Search results for: regulation & smart contracts

Authors: Vlad Andrei Raducanu, Mariana Lucia Angelescu, Ion Cinca, Vasile Danut Cojocaru, Doina Raducanu

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By using smart digital tools, such as generative design (GD) and digital fabrication (DF), problems of high actuality concerning resources optimization (materials, energy, time) can be solved and applications or products of free-form type can be created. In the new digital technology materials are active, designed in response to a set of performance requirements, which impose a total rethinking of old material practices. The article presents the design procedure key steps of a free-form architectural object - a column type one with connections to get an adaptive 3D surface, by using the parametric design methodology and by exploiting the properties of conventional metallic materials. In parametric design the form of the created object or space is shaped by varying the parameters values and relationships between the forms are described by mathematical equations. Digital parametric design is based on specific procedures, as shape grammars, Lindenmayer - systems, cellular automata, genetic algorithms or swarm intelligence, each of these procedures having limitations which make them applicable only in certain cases. In the paper the design process stages and the shape grammar type algorithm are presented. The generative design process relies on two basic principles: the modeling principle and the generative principle. The generative method is based on a form finding process, by creating many 3D spatial forms, using an algorithm conceived in order to apply its generating logic onto different input geometry. Once the algorithm is realized, it can be applied repeatedly to generate the geometry for a number of different input surfaces. The generated configurations are then analyzed through a technical or aesthetic selection criterion and finally the optimal solution is selected. Endless range of generative capacity of codes and algorithms used in digital design offers various conceptual possibilities and optimal solutions for both technical and environmental increasing demands of building industry and architecture. Constructions or spaces generated by parametric design can be specifically tuned, in order to meet certain technical or aesthetical requirements. The proposed approach has direct applicability in sustainable architecture, offering important potential economic advantages, a flexible design (which can be changed until the end of the design process) and unique geometric models of high performance.

Keywords: parametric design, algorithmic procedures, free-form architectural object, sustainable architecture

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Authors: Catariny C. Aleman, Flavio B. Campos, Matheus A. Caliman, Everardo C. Mantovani

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The seedling production stage is a critical point in the vegetable production system. Obtaining high-quality seedlings is a prerequisite for subsequent cropping to occur well and productivity optimization is required. The water management is an important step in agriculture production. The adequate water requirement in horticulture seedlings can provide higher quality and increase field production. The practice of irrigation is indispensable and requires a duly adjusted quality irrigation system, together with a specific water management plan to meet the water demand of the crop. Irrigation management in seedling management requires a great deal of specific information, especially when it involves the use of inputs such as hydrorentering polymers and automation technologies of the data acquisition and irrigation system. The experiment was conducted in a greenhouse at the Federal University of Viçosa, Viçosa - MG. Tomato seedlings (Lycopersicon esculentum Mill) were produced in plastic trays of 128 cells, suspended at 1.25 m from the ground. The seedlings were irrigated by 4 micro sprinklers of fixed jet 360º per tray, duly isolated by sideboards, following the methodology developed for this work. During Phase 1, in January / February 2017 (duration of 24 days), the cultivation coefficient (Kc) of seedlings cultured in the presence and absence of hydrogel was evaluated by weighing lysimeter. In Phase 2, September 2017 (duration of 25 days), the seedlings were submitted to 4 irrigation managements (Kc, timer, 0.50 ETo, and 1.00 ETo), in the presence and absence of hydrogel and then evaluated in relation to quality parameters. The microclimate inside the greenhouse was monitored with the use of air temperature, relative humidity and global radiation sensors connected to a microcontroller that performed hourly calculations of reference evapotranspiration by Penman-Monteith standard method FAO56 modified for the balance of long waves according to Walker, Aldrich, Short (1983), and conducted water balance and irrigation decision making for each experimental treatment. Kc of seedlings cultured on a substrate with hydrogel (1.55) was higher than Kc on a pure substrate (1.39). The use of the hydrogel was a differential for the production of earlier tomato seedlings, with higher final height, the larger diameter of the colon, greater accumulation of a dry mass of shoot, a larger area of crown projection and greater the rate of relative growth. The handling 1.00 ETo promoted higher relative growth rate.

Keywords: automatic system; efficiency of water use; precision irrigation, micro sprinkler.

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Authors: Rosa S. Wong, Keith T. S. Tung, Frederick K. Ho, Winnie W. Y. Tso, King-wa Fu, Nirmala Rao, Patrick Ip

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As technology advances, exposures in early childhood are no longer confined to stimulations in the surrounding physical environments. Children nowadays are also subject to influences from the digital world. In particular, early access to game consoles can cause risks to child development, especially when the game is not developmentally appropriate for young children. Overstimulation is possible and could impair brain development. On the other hand, recreational parent-child activities, including outdoor activities and visits to museums, require child interaction with parents, which is beneficial for developing adaptive emotion regulation and social skills. Given the differences between these two types of exposures, this study investigated and compared the independent effects of early exposure to a game console and early play-based parent-child activities on children’s long-term psychosocial outcomes. This study used data from a subset of children (n=304, 142 male and 162 female) in the longitudinal cohort study, which studied the long-term impact of family socioeconomic status on child development. In 2012/13, we recruited a group of children at Kindergarten 3 (K3) randomly from Hong Kong local kindergartens and collected data regarding their duration of exposure to game console and recreational parent-child activities at that time. In 2018/19, we re-surveyed the parents of these children who were matriculated as Form 1 (F1) students (ages ranging from 11 to 13 years) in secondary schools and asked the parents to rate their children’s psychosocial problems in F1. Linear regressions were conducted to examine the associations between early exposures and adolescent psychosocial problems with and without adjustment for child gender and K3 family socioeconomic status. On average, K3 children spent about 42 minutes on a game console every day and had 2-3 recreational activities with their parents every week. Univariate analyses showed that more time spent on game consoles at K3 was associated with more psychosocial difficulties in F1 particularly more externalizing problems. The effect of early exposure to game console on externalizing behavior remained significant (B=0.59, 95%CI: 0.15 to 1.03, p=0.009) after adjusting for recreational parent-child activities and child gender. For recreational parent-child activities at K3, its effect on overall psychosocial difficulties became insignificant after adjusting for early exposure to game consoles and child gender. However, it was found to have significant protective effect on externalizing problems (B=-0.65, 95%CI: -1.23 to -0.07, p=0.028) even after adjusting for the confounders. Early exposure to game consoles has negative impact on children’s psychosocial health, whereas play-based parent-child activities can foster positive psychosocial outcomes. More efforts should be directed to propagate the risks and benefits of these activities and urge the parents and caregivers to replace child-alone screen time with parent-child play time in daily routine.

Keywords: early childhood, electronic device, parenting, psychosocial wellbeing

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Authors: Sijie Chen, Xiaofang Chen, Juan Wang, Yingying Zhang, Yu Hong, Wanyu Zhuang, Xinxin Huang, Ping Ou, Longsheng Huang

