Search results for: Penosogan Formation

Authors: A. Guellati, F-M Lounis, N. Guemras, K. Daoud

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Micro droplet formation is considered as a growing emerging area of research due to its wide-range application in chemistry as well as biology. The mechanism of micro droplet formation using two immiscible liquids running through a T-junction has been widely studied. We believe that the flow of these two immiscible phases can be of greater important factor that could have an impact on out-flow hydrodynamic behavior, the droplets generated and the size of the droplets. In this study, the type of the capillary tubes used also represents another important factor that can have an impact on the generation of micro droplets. The tygon capillary tubing with hydrophilic inner surface doesn't allow regular out-flows due to the fact that the continuous phase doesn't adhere to the wall of the capillary inner surface. Teflon capillary tubing, presents better wettability than tygon tubing, and allows to obtain steady and regular regimes of out-flow, and the micro droplets are homogeneoussize. The size of the droplets is directly dependent on the flows of the continuous and dispersed phases. Thus, as increasing the flow of the continuous phase, to flow of the dispersed phase stationary, the size of the drops decreases. Inversely, while increasing the flow of the dispersed phase, to flow of the continuous phase stationary, the size of the droplet increases.

Keywords: microfluidic system, micro droplets generation, t-junction, fluids engineering

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Authors: Elakneswaran Yogarajah, Toyoharu Nawa, Eiji Owaki

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Cement-based materials have been using in various reinforced concrete structural components as well as in nuclear waste repositories. The sulfate attack has been an environmental issue for cement-based materials exposed to sulfate bearing groundwater or soils, and it plays an important role in the durability of concrete structures. The reaction between penetrating sulfate ions and cement hydrates can result in swelling, spalling and cracking of cement matrix in concrete. These processes induce a reduction of mechanical properties and a decrease of service life of an affected structure. It has been identified that the precipitation of secondary sulfate bearing phases such as ettringite, gypsum, and thaumasite can cause the damage. Furthermore, crystallization of soluble salts such as sodium sulfate crystals induces degradation due to formation and phase changes. Crystallization of mirabilite (Na₂SO₄:10H₂O) and thenardite (Na₂SO₄) or their phase changes (mirabilite to thenardite or vice versa) due to temperature or sodium sulfate concentration do not involve any chemical interaction with cement hydrates. Over the past couple of decades, an intensive work has been carried out on sulfate attack in cement-based materials. However, there are several uncertainties still exist regarding the mechanism for the damage of concrete in sulfate environments. In this study, modelling work has been conducted to investigate the chemical degradation of cementitious materials in various sulfate environments. Both internal and external sulfate attack are considered for the simulation. In the internal sulfate attack, hydrate assemblage and pore solution chemistry of co-hydrating Portland cement (PC) and slag mixing with sodium sulfate solution are calculated to determine the degradation of the PC and slag-blended cementitious materials. Pitzer interactions coefficients were used to calculate the activity coefficients of solution chemistry at high ionic strength. The deterioration mechanism of co-hydrating cementitious materials with 25% of Na₂SO₄ by weight is the formation of mirabilite crystals and ettringite. Their formation strongly depends on sodium sulfate concentration and temperature. For the external sulfate attack, the deterioration of various types of cementitious materials under external sulfate ingress is simulated through reactive transport model. The reactive transport model is verified with experimental data in terms of phase assemblage of various cementitious materials with spatial distribution for different sulfate solution. Finally, the reactive transport model is used to predict the long-term performance of cementitious materials exposed to 10% of Na₂SO₄ for 1000 years. The dissolution of cement hydrates and secondary formation of sulfate-bearing products mainly ettringite are the dominant degradation mechanisms, but not the sodium sulfate crystallization.

Keywords: thermodynamic calculations, reactive transport, radioactive waste disposal, PHREEQC

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Authors: Hao Liu, Lihui Zheng, Chinedu J. Okere, Chao Wang, Xiangchun Wang, Peng Zhang

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The coalbed methane (CBM) extraction industry (an unconventional energy source) is yet to promulgated an established standard code of practice for the experimental evaluation of sensitivity damage of coal samples. The existing experimental process of previous researches mainly followed the industry standard for conventional oil and gas reservoirs (CIS). However, the existing evaluation method ignores certain critical differences between CBM reservoirs and conventional reservoirs, which could inevitably result in an inaccurate evaluation of sensitivity damage and, eventually, poor decisions regarding the formulation of formation damage prevention measures. In this study, we propose improved experimental guidelines for evaluating seepage sensitivity damage of CBM reservoirs by leveraging on the shortcomings of the existing methods. The proposed method was established via a theoretical analysis of the main drawbacks of the existing methods and validated through comparative experiments. The results show that the proposed evaluation technique provided reliable experimental results that can better reflect actual reservoir conditions and correctly guide future development of CBM reservoirs. This study is pioneering the research on the optimization of experimental parameters for efficient exploration and development of CBM reservoirs.

Keywords: coalbed methane, formation damage, permeability, unconventional energy source

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Authors: Mohamad Saab, Florent Real, Francois Virot, Laurent Cantrel, Valerie Vallet

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All spent nuclear fuel reprocessing plants use the PUREX process (Plutonium Uranium Refining by Extraction), which is a liquid-liquid extraction method. The organic extracting solvent is a mixture of tri-n-butyl phosphate (TBP) and hydrocarbon solvent such as hydrogenated tetra-propylene (TPH). By chemical complexation, uranium and plutonium (from spent fuel dissolved in nitric acid solution), are separated from fission products and minor actinides. During a normal extraction operation, uranium is extracted in the organic phase as the UO₂(NO₃)₂(TBP)₂ complex. The TBP solvent can form an explosive mixture called red oil when it comes in contact with nitric acid. The formation of this unstable organic phase originates from the reaction between TBP and its degradation products on the one hand, and nitric acid, its derivatives and heavy metal nitrate complexes on the other hand. The decomposition of the red oil can lead to violent explosive thermal runaway. These hazards are at the origin of several accidents such as the two in the United States in 1953 and 1975 (Savannah River) and, more recently, the one in Russia in 1993 (Tomsk). This raises the question of the exothermicity of reactions that involve TBP and all other degradation products, and calls for a better knowledge of the underlying chemical phenomena. A simulation tool (Alambic) is currently being developed at IRSN that integrates thermal and kinetic functions related to the deterioration of uranyl nitrates in organic and aqueous phases, but not of the n-butyl phosphate. To include them in the modeling scheme, there is an urgent need to obtain the thermodynamic and kinetic functions governing the deterioration processes in liquid phase. However, little is known about the thermodynamic properties, like standard enthalpies of formation, of the n-butyl phosphate molecules and of the UO₂(NO₃)₂(TBP)₂ UO₂(NO₃)₂(HDBP)(TBP) and UO₂(NO₃)₂(HDBP)₂ complexes. In this work, we propose to estimate the thermodynamic properties with Quantum Methods (QM). Thus, in the first part of our project, we focused on the mono, di, and tri-butyl complexes. Quantum chemical calculations have been performed to study several reactions leading to the formation of mono-(H₂MBP), di-(HDBP), and TBP in gas and liquid phases. In the gas phase, the optimal structures of all species were optimized using the B3LYP density functional. Triple-ζ def2-TZVP basis sets were used for all atoms. All geometries were optimized in the gas-phase, and the corresponding harmonic frequencies were used without scaling to compute the vibrational partition functions at 298.15 K and 0.1 Mpa. Accurate single point energies were calculated using the efficient localized LCCSD(T) method to the complete basis set limit. Whenever species in the liquid phase are considered, solvent effects are included with the COSMO-RS continuum model. The standard enthalpies of formation of TBP, HDBP, and H2MBP are finally predicted with an uncertainty of about 15 kJ mol⁻¹. In the second part of this project, we have investigated the fundamental properties of three organic species that mostly contribute to the thermal runaway: UO₂(NO₃)₂(TBP)₂, UO₂(NO₃)₂(HDBP)(TBP), and UO₂(NO₃)₂(HDBP)₂ using the same quantum chemical methods that were used for TBP and its derivatives in both the gas and the liquid phase. We will discuss the structures and thermodynamic properties of all these species.

