Search results for: purified bentonite

Authors: Wael M. Rabeh, Cesar Carrasco-Lopez, Juliana C. Ferreira, Pance Naumov

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Bioluminescence, the emission of light from a biological process, is found in various living organisms including bacteria, fireflies, beetles, fungus and different marine organisms. Luciferase is an enzyme that catalyzes a two steps oxidation of luciferin in the presence of Mg2+ and ATP to produce oxyluciferin and releases energy in the form of light. The luciferase assay is used in biological research and clinical applications for in vivo imaging, cell proliferation, and protein folding and secretion analysis. The luciferase enzyme consists of two domains, a large N-terminal domain (1-436 residues) that is connected to a small C-terminal domain (440-544) by a flexible loop that functions as a hinge for opening and closing the active site. The two domains are separated by a large cleft housing the active site that closes after binding the substrates, luciferin and ATP. Even though all insect luciferases catalyze the same chemical reaction and share 50% to 90% sequence homology and high structural similarity, they emit light of different colors from green at 560nm to red at 640 nm. Currently, the majority of the structural and biochemical studies have been conducted on green-emitting firefly luciferases. To address the color emission mechanism, we expressed and purified two luciferase enzymes with blue-shifted green and red emission from indigenous Brazilian species Amydetes fanestratus and Phrixothrix, respectively. The two enzymes naturally emit light of different colors and they are an excellent system to study the color-emission mechanism of luciferases, as the current proposed mechanisms are based on mutagenesis studies. Using a vapor-diffusion method and a high-throughput approach, we crystallized and solved the crystal structure of both enzymes, at 1.7 Å and 3.1 Å resolution respectively, using X-ray crystallography. The free enzyme adopted two open conformations in the crystallographic unit cell that are different from the previously characterized firefly luciferase. The blue-shifted green luciferase crystalized as a monomer similar to other luciferases reported in literature, while the red luciferases crystalized as an octamer and was also purified as an octomer in solution. The octomer conformation is the first of its kind for any insect’s luciferase, which might be relate to the red color emission. Structurally designed mutations confirmed the importance of the transition between the open and close conformations in the fine-tuning of the color and the characterization of other interesting mutants is underway.

Keywords: bioluminescence, enzymology, structural biology, x-ray crystallography

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Authors: Nuha Mansour Alhazmi

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Replacement of petro-based plastics by a biodegradable plastic are vastly growing process. Poly-β-hydroxybutyrate (PHB) is a biodegradable biopolymer, synthesized by some bacterial genera. The objective of the current study is to explore the ability of some fungi to biodegrade PHB. The degradation of (PHB) was detected in Petri dish by the formation of a clear zone around the fungal colonies due to the production of depolymerase enzyme which has an interesting role in the PHB degradation process. Among 10 tested fungi, the most active PHB biodegraded fungi were identified as Trichoderma asperellum using morphological and molecular characters. The highest PHB degradation was at 25°C, pH 7.5 after 7 days of incubation for the tested fungi. Finally, the depolymerase enzyme was isolated, purified using column chromatography and characterized. In conclusion, PHB can be biodegraded in solid and liquid medium using depolymerase enzyme from T. asperellum.

Keywords: degradation, depolymerase enzyme, PHB, Trichoderma asperellum

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Authors: C. Nobre, C. C. Castro, A.-L. Hantson, J. A. Teixeira, L. R. Rodrigues, G. De Weireld

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Fructo-oligosaccharides (FOS) are produced from sucrose by Aureobasidium pullulans in yields between 40-60% (w/w). To increase the amount of FOS it is necessary to remove the small, non-prebiotic sugars, present. Two methods for producing high-purity FOS have been developed: the use of microorganisms able to consume small saccharides; and the use of continuous chromatography to separate sugars: simulated moving bed (SMB). It is herein proposed the combination of both methods. The aim of this study is to optimize the composition of the fermentative broth (in terms of salts and sugars) that will be further purified by SMB. A yield of 0.63 gFOS.g Sucrose-1 was obtained with A. pullulans using low amounts of salts in the initial fermentative broth. By removing the small sugars, Saccharomyces cerevisiae and Zymomonas mobilis increased the percentage of FOS from around 56.0% to 83% (w/w) in average, losing only 10% (w/w) of FOS during the recovery process.

