Search results for: polymerase chain reaction

Authors: Sasangka Prasetyawan, Anna Roosdiana, Diah Mardiana, Suratmo

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Pectinase are enzymes that hydrolyze pectin compounds. The use of this enzyme is primarily to reduce the viscosity of the beverage thus simplifying the purification process. Pectinase activity influenced by microbial sources . Exploration of two types of microbes that Aspergillus spp. and Bacillus spp. pectinase give different performance, but the use of local strain is still not widely studied. The aim of this research is exploration of pectinase from A. niger and B. firmus include production conditions and characterization. Bacillus firmus incubated and shaken at a speed of 200 rpm at pH variation (5, 6, 7, 8, 9, 10), temperature (30, 35, 40, 45, 50) °C and incubation time (6, 12, 18, 24, 30, 36 ) hours. Media was centrifuged at 3000 rpm, pectinase enzyme activity determined. Enzyme production by A. niger determined to variations in temperature and pH were similar to B. firmus, but the variation of the incubation time was 24, 48, 72, 96, 120 hours. Pectinase crude extract was further purified by precipitation using ammonium sulfate saturation in fraction 0-20 %, 20-40 %, 40-60 %, 60-80 %, then dialyzed. Determination of optimum conditions pectinase activity performed by measuring the variation of enzyme activity on pH (4, 6, 7, 8, 10), temperature (30, 35, 40, 45, 50) °C, and the incubation time (10, 20, 30, 40, 50) minutes . Determination of kinetic parameters of pectinase enzyme reaction carried out by measuring the rate of enzyme reactions at the optimum conditions, but the variation of the concentration of substrate (pectin 0.1 % , 0.2 % , 0.3 % , 0.4 % , 0.5 % ). The results showed that the optimum conditions of production of pectinase from B. firmus achieved at pH 7-8.0, 40-50 ⁰C temperature and fermentation time 18 hours. Purification of pectinase showed the highest purity in the 40-80 % ammonium sulfate fraction. Character pectinase obtained : the optimum working conditions of A. niger pectinase at pH 5 , while pectinase from B. firmus at pH 7, temperature and optimum incubation time showed the same value, namely the temperature of 50 ⁰C and incubation time of 30 minutes. The presence of metal ions can affect the activity of pectinase , the concentration of Zn 2 + , Pb 2 + , Ca 2 + and K + and 2 mM Mg 2 + above 6 mM inhibit the activity of pectinase .

Keywords: pectinase, Bacillus firmus, Aspergillus niger, green chemistry

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Authors: Latha Krishnan, Andrew Cobley

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Epoxy resin is most widely used as matrices for composites of aerospace, automotive and electronic applications due to its outstanding mechanical properties. These properties are chiefly predetermined by the chemical structure of the prepolymer and type of hardener but can also be varied by the processing conditions such as prepolymer and hardener mixing, degassing and curing conditions. In this research, the effect of degassing on the curing behaviour and the void occurrence is experimentally evaluated for epoxy /anhydride resin system. The epoxy prepolymer was mixed with an anhydride hardener and accelerator in an appropriate quantity. In order to investigate the effect of degassing on the curing behaviour and void content of the resin, the uncured resin samples were prepared using three different methods: 1) no degassing 2) degassing on prepolymer and 3) degassing on mixed solution of prepolymer and hardener with an accelerator. The uncured resins were tested in differential scanning calorimeter (DSC) to observe the changes in curing behaviour of the above three resin samples by analysing factors such as gel temperature, peak cure temperature and heat of reaction/heat flow in curing. Additionally, the completely cured samples were tested in DSC to identify the changes in the glass transition temperature (Tg) between the three samples. In order to evaluate the effect of degassing on the void content and morphology changes in the cured epoxy resin, the fractured surfaces of cured epoxy resin were examined under the scanning electron microscope (SEM). Also, the changes in the mechanical properties of the cured resin were studied by three-point bending test. It was found that degassing at different stages of resin mixing had significant effects on properties such as glass transition temperature, the void content and void size of the epoxy/anhydride resin system. For example, degassing (vacuum applied on the mixed resin) has shown higher glass transition temperature (Tg) with lower void content.

Keywords: anhydride epoxy, curing behaviour, degassing, void occurrence

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Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov

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The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.

Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction

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Authors: Maurizio E. Picarella, Ludovica Fumelli, Francesca Siligato, Andrea Mazzucato

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Studies on reproductive dynamics in crops subjected to heat stress are crucial to breed more tolerant cultivars. In tomato, cultivars, breeding lines, and wild species have been thoroughly evaluated for the response to heat stress in several studies. Here, we address the reaction to temperature stress in a panel of selected landraces representing genotypes cultivated before the advent of professional varieties that usually show high adaptation to local environments. We adopted an experimental design with two open field trials, where transplanting was spaced by one month. In the second field, plants were thus subjected to mild stress with natural temperature fluctuations. The genotypes showed wide variation for both vegetative (plant height) and reproductive (stigma exsertion, pollen viability, number of flowers per inflorescence, and fruit set) traits. On average, all traits were affected by heat conditions; except for the number of flowers per inflorescence, the “G*E” interaction was always significant. In agreement with studies based on different materials, estimated broad sense heritability was high for plant height, stigma exsertion, and pollen viability and low for the number of flowers per inflorescence and fruit set. Despite the interaction, traits recorded in control and in heat conditions were positively correlated. The first two principal components estimated by multivariate analysis explained more than 50% of the total variability. The study indicated that landraces present a wide variability for the response of reproductive traits to temperature stress and that such variability could be very informative to dissect the traits with higher heritability and identify new QTL useful for breeding more resilient varieties.

