Search results for: fatty acid methyl esters

Authors: Sakshi Piplani, Ajit Kumar

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Valproic acid (VPA) is the first synthetic therapeutic agent used to treat epileptic disorders, which account for affecting nearly 1% world population. Teratogenicity caused by VPA has prompted the search for next generation drug with better efficacy and lower side effects. Recent studies have posed HDAC8 as direct target of VPA that causes the teratogenic effect in foetus. We have employed molecular dynamics (MD) and docking simulations to understand the binding mode of VPA and their analogues onto HDAC8. A total of twenty 3D-structures of human HDAC8 isoforms were selected using BLAST-P search against PDB. Multiple sequence alignment was carried out using ClustalW and PDB-3F07 having least missing and mutated regions was selected for study. The missing residues of loop region were constructed using MODELLER and energy was minimized. A set of 216 structural analogues (>90% identity) of VPA were obtained from Pubchem and ZINC database and their energy was optimized with Chemsketch software using 3-D CHARMM-type force field. Four major neurotransmitters (GABAt, SSADH, α-KGDH, GAD) involved in anticonvulsant activity were docked with VPA and its analogues. Out of 216 analogues, 75 were selected on the basis of lower binding energy and inhibition constant as compared to VPA, thus predicted to have anti-convulsant activity. Selected hHDAC8 structure was then subjected to MD Simulation using licenced version YASARA with AMBER99SB force field. The structure was solvated in rectangular box of TIP3P. The simulation was carried out with periodic boundary conditions and electrostatic interactions and treated with Particle mesh Ewald algorithm. pH of system was set to 7.4, temperature 323K and pressure 1atm respectively. Simulation snapshots were stored every 25ps. The MD simulation was carried out for 20ns and pdb file of HDAC8 structure was saved every 2ns. The structures were analysed using castP and UCSF Chimera and most stabilized structure (20ns) was used for docking study. Molecular docking of 75 selected VPA-analogues with PDB-3F07 was performed using AUTODOCK4.2.6. Lamarckian Genetic Algorithm was used to generate conformations of docked ligand and structure. The docking study revealed that VPA and its analogues have more affinity towards ‘hydrophobic active site channel’, due to its hydrophobic properties and allows VPA and their analogues to take part in van der Waal interactions with TYR24, HIS42, VAL41, TYR20, SER138, TRP137 while TRP137 and SER138 showed hydrogen bonding interaction with VPA-analogues. 14 analogues showed better binding affinity than VPA. ADMET SAR server was used to predict the ADMET properties of selected VPA analogues for predicting their druggability. On the basis of ADMET screening, 09 molecules were selected and are being used for in-vivo evaluation using Danio rerio model.

Keywords: HDAC8, docking, molecular dynamics simulation, valproic acid

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Authors: Nancy Awuor Oduor, Nils Moosdorf

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Harmful Algal Blooms (HABs) are a threat to coastal water quality, marine biodiversity, and human health. The attention on HABs and associated phycotoxins is still very low in tropical coastal developing countries despite the high dependence of local communities on coastal and marine resources for food and livelihoods and the growing evidence of the global increase in HABs frequency, toxicity, and geographical expansion. Lack of HABs monitoring thus creates a high risk of exposure due to uncertainty. This study assessed the spatial and temporal variability and effects of potential HAB-forming species in Kenya’s coastal waters. The preliminary results from 463 sampled collected over a series of 10 coastal surveys conducted over 267 Km of Kenya’s coastline between August 2021 and July 2022 revealed the presence of 87 potential algal blooming species belonging to 47 genera dominated by species capable of producing toxins, causing physical harm and high biomass at 41, 31 and 21 % respectively. The taxonomic composition was also dominated by dinoflagellates at 47%, followed by diatoms, cyanobacteria, and silicoflagellates at 39, 12, and 2%, respectively. About 92 % of the toxin-producing species were established in the creek waters. However, there were no significant variations established in species richness between the dry and wet seasons. Paralytic Shellfish Poisoning (PSP) toxin-producing dinoflagellates Alexandrium spp., Aphanizomenon spp., Gonyaulax spp., Gymnodinium spp., and Brachydinium capitatum, and Amnesic Shellfish Poisoning (ASP) Toxin producing diatoms Amphora spp., Nitzschia spp. and Pseudo-nitzschia spp. Frequented the area in low cell densities ranging between 5 and 1500 cells/L. However, no domoic acid (DA) and saxitoxins (SXTs) were detected during the July surveys. This does not mean that the toxins are absent in the area, and longer studies are recommended.

Keywords: harmful algal blooms, phycotoxins, saxitoxin, domoic acid, Kenya

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Authors: Berkas Khaoula

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Based on selection characteristics, high-density polyethylene (HDPE) extruded pipes are among the most economical and durable materials as well-designed solutions for water and gas transmission systems. The main reasons for such a choice are the high quality-performance ratio and the long-term service durability under aggressive conditions. Due to inevitable interactions with soils of different chemical compositions and transported fluids, aggressiveness becomes a key factor in studying resilient strength and life prediction limits. This phenomenon is known as environmental stress cracking resistance (ESCR). In this work, the effect of 3 acidic environments (5% acetic, 20% hydrochloric and 20% sulfuric) on HDPE-100 samples (~10x11x24 mm3). The results presented in the form (Δm/m0, %) as a function of √t indicate that the absorption, in the case of strong acids (HCl and H2SO4), evolves towards negative values involving material losses such as antioxidants and some additives. On the other hand, acetic acid and deionized water (DW) give a form of linear Fickean (LF) and B types, respectively. In general, the acids cause a slow but irreversible alteration of the chemical structure, composition and physical integrity of the polymer. The DW absorption is not significant (~0.02%) for an immersion duration of 69 days. Such results are well accepted in actual applications, while changes caused by acidic environments are serious and must be subjected to particular monitoring of the OIT factor (Oxidation Induction Time). After 55 days of aging, the H2SO4 and HCl media showed particular values with a loss of % mass in the interval [0.025-0.038] associated with irreversible chemical reactions as well as physical degradations. This state is usually explained by hydrolysis of the polymer, causing the loss of functions and causing chain scissions. These results are useful for designing and estimating the lifetime of the tube in service and in contact with adverse environments.