Keywords: fructo-oligosaccharides, microbial treatment, Saccharomyces cerevisiae, Zymomonas mobilis

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Authors: A. M. Isakhanyan, F. N. Tkhruni, N. N. Yakimovich, Z. I. Kuvaeva, T. V. Khachatryan

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Introduction: At present, the simbiotics based on different genera and species of lactic acid bacteria (LAB) and yeasts are used. One of the basic properties of probiotics is presence of antimicrobial activity and therefore selection of LAB and yeast strains for their co-cultivation with the aim of increasing of the activity is topical. Since probiotic yeast and bacteria have different mechanisms of action, natural synergies between species, higher viability and increasing of antimicrobial activity might be expected from mixing both types of probiotics. Endemic strains of LAB Enterococcus faecium БТK-64, Lactobaccilus plantarum БТK-66, Pediococcus pentosus БТK-28, Lactobacillus rhamnosus БТK-109 and Kluyveromyces lactis БТX-412, Saccharomycopsis sp. БТX- 151 strains of yeast, with probiotic properties and hight antimicrobial activity, were selected. Strains are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB and yeast strains were isolated from different dairy products from rural households of Armenia. The genotyping by 16S rRNA sequencing for LAB and 26S RNA sequencing for yeast were used. Combined cultivation of LAB and yeast strains was carried out in the nutrient media on the basis of milk whey, in anaerobic conditions (without shaker, in a thermostat at 37oC, 48 hours). The complex preparations were obtained by purification of cell free culture broth (CFC) broth by the combination of ion-exchange chromatography and gel filtration methods. The spot-on-lawn method was applied for determination of antimicrobial activity and expressed in arbitrary units (AU/ml). Results. The obtained data showed that at the combined growth of bacteria and yeasts, the cultivation conditions (medium composition, time of growth, genera of LAB and yeasts) affected the display of antimicrobial activity. Purification of CFC broth allowed obtaining partially purified antimicrobial complex preparation which contains metabiotics from both bacteria and yeast. The complex preparation inhibited the growth of pathogenic and conditionally pathogenic bacteria, isolated from various internal organs from diseased animals and poultry with greater efficiency than the preparations derived individually alone from yeast and LAB strains. Discussion. Thus, our data shown perspectives of creation of a new class of antimicrobial preparations on the basis of combined cultivation of endemic strains of LAB and yeast. Obtained results suggest the prospect of use of the partially purified complex preparations instead antibiotics in the agriculture and for food safety. Acknowledgments: This work was supported by the RA MES State Committee of Science and Belarus National Foundation for Basic Research in the frames of the joint Armenian - Belarusian joint research project 13РБ-064.

Keywords: co-cultivation, antimicrobial activity, biosafety, metabiotics, lactic acid bacteria, yeast

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Authors: E. Erdem, M. Şahin, M. Saçak

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Smectite is a layered silicate and modified with alkyl ammonium salts to make both the hydrophilic silicate surfaces organophilic, and to expand the clay layers. Thus, a nanocomposite structure can be formed enabling to enter various types of polymers between the layers. In this study, Na-smectite crystals were prepared by purification of bentonite. Benzyltributylammonium bromide (BTBAB) was used as a swelling agent. The mixing time and additive concentration were changed during the swelling process. It was determined that the 4 h of mixing time and 0.2 g of BTBAB were sufficient and the usage of higher amounts of salt did not increase the interlayer space between the clay layers. Then, the conductive poly(o-toluidine) (POT)/smectite nanocomposite was prepared in the presence of swollen Na-smectite using ammonium persulfate (APS) as oxidant in aqueous acidic medium. The POT content and conductivity of the prepared nanocomposite were systematically investigated as a function of polymerization conditions such as the treatment time of swollen smectite in monomer solution and o-toluidine/APS mol ratio. The POT content and conductivity of nanocomposite increased with increasing monomer/oxidant mol ratio up to 1 and did not change at higher ratios. The maximum polymer yield and the highest conductivity value of the composite were 26.0% and 4.0×10-5 S/cm, respectively. The structural and morphological analyses of the POT/smectite nanocomposite were carried out by XRD, FTIR and SEM techniques, respectively.

Keywords: clay, composite, conducting polymer, poly(o-anisidine)

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Authors: M. Hoseinnezhad, K. Gharanjig

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Two organic dyes comprising carbazole as the electron donors and cyanoacetic acid moieties as the electron acceptors were synthesized. The organic dye was prepared by standard reaction from carbazole as the starting material. To this end, carbazole was reacted with bromobenzene and further oxidation and reacted with cyanoacetic acid. The obtained organic dye was purified and characterized using differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance (¹HNMR), carbon nuclear magnetic resonance (¹³CNMR) and elemental analysis. The influence of heteroatom on carbazole donors and cyno substitution on the acid acceptor is evidenced by spectral and electrochemical photovoltaic experiments. Finally, light fastness properties for organic dye were investigated.