Keywords: fruit set, heat stress, solanum lycopersicum L., style exsertion, tomato

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Authors: Dhara Shah, Chandrakant Shah

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Construction industry utilizes plenty of water in the name of curing. Looking at the present scenario, the days are not so far when all construction industries will have to switch over to an alternative-self curing system, not only to save water for sustainable development of the environment but also to promote indoor and outdoor construction activities even in water scarce areas. At the same time, curing is essential for the development of proper strength and durability. IS 456-2000 recommends a curing period of 7 days for ordinary Portland cement concrete, and 10 to 14 days for concrete prepared using mineral admixtures or blended cements. But, being the last act in the concreting operations, it is often neglected or not fully done. Consequently, the quality of hardened concrete suffers, more so, if the freshly laid concrete gets exposed to the environmental conditions of low humidity, high wind velocity and high ambient temperature. To avoid the adverse effects of neglected or insufficient curing, which is considered a universal phenomenon, concrete technologist and research scientists have come up with curing compounds. Concrete is said to be self-cured, if it is able to retain its water content to perform chemical reaction for the development of its strength. Curing compounds are liquids which are either incorporated in concrete or sprayed directly onto concrete surfaces and which then dry to form a relatively impermeable membrane that retards the loss of moisture from the concrete. They are an efficient and cost-effective means of curing concrete and may be applied to freshly placed concrete or that which has been partially cured by some other means. However, they may affect the bond between concrete and subsequent surface treatments. Special care in the choice of a suitable compound needs to be exercised in such circumstances. Curing compounds are generally formulated from wax emulsions, chlorinated rubbers, synthetic and natural resins, and from PVA emulsions. Their effectiveness varies quite widely, depending on the material and strength of the emulsion.

Keywords: curing methods, self-curing compound, compressive strength, modulus of elasticity, durability

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Authors: N. V. Kazmiruk, Y. R. Nartsissov

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Phosphate activated glutaminase (GA, E.C. 3.5.1.2) plays a key role in glutamine/glutamate homeostasis in mammalian brain, catalyzing the hydrolytic deamidation of glutamine to glutamate and ammonium ions. GA is mainly localized in mitochondria, where it has the catalytically active form on the inner mitochondrial membrane (IMM) and the other soluble form, which is supposed to be dormant. At present time, the exact localization of the membrane glutaminase active site remains a controversial and an unresolved issue. The first hypothesis called c-side localization suggests that the catalytic site of GA faces the inter-membrane space and products of the deamidation reaction have immediate access to cytosolic metabolism. According to the alternative m-side localization hypothesis, GA orients to the matrix, making glutamate and ammonium available for the tricarboxylic acid cycle metabolism in mitochondria directly. In our study, we used a multi-compartment kinetic approach to simulate metabolism of glutamate and glutamine in the astrocytic cytosol and mitochondria. We used physiologically important ratio between the concentrations of glutamine inside the matrix of mitochondria [Glnₘᵢₜ] and glutamine in the cytosol [Glncyt] as a marker for precise functioning of the system. Since this ratio directly depends on the mitochondrial glutamine carrier (MGC) flow parameters, key observation was to investigate the dependence of the [Glnmit]/[Glncyt] ratio on the maximal velocity of MGC at different initial concentrations of mitochondrial glutamate. Another important task was to observe the similar dependence at different inhibition constants of the soluble GA. The simulation results confirmed the experimental c-side localization hypothesis, in which the glutaminase active site faces the outer surface of the IMM. Moreover, in the case of such localization of the enzyme, a 3-fold decrease in ammonium production was predicted.

Keywords: glutamate metabolism, glutaminase, kinetic approach, mitochondrial membrane, multi-compartment modeling

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Authors: Sunantha Teyarachakul

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We consider the worker-learning Newsvendor Model, under the case of lost-sales for unmet demand, with the research objective of proposing the cost-minimization order policy and lot size, scheduled to arrive at the beginning of the selling-period. In general, the New Vendor Model is used to find the optimal order quantity for the perishable items such as fashionable products or those with seasonal demand or short-life cycles. Technically, it is used when the product demand is stochastic and available for the single selling-season, and when there is only a one time opportunity for the vendor to purchase, with possibly of long ordering lead-times. Our work differs from the classical Newsvendor Model in that we incorporate the human factor (specifically worker learning) and its influence over the costs of processing units into the model. We describe this by using the well-known Wright’s Learning Curve. Most of the assumptions of the classical New Vendor Model are still maintained in our work, such as the constant per-unit cost of leftover and shortage, the zero initial inventory, as well as the continuous time. Our problem is challenging in the way that the best order quantity in the classical model, which is balancing the over-stocking and under-stocking costs, is no longer optimal. Specifically, when adding the cost-saving from worker learning to such expected total cost, the convexity of the cost function will likely not be maintained. This has called for a new way in determining the optimal order policy. In response to such challenges, we found a number of characteristics related to the expected cost function and its derivatives, which we then used in formulating the optimal ordering policy. Examples of such characteristics are; the optimal order quantity exists and is unique if the demand follows a Uniform Distribution; if the demand follows the Beta Distribution with some specific properties of its parameters, the second derivative of the expected cost function has at most two roots; and there exists the specific level of lot size that satisfies the first order condition. Our research results could be helpful for analysis of supply chain coordination and of the periodic review system for similar problems.

Keywords: inventory management, Newsvendor model, order policy, worker learning

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Authors: Hajar Zarei, AliAkbar Zarenejadatashgah, Vuk Uskoković, Hiroshi Watabe

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The reactive oxygen species (ROS) generated by radiation in nuclear diagnostic imaging and radiotherapy could damage the structure of the proteins in noncancerous cells surrounding the tumor. The critical factor in many age-related diseases, such as Alzheimer, Parkinson, or Huntington diseases, is the oxidation of proteins by the ROS as molecular triggers of the given pathologies. Our studies by spectroscopic experiments showed doses close to therapeutic ones (1 to 5 Gy) could lead to changes of secondary and tertiary structures in BSA protein macromolecule as a protein model as well as the aggregation of polypeptide chain but without the fragmentation. For this reason, we investigated the radioprotective effects of natural (vitamin C and E) and synthetic materials (CNPs and FIOMPs) on the structural changes in BSA protein induced by gamma irradiation at a therapeutic dose (3Gy). In the presence of both vitamins and synthetic materials, the spectroscopic studies revealed that irradiated BSA was protected from the structural changes caused by ROS, according to in vitro research. The radioprotective property of CNPs and FIOMPs arises from enzyme mimetic activities (catalase, superoxide dismutase, and peroxidase) and their antioxidant capability against hydroxyl radicals. In the case of FIOMPs, a porous structure also leads to increased ROS recombination with each other in the same radiolytic track and subsequently decreased encounters with BSA. The hydrophilicity of vitamin C resulted in the major scavenging of ROS in the solvent, whereas hydrophobic vitamin E localized on the nonpolar patches of the BSA surface, where it did not only neutralize them thanks to the moderate BSA binding constant but also formed a barrier for diffusing ROS. To the best of our knowledge, there has been a persistent lack of studies investigating the radioactive effect of mentioned materials on proteins. Therefore, the results of our studies can open a new widow for application of these common dietary ingredients and new synthetic NPs in improving the safety of radiotherapy.

