, its intensity; and varying electrical current density during the electrodeposition process were found to influence the morphological and physical characteristics including crystal orientation, size of the grown nanowires1. To further understand the correlation and influence of electrodeposition process variables, associated formed structural features of our grown Nickel nanowires to their mechanical properties, careful experiments within scanning electron microscope (SEM) were conducted. Details of the uniaxial tensile characterization, testing methodology, nanoscale testing device, load–deflection characteristics, microscopy images of failure progression, and the subsequent stress–strain curves are discussed and presented.

Keywords: uniaxial tensile characterization, nanowires, electrodeposition, stress-strain, nickel

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Authors: Gurjeet Singh, Harinder Singh

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In this paper, we present "The Language of Science in Higher Education: Related Questions and Discussions". Linguists have written and researched in depth the role of language in science. On this basis, it is clear that language is not just a medium or vehicle for communicating knowledge and ideas. Nor are there mere signs of language knowledge and conversion of ideas into code. In the process of reading and writing, everyone thinks deeply and struggles to understand concepts and make sense. Linguistics play an important role in achieving concepts. In the context of such linguistic diversity, there is no straightforward and simple answer to the question of which language should be the language of advanced science and technology. Many important topics related to this issue are as follows: Involvement in practical or Deep theoretical issues. Languages for the study of science and other subjects. Language issues of science to be considered separate from the development of science, capitalism, colonial history, the worldview of the common man. The democratization of science and technology education in India is possible only by providing maximum reading/resource material in regional languages. The scientific research should be increase to chances of understanding the subject. Multilingual instead or monolingual. As far as deepening the understanding of the subject is concerned, we can shed light on it based on two or three experiences. An attempt was made to make the famous sociological journal Economic and Political Weekly Hindi almost three decades ago. There were many obstacles in this work. The original articles written in Hindi were not found, and the papers and articles of the English Journal were translated into Hindi, and a journal called Sancha was taken out. Equally important is the democratization of knowledge and the deepening of understanding of the subject. However, the question is that if higher education in science is in Hindi or other languages, then it would be a problem to get job. In fact, since independence, English has been dominant in almost every field except literature. There are historical reasons for this, which cannot be reversed. As mentioned above, due to colonial rule, even before independence, English was established as a language of communication, the language of power/status, the language of higher education, the language of administration, and the language of scholarly discourse. After independence, attempts to make Hindi or Hindustani the national language in India were unsuccessful. Given this history and current reality, higher education should be multilingual or at least bilingual. Translation limits should also be increased for those who choose the material for translation. Writing in regional languages on science, making knowledge of various international languages available in Indian languages, etc., is equally important for all to have opportunities to learn English.

Keywords: language, linguistics, literature, culture, ethnography, punjabi, gurmukhi, higher education

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Authors: Nivedhitha Venkatakrishnan

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Walking down busy temple streets in India is an experience in lifetime. Especially the temple streets are one of the most energetic places not only because of the divinity but also because of the streets itself which provides place for people to relax, meet, shop, linger, just walk around these activities create a set of experience which results in memories that lasts longer. Thinking of any temple street in India the image that comes to anyone’s mind are the elegantly sculpted Gopurams (Gateway) that depicts the craftsmanship and the history of the place, people taking a holy dip in the water, the aroma of the agarbathi’s, flowers with the divine Vedic chants and the sound of the temple bell flock of pigeons flying from the niches of the Gopuram with the sun in the backdrop. It gives a feeling of impulse energy that brings in life to these streets. Any temple street with even any one factor missing would look dead. This will be amiss in the essence in the scene of one’s experiences. These Temple Streets traditionally cater not only for religious purpose but to a wide range of activities. A vibrant street that facilitates such activities are preferred by the public any day. The research seeks to understand and find out the definition of Vibrancy in Indian Context. What is Vibrancy? What brings in the feeling of Vibrancy/Liveliness/Energy? Is it the Built structure and the city? Or is it the people? Or is it the Activity? Or is it Built structure – city – People – Activity put together brings the sense of Vibrancy to a place? How to define Vibrancy? Is it measurable? For which a case of Car Street Udupi, Karnataka is taken. The research is carried out in two stages. ‘Stage One’ makes use of ethnographic fieldwork as a basic method, complimented by structured field observations using a behavioral mapping procedure of the streets. Stage Two’ utilizes surveys that collected. This stage seeks to understand what design characteristics and furniture arrangements are associated with stationary, social and gathering activities of people by each cultural group and all groups collectively. The main conclusion from this research is that retail activities remain the main concern of people in cultural streets. Management and higher-level planning of retail activities on the streets could encourage and motivate possible Shops to enrich the trade variety of the street that provides a means for social and cultural diversity. In addition to business activities, spatial design characteristics are found to have an influence on people’s behavior and activity. The findings of this research suggest that retail and business activities, together with the design and skillful management of the public areas, could support a wider range of static and social activities among people of various ethnic backgrounds.

Keywords: activity, liveliness, temple street, vibrancy

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Authors: Austin Vander Wel, Katherin Vargas Henao

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

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

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Authors: Daniel Todorov, Anton Hinkov, Petya Angelova, Kalina Shishkova, Venelin Tsvetkov, Stoyan Shishkov

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Human herpes viruses (HHV) are ubiquitous pathogens with a pandemic spread across the globe. HHV type 1 is the main causative agent of cold sores and fever blisters around the mouth and on the face, whereas HHV type 2 is generally responsible for genital herpes outbreaks. The treatment of both viruses is more or less successful with antivirals from the nucleoside analogues group. Their wide application increasingly leads to the emergence of resistant mutants In the past, medicinal plants have been used to treat a number of infectious and non-infectious diseases. Their diversity and ability to produce the vast variety of secondary metabolites according to the characteristics of the environment give them the potential to help us in our warfare with viral infections. The variable chemical characteristics and complex composition is an advantage in the treatment of herpes since the emergence of resistant mutants is significantly complicated. The screening process is difficult due to the lack of standardization. That is why it is especially important to follow the mechanism of antiviral action of plants. On the one hand, it may be expected to interact with its compounds, resulting in enhanced antiviral effects, and the most appropriate environmental conditions can be chosen to maximize the amount of active secondary metabolites. During our study, we followed the activity of various plant extracts on the viral replication cycle as well as their effect on the extracellular virion. We obtained our results following the logical sequence of the experimental settings - determining the cytotoxicity of the extracts, evaluating the overall effect on viral replication and extracellular virion.During our research, we have screened a variety of plant extracts for their antiviral activity against both virus replication and the virion itself. We investigated the effect of the extracts on the individual stages of the viral replication cycle - viral adsorption, penetration and the effect on replication depending on the time of addition. If there are positive results in the later experiments, we had studied the activity over viral adsorption, penetration and the effect of replication according to the time of addition. Our results indicate that some of the extracts from the Lamium album have several targets. The first stages of the viral life cycle are most affected. Several of our active antiviral agents have shown an effect on extracellular virion and adsorption and penetration processes. Our research over the last decade has shown several curative antiviral plants - some of which are from the Lamiacea family. The rich set of active ingredients of the plants in this family makes them a good source of antiviral preparation.