Keywords: dye-sensitized solar cells, indoline dye, nanostructure, oxidation potential, solar energy

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Authors: B. T. Naveen Kumar, Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, A. H. Shanthanagouda, Orawan Boodde, K. M. Shankar, Prakash Patil, Shubhkaramjeet Kaur

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Early mortality syndrome (EMS)/Acute Hepatopancreatic Necrosis Disease (AHPND) has emerged as a major obstacle for the shrimp farming around the world. It is caused by a strain of Vibrio parahaemolyticus. The possible preventive and control measure is, early and rapid detection of the pathogen in the broodstock, post-larvae and monitoring the shrimp during the culture period. Polymerase chain reaction (PCR) based early detection methods are good, but they are costly, time taking and requires a sophisticated laboratory. The present study was conducted to develop a simple, sensitive and rapid diagnostic farmer level kit for the reliable detection of AHPND in shrimp. A panel of monoclonal antibodies (MAbs) were raised against the recombinant Pir B protein (rPirB). First, an immunodot was developed by using MAbs G3B8 and Mab G3H2 which showed specific reactivity to purified r-PirB protein with no cross-reactivity to other shrimp bacterial pathogens (AHPND free Vibrio parahaemolyticus (Indian strains), V. anguillarum, WSSV, Aeromonas hydrophila, and Aphanomyces invadans). Immunodot developed using Mab G3B8 is more sensitive than that with the Mab G3H2. However, immunodot takes almost 2.5 hours to complete with several hands-on steps. Therefore, the flow-through assay (FTA) was developed by using a plastic cassette containing the nitrocellulose membrane with absorbing pads below. The sample was dotted in the test zone on the nitrocellulose membrane followed by continuos addition of five solutions in the order of i) blocking buffer (BSA) ii) primary antibody (MAb) iii) washing Solution iv) secondary antibody and v) chromogen substrate (TMB) clear purple dots against a white background were considered as positive reactions. The FTA developed using MAbG3B8 is more sensitive than that with MAb G3H2. In FTA the two MAbs showed specific reactivity to purified r-PirB protein and not to other shrimp bacterial pathogens. The FTA is simple to farmer/field level, sensitive and rapid requiring only 8-10 min for completion. Tests can be developed to kits, which will be ideal for use in biosecurity, for the first line of screening (at the port or pond site) and during monitoring and surveillance programmes overall for the good management practices to reduce the risk of the disease.

Keywords: acute hepatopancreatic necrosis disease, AHPND, flow-through assay, FTA, farmer level, immunodot, pond site, shrimp

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Authors: Ajuna B. Henry

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In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

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Authors: Young-Kwan Choi, Gang-Wook Shin, Sung-Taek Hong

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Water supply system consumes large amount of power load during water treatment and transportation of purified water. Many energy conserving high efficiency materials such as DC motor and LED light have recently been introduced to water supply system for energy conservation. This paper performed empirical analysis on BLDC, AC motors, and comparatively analyzed the change in power according to DC power supply ratio in order to conserve energy of a next-generation water treatment system called vertical water treatment system. In addition, a DC distribution system linked with photovoltaic generation was simulated to analyze the energy conserving effect of DC load.

Keywords: vertical water treatment system, DC power supply, energy efficiency, BLDC

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Authors: Devesh Motwani, Amey Kashyap

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Interest in cost effective drilling has increased substantially in the past years. Economics associated with drilling fluids is needed to be considered seriously for lesser cost per foot in planning and drilling of a wellbore and the various environmental concerns imposed by international communities related with the constituents of the drilling fluid. Viscofier such as Guar Gum is a high molecular weight polysaccharide from Guar plants, is used to increase viscosity in water-based and brine-based drilling fluids thus enabling more efficient cleaning of the bore. Other applications of this Viscofier are to reduce fluid loss by giving a better colloidal solution, decrease fluid friction and so minimising power requirements and used in hydraulic fracturing to increase the recovery of oil and gas. Guar gum is also used as a surfactant, synthetic polymer and defoamer. This paper presents experimental results to verifying the properties of guar gum as a viscofier and filtrate retainer as well as observing the impact of different quantities of guar gum and Carboxymethyl cellulose (CMC) in a standard sample of water based bentonite mud solution. This is in attempt to make a drilling fluid which contains half of the quantity of drilling mud used and yet is equally viscous to the standardised mud sample. Thus we can see that mud economics will be greatly affected by this approach. However guar gum is thermally stable till 60-65°C thus limited to be used in drilling shallow wells and for a wider thermal range, suitable chrome free additives are required.