Keywords: reactive oxygen species, spectroscopy, bovine serum albumin, gamma radiation, radioprotection

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Authors: R. M. S. Sachini Amararathne

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This research is striving to develop a thermo degradable starch plastic compound/ masterbatch for industrial packaging applications. A native corn starch-modified with an esterification reaction of lauric acid is melt blent with an unsaturated elastomer (styrene-butadiene-rubber/styrene-butadiene-styrene). A trace amount of metal salt is added into the internal mixer to study the effect of pro-oxidants in a thermo oxidative environment. Then the granulated polymer composite which is consisted with 80-86% of polyolefin (LLDP/LDPE/PP) as the pivotal agent; is extruded with processing aids, antioxidants and some other additives in a co-rotating twin-screw extruder. The pelletized composite is subjected to compression molding/ Injection molding or blown film extrusion processes to acquire the samples/specimen for tests. The degradation process is explicated by analyzing the results of fourier transform infrared spectroscopy (FTIR) measurements, thermo oxidative aging studies (placing the dumb-bell specimen in an air oven at 70 °C for four weeks of exposure.) governed by tensile and impact strength test reports. Furthermore, the samples were elicited into manifold outdoors to inspect the degradation process. This industrial process is implemented to reduce the volume of fossil-based garbage by achieving the biodegradability and compostability in the natural cycle. Hence the research leads to manufacturing a degradable plastic packaging compound which is now available in the Sri Lankan market.

Keywords: blown film extrusion, compression moulding, polyolefin, pro-oxidant, styrene-butadine-rubber, styrene-butadiene-styrene, thermo oxidative aging, unsaturated elastomer

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Authors: Milad Kurdi, Hadi Farhadian, Teymour Eslamkish

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This study examined the hydro-geochemistry of Qare-Sou catchment and Gorgan Gulf in order to determine the spatial distribution of major ions. In this regard, six hydrometer stations in the catchment and four stations in Gorgan Gulf were chosen and the samples were collected. Results of spatial and temporal distribution of major ions have shown similar variation trends for calcium, magnesium, and bicarbonate ions. Also, the spatial trend of chloride, sulfate, sodium and potassium ions were same as Electrical Conductivity (EC) and Total Dissolved Solid (TDS). In Nahar Khoran station, the concentrations of ions were more than other stations which may be related to human activities and the role of geology. The Siah Ab station’s ions showed high concentration which is may be related to the station’s close proximity to Gorgan Gulf and the return of water to Qare-Sou River. In order to determine the interaction of water and rock, the Gibbs diagram was used and the results showed that water of the river falls in the rock range and it is affected more by weathering and reaction between water and stone and less by evaporation and crystallization. Assessment of the quality of river water by using graphic methods indicated that the type of water in this area is Ca-HCO₃-Mg. Major ions concentration in Qare-Sou in the universal average was more than but not more than the allowed limit by the World Health Organization and China Standard Organization. A comparison of ions concentration in Gorgan Gulf, seas and oceans showed that the pH in Gorgan Gulf was more than the other seas but in Gorgan Gulf the concentration of anion and cation was less than other seas.

Keywords: hydro-geochemistry, Qare-Sou river, Gorgan gulf, major ions, Gibbs diagram, water quality, graphical methods

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Authors: Charlotte Williams

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Yale University Professor Hiram Bingham’s discovery of Machu Picchu in 1911 spurred an international interest in the Inca Empire, and with it, a dispute with the Peruvian government over who had rightful jurisdiction and curatorship over Inca history. By 2011, the Peruvian government initiated a legal battle for the return of artifacts that Bingham had removed from Machu Picchu, successfully returning them not to the site of Machu Picchu, but to Cusco, employing the rationale that the ancient Inca capital housed descendants of the Inca empire. This conflation of the past and present can be traced to a largely unanalyzed study that accompanied Bingham’s expedition: an ethnographic analysis of Inca descendants, which at the time portrayed indigenous Peruvian Andean peoples as remnants of a lost civilization, using Cusco as an assumed repository for people with 'Inca' characteristics. This study draws from the original Yale Peruvian Expedition archives, the Cusco Library archives, and in-depth interviews with curators of the Inca Museum and Machu Picchu Museum to analyze both the political conflict that emerged as a reaction to the ethnographic study, and how the study articulated with an inflating tourism market attempting to define what it meant to be Inca to an international public. The construction of the modern Inca as both directors of tourism management and purveyors of their archaeological material culture points to a unique case in which modern Peruvian citizens could claim heritage to an Inca past despite a lack of recognition as a legally defined group. The result has far-reaching implications, since Bingham’s artifacts returned not necessarily to a traditional nation-state, but to an imagined one, broadening the conditions under which informal repatriations can occur.

Keywords: archaeology of memory, imagined communities, Incanismo, repatriation

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Authors: Zahra F. Al-Khateeb, Shenel Shekerzade, Hasna Boumenar, Siân M. Henson, Jordi L. Tremoleda, A. T. Michael-Titus

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Traumatic brain injury (TBI) is the leading cause of disability and death in young adults and also increases the risk ofneurodegeneration. The mechanisms linking moderate to severe TBI to neurodegeneration are not known. It has been proposed that cellular senescence inductionpost-injury could amplify neuroinflammation and induce long-term changes. The impact of these processes after injury to an immature brain has not been characterised yet. We carried out a controlled cortical impact injury (CCI) in juvenile 1 month-old male CD1 mice. Animals were anesthetised and received a unilateral CCI injury. The sham group received anaesthesia and had a craniotomy. A naïve group had no intervention. The brain tissue was analysed at 5 days and 35 days post-injury using immunohistochemistry and markers for microglia, astrocytes, and senescence. Compared tonaïve animals, injured mice showed an increased microglial and astrocytic reaction early post-injury, as reflected in Iba1 and GFAP markers, respectively; the GFAP increase persisted in the later phase. The senescence analysis showed a significant increase inγH2AX-53BP1 nuclear foci, 8-oxoguanine, p19ARF, p16INK4a, and p53 expression in naïve vs. sham groups and naïve vs. CCI groups, at 5 dpi. At 35 days, the difference was no longer statistically significant in all markers. The injury induced a decrease p21 expression vs. the naïve group, at 35 dpi. These results indicate the induction of a complex senescence response after immature brain injury. Some changes occur early and may reflect the activation/proliferation of non-neuronal cells post-injury that had been hindered, whereas changes such as p21 downregulation may reflect a delayed response and pro-repair processes.