Keywords: HDPE, environmental stress cracking, absorption, acid media, chemical aging

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Authors: Valeriu Mihai But, Sorana Daniela Bolboacă, Adriana Elena Bulboacă

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The nutraceutical compound Curcumin (Curcuma longa L.) is known for its anti-inflammatory, anti-cancer, and antioxidant effects. This study aimed to evaluate the antioxidative and analgesic effects of Curcumin (CC) compared to Tramadol (T) in chemical-induced nociceptive pain in rats. Thirty-five rats were randomly divided into five groups of seven rats each and were treated as follows: C group (control group): treated with saline solution 0.9%, (1 ml, i.p. administration), ethanoic acid (EA) group: pretreated with saline solution 0.9% - 30 min before EA nociceptive pain induction, (1 ml, i.p. administration), T group: pretreated with Tramadol, 10 mg/kg body weight (bw), i.p. administration - 30 min before EA nociceptive pain induction, CC1-group: pretreated with 1 mg/100g bw Curcumin i.p. administration - 2 days before EA pain induction and CC2-group: pretreated with Curcumin 2 mg/100g bw i.p. administration - 2 days before EA nociceptive pain induction. The following oxidative stress parameters were assessed: malondialdehyde (MDA), nitric oxide (NOx), total oxidative status (TOS), total antioxidative capacity (TAC), and thiol (Th). The antalgic activity was measured by the ethanoic acid writhing test. Treatment with Curcumin, both 1 mg/100g bw, and 2 mg/100g bw, showed significant differences as compared with the control group (p<0.001) regarding malondialdehyde (MDA), nitric oxide (NOx), and total oxidative status (TOS) oxidative biomarkers. Pretreatment with 2 mg/100g bw of Curcumin presented a significant decrease in MDA values compared with Tramadol (p<0.001). The TAC significantly increased in pretreatment with Curcumin compared with group control. (p<0.001) The nociceptive response to EA was significantly reduced in Curcumin and Tramadol groups. Treatment with Curcumin at a higher concentration was more effective. In an experimental pain model, this study demonstrates an important antioxidant and antinociceptive activity of Curcumin comparable with Tramadol treatment.

Keywords: curcumin, nociception, oxidative stress, pain

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Authors: Ivna Mororó, Lise P. Labéjof, Stephanie Ribeiro, Kely Almeida

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Lipid droplets (LDs) are normally presented in greater or lesser number in the cytoplasm of almost all eukaryotic and some prokaryotic cells. They are independent organelles composed of a lipid ester core and a surface phospholipid monolayer. As a lipid storage form, they provide an available source of energy for the cell. Recently it was demonstrated that they play an important role in other many cellular processes. Among the many unresolved questions about them, it is not even known how LDs is formed, how lipids are recruited to LDs and how they interact with the other organelles. Excess fat in the organism is pathological and often associated with the development of some genetic, hormonal or behavioral diseases. The formation and accumulation of lipid droplets in the cytoplasm can be increased by exogenous physical or chemical agents. It is well known that ionizing radiation affects lipid metabolism resulting in increased lipogenesis in cells, but the details of this process are unknown. To better understand the mode of formation of LDs in liver cells, we investigate their ultrastructural morphology after irradiation. For that, Wistar rats were exposed to whole body gamma radiation from 60-cobalt at various single doses. Samples of the livers were processed for analysis under a conventional transmission electron microscope. We found that when compared to controls, morphological changes in liver cells were evident at the higher doses of radiation used. It was detected a great number of lipid droplets of different sizes and homogeneous content and some of them merged each other. In some cells, it was observed diffused LDs, not limited by a monolayer of phospholipids. This finding suggests that the phospholipid monolayer of the LDs was disrupted by ionizing radiation exposure that promotes lipid peroxydation of endo membranes. Thus the absence of the phospholipid monolayer may prevent the realization of some cellular activities as follow: - lipid exocytosis which requires the merging of LDs membrane with the plasma membrane; - the interaction of LDs with other membrane-bound organelles such as the endoplasmic reticulum (ER), the golgi and mitochondria and; - lipolysis of lipid esters contained in the LDs which requires the presence of enzymes located in membrane-bound organelles as ER. All these impediments can contribute to lipid accumulation in the cytoplasm and the development of diseases such as liver steatosis, cirrhosis and cancer.

Keywords: radiobiology, hepatocytes, lipid metabolism, transmission electron microscopy

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Authors: Mohd Adil, Rosina Khan, Danishuddin, Asad U. Khan

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The aim of this study was to evaluate the influence of the crude and active solvent fraction of Trachyspermum ammi on S. mutans cariogenicity, effect on expression of genes involved in biofilm formation and caries development in rats. GC–MS was carried out to identify the major components present in the crude and the active fraction of T. ammi. The crude extract and the solvent fraction exhibiting least MIC were selected for further experiments. Scanning electron microscopy was carried out to observe the effect of the extracts on S. mutans biofilm. Comparative gene expression analysis was carried out for nine selected genes. 2-Isopropyl-5-methyl-phenol was found as major compound in crude and the active fraction. Binding site of this compound within the proteins involved in biofilm formation was mapped with the help of docking studies. Real-time RT-PCR analyses revealed significant suppression of the genes involved in biofilm formation. All the test groups showed reduction in caries (smooth surface as well as sulcal surface caries) in rats. Moreover, it also provides new insight to understand the mechanism influencing biofilm formation in S. mutans. Furthermore, the data suggest the putative cariostatic properties of T. Ammi and hence can be used as an alternative medicine to prevent caries infection.

Keywords: bio-film, Streptococcus mutans, dental caries, bio-informatic

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Authors: Y. Kalachyova, O. Lyutakov, V. Svorcik

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One of the most significant obstacles for the application of surface enhanced Raman spectroscopy (SERS) is the poor reproducibility of SERS active substrates: SERS intensity can be varied from one substrate to another and moreover along the one substrate surface. High enhancement of the near-field intensity is the key factor for ultrasensitive SERS realization. SERS substrate can be prepared through introduction of highly ordered metal array, where light focusing is achieved through excitation of surface plasmon-polaritons (SPPs). In this work, we report the preparation of silver nanostructures with plasmon absorption peaks tuned by the metal arrangement. Excimer laser modification of poly(methyl methacrylate) followed by silver evaporation is proposed as an effective way for the creation of reproducible and effective surface plasmon-polaritons (SPP)-based SERS substrate. Theoretical and experimental studies were performed to optimize structure parameter for effective SPP excitation. It was found that the narrow range of grating periodicity and metal thickness exist, where SPPs can be most efficiently excited. In spite of the fact, that SERS response was almost always achieved, the enhancement factor was found to vary more with the effectivity of SPP excitation. When the real structure parameters were set to optimal for SPP excitation, a SERS enhancement factor was achieved up to four times. Theoretical and experimental investigation of SPP excitation on the two-dimensional periodical silver array was performed with the aim to make SERS response as high as possible.