Keywords: human herpes virus, antiviral activity, Lamium album, Nepeta nuda

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Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino

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Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.

Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus

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Authors: Bhargavi G. Iyer, Ojaswi Bhagat

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In recent years, there has been a lot of discussion about ethnic imbalance and diversity in the international context. Arbitration is now subject to the hegemony of a small number of people who are constantly reappointed. When a court system becomes exclusionary, the quality of adjudication suffers significantly. In such a framework, there is a misalignment between adjudicators' preconceived views and the interests of the parties, resulting in a biased view of the proceedings. The world is currently witnessing a slew of intellectual property battles around cultural appropriation. The term "cultural appropriation" refers to the industrial west's theft of indigenous culture, usually for fashion, aesthetic, or dramatic purposes. Selena Gomez exemplifies cultural appropriation by commercially using the “bindi,” which is sacred to Hinduism, as a fashion symbol. In another case, Victoria's Secret insulted indigenous peoples' genocide by stealing native Indian headdresses. In the case of yoga, a similar process can be witnessed, with Vedic philosophy being reduced to a type of physical practice. Such a viewpoint is problematic since indigenous groups have worked hard for generations to ensure the survival of their culture, and its appropriation by the western world for purely aesthetic and theatrical purposes is upsetting to those who practise such cultures. Because such conflicts involve numerous jurisdictions, they must be resolved through international arbitration. However, these conflicts are already being litigated, and the aggrieved parties, namely developing nations, do not believe it prudent to use the World Intellectual Property Organization's (WIPO) already established arbitration procedure. This practise, it is suggested in this study, is the outcome of Europe's exclusionary arbitral system, which fails to recognise the non-legal and non-commercial nature of indigenous culture issues. This research paper proposes a more comprehensive, inclusive approach that recognises the non-legal and non-commercial aspects of IP disputes involving cultural appropriation, which can only be achieved through an ethnically balanced arbitration structure. This paper also aspires to expound upon the benefits of arbitration and other means of alternative dispute resolution (ADR) in the context of disputes pertaining to cultural issues; positing that inclusivity is a solution to the existing discord between international practices and localised cultural points of dispute. This paper also hopes to explicate measures that will facilitate ensuring inclusion and ideal practices in the domain of arbitration law, particularly pertaining to cultural heritage and indigenous expression.

Keywords: arbitration law, cultural appropriation, dispute resolution, heritage, intellectual property

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Authors: Doreen Mekunda

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This paper is built on the premise that the current socio-political crisis in the two restive regions of Cameroon, though enveloped with destructive and devastating trends (effects) on both property and human lives, is not without its strengths and merits. It is incontestable that many cultures, to a greater extent, are going to be destroyed as people forcibly move from war-stricken habitats to non-violent places. Many cultural potentials, traditional shrines, artifacts, art, and crafts, etc., are unknowingly or knowingly disfigured, and many other ugly things will, by the end of the crisis, affect the cultures of these two regions under siege and of the receiving population. A plethora of other problems like the persecution of Internally Displaced Persons (IDPs) for being displaced and blamed for increased crime rates and the existence of cultural and ethnic differences that produce both inter-tribal and interpersonal conflicts and conflicts between communities will abound. However, there is the emergence of rapid literature, and other forms of cultural productions, whether written or oral, is visible, thereby precipitating a rich cultural diversity due to the coming together of a variety of cultures of both the IDPs and the receiving populations, rapid urbanization, improvement of health-related issues, the rebirth of indigenous cultural practices, the development of social and lingua-cultural competences, dependence on alternative religions, faith and spirituality. Even financial and economic dependence, though a burden to others by IDPs, has its own merits as it improves the living standards of the IDPs. To be able to obtain plausible results, cultural materialism, which is a literary theory that hinges on the empirical study of socio-cultural systems within a materialist infrastructure-super-structure framework, is employed together with the postcolonial theory. Postcolonial theory because the study deals with postcolonial experiences/tenets of migration, hybridity, ethnicity, indignity, language, double consciousness, migration, center/margin binaries, and identity, amongst others. The study reveals that the involuntary movement of persons from their habitual homes brings about movement in cultures, thus, the emergence of new cultures. The movement of people who hold fast to their cultural heritage can only influence new forms of literature, the development of new communication competences, the rise of alternative religion, faith and spirituality, the re-emergence of customary and traditional legal systems that might have been abandoned for the new judicial systems, and above all the revitalization of traditional health care systems.

Keywords: alternative religion, emergence, socio-political crisis, spirituality, lingua-cultural competences

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Authors: Mohamad Chour

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Nowadays, a record number of households are headed by single fathers in most of the European societies. Our research aims to explore how French single fathers experience the domestic sphere, a traditionally feminized field while accomplishing their role of fathers. We adopt a gender role and a parenting role construction theoretical perspectives. Indeed, the interior domestic sphere has been traditionally considered as related to the role of the mother. Moreover, according to the masculine domination theory of Bourdieu, men avoid caregiving and domestic practices that are economically and culturally undervalued. Hence, mothers are considered as more likely to handle the expressive dimension of duties whereas fathers’ role is represented as instrumental, functional and independent. Long interviews have been conducted with twenty French single fathers in order to investigate how the absence of the mother affects the practices of fatherhood. We combined the long interviews with projective techniques method in order to better understand their conception of the family and their family values. Seeking a qualitative diversity, our respondents are from various ages (between 30 and 60); they are coming from different regions in France; living in rural, semi-rural and urban areas. Based on the analysis of 427 pages of data, we identify three main categories of single fathers depending on their strategies to copy and/or delegate the role of the mother. 1) Nurturing fathers completely assume the role of the absent mother as well as her functions. Their discourse is characterized by abnegation and sacrifices reflecting a nurturing role. 2) Juggling fathers are those who take charge of a part of the household duties and delegate the other part to the market or to 'feminine resources' for lacking skills or time. 3) Resistant fathers are the very few respondents who refuse to assume any activities related to the domestic sphere that they perceive as feminine. For lacking competences and even for ideological reasons, they have tendency to delegate all the tasks that were assumed by their ex-spouses. Generally, the majority of fathers seem to experience the domestic sphere differently, and their domestic involvement has been underestimated and even misunderstood. Household duties such as cooking and housekeeping in addition to the nurturing role are experienced by many of the respondents as constructing elements of their fatherhood. Our respondents do not seem to accomplish house holding duties in a functional way. The domestic sphere is managed by those fathers with a strong dimension of abnegation. Thus, our research contributes to illustrating the evolution of gender roles and shows how being simultaneously 'a father and a mother' seems to be an emerging social norm in a French and European cultural context.

