971 resultados para Quantification of Glutamate


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Plusieurs tests médicaux, comme celui du dépistage du cancer du sein, se basent sur l’observation de section tissulaire sous un microscope. Ces tests se basent sur l’interprétation d’un spécialiste et les résultats peuvent varier d’un expert à un autre dû la subjectivité des observations. L’utilisation d’une technique analytique offrant une quantification et une identification de cibles moléculaires dans une section tissulaire permettrait aux experts de produire des diagnostics plus objectifs et diminuerait possiblement le nombre de faux diagnostics. Les travaux présentés dans ce mémoire portent sur le développement d’une technique SPRi-MALDI-IMS permettant l’imagerie en deux dimensions de protéines contenues dans une section tissulaire. La MALDI-IMS est la technique de choix pour l’imagerie de biomolécules dans les sections tissulaires. Par contre, elle ne parvient pas à elle seule à quantifier de façon absolue le matériel adsorbé à la surface. Donc, le couplage de la MALDI-IMS avec la SPRi permet la quantification absolue de protéines en deux dimensions et crée une technique répondant aux besoins des experts médicaux. Pour ce faire, nous avons étudié, l’effet de la chimie de surface sur la nature et la quantité de matériel adsorbé à la surface du capteur. De plus, la cinétique de transfert des protéines du tissu vers le capteur a dû être optimisée afin de produire des empreintes correspondant au tissu d’origine, afin d’atteindre la gamme dynamique des instruments SPRi et MALDI-IMS. La technique résultante de ces optimisations permet d’obtenir les premières images quantitatives et qualitatives de protéines en deux dimensions d’une seule section tissulaire.

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Les neurones dopaminergiques (DA) de la substance noire compacte (SNc) et de l’aire tegmentaire ventrale (ATV) développent des contacts de type synaptique et non synaptique. Malgré de nombreux travaux sur la synaptogénèse en général, aucune méthode autre que la microscopie électronique, n’a été développée pour quantifier les varicosités synaptiques et asynaptiques issues des neurones DA. L’objectif principal de ce projet était de développer une méthode d’analyse et de quantification des varicosités synaptiques et asynaptiques des neurones DA. L’hypothèse proposée est qu’il devait être possible de détecter la présence de synapses en visualisant la colocalisation d’une protéine présynaptique telle que synaptotagmine 1 (SYT1) avec un marqueur post-synaptique tel que la postsynaptic density protein 95 (PSD95). Pour ce faire, nous avons préparé des cultures primaires de neurones DA à l’aide d’une lignée de souris transgéniques exprimant la protéine fluorescente verte (GFP) sous le contrôle du promoteur de la tyrosine hydroxyalse (TH). Nous avons ensuite visualisé les terminaisons axonales à l'aide de marquages immunocytochimiques de protéines pré et post-synaptiques. L’analyse quantitative des images a été effectuée avec le logiciel de traitement d’image Image-J. Nos résultats montrent que, via l’association d’un marqueur présynaptique tel que SYT1 avec un marqueur postsynaptique tel que PSD95, seule une minorité des terminaisons établies par les neurones DA sont de type synaptique. En contraste, des neurones glutamatergiques du cortex, établissent une majorité de terminaisons associées à un marqueur postsynaptique. Nos résultats valident donc la mise en place d'une technique d'analyse permettant de quantifier la proportion de terminaisons synaptiques et asynaptiques établies par les neurones DA.

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The aim of the present study was to investigate the in vitro gastric stability of colistin sulfate (CS) and its antimicrobial activity against Escherichia coli and to study the impact of ETEC O149: F4 (K88) infection in pigs on CS intestinal absorption. The stability profile of CS was evaluated in a simulated gastric fluid (SGF). Antimicrobial activity of CS and its degradation products were examined in a 96-well polystyrene microplate model. The effect of experimental infection with ETEC O149: F4 on CS intestinal absorption was determined by quantification of CS systemic concentration using a validated LC–MS/MS method. A rapid degradation of CS accompanied by an increase in CS antimicrobial activity by comparison with non-degraded CS (P < 0.0001) was observed in SGF. Additionally, CS levels were not quantifiable in systemic circulation using a highly sensitive method and concurrent oral challenge did not affect CS absorption in an induction model of subclinical post-weaning diarrhea (PWD).