Keywords: grating, nanostructures, plasmon-polaritons, SERS

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Authors: Henry O. Udeha, Kwaku G. Duodub, Afam I. O. Jideanic

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Finger millet (FM) [Eleusine coracana] is considered as a potential ‘‘super grain’’ by the United States National Academies as one of the most nutritious among all the major cereals. The regular consumption of FM-based diets has been associated with reduced risk of diabetes, cataract and gastrointestinal tract disorder. Hyperglycaemic, hypocholesterolaemic and anticataractogenic, and other health improvement properties have been reported. This study examined the effect of fermentation time and microbial source on physicochemical properties, phenolic compounds and antioxidant activity of two finger millet (FM) malt flours. Sorghum was used as an external reference. The grains were malted, mashed and fermented using the grain microflora and Lactobacillus fermentum. The phenolic compounds of the resulting beverage were identified and quantified using ultra-performance liquid chromatography (UPLC) and mass spectrometer system (MS). A fermentation-time dependent decrease in pH and viscosities of the beverages, with a corresponding increase in sugar content were noted. The phenolic compounds found in the FM beverages were protocatechuic acid, catechin and epicatechin. Decrease in total phenolics of the beverages was observed with increased fermentation time. The beverages exhibited 2, 2-diphenyl-1-picrylhydrazyl, 2, 2՛-azinobis-3-ethylbenzthiazoline-6-sulfonic acid radical scavenging action and iron reducing activities, which were significantly (p < 0.05) reduced at 96 h fermentation for both microbial sources. The 24 h fermented beverages retained a higher amount of total phenolics and had higher antioxidant activity compared to other fermentation periods. The study demonstrates that FM could be utilised as a functional grain in the production of non-alcoholic beverage with important phenolic compounds for health promotion and wellness.

Keywords: antioxidant activity, eleusine coracana, fermentation, phenolic compounds

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Authors: W. Stawiński, A. Wegrzyn, O. M. Freitas, S. A. Figueiredo

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Water is a life supporting resource, crucial for humanity and essential for natural ecosystems, which have been endangered by developing industry and increasing human population. Dyes are common in effluents discharged by various industries such as paper, plastics, food, cosmetics, and textile. They produce toxic effects on animals and disturb natural biological processes in receiving waters. Having complex molecular structure and resistance to biological decomposition they are problematic and difficult to be treated by conventional methods. In the search of efficient and sustainable method, sorption has been getting more interest in application to wastewaters treatment. Clays are minerals that have a layer structure based on phyllosilicate sheets that may carry a charge, which is balanced by ions located between the sheets. These charge-balancing ions can be exchanged resulting in very good ion-exchange properties of the material. Modifications of clays enhance their properties, producing a good and inexpensive sorbent for the removal of pollutants from wastewaters. The presented work proves that the treatment of a clay, vermiculite, with nitric acid followed by washing in citric acid strongly increases the sorption of two cationic dyes, methylene blue (C.I. 52015) and astrazon red (C.I. 110825). Desorption studies showed that the best eluent for regeneration is a solution of NaCl in ethanol. Cycles of sorption and desorption in column system showed no significant deterioration of sorption capacity and proved that the material shows a very good performance as sorbent, which can be recycled and reused. The results obtained open new possibilities of further modifications on vermiculite and modifications of other materials in order to get very efficient sorbents useful for wastewater treatment.

Keywords: cationic dyestuffs, sorption and regeneration, vermiculite, wastewater treatment

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Authors: Lígia Pimentel, Miguel Pereira, Manuela Pintado

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Wheat germ is a by-product of wheat flour refining. Despite this by-product being a source of proteins, lipids, fibres and complex carbohydrates, and consequently a valuable ingredient to be used in Food Industry, only few applications have been studied. The main goal of this study was to assess the potential prebiotic effect of natural wheat germ. The prebiotic potential was evaluated by in vitro assays with individual microbial strains (Lactobacillus paracasei L26 and Lactobacillus casei L431). A simulated model of the gastrointestinal digestion was also used including the conditions present in the mouth (artificial saliva), oesophagus–stomach (artificial gastric juice), duodenum (artificial intestinal juice) and ileum. The effect of natural wheat germ and wheat germ after digestion on the growth of lactic acid bacteria was studied by growing those microorganisms in de Man, Rogosa and Sharpe (MRS) broth (with 2% wheat germ and 1% wheat germ after digestion) and incubating at 37 ºC for 48 h with stirring. A negative control consisting of MRS broth without glucose was used and the substrate was also compared to a commercial prebiotic fructooligosaccharides (FOS). Samples were taken at 0, 3, 6, 9, 12, 24 and 48 h for bacterial cell counts (CFU/mL) and pH measurement. Results obtained showed that wheat germ has a stimulatory effect on the bacteria tested, presenting similar (or even higher) results to FOS, when comparing to the culture medium without glucose. This was demonstrated by the viable cell counts and also by the decrease on the medium pH. Both L. paracasei L26 and L. casei L431 could use these compounds as a substitute for glucose with an enhancement of growth. In conclusion, we have shown that wheat germ stimulate the growth of probiotic lactic acid bacteria. In order to understand if the composition of gut bacteria is altered and if wheat germ could be used as potential prebiotic, further studies including faecal fermentations should be carried out. Nevertheless, wheat germ seems to have potential to be a valuable compound to be used in Food Industry, mainly in the Bakery Industry.

Keywords: by-products, functional ingredients, prebiotic potential, wheat germ

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Authors: Jianfang Wang, Xianzhe Chen, Zhuoliang Liu, Cheng-An Tao, Yujiao Li

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Organophosphorus (OPs) pesticide used as insecticides are widely used in agricultural pest control, household and storage deworming. The detection of pesticides needs more simple and efficient methods. One of the best ways is to make electrochemical biosensors. In this paper, an electrochemical enzyme biosensor based on acetylcholine esterase (AChE) was constructed, and its sensing properties and sensing mechanisms were studied. Reduced graphene oxide-polydopamine complexes (RGO-PDA), gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) were prepared firstly and composited with AChE and chitosan (CS), then fixed on the glassy carbon electrode (GCE) surface to construct the biosensor GCE/RGO-PDA-AuNPs-AgNPs-AChE-CS by one-pot method. The results show that graphene oxide (GO) can be reduced by dopamine (DA) and dispersed well in RGO-PDA complexes. And the composites have a synergistic catalysis effect and can improve the surface resistance of GCE. The biosensor selectively can detect acetylcholine (ACh) and OPs pesticide with good linear range and high sensitivity. The performance of the biosensor is affected by the ratio and adding ways of AChE and the adding of AuNPs and AChE. And the biosensor can achieve a detection limit of 2.4 ng/L for methyl parathion and a wide linear detection range of 0.02 ng/L ~ 80 ng/L, and has excellent stability, good anti-interference ability, and excellent preservation performance, indicating that the sensor has practical value.