Keywords: economics, guargum, viscofier, CMC, thermal stability

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Authors: M. Hosseinnejad, K. Gharanjig

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In the present study, metal free organic dyes were prepared and used as photo-sensitizers in dye-sensitized solar cells. Double rhodanine was utilized as the fundamental electron acceptor group to which electron donor aldehyde with varying substituents was attached to produce new organic dye. This dye was first purified and then characterized by analytical techniques. Spectrophotometric evaluations of the prepared dye in solution and on a nano anatase TiO₂ substrate were carried out in order to assess possible changes in the status of the dyes in different environments. The results show that the dye form j-type aggregates on the nano TiO₂. Additionally, oxidation potential measurements were also carried out. Finally, dye sensitized solar cell based on synthesized dye was fabricated in order to determine the photovoltaic behavior and conversion efficiency of individual dye.

Keywords: conversion efficiency, dye-sensitized solar cell, photovoltaic behavior, sensitizer

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Authors: Karima Ould Yerou, B. Meddah, A. Tir Touil

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The intestinal flora also called the intestinal microbiota, consists of different bacteria and other microorganisms which occur naturally in the gastrointestinal tract organs components. These intestinal bacteria are present in their millions and help the functioning of the body in particular allowing aid to degradation of certain molecules into absorbable substrates. They also protect against invasion of the gut by other pathogenic bacteria, that is to say which may be responsible for disease. Factors like stress, antibiotics and diet can affect the balance of intestinal flora and in case of imbalance, digestive disorders type bloating, diarrhea or vomiting may occur. Rattus norvegicus of bad weight of 100 kg, an Algerian firm West are scarified and isolation of their ileum and colon respectively two Bactrian strains Escherichia coli and Lactobacillus are then purified and identified.

Keywords: intestinal flora, Rattus norvegicus, Escherichia coli, lactobacillus, West Algerian farm

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Authors: Sabiha Chouchane, Toufik Chouchane, Azzedine Hani

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The wastewater was treated with a natural asorbent “Diatomite” to eliminate chromium three. Diatomite is an element that comes from Sig (west of Algeria). The physicochemical characterization revealed that the diatomite is mainly made up of silica, lime and a lower degree of alumina. The process considered in static regime, at 20°C, an ion stirring speed of 150 rpm, a pH = 4 and a grain diameter of between 100 and 150µm, shows that one gram of diatomite purified can fix according to the Langmuir model up to 39.64 mg/g of chromium with pseudo 1st order kinetics. The pseudo-equilibrium time highlighted is 25 minutes. The affinity between the adsorbent and the adsorbate follows the value of the RL ratio indicates us that the solid used has a good adsorption capacity. The external transport of the metal ions from the solution to the adsorbent seems to be a step controlling the speed of the overall process. On the other hand, internal transport in the pores is not the only limiting mechanism of sorption kinetics. Thermodynamic parameters show that chromium sorption is spontaneous and exothermic with negative entropy.

Keywords: adsorption, diatomite, crIII, wastewater

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Authors: N. Zeinalzadeh, Mahdi Sadeghi

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Enterotoxigenic Escherichia coli (ETEC) is the most common cause of non-inflammatory diarrhea in the developing countries, resulting in approximately 20% of all diarrheal episodes in children in these areas. ST is one of the most important virulence factors and CFA/I is one of the frequent colonization factors that help to process of ETEC infection. ST and CfaB (CFA/I subunit) are among vaccine candidates against ETEC. So, ST because of its small size is not a good immunogenic in the natural form. However to increase its immunogenic potential, here we explored candidate positions for ST insertion in CfaB sequence. After bioinformatics analysis, one of the candidate positions was selected and the chimeric gene (cfaB*st) sequence was synthesized and expressed in E. coli BL21 (DE3). The chimeric recombinant protein was purified with Ni-NTA columns and characterized with western blot analysis. The residue 74-75 of CfaB sequence could be a good candidate position for ST and other epitopes insertion.