Keywords: PUREX process, red oils, quantum chemical methods, hydrolysis

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Authors: Alono Thorie, Ananya Mukhopadhyay

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Ooid bearing horizons of the Proterozoic Kunihar Formation, Simla Group, Lesser Himalaya have been addressed in the present work. The study is concentrated around the outskirts of Arki town, Solan district, Himachal Pradesh, India. Based on the sedimentary facies associations, the processes that promote the formation of ooids have been documented. The facies associations that have been recorded are: (i) Oolitic-Intraclastic grainstone (FA1), (ii) Oolitic grainstone (FA2), (iii) Boundstone (FA3), (iv) Dolomudstone (FA4) and (v) Rudstone (FA5). Oolitic-Intraclastic grainstone (FA1) mainly consists of well sorted ooids with concentric laminae and intraclasts. Large ooids with grain sizes more than 4 mm are characteristic of oolites throughout the area. Normally graded beds consisting of ooids and intraclasts are frequently documented in storm sediments in shelf environments and carbonate platforms. The well-sorted grainstone fabric indicates deposition in a high-energy shoal with tidal currents and storm reworking. FA2 comprises spherical to elliptical grains up to 8.5cm in size with concentric cortex and micritic nuclei. Peloids in FA2 are elliptical, rounded objects <0.3 mm in size. FA1 and FA2 have been recorded alongside boundstones (FA3) comprising stromatolites having columnar, wavy and domal morphology. Boundstones (FA3) reflect microbial growth in carbonate platforms and reefs. Dolomudstones (FA4) interbedded with cross laminated sandstones and erosional surfaces reflect sedimentation in storm dominated zones below fair-weather wave base. Rudstone (FA5) is composed of oolitic grainstone (FA2), boundstone (FA3) and dolomudstone (FA4). These clasts are few mm to more than 10 cm in length. Rudstones indicate deposition along a slope with intermittent influence of wave currents and storm activities. Most ooids from the Kunihar Formation are regular ooids with abundance of broken ooids. Compound and concentric ooids indicating medium to low energy environments are present but scarce. Ooids from high energy domains are more dominant than ooids developed from low energy environments. The unusually large size of the Kunihar ooids (more than 8.5 cm) is rare in the geological record. Development of carbonate deposits such as oolitic- intraclastic Grainstones (FA1), oolitic grainstones (FA2) and rudstones (FA5), and reflect deposition in an agitated beach environment with abundant microbial activity and high energy shallow marine waters influenced by tide, wave and storm currents. Occurrences of boundstone (FA4) or stromatolitic carbonate amongst oolitic facies (FA1 and FA2) and appearance of compound and concentric ooids indicate intervals of calm in between agitated phases of storm, wave and tidal activities.

Keywords: proterozoic, Simla Group, ooids, stromatolites

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Authors: Temitope L. Baiyegunhi, Kuiwu Liu, Oswald Gwavava, Christopher Baiyegunhi

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Shale gas has recently been the exploration focus for future energy resource in South Africa. Specifically, the black shales of the lower Ecca Group in the study area are considered to be one of the most prospective targets for shale gas exploration. Evaluation of this potential resource has been restricted due to the lack of exploration and scarcity of existing drill core data. Thus, only limited previous geochemical data exist for these formations. In this study, outcrop and core samples of the Ecca Group were analysed to assess their total organic carbon (TOC), organic matter type, thermal maturity and hydrocarbon generation potential (SP). The results show that these rocks have TOC ranging from 0.11 to 7.35 wt.%. The SP values vary from 0.09 to 0.53 mg HC/g, suggesting poor hydrocarbon generative potential. The plot of S1 versus TOC shows that the source rocks were characterized by autochthonous hydrocarbons. S2/S3 values range between 0.40 and 7.5, indicating Type- II/III, III, and IV kerogen. With the exception of one sample from the collingham formation which has HI value of 53 mg HC/g TOC, all other samples have HI values of less than 50 mg HC/g TOC, thus suggesting Type-IV kerogen, which is mostly derived from reworked organic matter (mainly dead carbon) with little or no potential for hydrocarbon generation. Tmax values range from 318 to 601℃, indicating immature to over-maturity of hydrocarbon. The vitrinite reflectance values range from 2.22 to 3.93%, indicating over-maturity of the kerogen. Binary plots of HI against OI and HI versus Tmax show that the shales are of Type II and mixed Type II-III kerogen, which are capable of generating both natural gas and minor oil at suitable burial depth. Based on the geochemical data, it can be inferred that the source rocks are immature to over-matured variable from localities and have potential of producing wet to dry gas at present-stage. Generally, the Whitehill formation of the Ecca Group is comparable to the Marcellus and Barnett Shales. This further supports the assumption that the Whitehill Formation has a high probability of being a profitable shale gas play, but only when explored in dolerite-free area and away from the Cape Fold Belt.

