Search results for: Arvin Najafi

Authors: Ali Siami, Masoud Asayesh, Asghar Najafi, Amirhosein Hamedanian

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There are so many noise sources in power generation units that these sources can produce high-level sound noise. Therefore, sound noise reduction methods can assist these industries, especially in these days that laws related to environmental issues become more strict. In a typical power plant, so many machines and devices with high-level sound noise are arranged beside of each others. Therefore, the sound source identification and reducing the noise level can be very vital. In this paper, the procedure for designing, manufacturing and testing of a silencer-muffler used for a power plant steam vent is mentioned. This unit is placed near the residential area and so it is very important to reduce the noise emission. For this purpose, in the first step, measurements have done to identify the sound source and the frequency content of noise. The overall level of noise was so high and it was more than 120dB. Then, the appropriate noise control device is designed according to the measurement results and operational conditions. In the next step, the designed silencer-muffler has been manufactured and installed on the steam discharge of the ejector. For validation of the silencer-muffler effect, the acoustic test was done again in operating mode. Finally, the measurement results before and after the installation are compared. The results have confirmed a considerable reduction in noise level resultant of using silencer-muffler in the designed frequency range.

Keywords: silencer-muffler, sound noise control, sound measurement, steam ejector

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Authors: M. A. Akhavan-Behabadi, M. Najafi, A. Abbasi

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An empirical investigation was performed in order to study the heat transfer characteristics of R600a flow boiling inside horizontal flattened tubes and the simultaneous effect of nanoparticles on boiling heat transfer in flattened channel. Round copper tubes of 8.7 mm I.D. were deformed into flattened shapes with different inside heights of 6.9, 5.5, and 3.4 mm as test areas. The effect of different parameters such as mass flux, vapor quality and inside height on heat transfer coefficient was studied. Flattening the tube caused a significant enhancement in heat transfer performance, so that the maximum augmentation ratio of 163% was obtained in flattened channel with lowest internal height. A new correlation was developed based on the present experimental data to predict the heat transfer coefficient in flattened tubes. This correlation estimated 90% of the entire database within ±20%. The best flat channel with the point of view of heat transfer performance was selected to study the effect of nanoparticle on heat transfer enhancement. Four homogenized mixtures containing 1% weight fraction of R600a/oil with different CuO nanoparticles concentration including 0.5%, 1% and 1.5% mass fraction of R600a/oil/CuO were studied. Observations show that heat transfer was improved by adding nanoparticles, which lead to maximum enhancement of 79% compare to the pure refrigerant at the same test condition.

Keywords: nano fluids, heat transfer, flattend tube, transport phenomena

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Authors: Seyedvahid Najafi, Viliam Makis

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In recent years, maintenance optimization has attracted special attention due to the growth of industrial systems complexity. Maintenance costs are high for many systems, and preventive maintenance is effective when it increases operations' reliability and safety at a reduced cost. The novelty of this research is to consider general repair in the modeling of multi-unit series systems and solve the maintenance problem for such systems using the semi-Markov decision process (SMDP) framework. We propose an opportunistic maintenance policy for a series system composed of two main units. Unit 1, which is more expensive than unit 2, is subjected to condition monitoring, and its deterioration is modeled using a gamma process. Unit 1 hazard rate is estimated by the proportional hazards model (PHM), and two hazard rate control limits are considered as the thresholds of maintenance interventions for unit 1. Maintenance is performed on unit 2, considering an age control limit. The objective is to find the optimal control limits and minimize the long-run expected average cost per unit time. The proposed algorithm is applied to a numerical example to compare the effectiveness of the proposed policy (policy Ⅰ) with policy Ⅱ, which is similar to policy Ⅰ, but instead of general repair, replacement is performed. Results show that policy Ⅰ leads to lower average cost compared with policy Ⅱ. 