Keywords: fathering, gender roles, gender studies, identity construction, single fathers

Procedia PDF Downloads 117

Authors: Chantal Maille

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This paper is about gender-based analysis plus (GBA+), an intersectional gender policy used in Canada to assess the impact of policies and programs for men and women from different origins. It looks at Canada’s political culture to explain the nature of its gender policies. GBA+ is defined as an analysis method that makes it possible to assess the eventual effects of policies, programs, services, and other initiatives on women and men of different backgrounds because it takes account of gender and other identity factors. The ‘plus’ in the name serves to emphasize that GBA+ goes beyond gender to include an examination of a wide range of other related identity factors, such as age, education, language, geography, culture, and income. The point of departure for GBA+ is that women and men are not homogeneous populations and gender is never the only factor in defining a person’s identity; rather, it interacts with factors such as ethnic origin, age, disabilities, where the person lives, and other aspects of individual and social identity. GBA+ takes account of these factors and thus challenges notions of similarity or homogeneity within populations of women and men. Comparative analysis based on sex and gender may serve as a gateway to studying a given question, but women, men, girls, and boys do not form homogeneous populations. In the 1990s, intersectionality emerged as a new feminist framework. The popularity of the notion of intersectionality corresponds to a time when, in hindsight, the damage done to minoritized groups by state disengagement policies in concert with global intensification of neoliberalism, and vice versa, can be measured. Although GBA+ constitutes a form of intersectionalization of GBA, it must be understood that the two frameworks do not spring from a similar logic. Intersectionality first emerged as a dynamic analysis of differences between women that was oriented toward change and social justice, whereas GBA is a technique developed by state feminists in a context of analyzing governmental policies and aiming to promote equality between men and women. It can nevertheless be assumed that there might be interest in such a policy and program analysis grid that is decentred from gender and offers enough flexibility to take account of a group of inequalities. In terms of methodology, the research is supported by a qualitative analysis of governmental documents about GBA+ in Canada. Research findings identify links between Canadian gender policies and its political culture. In Canada, diversity has been taken into account as an element at the basis of gendered analysis of public policies since 1995. The GBA+ adopted by the government of Canada conveys an opening to intersectionality and a sensitivity to multiculturalism. The Canadian Multiculturalism Act, adopted 1988, proposes to recognize the fact that multiculturalism is a fundamental characteristic of the Canadian identity and heritage and constitutes an invaluable resource for the future of the country. In conclusion, Canada’s distinct political culture can be associated with the specific nature of its gender policies.

Keywords: Canada, gender-based analysis, gender policies, political culture

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Authors: Veronica Botero, Karen Ortiz

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The Faculty of Mines, of the National University of Colombia, Medellín Campus, is a faculty that has 136 years of history and represents one of the most important study centers in the country in the field of engineering and scientific research, as well as a reference at a global, national, and Latin American level in this matter. Despite being a faculty with so many years of history and having trained a large number of graduates under the traditional mechanistic and androcentric paradigm, which reproduces the logic of the traditional scientific method and the differentiated and severe look between subject-object of research among other binarisms, has also been the place where professors and students have become aware of the need to transform this paradigm into engineering, and focus on the sustainability of diversity and the well-being of the natural and social systems that inhabit the territories and has opened possibilities for the implementation of classes that address feminist pedagogical theories and practices. The class: The return of the witches, is an initiative that constitutes an important training exercise that provides students with the study of feminisms, the importance of closing gender gaps and critical readings on the traditional paradigm of engineering. The objective of this article is to present a systematization of the experience of design, implementation and development of this elective class, describing the tensions that arose at the time when a subject of this style was created and proposed in the Department of Geosciences and Environment, from the Faculty of Mines in 2022; the reactions of the groups of students who have taken it and their perceptions and opinions about ecofeminism as proposals for critical analysis and practices in relation to the environment and, above all, how their readings of the world have changed after having studied this subject for a semester. The pedagogical journey and the feminist methodologies that have been designed and adjusted over two years of work will be explained based on the sharing of situated knowledge of the students and the two teachers who teach the course, who pose challenges to the dominant ideology in engineering since one of them is trained in human sciences and feminist studies and the other, although trained in civil engineering and geosciences, is a woman with diverse sexual orientation and is the first professor to have assumed the position of dean in the 135 years of history of the Faculty. The transformations in the life experience of the students are revealing since they affirm that the training process is forceful and powerful to outline a much more qualified and critical professional profile that contributes to the transformation of gender gaps in the country. This class is therefore a challenge in this Faculty of Engineering that still presents a dominant ideology on gender that has not been questioned or challenged before.

Keywords: feminisms, gender equality, gender bias, engineering for life Manifiesto.

Procedia PDF Downloads 41

Authors: G. Cunningham, E. Cantor, X. Wu, F. Shen, G. Jiang, S. Philips, C. Bales, Y. Xiao, T. R. Cummins, J. C. Fehrenbacher, B. P. Schneider

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Background: Taxane-induced peripheral neuropathy (TIPN) is the most devastating survivorship issue for patients receiving therapy. Dose reductions due to TIPN in the curative setting lead to inferior outcomes for African American patients, as prior research has shown that this group is more susceptible to developing severe neuropathy. The mechanistic underpinnings of TIPN, however, have not been entirely elucidated. While it would be appealing to use primary tissue to study the development of TIPN, procuring nerves from patients is not realistically feasible, as nerve biopsies are painful and may result in permanent damage. Therefore, our laboratory has investigated paclitaxel-induced neuronal morphological and molecular changes using an ex vivo model of human-induced pluripotent stem cell (iPSC)-derived neurons. Methods: iPSCs are undifferentiated and endlessly dividing cells that can be generated from a patient’s somatic cells, such as peripheral blood mononuclear cells (PBMCs). We successfully reprogrammed PBMCs into iPSCs using the Erythroid Progenitor Reprograming Kit (STEMCell Technologiesᵀᴹ); pluripotency was verified by flow cytometry analysis. iPSCs were then induced into neurons using a differentiation protocol that bypasses the neural progenitor stage and uses selected small-molecule modulators of key signaling pathways (SMAD, Notch, FGFR1 inhibition, and Wnt activation). Results: Flow cytometry analysis revealed expression of core pluripotency transcription factors Nanog, Oct3/4 and Sox2 in iPSCs overlaps with commercially purchased pluripotent cell line UCSD064i-20-2. Trilineage differentiation of iPSCs was confirmed with immunofluorescent imaging with germ-layer-specific markers; Sox17 and ExoA2 for ectoderm, Nestin, and Pax6 for mesoderm, and Ncam and Brachyury for endoderm. Sensory neuron markers, β-III tubulin, and Peripherin were applied to stain the cells for the maturity of iPSC-derived neurons. Patch-clamp electrophysiology and calcitonin gene-related peptide (CGRP) release data supported the functionality of the induced neurons and provided insight into the timing for which downstream assays could be performed (week 4 post-induction). We have also performed a cell viability assay and fluorescence-activated cell sorting (FACS) using four cell-surface markers (CD184, CD44, CD15, and CD24) to select a neuronal population. At least 70% of the cells were viable in the isolated neuron population. Conclusion: We have found that these iPSC-derived neurons recapitulate mature neuronal phenotypes and demonstrate functionality. Thus, this represents a patient-derived ex vivo neuronal model to investigate the molecular mechanisms of clinical TIPN.