Keywords: source rock, organic matter type, thermal maturity, hydrocarbon generation potential, Ecca Group

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Authors: Babatunde Oluwole Ogunsile, Omosola Monisola Fasoranti

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High costs and toxicological hazards associated with the physicochemical methods of producing nanoparticles have limited their widespread use in clinical and biomedical applications. An ethically sound alternative is the utilization of plant bioresources as a low cost and eco–friendly biological approach. Silver nanoparticles (AgNPs) were synthesized from aqueous leaf extract of Tithonia diversifolia plant. The UV-Vis Spectrophotometer was used to monitor the formation of the AgNPs at different time intervals and different ratios of plant extract to the AgNO₃ solution. The biosynthesized AgNPs were characterized by FTIR, X-ray Diffraction (XRD) and Scanning Electron Microscope (SEM). Antimicrobial activities of the AgNPs were investigated against ten human pathogens using agar well diffusion method. The AgNPs yields were modeled using a second-order factorial design. The result showed that the rate of formation of the AgNPs increased with respect to time while the optimum ratio of plant extract to the AgNO₃ solution was 1:1. The hydroxyl group was strongly involved in the bioreduction of the silver salt as indicated by the FTIR spectra. The synthesized AgNPs were crystalline in nature, with a uniformly distributed network of the web-like structure. The factorial model predicted the nanoparticles yields with minimal errors. The nanoparticles were active against all the tested pathogens and thus have great potentials as antimicrobial agents.

Keywords: antimicrobial activities, green synthesis, silver nanoparticles, Tithonia diversifolia

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Authors: Maryam Pirdehghan

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The myth of Leviathan, its ontology and appearance is often one of the problems of Judeo-Christian religious commentators so that some of them have tried to interpret and explain formation or symbolic implications of this myth in different contexts their specific methods and proofs. However, the Bible has presented only vague references in this field and it is not clear why and how to develop such mentions to create a powerful myth with allegorical and symbolic capacity as Leviathan. Therefore, the paper aims to clarify the process of formation of Leviathan and explore the mythical and symbolic systems related to it, first by adopting the imagological approach and then using the Northrop Frye's Archetypal Criticism. Finally, it is concluded that The Leviathan is rooted in the stories of legendary battles of the beginning of creation and almost continues to live with the same nature into the Old Testament, but continuously, in an interactive process between the Greek and Egyptian mythological networks, it attracts more stories and implications about his existence while maintaining its satanic nature. After intense metamorphosis in Jewish interpretations, it appears in the book of Revelation and finally, becomes one of the princes of Hell in the tradition of Christian demonology. The myth, that has become the archetype and fluidized symbol of evil because of the ambiguity and lack of objectivity on its apparent characteristics, finds symbolical extensive capabilities in Judeo-Christian culture, especially in the mysticism, so that its presence or death has special implications and also fighting against it is taken into account as an external and more internal action.

Keywords: Leviathan, The Evil, Bible, myth, Northrop Frye

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Authors: Christopher Viens, Steve Krase

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Terms such as “riding a hard streak”, “formation push”, and “fighting formation” are commonly used in the directional drilling world to explain BHA behavior that causes unwanted trajectory change. Theories about downhole directional tendencies are commonly speculated from various personal experiences with little merit due to the lack of hard data to reveal the actual mechanisms behind the phenomenon, leaving interpretation of the root cause up to personal perception. Understanding and identifying in real time the lithological factors that influence the BHA to change or hold direction adds tremendous value in terms reducing sliding time and targeting zones for optimal ROP. Utilizing surface drilling parameters and employing downhole measurements of azimuthal gamma, continuous inclination, and bending moment, a direct measure of the rock related directional phenomenon have been captured and quantified. Furthermore, identifying continuous zones of like lithology with consistent bit to rock interaction has value from a reservoir characterization and completions standpoint. The paper will show specific examples of lithology related directional tendencies from the Spraberry and Wolfcamp in the Delaware Basin.

Keywords: Azimuthal gamma imaging, bending moment, continuous inclination, downhole dynamics measurements, high frequency data

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Authors: Michal Natan, Ori Gutman, Shlomo Margel, Ehud Banin

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Biofilm formation is a serious problem in medical and industrial settings due to the increased resistance of these communities to killing compared to free-living bacteria. This has prompted the search for agents that can inhibit both bacterial growth and biofilm formation. In this study, N-halamine rechargeable nanoparticles (NPs) were synthesized by co-polymerization of the monomer methacryl amide and the cross-linker monomer N,N-methylene bisacryl amide, and were subsequently loaded with Cl+, using bleach. The chlorinated NPs exhibited remarkable stability to organic reagents. The antibacterial mechanism of the P(MAA-MBAA)-Cl NPs involved generation of reactive oxygen species (ROS) only upon exposure to organic media, but not upon incubation in water, suggesting a specific activation. Moreover, a unique interaction of the P(MAA-MBAA)-Cl NPs with Staphylococcus aureus bacteria but not with human cells was discovered, whereby these microorganisms were all specifically targeted and marked for destruction. Finally, in collaboration with Netafim Ltd. irrigation drippers containing the P(MAA-MBAA)-Cl were incubated in the field and were shown to prevent fouling on them for 5 months as opposed to the control drippers that exhibited substantial fouling. Further, the NPs offer recharging to the surface, thus providing long-lasting protection that does not exist in the products available today. Taken together, the results demonstrate the great potential of implementing the charged NPs in devices and surfaces to prevent bacterial growth.

Keywords: bacteria, biofilm, fouling, nanoparticles

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Authors: M. Vadivel, R. Ramesh Babu

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Spinel ferrite magnetic nanomaterials have received a great deal of attention in recent years due to their wide range of potential applications in various fields such as magnetic data storage and microwave device applications. Among the family of spinel ferrites, cobalt ferrite (CoFe2O4) has been widely used in the field of high-frequency applications because of its remarkable material qualities such as moderate saturation magnetization, high coercivity, large permeability at higher frequency and high electrical resistivity. For aforementioned applications, the materials should have an improved electrical property, especially enhancement in the dielectric properties. It is well known that the substitution of rare earth metal cations in Fe3+ site of CoFe2O4 nanoparticles leads to structural distortion and thus significantly influences the structural and morphological properties whereas greatly modifies the electrical and magnetic properties of a material. In the present investigation, we report on the influence of lanthanum (La3+) ion substitution on the structural, morphological, dielectric and magnetic properties of CoFe2O4 magnetic nanoparticles prepared by co-precipitation method. Powder X-ray diffraction patterns reveal the formation of inverse cubic spinel structure with the signature of LaFeO3 phase at higher La3+ ion concentrations. Raman and Fourier transform infrared spectral analysis also confirms the formation of inverse cubic spinel structure and Fe-O symmetrical stretching vibrations of CoFe2O4 nanoparticles, respectively. Transmission electron microscopy study reveals that the size of the particles gradually increases with increasing La3+ ion concentrations whereas the agglomeration gets slightly reduced for La3+ ion substituted CoFe2O4 nanoparticles than that of undoped CoFe2O4 nanoparticles. Dielectric properties such as dielectric constant and dielectric loss were recorded as a function of frequency and temperature which reveals that the dielectric constant gradually increases with increasing temperatures as well as La3+ ion concentrations. The increased dielectric constant might be the reason that the formation of LaFeO3 secondary phase at higher La3+ ion concentrations. Magnetic measurement demonstrates that the saturation magnetization gradually decreases from 61.45 to 25.13 emu/g with increasing La3+ ion concentrations which is due to the nonmagnetic nature of La3+ ions substitution.