Keywords: acetylcholine esterase, electrochemical biosensor, nanoparticles, organophosphates, reduced graphene oxide

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Authors: K. Cartelier, D. Aime, V. Vernoud, J. Buitink, J. M. Prosperi, K. Gallardo, C. Le Signor

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Legumes can produce protein-rich seeds without nitrogen fertilizer through root symbiosis with nitrogen-fixing rhizobia. Rich in lysine, these proteins are used for human nutrition and animal feed. However, the instability of seed protein yield and quality due to environmental fluctuations limits the wider use of legumes such as pea. Breeding efforts are needed to optimize and stabilize seed nutritional value, which requires to identify the genetic determinism of seed protein plasticity in response to the environment. Towards this goal, we have studied the plasticity of protein content and composition of seeds from a collection of 200 Medicago truncatula ecotypes grown under four controlled conditions (optimal, drought, and winter/spring sowing). A quantitative analysis of one-dimensional protein profiles of these mature seeds was performed and plasticity indices were calculated from each abundant protein band. Genome-Wide Association Studies (GWAS) from these data identified major GWAS hotspots, from which a list of candidate genes was obtained. A Gene Ontology Enrichment Analysis revealed an over-representation of genes involved in several amino acid metabolic pathways. This led us to propose that environmental variations are likely to modulate amino acid balance, thus impacting seed protein composition. The selection of candidate genes for controlling the plasticity of seed protein composition was refined using transcriptomics data from developing Medicago truncatula seeds. The pea orthologs of key genes were identified for functional studies by mean of TILLING (Targeting Induced Local Lesions in Genomes) lines in this crop. We will present how this study highlighted mechanisms that could govern seed protein plasticity, providing new cues towards the stabilization of legume seed quality.

Keywords: GWAS, Medicago truncatula, plasticity, seed, storage proteins

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Authors: Daniela Ailincai, Bogdan C. Simionescu, Luminita Marin

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Polymer dispersed liquid crystals (PDLC) represent an interesting class of materials which combine the ability of polymers to form films and their mechanical strength with the opto-electronic properties of liquid crystals. The proper choice of the two components - the liquid crystal and the polymeric matrix - leads to materials suitable for a large area of applications, from electronics to biomedical devices. The objective of our work was to obtain PDLC films with potential applications in the biomedical field, using poly vinyl alcohol boric acid (PVAB) as a polymeric matrix for the first time. Presenting all the tremendous properties of poly vinyl alcohol (such as: biocompatibility, biodegradability, water solubility, good chemical stability and film forming ability), PVAB brings the advantage of containing the electron deficient boron atom, and due to this, it should promote the liquid crystal anchoring and a narrow liquid crystal droplets polydispersity. Two different PDLC systems have been obtained, by the use of two liquid crystals, a nematic commercial one: 4-cyano-4’-penthylbiphenyl (5CB) and a new smectic liquid crystal, synthesized by us: buthyl-p-[p’-n-octyloxy benzoyloxy] benzoate (BBO). The PDLC composites have been obtained by the encapsulation method, working with four different ratios between the polymeric matrix and the liquid crystal, from 60:40 to 90:10. In all cases, the composites were able to form free standing, flexible films. Polarized light microscopy, scanning electron microscopy, differential scanning calorimetry, RAMAN- spectroscopy and the contact angle measurements have been performed, in order to characterize the new composites. The new smectic liquid crystal has been characterized using 1H-NMR and single crystal X-ray diffraction and its thermotropic behavior has been established using differential scanning calorimetry and polarized light microscopy. The polarized light microscopy evidenced the formation of round birefringent droplets, anchored homeotropic in the first case and planar in the second, with a narrow dimensional polydispersity, especially for the PDLC containing the largest amount of liquid crystal, fact evidenced by SEM, also. The obtained values for the water to air contact angle showed that the composites have a proper hydrophilic-hydrophobic balance, making them potential candidates for bioapplications. More than this, our studies demonstrated that the water to air contact angle varies as a function of PVAB matrix crystalinity degree, which can be controled as a function of time. This fact allowed us to conclude that the use of PVAB as matrix for PDLCs obtaining offers the possibility to modulate their properties for specific applications.

Keywords: 4-cyano-4’-penthylbiphenyl, buthyl-p-[p’-n-octyloxy benzoyloxy] benzoate, contact angle, polymer dispersed liquid crystals, poly vinyl alcohol boric acid

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Authors: A. Merzouki, N. Haddaoui

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Polymers have a large reputation as electric insulators. These materials are characterized by weak weight, reduced price and a large domain of physical and chemical properties. They conquered new application domains that were until a recent past the exclusivity of metals. In this work, we used some composite materials (polymers/conductive fillers), as electrodes and we try to cover them with metallic lead layers in order to use them as courant collector grids in lead-acid battery plates.

Keywords: electrodeposition, polymer composites, carbon black, acetylene black

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Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate and characterize strains of Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from thirty one chicken cecal and fecal samples collected from a local farm. In vitro studies were performed to assess antibacterial activity (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), susceptibility to antibiotics (amoxycillin, cotrimoxazole, chloramphenicol, cefuroxime, ceftriaxone, ciprofloxacin, and nalidixic acid), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified using biochemical and molecular assays (API 50 CHL, and API ZYM kits followed by 16S rDNA gene sequence analysis). Two strains were identified, of which, Enteroccocus faecium was capable of inhibiting the growth of S. enteritidis and was susceptible to a wide range of antibiotics. In addition, the isolated strain exhibited significant resistance under highly acidic conditions (pH=2.5) for 8 hours and survived well in bile salt at 0.2% for 24 hours and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, the E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic

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Authors: Makhlouf Mourad, Messabih Sidi Mohamed, Bouchher Omar, Houali Farida, Benrachedi Khaled