Keywords: bioinformatics, CFA/I, enterotoxigenic E. coli, ST toxoid

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Authors: Khodzhaberdi Allaberdiev

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In order to understand the interactions on the interface polyamide fibers and epoxy matrix in fiber- reinforced composites were investigated industrial aramid fibers: armos, svm, terlon using individual epoxy matrix components, epoxies: diglycidyl ether of bisphenol A (DGEBA), three- and diglycidyl derivatives of m, p-amino-, m, p-oxy-, o, m,p-carboxybenzoic acids, the models: curing agent, aniline and the compound, that depict of the structure the primary addition reaction the amine to the epoxy resin, N-di (oxyethylphenoxy) aniline. The chemical structure of the surface of untreated and treated polyamide fibers analyzed using Fourier transform infrared spectroscopy (FTIR). The impregnation of fibers with epoxy matrix components and N-di (oxyethylphenoxy) aniline has been carried out by heating 150˚C (6h). The optimum fiber loading is at 65%.The result a thermal treatment is the covalent bonds formation , derived from a combined of homopolymerization and crosslinking mechanisms in the interfacial region between the epoxy resin and the surface of fibers. The reactivity of epoxy resins on interface in microcomposites (MC) also depends from processing aids treated on surface of fiber and the absorbance moisture. The influences these factors as evidenced by the conversion of epoxy groups values in impregnated with DGEBA of the terlons: industrial, dried (in vacuum) and purified samples: 5.20 %, 4.65% and 14.10%, respectively. The same tendency for svm and armos fibers is observed. The changes in surface composition of these MC were monitored by X-ray photoelectron spectroscopy (XPS). In the case of the purified fibers, functional groups of fibers act as well as a catalyst and curing agent of epoxy resin. It is found that the value of the epoxy groups conversion for reinforced formulations depends on aromatic polyamides nature and decreases in the order: armos >svm> terlon. This difference is due of the structural characteristics of fibers. The interfacial interactions also examined between polyglycidyl esters substituted benzoic acids and polyamide fibers in the MC. It is found that on interfacial interactions these systems influences as well as the structure and the isomerism of epoxides. The IR-spectrum impregnated fibers with aniline showed that the polyamide fibers appreciably with aniline do not react. FTIR results of treated fibers with N-di (oxyethylphenoxy) aniline fibers revealed dramatically changes IR-characteristic of the OH groups of the amino alcohol. These observations indicated hydrogen bondings and covalent interactions between amino alcohol and functional groups of fibers. This result also confirms appearance of the exo peak on Differential Scanning Calorimetry (DSC) curve of the MC. Finally, the theoretical evaluation non-covalent interactions between individual epoxy matrix components and fibers has been performed using the benzanilide and its derivative contaning the benzimidazole moiety as a models of terlon and svm,armos, respectively. Quantum-topological analysis also demonstrated the existence hydrogen bond between amide group of models and epoxy matrix components.All the results indicated that on the interface polyamide fibers and epoxy matrix exist not only covalent, but and non-covalent the interactions during the preparation of MC.

Keywords: epoxies, interface, modeling, polyamide fibers

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Authors: Yasser M. Saad, Heba El-Sebaie Abd El-Sadek

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Some Metapenaeus monoceros cox1 gene fragments were isolated, purified, sequenced, and comparatively analyzed with some other Crustacean Cox1 gene sequences (obtained from National Center for Biotechnology Information). This work was designed for testing the efficiency of this system in reconstruction of phylogenetic relations among some Crustacean species belonging to four genera (Metapenaeus, Artemia, Daphnia and Calanus). The single nucleotide polymorphism and haplotype diversity were calculated for all estimated mt-DNA fragments. The genetic distance values were 0.292, 0.015, 0.151, and 0.09 within Metapenaeus species, Calanus species, Artemia species, and Daphnia species, respectively. The reconstructed phylogenetic tree is clustered into some unique clades. Cytochrome oxidase subunit 1 gene (cox1) was a powerful system in reconstruction of phylogenetic relations among evaluated crustacean species.

Keywords: crustaceans, genetics, Cox1, phylogeny

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Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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Authors: Wang Yanping Karen, Mohd Rafeah Siti, Park MI Kyoung

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Formalin-fixed paraffin embedded (FFPE) tissue is important in the area of oncological diagnostics. This method of preserving tissues enabling them to be stored easily at ambient temperature for a long time. This decreases the risk of losing the DNA quantity and quality after extraction, reducing sample wastage, and making FFPE more cost effective. However, extracting DNA from FFPE tissue is a challenge as DNA purified is often highly cross-linked, fragmented, and degraded. In addition, this causes problems for many downstream processes. In this study, there will be a comparison of DNA extraction efficiency between One BioMed’s Xceler8 automated platform with commercial available extraction kits (Qiagen and Roche). The FFPE tissue slices were subjected to deparaffinization process, pretreatment and then DNA extraction using the three mentioned platforms. The DNA quantity were determined with real-time PCR (BioRad CFX ) and gel electrophoresis. The amount of DNA extracted with the One BioMed’s X8 platform was found to be comparable with the other two manual extraction kits.