Keywords: chemotherapy, iPSC-derived neurons, peripheral neuropathy, taxane, paclitaxel

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Authors: Orit Baruth, Anat Cohen

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Online courses have become common in many learning programs and various learning environments, particularly in higher education. Social distancing forced in response to the COVID-19 pandemic has increased the demand for these courses. Yet, despite the frequency of use, online learning is not free of limitations and may not suit all learners. Hence, the growth of online learning alongside with learners' diversity raises the question: is online learning, as it currently offered, meets the needs of each learner? Fortunately, today's technology allows to produce tailored learning platforms, namely, personalization. Personality influences learner's satisfaction and therefore has a significant impact on learning effectiveness. A better understanding of personality can lead to a greater appreciation of learning needs, as well to assists educators ensure that an optimal learning environment is provided. In the context of online learning and personality, the research on learning design according to personality traits is lacking. This study explores the relations between personality traits (using the 'Big-five' model) and students' satisfaction with five techno-pedagogical learning solutions (TPLS): discussion groups, digital books, online assignments, surveys/polls, and media, in order to provide an online learning process to students' satisfaction. Satisfaction level and personality identification of 108 students who participated in a fully online learning course at a large, accredited university were measured. Cluster analysis methods (k-mean) were applied to identify learners’ clusters according to their personality traits. Correlation analysis was performed to examine the relations between the obtained clusters and satisfaction with the offered TPLS. Findings suggest that learners associated with the 'Neurotic' cluster showed low satisfaction with all TPLS compared to learners associated with the 'Non-neurotics' cluster. learners associated with the 'Consciences' cluster were satisfied with all TPLS except discussion groups, and those in the 'Open-Extroverts' cluster were satisfied with assignments and media. All clusters except 'Neurotic' were highly satisfied with the online course in general. According to the findings, dividing learners into four clusters based on personality traits may help define tailor learning paths for them, combining various TPLS to increase their satisfaction. As personality has a set of traits, several TPLS may be offered in each learning path. For the neurotics, however, an extended selection may suit more, or alternatively offering them the TPLS they less dislike. Study findings clearly indicate that personality plays a significant role in a learner's satisfaction level. Consequently, personality traits should be considered when designing personalized learning activities. The current research seeks to bridge the theoretical gap in this specific research area. Establishing the assumption that different personalities need different learning solutions may contribute towards a better design of online courses, leaving no learner behind, whether he\ she likes online learning or not, since different personalities need different learning solutions.

Keywords: online learning, personality traits, personalization, techno-pedagogical learning solutions

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Authors: Ahmad Haidar

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The rapid adoption of Artificial Intelligence (AI) technology holds unforeseen risks like privacy violation, unemployment, and algorithmic bias, triggering research institutions, governments, and companies to develop principles of AI ethics. The extensive and diverse literature on AI lacks an analysis of the evolution of principles developed in recent years. There are two fundamental purposes of this paper. The first is to provide insights into how the principles of AI ethics have been changed recently, including concepts like risk management and public participation. In doing so, a NOISE (Needs, Opportunities, Improvements, Strengths, & Exceptions) analysis will be presented. Second, offering a framework for building Ethical AI linked to sustainability. This research adopts an explorative approach, more specifically, an inductive approach to address the theoretical gap. Consequently, this paper tracks the different efforts to have “trustworthy AI” and “ethical AI,” concluding a list of 12 documents released from 2017 to 2022. The analysis of this list unifies the different approaches toward trustworthy AI in two steps. First, splitting the principles into two categories, technical and net benefit, and second, testing the frequency of each principle, providing the different technical principles that may be useful for stakeholders considering the lifecycle of AI, or what is known as sustainable AI. Sustainable AI is the third wave of AI ethics and a movement to drive change throughout the entire lifecycle of AI products (i.e., idea generation, training, re-tuning, implementation, and governance) in the direction of greater ecological integrity and social fairness. In this vein, results suggest transparency, privacy, fairness, safety, autonomy, and accountability as recommended technical principles to include in the lifecycle of AI. Another contribution is to capture the different basis that aid the process of AI for sustainability (e.g., towards sustainable development goals). The results indicate data governance, do no harm, human well-being, and risk management as crucial AI for sustainability principles. This study’s last contribution clarifies how the principles evolved. To illustrate, in 2018, the Montreal declaration mentioned eight principles well-being, autonomy, privacy, solidarity, democratic participation, equity, and diversity. In 2021, notions emerged from the European Commission proposal, including public trust, public participation, scientific integrity, risk assessment, flexibility, benefit and cost, and interagency coordination. The study design will strengthen the validity of previous studies. Yet, we advance knowledge in trustworthy AI by considering recent documents, linking principles with sustainable AI and AI for sustainability, and shedding light on the evolution of guidelines over time.