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Purpose: Autistic symptom improvement can be observed in children treated with acupuncture, but the mechanism is still being explored. In the present study, we used scalp acupuncture to treat autism rat model, and then their improvement in the abnormal behaviors and specific mechanisms behind were revealed by detecting animal behaviors, analyzing the RNA sequencing of the prefrontal cortex(PFC), and observing the ultrastructure of PFC neurons under the transmission electron microscope. Methods: On gestational day 12.5, Wistar rats were given valproic acid (VPA) by intraperitoneal injection, and their offspring were considered to be reliable rat models of autism. They were randomized to VPA or VPA-acupuncture group (n=8). Offspring of Wistar pregnant rats that were simultaneously injected with saline were randomly selected as the wild-type group (WT). VPA_acupuncture group rats received acupuncture intervention at 23 days of age for 4 weeks, and the other two groups followed without intervention. After the intervention, all experimental rats underwent behavioral tests. Immediately afterward, they were euthanized by cervical dislocation, and their prefrontal cortex was isolated for RNA sequencing and transmission electron microscopy. Results: The main results are as follows: 1. Animal behavioural tests: VPA group rats showed more anxiety-like behaviour and repetitive, stereotyped behaviour than WT group rats. While VPA group rats showed less spatial exploration ability, activity level, social interaction, and social novelty preference than WT group rats. It was gratifying to observe that acupuncture indeed improved these abnormal behaviors of autism rat model. 2. RNA-sequencing: The three groups of rats differed in the expression and enrichment pathways of multiple genes related to synaptic function, neural signal transduction, and circadian rhythm regulation. Our experiments indicated that acupuncture can alleviate the major symptoms of ASD by improving these neurological abnormalities. 3. Under the transmission electron microscopy, several lysosomes and mitochondrial structural abnormalities were observed in the prefrontal neurons of VPA group rats, which were manifested as atrophy of the mitochondrial membran, blurring or disappearance of the mitochondrial cristae, and even vacuolization. Moreover, the number of synapses and synaptic vesicles was relatively small. Conversely, the mitochondrial structure of rats in the WT group and VPA_acupuncture was normal, and the number of synapses and synaptic vesicles was relatively large. Conclusion: Acupuncture effectively improved the abnormal behaviors of autism rat model and the ultrastructure of the PFC neurons, which might worked by improving their abnormal synaptic function, synaptic plasticity and promoting neuronal signal transduction.

Keywords: autism spectrum disorder, acupuncture, animal behavior, RNA sequencing, transmission electron microscope

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Authors: E. Fabre, C. Vale, E. Pereira, C. M. Silva

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Introduction: Mercury is one of the most troublesome toxic metals responsible for the contamination of the aquatic systems due to its accumulation and bioamplification along the food chain. The 2030 agenda for sustainable development of United Nations promotes the improving of water quality by reducing water pollution and foments an enhance in wastewater treatment, encouraging their recycling and safe water reuse globally. Sorption processes are widely used in wastewater treatments due to their many advantages such as high efficiency and low operational costs. In these processes the target contaminant is removed from the solution by a solid sorbent. The more selective and low cost is the biosorbent the more attractive becomes the process. Agricultural wastes are especially attractive approaches for sorption. They are largely available, have no commercial value and require little or no processing. In this work, banana peels were tested for mercury removal from low concentrated solutions. In order to investigate the applicability of this solid, six water matrices were used increasing the complexity from natural waters to a real wastewater. Studies of kinetics and equilibrium were also performed using the most known models to evaluate the viability of the process In line with the concept of circular economy, this study adds value to this by-product as well as contributes to liquid waste management. Experimental: The solutions were prepared with Hg(II) initial concentration of 50 µg L-1 in natural waters, at 22 ± 1 ºC, pH 6, magnetically stirring at 650 rpm and biosorbent mass of 0.5 g L-1. NaCl was added to obtain the salt solutions, seawater was collected from the Portuguese coast and the real wastewater was kindly provided by ISQ - Instituto de Soldadura e qualidade (Welding and Quality Institute) and diluted until the same concentration of 50 µg L-1. Banana peels were previously freeze-drying, milled, sieved and the particles < 1 mm were used. Results: Banana peels removed more than 90% of Hg(II) from all the synthetic solutions studied. In these cases, the enhance in the complexity of the water type promoted a higher mercury removal. In salt waters, the biosorbent showed removals of 96%, 95% and 98 % for 3, 15 and 30 g L-1 of NaCl, respectively. The residual concentration of Hg(II) in solution achieved the level of drinking water regulation (1 µg L-1). For real matrices, the lower Hg(II) elimination (93 % for seawater and 81 % for the real wastewaters), can be explained by the competition between the Hg(II) ions and the other elements present in these solutions for the sorption sites. Regarding the equilibrium study, the experimental data are better described by the Freundlich isotherm (R ^ 2=0.991). The Elovich equation provided the best fit to the kinetic points. Conclusions: The results exhibited the great ability of the banana peels to remove mercury. The environmental realist conditions studied in this work, highlight their potential usage as biosorbents in water remediation processes.

Keywords: banana peels, mercury removal, sorption, water treatment

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Authors: Seema Dhariwal, Kiran Maan, Ruchi Baghel, Apoorva Sharma, Poonam Rana

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Introduction: Traumatic brain injury (TBI) is defined as the temporary or permanent alteration in brain function and pathology caused by an external mechanical force. It represents the leading cause of mortality and morbidity among children and youth individuals. Various models of TBI in rodents have been developed in the laboratory to mimic the scenario of injury. Blast overpressure injury is common among civilians and military personnel, followed by accidents or explosive devices. In addition to this, the lateral Controlled cortical impact (CCI) model mimics the blunt, penetrating injury. Method: In the present study, we have developed two different mild TBI models using blast and CCI injury. In the blast model, helium gas was used to create an overpressure of 130 kPa (±5) via a shock tube, and CCI injury was induced with an impact depth of 1.5mm to create diffusive and focal injury, respectively. C57BL/6J male mice (10-12 weeks) were divided into three groups: (1) control, (2) Blast treated, (3) CCI treated, and were exposed to different injury models. Serum was collected on Day1 and day7, followed by biphasic extraction using MTBE/Methanol/Water. Prepared samples were separated on Charged Surface Hybrid (CSH) C18 column and acquired on UHPLC-Q-TOF-MS using ESI probe with inhouse optimized parameters and method. MS peak list was generated using Markerview TM. Data were normalized, Pareto-scaled, and log-transformed, followed by multivariate and univariate analysis in metaboanalyst. Result and discussion: Untargeted profiling of lipids generated extensive data features, which were annotated through LIPID MAPS® based on their m/z and were further confirmed based on their fragment pattern by LipidBlast. There is the final annotation of 269 features in the positive and 182 features in the negative mode of ionization. PCA and PLS-DA score plots showed clear segregation of injury groups to controls. Among various lipids in mild blast and CCI, five lipids (Glycerophospholipids {PC 30:2, PE O-33:3, PG 28:3;O3 and PS 36:1 } and fatty acyl { FA 21:3;O2}) were significantly altered in both injury groups at Day 1 and Day 7, and also had VIP score >1. Pathway analysis by Biopan has also shown hampered synthesis of Glycerolipids and Glycerophospholipiods, which coincides with earlier reports. It could be a direct result of alteration in the Acetylcholine signaling pathway in response to TBI. Understanding the role of a specific class of lipid metabolism, regulation and transport could be beneficial to TBI research since it could provide new targets and determine the best therapeutic intervention. This study demonstrates the potential lipid biomarkers which can be used for injury severity diagnosis and identification irrespective of injury type (diffusive or focal).

Keywords: LipidBlast, lipidomic biomarker, LIPID MAPS®, TBI

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Authors: Markéta Chmelíková, David Ullrich, Iva Burešová

