Search results for: defense enzymes

Authors: Shahram Nanekarania, Majid Goodarzia

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Calpastatin and callipyge have been known as one of the candidate genes in meat quality and quantity. Calpastatin gene has been located to chromosome 5 of sheep and callipyge gene has been localized in the telomeric region on ovine chromosome 18. The objective of this study was identification of calpastatin and callipyge genes polymorphism and analysis of genotype structure in population of Lori sheep kept in Iran. Blood samples were taken from 120 Lori sheep breed and genomic DNA was extracted by salting out method. Polymorphism was identified using the PCR-RFLP technique. The PCR products were digested with MspI and FaqI restriction enzymes for calpastatin gene and callipyge gene, respectively. In this population, three patterns were observed and AA, AB, BB genotype have been identified with the 0.32, 0.63, 0.05 frequencies for calpastatin gene. The results obtained for the callipyge gene revealed that only the wild-type allele A was observed, indicating that only genotype AA was present in the population under consideration.

Keywords: polymorphism, calpastatin, callipyge, PCR-RFLP, Lori sheep

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Authors: Serap Pektas

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Ataxia telangiectasia mutated (ATM) is a serine-threonine kinase, which is the major regulator of the DNA damage response. ATM is activated upon the formation of DNA double-strand breaks (DSBs) in the cells. ATM phosphorylates the proteins involved in apoptotic responses, cell cycle checkpoint control, DNA repair, etc. Tumor protein p53, known as p53 is one of these proteins that phosphorylated by ATM. Phosphorylation of p53 at Ser15 residue leads to p53 stabilization in the cells. Often enzymes activity is affected by hydrogen ion concentration (pH). In order to find the optimal pH range for ATM activity, steady-state kinetic assays were performed at acidic and basic pH ranges. Ser15 phosphorylation of p53 is determined by using ELISA. The results indicated that the phosphorylation rate was better at basic pH range compared with the acidic pH range. This could be due to enzyme stability, or enzyme-substrate interaction is pH dependent.

Keywords: ataxia telangiectasia mutated, DNA double strand breaks, DNA repair, tumor protein p53

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Authors: Sibgha Noreen, Muhammad Salim Akhter, Seema Mahmood

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Salt stress is an abiotic stress that causes a heavy toll on growth and development and also reduces the productivity of arable and horticultural crops. Globally, a quarter of total arable land has fallen prey to this menace, and more is being encroached because of the usage of brackish water for irrigation purposes. Though barley is categorized as salt-tolerant crop, but cultivars show a wide genetic variability in response to it. In addressing salt stress, silicon nutrition would be a facile tool for enhancing salt tolerant to sustain crop production. A greenhouse study was conducted to evaluate the response of barley (Hordeum vulgare L.) cultivars to silicon nutrition under salt stress. The treatments included [(a) four barley cultivars (Jou-87, B-14002, B-14011, B-10008); (b) two salt levels (0, 200 mM, NaCl); and (c) two silicon levels (0, 200ppm, K2SiO3. nH2O), arranged in a factorial experiment in a completely randomized design with 16 treatments and repeated 4 times. Plants were harvested at 15 days after exposure to different experimental salinity and silicon foliar conditions. Results revealed that various physiological and biochemical attributes differed significantly (p<0.05) in response to different treatments and their interactive effects. Cultivar “B-10008” excelled in biological yield, chlorophyll constituents, antioxidant enzymes, and grain yield compared to other cultivars. The biological yield of shoot and root organs was reduced by 27.3 and 26.5 percent under salt stress, while it was increased by 14.5 and 18.5 percent by exogenous application of silicon over untreated check, respectively. The imposition of salt stress at 200 mM caused a reduction in total chlorophyll content, chl ‘a’ , ‘b’ and ratio a/b by 10.6,16.8,17.1 and 7.1, while spray of 200 ppm silicon improved the quantum of the constituents by 10.4,12.1,10.2,10.3 over untreated check, respectively. The quantum of free amino acids and protein content was enhanced in response to salt stress and the spray of silicon nutrients. The amounts of superoxide dismutase, catalases, peroxidases, hydrogen peroxide, and malondialdehyde contents rose to 18.1, 25.7, 28.1, 29.5, and 17.6 percent over non-saline conditions under salt stress. However, the values of these antioxidants were reduced in proportion to salt stress by 200 ppm silicon applied as rooting medium on barley crops. The salt stress caused a reduction in the number of tillers, number of grains per spike, and 100-grain weight to the amount of 29.4, 8.6, and 15.8 percent; however, these parameters were improved by 7.1, 10.3, and 9.6 percent by foliar spray of silicon over untreated crop, respectively. It is concluded that the barley cultivar “B-10008” showed greater tolerance and adaptability to saline conditions. The yield of barley crops could be potentiated by a foliar spray of 200 ppm silicon at the vegetative growth stage under salt stress.