Keywords: cobalt ferrite, co-precipitation, dielectric properties, saturation magnetization

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Authors: Pouyan Motamedi, Ken Bosnick, Ken Cadien, James Hogan

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Growth of conformal ultrathin metal films has attracted a considerable amount of attention recently. Plasma-enhanced atomic layer deposition (PEALD) is a method capable of growing conformal thin films at low temperatures, with an exemplary control over thickness. The authors have recently reported on growth of metastable epitaxial nickel thin films via PEALD, along with a comprehensive characterization of the films and a study on the relationship between the growth parameters and the film characteristics. The goal of the current study is to use the mentioned films as a case study to investigate the temperature-activated phase transformation and agglomeration in ultrathin metallic films. For this purpose, metastable hexagonal nickel thin films were annealed using a controlled heating/cooling apparatus. The transformations in the crystal structure were observed via in-situ synchrotron x-ray diffraction. The samples were annealed to various temperatures in the range of 400-1100° C. The onset and progression of particle formation were studied in-situ via laser measurements. In addition, a four-point probe measurement tool was used to record the changes in the resistivity of the films, which is affected by phase transformation, as well as roughening and agglomeration. Thin films annealed at various temperature steps were then studied via atomic force microscopy, scanning electron microscopy and high-resolution transmission electron microscopy, in order to get a better understanding of the correlated mechanisms, through which phase transformation and particle formation occur. The results indicate that the onset of hcp-to-bcc transformation is at 400°C, while particle formations commences at 590° C. If the annealed films are quenched after transformation, but prior to agglomeration, they show a noticeable drop in resistivity. This can be attributed to the fact that the hcp films are grown epitaxially, and are under severe tensile strain, and annealing leads to relaxation of the mismatch strain. In general, the results shed light on the nature of structural transformation in nickel thin films, as well as metallic thin films, in general.

Keywords: atomic layer deposition, metastable, nickel, phase transformation, thin film

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Authors: Mahmood Seyyed, Akram Salehi Heykoei, Hamidreza Safakish Kashani

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Among the remaining resource of the past, coins considered as an authentic documents among the most important documentary sources. The coins were minted in a place that called mint. The number and position of the mints in each period reflects the amount of economic power, political security and business growth, which was always fluctuated its position with changing the political and economic condition. Considering that, trade has more growth during the Safavid period than previous ones, the mint also has been in greater importance. It seems the one hand, the growth of economic in Safavid period has a direct link with the number and places of the mints at that time and in the other hand, the mints have been formed in some places because of the specific characteristic of cities and regions. Increasing the number of mints in the north of the country due to the growth of silk trade and in the west and northwest due to the political and commercial relation with Ottoman Empire, also the characteristics such as existence of mines, located in the Silk Road and communication ways, all are the results of this investigation. Accordingly, in this article researcher tries to examine the characteristics that give priority to a city for having mint. With considering that in the various historical periods, the mints were based in the most important cities in terms of political and social, at that time, this article examines the cities specific characteristics in the formation of the mints in Safavid period.

Keywords: documentary sources, coins, mint, city, Safavid

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Authors: Abilio Avila, Orestis Terzidis

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A fundamental principle of Grounded Theory (GT) is to prevent the formation of preconceived theories. This implies the need to start a research study with an open mind and to avoid being absorbed by the existing literature. However, to start a new study without an understanding of the research domain and its context can be extremely challenging. This paper presents a research approach that simultaneously supports a researcher to identify and to focus on critical areas of a research project and prevent the formation of prejudiced concepts by the current body of literature. This approach comprises of four stages: Selection of secondary case studies, analysis of secondary case studies, development of an initial conceptual framework, development of an initial interview guide. The analysis of secondary case studies as a starting point for a research project allows a researcher to create a first understanding of a research area based on real-world cases without being influenced by the existing body of theory. It enables a researcher to develop through a structured course of actions a firm guide that establishes a solid starting point for further investigations. Thus, the described approach may have significant implications for GT researchers who aim to start a study within a given research area.

Keywords: grounded theory, interview guide, qualitative research, secondary case studies, secondary data analysis

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Authors: Amokrane Boufares, Elise Provost, Veronique Osswald, Pascal Clain, Anthony Delahaye, Laurence Fournaison, Didier Dalmazzone

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Gas hydrates have long been considered as problematic for flow assurance in natural gas and oil transportation. On the other hand, they are now seen as future promising materials for various applications (i.e. desalination of seawater, natural gas and hydrogen storage, gas sequestration, gas combustion separation and cold storage and transport). Nonetheless, a better understanding of the crystallization mechanism of gas hydrate and of their formation kinetics is still needed for a better comprehension and control of the process. To that purpose, measuring the real-time evolution of the dissolved gas concentration in the aqueous phase during hydrate formation is required. In this work, CO₂ hydrates were formed in a stirred reactor equipped with an Attenuated Total Reflection (ATR) probe coupled to a Fourier Transform InfraRed (FTIR) spectroscopy analyzer. A method was first developed to continuously measure in-situ the CO₂ concentration in the liquid phase during solubilization, supersaturation, hydrate crystallization and dissociation steps. Thereafter, the measured concentration data were compared with those of equilibrium concentrations. It was observed that the equilibrium is instantly reached in the liquid phase due to the fast consumption of dissolved gas by the hydrate crystallization. Consequently, it was shown that hydrate crystallization kinetics is limited by the gas transfer at the gas-liquid interface. Finally, we noticed that the liquid-hydrate equilibrium during the hydrate crystallization is governed by the temperature of the experiment under the tested conditions.