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The paper maps the possibilities and limits of diagnosing selected personality and performance characteristics of military leadership and psychology students in the context of coping with challenging situations. Individuals vary greatly inter-individually in their ability to effectively manage extreme situations, yet existing diagnostic tools are often criticized mainly for their low predictive power. Nowadays, every modern army focuses primarily on the systematic minimization of potential risks, including the prediction of desirable forms of behavior and the performance of military commanders. The context of military leadership is well known for its life-threatening nature. Therefore, it is crucial to research stress load in the specific context of military leadership for the purpose of possible anticipation of human failure in managing extreme situations of military leadership. The aim of the submitted pilot study, using an experiment of 24 hours duration, is to verify the possibilities of a specific combination of psychodiagnostic to predict people who possess suitable equipment for coping with increased stress load. In our pilot study, we conducted an experiment of 24 hours duration with an experimental group (N=13) in the bomb shelter and a control group (N=11) in a classroom. Both groups were represented by military leadership students (N=11) and psychology students (N=13). Both groups were equalized in terms of study type and gender. Participants were administered the following test battery of personality characteristics: Big Five Inventory 2 (BFI-2), Short Dark Triad (SD-3), Emotion Regulation Questionnaire (ERQ), Fatigue Severity Scale (FSS), and Impulsive Behavior Scale (UPPS-P). This test battery was administered only once at the beginning of the experiment. Along with this, they were administered a test battery consisting of the Test of Attention (d2) and the Bourdon test four times overall with 6 hours ranges. To better simulate an extreme situation – we tried to induce sleep deprivation - participants were required to try not to fall asleep throughout the experiment. Despite the assumption that a stay in an underground bomb shelter will manifest in impaired cognitive performance, this expectation has been significantly confirmed in only one measurement, which can be interpreted as marginal in the context of multiple testing. This finding is a fundamental insight into the issue of stress management in extreme situations, which is crucial for effective military leadership. The results suggest that a 24-hour stay in a shelter, together with sleep deprivation, does not seem to simulate sufficient stress for an individual, which would be reflected in the level of cognitive performance. In the context of these findings, it would be interesting in future to extend the diagnostic battery with physiological indicators of stress, such as: heart rate, stress score, physical stress, mental stress ect.

Keywords: bomb shelter, extreme situation, military leadership, psychodiagnostic

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Authors: Peter Burggräf, Matthias Dannapfel, Hanno Voet, Patrick-Benjamin Bök, Jérôme Uelpenich, Julian Hoppe

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The increasing digitalization of value chains can help companies to handle rising complexity in their processes and thereby reduce the steadily increasing planning and control effort in order to raise performance limits. Due to technological advances, companies face the challenge of smart value chains for the purpose of improvements in productivity, handling the increasing time and cost pressure and the need of individualized production. Therefore, companies need to ensure quick and flexible decisions to create self-optimizing processes and, consequently, to make their production more efficient. Lean production, as the most commonly used paradigm for complexity reduction, reaches its limits when it comes to variant flexible production and constantly changing market and environmental conditions. To lift performance limits, which are inbuilt in current value chains, new methods and tools must be applied. Digitalization provides the potential to derive these new methods and tools. However, companies lack the experience to harmonize different digital technologies. There is no practicable framework, which instructs the transformation of current value chains into digital pervasive value chains. Current research shows that a connection between lean production and digitalization exists. This link is based on factors such as people, technology and organization. In this paper, the introduced method for the determination of digitally pervasive value chains takes the factors people, technology and organization into account and extends existing approaches by a new dimension. It is the first systematic approach for the digital transformation of lean production and consists of four steps: The first step of ‘target definition’ describes the target situation and defines the depth of the analysis with regards to the inspection area and the level of detail. The second step of ‘analysis of the value chain’ verifies the lean-ability of processes and lies in a special focus on the integration capacity of digital technologies in order to raise the limits of lean production. Furthermore, the ‘digital evaluation process’ ensures the usefulness of digital adaptions regarding their practicability and their integrability into the existing production system. Finally, the method defines actions to be performed based on the evaluation process and in accordance with the target situation. As a result, the validation and optimization of the proposed method in a German company from the electronics industry shows that the digital transformation of current value chains based on lean production achieves a raise of their inbuilt performance limits.

Keywords: digitalization, digital transformation, Industrie 4.0, lean production, value chain

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Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

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Authors: Nikolai A. Khlebnikov, Vladimir N. Krasilnikov, Evgenii V. Polyakov, Anastasia A. Maltceva

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Carbon materials with advanced surfaces are widely used both in modern industry and in environmental protection. The physical-chemical nature of these materials is determined by the morphology of primary atomic and molecular carbon structures, which are the basis for synthesizing the following materials: zero-dimensional (fullerenes), one-dimensional (fiber, tubes), two-dimensional (graphene) carbon nanostructures, three-dimensional (multi-layer graphene, graphite, foams) with unique physical-chemical and functional properties. Experience shows that the microscopic morphological level is the basis for the creation of the next mesoscopic morphological level. The dependence of the morphology on the chemical way and process prehistory (crystallization, colloids formation, liquid crystal state and other) is the peculiarity of the last called level. These factors determine the consumer properties of carbon materials, such as specific surface area, porosity, chemical resistance in corrosive environments, catalytic and adsorption activities. Based on the developed ideology of thin precursor synthesis, the authors discuss one of the approaches of the porosity control of carbon-containing materials with a given aggregates morphology. The low-temperature thermolysis of precursors in a gas environment of a given composition is the basis of the above-mentioned idea. The processes of carbothermic precursor synthesis of two different compounds: tungsten carbide WC:nC and zinc oxide ZnO:nC containing an impurity phase in the form of free carbon were selected as subjects of the research. In the first case, the transition metal (tungsten) forming carbides was the object of the synthesis. In the second case, there was selected zinc that does not form carbides. The synthesis of both kinds of transition metals compounds was conducted by the method of precursor carbothermic synthesis from the organic solution. ZnO:nC composites were obtained by thermolysis of succinate Zn(OO(CH2)2OO), formate glycolate Zn(HCOO)(OCH2CH2O)1/2, glycerolate Zn(OCH2CHOCH2OH), and tartrate Zn(OOCCH(OH)CH(OH)COO). WC:nC composite was synthesized from ammonium paratungstate and glycerol. In all cases, carbon structures that are specific for diamond- like carbon forms appeared on the surface of WC and ZnO particles after the heat treatment. Tungsten carbide and zinc oxide were removed from the composites by selective chemical dissolution preserving the amorphous carbon phase. This work presents the results of investigating WC:nC and ZnO:nC composites and carbon nanopowders with tubular, tape, plate and onion morphologies of aggregates that are separated by chemical dissolution of WC and ZnO from the composites by the following methods: SEM, TEM, XPA, Raman spectroscopy, and BET. The connection between the carbon morphology under the conditions of synthesis and chemical nature of the precursor and the possibility of regulation of the morphology with the specific surface area up to 1700-2000 m2/g of carbon-structured materials are discussed.

Keywords: carbon morphology, composite materials, precursor synthesis, tungsten carbide, zinc oxide

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

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

Keywords: allergens, food, egg proteins, immunoassay

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Authors: Igor V. Grigoriev

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Understanding of biological processes encoded in fungi is instrumental in addressing future food, feed, and energy demands of the growing human population. Genomics is a powerful and quickly evolving tool to understand these processes. The Fungal Genomics Program of the US Department of Energy Joint Genome Institute (JGI) partners with researchers around the world to explore fungi in several large scale genomics projects, changing the fungal genomics landscape. The key trends of these changes include: (i) rapidly increasing scale of sequencing and analysis, (ii) developing approaches to go beyond culturable fungi and explore fungal ‘dark matter,’ or unculturables, and (iii) functional genomics and multi-omics data integration. Power of comparative genomics has been recently demonstrated in several JGI projects targeting mycorrhizae, plant pathogens, wood decay fungi, and sugar fermenting yeasts. The largest JGI project ‘1000 Fungal Genomes’ aims at exploring the diversity across the Fungal Tree of Life in order to better understand fungal evolution and to build a catalogue of genes, enzymes, and pathways for biotechnological applications. At this point, at least 65% of over 700 known families have one or more reference genomes sequenced, enabling metagenomics studies of microbial communities and their interactions with plants. For many of the remaining families no representative species are available from culture collections. To sequence genomes of unculturable fungi two approaches have been developed: (a) sequencing DNA from fruiting bodies of ‘macro’ and (b) single cell genomics using fungal spores. The latter has been tested using zoospores from the early diverging fungi and resulted in several near-complete genomes from underexplored branches of the Fungal Tree, including the first genomes of Zoopagomycotina. Genome sequence serves as a reference for transcriptomics studies, the first step towards functional genomics. In the JGI fungal mini-ENCODE project transcriptomes of the model fungus Neurospora crassa grown on a spectrum of carbon sources have been collected to build regulatory gene networks. Epigenomics is another tool to understand gene regulation and recently introduced single molecule sequencing platforms not only provide better genome assemblies but can also detect DNA modifications. For example, 6mC methylome was surveyed across many diverse fungi and the highest among Eukaryota levels of 6mC methylation has been reported. Finally, data production at such scale requires data integration to enable efficient data analysis. Over 700 fungal genomes and other -omes have been integrated in JGI MycoCosm portal and equipped with comparative genomics tools to enable researchers addressing a broad spectrum of biological questions and applications for bioenergy and biotechnology.