Keywords: salt stress, silicon nutrition, chlorophyll constituents, antioxidant enzymes, barley crop

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Authors: Stephanie M. De Oliveira

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For a long time, migrations are characterized as a difficult problem to solve. Various phenomena throughout human history caused personnel migrations, whether by the free will of migrants or not. Nowadays, governments that seek to give these people protection and dignity, either to asylum or to build a new life in a different country, make refugee protection. At present, a large amount of people, have been crossing their country's borders by land, air or sea, becoming refugees and seeking a new life away from fear, threat or violence they suffered in their country of origin. It is known that some countries have already instituted rights and rules for refugees who wish to become citizens in the country to which they immigrated, even though this is not what happens in most cases. The article will be based on research made with UN Refugee Agency (UNHCR) material as well as will analyze the interaction of the Turkish government with the European Union. Since Turkey is not part of the Union, it will be understood how the interaction was made, as well as the search for consensus, and not only humanitarian but also financial aid. The treatment of refugees and the defense of human rights within the country will also be considered.

Keywords: refugees, Turkey, asylum seekers, United Nations

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Authors: Raymond Oriebe Anyasi, Harrison Atagana

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The use of a plant in the detoxification of several toxin is been known to be enhanced by various microbial endophytes which have been reported to be contained in plants growing in any contaminated soil. Plants in their natural state are mostly colonized by endophytes which in the process forms symbiotic associations with the host plants. These benefits that the endophytes offer to the plants include amongst others to: Enhance plants growth through the production of various phytohormones; increase in the resistance of environmental stresses; produce important bioactive metabolites; help in the fixing of nitrogen in the plants organelles; help in the metal translocation and accumulation in plants; assist in the production of enzymes involves the degradation of organic contaminants. Therefore recognizing these natural processes of the microbes will enable the understanding of the effective mechanism for enhanced phytoremediation. The aim of this study was to survey the progressiveness in the study involving endophyte-assisted phytoremediation of contaminants; highlighting various pollutants, the plants used, the endophytes studied as well as the type of interaction between the plants and the microbes so as to proffer a better future prospect for the technology.

Keywords: phytoremediation, endophytes, microbes, pollution, environmental management, plants

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Authors: M. R. Aziz, W. Kuntjoro, N. V. David

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This paper presents the ballistic terminal study of the non-filled and water-filled aluminum tank. The objective was to determine the failure stages for both cases. The tank was impacted by fragment simulating projectile (FSP) with 260 m/s for non-filled and 972 m/s for water-filled. The aluminum tank was 3 mm thick, 150 mm wide and 750 mm long. The ends of the tank were closed with two polymethyl methacrylate (PMMA) windows. The test was conducted at the Science and Technology Research Institute for Defense (STRIDE) Batu Arang, Selangor, Malaysia. The results showed four main stages for non-filled tank, which were first contact between FSP and the tank, partially perforated, fully perforated with FSP and plug still intact and lastly fully perforated with FSP and plug separated. Meanwhile, for the water-filled tank, there were seven main stages, which were first contact between FSP and the tank, partial perforation, full perforation, drag phase, cavity phase, bounce wave event and the collapse of the cavity.

Keywords: fragment simulating projectile, high speed camera, tensile test, terminal ballistic

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Authors: C. Fortin, D. Tokmeninova, O. Ushakova