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Les histones sont des protéines nucléaires hautement conservées chez les cellules des eucaryotes. Elles permettent d’organiser et de compacter l’ADN sous la forme de nucléosomes, ceux-ci representant les sous unités de base de la chromatine. Les histones peuvent être modifiées par de nombreuses modifications post-traductionnelles (PTMs) telles que l’acétylation, la méthylation et la phosphorylation. Ces modifications jouent un rôle essentiel dans la réplication de l’ADN, la transcription et l’assemblage de la chromatine. L’abondance de ces modifications peut varier de facon significative lors du developpement des maladies incluant plusieurs types de cancer. Par exemple, la perte totale de la triméthylation sur H4K20 ainsi que l’acétylation sur H4K16 sont des marqueurs tumoraux spécifiques a certains types de cancer chez l’humain. Par conséquent, l’étude de ces modifications et des événements determinant la dynamique des leurs changements d’abondance sont des atouts importants pour mieux comprendre les fonctions cellulaires et moléculaires lors du développement de la maladie. De manière générale, les modifications des histones sont étudiées par des approches biochimiques telles que les immuno-buvardage de type Western ou les méthodes d’immunoprécipitation de la chromatine (ChIP). Cependant, ces approches présentent plusieurs inconvénients telles que le manque de spécificité ou la disponibilité des anticorps, leur coût ou encore la difficulté de les produire et de les valider. Au cours des dernières décennies, la spectrométrie de masse (MS) s’est avérée être une méthode performante pour la caractérisation et la quantification des modifications d’histones. La MS offre de nombreux avantages par rapport aux techniques traditionnelles. Entre autre, elle permet d’effectuer des analyses reproductibles, spécifiques et facilite l’etude d’un large spectre de PTMs en une seule analyse. Dans cette thèse, nous présenterons le développement et l’application de nouveaux outils analytiques pour l’identification et à la quantification des PTMs modifiant les histones. Dans un premier temps, une méthode a été développée pour mesurer les changements d’acétylation spécifiques à certains sites des histones. Cette méthode combine l’analyse des histones intactes et les méthodes de séquençage peptidique afin de déterminer les changements d’acétylation suite à la réaction in vitro par l’histone acétyltransférase (HAT) de levure Rtt109 en présence de ses chaperonnes (Asf1 ou Vps75). Dans un second temps, nous avons développé une méthode d’analyse des peptides isomériques des histones. Cette méthode combine la LC-MS/MS à haute résolution et un nouvel outil informatique appelé Iso-PeptidAce qui permet de déconvoluer les spectres mixtes de peptides isomériques. Nous avons évalué Iso-PeptidAce avec un mélange de peptides synthétiques isomériques. Nous avons également validé les performances de cette approche avec des histones isolées de cellules humaines érythroleucémiques (K562) traitées avec des inhibiteurs d’histones désacétylases (HDACi) utilisés en clinique, et des histones de Saccharomyces cerevisiae liées au facteur d’assemblage de la chromatine (CAF-1) purifiées par chromatographie d’affinité. Enfin, en utilisant la méthode présentée précédemment, nous avons fait une analyse approfondie de la spécificité de plusieurs HATs et HDACs chez Schizosaccharomyces pombe. Nous avons donc déterminé les niveaux d’acétylation d’histones purifiées à partir de cellules contrôles ou de souches mutantes auxquelles il manque une HAT ou HDAC. Notre analyse nous a permis de valider plusieurs cibles connues des HATs et HDACs et d’en identifier de nouvelles. Nos données ont également permis de définir le rôle des différentes HATs et HDACs dans le maintien de l’équilibre d’acétylation des histones. Dans l’ensemble, nous anticipons que les méthodes décrites dans cette thèse permettront de résoudre certains défis rencontrés dans l’étude de la chromatine. De plus, ces données apportent de nouvelles connaissances pour l’élaboration d’études génétiques et biochimiques utilisant S. pombe.