Keywords: cellular senescence, traumatic brain injury, brain injury, controlled cortical impact

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Authors: Tuba Zahra, Aakash Parekh

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Hyperspectral remote sensing refers to imaging of objects or materials in narrow conspicuous spectral bands. Hyperspectral images (HSI) enable the extraction of spectral signatures for objects or materials observed. These images contain information about the reflectance of each pixel across the electromagnetic spectrum. It enables the acquisition of data simultaneously in hundreds of spectral bands with narrow bandwidths and can provide detailed contiguous spectral curves that traditional multispectral sensors cannot offer. The contiguous, narrow bandwidth of hyperspectral data facilitates the detailed surveying of Earth's surface features. This would otherwise not be possible with the relatively coarse bandwidths acquired by other types of imaging sensors. Hyperspectral imaging provides significantly higher spectral and spatial resolution. There are several use cases that represent the commercial applications of hyperspectral remote sensing. Each use case represents just one of the ways that hyperspectral satellite imagery can support operational efficiency in the respective vertical. There are some use cases that are specific to VNIR bands, while others are specific to SWIR bands. This paper discusses the different commercially viable use cases that are significant for HSI application areas, such as agriculture, mining, oil and gas, defense, environment, and climate, to name a few. Theoretically, there is n number of use cases for each of the application areas, but an attempt has been made to streamline the use cases depending upon economic feasibility and commercial viability and present a review of literature from this perspective. Some of the specific use cases with respect to agriculture are crop species (sub variety) detection, soil health mapping, pre-symptomatic crop disease detection, invasive species detection, crop condition optimization, yield estimation, and supply chain monitoring at scale. Similarly, each of the industry verticals has a specific commercially viable use case that is discussed in the paper in detail.

Keywords: agriculture, mining, oil and gas, defense, environment and climate, hyperspectral, VNIR, SWIR

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Authors: Belahmadi M. S. O., Abdessemed A., Benchiheub M., Doukali H., Kaid Kasbah K. M.

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The aim of this study is to evaluate the quality of leachate generated by the Batna landfill site, and to verify the performance of various advanced oxidation processes, in particular the Fenton and Photo-Fenton systems combined with biological treatment to eliminate the recalcitrant organic matter contained in this effluent, and to preserve reverse osmosis membranes used for leachate treatment. The average values obtained are compared with national and international discharge standards. The results of physico-chemical analyses show that the leachate has an alkaline pH =8.26 and a high organic load with a low oxygen content. Mineral pollution is represented by high conductivity (38.3 mS/cm), high Kjeldahl nitrogen content (1266.504 mg/L) and ammoniacal nitrogen (1098.384 mg/L). The average pollution indicator parameters measured were: BOD5 = 1483.333 mg O2 /L, COD = 99790.244 mg O 2/L, TOC = 22400 mg C/L. These parameters exceed Algerian standards. Hence, there is a necessity to treat this effluent before discharging it into the environment. A comparative study was carried out to estimate the efficiency of two oxidation processes. Under optimum reaction conditions, TOC removal efficiencies of 63.43% and 73.4% were achieved for the Fenton and Photo-Fenton processes, respectively. COD removal rates estimated at 88% and 99.5% for the Fenton and Photo- Fenton processes, respectively. In addition, the Photo-Fenton + bacteria + micro- algae hybrid treatment gave removal efficiencies of around 92.24% for TOC and 99.9% for COD; -0.5 for AOS and 0.01 for CN. The results obtained during this study showed that a hybrid approach combining the PhotoFenton process and biological treatment appears to be a highly effective alternative for achieving satisfactory treatment, which aimed at exploiting the advantages of this method in terms of organic pollutant removal.

Keywords: leachate, landfill, advanced oxidation processes, Fenton and Photo-Fenton systems, biological treatment, organic pollutants

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Authors: Bianca W. O. Silva, Adriana C. M. Andrade, Gustavo C. M. Lucena, Virna M. S. Lima

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Introduction: Basic life support (BLS) involves the immediate recognition of cardiopulmonary arrest. Each year, 359.400 and 275.000 individuals with cardiac arrest are attended in emergency departments in USA and Europe. Brazilian data shows that 200.000 cardiac arrests occur every year, and half of them out of the hospital. Medical schools around the world teach BLS in the first years of the course, but studies show that there is a decline of the knowledge as the years go by, affecting the chain of survival. The objective was to analyze the knowledge of medical students about BLS and the retention of this learning throughout the course. Methods: This study included 150 students who were at the second and third year of a medical school in Salvador, Bahia, Brazil. The instrument of data collection was a structured questionnaire composed of 20 questions based on the 2015 American Heart Association guideline. The Pearson Chi-square test was used in order to study the association between previous training, sex and semester with the degree of knowledge of the students. The Kruskal-Wallis test was used to evaluate the different yields obtained between the various semesters. The number of correct answers was described by average and quartiles. Results: Regarding the degree of knowledge, 19.6% of the female students reached the optimal classification, a better outcome than the achieved by the male participants. Of those with previous training, 33.33% were classified as good and optimal, none of the students reached the optimal classification and only 2.2% of them were classified as bad (those who did not have 52.6% of correct answers). The analysis of the degree of knowledge related to each semester revealed that the 5th semester had the highest outcome: 30.5%. However, the acquaintance presented by the semesters was generally unsatisfactory, since 50% of the students, or more, demonstrated knowledge levels classified as bad or regular. When confronting the different semesters and the achieved scores, the value of p was 0.831. Conclusion: It is important to focus on the training of medical professionals that are capable of facing emergency situations, improving the systematization of care, and thereby increasing the victims' possibility of survival.