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Among 500 Russian universities Skolkovo Institute of Science and Technology (Skoltech) is one of the youngest (established in 2011), quite small and vastly international, comprising 20 percent of international students and 70 percent of faculty with significant academic experience at top-100 universities (QS, THE). The institute has emerged from close collaboration with MIT and leading Russian universities. Skoltech is an entirely English speaking environment. Skoltech curriculum plans of ten Master programs are based on the CDIO learning outcomes model. However, despite the Institute’s unique focus on industrial innovations and startups, one of the main challenges has become an evident large proportion of nearly half of MSc graduates entering PhD programs at Skoltech or other universities rather than industry or entrepreneurship. In order to increase the share of students joining the industrial sector after graduation, Skoltech started implementing a number of unique practices with a focus on employers’ expectations incorporated into the curriculum redesign. In this sense, extended knowledge exchange with industrial partners via collaboration in learning activities, industrial projects and assessments became essential for students’ headway into industrial and entrepreneurship pathways. Current academic curriculum includes the following types of components based on extended knowledge exchange with industrial partners: innovation workshop, industrial immersion, special industrial tracks, MSc defenses. Innovation workshop is a 4 week full time diving into the Skoltech vibrant ecosystem designed to foster innovators, focuses on teamwork, group projects, and sparks entrepreneurial instincts from the very first days of study. From 2019 the number of mentors from industry and startups significantly increased to guide students across these sectors’ demands. Industrial immersion is an exclusive part of Skoltech curriculum where students after the first year of study spend 8 weeks in an industrial company carrying out an individual or team project and are guided jointly by both Skoltech and company supervisors. The aim of the industrial immersion is to familiarize students with relevant needs of Russian industry and to prepare graduates for job placement. During the immersion a company plays the role of a challenge provider for students. Skoltech has started a special industrial track comprising deep collaboration with IPG Photonics – a leading R&D company and manufacturer of high-performance fiber lasers and amplifiers for diverse applications. The track is aimed to train a new cohort of engineers and includes a variety of activities for students within the “Photonics” MSc program. It is expected to be a successful story and used as an example for similar initiatives with other Russian high-tech companies. One of the pathways of extended knowledge exchange with industrial partners is an active involvement of potential employers in MSc Defense Committees to review and assess MSc thesis projects and to participate in defense procedures. The paper will evaluate the effect and results of the above undertaken measures.

Keywords: Curriculum redesign, knowledge exchange model, learning outcomes framework, stakeholder engagement

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Authors: Aliyu Adamu, Fahrul Huyop, Roswanira Abdul Wahab, Mohd Shahir Shamsir

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Halogenated organic compounds occur in huge amount in biosphere. This is attributable to the diverse use of halogen-based compounds in the synthesis of various industrially important products. Halogenated compound is toxic and may persist in the environment, thereby causing serious health and environmental pollution problems. L-2-haloacid dehalogenases (EC 3.8.1.2) catalyse the specific cleavage of carbon-halogen bond in L-isomers of halogenated compounds, which consequently reverse the effects of environmental halogen-associated pollution. To enhance the efficiency and utility of these enzymes, this study investigates the catalytic amino acid residues and the molecular functional mechanism of DehL, by classical molecular dynamic simulations, MM-PBSA and ab initio fragments molecular orbital (FMO) calculations. The results of the study will serve as the basis for the molecular engineering of the enzyme.

Keywords: DehL, Functional mechanism, Catalytic residues, L-2-haloacid dehalogenase

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Authors: Fadwa Chtioui

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Vital Pulp Therapy (VPT) is nowadays challenging the deep-rooted dogma of root canal treatment, being the only therapeutic option for permanent teeth diagnosed with irreversible pulpitis or carious pulp exposure. Histologic and clinical research has shown that compromised dental pulp can be treated without the full removal or excavation of all healthy pulp, and the outcome of the partial or full pulpotomy followed by a Tricalcium-Silicate-based dressing seems to show promising results in maintaining pulp vitality and preserving affected teeth in the long term. By reviewing recent advances in the techniques of VPT and their clinical effectiveness and safety in permanent teeth with irreversible Pulpitis, this work provides a new understanding of pulp pathophysiology and defense mechanisms and will reform dental practitioners' decision-making in treating irreversible pulpits from root canal therapy to vital pulp therapy by taking advantage of the biological effects of Tricalcium Silicate materials.

Keywords: irreversible pulpitis, vital pulp therapy, pulpotomy, Tricalcium Silicate

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Authors: Ahmad Manbohi, Seyyed Hamid Ahmadi

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A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R^² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.

Keywords: biological fluids, biomarkers, microfluidic paper-based electrochemical biosensors, Multiplex

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Authors: Isil Kerti, Onur Okur, Sibel Daglilar, Recep Calin

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Particulate reinforced aluminium matrix composites have gained more importance in automotive, aeronautical and defense industries due to their specific properties like as low density, high strength and stiffness, good fatigue strength, dimensional stability at high temperature and acceptable tribological properties. In this study, 2014 Aluminium alloy used as a matrix material and B₄C and SiC were selected as reinforcements components. For production of composites materials, vacuum infiltration method was used. In the experimental studies, the reinforcement volume ratios were defined by mixing as totally 10% B₄C and SiC. Aging treatment (T6) was applied to the specimens. The effect of T6 treatment on hardness was determined by using Brinell hardness test method. The effects of the aging treatment on microstructure and chemical structure were analysed by making XRD, SEM and EDS analysis on the specimens.