Keywords: condition-based maintenance, proportional hazards model, semi-Markov decision process, two-unit series systems

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Authors: Najafi S., Heidai R., Jamei R., Tofigh F.

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In order to study the effects of magnetic field on the root system and growth of Phaseolus vulgaris, an experiment was conducted in 2012. The possible involvement of magnetic field (MF) pretreatment in physiological factors of Phaseolus vulgaris was investigated. Seeds were subjected to 10 days with 1.8 mT of magnetic field for 1h per day. MF pretreatment decreased the plant height, fresh and dry weight, length of root and length of shoot, Chlorophyll a, Chlorophyll b and carotenoid in 10 days old seedling. In addition, activity of enzymes such as Catalase and Guaiacol peroxidase was decreased due to MF exposure. Also, the total Protein and DPPH content of the treated by magnetic field was not significantly changed in compare to control groups, while the flavonoid, Phenol and prolin content of the treated of the treated by magnetic field was significantly changed in compare to control groups. Lateral branches of roots and secondary roots increased with MF. The results suggest that pretreatment of this MF plays important roles in changes in crop productivity. In all cases there was observed a slight stimulating effect of the factors examined. The growth dynamics were weakened. The plants were shorter. Moreover, the effect of a magnetic field on the crop of Phaseolus vulgaris and its structure was small.

Keywords: carotenoid, Chlorophyll a, Chlorophyll b, DPPH, enzymes, flavonoid, germination, growth, phenol, proline, protein, magnetic field, phaseolus vulgaris

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Authors: Farzad Tofigh, Saeideh Najafi, Reza Heidari, Rashid Jamei

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In order to study the effects of magnetic field on the root system and growth of Phaseolus vulgaris, an experiment was conducted in 2012. The possible involvement of magnetic field (MF) pretreatment in physiological factors of Phaseolus vulgaris was investigated. Seeds were subjected to 10 days with 1.8 mT of magnetic field for 1h per day. MF pretreatment decreased the plant height, fresh and dry weight, length of root and length of shoot, Chlorophyll a, Chlorophyll b and carotenoid in 10 days old seedling. In addition, activity of enzymes such as Catalase and Guaiacol peroxidase was decreased due to MF exposure. Also, the total Protein and DPPH content of the treated by magnetic field was not significantly changed in compare to control groups, while the flavonoid, Phenol and prolin content of the treated of the treated by magnetic field was significantly changed in compare to control groups. Lateral branches of roots and secondary roots increased with MF. The results suggest that pretreatment of this MF plays important roles in changes in crop productivity. In all cases there was observed a slight stimulating effect of the factors examined. The growth dynamics were weakened. The plants were shorter. Moreover, the effect of a magnetic field on the crop of Phaseolus vulgaris and its structure was small.

Keywords: carotenoid, chlorophyll a, chlorophyll b, DPPH, enzymes, flavonoid, germination, growth, phenol, proline, protein, magnetic field

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Authors: Rohollah Ghasemi, Arvin Ghorbani

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This study investigates the effect of austempering temperature on microstructure and scratch behavior of the austempered heat-treated compacted graphite irons. The as-cast was used as base material for heat treatment practices. The samples were extracted from as-cast ferritic CGI pieces and were heat treated under austenitising temperature of 900°C for 60 minutes which followed by quenching in salt-bath at different austempering temperatures of 275°C, 325°C and 375°C. For all heat treatments, an austempering holding time of 30 minutes was selected for this study. Light optical microscope (LOM) and scanning electron microscope (SEM) and electron back scattered diffraction (EBSD) analysis confirmed the ausferritic matrix formed in all heat-treated samples. Microscratches were performed under the load of 200, 600 and 1000 mN using a sphero-conical diamond indenter with a tip radius of 50 μm and induced cone angle 90° at a speed of 10 μm/s at room temperature ~25°C. An instrumented nanoindentation machine was used for performing nanoindentation hardness measurement and microscratch testing. Hardness measurements and scratch resistance showed a significant increase in Brinell, Vickers, and nanoindentation hardness values as well as microscratch resistance of the heat-treated samples compared to the as-cast ferritic sample. The increase in hardness and improvement in microscratch resistance are associated with the formation of the ausferrite matrix consisted of carbon-saturated retained austenite and acicular ferrite in austempered matrix. The maximum hardness was observed for samples austempered at 275°C which resulted in the formation of very fine acicular ferrite. In addition, nanohardness values showed a quite significant variation in the matrix due to the presence of acicular ferrite and carbon-saturated retained austenite. It was also observed that the increase of austempering temperature resulted in increase of volume of the carbon-saturated retained austenite and decrease of hardness values.