Keywords: DNA extraction, FFPE tissue, qiagen, roche, one biomed X8

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Authors: Ait Slimane-Ait Kaki Sabrina, Foudi Lamia

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The present work consists of the study of the bio-ecology and the therapeutic effects of the belladone (Atropa belladonna L.). It is a medicinal plant of the Solanacées family, herbaceous, robust 0.5 up to 1.50 m high. The phytochemical analysis of leaves revealed alkaloids, tannins, catechin, coumarins, mucilages, saponins, starch, and reducing compounds. The experimental study concerns the extraction and characterization of belladonna alkaloids. Analysis of the purified extract by staining tests confirmed the presence of tropane alkaloids. The dosage chromatography revealed the presence of components that have been identified atropine, scopolamine and hyoscyamine. Evaluation of antimicrobial and antifungal alkaloids from the methanol extract and aqueous extract of belladonna on pathogenic germs showed a positive bactericidal against strains of Escherichia coli and Staphylococcus aureus. Our preliminary results allow us an overall assessment of the medicinal value of Atropa belladonna.

Keywords: belladone, alkaloid, antibacterial activity, antifungal activity

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Authors: Dalila Oulhaci, Mohammed Zahaf

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The watering technique is essential to maintain a moist perimeter around the roots of the crop. This is the case with topical watering, where the soil around the root system can be kept permanently moist between the two extremes of water content. Moreover, one of the oldest methods used since Roman times throughout North Africa and the Near East was based on the repeated pouring of water into porous earthen vessels buried in the ground. In this context, these two techniques have been combined by replacing the earthen vase with plastic bottles filled with sand which release water through their perforated walls into the surrounding soil. The objective of this work is to first determine the purifying power of the activated sludge treatment plant of Toggourt and then that of the bottled Sawdust filter. For the station, the BOD purification rate was (96.5%), the COD purification rate was (87%) and suspended solids (90%). For the bottle, the BOD removal rate was (35%), and COD removal rate was (12.58%). This work falls within the framework of water saving, sustainable development and environmental protection, and also within the framework of agriculture.

Keywords: wasterwater, sawdust, purification, irrigation, touggourt (Algeria)

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Authors: Esma Kocaoglu, Oktay Talaz, Huseyin Cavdar, Murat Senturk, Deniz Eki̇nci̇

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Glutathione reductase (GR) is a crucial antioxidant enzyme which is responsible for the maintenance of the antioxidant GSH (glutathione) molecule. Antimalarial effects of some chemical molecules are attributed to their inhibition of GR; thus inhibitors of this enzyme are expected to be promising candidates for the treatment of malaria. In this work, GR inhibitory properties of N-Methylpyrrole derivatives are reported. Firstly, GR was purified by means of affinity chromatography using 2’,5’-ADP-Sepharose 4B as ligand. Enzymatic activity was measured by Beutler’s method. Synthesis of the compounds was approved by thin layer chromatography and column chromatography. Different inhibitor concentrations were used and all compounds were tested in triplicate at each concentration used. It was found that all compounds have better inhibitory activity than the strong GR inhibitor N,N-bis(2-chloroethyl)-N-nitrosourea, especially three molecules, 8m, 8n, and 8q, are the best among them with low micromolar I₅₀ values. Findings of our study indicate that these Schiff base derivatives are strong GR inhibitors which can be used as leads for designation of novel antimalaria candidates.

Keywords: glutathione reductase, antimalaria, inhibitor, enzyme

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Authors: Imad H. Hameed, Mohammad A. Jebor, Ammera J. Omer

Abstract:

The aim of this research is to study the mitochonderial coding region by using the Sanger sequencing technique and establish the degree of variation characteristic of a fragment. FTA® Technology (FTA™ paper DNA extraction) utilized to extract DNA. Portion of coding region encompassing positions 11719 –12384 amplified in accordance with the Anderson reference sequence. PCR products purified by EZ-10 spin column then sequenced and Detected by using the ABI 3730xL DNA Analyzer. Five new polymorphic positions 11741, 11756, 11878, 11887 and 12133 are described may be suitable sources for identification purpose in future. The calculated value D= 0.95 and RMP=0.048 of the genetic diversity should be understood as high in the context of coding function of the analysed DNA fragment. The relatively high gene diversity and a relatively low random match probability were observed in Iraq population. The obtained data can be used to identify the variable nucleotide positions characterized by frequent occurrence which is most promising for various identifications.

Keywords: coding region, Iraq, mitochondrial DNA, polymorphic positions, sanger technique

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Authors: Iannello G., Coppa F., Pennisi S., Giuffrida G., Lo Faro R., Cartelli S., Ferruggia G., Brundo M. V.

