Search results for: caffeic acid analysis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 29605

Search results for: caffeic acid analysis

28735 Eu³⁺ PVC Membrane Sensor Based on 1,2-Diaminopropane-N,N,N',N'-Tetraacetic Acid

Authors: Noshin Mehrabian, Mohammad Reza Abedi, Hassan Ali Zamani

Abstract:

A highly selective poly(vinyl chloride)-based membrane sensor produced by using 1,2-Diaminopropane-N,N,N',N'-tetraacetic acid (DAPTA) as active material is described. The electrode displays Nernstian behavior over the concentration range 1.0×10⁻⁶ to 1.0×10⁻² M. The detection limit of the electrode is 7.2×10⁻⁷ M. The best performance was obtained with the membrane containing 30% polyvinyl chloride (PVC), 65% nitrobenzene (NB), 2% sodium tetra phenyl borate (Na TPB), 3% DAPTA. The potentiometric response of the proposed electrode is pH independent in the range of 2.5–‎‎9.1. ‎The proposed sensor displays a fast response time 'less than 10s'. The electrode shows a good selectivity for Eu (III) ion with respect to most common cations including alkali, alkaline earth, transition, and heavy metal ions. It was used as an indicator electrode in potentiometric ‎titration of 25 mL of a 1.0×10⁻⁴ M Eu (III) solution with a 1.0×10⁻² M EDTA solution.

Keywords: potentiometry, PVC membrane, sensor, ion-selective electrode

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28734 Geochemistry of Natural Radionuclides Associated with Acid Mine Drainage (AMD) in a Coal Mining Area in Southern Brazil

Authors: Juliana A. Galhardi, Daniel M. Bonotto

Abstract:

Coal is an important non-renewable energy source of and can be associated with radioactive elements. In Figueira city, Paraná state, Brazil, it was recorded high uranium activity near the coal mine that supplies a local thermoelectric power plant. In this context, the radon activity (Rn-222, produced by the Ra-226 decay in the U-238 natural series) was evaluated in groundwater, river water and effluents produced from the acid mine drainage in the coal reject dumps. The samples were collected in August 2013 and in February 2014 and analyzed at LABIDRO (Laboratory of Isotope and Hydrochemistry), UNESP, Rio Claro city, Brazil, using an alpha spectrometer (AlphaGuard) adjusted to evaluate the mean radon activity concentration in five cycles of 10 minutes. No radon activity concentration above 100 Bq.L-1, which was a previous critic value established by the World Health Organization. The average radon activity concentration in groundwater was higher than in surface water and in effluent samples, possibly due to the accumulation of uranium and radium in the aquifer layers that favors the radon trapping. The lower value in the river waters can indicate dilution and the intermediate value in the effluents may indicate radon absorption in the coal particles of the reject dumps. The results also indicate that the radon activities in the effluents increase with the sample acidification, possibly due to the higher radium leaching and the subsequent radon transport to the drainage flow. The water samples of Laranjinha River and Ribeirão das Pedras stream, which, respectively, supply Figueira city and receive the mining effluent, exhibited higher pH values upstream the mine, reflecting the acid mine drainage discharge. The radionuclides transport indicates the importance of monitoring their activity concentration in natural waters due to the risks that the radioactivity can represent to human health.

Keywords: radon, radium, acid mine drainage, coal

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28733 Miniaturizing the Volumetric Titration of Free Nitric Acid in U(vi) Solutions: On the Lookout for a More Sustainable Process Radioanalytical Chemistry through Titration-On-A-Chip

Authors: Jose Neri, Fabrice Canto, Alastair Magnaldo, Laurent Guillerme, Vincent Dugas

Abstract:

A miniaturized and automated approach for the volumetric titration of free nitric acid in U(VI) solutions is presented. Free acidity measurement refers to the acidity quantification in solutions containing hydrolysable heavy metal ions such as U(VI), U(IV) or Pu(IV) without taking into account the acidity contribution from the hydrolysis of such metal ions. It is, in fact, an operation having an essential role for the control of the nuclear fuel recycling process. The main objective behind the technical optimization of the actual ‘beaker’ method was to reduce the amount of radioactive substance to be handled by the laboratory personnel, to ease the instrumentation adjustability within a glove-box environment and to allow a high-throughput analysis for conducting more cost-effective operations. The measurement technique is based on the concept of the Taylor-Aris dispersion in order to create inside of a 200 μm x 5cm circular cylindrical micro-channel a linear concentration gradient in less than a second. The proposed analytical methodology relies on the actinide complexation using pH 5.6 sodium oxalate solution and subsequent alkalimetric titration of nitric acid with sodium hydroxide. The titration process is followed with a CCD camera for fluorescence detection; the neutralization boundary can be visualized in a detection range of 500nm- 600nm thanks to the addition of a pH sensitive fluorophore. The operating principle of the developed device allows the active generation of linear concentration gradients using a single cylindrical micro channel. This feature simplifies the fabrication and ease of use of the micro device, as it does not need a complex micro channel network or passive mixers to generate the chemical gradient. Moreover, since the linear gradient is determined by the liquid reagents input pressure, its generation can be fully achieved in faster intervals than one second, being a more timely-efficient gradient generation process compared to other source-sink passive diffusion devices. The resulting linear gradient generator device was therefore adapted to perform for the first time, a volumetric titration on a chip where the amount of reagents used is fixed to the total volume of the micro channel, avoiding an important waste generation like in other flow-based titration techniques. The associated analytical method is automated and its linearity has been proven for the free acidity determination of U(VI) samples containing up to 0.5M of actinide ion and nitric acid in a concentration range of 0.5M to 3M. In addition to automation, the developed analytical methodology and technique greatly improves the standard off-line oxalate complexation and alkalimetric titration method by reducing a thousand fold the required sample volume, forty times the nuclear waste per analysis as well as the analysis time by eight-fold. The developed device represents, therefore, a great step towards an easy-to-handle nuclear-related application, which in the short term could be used to improve laboratory safety as much as to reduce the environmental impact of the radioanalytical chain.