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Solid phase extraction (SPE) is a powerful technique for preconcentration/removal or separation of trace and ultra trace amounts of toxic and nutrient elements. SPE effectively simplifies the labour intensive sample preparation, increase its reliability and eliminate the clean up step by using more selective extraction procedures. The synthesis of sorbents with a simplified procedure and diminution of the risks of errors shows the interest in the areas of environmental monitoring, geochemical exploration, food, agricultural, pharmaceutical, biochemical industry and high purity metal designing, etc. There is no universal SPE method because the sample pretreatment depends strongly on the analytical demand. But there is always an increasing demand for more sensitive, selective, rapid and reliable analytical procedures. Among the various materials, chelate modified naphthalene, activated carbon and chelate functionalized highly cross linked polymers are most important. In the biological and environmental field, large numbers of samples are to be analysed within a short span of time. Hence, online flow injection methods are preferred as they allow extraction, separation, identification and quantification of many numbers of analytes. The flow injection online preconcentration flame AAS procedure developed allows the determination of as low as 0.1 µg/l of nickel in soil and cobalt in human hair samples. The developed procedure is precise and rapid and allows the analysis of 30 samples per hour with a loading time of 60 s. The online FI manifold used in the present study permits high sampling, loading rates and thus resulting in higher preconcentration/enrichment factors of -725 and 600 for cobalt and nickel respectively with a 1 min preconcentration time compared to conventional FAAS signal. These enrichment factors are far superior to hitherto developed on line preconcentration procedures for inorganics. The instrumentation adopted in the present study allows much simpler equipment and low maintenance costs compared to costlier ICP-AES or ICP-MS instruments.

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The present study aimed at critically looking at the current practice of the installation of compacted clay liner using bentonite enhanced sand (BES). The application of bentonite is currently the most accepted practice for lining purposes. The ideal bentonite sand combination, which satisfies the liner requirements is 20% bentonite and 80% sand, was selected as one of the liner materials for the investigation of development of desiccation cracks. Locally available sundried marine clay and its combination with bentonite were also included in the study. The desiccation tests on liner materials were conducted for wet/dry cycles to simulate the seasonal variations. Digital image processing techniques were used to measure the crack intensity factor (CIF), a useful and effective parameter for quantification of desiccation cracking. The repeatability of the tests could be well established, as the variation in CIF values of identical samples had a very narrow range of 0 to 2%. The studies on the development of desiccation cracks showed that the CIF of bentonite enhanced sand mixture (BES) was 18.09%, 39.75% and 21.22% for the first, second and third cycles respectively, while it was only 9.83%, 7.52% and 4.58% respectively for sun dried marine clay (SMC). Thus the locally available, alternate liner material suggested, viz SMC, is far superior to BES, when subjected to alternate wet/dry cycles. Further, the improvement of these liner materials when amended with randomly distributed fibre reinforcements was also investigated. Three types of fibres ,namely nylon fibre, polypropylene monofilament and polypropylene fibre mesh were used for the study of fibre amended BES and SMC.The influence of these amendments on the properties of the above liner materials is also studied. The results showed that there is definite improvement in the properties of the liner materials when it is reinforced with discrete random fibres. The study also proved that the desiccation cracks could be controlled with the help of fibre reinforcement.

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We present a novel approach to computing the orientation moments and rheological properties of a dilute suspension of spheroids in a simple shear flow at arbitrary Peclct number based on a generalised Langevin equation method. This method differs from the diffusion equation method which is commonly used to model similar systems in that the actual equations of motion for the orientations of the individual particles are used in the computations, instead of a solution of the diffusion equation of the system. It also differs from the method of 'Brownian dynamics simulations' in that the equations used for the simulations are deterministic differential equations even in the presence of noise, and not stochastic differential equations as in Brownian dynamics simulations. One advantage of the present approach over the Fokker-Planck equation formalism is that it employs a common strategy that can be applied across a wide range of shear and diffusion parameters. Also, since deterministic differential equations are easier to simulate than stochastic differential equations, the Langevin equation method presented in this work is more efficient and less computationally intensive than Brownian dynamics simulations.We derive the Langevin equations governing the orientations of the particles in the suspension and evolve a procedure for obtaining the equation of motion for any orientation moment. A computational technique is described for simulating the orientation moments dynamically from a set of time-averaged Langevin equations, which can be used to obtain the moments when the governing equations are harder to solve analytically. The results obtained using this method are in good agreement with those available in the literature.The above computational method is also used to investigate the effect of rotational Brownian motion on the rheology of the suspension under the action of an external force field. The force field is assumed to be either constant or periodic. In the case of con- I stant external fields earlier results in the literature are reproduced, while for the case of periodic forcing certain parametric regimes corresponding to weak Brownian diffusion are identified where the rheological parameters evolve chaotically and settle onto a low dimensional attractor. The response of the system to variations in the magnitude and orientation of the force field and strength of diffusion is also analyzed through numerical experiments. It is also demonstrated that the aperiodic behaviour exhibited by the system could not have been picked up by the diffusion equation approach as presently used in the literature.The main contributions of this work include the preparation of the basic framework for applying the Langevin method to standard flow problems, quantification of rotary Brownian effects by using the new method, the paired-moment scheme for computing the moments and its use in solving an otherwise intractable problem especially in the limit of small Brownian motion where the problem becomes singular, and a demonstration of how systems governed by a Fokker-Planck equation can be explored for possible chaotic behaviour.