Keywords: artificial intelligence, AI for sustainability, declarations, framework, regulations, risks, sustainable AI

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Authors: Ricardo Godoy, Herbert Venthur, Hector Jimenez, Andres Quiroz, Ana Mutis

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In agriculture, grape production has great economic importance at global level, considering that in 2013 it reached 7.4 million hectares (ha) covered by plantations of this fruit worldwide. Chile is the number one exporter in the world with 800,000 tons. However, these values have been threatened by the attack of the grapevine moth, Lobesia botrana (Denis & Schiffermuller) (Lepidoptera: Tortricidae), since its detection in 2008. Nowadays, the use of semiochemicals, in particular the major component of the sex pheromone, (E,Z)-7.9-dodecadienil acetate, are part of mating disruption methods to control L. botrana. How insect pests can recognize these molecules, is being part of huge efforts to deorphanize their olfactory mechanism at molecular level. Thus, an interesting group of proteins has been identified in the antennae of insects, where odorant-binding proteins (OBPs) are known by transporting molecules to odorant receptors (ORs) and a co-receptor (ORCO) causing a behavioral change in the insect. Other proteins such as chemosensory proteins (CSPs), ionotropic receptors (IRs), odorant degrading enzymes (ODEs) and sensory neuron membrane proteins (SNMPs) seem to be involved, but few studies have been performed so far. The above has led to an increasing interest in insect communication at a molecular level, which has contributed to both a better understanding of the olfaction process and the design of new pest management strategies. To date, it has been reported that the ORs can detect one or a small group of odorants in a specific way. Therefore, the objective of this study is the identification of genes that encode these ORs using the antennal transcriptome of L. botrana. Total RNA was extracted for females and males of L. botrana, and the antennal transcriptome sequenced by Next Generation Sequencing service using an Illumina HiSeq2500 platform with 50 million reads per sample. Unigenes were assembled using Trinity v2.4.0 package and transcript abundance was obtained using edgeR. Genes were identified using BLASTN and BLASTX locally installed in a Unix system and based on our own Tortricidae database. Those Unigenes related to ORs were characterized using ORFfinder and protein Blastp server. Finally, a phylogenetic analysis was performed with the candidate amino acid sequences for LbotORs including amino acid sequences of other moths ORs, such as Bombyx mori, Cydia pomonella, among others. Our findings suggest 61 genes encoding ORs and one gene encoding an ORCO in both sexes, where the greatest difference was found in the OR6 because of the transcript abundance according to the value of FPKM in females and males was 1.48 versus 324.00. In addition, according to phylogenetic analysis OR6 is closely related to OR1 in Cydia pomonella and OR6, OR7 in Epiphyas postvittana, which have been described as pheromonal receptors (PRs). These results represent the first evidence of ORs present in the antennae of L. botrana and a suitable starting point for further functional studies with selected ORs, such as OR6, which is potentially related to pheromonal recognition.

Keywords: antennal transcriptome, lobesia botrana, odorant receptors (ORs), phylogenetic analysis

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Authors: Gergely Horvath

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The study analyzes the special provisions of the highest level of national agricultural legislation in the Fundamental Law of Hungary (25 April 2011) with descriptive, analytic and comparative methods. The agriculturally relevant articles of the constitution are very important, because –in spite of their high level of abstraction– they can determine and serve the practice comprehensively and effectively. That is why the objective of the research is to interpret the concrete sentences and phrases in connection with agriculture compared with the methods of some other relevant constitutions (historical-grammatical interpretation). The major findings of the study focus on searching for the appropriate provisions and approach capable of solving the problems of sustainable food production. The real challenge agricultural law must face with in the future is protecting or conserving its background and subjects: the environment, the ecosystem services and all the 'roots' of food production. In effect, agricultural law is the legal aspect of the production of 'our daily bread' from farm to table. However, it also must guarantee the safe daily food for our children and for all our descendants. In connection with sustainability, this unique, value-oriented constitution of an agrarian country even deals with uncustomary questions in this level of legislation like GMOs (by banning the production of genetically modified crops). The starting point is that the principle of public good (principium boni communis) must be the leading notion of the norm, which is an idea partly outside the law. The public interest is reflected by the agricultural law mainly in the concept of public health (in connection with food security) and the security of supply with healthy food. The construed Article P claims the general protection of our natural resources as a requirement. The enumeration of the specific natural resources 'which all form part of the common national heritage' also means the conservation of the grounds of sustainable agriculture. The reference of the arable land represents the subfield of law of the protection of land (and soil conservation), that of the water resources represents the subfield of water protection, the reference of forests and the biological diversity visualize the specialty of nature conservation, which is an essential support for agrobiodiversity. The mentioned protected objects constituting the nation's common heritage metonymically melt with their protective regimes, strengthening them and forming constitutional references of law. This regimes also mean the protection of the natural foundations of the life of the living and also the future generations, in the name of intra- and intergenerational equity.

Keywords: agricultural law, constitutional values, natural resources, sustainability

Procedia PDF Downloads 147

Authors: Alena Cejkova, Martina Paldrychova, Jana Michailidu, Olga Matatkova, Jan Masak

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Increasing the resistance of pathogenic microorganisms to many antibiotics is a serious threat to the treatment of infectious diseases and cleaning medical instruments. It should be added that the resistance of microbial populations growing in biofilms is often up to 1000 times higher compared to planktonic cells. Biofilm formation in a number of microorganisms is largely influenced by the quorum sensing regulatory mechanism. Finding external factors such as natural substances or physical processes that can interfere effectively with quorum sensing signal molecules should reduce the ability of the cell population to form biofilm and increase the effectiveness of antibiotics. The present work is devoted to the effect of chitosan as a representative of natural substances with anti-biofilm activity and non- thermal plasma (NTP) alone or in combination with polymyxin B on biofilm formation of Pseudomonas aeruginosa. Particular attention was paid to the influence of these agents on the level of quorum sensing signal molecules (acyl-homoserine lactones) during planktonic and biofilm cultivations. Opportunistic pathogenic strains of Pseudomonas aeruginosa (DBM 3081, DBM 3777, ATCC 10145, ATCC 15442) were used as model microorganisms. Cultivations of planktonic and biofilm populations in 96-well microtiter plates on horizontal shaker were used for determination of antibiotic and anti-biofilm activity of chitosan and polymyxin B. Biofilm-growing cells on titanium alloy, which is used for preparation of joint replacement, were exposed to non-thermal plasma generated by cometary corona with a metallic grid for 15 and 30 minutes. Cultivation followed in fresh LB medium with or without chitosan or polymyxin B for next 24 h. Biofilms were quantified by crystal violet assay. Metabolic activity of the cells in biofilm was measured using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) colorimetric test based on the reduction of MTT into formazan by the dehydrogenase system of living cells. Activity of N-acyl homoserine lactones (AHLs) compounds involved in the regulation of biofilm formation was determined using Agrobacterium tumefaciens strain harboring a traG::lacZ/traR reporter gene responsive to AHLs. The experiments showed that both chitosan and non-thermal plasma reduce the AHLs level and thus the biofilm formation and stability. The effectiveness of both agents was somewhat strain dependent. During the eradication of P. aeruginosa DBM 3081 biofilm on titanium alloy induced by chitosan (45 mg / l) there was an 80% decrease in AHLs. Applying chitosan or NTP on the P. aeruginosa DBM 3777 biofilm did not cause a significant decrease in AHLs, however, in combination with both (chitosan 55 mg / l and NTP 30 min), resulted in a 70% decrease in AHLs. Combined application of NTP and polymyxin B allowed reduce antibiotic concentration to achieve the same level of AHLs inhibition in P. aeruginosa ATCC 15442. The results shown that non-thermal plasma and chitosan have considerable potential for the eradication of highly resistant P. aeruginosa biofilms, for example on medical instruments or joint implants.