Keywords: metal matrix composite, vacumm infiltration method, aluminum metal matrix, mechanical feature

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Authors: Haowei Chen, Kaiqi Xiong

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This paper aims to answer how malware will propagate in Wireless Mesh Networks (WMNs) and how communication radius and distributed density of nodes affects the process of spreading. The above analysis is essential for devising network-wide strategies to counter malware. We answer these questions by developing an improved dynamical system that models malware propagation in the area where nodes were uniformly distributed. The proposed model captures both the spatial and temporal dynamics regarding the malware spreading process. Equilibrium and stability are also discussed based on the threshold of the system. If the threshold is less than one, the infected nodes disappear, and if the threshold is greater than one, the infected nodes asymptotically stabilize at the endemic equilibrium. Numerical simulations are investigated about communication radius and distributed density of nodes in WMNs, which allows us to draw various insights that can be used to guide security defense.

Keywords: Bluetooth security, malware propagation, wireless mesh networks, stability analysis

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Authors: Anthony George Armiger II

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This paper focuses on the counternarcotic strategies of US government agencies in Afghanistan from 2001-2014. Despite a heavy US presence in the country, Afghanistan currently accounts for 80% of opium production worldwide and remains a key contributor to the global drug market. This paper argues that the divergent counternarcotic strategies of various US government agencies on the ground in Afghanistan are a product of the organizational differences amongst those agencies and that those differences can challenge the implementation of counternarcotics policies in Afghanistan. To gain a more in-depth perspective, this paper analyzes the counternarcotic strategies of two US government agencies in Afghanistan; the United States Department of Defense (DoD) and the Drug Enforcement Administration (DEA). Utilizing the framework of the organizational behavior model of organizational theory, this paper will highlight the varying organizational interests, opinions, standard operating procedures, and routines of both of the government agencies. The paper concludes with implications on counternarcotics, as well as the counterinsurgency in Afghanistan and provides recommendations for future research on foreign policy and counternarcotics.

Keywords: Afghanistan, drug policy, organizational theory, United States foreign policy

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Authors: Mohamed Fadzlee Sulaiman, Mohd Zabri Adil Talib, Aswami Fadillah Mohd Ariffin

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Each individual organization has their own mechanism to build up cyber defense capability in protecting their information infrastructures from data breaches and cyber espionage. But, we can not deny the possibility of failing to detect and stop cyber attacks especially for those targeting credential information and intellectual property (IP). In this paper, we would like to share the modern approach of effective digital forensic methodology in order to identify the artifacts in tracing the trails of evidence while mitigating the infection from the target machine/s. This proposed approach will suit the digital forensic investigation to be conducted while resuming the business critical operation after mitigating the infection and minimizing the risk from the identified attack to transpire. Therefore, traditional digital forensics methodology has to be improvised to be proactive which not only focusing to discover the root caused and the threat actor but to develop the relevant mitigation plan in order to prevent from the same attack.

Keywords: digital forensic, detection, eradication, targeted attack, malware

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Authors: Iraj Musa Dawaari

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Australia and Singapore signed the Comprehensive Strategic Partnership (CSP) agreement on 29th June 2015. This agreement aims to strengthen security, defense, diplomatic, economic, education, and innovation cooperation, as well as people-to-people relations between the two states. The purpose of this study is to examine rationales underpinning the (2015 CSP) agreement in order to better understand the national and foreign policy priorities of both Australia and Singapore in the period leading up to the signing of the agreement and in the period since. This research project will seek to establish how both countries’ national and foreign policy priorities have developed in recent years in the light of growing tensions between China and the US in the Indo-Pacific region and how the CSP agreement has influenced these priorities. This project also seeks to better understand and analyse why both states signed the CSP agreement in 2015.