Abstract:

Skin is the largest multifunctional human organ and possesses a complex, multilayered structure with the ability to regenerate and renew. The key role in skin regeneration is played by fibroblasts, which also occupy an important role in the wound healing process. Different methods, including dynamic light scattering, scanning electron microscopy, ELISA, and MTT assay, were employed to evaluate on fibroblasts the in vitro effects of plant-derived nanovesicles and cord blood stem cells‐derived exosomes. We compared the results with those of cells exposed to a technology called AMPLEX PLUS, containing a mixture of 20 different biologically active factors (GF20) and exosomes isolated and purified from bovine colostrum. AMPLEX PLUS was able to significantly enhance the cell proliferation status of cells at both 24 and 48 hours compared to untreated cells (control). The obtained results suggest how AMPLEX PLUS could be potentially effective in treating skin rejuvenation.

Keywords: AMPLEX PLUS, cell vitality, colostrum, nanovesicles

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Authors: Chun-Wei Liu, Feng-Tsai Weng

Abstract:

People faced pollution threaten in modern age although the standard of exhaust gas of vehicles has been established. The goal of this study is to investigate the effect of water atomizer in a vehicle emission system. Diluted 20% ammonia water was used in spraying system. Micro particles produced by exhausted gas from engine of vehicle which were cumulated through atomized spray in a self-development collector. In experiments, a self-designed atomization model plate and a gas tank controlled by the micro-processor using Pulse Width Modulation (PWM) logic was prepared for exhaust test. The gas from gasoline-engine of vehicle was purified with the model panel collector. A soft well named ANSYS was utilized for analyzing the distribution condition of rejected gas. Micro substance and percentage of CO, HC, CO2, NOx in exhausted gas were investigated at different engine speed, and atomizer vibration frequency. Exceptional results in the vehicle engine emissions measurement were obtained. The temperature of exhausted gas can be decreased 3oC. Micro substances PM10 can be decreased and the percentage of CO can be decreased more than 55% at 2500RPM by proposed system. Value of CO, HC, CO2 and NOX was all decreased when atomizers were used with water.

Keywords: atomizer, CO, HC, NOx, PM2.5

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Authors: Ankur Sachan

Abstract:

Carbon dioxide being the most anthropogenic greenhouse gas,it needs to be isolated from entering into atmosphere. Carbon capture and storage is process that captures CO2 emitted from various sources, separates it from other gases and stores it in a safe place preferably in underground geological formations for large period of time. It is then purified and monitored so that can be made to reuse. Monoethanolamine, zeolitic imidazolate framework, microalgae, membranes etc are utilized to capture CO2. Post-combustion, pre-combustion and oxyfuel combustion along with chemical looping combustion are technologies for scrubbing CO2. The properties of CO2 being easily miscible and readily dissolving in oil with impurities makes it capable for numerous applications such as in producing oil by enhanced oil recovery (EOR), Bio CCS Algal Synthesis etc. CO2-EOR operation is capable to produce million barrels of oil and extend the field's lifetime as in case of Weyburn Oil Field in Canada. The physical storage of CO2 is technically the most feasible direction provided that the associated safety and sustainability issues can be met and new materials for CCUS process at low cost are urgently found so that so that fossil based systems with carbon capture are cost competitive.

Keywords: carbon capture, CCUS, sustainability, oil

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Authors: M. Hosseinnejad, K. Gharanjig, S. Moradian

Abstract:

A novel metal free organic dyes based on indigo was prepared and used as sensitizers in dye-sensitized solar cells. The synthesized dye together with its corresponding intermediates were purified and characterized by analytical techniques. Such techniques confirmed the corresponding structures of dye and its intermediate and the yield of all the stages of dye preparation were calculated to be above 85%. Fluorometric analyses show fluorescence in the green region of the visible spectrum for dye. Oxidation potential measurements for dye ensured an energetically permissible and thermodynamically favourable charge transfer throughout the continuous cycle of photo-electric conversion. Finally, dye sensitized solar cells were fabricated in order to determine the photovoltaic behaviour and conversion efficiencies of dye. Such evaluations demonstrate rather medium conversion efficiencies of 2.33% for such simple structured synthesized dye. Such conversion efficiencies demonstrate the potentiality of future use of such dye structures in dye-sensitized solar cells with respect to low material costs, ease of molecular tailoring, high yields of reactions, high performance and ease of recyclability.