Keywords: anti-biofilm activity, chitosan, non-thermal plasma, opportunistic pathogens

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Authors: Cazadira Fediva Tamzil, Agung Sulthonaulia Utama

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The rise in popularity of Indonesian Islamic fashion displayed and sold through social networking sites, especially Instagram, might seem at first glance like a commonplace and localized phenomenon. However, when analyzed critically, it actually reveals the relations between the global and local Indonesian economy, as well as a deep socio-anthropological dimension relating to religion, culture, class, work, identity. Conducted using a qualitative methodology, data collection technique of literature review, and observation of various social networking sites, this research finds four things that lead to the aforementioned conclusion. First, the rise of online Islamic fashion retailers was triggered by the shift in the structure of global and national Indonesian economy as well as the free access of information made possible by democratization in Indonesia and worldwide advances in terms of technology. All of those factors combined together gave birth to a large amount of middle-class Indonesians with high consumer culture and entrepreneurial flair. Second, online Islamic fashion retailers are the new cultural trendsetters in society. All these show how Indonesians are becoming increasingly pious, no longer only adhere to Western conception of luxury and that many are increasingly exploiting Islam commercial and status-acquiring purposes. Third, the online Islamic fashion retailers actually reveal a shift in the conception of ‘work’ – social media has made work no longer only confined to the toiling activities inside factories, but instead something that can be done from any location only through posting online words or pictures that can increase a fashion product’s capital value. Without realizing it, many celebrities and online retailers who promote Islamic fashion through social media on a daily basis are now also ‘semi-free immaterial labors’ – a slight reconceptualization to Tiziana Terranova’s concept of ‘free labor’ and Maurizio Lazzarato’s ‘immaterial labor’, which basically refer to people who create economic value and thus help out capitals from producing immaterial things with only little compensation in return. Fourth, this research also shows that the diversity of Islamic fashion styles being sold on Instagram reflects the polarized identity of Islam in Indonesia. In stark contrast with the theory which states that globalization always leads to the strengthening and unification of identity, this research shows how polarized the Islamic identity in Indonesia really is – even in the face of globalization.

Keywords: global economy, Indonesian online Islamic fashion, political relations, socio-anthropology

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Authors: Zoltan Sebestyen, Eszter Barta-Rajnai, Emma Jakab, Zsuzsanna Czegeny

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Extraction of the energy content of lignocellulosic biomass is one of the possible pathways to reduce the greenhouse gas emission derived from the burning of the fossil fuels. The application of the bioenergy can mitigate the energy dependency of a country from the foreign natural gas and the petroleum. The diversity of the plant materials makes difficult the utilization of the raw biomass in power plants. This problem can be overcome by the application of thermochemical techniques. Pyrolysis is the thermal decomposition of the raw materials under inert atmosphere at high temperatures, which produces pyrolysis gas, biooil and charcoal. The energy content of these products can be exploited by further utilization. The differences in the chemical and physical properties of the raw biomass materials can be reduced by the use of torrefaction. Torrefaction is a promising mild thermal pretreatment method performed at temperatures between 200 and 300 °C in an inert atmosphere. The goal of the pretreatment from a chemical point of view is the removal of water and the acidic groups of hemicelluloses or the whole hemicellulose fraction with minor degradation of cellulose and lignin in the biomass. Thus, the stability of biomass against biodegradation increases, while its energy density increases. The volume of the raw materials decreases so the expenses of the transportation and the storage are reduced as well. Biooil is the major product during pyrolysis and an important by-product during torrefaction of biomass. The composition of biooil mostly depends on the quality of the raw materials and the applied temperature. In this work, thermoanalytical techniques have been used to study the qualitative and quantitative composition of the pyrolysis and torrefaction oils of a woody (black locust) and two herbaceous samples (rape straw and wheat straw). The biooil contains C5 and C6 anhydrosugar molecules, as well as aromatic compounds originating from hemicellulose, cellulose, and lignin, respectively. In this study, special emphasis was placed on the formation of the lignin monomeric products. The structure of the lignin fraction is different in the wood and in the herbaceous plants. According to the thermoanalytical studies the decomposition of lignin starts above 200 °C and ends at about 500 °C. The lignin monomers are present among the components of the torrefaction oil even at relatively low temperatures. We established that the concentration and the composition of the lignin products vary significantly with the applied temperature indicating that different decomposition mechanisms dominate at low and high temperatures. The evolutions of decomposition products as well as the thermal stability of the samples were measured by thermogravimetry/mass spectrometry (TG/MS). The differences in the structure of the lignin products of woody and herbaceous samples were characterized by the method of pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS). As a statistical method, principal component analysis (PCA) has been used to find correlation between the composition of lignin products of the biooil and the applied temperatures.

Keywords: pyrolysis, torrefaction, biooil, lignin

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Authors: Rui Brito

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This contribution regards a literature review of inventory management that is a relevant topic for the firms, due to its important use of capital with implications in firm’s profitability within the complexity of a more competitive and globalized world. Firms look for small inventories in order to reduce holding costs, namely opportunity cost, warehousing and handling costs, deterioration and being out of style, but larger inventories are required by some reasons, such as customer service, ordering cost, transportation cost, supplier’s payment to reduce unit costs or to take advantage of price increase in the near future, and equipment setup cost. Thus, management shall address a trade-off between small inventories and larger inventories. This literature review concerns three types of inventory (spare parts, safety stock, and vendor) whose management usually is beyond the scope of logistics. The applied methodology consisted of an online search of databases regarding scientific documents in English, namely Elsevier, Springer, Emerald, Wiley, and Taylor & Francis, but excluding books except if edited, using search engines, such as Google Scholar and B-on. The search was based on three keywords/strings (themes) which had to be included just as in the article title, suggesting themes were very relevant to the researchers. The whole search period was between 2009 and 2018 with the aim of collecting between twenty and forty studies considered relevant within each of the key words/strings specified. Documents were sorted by relevance and to prevent the exclusion of the more recent articles, based on lower quantity of citations partially due to less time to be cited in new research articles, the search period was divided into two sub-periods (2009-2015 and 2016-2018). The number of surveyed articles by theme showed a variation from 40 to 200 and the number of citations of those articles showed a wider variation from 3 to 216. Selected articles from the three themes were analyzed and the first seven of the first sub-period and the first three of the second sub-period with more citations were read in full to make a synopsis of each article. Overall, the findings show that the majority of article types were models, namely mathematical, although with different sub-types for each theme. Almost all articles suggest further studies, with some mentioning it for their own author(s), which widen the diversity of the previous research. Identified research gaps concern the use of surveys to know which are the models more used by firms, the reasons for not using the models with more performance and accuracy, and which are the satisfaction levels with the outcomes of the inventories management and its effect on the improvement of the firm’s overall performance. The review ends with the limitations and contributions of the study.