Keywords: basic life support, cardiopulmonary ressucitacion, education, medical students

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Authors: Narasamma Nippatlapallia

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Communities across the world are desperate to get their environment free of toxic per-poly fluoroalkyl substances (PFAS) especially when these chemicals are in aqueous media. In the present study, two different chain length PFAS (PFHxA (C6), PFDA (C10)) are selected for degradation using a modified continuous flow nonthermal plasma. The results showed 82.3% PFHxA and 94.1 PFDA degradation efficiencies, respectively. The defluorination efficiency is also evaluated which is 28% and 34% for PFHxA and PFDA, respectively. The results clearly indicates that the structure of PFAS has a great impact on degradation efficiency. The effect of flow rate is studied. increase in flow rate beyond 2 mL/min, decrease in degradation efficiency of the targeted PFAS was noticed. PFDA degradation was decreased from 85% to 42%, and PFHxA was decreased to 32% from 64% with increase in flow rate from 2 to 5 mL/min. Similarly, with increase in flow rate the percentage defluorination was decreased for both C10, and C6 compounds. This observation can be attributed to mainly because of change in residence time (contact time). Real water/wastewater is a composition of various organic, and inorganic ions that may affect the activity of oxidative species such as 𝑂𝐻. radicals on the target pollutants. Therefore, it is important to consider radicals quenching chemicals to understand the efficiency of the reactor. In gas-liquid NTP discharge reactors 𝑂𝐻. , 𝑒𝑎𝑞 − , 𝑂 . , 𝑂3, 𝐻2𝑂2, 𝐻. are often considered as reactive species for oxidation and reduction of pollutants. In this work, the role played by two distinct 𝑂 .𝐻 Scavengers, ethanol and glycerol, on PFAS percentage degradation, and defluorination efficiency (i,e., fluorine removal) are measured was studied. The addition of scavenging agents to the PFAS solution diminished the PFAS degradation to different extents depending on the target compound molecular structure. In comparison with the degradation of only PFAS solution, the addition of 1.25 M ethanol inhibited C10, and C6 degradation by 8%, and 12%, respectively. This research was supported with energy efficiency, production rate, and specific yield, fluoride, and PFAS concentration analysis with respect to optimum hydraulic retention time (HRT) of the continuous flow reactor.

Keywords: wastewater, PFAS, nonthermal plasma, mineralization, defluorination

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Authors: Maciej Jedrzejczyk, Karolina Marzantowicz

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Traditional utility value chain is being transformed during last few years into unbundled markets. Increased distributed generation of energy is one of considerable challenges faced by Smart Grid networks. New sources of energy introduce volatile demand response which has a considerable impact on traditional middlemen in E&U market. The purpose of this research is to search for ways to allow near-real-time electricity markets to transact with surplus energy based on accurate time synchronous measurements. A proposed framework evaluates the use of secure peer-2-peer (P2P) communication and distributed transaction ledgers to provide flat hierarchy, and allow real-time insights into present and forecasted grid operations, as well as state and health of the network. An objective is to achieve dynamic grid operations with more efficient resource usage, higher security of supply and longer grid infrastructure life cycle. Methods used for this study are based on comparative analysis of different distributed ledger technologies in terms of scalability, transaction performance, pluggability with external data sources, data transparency, privacy, end-to-end security and adaptability to various market topologies. An intended output of this research is a design of a framework for safer, more efficient and scalable Smart Grid network which is bridging a gap between traditional components of the energy network and individual energy producers. Results of this study are ready for detailed measurement testing, a likely follow-up in separate studies. New platforms for Smart Grid achieving measurable efficiencies will allow for development of new types of Grid KPI, multi-smart grid branches, markets, and businesses.

Keywords: autonomous agents, Distributed computing, distributed ledger technologies, large scale systems, micro grids, peer-to-peer networks, Self-organization, self-stabilization, smart grids

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Authors: Vivek Thorat, Suhasini Desai

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The hybrid electric vehicle is widely accepted as a promising short to mid-term technical solution due to noticeably improved efficiency and low emissions at competitive costs. Retro fitment of hybrid components into a conventional vehicle for achieving better performance is the best solution so far. But retro fitment includes major modifications into a conventional vehicle with a high cost. This paper focuses on the development of a P3x hybrid prototype with rear wheel drive parallel hybrid electric Light Commercial Vehicle (LCV) with minimum and low-cost modifications. This diesel Hybrid LCV is different from another hybrid with regard to the powertrain. The additional powertrain consists of continuous contact helical gear pair followed by chain and sprocket as a coupler for traction motor. Vehicle powertrain which is designed for the intended high-speed application. This work focuses on targeting of design, development, and packaging of this unique parallel diesel-electric vehicle which is based on multimode hybrid advantages. To demonstrate the practical applicability of this transmission with P3x hybrid configuration, one concept prototype vehicle has been build integrating the transmission. The hybrid system makes it easy to retrofit existing vehicle because the changes required into the vehicle chassis are a minimum. The additional system is designed for mainly five modes of operations which are engine only mode, electric-only mode, hybrid power mode, engine charging battery mode and regenerative braking mode. Its driving performance, fuel economy and emissions are measured and results are analyzed over a given drive cycle. Finally, the output results which are achieved by the first vehicle prototype during experimental testing is carried out on a chassis dynamometer using MIDC driving cycle. The results showed that the prototype hybrid vehicle is about 27% faster than the equivalent conventional vehicle. The fuel economy is increased by 20-25% approximately compared to the conventional powertrain.

Keywords: P3x configuration, LCV, hybrid electric vehicle, ROMAX, transmission

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Authors: Sally A. El Awdan

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Objective and Design: The purpose of this study was to evaluate the anti inflammatory, anti-arthritic and analgesic effects of antidepressants. Methods: Carrageenan model was used to assess effect on acute inflammation. Paw volume were measured at 1, 2, 3 and 4th hour post challenge. Anti-nociceptive effect was evaluated by hot plate method. Chronic inflammation was developed using Complete Freund's Adjuvant (CFA). The animals were injected with Freund’s adjuvant in sub-plantar tissue of the right posterior paw. Paw volume, ankle flexion scores, adjuvant-induced hyperalgesia and serum cytokine levels were assessed. Results: Results obtained demonstrate that mirtazapine, venalfaxine and escitalopram significantly and dose-dependently inhibited carrageenan-induced rat paw oedema. Mirtazapine, venalfaxine and escitalopram increased the reaction time of rats in hot plate test. We observed an increase in paw volume, ankle flexion scores, thermal hyperalgesia, serum levels of interleukin-1β, PGE2 and TNF-α, induced by intraplantar CFA injection. Regular treatment up to 28 days of adjuvant-induced arthritic rats with mirtazapine, venalfaxine and escitalopram showed anti anti-inflammatory and analgesic activities by suppressing the paw volume, recovering the paw withdrawal latency, and by inhibiting the ankle flexion scores in CFA-induced rats. In addition significant reduction in serum levels of interleukin-1β, PGE2 and TNF-α level in arthritic rats was reduced by treatment with drugs. Conclusion: These results suggest that antidepressants have significant anti-inflammatory and anti-nociceptive effects in acute and chronic models in rats, which may be associated with the reduction of interleukin-1β, PGE2 and TNF-α levels.