Keywords: gas hydrate, dissolved gas, crystallization, infrared spectroscopy

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Authors: S. Teimouri, S. Abbasi

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Milk-fruit juice mixture is a type of soft drinks, which can be produced by mixing milk with pieces of fruits, fruit juices, or fruit juices concentrates. The major problem of these products, mainly the acidic ones, is phase separation which occurs during formulation and storage due to the aggregation of caseins at low pH Short-chain inulin (CLR), long-chain inulin (TEX), native inulin (IQ) and Long-chain inulin (TEX) and short-chain inulin (CLR) combined in different proportions (2o:80, 50:50, and 80:20) were added (2-10 %) to sourcherry juice-milk mixture and their stabilization mechanisms were studied with using rheological and microstructural observations. Stevioside as a bio-sweetener and sugar-replacer was added at last step. Finally, sensory analyses were taken place on stabilized samples. According to the findings, TEX stabilized the mixture at concentration of 8%. MIX and IQ reduced phase separation at high concentration but had not complete effect on stabilization. CLR did not effect on stabilization. Rheological changes and inulin aggregates formation were not observed in CLR samples during the one month storage period. However TEX, MIX and IQ samples formed inulin aggregates and became more thixotropic, elastic and increased the viscosity of mixture. The rate of the inulin aggregates formation and viscosity increasing was in the following order TEX > MIX > IQ. Consequently the mixture which stabilized with inulin and sweetened with stevioside had the prebiotic properties which may suggest to diabetic patients and children.

Keywords: prebiotic, inulin, casein, stabilization, stevioside

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Authors: Richard Lewis

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The following assumptions of the Big Bang theory are challenged and found to be false: the cosmological principle, the assumption that all matter formed at the same time and the assumption regarding the cause of the cosmic microwave background radiation. The evolution of the universe is described based on the conclusion that the universe is finite with a space boundary. This conclusion is reached by ruling out the possibility of an infinite universe or a universe which is finite with no boundary. In a finite universe, the centre of the universe can be located with reference to our home galaxy (The Milky Way) using the speed relative to the Cosmic Microwave Background (CMB) rest frame and Hubble's law. This places our home galaxy at a distance of approximately 26 million light years from the centre of the universe. Because we are making observations from a point relatively close to the centre of the universe, the universe appears to be isotropic and homogeneous but this is not the case. The CMB is coming from a source located within the event horizon of the universe. There is sufficient mass in the universe to create an event horizon at the Schwarzschild radius. Galaxies form over time due to the energy released by the expansion of space. Conservation of energy must consider total energy which is mass (+ve) plus energy (+ve) plus spacetime curvature (-ve) so that the total energy of the universe is always zero. The predominant position of galaxy formation moves over time from the centre of the universe towards the boundary so that today the majority of new galaxy formation is taking place beyond our horizon of observation at 14 billion light years.

Keywords: cosmology, dark energy, dark matter, evolution of the universe

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Authors: Rui Tu, Yakui Bai, Huailin Li

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The lead-bismuth eutectic (LBE) alloy is a promising candidate of coolant in the fast neutron reactors and accelerator-driven systems (ADS) because of its good properties, such as low melting point, high neutron yields and high thermal conductivity. Although the corrosion of the structure materials caused by the liquid metal (LM) coolant is a challenge to the safe operating of a lead-bismuth eutectic nuclear reactor. Thermodynamic theories, experiential formulas and experimental data can be used for explaining the maintenance of the protective oxide layers on stainless steels under satisfaction oxygen concentration, but the atomic scale insights of such anti-corrosion mechanisms are little known. In the present work, the first-principles calculations are carried out to study the effects of oxygen partial pressure on the formation energies of the liquid metal coolant relevant impurity defects in the anti-corrosion oxide films on the surfaces of the structure materials. These approaches reveal the microscope mechanisms of the corrosion of the structure materials, especially for the influences from the oxygen partial pressure. The results are helpful for identifying a crucial oxygen concentration for corrosion control, which can ensure the systems to be operated safely under certain temperatures.

Keywords: oxygen partial pressure, liquid metal coolant, TDDFT, anti-corrosion layer, formation energy

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Authors: Ahmed Abdulrahman, Jalal Foroozesh

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CO₂ based enhanced oil recovery (EOR) techniques have gained massive attention from major oil firms since they resolve the industry's two main concerns of CO₂ contribution to the greenhouse effect and the declined oil production. Carbonated water injection (CWI) is a promising EOR technique that promotes safe and economic CO₂ storage; moreover, it mitigates the pitfalls of CO₂ injection, which include low sweep efficiency, early CO₂ breakthrough, and the risk of CO₂ leakage in fractured formations. One of the main challenges that hinder the wide adoption of this EOR technique is the complexity of accurate modeling of the kinetics of CO₂ mass transfer. The mechanisms of CO₂ mass transfer during CWI include the slow and gradual cross-phase CO₂ diffusion from carbonated water (CW) to the oil phase and the CO₂ dispersion (within phase diffusion and mechanical mixing), which affects the oil physical properties and the spatial spreading of CO₂ inside the reservoir. A 2D non-equilibrium compositional simulator has been developed using a fully implicit finite difference approximation. The material balance term (k) was added to the governing equation to account for the slow cross-phase diffusion of CO₂ from CW to the oil within the gird cell. Also, longitudinal and transverse dispersion coefficients have been added to account for CO₂ spatial distribution inside the oil phase. The CO₂-oil diffusion coefficient was calculated using the Sigmund correlation, while a scale-dependent dispersivity was used to calculate CO₂ mechanical mixing. It was found that the CO₂-oil diffusion mechanism has a minor impact on oil recovery, but it tends to increase the amount of CO₂ stored inside the formation and slightly alters the residual oil properties. On the other hand, the mechanical mixing mechanism has a huge impact on CO₂ spatial spreading (accurate prediction of CO₂ production) and the noticeable change in oil physical properties tends to increase the recovery factor. A sensitivity analysis has been done to investigate the effect of formation heterogeneity (porosity, permeability) and injection rate, it was found that the formation heterogeneity tends to increase CO₂ dispersion coefficients, and a low injection rate should be implemented during CWI.

Keywords: CO₂ mass transfer, carbonated water injection, CO₂ dispersion, CO₂ diffusion, cross phase CO₂ diffusion, within phase CO2 diffusion, CO₂ mechanical mixing, non-equilibrium simulation

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Authors: Abdul M. Khan, Hafiza I. Naz, Ayesha Iqbal, Muhammad Akhtar

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The fossil remains described in this study have been recovered during fieldwork by the authors from the Dhok Pathan Formation of Middle Siwaliks Pakistan in December, 2015. The sample comprises maxillary and mandibular fragments along with isolated upper and lower teeth. The morphometric analysis of the specimens led us to recognize the sample as belonging to Eotragus noyei, which has been considered as the smallest and the oldest bovid in the Siwaliks. Eotragus noyei is characterized by brachydont teeth, finely rugose enamel, more inclined buccal walls of the molars and small lingual cingula. The inclination of the metaconal area has caused rotation of the metastyle in relation to the antero-posterior tooth axis and thus situated more lingually. The protocone in second upper premolar is well developed and situated posteriorly and also has an anterior lingual constriction. The metaconule in the third upper molar is smaller than the protocone. The dentition in Eotragus noyei is smaller in size as compared to Eotragus sansaniensis and Eotragus lampangensis. In Eotragus noyei the buccal walls in molars are more inclined while in Eotragus sansaniensis they are less inclined. The genus Eotragus has been reported previously in the Lower and Middle Siwaliks of Pakistan; however, the recognition of the present sample as Eotragus noyei has extended the range of this species from Lower to the Middle Siwaliks of Pakistan.