Keywords: inventory management, safety stock, spare parts inventory, vendor managed inventory

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Authors: Emma J. Dale, Christina D. Buesching, Kevin R. Theis, David W. Macdonald

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This study investigates the oral and intestinal microbiomes of wild-living European badgers (Meles meles) and will relate inter-individual differences to social contact networks, somatic and reproductive fitness, varying susceptibility to bovine tuberculous (bTB) and to the olfactory advertisement. Badgers are an interesting model for this research, as they have great variation in body condition, despite living in complex social networks and having access to the same resources. This variation in somatic fitness, in turn, affects breeding success, particularly in females. We postulate that microbiota have a central role to play in determining the successfulness of an individual. Our preliminary results, characterising the microbiota of individual badgers, indicate unique compositions of microbiota communities within social groups of badgers. This basal information will inform further questions related to the extent microbiota influence fitness. Hitherto, the potential role of microbiota has not been considered in determining host condition, but also other key fitness variables, namely; communication and resistance to disease. Badgers deposit their faeces in communal latrines, which play an important role in olfactory communication. Odour profiles of anal and subcaudal gland secretions are highly individual-specific and encode information about group-membership and fitness-relevant parameters, and their chemical composition is strongly dependent on symbiotic microbiota. As badgers sniff/ lick (using their Vomeronasal organ) and over-mark faecal deposits of conspecifics, these microbial communities can be expected to vary with social contact networks. However, this is particularly important in the context of bTB, where badgers are assumed to transmit bTB to cattle as well as conspecifics. Interestingly, we have found that some individuals are more susceptible to bTB than are others. As acquired immunity and thus potential susceptibility to infectious diseases are known to depend also on symbiotic microbiota in other members of the mustelids, a role of particularly oral microbiota can currently not be ruled out as a potential explanation for inter-individual differences in infection susceptibility of bTB in badgers. Tri annually badgers are caught in the context of a long-term population study that began in 1987. As all badgers receive an individual tattoo upon first capture, age, natal as well as previous and current social group-membership and other life history parameters are known for all animals. Swabs (subcaudal ‘scent gland’, anal, genital, nose, mouth and ear) and fecal samples will be taken from all individuals, stored at -80oC until processing. Microbial samples will be processed and identified at Wayne State University’s Theis (Host-Microbe Interactions) Lab, using High Throughput Sequencing (16S rRNA-encoding gene amplification and sequencing). Acknowledgments: Gas-Chromatography/ Mass-spectrometry (in the context of olfactory communication) analyses will be performed through an established collaboration with Dr. Veronica Tinnesand at Telemark University, Norway.

Keywords: communication, energetics, fitness, free-ranging animals, immunology

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Authors: Izabela Kapsa

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Civic participation is an important aspect of democracy. The contemporary model of democracy is based on citizens' participation in political decision-making (deliberative democracy, participatory democracy). This participation takes many forms of activities like display of slogans and symbols, voting, social consultations, political demonstrations, membership in political parties or organizing civil disobedience. The countries of Central and Eastern Europe after 1989 are characterized by great social, economic and political diversity. Civil society is also part of the process of democratization. Civil society, funded by the rule of law, civil rights, such as freedom of speech and association and private ownership, was to play a central role in the development of liberal democracy. Among the many interpretations of concepts, defining the concept of contemporary democracy, one can assume that the terms civil society and democracy, although different in meaning, nowadays overlap. In the post-communist countries, the process of shaping and maturing societies took place in the context of a struggle with a state governed by undemocratic power. State fraud or repudiation of the institution is a representative state, which in the past was the only way to manifest and defend its identity, but after the breakthrough became one of the main obstacles to the development of civil society. In Central and Eastern Europe, there are many obstacles to the development of civil society, for example, the elimination of economic poverty, the implementation of educational campaigns, consciousness-related obstacles, the formation of social capital and the deficit of social activity. Obviously, civil society does not only entail an electoral turnout but a broader participation in the decision-making process, which is impossible without direct and participative democratic institutions. This article considers such broad forms of civic participation and their characteristics in Central and Eastern Europe. The paper is attempts to analyze the functioning of electronic forms of civic participation in Central and Eastern European states. This is not accompanied by a referendum or a referendum initiative, and other forms of political participation, such as public consultations, participative budgets, or e-Government. However, this paper will broadly present electronic administration tools, the application of which results from both legal regulations and increasingly common practice in state and city management. In the comparative analysis, the experiences of post-communist bloc countries will be summed up to indicate the challenges and possible goals for further development of this form of citizen participation in the political process. The author argues that for to function efficiently and effectively, states need to involve their citizens in the political decision-making process, especially with the use of electronic tools.

Keywords: Central and Eastern Europe, e-participation, e-government, post-communism

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Authors: Sandeep Kumar Agrawal, Aditi Bhattacharya, Janvie Manhas, Krushna Bhatt, Yatin Kholakiya, Nupur Khera, Ajoy Roychoudhury, Sudip Sen