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The hazards associated with major accident hazard (MAH) industries are fire, explosion and toxic gas releases. Of these, toxic gas release is the worst as it has the potential to cause extensive fatalities. Qualitative and quantitative hazard analyses are essential for the identitication and quantification of the hazards associated with chemical industries. This research work presents the results of a consequence analysis carried out to assess the damage potential of the hazardous material storages in an industrial area of central Kerala, India. A survey carried out in the major accident hazard (MAH) units in the industrial belt revealed that the major hazardous chemicals stored by the various industrial units are ammonia, chlorine, benzene, naphtha, cyclohexane, cyclohexanone and LPG. The damage potential of the above chemicals is assessed using consequence modelling. Modelling of pool fires for naphtha, cyclohexane, cyclohexanone, benzene and ammonia are carried out using TNO model. Vapor cloud explosion (VCE) modelling of LPG, cyclohexane and benzene are carried out using TNT equivalent model. Boiling liquid expanding vapor explosion (BLEVE) modelling of LPG is also carried out. Dispersion modelling of toxic chemicals like chlorine, ammonia and benzene is carried out using the ALOHA air quality model. Threat zones for different hazardous storages are estimated based on the consequence modelling. The distance covered by the threat zone was found to be maximum for chlorine release from a chlor-alkali industry located in the area. The results of consequence modelling are useful for the estimation of individual risk and societal risk in the above industrial area.Vulnerability assessment is carried out using probit functions for toxic, thermal and pressure loads. Individual and societal risks are also estimated at different locations. Mapping of threat zones due to different incident outcome cases from different MAH industries is done with the help of Are GIS.Fault Tree Analysis (FTA) is an established technique for hazard evaluation. This technique has the advantage of being both qualitative and quantitative, if the probabilities and frequencies of the basic events are known. However it is often difficult to estimate precisely the failure probability of the components due to insufficient data or vague characteristics of the basic event. It has been reported that availability of the failure probability data pertaining to local conditions is surprisingly limited in India. This thesis outlines the generation of failure probability values of the basic events that lead to the release of chlorine from the storage and filling facility of a major chlor-alkali industry located in the area using expert elicitation and proven fuzzy logic. Sensitivity analysis has been done to evaluate the percentage contribution of each basic event that could lead to chlorine release. Two dimensional fuzzy fault tree analysis (TDFFTA) has been proposed for balancing the hesitation factor invo1ved in expert elicitation .

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In this thesis we have studied a few models involving self-generation of priorities. Priority queues have been extensively discussed in literature. However, these are situations involving priority assigned to (or possessed by) customers at the time of their arrival. Nevertheless, customers generating into priority is a common phenomena. Such situations especially arise at a physicians clinic, aircrafts hovering over airport running out of fuel but waiting for clearance to land and in several communication systems. Quantification of these are very little seen in literature except for those cited in some of the work indicated in the introduction. Our attempt is to quantify a few of such problems. In doing so, we have also generalized the classical priority queues by introducing priority generation ( going to higher priorities and during waiting). Systematically we have proceeded from single server queue to multi server queue. We also introduced customers with repeated attempts (retrial) generating priorities. All models that were analyzed in this thesis involve nonpreemptive service. Since the models are not analytically tractable, a large number of numerical illustrations were produced in each chapter to get a feel about the working of the systems.