Keywords: austempered CGI, austempering, scratch testing, scratch plastic deformation, scratch hardness

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Authors: Neda Najafi, Mehran Allalhesabi

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At present, the natural, geographical, physical contexts of the rural textures, which play a crucial role in making the concept behind their body, are not considered in the new designs. Despite the fundamental differences in contexts, this issue has caused that, the new rural textures in our country become similar to each other and the cohesive structure of many villages in the development of rural areas are exposed to deterioration. The villages of northern Iran are not immune from this situation and nothing have remained from their physical characteristic, and the new sections of rural areas are designed heterogeneously and regardless to the concepts behind the region's architecture, which destroys the originality of the environment. The purpose of this study is to extract the concepts and criteria that differentiate the body of the village and reveal its similarity with the same structures. In this way, understanding the underlying values is extremely useful and is considered very important to approach the new model. In the first part, the subject matters of the research (context, village and rural architecture) are defined and then the characteristics of context-oriented rural architecture and criteria that can be examined from the perspective of contextualism approach are presented. In the second part, by selecting 3 samples from the houses of Baliran village, these concepts and criteria have been evaluated in the houses of the village. The results of this study show that the characteristics of contextual rural architecture have the ability to adapt to the body of the village and can be the best model to achieve contextual architecture in this area. Therefore, by using these concepts and criteria, it is possible to achieve a type of architecture that is located along with the past architecture and, with the help of the modern facilities and environmental potentials, creates a logical and correct flow in the physical development of the rural textures.

Keywords: context, village, rural architecture, concepts and criteria of physical contextualism

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Authors: Marzieh Fotovvat, Farzaneh Najafi, Ramazan Ali Khavari-Nejad, Daryush Talei, Farhad Rejali

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Salvia leriifolia Benth. is one of the valuable and perennial medicinal plants of the Lamiaceae family, geographically growing in the south and tropical regions of Khorassan and Semnan provinces in Iran. In recent years, several medicinal properties such as antimicrobial, antifungal, anti-diabetic, analgesic, and anti-inflammatory effects have been reported from this plant. The use of elicitors such as chitosan and Arbuscular mycorrhizal fungi (AMF) symbiosis are the main methods for increasing the production of secondary metabolites, growth, and physiological factors in plants. The main aim of this study was to investigate the effects of foliar spraying applications by chitosan and/or the contribution of AMF (Glomus interaradices) on some growth factors and chlorophyll content of S. leriifolia under glasshouse conditions. The sterilized seeds were germinated by placing them into a cocopeat. After one month, seedlings that were in the 2-4 leaf stage were transferred to plastic pots (garden soil and pumice at 2:1) with or without mycorrhizal fungi. Chitosan (0, 50, 100, 200, and 400 mg L-1) was sprayed four times in the fourth month of the vegetative period. The results showed that fresh leaf weight, fresh root weight, root height, and chlorophyll content could change in the plant treated with chitosan and AMF symbiosis. So that the highest chlorophyll content and fresh weight of roots and leaves were observed in the interaction of chitosan and G. interaradices. In general, by optimizing the chitosan concentration and the use of appropriate AMF symbiosis, it is possible to improve the growth and quality of the medicinal plant S. leriifolia.