Keywords: antidepressants, carrageenan, anti-nociceptive, Complete Freund's Adjuvant

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Authors: Unsain Dianne, Roussel Audrey, Goude Gwenaëlle, Magniez Pierre, Storå Jan

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This paper aims to investigate the changes in husbandry practices and meat consumption during the transition from the Medieval Climate Anomaly to the Little Ice Age in the South of France. More precisely, we will investigate breeding strategies, animal size and health status, carcass exploitation strategies, and the impact of socioeconomic status on human-environment interactions. For that purpose, we will analyze faunal remains from ten sites equally distributed between the two periods. Those include consumers from different socio-economic backgrounds (peasants, city dwellers, soldiers, lords, and the Popes). The research will employ different methods used in zooarchaeology: comparative anatomy, biometry, pathologies analyses, traceology, and utility indices, as well as experimental archaeology, to reconstruct and understand the changes in animal breeding and consumption practices. Their analysis will allow the determination of modifications in the animal production chain, with the composition of the flocks (species, size), their management (age, sex, health status), culinary practices (strategies for the exploitation of carcasses, cooking, tastes) or the importance of trade (butchers, sales of processed animal products). The focus will also be on the social extraction of consumers. The aim will be to determine whether climate change has had a greater impact on the most modest groups (such as peasants), whether the consequences have been global and have also affected the highest levels of society, or whether the social and economic factors have been sufficient to balance out the climatic hazards, leading to no significant changes. This study will contribute to our understanding of the impact of climate change on breeding and consumption strategies in medieval society from a historical and social point of view. It combines various research methods to provide a comprehensive analysis of the changes in human-animal interactions during different climatic periods.

Keywords: archaeology, animal economy, cooking, husbandry practices, climate change, France

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Authors: Dharmendra Pratap

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Large Cardamom (Amomum subulatum), family Zingiberaceae is an aromatic spice crop and has rich medicinal value. Large Cardamom is as synonymous to Sikkim as Tea is to Darjeeling. Since Sikkim alone contributes up to 88% of India's large cardamom production which is the world leader by producing over 50% of the global yield. However, the production of large cardamom has declined almost to half since last two decade. The economic losses have been attributed to two viral diseases namely, chirke and Foorkey. Chirke disease is characterized by light and dark green streaks on leaves. The affected leaves exhibit streak mosaic, which gradually coalesce, turn brown and eventually dry up. Excessive sprouting and formation of bushy dwarf clumps at the base of mother plants that gradually die characterize the foorkey disease. In our surveys in Sikkim–Darjeeling hill area during 2012-14, 40-45% of plants were found to be affected with foorkey disease and 10-15% with chirke. Mechanical and aphid transmission study showed banana as an alternate host for both the disease. For molecular identification, total genomic DNA and RNA was isolated from the infected leaf tissues and subjected to Rolling circle amplification (RCA) and RT-PCR respectively. The DNA concatamers produced in the RCA reaction were monomerized by different restriction enzymes and the bands corresponding to ~1 kb genomes were purified and cloned in the respective sites. The nucleotide sequencing results revealed the association of Nanovirus with the foorkey disease of large cardamom. DNA1 showed 74% identity with Replicase gene of FBNYV, DNA2 showed 77% identity with the NSP gene of BBTV and DNA3 showed 74% identity with CP gene of BBTV. The finding suggests the presence of a new species of nanovirus associated with foorkey disease of large cardamom in Sikkim and Darjeeling hills. The details of their epidemiology and other factors would be discussed.

Keywords: RCA, nanovirus, large cardamom, molecular virology and microbiology

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Authors: Alvaro Armenta-Ramade, Arturo Serrano-Santoyo, Veronica Rojas-Mendizabal, Roberto Conte-Galvan

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The availability and affordability of commercial off-the-shell products have brought a major impetus in the development of university projects related to the design, construction and launching of small satellites on a global scale. Universities in emerging economies as well as in least developed countries have been able to develop prototypes of small satellites (cubesats and cansats) with limited budgets. The experience gained in the development of small satellites gives rise to capacity building for designing more complex aerospace systems. This trend has significantly increased the pace and number of aerospace university projects around the world. In the case of Mexico, projects funded by different agencies have been very effective in accelerating the capacity building and technology transfer initiatives in the aerospace ecosystem. However, many of this initiatives have centered their efforts in technology development matters with minimum or no considerations of key regulatory issues related to frequency assignment, management and licensing, as well as launching requirements and measures of mitigation of space debris. These regulatory concerns are fundamental to accomplish successful missions that take into account the complete value chain of an aerospace project. The purpose of this paper is to develop a regulatory framework to support the efforts of educational institutions working on the development of small satellites in Mexico. We base our framework on recommendations from the International Telecommunications Union (ITU), the United Nations Office for Outer Space Affairs (UNOOSA) and other major actors of the Mexican regulatory ecosystem. In order to develop an integrated and cohesive framework, we draw on complexity science to identify the agents, their role and interactions. Our goal is to create a guiding instrument available both in print and online that can also be used in other regions of the world

Keywords: capacity building, complexity science, cubesats, space regulations, small satellites

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Authors: Federica Laface, Cristina Pedà, Francesco Longo, Francesca de Domenico, Riccardo Minichino, Pierpaolo Consoli, Pietro Battaglia, Silvestro Greco, Teresa Romeo

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Plastics pollution represents one of the most important threats to marine biodiversity. In the last decades, different species are investigated to evaluate the extent of the plastic ingestion phenomenon. Even if the cephalopods play an important role in the food chain, they are still poorly studied. The aim of this research was to investigate the plastic ingestion in two commercial cephalopod species from the southern Tyrrhenian Sea: the common octopus, Octopus vulgaris (n=6; mean mantle length ML 10.7 ± 1.8) and the common cuttlefish, Sepia officinalis (n=13; mean ML 13.2 ± 1.7). Plastics were extracted from the filters obtained by the chemical digestion of cephalopods gastrointestinal tracts (GITs), using 10% potassium hydroxide (KOH) solution in a 1:5 (w/v) ratio. Once isolated, particles were photographed, measured, and their size class, shape and color were recorded. A total of 81 items was isolated from 16 of the 19 examined GITs, representing a total occurrence (%O) of 84.2% with a mean value of 4.3 ± 8.6 particles per individual. In particular, 62 plastics were found in 6 specimens of O. vulgaris (%O=100) and 19 particles in 10 S. officinalis (%O=94.7). In both species, the microplastics size class was the most abundant (93.8%). Plastic items found in O. vulgaris were mainly fibers (61%) while fragments were the most frequent in S. officinalis (53%). Transparent was the most common color in both species. The analysis will be completed by Fourier transform infrared (FT-IR) spectroscopy technique in order to identify polymers nature. This study reports preliminary data on plastic ingestion events in two cephalopods species and represents the first record of plastic ingestion by the common octopus. Microplastic items detected in both common octopus and common cuttlefish could derive from secondary and/or accidental ingestion events, probably due to their behavior, feeding habits and anatomical features. Further studies will be required to assess the effect of marine litter pollution in these ecologically and commercially important species.