Keywords: Australia, Singapore, foreign policy, partnership, USA, China, Indo-Pacific Region

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Authors: Edward W. Staley, Corban G. Rivera, Ashley J. Llorens

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Advances in reinforcement learning (RL) have resulted in recent breakthroughs in the application of artificial intelligence (AI) across many different domains. An emerging landscape of development environments is making powerful RL techniques more accessible for a growing community of researchers. However, most existing frameworks do not directly address the problem of learning in complex operating environments, such as dense urban settings or defense-related scenarios, that incorporate distributed, heterogeneous teams of agents. To help enable AI research for this important class of applications, we introduce the AI Arena: a scalable framework with flexible abstractions for distributed multi-agent reinforcement learning. The AI Arena extends the OpenAI Gym interface to allow greater flexibility in learning control policies across multiple agents with heterogeneous learning strategies and localized views of the environment. To illustrate the utility of our framework, we present experimental results that demonstrate performance gains due to a distributed multi-agent learning approach over commonly-used RL techniques in several different learning environments.

Keywords: reinforcement learning, multi-agent, deep learning, artificial intelligence

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Authors: Jayson Vicente

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The young men and women of Philippine Military Academy Cadet Corps Armed Forces of the Philippines (PMA CCAFP) are bred to be front liners and last line of defense during war and times of peace; as such, they must be equipped with the most practical and most effective combat-ready Martial Arts and Combative skills to effectively fulfill their duty, as well as to protect and safeguard themselves to continue serving the people and their country. This study shall assess the current Martial Arts and Combative Program of the PMA CCAFP using descriptive methodology by interviews and floating questionnaires. The current Martial Arts and Combative Program of the PMA CCAFP with all of the subjects involved are more sports inclined rather than combat-equipped. Picking the best from each subject used in the program, this study seeks to recommend improvements or create a better Martial Arts and Combative Program that will satisfy the objective of producing Martial Arts combatant graduates. A good Martial Arts and Combative Program for PMA is essential to prepare them for what lies ahead, which is unforgiving and no rules to pacify threat.

Keywords: combative, martial arts, military, program

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Authors: Homayun Ghasemi, Mojtaba Yousefirad, Mozhgan Farzamisepehr

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Heavy metals are among soil pollutant resources that in case of accumulation in the soil and absorption by the plant, enter into the food chain and poison the plants or the people who consume those plants. This research was performed in order to examine the role of cadmium as a heavy metal in the activity of catalase and peroxidase as well as protein concentration in Trifolium resupinatum L. based on a randomized block design with three repetitions. The used treatments included consumption of Cd (NO3)2 at four levels, namely, 0, 100, 200, and 300 ppm. The plants under study were treated for 10 days. The results of the study showed that catalase activity decreased by the increase of cadmium. Moreover, peroxidase activity increased by an increase inthe consumption of cadmium. The analysis of protein level showed that plantlet protein decreased in high cadmium concentrations. The findings also demonstrated that cadmium concentration in roots was higher than in shoots.

Keywords: catalase, heavy metal, peroxidase, protein

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Authors: Azzaz Siham

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Essential oils are, by definition, secondary metabolites produced by plants as a means of defense against phytophagous pests. This work aims to study the insecticidal effect of the essential oil of three plants: Phoenician juniper Juniperus phoenicea; the Niaouli Melaleuca quinquenervia and the wild carrot Daucus carota L, on the chickpea weevil Callosobruchus maculatus F, which is known as a formidable pest of legumes. Essential oils are obtained by hydrodistillation. The study carried out in the laboratory concerning the insecticidal activity of these essential oils by contact and inhalation effect on C.maculatus gave important results, especially for the essential oil of Juniperus phoenicea for the contact test; and for the inhalation test, the essential oil of Melaleuca quinquenervia shows remarkable insecticidal activity compared to the other two oils. The results of these tests showed a very interesting action. The essential oils used very significantly describe the lifespan of adults.

Keywords: essential oils, juniperus phoenicea, melaleuca quinquenervia, daucus carota, Callosobruchus maculatus

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Authors: Asit K. Sikary, O. P. Murty

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Adipocere formation is a modification of the process of putrefaction. It consists mainly of saturated fatty acids, formed by the post-mortem hydrolysis and hydrogenation of body fats with the help of bacterial enzymes in the presence of warmth, moisture and anaerobic bacteria. In temperate climate, it takes weeks to develop while in India it starts to begin within 4-5 days. In this study, we have collected cases with adipocere formation, which were from the South Delhi region (average room temperature 27-390C) and autopsied at our centre. Details of the circumstances of the death, cause and time of death, surrounding environment and demographic profile of the deceased were taken into account. Total 16 cases were included in this study. Adipocere formation was predominantly present over cheeks, shoulder, breast, flanks, buttocks, and thighs. Out of 16, 11 cases were found in a dry atmosphere, 5 cases were brought from the water. There were 5 cases in which adipocere formation was seen in less than 2 days, and among them, in 1 case, as early as one day. This study showed that adipocere formation can be seen as early as 1 day in a hot and humid environment.