Keywords: Boselaphini, Chakwal, Dhok Pathan, late miocene

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Authors: Anusmita Das

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In the post-industrial scenario, cities in India have become critical sites of negotiation and are expected to become some of the largest urban agglomeration of the twenty-first century. This has created a pluralist identity resulting in a new multifarious urbanism pervading throughout the entire urban landscape. There is an ambiguity regarding the character of present day Indian cities with new meanings emerging and no methodical study to understand them. More than an abstract diagram, the present day cities can be looked at as an ensemble of meanings. One of the ways in which the meaning is reflected is through events. Festivals such as Diwali, Dussera, Durga Puja, Ganesh Chaturthi, etc have transpired as the phenomenon of the city, and their presence in the everyday landscape weaves itself through the urban fabric dominating the popular visual culture of Indian cities. Festivals influence people’s idea of a city. Ritual, festival, celebrations are important in shaping of the urban environment and in their influence on the intangible aspect of the urban setting. These festivals pertaining to the city in motion have emerged as the symbolic image of the emerging urban Indian condition giving birth to new urban identities. The study undertaken to understand the present context of temporality of Indian cities is important in analyzing the process of its formation and transformation. This study aims to review the evolution of new dimensions of urbanism in India as well as its implication on the identity of cities.

Keywords: urban identities, urban design, festivals, rituals, celebrations, inter-disciplinary study

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Authors: Susanna Menis

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The context of this study revolves around the theme of consent and the construction of unlawfulness in judicial decisions. It aims to explore the formation of societal perceptions of unlawfulness within the context of consensual sexual acts leading to harmful consequences. This study investigates how judges create legal rules that reflect social solidarity and protect against violence. Specifically, the research aims to understand the justification behind criminalising consensual sexual activity when categorised under different offences. The main question addressed in this study will evaluate the way judges create legal rules that they believe reflect social solidarity and protect against violence. The study employs a historical genealogy approach as its methodology. This approach allows for tracing back the original formation of societal perspectives on unlawfulness, thus highlighting the socially constructed nature of the present understanding. The data for this study will be collected through an extensive literature review, examining historical legal cases and documents that shape the understanding of unlawfulness. This will provide a comprehensive view of how social attitudes toward private sexual relations influenced the creation of legal rules. The theoretical importance of this research lies in its contribution to socio-legal scholarship. This study adds to the existing knowledge on the topic by exploring questions of unconscious bias and its origins. The findings shed light on how and why individuals possess unconscious biases, particularly within the judicial system. In conclusion, this study investigates judicial decisions concerning consensual sexual acts and the construction of unlawfulness. By employing a historical genealogy approach, the research sheds light on how judges create legal rules that reflect social solidarity and aim to protect against violence. The theoretical importance of this study lies in its contribution to understanding unconscious bias and its origins within the judicial system. Through data collection and analysis procedures, this study aims to provide valuable insights into the formation of social attitudes towards private sexual relations and its impact on legal rulings.

Keywords: consent, sexual offences, offences against the person, legal genealogy, social construct

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Authors: Natalia Hordynska, Magdalena Szechynska-Hebda, Miroslaw Sobczak, Elzbieta Rozanska, Joanna Troczynska, Zofia Banaszak, Maria Wedzony

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Albinism is a common problem encountered in wheat and triticale breeding programs, which require in vitro culture steps e.g. generation of doubled haploids via androgenesis process. Genetic factor is a major determinant of albinism, however, environmental conditions such as temperature and media composition influence the frequency of albino plant formation. Cold incubation of wheat and triticale spikes induced a switch from gametophytic to sporophytic development. Further, androgenic structures formed from anthers of the genotypes susceptible to androgenesis or treated with cold stress, had a pool of structurally primitive plastids, with small starch granules or swollen thylakoids. High temperature was a factor inducing andro-genesis of wheat and triticale, but at the same time, it was a factor favoring the formation of albino plants. In genotypes susceptible to albinism or after heat stress conditions, cells formed from anthers were vacuolated, and plastids were eliminated. Partial or complete loss of chlorophyll pigments and incomplete differentiation of chloroplast membranes result in formation of tissues or whole plant unable to perform photosynthesis. Indeed, susceptibility to the andro-genesis process was associated with an increase of total concentration of photosynthetic pigments in anthers, spikes and regenerated plants. The proper balance of the synthesis of various pigments, was the starting point for their proper incorporation into photosynthetic membranes. In contrast, genotypes resistant to the androgenesis process and those treated with heat, contained 100 times lower content of photosynthetic pigments. In particular, the synthesis of violaxanthin, zeaxanthin, lutein and chlorophyll b was limited. Furthermore, deregulation of starch and lipids synthesis, which led to the formation of very complex starch granules and an increased number of oleosomes, respectively, correlated with the reduction of the efficiency of androgenesis. The content of other sugars varied depending on the genotype and the type of stress. The highest content of various sugars was found for genotypes susceptible to andro-genesis, and highly reduced for genotypes resistant to androgenesis. The most important sugars seem to be glucose and fructose. They are involved in sugar sensing and signaling pathways, which affect the expression of various genes and regulate plant development. Sucrose, on the other hand, seems to have minor effect at each stage of the androgenesis. The sugar metabolism was related to metabolic activity of microspores. The genotypes susceptible to androgenesis process had much faster mitochondrium- and chloroplast-dependent energy conversion and higher heat production by tissues. Thus, the effectiveness of metabolic processes, their balance and the flexibility under the stress was a factor determining the direction of microspore development, and in the later stages of the androgenesis process, a factor supporting the induction of androgenic structures, chloroplast formation and the regeneration of green plants. The work was financed by Ministry of Agriculture and Rural Development within Program: ‘Biological Progress in Plant Production’, project no HOR.hn.802.15.2018.