Keywords: chitosan, chlorophyll, growth factors, mycorrhiza

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Authors: S. Shafiee Ardestani, A. Najafi, N. Valizadeh, E. Payani, H. Karimian

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Objectives: Treatment of mild traumatic brain injury in emergency ward patients with any type of traumatic intracranial hemorrhage is flexible. The aim of this study is to assess the clinical outcomes of mild traumatic brain injury patients who had acute traumatic intracranial hemorrhage on initial emergency ward neuroimaging. Materials-Methods: From March 2011 to November 2012 in a retrospective cohort study we enrolled emergency ward patients with mild traumatic brain injury with Glasgow Coma Scale (GCS) scores of 14 or 15 and who had stable vital signs. Patients who had any type of intracranial hemorrhage on first head CT and repeat head CT within 24 hours were included. Patients with initial GCS < 14, injury > 24 hours old, pregnancy, concomitant non-minor injuries, and coagulopathy were excluded. Primary endpoints were neurosurgical procedures and/or death and for discharged patients, return to the emergency ward during one week. Results: Among 755 patients who were referred to the emergency ward and underwent two head CTs during first 24 hours, 302 (40%) were included. The median interval between CT scans was 6 hours (ranging 4 to 8 hours). Consequently, 135 (45%) patients had subarachnoid hemorrhage, 124 (41%) patients had subdural hemorrhage, 15 (5%) patients had epidural hemorrhage, 28 (9%) patients had cerebral contusions, and 54 (18%) patients had intra-parenchymal hemorrhage. Six of 302 patients died within 15 days of injury. 200 patients (66%) have been discharged from the emergency ward, 25 (12%) of whom returned to the emergency ward after one week. Conclusion: Discharge of the head trauma patients after a repeat head CT and brief period of observation in the emergency ward lead to early discharge of mild traumatic brain injury patients with traumatic ICH without adverse events.

Keywords: clinical outcomes, emergency ward, mild traumatic intracranial hemorrhage, Glasgow Coma Scale (GCS)

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Authors: Alireza Jalalvand, Maryam Saleh, Somayeh Behjat Khatouni, Zahra Bahri Najafi, Foroozan Fatahinia, Narges Ismailzadeh, Behrokh Farahmand

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Object: In the last two decades, the recent outbreak of Coronavirus (SARS-CoV-2) imposed a global pandemic in the world. Despite the increasing prevalence of the disease, there are no effective drugs to treat it. A suitable and rapid way to afford an effective drug and treat the global pandemic is a computational drug study. This study used molecular docking methods to examine the potential inhibition of over 50 antiviral drugs against three fundamental proteins of SARS-CoV-2. METHODS: Through a literature review, three important proteins (a key protease, RNA-dependent RNA polymerase (RdRp), and spike) were selected as drug targets. Three-dimensional (3D) structures of protease, spike, and RdRP proteins were obtained from the Protein Data Bank. Protein had minimal energy. Over 50 antiviral drugs were considered candidates for protein inhibition and their 3D structures were obtained from drug banks. The Autodock 4.2 software was used to define the molecular docking settings and run the algorithm. RESULTS: Five drugs, including indinavir, lopinavir, saquinavir, nelfinavir, and remdesivir, exhibited the highest inhibitory potency against all three proteins based on the binding energies and drug binding positions deduced from docking and hydrogen-bonding analysis. Conclusions: According to the results, among the drugs mentioned, saquinavir and lopinavir showed the highest inhibitory potency against all three proteins compared to other drugs. It may enter laboratory phase studies as a dual-drug treatment to inhibit SARS-CoV-2.