Keywords: fungal genomics, single cell genomics, DNA methylation, comparative genomics

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Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

Procedia PDF Downloads 127

Authors: Seyed Hamed Mousavi, Juha M. Hijmans, Reza Rajabi, Ron Diercks, Johannes Zwerver, Henk van der Worp

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Introduction: Running is one of the most popular sports activity among people. Improper sagittal plane ankle, knee and hip kinematics are considered to be associated with the increase of injury risk in runners. Motion assessing smart-phone applications are increasingly used to measure kinematics both in the field and laboratory setting, as they are cheaper, more portable, accessible, and easier to use relative to 3D motion analysis system. The aims of this study are 1) to compare the results of 3D gait analysis system and CE; 2) to evaluate the test-retest and intra-rater reliability of coach’s eye (CE) app for the sagittal plane hip, knee, and ankle angles in the touchdown and toe-off while running. Method: Twenty subjects participated in this study. Sixteen reflective markers and cluster markers were attached to the subject’s body. Subjects were asked to run at a self-selected speed on a treadmill. Twenty-five seconds of running were collected for analyzing kinematics of interest. To measure sagittal plane hip, knee and ankle joint angles at touchdown (TD) and toe off (TO), the mean of first ten acceptable consecutive strides was calculated for each angle. A smartphone (Samsung Note5, android) was placed on the right side of the subject so that whole body was simultaneously filmed with 3D gait system during running. All subjects repeated the task with the same running speed after a short interval of 5 minutes in between. The CE app, installed on the smartphone, was used to measure the sagittal plane hip, knee and ankle joint angles at touchdown and toe off the stance phase. Results: Intraclass correlation coefficient (ICC) was used to assess test-retest and intra-rater reliability. To analyze the agreement between 3D and 2D outcomes, the Bland and Altman plot was used. The values of ICC were for Ankle at TD (TRR=0.8,IRR=0.94), ankle at TO (TRR=0.9,IRR=0.97), knee at TD (TRR=0.78,IRR=0.98), knee at TO (TRR=0.9,IRR=0.96), hip at TD (TRR=0.75,IRR=0.97), hip at TO (TRR=0.87,IRR=0.98). The Bland and Altman plots displaying a mean difference (MD) and ±2 standard deviation of MD (2SDMD) of 3D and 2D outcomes were for Ankle at TD (MD=3.71,+2SDMD=8.19, -2SDMD=-0.77), ankle at TO (MD=-1.27, +2SDMD=6.22, -2SDMD=-8.76), knee at TD (MD=1.48, +2SDMD=8.21, -2SDMD=-5.25), knee at TO (MD=-6.63, +2SDMD=3.94, -2SDMD=-17.19), hip at TD (MD=1.51, +2SDMD=9.05, -2SDMD=-6.03), hip at TO (MD=-0.18, +2SDMD=12.22, -2SDMD=-12.59). Discussion: The ability that the measurements are accurately reproduced is valuable in the performance and clinical assessment of outcomes of joint angles. The results of this study showed that the intra-rater and test-retest reliability of CE app for all kinematics measured are excellent (ICC ≥ 0.75). The Bland and Altman plots display that there are high differences of values for ankle at TD and knee at TO. Measuring ankle at TD by 2D gait analysis depends on the plane of movement. Since ankle at TD mostly occurs in the none-sagittal plane, the measurements can be different as foot progression angle at TD increases during running. The difference in values of the knee at TD can depend on how 3D and the rater detect the TO during the stance phase of running.

Keywords: reliability, running, sagittal plane, two dimensional

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Authors: Fabrizio Ascione, Martina Borrelli, Rosa Francesca De Masi, Silvia Ruggiero, Giuseppe Peter Vanoli

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The refurbishment of public buildings is one of the key factors of energy efficiency policy of European States. Educational buildings account for the largest share of the oldest edifice with interesting potentialities for demonstrating best practice with regards to high performance and low and zero-carbon design and for becoming exemplar cases within the community. In this context, this paper discusses the critical issue of dealing the energy refurbishment of a university building in heating dominated climate of South Italy. More in detail, the importance of using validated models will be examined exhaustively by proposing an analysis on uncertainties due to modelling assumptions mainly referring to the adoption of stochastic schedules for occupant behavior and equipment or lighting usage. Indeed, today, the great part of commercial tools provides to designers a library of possible schedules with which thermal zones can be described. Very often, the users do not pay close attention to diversify thermal zones and to modify or to adapt predefined profiles, and results of designing are affected positively or negatively without any alarm about it. Data such as occupancy schedules, internal loads and the interaction between people and windows or plant systems, represent some of the largest variables during the energy modelling and to understand calibration results. This is mainly due to the adoption of discrete standardized and conventional schedules with important consequences on the prevision of the energy consumptions. The problem is surely difficult to examine and to solve. In this paper, a sensitivity analysis is presented, to understand what is the order of magnitude of error that is committed by varying the deterministic schedules used for occupation, internal load, and lighting system. This could be a typical uncertainty for a case study as the presented one where there is not a regulation system for the HVAC system thus the occupant cannot interact with it. More in detail, starting from adopted schedules, created according to questioner’ s responses and that has allowed a good calibration of energy simulation model, several different scenarios are tested. Two type of analysis are presented: the reference building is compared with these scenarios in term of percentage difference on the projected total electric energy need and natural gas request. Then the different entries of consumption are analyzed and for more interesting cases also the comparison between calibration indexes. Moreover, for the optimal refurbishment solution, the same simulations are done. The variation on the provision of energy saving and global cost reduction is evidenced. This parametric study wants to underline the effect on performance indexes evaluation of the modelling assumptions during the description of thermal zones.

Keywords: energy simulation, modelling calibration, occupant behavior, university building

Procedia PDF Downloads 141

Authors: Minghui Zhang, Sirine Fkaier, Sabri Fernana, Svetlana Kiseleva, Denis Kiselev

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The real-time identification of bacteria and fungi is difficult because they emit much weaker signals than pollen. In 2020, Plair developed Rapid-E+, which extends abilities of Rapid-E to detect smaller bioaerosols such as bacteria and fungal spores with diameters down to 0.3 µm, while keeping the similar or even better capability for measurements of large bioaerosols like pollen. Rapid-E+ enables simultaneous measurements of (1) time-resolved, polarization and angle dependent Mie scattering patterns, (2) fluorescence spectra resolved in 16 channels, and (3) fluorescence lifetime of individual particles. Moreover, (4) it provides 2D Mie scattering images which give the full information on particle morphology. The parameters of every single bioaerosol aspired into the instrument are subsequently analysed by machine learning. Firstly, pure species of microbes, e.g., Bacillus subtilis (a species of bacteria), and Penicillium chrysogenum (a species of fungal spores), were aerosolized in a bioaerosol chamber for Rapid-E+ training. Afterwards, we tested microbes under different concentrations. We used several steps of data analysis to classify and identify microbes. All single particles were analysed by the parameters of light scattering and fluorescence in the following steps. (1) They were treated with a smart filter block to get rid of non-microbes. (2) By classification algorithm, we verified the filtered particles were microbes based on the calibration data. (3) The probability threshold (defined by the user) step provides the probability of being microbes ranging from 0 to 100%. We demonstrate how Rapid-E+ identified simultaneously microbes based on the results of Bacillus subtilis (bacteria) and Penicillium chrysogenum (fungal spores). By using machine learning, Rapid-E+ achieved identification precision of 99% against the background. The further classification suggests the precision of 87% and 89% for Bacillus subtilis and Penicillium chrysogenum, respectively. The developed algorithm was subsequently used to evaluate the performance of microbe classification and quantification in real-time. The bacteria and fungi were aerosolized again in the chamber with different concentrations. Rapid-E+ can classify different types of microbes and then quantify them in real-time. Rapid-E+ enables classifying different types of microbes and quantifying them in real-time. Rapid-E+ can identify pollen down to species with similar or even better performance than the previous version (Rapid-E). Therefore, Rapid-E+ is an all-in-one instrument which classifies and quantifies not only pollen, but also bacteria and fungi. Based on the machine learning platform, the user can further develop proprietary algorithms for specific microbes (e.g., virus aerosols) and other aerosols (e.g., combustion-related particles that contain polycyclic aromatic hydrocarbons).