Keywords: adipocere, drowning, hanging, humid environment, strangulation, subtropical climate

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Authors: Moreno Liso Lourdes

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The industrial design protects the appearance of part or all of a product resulting from the features of, in particular, the lines, contours, colors, shape, texture or materials of the product itself or its ornamentation. The industrial property offers a different answer depending on the characteristics of the shape object of protection possible, including the trademark and industrial design. There are certain cases where the trademark right invalidate the exclusive right of the industrial design. This can occur in the following situations: 1st) collected as a sign design and trademarked; and 2nd) you want to trademark and protected as a form design (either registered or unregistered). You can either get a trade mark or design right in the same sign or form, provided it meets the legal definition of brand and design and meets the requirements imposed for the protection of each of them, even able to produce an overlap of protection. However, this double protection does not have many advantages. It is, therefore, necessary to choose the best form of legal protection according to the most adequate ratios. The diversity of rights that can use the creator of an industrial design to protect your job requires you to make a proper selection to prevent others, especially their competitors, taking advantage of the exclusivity that guarantees the law. It is necessary to choose between defending the interests of the parties through a judicial or extrajudicial procedure when the conflict arises. In this paper, we opted for the defense through mediation.

Keywords: industrial design, ADR, Law, EUIPO

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Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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Authors: I. Tulay Cagatay

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Blue-green microalgae cyanobacteria, which are important photosynthetic organisms of aquatic ecosystems, are the primary sources of many bioactive compounds such as proteins, carbohydrates, lipids, vitamins and enzymes that can be used as antimicrobial and antiviral agents. Some of these organisms are nowadays used directly in the food, cosmetic and pharmaceutical industry, or in aquaculture and biotechnological approaches like biofuel or drug therapy. Finding the effective, environmental friendly chemotropic and antimicrobial agents to control fish pathogens are crucial in a country like Turkey which has a production capacity of about 240 thousand tons of cultured fish and has 2377 production farms and which is the second biggest producer in Europe. In our study, we tested the antimicrobial activity of cyanobacterium spp. against some fish pathogens Aeromonas hydrophila and Yersinia ruckeri that are important pathogens for rainbow trout farms. Agar disk diffusion test method was used for studying antimicrobial activity on pathogens. Both tested microorganisms have shown antimicrobial activity positively as the inhibition zones were 0.45 mm and 0.40 mm respectively.

Keywords: fish pathogen, cyanobacteria, antimicrobial activity, trout

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Authors: Pauline Ribert, Gaelle Roudaut, Sebastien Dupont, Laurent Beney

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Yeasts, anhydrobiotic organisms, can survive extreme water disturbances, thanks to the prolonged and reversible suspension of their cellular activity as well as the establishment of a defense arsenal. This property is exploited by many industrialists. One of the protection systems implemented by yeast is the vitrification of its cytoplasm by trehalose. The thermal resistance of dry yeasts is a crucial parameter for their use. However, studies on the thermal resistance of dry yeasts are often based on yeasts produced in laboratory conditions with non-optimal drying processes. We, therefore, propose a study on the thermal resistance of industrial dry yeasts in relation to their thermophysical properties. Heat stress was applied at three temperatures (50, 75, and 100°C) for 10, 30, or 60-minute treatments. The survival of yeasts to these treatments was estimated, and their thermophysical properties were studied by differential scanning calorimetry. The industrial dry yeasts resisted 60 minutes at 50°C and 75°C and 10 minutes at a temperature close to 100°C. At 100°C, yeast was above their glass transition temperature. Industrial dry yeasts are therefore capable of withstanding high thermal stress if maintained in a specific thermophysical state.