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Mesoporous materials are very commonly used as adsorbent materials for removing phenolic compounds. However, the adsorption mechanism of these compounds is still poorly controlled. However, understanding the interactions mesoporous materials/adsorbed molecules is very important in order to optimize the processes of liquid phase adsorption. The difficulty of synthesis is to keep an orderly and cubic pore structure and achieve a homogeneous surface modification. The grafting of Si(CH₃)₃ was chosen, to transform hydrophilic surfaces hydrophobic surfaces. The aim of this work is to study the kinetics and thermodynamics of two volatile organic compounds VOC phenol (PhOH) and P hydroxy benzoic acid (4AHB) on a mesoporous material of type MCM-48 grafted with an organosilane of the Trimethylchlorosilane (TMCS) type, the material thus grafted or functionalized (hereinafter referred to as MCM-48-G). In a first step, the kinetic and thermodynamic study of the adsorption isotherms of each of the VOCs in mono-solution was carried out. In a second step, a similar study was carried out on a mixture of these two compounds. Kinetic models (pseudo-first order, pseudo-second order) were used to determine kinetic adsorption parameters. The thermodynamic parameters of the adsorption isotherms were determined by the adsorption models (Langmuir, Freundlich). The comparative study of adsorption of PhOH and 4AHB proved that MCM-48-G had a high adsorption capacity for PhOH and 4AHB; this may be related to the hydrophobicity created by the organic function of TMCS in MCM-48-G. The adsorption results for the two compounds using the Freundlich and Langmuir models show that the adsorption of 4AHB was higher than PhOH. The values ​​obtained by the adsorption thermodynamics show that the adsorption interactions for our sample with the phenol and 4AHB are of a physical nature. The adsorption of our VOCs on the MCM-48 (G) is a spontaneous and exothermic process.

Keywords: adsorption, kinetics, isotherm, mesoporous materials, Phenol, P-hydroxy benzoique acid

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Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

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Authors: Zishan Ahmad, Anwar Shahzad

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The present studies describe a protocol for high frequency in vitro propagation through nodal segments and shoot tips in D. arayalpathra, a critically endangered medicinal liana of the Western Ghats, India. Nodal segments were more responsive than shoot tips in terms of shoot multiplication. Murashige and Skoog’s (MS) basal medium supplemented with 2.5 µM 6-benzyladenine (BA) was optimum for shoot induction through both the explants. Among different combinations of plant growth regulator (PGRs) and growth additive screened, MS medium supplemented with BA (2.5 µM) + indole-3-acetic acid (IAA) (0.25 µM) + adenine sulphate (ADS) (10.0 µM) induced a maximum of 9.0 shoots per nodal segment and 3.9 shoots per shoot tip with mean shoot length of 8.5 and 3.9 cm respectively. Half-strength MS medium supplemented with Naphthaleneacetic acid (NAA) (2.5 µM) was the best for in vitro root induction. After successful acclimatization in SoilriteTM, 92 % plantlets were survived in field conditions. Acclimatized plantlets were studied for chlorophyll and carotenoid content, net photosynthetic rate (PN) and related attributes such as stomatal conductance (Gs) and transpiration rate during subsequent days of acclimatization. The rise and fall of different biochemical enzymes (SOD, CAT, APX and GR) were also studies during successful days of acclimatization. Moreover, the effect of acclimatization on the synthesis of 2-hydroxy-4-methoxy benzaldehyde (2H4MB) was also studied in relation to the biomass production. Maximum fresh weight (2.8 gm/plant), dry weight (0.35 gm/plant) of roots and 2H4MB content (8.5 µg/ ml of root extract) were recorded after 8 weeks of acclimatization. The screening of in vitro raised plantlet root was also carried out by using GC-MS analysis which witnessed more than 25 compounds. The regenerated plantlets were also screened for homogeneity by using RAPD and ISSR. The proposed protocol surely can be used for the conservation and commercial production of the plant.

Keywords: 6-benzyladenine, PGRs, RAPD, 2H4MB

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Authors: Jamilah Syafawati Yaacob, Muhammad Aiman Ramli

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Cili padi or Capsicum frutescens is one of capsicum species from nightshade family, Solanaceae. It is famous in Malaysia and is widely used as a food ingredient. Capsicum frutescens also possess vast medicinal properties. The objectives of this study are to determine the most optimum 2,4-D hormone concentration for callus induction from stem explants C. frutescens and the effects of different 2,4-D concentrations on expression of compounds from C. frutescens. Seeds were cultured on MS media without hormones (MS basal media) to yield aseptic seedlings of this species, which were then used to supply explant source for subsequent tissue culture experiments. Stem explants were excised from aseptic seedlings and cultured on MS media supplemented with various concentrations (0.1, 0.3 and 0.5 mg/L) of 2,4-D to induce formation of callus. Fresh weight, dry weight and callus growth percentage in all samples were recorded. The highest mean of dry weight was observed in MS media supplemented with 0.5 mg/L 2,4-D, where 0.4499 ± 0.106 g of callus was produced. The highest percentage of callus growth (16.4%) was also observed in cultures supplemented with 0.5 mg/L 2,4-D. The callus samples were also subjected to HPLC-MS to evaluate the effect of hormone concentration on expression of bio active compounds in different samples. Results showed that caffeoylferuloylquinic acids were present in all samples, but was most abundant in callus cells supplemented with 0.3 & 0.5 mg/L 2,4-D. Interestingly, there was an unknown compound observed to be highly expressed in callus cells supplemented with 0.1 mg/L 2,4-D, but its presence was less significant in callus cells supplemented with 0.3 and 0.5 mg/L 2,4-D. Furthermore, there was also a compound identified as octadecadienoic acid, which was uniquely expressed in callus supplemented with 0.5 mg/L 2,4-D, but absent in callus cells supplemented with 0.1 and 0.3 mg/L 2,4-D. The results obtained in this study indicated that plant growth regulators played a role in expression of secondary metabolites in plants. The increase or decrease of these growth regulators may have triggered a change in the secondary metabolite biosynthesis pathways, thus causing differential expression of compounds in this plant.