Keywords: bioaerosols, laser-induced fluorescence, Mie-scattering, microorganisms

Procedia PDF Downloads 90

Authors: Irsa Wolfram, Boruch Lorenz

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Innovative solutions with additive manufacturing applying material extrusion for functional parts necessitate innovative filaments with persistent quality. Uniform homogeneity and a consistent dispersion of particles embedded in filaments generally require multiple cycles of extrusion or well-prepared primal matter by injection molding, kneader machines, or mixing equipment. These technologies commit to dedicated equipment that is rarely at the disposal in production laboratories unfamiliar with research in polymer materials. This stands in contrast to laboratories that investigate complex material topics and technology science to leverage the potential of 3-D printing. Consequently, scientific studies in labs are often constrained to compositions and concentrations of fillersofferedfrom the market. Therefore, we introduce a prototypal laboratory methodology scalable to tailoredprimal matter for extruding ceramic composite filaments with fused filament fabrication (FFF) technology. - A desktop single-screw extruder serves as a core device for the experiments. Custom-made filaments encapsulate the ceramic fillers and serve with polylactide (PLA), which is a thermoplastic polyester, as primal matter and is processed in the melting area of the extruder, preserving the defined concentration of the fillers. Validated results demonstrate that this approach enables continuously produced and uniform composite filaments with consistent homogeneity. Itis 3-D printable with controllable dimensions, which is a prerequisite for any scalable application. Additionally, digital microscopy confirms the steady dispersion of the ceramic particles in the composite filament. - This permits a 2D reconstruction of the planar distribution of the embedded ceramic particles in the PLA matrices. The innovation of the introduced method lies in the smart simplicity of preparing the composite primal matter. It circumvents the inconvenience of numerous extrusion operations and expensive laboratory equipment. Nevertheless, it deliversconsistent filaments of controlled, predictable, and reproducible filler concentration, which is the prerequisite for any industrial application. The introduced prototypal laboratory methodology seems capable for other polymer matrices and suitable to further utilitarian particle types beyond and above ceramic fillers. This inaugurates a roadmap for supplementary laboratory development of peculiar composite filaments, providing value for industries and societies. This low-threshold entry of sophisticated preparation of composite filaments - enabling businesses to create their own dedicated filaments - will support the mutual efforts for establishing 3D printing to new functional devices.

Keywords: additive manufacturing, ceramic composites, complex filament, industrial application

Procedia PDF Downloads 106

Authors: Yasmine Salehy, Yann Leroy, Francois Cluzel, Hong-Minh Hoang, Laurence Fournaison, Anthony Delahaye, Bernard Yannou

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Taking into consideration all the life cycle of a product is now an important step in the eco-design of a product or a technology. Life cycle assessment (LCA) is a standard tool to evaluate the environmental impacts of a system or a process. Despite the improvement in refrigerant regulation through protocols, the environmental damage of refrigeration systems remains important and needs to be improved. In this paper, the environmental impacts of refrigeration systems in a typical supermarket are compared using the LCA methodology under different conditions. The system is used to provide cold at two levels of temperature: medium and low temperature during a life period of 15 years. The most commonly used architectures of supermarket cold production systems are investigated: centralized direct expansion systems and indirect systems using a secondary loop to transport the cold. The variation of power needed during seasonal changes and during the daily opening/closure periods of the supermarket are considered. R134a as the primary refrigerant fluid and two types of secondary fluids are considered. The composition of each system and the leakage rate of the refrigerant through its life cycle are taken from the literature and industrial data. Twelve scenarios are examined. They are based on the variation of three parameters, 1. location: France (Paris), Spain (Toledo) and Sweden (Stockholm), 2. different sources of electric consumption: photovoltaic panels and low voltage electric network and 3. architecture: direct and indirect refrigeration systems. OpenLCA, SimaPro softwares, and different impact assessment methods were compared; CML method is used to evaluate the midpoint environmental indicators. This study highlights the significant contribution of electric consumption in environmental damages compared to the impacts of refrigerant leakage. The secondary loop allows lowering the refrigerant amount in the primary loop which results in a decrease in the climate change indicators compared to the centralized direct systems. However, an exhaustive cost evaluation (CAPEX and OPEX) of both systems shows more important costs related to the indirect systems. A significant difference between the countries has been noticed, mostly due to the difference in electric production. In Spain, using photovoltaic panels helps to reduce efficiently the environmental impacts and the related costs. This scenario is the best alternative compared to the other scenarios. Sweden is a country with less environmental impacts. For both France and Sweden, the use of photovoltaic panels does not bring a significant difference, due to a less sunlight exposition than in Spain. Alternative solutions exist to reduce the impact of refrigerating systems, and a brief introduction is presented.

Keywords: eco-design, industrial engineering, LCA, refrigeration system

Procedia PDF Downloads 189

Authors: Monika Trzaskowska, Dorota Zielinska, Anna Lepecka, Katarzyna Neffe-Skocinska, Beata Bilska, Marzena Tomaszewska, Danuta Kolozyn-Krajewska

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Microbiological quality and food safety are important food characteristics. Regulation (EU) No 1169/2011 of the European Parliament and of the Council on the provision of food information to consumers introduces the obligation to provide information on the 'use-by' date or the date of minimum durability (DMD). The second term is the date until which the properly stored or transported foodstuff retains its physical, chemical, microbiological and organoleptic properties. The date should be preceded by 'best before'. It is used for durable products, e.g., pasta. In relation to reducing food losses, the question may be asked whether products with the date of minimum durability currently declared retain quality and safety beyond this. The aim of the study was to assess the sensory quality and microbiological safety of selected cereal products, i.e., pasta and millet after DMD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the number of bacteria from the Enterobacteriaceae family and the number of yeast and mold (TYMC) on the last day of DMD and after 1 and 3 months of storage. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of DMD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of 14 differentiators and overall quality were defined and determined. In the tested samples of millet and pasta, no pathogenic bacteria Salmonella and Listeria monocytogenes were found. The value of the distinguishing features of selected quality and microbiological safety indicators on the last DMD day was in the range of about 3-1 log cfu/g. This demonstrates the good microbiological quality of the tested food. Comparing the products, a higher number of microorganisms was found in the samples of millet. After 3 months of storage, TVC decreased in millet, while in pasta, it was found to increase in value. In both products, the number of bacteria from the Enterobacretiaceae family decreased. In contrast, the number of TYMCs increased in samples of millet, and in pasta decreased. The intensity of sensory characteristic in the studied period varied. It remained at a similar level or increased. Millet was found to increase the intensity and flavor of 'cooked porridge' 3 months after DMD. Similarly, in the pasta, the smell and taste of 'cooked pasta' was more intense. To sum up, the researched products on the last day of the minimum durability date were characterized by very good microbiological and sensory quality, which was maintained for 3 months after this date. Based on these results, the date of minimum durability of tested products could be extended. The publication was financed on the basis of an agreement with the National Center for Research and Development No. Gospostrateg 1/385753/1/NCBR/2018 for the implementation and financing of the project under the strategic research and development program 'social and economic development of Poland in the conditions of globalizing markets – GOSPOSTRATEG - acronym PROM'.