Keywords: dry yeast, glass transition, thermal resistance, vitrification

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Authors: M. A. Siti Faridah, K. Muhammad, H. M. Ghazali, Y. A. Yusof

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This study was conducted to investigate the effects of three variables namely types of cell wall degrading enzymes (Viscozyme L, Pectinex Ultra SP-L, Rapidase PAC, Rohament CL and Rohapect PTE) at varying concentrations (0.25-3% v/w) and times (30 min-24 h) on the zinc (Zn-) amaranth purees. Liquefaction treatment of the Zn-amaranth purees with Viscozyme (1% v/w at pH 5 and 45ºC for 3 h) was found to be the best procedure, which produced Zn-amaranth puree with low viscosity (8.60 mPas). Zn-amaranth purees were also found to have the highest metallo-chlorophyll derivative contents (0.16 mg/g), free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH) values (12.49 mM (TE)/g fresh weight) and ferric reducing antioxidant power (FRAP) values (4.57 mM (TE)/g fresh weight) within 3 h of liquefaction. Other physicochemical properties of the enzyme-liquefied Zn-amaranth purees indicated that lightness (L*) (12.54), greenness a*/b* (-0.30), reducing sugar (103.88 mg/mL) and soluble dietary fibre (5.94%) of the purees were higher compared to that of nonenzyme-liquefied amaranth purees.

Keywords: amaranth, antioxidant, chlorophyll derivative, enzymatic liquefaction

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Authors: Han-Bin Kim, Sooim Shin, Moonsung Choi

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There is a growing interest in introducing a desired functional group on enzymes in the field of protein engineering. In here, various redox centers were newly created using histidine tag, which is widely used for protein purification, plus cobalt in one of cupredoxins, amicyanin. The coordination of Cobalt-His tag and reactivity of the Co²⁺ loaded His-tag also were characterized. 3xHis-tag, 6xHis-tag, and 9xHis-tag were introduced on amicyanin by site-directed mutagenesis, and then Co²⁺ was loaded on each His-tagged amicyanin. The spectral changes at 330 nm corresponding to cobalt binding on His-tag site indicated the binding ratio of 3xHis-tag, 6xHis-tag, and 9xHis-tag to cobalt as 1:1, 1:2, 1:3 respectively. Based on kinetic studies of binding cobalt to 3xHis-tag, 6xHis-tag, and 9xHis-tagged amicyanin, the nature of the sites was elucidated. In addition, internal electron transfer properties between Cu¹⁺ site and engineered site of amicyanin were determined. These results provide insight into improvement of metal coordination and alternation of the redox properties of metal as a new catalytic site on proteins.

Keywords: amicyanin, cobalt, histidine, protein engineering

Procedia PDF Downloads 144

Authors: K. Yu Vlasova, M. A. Abakumov, H. Wishwarsao, M. Sokolsky, N. V. Nukolova, A. G. Majouga, Y. I. Golovin, N. L. Klyachko, A. V. Kabanov

Abstract:

Introduction:Nowadays pharmaceutical medicine is aimed to create systems for combined therapy, diagnostic, drug delivery and controlled release of active molecules to target cells. Magnetic nanoparticles (MNPs) are used to achieve this aim. MNPs can be applied in molecular diagnostics, magnetic resonance imaging (T1/T2 contrast agents), drug delivery, hyperthermia and could improve therapeutic effect of drugs. The most common drug containers, containing MNPs, are liposomes, micelles and polymeric molecules bonded to the MNPs surface. Usually superparamagnetic nanoparticles are used (the general diameter is about 5-6 nm) and all effects of high frequency magnetic field (MF) application are based on Neel relaxation resulting in heating of surrounded media. In this work we try to develop a new method to improve drug release from MNPs under super low frequency MF. We suppose that under low frequency MF exposures the Brown’s relaxation dominates and MNPs rotation could occur leading to conformation changes and release of bioactive molecules immobilized on MNPs surface.The aim of this work was to synthesize different systems with active drug (biopolymers coated MNPs nanoclusters with immobilized enzymes and doxorubicin (Dox) loaded magnetic liposomes/micelles) and investigate the effect of super low frequency MF on these drug containers. Methods: We have synthesized MNPs of magnetite with magnetic core diameter 7-12 nm . The MNPs were coated with block-copolymer of polylysine and polyethylene glycol. Superoxide dismutase 1 (SOD1) was electrostatically adsorbed on the surface of the clusters. Liposomes were prepared as follow: MNPs, phosphatidylcholine and cholesterol were dispersed in chloroform, dried to get film and then dispersed in distillated water, sonicated. Dox was added to the solution, pH was adjusted to 7.4 and excess of drug was removed by centrifugation through 3 kDa filters. Results: Polylysine coated MNPs formed nanosized clusters (as observed by TEM) with intensity average diameter of 112±5 nm and zeta potential 12±3 mV. After low frequency AC MF exposure we observed change of immobilized enzyme activity and hydrodynamic size of clusters. We suppose that the biomolecules (enzymes) are released from the MNPs surface followed with additional aggregation of complexes at the MF in medium. Centrifugation of the nanosuspension after AC MF exposures resulted in increase of positive charge of clusters and change in enzyme concentration in comparison with control sample without MF, thus confirming desorption of negatively charged enzyme from the positively charged surface of MNPs. Dox loaded magnetic liposomes had average diameter of 160±8 nm and polydispersity index (PDI) 0.25±0.07. Liposomes were stable in DW and PBS at pH=7.4 at 370C during a week. After MF application (10 min of exposure, 50 Hz, 230 mT) diameter of liposomes raised to 190±10 nm and PDI was 0.38±0.05. We explain this by destroying and/or reorganization of lipid bilayer, that leads to changes in release of drug in comparison with control without MF exposure. Conclusion: A new application of low frequency AC MF for drug delivery and controlled drug release was shown. Investigation was supported by RSF-14-13-00731 grant, K1-2014-022 grant.