Keywords: callus, in vitro, secondary metabolite, 2, 4-Dichlorophenoxyacetic acid

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Authors: Shantanu Vyas, Neerja Meena

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Facial infiltrating lipomatosis is a rare lipomatous lesion, first described by Slavin in 1983. It is a benign pseudotumor pathology. It corresponds to a non-encapsulated collection of mature adipocytes infiltrating the local tissue and hyperplasia of underlying bone leading to a craniofacial deformity. Very few cases have been reported in the literature. We report the case of a 19-year-old female patient, who was consulted for a swelling of the right hemiface progressively evolving since birth. Physical examination revealed facial asymmetry. On palpation, the mass was soft, painless, not compressible, not pulsatile, not fluctuating. In view of the asymptomatic nature and slow progression of the lesion, a lipomatous tumour, namely lipoma, was suggested. CT scan image shows a hyperplastic subcutaneous fat on the right hemiface. On the right jugal and temporal areas, there is a subcutaneous formation of fatty density, poorly limited, with no detectable peripheral capsule. It merges with the adjacent fat. In the bone window, there was a hyperplasia of underlying bone. Facial lipomatosis infiltration of the face is a benign pseudotumor pathology. As a result, it can be confused with other disorders, in particular, hemifacial hyperplasia. Combination of physical and radiological findings can establish the diagnosis. Surgical treatment is done for cosmetic purposes.

Keywords: cosmetic correction and facial assemetry, aesthetic results, facial infiltration, surgery

Procedia PDF Downloads 76

Authors: Kairat Uteulin, Azhar Iskakova, Serik Mukhambetzhanov, Bayan Yesbolayeva, Gabit Bari, Aslan Zheksenbai, Kabyl Zhambakin, Chingis Dzhabykbayev, Vladimir Piven, Izbasar Rakhimbaiev

Abstract:

To explore the potential for in vitro rapid regeneration of Russian dandelion (Taraxacum kok-saghyz Rodin), different concentrations of 6-Benzylaminopurine (BAP), 2,4-Dichlorophenoxyacetic acid (2.4-D) and BAP combined with Indole-3-acetic acid (IAA) were evaluated for their effects on the induction of somatic embryos from leaf, seed stem and root explants. Different explants were cultured on MS medium supplemented with various concentrations (0, 0.5, 1, 1.5, 2, 2.5 and 3 mg/l) of each kind of hormone. Callus induction percentage, fresh weight, color and texture of the callus were assessed after 14 and 28 days of culture. The optimum medium for the proliferation of embryogenic calli from leaf and root explants was MS supplemented with 2.5 mg/L BAP and 0.5 mg/L 2.4-D. Concentrations of 2.5 mg/L BAP and 1.5 mg/L IAA also had a remarkable effect on root and stem explants. The best concentration to produce callus from stem explants was 0.5 mg/L BAP and 1 mg/L IAA. Results of mean comparison showed that BAP and 2.4-D were more effective on different explants than BAP and IAA. Results of the double staining method proved that somatic embryogenesis occurred in the most concentrations of BAP and 2.4-D. Under microscopic observations, the different developmental stages of the embryos (globular, heart, torpedo and cotyledonary) were revealed together in callus cells, indicating that the most tested hormone combinations were effective for somatic embryogenesis formation in this species. Seed explants formed torpedo and cotyledonary stages faster than leaf and root explants in the most combinations. Most calli from seed explants were cream colored and friable, while calli were compact and light green from leaf and root explants. Some combinations gave direct regeneration and (3 mg/L BAP and 2 mg/L IAA) in seed explants and (0.5 mg/L BAP and 2.5 mg/L IAA) in leaf explants had the highest number of shoots with average of 21 and 27 shoots per callus. The developed protocol established the production of different callus types from seed, leaf, and root explants and plant regeneration through somatic embryogenesis.

Keywords: taraxacum kok-saghyz Rodin, callus, somatic embryogenesis

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Authors: Mohd Ali Abbas Zaidi, Meenu Sachdeva, Jonaki Sen

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In the vertebrate embryo, the forebrain anlagen develops from the anterior-most region of the neural tube which is the precursor of the central nervous system (CNS). The roof plate located at the dorsal midline region of the forebrain anlagen, acts as a source of several secreted molecules involved in patterning and morphogenesis of the forebrain. One such key morphogenetic event is the invagination of the forebrain roof plate which results in separation of the single forebrain vesicle into two cerebral hemispheres. Retinoic acid (RA) signaling plays a key role in this process. Blocking RA signaling at the dorsal forebrain midline inhibits dorsal invagination and results in the absence of certain key features of this region, such as thinning of the neuroepithelium and a lowering of cell proliferation. At present we are investigating the possibility of other signaling pathways acting in concert with RA signaling to regulate this process. We have focused on BMP signaling, which we found to be active in a mutually exclusive domain to that of RA signaling within the roof plate. We have also observed that there is a change in BMP signaling activity on modulation of RA signaling indicating an antagonistic relationship between the two. Moreover, constitutive activation of BMP signaling seems to completely inhibit thinning and partially affect invagination, leaving the lowering of cell proliferation in the midline unaffected. We are employing in-silico modeling as well as molecular manipulations to investigate the relative contribution if any, of regional differences in rates of cell proliferation and thinning of the neuroepithelium towards the process of invagination. We have found expression of certain cell adhesion molecules in forebrain roof-plate whose mRNA localization across the thickness of neuroepithelium is influenced by Bmp and RA signaling, giving regional rigidity to roof plate and assisting invagination. We also found expression of certain cytoskeleton modifiers in a localized small domains in invaginating forebrain roof plate suggesting that midline invagination is under control of many factors.

Keywords: bone morphogenetic signaling, cytoskeleton, cell adhesion molecules, forebrain roof plate, retinoic acid signaling

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Authors: Claudia Schier, Arvid Biallas, Bernd Friedrich

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The demand for Li-ion-batteries owing to their benefits, such as; fast charging time, high energy density, low weight, large temperature range, and a long service life performance is increasing compared to other battery systems. These characteristics are substantial not only for battery-operated portable devices but also in the growing field of electromobility where high-performance energy storage systems in the form of batteries are highly requested. Due to the sharp rising production, there is a tremendous interest to recycle spent Li-Ion batteries in a closed-loop manner owed to the high content of valuable metals such as cobalt, manganese, and lithium as well as regarding the increasing demand for those scarce applied metals. Currently, there are just a few industrial processes using hydrometallurgical methods to recover valuable metals from Li-ion-battery waste. In this study, the extraction of valuable metals from spent Li-ion-batteries is investigated by pretreated and subsequently leached battery wastes using different precipitation methods in a comparative manner. For the extraction of lithium, cobalt, and other valuable metals, pelletized battery wastes with an initial Li content of 2.24 wt. % and cobalt of 22 wt. % is used. Hydrochloric acid with 4 mol/L is applied with 1:50 solid to liquid (s/l) ratio to generate pregnant leach solution for subsequent precipitation steps. In order to obtain pure precipitates, two different pathways (pathway 1 and pathway 2) are investigated, which differ from each other with regard to the precipitation steps carried out. While lithium carbonate recovery is the final process step in pathway 1, pathway 2 requires a preliminary removal of lithium from the process. The aim is to evaluate both processes in terms of purity and yield of the products obtained. ICP-OES is used to determine the chemical content of leach liquor as well as of the solid residue.