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Parkinson's disease is a chronic progressive neurodegenerative movement disorder characterized by a profound and selective loss of nigrostriatal dopaminergic neurons. Our findings demonstrated that glutamatergic system is impaired during PD. The evaluations of these damages have important implications in understanding the molecular mechanism underlying motor, cognitive and memory deficits in PD. Our results showed a significant increase of glutamate content in the brain regions of 6- OHDA infused rat compared to control. This increased glutamate content caused an increase in glutamatergic and NMDA receptors function. Glutamate receptor subtypes- NMDAR1, NMDA2B and mGluR5 have differential regulatory role in different brain regions during PD. The second messenger studies confirmed that the changes in the receptor levels alter the IP3, cAMP and cGMP content. The alteration in the second messengers level increased the expression of pro-apoptotic factors - Bax and TNF-α, intercellular protein - α-synuclein and reduced the expression of transcription factor - CREB. These neurofunctional variations are the key contributors to motor and cognitive abnormalities associated with PD. Nestin and GFAP expression study confirmed that 5-HT and GABA induced the differentiation and proliferation of the BMC to neurons and glial cells in the SNpc of rats. We also observed that activated astrocytes are playing a crucial role in the proliferation of transplanted BMC which makes them significant for stem cell-based therapy. Our molecular and behavioural results showed that 5-HT and GABA along with BMC potentiates a restorative effect by reversing the alterations in glutamate receptor binding, gene expression and behaviour abnormality that occur during PD. The therapeutic significance in Parkinson’s disease is of prominence.

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The study was planned to investigate the bioactive compounds in Njavara compared to staple varieties and their bioactivity to substantiate the medicinal properties. Results of the study on chemical indices, antioxidant activity and antiinflammatory activity (in vivo) of Njavara black rice bran and rice in comparison with non-medicinal varieties like Sujatha and Palakkadan Matta rice bran and rice are given. The phytochemical investigation and quantification of Njavara extracts in comparison with staple varieties are detailed in this study. The last chapter is divided in three sections (A, B and C). Section A comprises the antioxidant activity by in vitro assays like DPPH, superoxide anion radical and hydrogen peroxide scavenging activity of the compounds. Also, theoretical studies using DFT were carried out based on DPPH radical scavenging activity for understanding the radical stability and mechanism of antioxidant activity. Section B comprises the anti-inflammatory activity of the identified compounds namely tricin and two flavonolignans in both in vivo and in vitro models. Section C describes the cytotoxicity of the rare flavonolignans, tricin 4’-O-(erythro-β-guaiacylglyceryl) ether and tricin 4’-O-(threo-β-guaiacylglyceryl) ether towards multiple cancer cells belonging to colon, ovarian and breast tumours.

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This thesis entitled Fish habitats and species assemblage in the selected rivers of kerala and investigation on life history traits of puntius carnaticus (JERDON,1849). Ecology is a new and exceedingly complex field of study, even though its concept was recognized by the Apostles in their use of the phrase ‘all flesh is grass.central role to play both in order to understand better the biodiversity phenomenon and to be able to draw up clear guidelines for careful resource management. In a review by WWF, IUCN and UNEP on the ways of conserving genetic diversity of freshwater fish it was recommended that the best way to conserve species diversity is to conserve habitat.The habitat studies in freshwater ecosystems are very essential for the proper understanding and management of human impact on fish diversity, to study the relationship between habitat variables and fish species assemblage structure, quantification of ecosystem degradation, habitat quality and biotic integrity of the ecosystems, development of habitat suitability index (I-ISI) models and classification of river reaches based on their physico-chemical properties. Therefore in the present study an attempt was made to assess the biodiversity potential and the relationship between habitat variables and fish species assemblage structure in six major river systems of Kerala which would be very useful in impressing upon the seriousness of habitat degradIn the present study, in Kabbini river system 15 locations encompassing between 721 946m above MSL were surveyed.ation and biotic devastation undergone in the major river systems of Kerala.During the present study the Habitat Quality Score (HQ) developed by the Ohio EPA was applied for the first time in India.The result of the present study revealed that, among various variables analysed, altitude has a very significant influence in deciding the fish diversity in six major river systems of Kerala. The fish diversity studied on the basis of Shanon-Weiner and Simpson diversity indices revealed that even though some minor variations occur with the suitability and complexity of habitats, the altitude showed inverse relationship with fish diversity.The present study revealed that the National Policy on the interlinking of rivers would permanently alter the HSI indices of the above mentioned fish species, which are now solely protected by the individuality of the rivers where their limited occurrence was notice.