Keywords: date of minimum durability, food losses, food quality and safety, millet, pasta

Procedia PDF Downloads 161

Authors: Ilana Singer, Anastasia Lučić, Julie Leclerc

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Introduction: Tics, characterized by repetitive, sudden, non-voluntary motor movements or vocalizations, are prevalent in chronic tic disorder (CT) and Tourette Syndrome (TS). These neurodevelopmental disorders often coexist with various psychiatric conditions, leading to challenges and reduced quality of life. While medication in conjunction with behavioral interventions, such as Habit Reversal Training (HRT), Exposure Response Prevention (ERP), and Comprehensive Behavioral Intervention for Tics (CBIT), has shown efficacy, a significant proportion of patients experience persistent tics. Thus, innovative treatment approaches are necessary to improve therapeutic outcomes, such as mindfulness-based approaches. Nonetheless, the effectiveness of mindfulness-based interventions in the context of CT and TS remains understudied. Objective: The objective of this scoping review is to provide an overview of the current state of research on mindfulness-based interventions for CT and TS, identify knowledge and evidence gaps, discuss the effectiveness of mindfulness-based interventions with other treatment options, and discuss implications for clinical practice and policy development. Method: Using guidelines from Peters (2020) and the PRISMA-ScR, a scoping review was conducted. Multiple electronic databases were searched from inception until June 2023, including MEDLINE, EMBASE, PsychInfo, Global Health, PubMed, Web of Science, and Érudit. Inclusion criteria were applied to select relevant studies, and data extraction was independently performed by two reviewers. Results: Five papers were included in the study. Firstly, we found that mindfulness interventions were found to be effective in reducing anxiety and depression while enhancing overall well-being in individuals with tics. Furthermore, the review highlighted the potential role of mindfulness in enhancing functional connectivity within the Default Mode Network (DMN) as a compensatory function in TS patients. This suggests that mindfulness interventions may complement and support traditional therapeutic approaches, particularly HRT, by positively influencing brain networks associated with tic regulation and control. Conclusion: This scoping review contributes to the understanding of the effectiveness of mindfulness-based interventions in managing CT and TS. By identifying research gaps, this review can guide future investigations and interventions to improve outcomes for individuals with CT or TS. Overall, these findings emphasize the potential benefits of incorporating mindfulness-based interventions as a smaller subset within comprehensive treatment strategies. However, it is essential to acknowledge the limitations of this scoping review, such as the exclusion of a pre-established protocol and the limited number of studies available for inclusion. Further research and clinical exploration are necessary to better understand the specific mechanisms and optimal integration of mindfulness-based interventions with existing behavioral interventions for this population.

Keywords: scoping reviews, Tourette Syndrome, tics, mindfulness-based, therapy, intervention

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Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: John G. Skelly, Peter K. Dearden, Thomas W. R. Harrop, Sarah N. Inwood, Joseph Guhlin

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The Argentine stem weevil (ASW; L. bonariensis) is an economically important pasture pest in New Zealand, which causes about $200 million of damage per annum. Microctonus hyperodae (Mh), a parasite of the ASW in its natural range in South America, was introduced into New Zealand to curb the pasture damage caused by the ASW. Mh is an endoparasitic wasp that lays its eggs in the ASW halting its reproduction. Mh was initially successful at preventing ASW proliferation and reducing pasture damage. The effectiveness of Mh has since declined due to decreased parasitism rates and has resulted in increased pasture damage. Although the mechanism through which ASW has developed resistance to Mh has not been discovered, it has been proposed to be due to the different reproductive modes used by Mh and the ASW in New Zealand. The ASW reproduces sexually, whereas Mh reproduces asexually, which has been hypothesised to have allowed the ASW to ‘out evolve’ Mh. Other species within the Microctonus genus reproduce both sexually and asexually. Strains of Microctonus aethiopoides (Ma), a species closely related to Mh, reproduce either by sexual or asexual reproduction. Comparing the genomes of sexual and asexual Microctonus may allow for the identification of the mechanism of asexual reproduction and other characteristics that may improve Mh as a biocontrol agent. The genomes of Mh and three strains of Ma, two of which reproduce sexually and one reproduces asexually, have been sequenced and annotated. The French (MaFR) and Moroccan (MaMO) reproduce sexually, whereas the Irish strain (MaIR) reproduces asexually. Like Mh, The Ma strains are also used as biocontrol agents, but for different weevil species. The genomes of Mh and MaIR were subsequently upgraded using Hi-C, resulting in a set of high quality, highly contiguous genomes. A subset of the genes involved in mitosis and meiosis, which have been identified though the use of Hidden Markov Models generated from genes involved in these processes in other Hymenoptera, have been catalogued in Mh and the strains of Ma. Meiosis and mitosis genes were broadly conserved in both sexual and asexual Microctonus species. This implies that either the asexual species have retained a subset of the molecular components required for sexual reproduction or that the molecular mechanisms of mitosis and meiosis are different or differently regulated in Microctonus to other insect species in which these mechanisms are more broadly characterised. Bioinformatic analysis of the chemoreceptor compliment in Microctonus has revealed some variation in the number of olfactory receptors, which may be related to host preference. Phylogenetic analysis of olfactory receptors highlights variation, which may be able to explain different host range preferences in the Microctonus. Hi-C clustering implies that Mh has 12 chromosomes, and MaIR has 8. Hence there may be variation in gene regulation between species. Genome alignment of Mh and MaIR implies that there may be large scale genome structural variation. Greater insight into the genetics of these agriculturally important group of parasitic wasps may be beneficial in restoring or maintaining their biocontrol efficacy.

Keywords: argentine stem weevil, asexual, genomics, Microctonus hyperodae

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Authors: Yusuf Ahmad Sani, Adamu Alhaji Yakubu, Alhaji Abdullahi Jamilu, Joel Omeke, Ibrahim Jumare Sambo

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With the rising global threat of food security, cancer, and related diseases (carcinogenic) because of increased usage of inorganic substances in agricultural food production, the Ministry of Food Agriculture and Livestock of the Republic of Turkey organized an International Workshop on Organic Agriculture between 8 – 12th December 2014 at the International Agricultural Research and Training Center, Izmir. About 21 countries, including Nigeria, were invited to attend the training workshop. Several topics on organic agriculture were presented by renowned scholars, ranging from regulation, certification, crop, animal, seed production, pest and disease management, soil composting, and marketing of organic agricultural products, among others. This paper purposely selected two countries (Macedonia and Iran) out of the 21 countries to assess their level of development in terms of organic agriculture as compared to Nigeria. Macedonia, with a population of only 2.1 million people as of 2014, started organic agriculture in 2005 with only 266ha of land and has grown significantly to over 5,000ha in 2010, covering such crops as cereals (62%), forage (20%) fruit orchard (7%), vineyards (5%), vegetables (4%), oil seed and industrial crops (1%) each. Others are organic beekeeping from 110 hives to over 15,000 certified colonies. As part of government commitment, the level of government subsidy for organic products was 30% compared to the direct support for conventional agricultural products. About 19 by-laws were introduced on organic agricultural production that was fully consistent with European Union regulations. The republic of Iran, on the other hand, embarked on organic agriculture for the fact, that the country recorded the highest rate of cancer disease in the world, with over 30,000 people dying every year and 297 people diagnosed every day. However, the host country, Turkey, is well advanced in organic agricultural production and now being the largest exporter of organic products to Europe and other parts of the globe. A technical trip to one of the villages that are under the government scheme on organic agriculture reveals that organic agriculture was based on market-demand-driven and the support of the government was very visible, linking the farmers with private companies that provide inputs to them while the companies purchase the products at harvest with high premium price. However, in Nigeria, research on organic agriculture was very recent, and there was very scanty information on organic agriculture due to poor documentation and very low awareness, even among the elites. The paper, therefore, recommends that the government should provide funds to NARIs to conduct research on organic agriculture and to establish clear government policy and good pre-conditions for sustainable organic agricultural production in the country.