Keywords: androgenesis, chloroplast, metabolism, temperature stress

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Authors: Ravi Teja Chitturi Suryaprakash, Chandrashekhar Unakal, Haytham Al-Bayaty, Duraisamy Saravanakumar

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Background and significance: Oral biofilm formation is a well-known causative factor for caries and periodontal diseases. Scientists over the years have been trying to find a solution against the formation of oral biofilms. Though several advances have been made to understand the microbial ecology and how the bio film survives, it is still an enigma to researchers to find a chemical product that not only can inhibit the formation of oral bio film but also not disturb the oral micro flora required for oral health and not to cause damage to the cells of the oral cavity. One such product that has never been investigated much are herbal preparations. Some of the microorganisms important in the formation of biofilm are Streptococcus mutans, Actinomyces naeslundi, Streptococuss oralis and Prevotella intermedia. The aim of this study was to study the antimicrobial property of some herbal extracts available in Trinidad and Tobago against these pathogens. The significance of this study is that identification of biologically effective plant extracts can result in indigenous development of mouth rinses and tooth pastes that the people can benefit from to not only develop effective but also a cheap solution. Methodology: The extracts from the leaves of Plectranthus ambonicus, Ocmium tenuiflorum, Azadirchata indica, Anacardium occidentale, Psidium guajava were prepared by dissolving them in water. The extracts from the roots of Curcuma longa were prepared similarly and the antimicrobial activity of these six plant extracts was determined by the agar well diffusion method using minimum inhibitory concentration (MIC) against Streptococcus mutans, Actinomyces naeslundi, Streptococuss oralis and Prevotella intermedia and compared with chlorhexidine. Results: The six plant extracts showed variable effect on the oral micro-organisms. Ocmium tenuiflorum (16.66 ± 0.44, 14 ± 0.58, 13.33 ± 0.88, 12.83 ± 0.60), Azadirchata indica (17.5 ± 0.28, 14.83 ± 0.17, 15 ± 0.58, 12.83 ± 0.6) and Curcuma longa (16.16 ± 0.44, 13.66 ± 0.88, 12.33 ± 0.88, 11.33 ± 0.67) were found to have highest inhibitory activity against all the four pathogens (Streptococcus mutans, Streptococuss oralis, Actinomyces naeslundi, and Prevotella intermedia) respectively. Conclusion: Although the extracts were not pure compounds we obtained antimicrobial results which determine that they are potent antimicrobial agents. Further derivation of pure compounds from these extracts could be lucrative as it might lead to the development of a cost effective and biologically safe medicine to act against oral biofilms. Acknowledgement: The authors would like to acknowledge the Campus Research and Publication Fund Committee, The University of the West Indies for funding this study and would also like to acknowledge Dr. Leonette Cox, Department of Chemistry, Faculty of Science and Technology, The University of the West Indies, St. Augustine Campus, Trinidad and Tobago for helping to prepare the plant extracts.

Keywords: agar well diffusion method, herbal extracts, minimum inhibitory concentration, oral biofilm forming microorganisms

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Authors: Manh Tin Ho, Phorl Sophors, Ga Young Seo, Young Mee Kim, Youngho Lim, Moonjae Cho

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UV radiation in sunlight is one of the most potential factor induced skin ageing and photocarcinogenesis. UV may induce the melanin production and wrinkle formation. Recently, the natural secondary compounds have been reported that had the beneficial protective effects from UV light. In this study, we investigated the effects of two different compounds, glycitin and 38, on human dermal fibroblast. We first only treated the 38 on melanocyte cell to test the proliferation inhibition of 38 on this cell line. Then, we induced the combination of glycitin and 38 on human dermal fibroblast in 48h and investigate the proliferation, collagen production and the metalloproteinase family expression. The 38 alone could inhibit the proliferation of melanocyte which indicated the reduction of melanin production. The combination of glycitin and 38 truly increased the fibroblast proliferation and even they could recover the UV-induced and H2O2-induced damaged fibroblast proliferation. The co-treatment also promoted the collagen IV expression significantly and accelerated the total collagen secretion. In addition, metalloproteinase (MMPs) family such as MMP1, MMP2, MMP7 was down-regulated in transcriptional level. In conclusion, the combination of glycitin and 38 has induced the fibroblast proliferation even when it was damaged by UV exposure and H2O2, whereas augmented collagen production and inhibited the MMPs caused the wrinkle formation and decreased the melanocyte proliferation, suggested an potential UV-protective therapy.

Keywords: UV radiation, wrinkle, ageing, glycitin, dermal fibroblast

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Authors: Amina Abubakar

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English like any other language is rich by means of arbitrary, conventional, symbols which lend it to lot of inconsistencies in spelling, phonology, syntax, and morphology. The research examines the irregularities prevalent in the structure and meaning of some ‘er’ lexical items in English and its implication to vocabulary acquisition. It centers its investigation on the derivational suffix ‘er’, which changes the grammatical category of word. English language poses many challenges to Second Language Learners because of its irregularities, exceptions, and rules. One of the meaning of –er derivational suffix is someone or somebody who does something. This rule often confuses the learners when they meet with the exceptions in normal discourse. The need to investigate instances of such inconsistencies in the formation of –er words and the meanings given to such words by the students motivated this study. For this purpose, some senior secondary two (SS2) students in six randomly selected schools in the metropolis were provided a large number of alphabetically selected ‘er’ suffix ending words, The researcher opts for a test technique, which requires them to provide the meaning of the selected words with- er. The marking of the test was scored on the scale of 1-0, where correct formation of –er word and meaning is scored one while wrong formation and meaning is scored zero. The number of wrong and correct formations of –er words meaning were calculated using percentage. The result of this research shows that a large number of students made wrong generalization of the meaning of the selected -er ending words. This shows how enormous the inconsistencies are in English language and how are affect the learning of English. Findings from the study revealed that though students mastered the basic morphological rules but the errors are generally committed on those vocabulary items that are not frequently in use. The study arrives at this conclusion from the survey of their textbook and their spoken activities. Therefore, the researcher recommends that there should be effective reappraisal of language teaching through implementation of the designed curriculum to reflect on modern strategies of teaching language, identification, and incorporation of the exceptions in rigorous communicative activities in language teaching, language course books and tutorials, training and retraining of teachers on the strategies that conform to the new pedagogy.

Keywords: ESL(English as a second language), derivational morpheme, inflectional morpheme, suffixes

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Authors: Pavel Kotin, Sergey Dotofeev, Daniil Kozlov, Alexey Garshev

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We propose the investigation of CdSe quantum dots which were synthesized in the presence of silver halides. To understand a process of nanoparticle formation in more detail, we varied the silver halide amount in the synthesis and proposed a sampling during colloidal growth. The attempts were focused on the investigation of shape, structure and optical properties of nanoparticles. We used the colloidal method of synthesis. Cadmium oleate, tri-n-octylphosphine selenide (TOPSe) and AgHal in TOP were precursors of cadmium, selenium and silver halides correspondingly. The molar Ag/Cd ratio in synthesis was varied from 1/16 to 1/1. The sampling was basically realized in 20 sec, 5 min, and 30 min after the beginning of quantum dots nucleation. To investigate nanoparticles we used transmission electron microscopy (including high resolution one), X-ray diffraction, and optical spectroscopy. It was established that silver halides lead to obtaining tetrapods with different leg length and large ellipsoidal nanoparticles possessing an intensive near IR photoluminescence. The change of the amount of silver halide in synthesis and the selection of an optimal growth time allows controlling the shape and the share of tetrapods or ellipsoidal nanoparticles in the product. Our main attempts were focused on a detailed investigation of the quantum dots structure and shape evolution and, finally, on mechanisms of such nanoparticle formation.