Keywords: cephalopods, GIT analysis, marine pollution, Mediterranean sea, microplastics

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Authors: P. Saha, R. Ganguly, N. K .Singha, A. Pich

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Microgel, a smart class of material, has drawn attention in the past few years due to its response to external stimuli like temperature, pH, and ionic strength of the solution. Among them, one type of polymer becomes soluble, and the other becomes insoluble in water upon heating displaying upper critical solution temperature (UCST) (e.g., polysulfobetaine, PSB) and lower critical solution temperature (LCST) (e.g., poly(N-vinylcaprolactam, PVCL)) respectively. Polyzwitterions, electrically neutral polymers are biocompatible, biodegradable, and non-cytotoxic in nature, and presence of zwitterionic pendant group in the main backbone makes them stable against temperature and pH variations and strong hydration capability in salt solution promotes them to be used as interfacial bio-adhesion resistance material. Majority of zwitterionic microgels have been synthesized in mini- emulsion technique using free radical polymerization approach. Here, a new route to synthesize dual thermo-responsive PVCL microgels decorated with appreciable amount of zwitterionic PSB chains was developed by a purely water-based surfactant-free reversible addition–fragmentation chain transfer (RAFT) precipitation polymerization. PSB macro-RAFTs having different molecular weights were synthesized and utilized for surface-grafting with PVCL microgels varying the macro-RAFT concentration using N,N′-methylenebis(acrylamide) (BIS) as cross-linker. Increasing the PSB concentration in the PVCL microgels resulted in a linear increase in UCST but decrease in hydrodynamic radius due to strong intrachain coulombic attraction forces acting between the opposite charges present in the zwitterionic groups. Anti- fouling effect was observed on addition of BSA protein solution on the microgel-coated membrane surfaces as studied by fluorescence spectrophotoscopy.

Keywords: microgels, polyzwitterions, upper critical solution temperature-lower critical solution temperature, UCST-LCST, ionic crosslinking

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Authors: Nathenal Thomas

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In the food chain and in many other agricultural products, corn, also known as maize, which goes by the scientific name Zea mays subsp, is a widely produced agricultural product. Corn has the highest adaptability. It comes in many different types, is employed in many different industrial processes, and is more adaptable to different agro-climatic situations. In Ethiopia, maize is among the most widely grown crop. Small-scale corn farming may be a household's only source of food in developing nations like Ethiopia. The aforementioned data demonstrates that the country's requirement for this crop is excessively high, and conversely, the crop's productivity is very low for a variety of reasons. The most damaging disease that greatly contributes to this imbalance between the crop's supply and demand is the corn disease. The failure to diagnose diseases in maize plant until they are too late is one of the most important factors influencing crop output in Ethiopia. This study will aid in the early detection of such diseases and support farmers during the cultivation process, directly affecting the amount of maize produced. The diseases in maize plants, such as northern leaf blight and cercospora leaf spot, have distinct symptoms that are visible. This study aims to detect the most frequent and degrading maize diseases using the most efficiently used subset of machine learning technology, deep learning so, called Image Processing. Deep learning uses networks that can be trained from unlabeled data without supervision (unsupervised). It is a feature that simulates the exercises the human brain goes through when digesting data. Its applications include speech recognition, language translation, object classification, and decision-making. Convolutional Neural Network (CNN) for Image Processing, also known as convent, is a deep learning class that is widely used for image classification, image detection, face recognition, and other problems. it will also use this algorithm as the state-of-the-art for my research to detect maize diseases by photographing maize leaves using a mobile phone.

Keywords: CNN, zea mays subsp, leaf blight, cercospora leaf spot

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Authors: Marwa Brahim, Nicolas Brosse, Nadia Boussetta, Nabil Grimi, Eugene Vorobiev

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Rapeseed plant is an established product in France which is mainly dedicated to oil production. However, the economic potential of residues from this industry (rapeseed hulls, rapeseed cake, rapeseed straw etc.), has not been fully exploited. Currently, only low-grade applications are found in the market. As a consequence, it was deemed of interest to develop a technological platform aiming to convert rapeseed residues into value- added products. Specifically, a focus is given on the conversion of rapeseed straw into valuable molecules (e.g. lignin, glucose). Existing pretreatment technologies have many drawbacks mainly the production of sugar degradation products that limit the effectiveness of saccharification and fermentation steps in the overall scheme of the lignocellulosic biorefinery. In addition, the viability of fractionation strategies is a challenge in an environmental context increasingly standardized. Hence, the need to find cleaner alternatives with comparable efficiency by implementing physical phenomena that could destabilize the structural integrity of biomass without necessarily using chemical solvents. To meet environmental standards increasingly stringent, the present work aims to study the new pretreatment strategies involving lower consumption of chemicals with an attenuation of the severity of the treatment. These strategies consist on coupling physical treatments either high voltage electrical discharges or ultrasounds to conventional chemical pretreatments (soda and organosolv). Ultrasounds treatment is based on the cavitation phenomenon, and high voltage electrical discharges cause an electrical breakdown accompanied by many secondary phenomena. The choice of process was based on a technological feasibility study taking into account the economic profitability of the whole chain after products valorization. Priority was given to sugars valorization into bioethanol and lignin sale.