Keywords: organic agriculture, food security, food safety, food nutrition

Procedia PDF Downloads 50

Authors: Ekatereine Abashidze, Nino Nazarashvili, Dali Gaganidze, Oleg Gorgadze, Mariam Aznarashvili, Eter Gvritishvili

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Potato is one of the leading agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. Potato production is equal to 25,000 ha and its average yield is 20-25 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (Globodera pallida (Stone) Behrens and Globodera rostochiensis (Wollenveber) Behrens) are harmful around the world. PCN is among the most difficult plant pests to control. Cysts protected by a durable wall can survive for over 30 years . Control of PCN (G. pallida and G. rostochiensis) is regulated by Council Directive 2007/33/EE C. There was no legislative regulation of these pests in Georgia before 2016. By Resolution #302 from July 1, 2016, developed within the action plan of the DCFTA (Deep and Comprehensive Free Trade Area) the Government of Georgia established control over potato cyst nematodes. The Agreement about the legal acts approximation to EU legislation concerns the approval of rules of PCN control and research of these pests. Taking into consideration the above mentioned, it is necessary to study PCN (G. pallida and G. rostochiensis) in the potato-growing areas of Georgia. The aim of this research is to conduct survey of potato cyst nematodes (Globodera rostochiensis and G. pallida) in two geographically distinct regions of Georgia - Samtskhe - Javakheti and Svanetii and to identify the species G. Rostochiensis and G. Pallida by the morphological - morphometric and molecular methods. Soil samples were taken in each village, in a zig-zag pattern on the potato fields of the private sector, using the Metlitsky method. Samples were taken also from infested potato plant roots. To extract nematode cysts from soil samples Fanwick can be used according to standard methods by EPPO. Cysts were measured under a stereoscopic microscope (Leica M50). Identification of the nematod species was carried out according to morphological and morphometric characteristics of the cysts and larvae using appropriate protocols EPPO. For molecular identification, a multiplex PCR test was performed by the universal ITS5 and cyst nematodes’ (G. pallida, G. rostochiensis) specific primers. To identify the species of potato cyst nematodes (PCN) in two regions (Samtskhe-Javakheti and Svaneti) were taken 200 samples, among them: 80 samples in Samtskhe-Javakheti region and 120 in Svaneti region. Cysts of Globiodera spp. were revealed in 50 samples obtained from Samtskhe-Javakheti and 80 samples from Svaneti regions. Morphological, morphometric and molecular analysis of two forms of PCN found in investigated regions of Georgia shows that one form of PCN belongs to G. rostoshiensi; the second form is the different species of Globodera sp.t is the subject of future research. Despite the different geographic locations, larvae and cysts of G. rostoshiensi were found in both regions. But cysts and larvae of G. pallida were not reported. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia: Project # FR17_235.

Keywords: cyst nematode, globodera rostochiensis, globodera pallida, morphologic-morphometric measurement

Procedia PDF Downloads 200

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

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

Keywords: wine, ota removal, food safety, fining

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Authors: Hantian Wu, Bo Huang, Yuan Zeng

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Divisions between wealthy and poor, private and public landscapes are propagated by the increasing economic inequality of cities. While these are the spatial reflections of larger social issues and problems, urban design can at least employ spatial techniques that promote more inclusive rather than exclusive, overlapping rather than segregated, interlinked rather than disconnected landscapes. Indeed, the type of edge or border between urban landscapes plays a critical role in the way the environment is perceived. China experiences rapid urbanization, which poses unpredictable environmental challenges. The urban green cover and water body are under changes, which highly relevant to resident wealth and happiness. However, very limited knowledge and data on their rapid changes are available. In this regard, enhancing the monitoring of urban landscape with high-frequency method, evaluating and estimating the impacts of the urban landscape changes, and understating the driving forces of urban landscape changes can be a significant contribution for urban planning and studying. High-resolution remote sensing data has been widely applied to urban management in China. The map of urban land use map for the entire China of 2018 with 10 meters resolution has been published. However, this research focuses on the large-scale and high-resolution remote sensing land use but does not precisely focus on the seasonal change of urban covers. High-resolution remote sensing data has a long-operation cycle (e.g., Landsat 8 required 16 days for the same location), which is unable to satisfy the requirement of monitoring urban-landscape changes. On the other hand, aerial-remote or unmanned aerial vehicle (UAV) sensing are limited by the aviation-regulation and cost was hardly widely applied in the mega-cities. Moreover, those data are limited by the climate and weather conditions (e.g., cloud, fog), and those problems make capturing spatial and temporal dynamics is always a challenge for the remote sensing community. Particularly, during the rainy season, no data are available even for Sentinel Satellite data with 5 days interval. Many natural events and/or human activities drive the changes of urban covers. In this case, enhancing the monitoring of urban landscape with high-frequency method, evaluating and estimating the impacts of the urban landscape changes, and understanding the mechanism of urban landscape changes can be a significant contribution for urban planning and studying. This project aims to use the high spatiotemporal fusion of remote sensing data to create short-cycle, high-resolution remote sensing data sets for exploring the high-frequently urban cover changes. This research will enhance the long-term monitoring applicability of high spatiotemporal fusion of remote sensing data for the urban landscape for optimizing the urban management of landscape border to promoting the inclusive of the urban landscape to all communities.

Keywords: urban land cover changes, remote sensing, high spatiotemporal fusion, urban management

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Authors: Soraia V. M. de Sá, Miguel A. Faria, José O. Fernandes, Sara C. Cunha

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Cereals play a vital role in fulfilling the nutritional needs of children, supplying essential nutrients crucial for their growth and development. However, concerns arise due to children's heightened vulnerability due to their unique physiology, specific dietary requirements, and relatively higher intake in relation to their body weight. This vulnerability exposes them to harmful food contaminants, particularly mycotoxins, prevalent in cereals. Because of the thermal stability of mycotoxins, conventional industrial food processing often falls short of eliminating them. Children, especially those aged 4 months to 12 years, frequently encounter mycotoxins through the consumption of specialized food products, such as instant foods, breakfast cereals, bars, cookie snacks, fruit puree, and various dairy items. A close monitoring of this demographic group's exposure to mycotoxins is essential, as toxins ingestion may weaken children’s immune systems, reduce their resistance to infectious diseases, and potentially lead to cognitive impairments. The severe toxicity of mycotoxins, some of which are classified as carcinogenic, has spurred the establishment and ongoing revision of legislative limits on mycotoxin levels in food and feed globally. While EU Commission Regulation 1881/2006 addresses well-known mycotoxins in processed cereal-based foods and infant foods, the absence of regulations specifically addressing emerging mycotoxins underscores a glaring gap in the regulatory framework, necessitating immediate attention. Emerging mycotoxins have gained mounting scrutiny in recent years due to their pervasive presence in various foodstuffs, notably cereals and cereal-based products. Alarmingly, exposure to multiple mycotoxins is hypothesized to exhibit higher toxicity than isolated effects, raising particular concerns for products primarily aimed at children. This study scrutinizes the presence of 22 mycotoxins of the diverse range of chemical classes in 148 processed cereal-based foods, including 39 breakfast cereals, 25 infant formulas, 27 snacks, 25 cereal bars, and 32 cookies commercially available in Portugal. The analytical approach employed a modified QuEChERS procedure followed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Given the paucity of information on the risk assessment of children to multiple mycotoxins in cereal and cereal-based products consumed by children of Portugal pioneers the evaluation of this critical aspect. Overall, aflatoxin B1 (AFB1) and aflatoxin G2 (AFG2) emerged as the most prevalent regulated mycotoxins, while enniatin B (ENNB) and sterigmatocystin (STG) were the most frequently detected emerging mycotoxins.