Keywords: magnetic nanoparticles, low frequency magnetic field, drug delivery, controlled drug release

Procedia PDF Downloads 464

Authors: Vinay Kumar K. S., Shravani V., Spoorthi G., Udith K. S., Divya T. M., Venkatesha M.

Abstract:

In this work, the mathematical modeling, design and analysis of a 2×2 microstrip patch antenna array (MSPA) antenna configuration is presented. Array utilizes a tiny strip antenna module with two vertical slots for 5G applications at an operating frequency of 5.3 GHz. The proposed array of antennas where the phased array antenna systems (PAAS) are used ubiquitously everywhere, from defense radar applications to commercial applications like 5G/6G. Microstrip patch antennae with slot arrays for linear polarisation parallel and perpendicular to the axis, respectively, are fed through transverse slots in the side wall of the circular waveguide and fed through longitudinal slots in the small wall of the rectangular waveguide. The microstrip patch antenna is developed using Ansys HFSS (High-Frequency Structure Simulator), this simulation tool. The maximum gain of 6.14 dB is achieved at 5.3 GHz for a single MSPA. For 2×2 array structure, a gain of 7.713 dB at 5.3 GHz is observed. Such antennas find many applications in 5G devices and technology.

Keywords: Ansys HFSS, gain, return loss, slot array, microstrip patch antenna, 5G antenna

Procedia PDF Downloads 88

Authors: Suchao Donpudsa, Suwattana Visetnan, Anchalee Tassanakajon, Vichien Rimphanitchayakit

Abstract:

The Single Wap Domain (SWD) is a type III crustin antimicrobial peptide whose function is to defense the host animal against the bacterial infection by means of antimicrobial and antiproteinase activities. A study of LvSWD from Litopenaeus vannamei is reported herein about its activities and function against bacteria, particularly the Vibrio parahaemolyticus AHPND (VPAHPND) that causes acute hepatopancreatic necrosis disease. The over-expressed mature recombinant (r)LvSWD exhibits antimicrobial activity against both Gram-positive and Gram-negative bacteria, especially VPAHPND. With four times the MIC of rLvSWD, the treated post larval shrimp infected by VPAHPND is able to survive longer with the 50% survival rate as long as 78 h as compared to 36 h of the infected shrimp without rLvSWD. To a certain extent, we have demonstrated that the rLvSWD can be applied to protect the post larval shrimp.

Keywords: crustin, Litopenaeus vannamei, Vibrio parahaemolyticus AHPND, antimicrobial activity

Procedia PDF Downloads 193

Authors: Li Minghui, Min Shaorong, Zhang Jun

Abstract:

This paper aims for studying the operational efficiency of surface warship’s motorized evasion to acoustic homing torpedo. It orderly developed trajectory model, self-guide detection model, vessel evasion model, as well as anti-torpedo error model in three-dimensional space to make up for the deficiency of precious researches analyzing two-dimensionally confrontational models. Then, making use of the Monte Carlo method, it carried out the simulation for the confrontation process of evasion in the environment of MATLAB. At last, it quantitatively analyzed the main factors which determine vessel’s survival probability. The results show that evasion relative bearing and speed will affect vessel’s survival probability significantly. Thus, choosing appropriate evasion relative bearing and speed according to alarming range and alarming relative bearing for torpedo, improving alarming range and positioning accuracy and reducing the response time against torpedo will improve the vessel’s survival probability significantly.

Keywords: acoustic homing torpedo, vessel evasion, monte carlo method, torpedo defense, vessel's survival probability

Procedia PDF Downloads 426