Keywords: covid-19, drug repositioning, molecular docking, lopinavir, saquinavir

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Authors: Hyvee Gean Cabuso, Arvin Taruc, Danielle Villanueva, Channela Anais Hipolito, Jia Bianca Alfonso

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Introduction: Denture adhesive aids to provide additional retention, support and comfort for patients with loose dentures, as well as for patients who seek to achieve optimal denture adhesion. But due to its growing popularity, arising oral health issues should be considered, including its possible impact that may alter the microbiological condition of the denture. Changes as such may further resolve to denture-related oral diseases that can affect the day-to-day lives of patients. Purpose: The study aims to assess and compare the microbiological status of dentures without adhesives versus dentures when adhesives were applied. The study also intends to identify the presence of specific microorganisms, their colony concentration and their possible effects on the oral microflora. This study also aims to educate subjects by introducing an alternative denture cleaning method as well as denture and oral health care. Methodology: Edentulous subjects age 50-80 years old, both physically and medically fit, were selected to participate. Before obtaining samples for the study, the alternative cleaning method was introduced by demonstrating a step-by-step cleaning process. Samples were obtained by swabbing the intaglio surface of their upper and lower prosthesis. These swabs were placed in a thioglycollate broth, which served as a transport and enrichment medium. The swabs were then processed through bacterial culture. The colony-forming units (CFUs) were calculated on MacConkey Agar Plate (MAP) and Blood Agar Plate (BAP) in order to identify and assess the microbiological status, including species identification and microbial counting. Result: Upon evaluation and analysis of collected data, the microbiological assessment of the upper dentures with adhesives showed little to no difference compared to dentures without adhesives, but for the lower dentures, (P=0.005), which is less than α = 0.05; therefore, the researchers reject (Ho) and that there is a significant difference between the mean ranks of the lower denture without adhesive to those with, implying that there is a significant decrease in the bacterial count. Conclusion: These results findings may implicate the possibility that the addition of denture adhesives may contribute to the significant decrease of microbial colonization on the dentures.

Keywords: denture, denture adhesive, denture-related, microbiological assessment

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Authors: Fatemeh Najafi

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The major stress factor limiting crop growth and development of sesame (Sesamum indicum L.) is drought stress in arid and semiarid regions of the world. For this study the effects of water stress on some qualitative and quantitative traits in sesame germplasm was conducted in the Research Farm of Seed and Plant Improvement Institute, Karaj, in the crop year. Genotypes in a randomized complete block design with three replications in two environments (moisture stress and normal) were studied in regard of the seed weight, capsule weight, grain yield, biomass, plant height, number of capsules per plant, etc. The characteristics were evaluated based on the combined analysis. Irrigation was based on first class evaporation basin. After flowering stage drought stress was applied. The water deficit reduced growth period. Days to reach full ripening decreased so that the reduction was significant at the five percent level. Drought stress reduces yield and plant biomass. Genotypes based on combined analysis of these two traits were significant at the one percent level. Genotypes differ in terms of yield stress in terms of density plots, grain yield, days to first flowering and days to the half of the cap on the confidence level of five percent and traits of days to emergence of the first capsule and days to reach full ripening at the one percent level were significant. Other traits were not significant. The correlation of traits in circumstances of stress the number of seeds per capsule has the greatest impact on performance. The sensitivity and stress tolerance index was calculated. Based on the indicators, (Fars variety) and variety Karaj were identified as the most tolerant genotypes among the studied genotypes to drought stress. The highest sensitivity indicator of stress was related to genotype (FARS).

Keywords: sesamum, drought, stress, germplasm, resistance

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Authors: Haj Najafi Leila, Tehranizadeh Mohsen