Keywords: hydrochloric acid, hydrometallurgy, Li-ion-batteries, metal recovery

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Authors: Annisa Ulfah Pristya, Andi Setiawan

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Indonesia placed in sixth rank total Acute Respiratory Infection (ARI) patient in the world and Riau as one of the province with the highest number of people with respiratory infection in Indonesia reached 37 thousand people. Usually society using a mask as prevention action. Unfortunately the commercial mouth mask only can work maximum for 4 hours and the pores are too large to filter out microorganisms and viruses carried by infectious droplets nucleated 1-5 μm. On the other hand, Indonesia is rich with Titanium dioxide (TiO2) and purple sweet potato anthocyanin pigment. Therefore, offered Bio-nano-mask which is a antimicrobial and antiviral mouth mask with Nano-TiO2 coating and purple sweet potato anthocyanins utilization as an effective solution to high ARI patients in Riau, which has the advantage of the mask surface can’t be attached by infectious droplets, self-cleaning and have anthocyanins biosensors that give visual response can be understood easily by the general public in the form of a mask color change from blue/purple to pink when acid levels increase. Acid level is an indicator of microorganisms accumulation in the mouth and surrounding areas. Bio-nano mask making process begins with the preparation (design, Nano-TiO2 liquid preparation, anthocyanins biosensors manufacture) and then superimposing the Nano-TiO2 on the outer surface of spunbond color using a sprayer, then superimposing anthocyanins biosensors film on the Meltdown surface, making bio nano-mask and it pack. Bio-nano mask has the advantage is effectively preventing pathogenic microorganisms and infectious droplets and has accumulated indicator microorganisms that color changes which easily observed by the common people though.

Keywords: anthocyanins, ARI, nano-TiO2 liquid, self cleaning

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Authors: N. A. Hamizi, M. R. Johan, Y. H. Hor, A. N. Sabri, Y. Y. A. Yong

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In this research, Mn-doped CdSe QDs is synthesized by using paraffin liquid as the reacting solvent and oleic acid as the ligands for Cd in order to produce Mn-doped CdSe QDs in zinc-blende crystal structure. Characterization studies for synthesized Mn-doped CdSe QDs are carried out using UV-visible and photoluminescence spectroscopy. The absorption wavelengths in UV-vis test and emission wavelengths in PL test were increase with the increases in the ripening temperature and time respectively.

Keywords: semiconductor, chemical synthesis, optical properties, ripening

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Authors: D: Gupta, M. Radhakrishnan, Y. Kurhe, D. Thangaraj

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Depression is among the leading causes of death worldwide. The current pharmacotherapy is associated with poor compliance, resistance and relapse, which necessitate the development of novel compounds with better efficacy. The present study designed and synthesized EQC-34 (N-cyclohexyl-3-ethoxyquinoxalin-2-carboxamide) as novel serotonin type-3 (5HT3) antagonist and evaluated its antidepressant-like effects using neurobehavioral mouse model. 5HT3 antagonism (as pA2 value) was determined on the longitudinal smooth muscle of guinea-pig ileum against 2-methyl-5HT (a 5HT3 agonist). The doses were calculated by dose response of basal locomotor activity. Consequently, effects of EQC-34 on neurobehavioral parameters were measured in forced swim (FST) and tail suspension test (TST). The possible mechanism was estimated by interaction study with fluoxetine (a selective serotonin reuptake inhibitor) and mCPBG (1-(m-chlorophenyl)-biguanide, a selective 5HT3 agonist), and confirmed by potentiation of head twitch response by 5hydroxy-L-tryptophan (5HTP). EQC-34 (1-4 mg/kg, i.p.) produced significant decreased behavioral despair effects in FST and TST. It potentiated fluoxetine response, while mCPBG reduced EQC-34 activity in FST. Further, EQC-34 potentiated 5HTP induced head twitch response. EQC-34 revealed potential antidepressant-like effects, which may involve 5HT3 receptor mediated facilitation of 5HT neurotransmission, thereby reversing the pathological deficiency of monoamines (5HT) observed in depression. Thus, it may be further investigated as promising agent to improve therapeutics of depression.

Keywords: depression, forced swim test, 5HT3 receptor antagonist, serotonin

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Authors: Afshin Shahbazi, Hadi Shadi Naghadeh, Ahmad Khodadadi Darban

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In the Magnetically Assisted Chemical Separation (MACS) process, tiny ferromagnetic particles coated with solvent extractant are used to selectively separate radionuclides and hazardous metals from aqueous waste streams. The contaminant-loaded particles are then recovered from the waste solutions using a magnetic field. In the present study, Cyanex272 or C272 (bis (2,4,4-trimethylpentyl) phosphinic acid) coated magnetic particles are being evaluated for the possible application in the extraction of Thorium (IV) from nuclear waste streams. The uptake behaviour of Th(IV) from nitric acid solutions was investigated by batch studies. Adsorption of Thorium (IV) from aqueous solution onto adsorbent was investigated in a batch system. Adsorption isotherm and adsorption kinetic studies of Thorium (IV) onto nanoparticles coated Cyanex272 were carried out in a batch system. The factors influencing Thorium (IV) adsorption were investigated and described in detail, as a function of the parameters such as initial pH value, contact time, adsorbent mass, and initial Thorium (IV) concentration. Magnetically Assisted Chemical Separation (MACS) process adsorbent showed best results for the fast adsorption of Th (IV) from aqueous solution at aqueous phase acidity value of 0.5 molar. In addition, more than 80% of Th (IV) was removed within the first 2 hours, and the time required to achieve the adsorption equilibrium was only 140 minutes. Langmuir and Frendlich adsorption models were used for the mathematical description of the adsorption equilibrium. Equilibrium data agreed very well with the Langmuir model, with a maximum adsorption capacity of 48 mg.g-1. Adsorption kinetics data were tested using pseudo-first-order, pseudo-second-order and intra-particle diffusion models. Kinetic studies showed that the adsorption followed a pseudo-second-order kinetic model, indicating that the chemical adsorption was the rate-limiting step.