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Autologous cultured explants of human oral mucosal epithelial cells (OMEC) are a potential therapeutic modality for limbal stem cell deficiency (LSCD). Injury or inflammation of the ocular surface in the form of burns, chemicals, Stevens Johnson syndrome, ocular cicatricial pemphigoid etc. can lead to destruction and deficiency of limbal stem cells. LSCD manifests in the form of severe ocular surface diseases (OSD) characterized by persistent and recurrent epithelial defects, conjuntivalisation and neovascularisation of the corneal surface, scarring and ultimately opacity and blindness. Most of the cases of OSD are associated with severe dry eye pertaining to diminished mucin and aqueous secretion. Mycophenolate mofetil (MMF) has been shown to upregulate the mucin expression in conjunctival goblet cells in vitro. The aim of this study was to evaluate the effects of MMF on mucin expression in primary cultures of oral mucosal epithelial cells. With institutional ethics committee approval and written informed consent, thirty oral mucosal epithelial tissue samples were obtained from patients undergoing oral surgery for non-malignant conditions. OMEC were grown on human amniotic membrane (HAM, obtained from expecting mothers undergoing elective caesarean section) scaffold for 2 weeks in growth media containing DMEM & Ham’s F12 (1:1) with 10% FBS and growth factors. In vitro dosage of MMF was standardised by MTT assay. Analysis of stem cell markers was done using RT-PCR while mucin mRNA expression was quantified using RT-PCR and q-PCR before and after treating cultured OMEC with graded concentrations of MMF for 24 hours. Protein expression was validated using immunocytochemistry. Morphological studies revealed a confluent sheet of proliferating, stratified oral mucosal epithelial cells growing over the surface of HAM scaffold. The presence of progenitor stem cell markers (p63, p75, β1-Integrin and ABCG2) and cell surface associated mucins (MUC1, MUC15 and MUC16) were elucidated by RT-PCR. The mucin mRNA expression was found to be upregulated in MMF treated primary cultures of OMEC, compared to untreated controls as quantified by q-PCR with β-actin as internal reference gene. Increased MUC1 protein expression was validated by immunocytochemistry on representative samples. Our findings conclude that OMEC have the ability to form a multi-layered confluent sheet on the surface of HAM similar to a cornea, which is important for the reconstruction of the damaged ocular surface. Cultured OMEC has stem cell properties as demonstrated by stem cell markers. MMF can be a novel enhancer of mucin production in OMEC. It has the potential to improve dry eye in patients undergoing OMEC transplantation for bilateral OSD. Further clinical trials are required to establish the role of MMF in patients undergoing OMEC transplantation.

Keywords: limbal stem cell deficiency, mycophenolate mofetil, mucin, ocular surface disease

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Authors: Iman Farasat, Howard M. Salis

Abstract:

Engineered genetic circuits reprogram cellular behavior to act as living computers with applications in detecting cancer, creating self-controlling artificial tissues, and dynamically regulating metabolic pathways. Phenemenological models are often used to simulate and design genetic circuit behavior towards a desired behavior. While such models assume that each circuit component’s function is modular and independent, even small changes in a circuit (e.g. a new promoter, a change in transcription factor expression level, or even a new media) can have significant effects on the circuit’s function. Here, we use statistical thermodynamics to account for the several factors that control transcriptional regulation in bacteria, and experimentally demonstrate the model’s accuracy across 825 measurements in several genetic contexts and hosts. We then employ our first principles model to design, experimentally construct, and characterize a family of signal amplifying genetic circuits (genetic OpAmps) that expand the dynamic range of cell sensors. To develop these models, we needed a new approach to measuring the in vivo binding free energies of transcription factors (TFs), a key ingredient of statistical thermodynamic models of gene regulation. We developed a new high-throughput assay to measure RNA polymerase and TF binding free energies, requiring the construction and characterization of only a few constructs and data analysis (Figure 1A). We experimentally verified the assay on 6 TetR-homolog repressors and a CRISPR/dCas9 guide RNA. We found that our binding free energy measurements quantitatively explains why changing TF expression levels alters circuit function. Altogether, by combining these measurements with our biophysical model of translation (the RBS Calculator) as well as other measurements (Figure 1B), our model can account for changes in TF binding sites, TF expression levels, circuit copy number, host genome size, and host growth rate (Figure 1C). Model predictions correctly accounted for how these 8 factors control a promoter’s transcription rate (Figure 1D). Using the model, we developed a design framework for engineering multi-promoter genetic circuits that greatly reduces the number of degrees of freedom (8 factors per promoter) to a single dimensionless unit. We propose the Ptashne (Pt) number to encapsulate the 8 co-dependent factors that control transcriptional regulation into a single number. Therefore, a single number controls a promoter’s output rather than these 8 co-dependent factors, and designing a genetic circuit with N promoters requires specification of only N Pt numbers. We demonstrate how to design genetic circuits in Pt number space by constructing and characterizing 15 2-repressor OpAmp circuits that act as signal amplifiers when within an optimal Pt region. We experimentally show that OpAmp circuits using different TFs and TF expression levels will only amplify the dynamic range of input signals when their corresponding Pt numbers are within the optimal region. Thus, the use of the Pt number greatly simplifies the genetic circuit design, particularly important as circuits employ more TFs to perform increasingly complex functions.

Keywords: transcription factor, synthetic biology, genetic circuit, biophysical model, binding energy measurement

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Authors: Julia Kapr, Andrea Rossi, Haribaskar Ramachandran, Marius Pollet, Ilka Egger, Selina Dangeleit, Katharina Koch, Jean Krutmann, Ellen Fritsche

Abstract:

It is widely acknowledged that animal models do not always represent human disease. Especially human brain development is difficult to model in animals due to a variety of structural and functional species-specificities. This causes significant discrepancies between predicted and apparent drug efficacies in clinical trials and their subsequent failure. Emerging alternatives based on 3D in vitro approaches, such as human brain spheres or organoids, may in the future reduce and ultimately replace animal models. Here, we present a human induced pluripotent stem cell (hiPSC)-based 3D neural in a vitro disease model for the Cockayne Syndrome B (CSB). CSB is a rare hereditary disease and is accompanied by severe neurologic defects, such as microcephaly, ataxia and intellectual disability, with currently no treatment options. Therefore, the aim of this study is to investigate the molecular and cellular defects found in neural hiPSC-derived CSB models. Understanding the underlying pathology of CSB enables the development of treatment options. The two CSB models used in this study comprise a patient-derived hiPSC line and its isogenic control as well as a CSB-deficient cell line based on a healthy hiPSC line (IMR90-4) background thereby excluding genetic background-related effects. Neurally induced and differentiated brain sphere cultures were characterized via RNA Sequencing, western blot (WB), immunocytochemistry (ICC) and multielectrode arrays (MEAs). CSB-deficiency leads to an altered gene expression of markers for autophagy, focal adhesion and neural network formation. Cell migration was significantly reduced and electrical activity was significantly increased in the disease cell lines. These data hint that the cellular pathologies is possibly underlying CSB. By induction of autophagy, the migration phenotype could be partially rescued, suggesting a crucial role of disturbed autophagy in defective neural migration of the disease lines. Altered autophagy may also lead to inefficient mitophagy. Accordingly, disease cell lines were shown to have a lower mitochondrial base activity and a higher susceptibility to mitochondrial stress induced by rotenone. Since mitochondria play an important role in neurotransmitter cycling, we suggest that defective mitochondria may lead to altered electrical activity in the disease cell lines. Failure to clear the defective mitochondria by mitophagy and thus missing initiation cues for new mitochondrial production could potentiate this problem. With our data, we aim at establishing a disease adverse outcome pathway (AOP), thereby adding to the in-depth understanding of this multi-faced disorder and subsequently contributing to alternative drug development.

Keywords: autophagy, disease modeling, in vitro, pluripotent stem cells

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