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Ozone present in the atmosphere not only absorbs the biologically harmful ultraviolet radiation but also is an important ingredient of the climate system. The radiative absorption properties of ozone make it a determining factor in the structure of the atmosphere. Ozone in the troposphere has many negative impacts on humans and other living beings. Another significant aspect is the absorption of outgoing infrared radiation by ozone thus acting as a greenhouse gas. The variability of ozone in the atmosphere involves many interconnections with the incoming and outgoing radiation, temperature circulation etc. Hence ozone forms an important part of chemistry-climate as well as radiative transfer models. This aspect also makes the quantification of ozone more important. The discovery of Antarctic ozone hole and the role of anthropogenic activities in causing it made it possible to plan and implement necessary preventive measures. Continuous monitoring of ozone is also necessary to identify the effect of these preventive steps. The reactions involving the formation and destruction of ozone are influenced significantly by the temperature fluctuations of the atmosphere. On the other hand the variations in ozone can change the temperature structure of the atmosphere. Indian subcontinent is a region having large weather and climate variability which is evident from the large interannual variability of monsoon system over the region. Nearly half of Indian region comprises the tropical region. Most of ozone is formed in the tropical region and transported to higher latitudes. The formation and transport of ozone can be influenced by changes in solar radiation and various atmospheric circulation features. Besides industrial activities and vehicular traffic is more due to its large population. This may give rise to an increase in the production of tropospheric ozone which is greenhouse gas. Hence it becomes necessary to monitor the atmospheric ozone over this region. This study probes into the spatial distribution and temporal evolution of ozone over Indian subcontinent and discusses the contributing atmospheric parameters.

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At this era of energy crisis and resource depletion, availability of conventional materials throughout the year in quantity and quality, pose a hectic problem for the builders. Adding fuel to the fire, the demand of these materials increases day by day, since the housing and habitat requirements exponentially increase time to time. There is an international concern over this crisis and researchers are reorienting themselves, so as to evolve appropriate masonry units, using locally available cheap materials and technology. The concept of green material and construction has been well conceived in the research so that marginal materials and unskilled labour can be employed for the mass production of building blocks. In this context, considering earth as a sustainable material, there is a growing interest in the use of it, as a modern construction material. Solid waste management is one of the current major environmental concerns in our country. Our country is left with millions of cubic metre of waste plastics. One of the methods to satisfactorily address this solid waste management and the environmental issues is to suitably accommodate the waste in some form (as fibres). Their employability in block making in the form of fibres (plastic fibre- mud blocks) can be investigated through a fundamental research. Also, the review of the existing literature shows that most studies on natural fibres are focussed on cellulose based/ vegetable fibres obtained from renewable plant resources except in very few cases, where animal fibre, plastic fibre and polystyrene fabric were used. At this context, for the plastic fibre-mud blocks to be more widely applicable, a systematic quantification of the relevant physical and mechanical properties of the fibre masonry units is crucial, to enable an objective evaluation of the composite material’s response to actual field condition. This research highlights the salient observations from the detailed investigation of a systematic study on the effect of embedded fibres, made of plastic wastes on the performance of stabilised mud blocks.

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Roughness and defects induced on few-layer graphene (FLG) irradiated by Ar+ ions at different energies were investigated using X-ray photoemission spectroscopy (XPS) and atomic force microscopy techniques. The results provide direct experimental evidence of ripple formation, sp2 to sp3 hybridized carbon transformation, electronic damage, Ar+ implantation, unusual defects and edge reconstructions in FLG, which depend on the irradiation energy. In addition, shadowing effects similar to those found in oblique-angle growth of thin films were seen. Reliable quantification of the transition from the sp2-bonding to sp3-hybridized state as a result of Ar+ ion irradiation is achieved from the deconvolution of the XPS C (1s) peak. Although the ion irradiation effect is demonstrated through the shape of the derivative of the Auger transition C KVV spectra, we show that the D parameter values obtained from these spectra which are normally used in the literature fail to account for the sp2 to sp3 hybridization transition. In contrast to what is known, it is revealed that using ion irradiation at large FLG sample tilt angles can lead to edge reconstructions. Furthermore, FLG irradiation by low energy of 0.25 keV can be a plausible way of peeling graphene layers without the need of Joule heating reported previously