Keywords: Thorium (IV) adsorption, MACS process, magnetic nanoparticles, Cyanex272

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Authors: Lin Meng, Xi Lu, Haibo Wang, Yong Song, Lili Cao, Wenfang Song, Yong Hu

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China has abundant heavy oil resources, and thermal recovery is the main recovery method for heavy oil reservoirs. However, high energy consumption, high carbon emission and high production costs make heavy oil thermal recovery unsustainable. It is urgent to explore a replacement for developing technology. A low Carbon and cost technology of heavy oil recovery, chemical viscosity-reduction in layer (CVRL), is developed by the petroleum exploration and development research institute of Sinopec via investigated mechanisms, synthesized products, and improved oil production technologies, as follows: (1) Proposed a cascade viscous mechanism of heavy oil. Asphaltene and resin grow from free molecules to associative structures further to bulk aggregations by π - π stacking and hydrogen bonding, which causes the high viscosity of heavy oil. (2) Aimed at breaking the π - π stacking and hydrogen bond of heavy oil, the copolymer of N-(3,4-dihydroxyphenethyl) acryl amide and 2-Acrylamido-2-methylpropane sulfonic acid was synthesized as a viscosity reducer. It achieves a viscosity reduction rate of>80% without shearing for heavy oil (viscosity < 50000 mPa‧s), of which fluidity is evidently improved in the layer. (3) Synthesized hydroxymethyl acrylamide-maleic acid-decanol ternary copolymer self-assembly plugging agent. The particle size is 0.1 μm-2 mm adjustable, and the volume is 10-500 times controllable, which can achieve the efficient transportation of viscosity reducer to enriched oil areas. CVRL has applied 400 wells until now, increasing oil production by 470000 tons, saving 81000 tons of standard coal, reducing CO2 emissions by 174000 tons, and reducing production costs by 60%. It promotes the transformation of heavy oil towards low energy consumption, low carbon emissions, and low-cost development.

Keywords: heavy oil, chemical viscosity-reduction, low carbon, viscosity reducer, plugging agent

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Authors: Alexander A. Tokmakov

Abstract:

Cell-free protein synthesis is widely used to synthesize recombinant proteins. It allows genome-scale expression of various polypeptides under strictly controlled uniform conditions. However, only a minor fraction of all proteins can be successfully expressed in the systems of protein synthesis that are currently used. The factors determining expression success are poorly understood. At present, the vast volume of data is accumulated in cell-free expression databases. It makes possible comprehensive bioinformatics analysis and identification of multiple features associated with successful cell-free expression. Here, we describe an approach aimed at identification of multiple physicochemical and structural properties of amino acid sequences associated with protein solubility and aggregation and highlight major correlations obtained using this approach. The developed method includes: categorical assessment of the protein expression data, calculation and prediction of multiple properties of expressed amino acid sequences, correlation of the individual properties with the expression scores, and evaluation of statistical significance of the observed correlations. Using this approach, we revealed a number of statistically significant correlations between calculated and predicted features of protein sequences and their amenability to cell-free expression. It was found that some of the features, such as protein pI, hydrophobicity, presence of signal sequences, etc., are mostly related to protein solubility, whereas the others, such as protein length, number of disulfide bonds, content of secondary structure, etc., affect mainly the expression propensity. We also demonstrated that amenability of polypeptide sequences to cell-free expression correlates with the presence of multiple sites of post-translational modifications. The correlations revealed in this study provide a plethora of important insights into protein folding and rationalization of protein production. The developed bioinformatics approach can be of practical use for predicting expression success and optimizing cell-free protein synthesis.

Keywords: bioinformatics analysis, cell-free protein synthesis, expression success, optimization, recombinant proteins

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Authors: Karim Hrratha, Khaled Essalahb, Christophe Morellc, Henry Chermettec, Salima Boughdiria

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Among the carbon-carbon bond formation types, allylation of active methylene compounds with cyclic Baylis-Hillman (BH) alcohols is a reliable and widely used method. This reaction is a very attractive tool in organic synthesis of biological and biodiesel compounds. Thus, in view of an insistent and peremptory request for an efficient and straightly method for synthesizing the desired product, a thorough analysis of various aspects of the reaction processes is an important task. The product afforded by the reaction of active methylene with BH alcohols depends largely on the experimental conditions, notably on the catalyst properties. All experiments reported that catalysis is needed for this reaction type because of the poor ability of alcohol hydroxyl group to be as a suitable leaving group. Within the catalysts, several transition- metal based have been used such as palladium in the presence of acid or base and have been considered as reliable methods. Furthemore, acid catalysts such as BF3.OEt2, BiX3 (X= Cl, Br, I, (OTf)3), InCl3, Yb(OTf)3, FeCl3, p-TsOH and H-montmorillonite have been employed to activate the C-C bond formation through the alkylation of active methylene compounds. Interestingly a report of a smoothly process for the ability of 4-imethyaminopyridine(DMAP) to catalyze the allylation reaction of active methylene compounds with cyclic Baylis-Hillman (BH) alcohol appeared recently. However, the reaction mechanism remains ambiguous, since the C- allylation process leads to an unexpected product (noted P1), corresponding to a direct allylation instead of conjugate allylation, which involves the most electrophilic center according to the electron withdrawing group CO effect. The main objective of the present theoretical study is to better understand the role of the DMAP catalytic activity as well as the process leading to the end- product (P1) for the catalytic reaction of a cyclic BH alcohol with active methylene compounds. For that purpose, we have carried out computations of a set of active methylene compounds varying by R1 and R2 toward the same alcohol, and we have attempted to rationalize the mechanisms thanks to the acid–base approach, and conceptual DFT tools such as chemical potential, hardness, Fukui functions, electrophilicity index and dual descriptor, as these approaches have shown a good prediction of reactions products.The present work is then organized as follows: In a first part some computational details will be given, introducing the reactivity indexes used in the present work, then Section 3 is dedicated to the discussion of the prediction of the selectivity and regioselectivity. The paper ends with some concluding remarks. In this work, we have shown, through DFT method at the B3LYP/6-311++G(d,p) level of theory that: The allylation of active methylene compounds with cyclic BH alcohol is governed by orbital control character. Hence the end- product denoted P1 is generated by direct allylation.

Keywords: DFT calculation, gas phase pKa, theoretical mechanism, orbital control, charge control, Fukui function, transition state

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