Keywords: conversion efficiency, Dye-sensitized solar cells, indigo, photonic material

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Authors: Ratna Siti Khodijah, Mirzam Abdurrachman

Abstract:

Gold is recovered from gold ores. Within the ores, there are not only gold but also several types of precious metals. Copper, silver, and platinum group elements (ruthenium, rhodium, palladium, rhenium, osmium, and iridium) are metals commonly found in the ores. These metals combine to form an ore because they have the same properties. It is due to their position in periodic-system-of-elements are near to gold. However, the presence of mercury in every gold ore has not been mentioned, even though it is located right next to gold in the periodic-system-of-elements and they are located in the same block, d-block. Thus, it is possible that mercury is contained in the ores. Moreover, the elements of the same group with mercury—zinc and cadmium—sometimes can be found in the ores. It is suspected that mercury can not be detected because the processing of gold ores usually using fire assay method. Before the ores melting, mercury would evaporate because it has the lowest boiling point of all precious metal in the ores. Therefore, it suggested doing research on the presence of mercury in gold ores by CVAAS method. The results of this study would obtain the amount of mercury in gold ores that should be purified. So it can be produced economically if possible.

Keywords: boiling point, d-block, fire assay, precious metal

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Authors: Nurhuda Manshoor, Mohd Fazirulrahman Fathil, Muhammad Hakim Jaafar, Mohd Amirul S. A. Jalil

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The leaves of Neobalanocarpus heimii were investigated for their oligostilbene contents. Prior to isolation process, the determinations of compounds were based on mass spectrometric fragmentation patterns. Three compounds, heimiol B, hopeaphenol, and vaticaphenol A were identified directly from the crude extract. Preparative high-performance liquid chromatography (HPLC) was used to isolate and purify the other compounds. The purified compounds were then analyzed using NMR spectroscopy to identify the compound structure and stereochemistry. The method employed for the research modified to comply with different HPLC techniques such as preparative and analytical techniques. The crude sample was injected into preparative HPLC to obtain several fractions which consist of oligostilbene mixture. The fractions were further isolated using analytical HPLC to obtain four pure compounds. The compounds then were characterized using nuclear magnetic resonance (NMR). The result shows that the leaves extract of Neobalanocarpus heimii contain three oligostilbenes, namely vaticanol A, balanocarpol, and vaticaphenol A, and a galactopyranose.

Keywords: balanocarpol, hemiol B, hopeaphenol, vaticanol A, vaticaphenol A

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Authors: Shumani Ramuhaheli, Christopher C. Enweremadu, Hilary L. Rutto

Abstract:

In this study, biodiesel from used cooking oil was produced as purified by washing with water (water wash) and amberlite (dry wash). The work presents the results of short term tests on performance characteristics of diesel engine using both biodiesel-fuel samples. In this investigation, the water wash biodiesel and dry wash biodiesel and diesel were compared for performance using a four-cylinder diesel engine. The torque, brake power, specific fuel consumption and brake thermal efficiency were analyzed. The tests showed that in all cases, dry wash biodiesel performed marginally poorer compared to water wash biodiesel. Except for brake thermal efficiency, diesel fuel had better engine performance characteristics compared to the biodiesel-fuel samples. According to these results, dry washing of biodiesel has a marginal effect on engine performance.

Keywords: biodiesel, engine performance, used cooking oil, water wash, dry wash

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Authors: Benedict O. Ayomanor, Cookey Iyen, Ifeoma S. Iyen

Abstract:

Rice hull (RH) biomass product gives feasible silica for exact temperature and period. The minimal fabrication price turns its best feasible produce to metallurgical grade silicon (MG-Si). In this work, to avoid ecological worries extending from CO₂ release to oil leakage on water and land, or nuclear left-over pollution, all finally add to the immense topics of ecological squalor; high purity silicon > 98.5% emerge set from rice hull ash (RHA) by solid-liquid removal. The RHA derived was purified by nitric and hydrochloric acid solutions. Leached RHA sieved, washed in distilled water, and desiccated at 1010ºC for 4h. Extra cleansing was achieved by carefully mixing the SiO₂ ash through Mg dust at a proportion of 0.9g SiO₂ to 0.9g Mg, galvanised at 1010ºC to formula magnesium silicide. The solid produced was categorised by X-ray fluorescence (XRF), X-ray diffractometer (XRD), and Fourier transformation infrared (FTIR) spectroscopy. Elemental analysis using XRF found the percentage of silicon in the material is approximately 98.6%, main impurities are Mg (0.95%), Ca (0.09%), Fe (0.3%), K (0.25%), and Al (0.40%).

Keywords: siliceous, leached, biomass, solid-liquid extraction

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