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Dependencies between diverse factors involved in probabilistic seismic loss evaluation are recognized to be an imperative issue in acquiring accurate loss estimates. Dependencies among component damage costs could be taken into account considering two partial distinct states of independent or perfectly-dependent for component damage states; however, in our best knowledge, there is no available procedure to take account of loss dependencies in story level. This paper attempts to present a method called "modal cost superposition method" for decoupling story damage costs subjected to earthquake ground motions dealt with closed form differential equations between damage cost and engineering demand parameters which should be solved in complex system considering all stories' cost equations by the means of the introduced "substituted matrixes of mass and stiffness". Costs are treated as probabilistic variables with definite statistic factors of median and standard deviation amounts and a presumed probability distribution. To supplement the proposed procedure and also to display straightforwardness of its application, one benchmark study has been conducted. Acceptable compatibility has been proven for the estimated damage costs evaluated by the new proposed modal and also frequently used stochastic approaches for entire building; however, in story level, insufficiency of employing modification factor for incorporating occurrence probability dependencies between stories has been revealed due to discrepant amounts of dependency between damage costs of different stories. Also, more dependency contribution in occurrence probability of loss could be concluded regarding more compatibility of loss results in higher stories than the lower ones, whereas reduction in incorporation portion of cost modes provides acceptable level of accuracy and gets away from time consuming calculations including some limited number of cost modes in high mode situation.

Keywords: dependency, story-cost, cost modes, engineering demand parameter

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Authors: Saheb Yousefi, Kim T. Zebehazy, Parviz Sharifi Daramadi, Tahereh Najafi Fard, Kevin Murfitt

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Context: Children with visual impairments often face challenges in their cognitive, motor, and social development. Sensory-motor and spatial perception therapies can be beneficial for these children, but many existing programs only focus on a limited set of therapies. This study aims to investigate the effectiveness of a rehabilitation program for sensory- motor and spatial perception in reducing the fear of mobility in visually impaired children. Research Aim: The aim of this study is to determine if a rehabilitation program based on parent-child interaction can reduce the fear of mobility in visually impaired children. Methodology: This study uses a semi-experimental approach with an uneven control group design. Visually impaired children aged 10 to 14 and their parents from the Board of the Blind and Visually Impaired in Tehran Province were included in the study. The sample was divided into experimental and control groups, with a total of 30 participants. The experimental group participated in a rehabilitation program for sensory-motor and spatial perception based on parent-child interaction, while the control group did not receive this intervention. Data was collected using questionnaires on transportation issues and analyzed using multivariate and univariate mixed analysis of variance tests. Findings: The analysis of the data showed that the fear of movement was significantly improved in the experimental group compared to the control group after the intervention. Theoretical Importance: This study highlights the effectiveness of a rehabilitation program for sensory- motor and spatial perception based on parent-child interaction in reducing the fear of mobility in visually impaired children. It contributes to the existing knowledge by demonstrating the positive impact of this type of intervention on the cognitive, motor, and social development of these children. Data Collection and Analysis Procedures: Data was collected through the use of questionnaires administered to the children before and after the intervention. The data was analyzed using multivariate and univariate mixed analysis of variance tests to examine the effects of the rehabilitation program. Questions Addressed: This study addresses the question of whether a rehabilitation program based on parent-child interaction can reduce the fear of mobility in visually impaired children. Conclusion: The findings of this study support the effectiveness of the sensory-motor and spatial perception rehabilitation program based on parent-child interaction in reducing the fear of movement in visually impaired children. This intervention can be considered as a suitable method to enhance the fear of mobility in these children.

Keywords: vision impairment, sensory-motor rehabilitation, space perception, parent-child interaction, fear of movement.

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Authors: Shabnam Najafi, Metin Turkay