Keywords: cereal-based products, children´s nutrition, food safety, UPLC-MS/MS analysis

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Authors: Maria Jose Granero Paris, Ana Isabel Jimenez Zarco, Agustin Pablo Alvarez Herranz

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In the industrial sector collaboration with suppliers is key to the development of innovations in the field of processes. Access to resources and expertise that are not available in the business, obtaining a cost advantage, or the reduction of the time needed to carry out innovation are some of the benefits associated with the process. However, the success of this collaborative process is compromised, when from the beginning not clearly rules have been established that govern the relationship. Abundant studies developed in the field of innovation emphasize the strategic importance of the concept of “Governance”. Despite this, there have been few papers that have analyzed how the governance process of the relationship must be designed and managed to ensure the success of the collaboration process. The lack of literature in this area responds to the wide diversity of contexts where collaborative processes to innovate take place. Thus, in sectors such as the car industry there is a strong collaborative tradition between manufacturers and suppliers being part of the value chain. In this case, it is common to establish mechanisms and procedures that fix formal and clear objectives to regulate the relationship, and establishes the rights and obligations of each of the parties involved. By contrast, in other sectors, collaborative relationships to innovate are not a common way of working, particularly when their aim is the development of process improvements. It is in this case, it is when the lack of mechanisms to establish and regulate the behavior of those involved, can give rise to conflicts, and the failure of the cooperative relationship. Because of this the present paper analyzes the similarities and differences in the processes of governance in collaboration with suppliers in the European aerospace industry With these ideas in mind, we present research is twofold: Understand the importance of governance as a key element of the success of the collaboration in the development of product and process innovations, Establish the mechanisms and procedures to ensure the proper management of the processes of collaboration. Following the methodology of the case study, we analyze the way in which manufacturers and suppliers cooperate in the development of new products and processes in two industries with different levels of technological intensity and collaborative tradition: the automotive and aerospace. The identification of those elements playing a key role to establish a successful governance and relationship management and the compression of the mechanisms of regulation and control in place at the automotive sector can be use to propose solutions to some of the conflicts that currently arise in aerospace industry. The paper concludes by analyzing the strategic implications for the aerospace industry entails the adoption of some of the practices traditionally used in other industrial sectors. Finally, it is important to highlight that in this paper are presented the first results of a research project currently in progress describing a model of governance that explains the way to manage outsourced services to suppliers in the European aerospace industry, through the analysis of companies in the sector located in Germany, France and Spain.

Keywords: supplier collaboration, supplier relationship governance, innovation management, product innovation, process innovation

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Authors: P. Asiabi, M. Shahhoseini, R. Favaedi, F. Hassani, N. Nassiri, B. Movaghar, L. Karimian, P. Eftekhariyazdi

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Introduction: Polycystic Ovary Syndrome is a common gynecologic disorder. Many factors including environment, metabolism, hormones and genetics are involved in etiopathogenesis of PCOS. Of genes that have altered expression in human reproductive system disorders are HOX family genes which act as transcription factors in regulation of cell proliferation, differentiation, adhesion and migration. Since recent evidences consider epigenetic factors as causative mechanisms of PCOS, evaluation of association between known epigenetic marks of acetylation/methylation of histone 3 (H3K9ac/me) with regulatory regions of these genes can represent better insight about PCOS. In the current study, cumulus cells (CCs) which have critical roles during folliculogenesis, oocyte maturation, ovulation and fertilization were aimed to monitor epigenetic alterations of HOX genes. Material and methods: CCs were collected from 20 PCOS patients and 20 fertile women (18-36 year) with male infertility problems referred to the Royan Institute to have ICSI under GnRH antagonist protocol. Informed consents were obtained from the participants. Thirty six hours after hCG injection, ovaries were punctured and cumulus oocyte complexes were dissected. Soluble chromatin were extracted from CCs and Chromatin Immune precipitation (ChIP) coupled with Real Time PCR was performed to quantify the epigenetic marks of histone H3K9 acetylation/methylation (H3K9ac/me) on regulatory regions of 15 members of HOX genes from A-D subfamily. Results: Obtained data showed significant increase of H3K9ac epigenetic mark on regulatory regions of HOXA1, HOXB2, HOXC4, HOXD1, HOXD3 and HOXD4 (P < 0.01) and HOXC5 (P < 0.05) and also significant decrease of H3K9ac into regulatory regions of HOXA2, HOXA4, HOXA5, HOXB1 and HOXB5 (P < 0.01) and HOXB3 (P<0.05) in PCOS patients vs. control group. On the other side, there was a significant decrease in incorporation of H3K9me level on regulatory region of HOXA2, HOXA3, HOXA4, HOXA5, HOXB3 and HOXC4 (P≤0.01) and HOXB5 (P < 0.05) in PCOS patients vs. control group. This epigenetic mark (H3K9me2) has significant increase on regulatory region of HOXB1, HOXB2, HOXC5, HOXD1, HOXD3 and HOXD4 (P ≤ 0.01) and HOXB4 (P < 0.05) in patients vs. control group. There were no significant changes in acetylation/methylation levels of H3K9 on regulatory regions of the other studied genes. Conclusion: Current study suggests that epigenetic alterations of HOX genes can be correlated with PCOS and consequently female infertility. This finding might offer additional definitions of PCOS, and eventually provides insight for novel treatments with epidrugs for this disease.

Keywords: epigenetic, HOX genes, PCOS, female infertility

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Authors: Danny Barash

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Riboswitches are RNA genetic control elements that were originally discovered in bacteria and provide a unique mechanism of gene regulation. They work without the participation of proteins and are believed to represent ancient regulatory systems in the evolutionary timescale. One of the biggest challenges in riboswitch research is that many are found in prokaryotes but only a small percentage of known riboswitches have been found in certain eukaryotic organisms. The few examples of eukaryotic riboswitches were identified using sequence-based bioinformatics search methods that include some slight structural considerations. These pattern-matching methods were the first ones to be applied for the purpose of riboswitch detection and they can also be programmed very efficiently using a data structure called affix arrays, making them suitable for genome-wide searches of riboswitch patterns. However, they are limited by their ability to detect harder to find riboswitches that deviate from the known patterns. Several methods have been developed since then to tackle this problem. The most commonly used by practitioners is Infernal that relies on Hidden Markov Models (HMMs) and Covariance Models (CMs). Profile Hidden Markov Models were also carried out in the pHMM Riboswitch Scanner web application, independently from Infernal. Other computational approaches that have been developed include RMDetect by the use of 3D structural modules and RNAbor that utilizes Boltzmann probability of structural neighbors. We have tried to incorporate more sophisticated secondary structure considerations based on RNA folding prediction using several strategies. The first idea was to utilize window-based methods in conjunction with folding predictions by energy minimization. The moving window approach is heavily geared towards secondary structure consideration relative to sequence that is treated as a constraint. However, the method cannot be used genome-wide due to its high cost because each folding prediction by energy minimization in the moving window is computationally expensive, enabling to scan only at the vicinity of genes of interest. The second idea was to remedy the inefficiency of the previous approach by constructing a pipeline that consists of inverse RNA folding considering RNA secondary structure, followed by a BLAST search that is sequence-based and highly efficient. This approach, which relies on inverse RNA folding in general and our own in-house fragment-based inverse RNA folding program called RNAfbinv in particular, shows capability to find attractive candidates that are missed by Infernal and other standard methods being used for riboswitch detection. We demonstrate attractive candidates found by both the moving-window approach and the inverse RNA folding approach performed together with BLAST. We conclude that structure-based methods like the two strategies outlined above hold considerable promise in detecting riboswitches and other conserved RNAs of functional importance in a variety of organisms.

Keywords: riboswitches, RNA folding prediction, RNA structure, structure-based methods

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