Keywords: high voltage electrical discharges, organosolv, pretreatment strategies, rapeseed straw, soda, ultrasounds

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Authors: Peiming Li

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This paper presents an innovative blockchain-based approach to enhance the reliability and efficiency of federated learning systems. By integrating a decentralized oracle consensus mechanism into the federated learning framework, we address key challenges of data and model integrity. Our approach utilizes a network of redundant oracles, functioning as independent validators within an epoch-based training system in the federated learning model. In federated learning, data is decentralized, residing on various participants' devices. This scenario often leads to concerns about data integrity and model quality. Our solution employs blockchain technology to establish a transparent and tamper-proof environment, ensuring secure data sharing and aggregation. The decentralized oracles, a concept borrowed from blockchain systems, act as unbiased validators. They assess the contributions of each participant using a Hidden Markov Model (HMM), which is crucial for evaluating the consistency of participant inputs and safeguarding against model poisoning and malicious activities. Our methodology's distinct feature is its epoch-based training. An epoch here refers to a specific training phase where data is updated and assessed for quality and relevance. The redundant oracles work in concert to validate data updates during these epochs, enhancing the system's resilience to security threats and data corruption. The effectiveness of this system was tested using the Mnist dataset, a standard in machine learning for benchmarking. Results demonstrate that our blockchain-oriented federated learning approach significantly boosts system resilience, addressing the common challenges of federated environments. This paper aims to make these advanced concepts accessible, even to those with a limited background in blockchain or federated learning. We provide a foundational understanding of how blockchain technology can revolutionize data integrity in decentralized systems and explain the role of oracles in maintaining model accuracy and reliability.

Keywords: federated learning system, block chain, decentralized oracles, hidden markov model

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Authors: Muncharee Phaobthip

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In this research, the researcher re-examine the mediating effect of employee engagement between its antecedents and consequences for investigates the relation of leadership practices, employment branding and employee engagement based on social exchange theory. As such the researcher has four objectives as follows: First, to study the effects of leadership practices on employment branding, employee engagement and work intention; second, to examine the effects of employer brand perception on employee engagement and work intention; third, to examine the effects of employee engagement on work intention; and last, forth, the researcher inquires into the respondence of work intention. The researcher constituted a sample population of 535 employees of a Thai hotel chain located in four regions of the Kingdom of Thailand (Thailand). The researcher utilized a mixed-methods approach divided into quantitative and qualitative research investigatory phases, respectively. In the quantitative phase of research investigation, the researcher collected germane data from the 535 members of the sample population through the use of a questionnaire as a research instrument. In the qualitative phase of research investigation, relevant data were obtained through carrying out in-depth interviews with three subgroups of members of the sample population. These three subgroups consisted of twelve hotelier experts, six employees at the administrator level, and operational level employees. Focus group discussions were held with discussants from these three subgroups. Findings are as follows: Leadership practices showed positive effects on employment branding, employee engagement, and work intention. Employment branding displayed positive effects on employee engagement and work intention. Employee engagement had positive effects on work intention. However, in the analysis of the equation, the researcher confirmed that the important role of employee engagement is mediator factor between its antecedent and consequence factors. This provides benefits, in that it augments the body of knowledge devoted to the fostering of employee engagement in respect to psychological conditions. In conclusion, the researcher found that the value co-creation between leaders, employers and employees had positive effects on employee outcomes for lead to business outcomes according to reciprocal rule.

Keywords: antecedents, employee engagement, psychological conditions, work intention

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Authors: A. Kok

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The South African experience of the general upsurge in protest movements internationally is characterised by a tension between a neo-patrimonial state on the one hand, and a society with growing middle-class needs and interests on the other. This tension translates into local community service delivery protests – often violent in nature – that have been steadily increasing in number since 2008, student uprisings that have reached their height in October 2015, and various continuing local social #MustFall movements that are geared towards addressing government corruption and transforming neo-liberal structures. As a result, growing citizen (and non-citizen) revolt in South Africa has seen the (i) creeping securitization of the neo-patrimonial state and (ii) the 'top-down' misuse of a current 'bottom-up' people’s ideology, decoloniality, in an attempt by a faction in the ruling party (representing the neo-patrimonial state) to legitimize its actions and consolidate its power. The neo-patrimonial state’s creeping securitization and ideological positioning lead to a further mistrust of public institutions, people’s disengagement with traditional politics, and the creation of a 'parallel polis' by citizens and non-citizens that bypasses the official and oftentimes corrupt structures of the state. By applying the concept 'parallel polis' – originally developed by Václav Benda in connection with the movement Charter 77 in former Czechoslovakia – to a South African case study, it is illustrated that, even in the absence of overt oppression and the use of terror by a ruling elite, entrenched neo-patrimonialism can be potent enough to fuel the creation of various independent parallel public spheres (or, as a whole, understood as a 'parallel polis') to bypass dysfunctional state channels. A flourishing parallel polis offers possibilities for political, social and economic renewal. This is especially relevant in the consolidation of South Africa’s relatively young democracy.

Keywords: decoloniality, neo-patrimonialism, 'parallel polis', protest movements, South Africa, state securitization

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Authors: Fariha Akhter Chowdhury, Sabrina Mahboob, Anamika Saha, Afrin Jahan, Mohammad Nurul Islam

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Urinary tract infection (UTI) is frequently experienced by the female population where the prevalence increases with aging. Escherichia coli, one of the most common UTI causing organisms, retains glutathione defense mechanism that aids the organism to withstand the harsh physiological environment of urinary tract, host oxidative immune response and even to affect antibiotic-mediated cell death and the emergence of resistance. In this study, we aimed to investigate the glutathione reductase activity of uropathogenic E. coli (UPEC) by observing the reduced glutathione (GSH) level alteration under stressful condition. Urine samples of 58 patients with UTI were collected. Upon isolation and identification, 88% of the samples presented E. coli as UTI causing organism among which randomly selected isolates (n=9), obtained from urine samples of female patients, were considered for this study. E. coli isolates were grown under normal and stressful conditions where H₂O₂ was used as the stress-inducing agent. GSH level estimation of the isolates in both conditions was carried out based on the colorimetric measurement of 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) and GSH reaction product using microplate reader assay. The GSH level of isolated E. coli sampled from adult patients decreased under stress compared to normal condition (p = 0.011). On the other hand, GSH production increased markedly in samples that were collected from elderly subjects (p = 0.024). A significant partial correlation between age and change of GSH level was found as well (p = 0.007). This study may help to reveal ways for better understanding of E. coli pathogenesis of UTI prevalence in elderly patients.

Keywords: Escherichia coli, glutathione reductase activity, oxidative stress, reduced glutathione (GSH), urinary tract infection (UTI)

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