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Network design problem (NDP) is used to determine the set of optimal values for certain pre-specified decision variables such as capacity expansion of nodes and links by optimizing various system performance measures including safety, congestion, and accessibility. The designed transportation network should improve objective functions defined for the system by considering the route choice behaviors of network users at the same time. The NDP studies mostly investigated the random demand and route selection constraints separately due to computational challenges. In this work, we consider both random demand and route selection constraints simultaneously. This work presents a nonlinear stochastic model for land use and road network design problem to address the development of different functional zones in urban areas by considering both cost function and air pollution. This model minimizes cost function and air pollution simultaneously with random demand and stochastic route selection constraint that aims to optimize network performance via road capacity expansion. The Bureau of Public Roads (BPR) link impedance function is used to determine the travel time function in each link. We consider a city with origin and destination nodes which can be residential or employment or both. There are set of existing paths between origin-destination (O-D) pairs. Case of increasing employed population is analyzed to determine amount of roads and origin zones simultaneously. Minimizing travel and expansion cost of routes and origin zones in one side and minimizing CO emission in the other side is considered in this analysis at the same time. In this work demand between O-D pairs is random and also the network flow pattern is subject to stochastic user equilibrium, specifically logit route choice model. Considering both demand and route choice, random is more applicable to design urban network programs. Epsilon-constraint is one of the methods to solve both linear and nonlinear multi-objective problems. In this work epsilon-constraint method is used to solve the problem. The problem was solved by keeping first objective (cost function) as the objective function of the problem and second objective as a constraint that should be less than an epsilon, where epsilon is an upper bound of the emission function. The value of epsilon should change from the worst to the best value of the emission function to generate the family of solutions representing Pareto set. A numerical example with 2 origin zones and 2 destination zones and 7 links is solved by GAMS and the set of Pareto points is obtained. There are 15 efficient solutions. According to these solutions as cost function value increases, emission function value decreases and vice versa.

Keywords: epsilon-constraint, multi-objective, network design, stochastic

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Authors: S. Najafi, E. Bertini, M. Pezzotti, G.B. Tornielli, S. Zenoni

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Grapevine is an important agricultural fruit crop plant consumed worldwide and with a key role in the global economy. Grapevine is strongly affected by both biotic and abiotic stresses, which impact grape growth at different stages, such as during plant and berry development and pre- and post-harvest, consequently causing significant economic losses. Recently global warming has propelled the anticipation of the onset of berry ripening, determining the reduction of a grape color and increased volatilization of aroma compounds. Climate change could negatively alter the physiological characteristics of the grape and affect the berry and wine quality. Modern plant breeding can provide tools such as genome editing for improving grape resilience traits while maintaining intact the viticultural and oenological quality characteristics of the genotype. This study aims at developing a platform for genome editing application in grapevine plants with the final goal to improve berry quality, biotic, and abiotic resilience traits. We chose to directly deliver ribonucleoproteins (RNP, preassembled Cas protein and guide RNA) into plant protoplasts, and, from these cell structures, regenerate grapevine plants edited in specific selected genes controlling traits of interest. Edited plants regenerated by somatic embryogenesis from protoplasts will then be sequenced and molecularly characterized. Embryogenic calli of Sultana and Shiraz cultivars were initiated from unopened leaves of in-vitro shoot tip cultures and from stamens, respectively. Leaves were placed on NB2 medium while stamens on callus initiation medium (PIV) medium and incubated in the dark at 28 °C for three months. Viable protoplasts, tested by FDA staining, isolated from embryogenic calli were cultured by disc method at 1*105 protoplasts/ml. Mature well-shaped somatic embryos developed directly in the protoplast culture medium two months later and were transferred in the light into to shooting medium for further growth. Regenerated plants were then transferred to the greenhouse; no phenotypic alterations were observed when compared to non in-vitro cultured plants. The performed experiments allowed to established an efficient protocol of embryogenic calli production, protoplast isolation, and regeneration of the whole plant through somatic embryogenesis in both Sultana and Shiraz. Regenerated plants, through direct somatic embryogenesis deriving from a single cell, avoid the risk of chimerism during the regeneration process, therefore improving the genome editing process. As pre-requisite of genome editing, an efficient method for transfection of protoplast by yellow fluorescent protein (YFP) marker genes was also established and experiments of direct delivery of CRISPR–Cas9 ribonucleoproteins (RNPs) in protoplasts to achieve efficient DNA-free targeted mutations are in progress.

Keywords: CRISPR-cas9, plant regeneration, protoplast isolation, Vitis vinifera

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