Keywords: colloidal quantum dots, shape evolution, silver doping, tetrapods

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Authors: Hassan M. Baioumy, Yuniarti Ulfa

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Paleozoic, Mesozoic and Cenozoic black shales that can be a potential source of energy and precious metals are widely distributed in Malaysia Peninsula, Sarawak and Sabah. Two Paleozoic black shales outcrops were reported in the Langkawi Island belonging to the Cambrian fluvial Machinchang Formation and the Silurian glaciomarine Singa Formation. More the seventeen occurrences of Paleozoic black shales outcrops have been found in the Peninsular Malaysia that range in age from Devonian, Carboniferous, and Permian in the Terengganu, Perlis, Pahang, and Perak States. Mesozoic black shales outcrops occur in several places in both the Peninsular Malaysia and Sarawak. In the Peninsular Malaysia, Triassic black shales occur in the Nami area, Northern Kedah and in the Pahang area. In Sarawak, Triassic black shales have been reported in the Bau area. Cenozoic black shales outcrops were reported in both Sarawak at Miri area and Sabah at the Ranau and Tenom areas. Preliminary mineralogical and geochemical investigations on some of these black shales outcrops showed distinct compositional variations among these black shales outcrops probably due to variations in their source area composition and/or depositional and diagenetic settings of these shales. Some of these shalese also subjected to post-depositional hydrothermal mineralization that enriched these shales with Au-bearing minerals such as pyrite, calchopyrite, and arsenopyrite. Many of the studied black shales outcrops look rich in organic matter, which increase the possibility of using these black shales as an unconventional energy resource.

Keywords: black shales, energy, mineralization, Malaysia

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Authors: Ben Yahia Nouha, Harris Chris, Boussen Slim, Chaabani Fredj

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The present work has set itself the objective of studying non-detritic siliceous rocks in the extreme northwestern of Tunisia. It aims to discuss the origin and depositional environment of siliceous rocks based on petrographic, mineralogical, and geochemical results. The different sections were made in the area of Babouch and the area of Cap-Serrat. The collected samples were subjected to petrographic, mineralogical, and geochemical characterization using different analytical methods: scanning electron microscopy (SEM), X-ray diffraction (XRD), geochemical analysis (ICP- AES), isotopic geochemistry (δ¹⁸O) to assess their suitability for industrial use. These babouchite shows that the mineralogy consists of quartz as the dominant mineral with the total lack of amorphous silica, while clay represents the minor phase. The petrographic examination revealed allowed to deduce that it is a rock of chemical origin deriving from tests of siliceous organisms (the radiolarians). Chemical analyzes show that SiO₂, Al₂O₃, and Fe₂O₃ represent the most abundant oxides. The other oxides are present in negligible quantity. Geochemical data support a biogenic and non-hydrothermal origin of babouchite silica. Oxygen isotopic has shown that babouchites are formed in an environment with a high temperature, ranging from 56°C to 73°C.

Keywords: siliceous rocks, babouchite formation, XRD, chemical analysis, isotopic geochemistry, Northwestern of Tunisia

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Authors: Adela Diepoltova, Klara Konecna, Ondrej Jandourek, Petr Nachtigal

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A loss of control over antibiotic-resistant pathogens has become a global issue due to severe and often untreatable infections. This state is reflected in complicated treatment, health costs, and higher mortality. All these factors emphasize the urgent need for the discovery and development of new anti-infectives. One of the most common pathogens mentioned in the phenomenon of antibiotic resistance are bacteria of the genus Staphylococcus. These bacterial agents have developed several mechanisms against the effect of antibiotics. One of them is biofilm formation. In staphylococci, biofilms are associated with infections such as endocarditis, osteomyelitis, catheter-related bloodstream infections, etc. To author's best knowledge, no validated and standardized methodology evaluating candidate compound activity against staphylococcal biofilms exists. However, a variety of protocols for in vitro drug activity testing has been suggested, yet there are often fundamental differences. Based on our experience, a key methodological step that leads to credible results is to form a robust biofilm with appropriate attributes such as firm adherence to the substrate, a complex arrangement in layers, and the presence of extracellular polysaccharide matrix. At first, for the purpose of drug antibiofilm activity evaluation, the focus was put on various conditions (supplementation of cultivation media by human plasma/fetal bovine serum, shaking mode, the density of initial inoculum) that should lead to reproducible and robust in vitro staphylococcal biofilm formation in microtiter plate model. Three model staphylococcal reference strains were included in the study: Staphylococcus aureus (ATCC 29213), methicillin-resistant Staphylococcus aureus (ATCC 43300), and Staphylococcus epidermidis (ATCC 35983). The total biofilm biomass was quantified using the Christensen method with crystal violet, and results obtained from at least three independent experiments were statistically processed. Attention was also paid to the viability of the biofilm-forming staphylococcal cells and the presence of extracellular polysaccharide matrix. The conditions that led to robust biofilm biomass formation with attributes for biofilms mentioned above were then applied by introducing an alternative method analogous to the commercially available test system, the Calgary Biofilm Device. In this test system, biofilms are formed on pegs that are incorporated into the lid of the microtiter plate. This system provides several advantages (in situ detection and quantification of biofilm microbial cells that have retained their viability after drug exposure). Based on our preliminary studies, it was found that the attention to the peg surface and substrate on which the bacterial biofilms are formed should also be paid to. Therefore, further steps leading to the optimization were introduced. The surface of pegs was coated by human plasma, fetal bovine serum, and L-polylysine. Subsequently, the willingness of bacteria to adhere and form biofilm was monitored. In conclusion, suitable conditions were revealed, leading to the formation of reproducible, robust staphylococcal biofilms in vitro for the microtiter model and the system analogous to the Calgary biofilm device, as well. The robustness and typical slime texture could be detected visually. Likewise, an analysis by confocal laser scanning microscopy revealed a complex three-dimensional arrangement of biofilm forming organisms surrounded by an extracellular polysaccharide matrix.

Keywords: anti-biofilm drug activity screening, in vitro biofilm formation, microtiter plate model, the Calgary biofilm device, staphylococcal infections, substrate modification, surface coating

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