Keywords: free acidity, lab-on-a-chip, linear concentration gradient, Taylor-Aris dispersion, volumetric titration

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28732 Evaluation of Real Time PCR Methods for Food Safety

Authors: Ergun Sakalar, Kubra Bilgic

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In the last decades, real-time PCR has become a reliable tool preferred to use in many laboratories for pathogen detection. This technique allows for monitoring target amplification via fluorescent molecules besides admit of quantitative analysis by enabling of convert outcomes of thermal cycling to digital data. Sensitivity and traceability of real-time PCR are based on measuring of fluorescence that appears only when fluorescent reporter dye bound to specific target DNA.The fluorescent reporter systems developed for this purpose are divided into two groups. The first group consists of intercalator fluorescence dyes such as SYBR Green, EvaGreen which binds to double-stranded DNA. On the other hand, the second group includes fluorophore-labeled oligonucleotide probes that are separated into three subgroups due to differences in mechanism of action; initial primer-probes such as Cyclicons, Angler®, Amplifluor®, LUX™, Scorpions, and the second one hydrolysis probes like TaqMan, Snake assay, finally hybridization probes, for instance, Molecular Beacons, Hybprobe/FRET, HyBeacon™, MGB-Eclipse, ResonSense®, Yin-Yang, MGB-Pleiades. In addition nucleic acid analogues, an increase of probe affinity to target site is also employed with fluorescence-labeled probes. Consequently, abundant real-time PCR detection chemistries are chosen by researcher according to the field of application, mechanism of action, advantages, and proper structures of primer/probes.

Keywords: fluorescent dye, food safety, molecular probes, nucleic acid analogues

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28731 Investigation of Fumaric Acid Radiolysis Using Gamma Irradiation

Authors: Wafa Jahouach-Rabai, Khouloud Ouerghi, Zohra Azzouz-Berriche, Faouzi Hosni

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Widely used organic products in the pharmaceutical industry have been detected in environmental systems, essentially carboxylic acids. In this purpose, the degradation efficiency of these contaminants was evaluated using an advanced oxidation process (AOP), namely ionization process as an alternative to conventional water treatment technologies. This process permitted the generation of radical reactions to directly degrade organic pollutants in wastewater. In fact, gamma irradiation of aqueous solutions produces several reactive radicals, essentially hydroxyl radical (OH), to destroy recalcitrant pollutants. Different concentrations of aqueous solutions of Fumaric acid (FA) were considered in this study (0.1-1 mmol/L), which were treated by irradiation doses from 1 to 15 kGy with 6.1 kGy/h rate by ionizing system in pilot scale (⁶⁰Co irradiator). Variations of main parameters influencing degradation efficiency versus absorbed doses were released in the aim to optimize total mineralization of considered pollutants. Preliminary degradation pathway until complete mineralization into CO₂ has been suggested based on detection of residual degradation derivatives using different techniques, namely high performance liquid chromatography (HPLC) and electron paramagnetic resonance spectroscopy (EPR). Results revealed total destruction of treated compound, which improve the efficiency of this process in water remediation. We investigated the reactivity of hydroxyl radicals generated by irradiation on dicarboxylic acid (FA) in aqueous solutions, leading to its degradation into other smaller molecules. In fact, gamma irradiation of FA leads to the formation of hydroxylated intermediates such as hydroxycarbonyl radical which were identified by EPR spectroscopy. Finally, pilot plant irradiation facilities improved the applicability of radiation technology on large scale.

Keywords: AOP, radiolysis, fumaric acid, gamma irradiation, hydroxyl radical, EPR, HPLC

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28730 The Effect of Gamma-Aminobutyric Acid on Mechanical Properties, Water Vapor Permeability and Solubility of Pectin Films

Authors: Jitrawadee Meerasri, Rungsinee Sothornvit

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Pectin is a structural polysaccharide from plant cell walls and can be used as a stabilizer, gelling and film-forming agents to improve many food products. Moreover, pectin film as a natural biopolymer can be a carrier of several active ingredients such as antioxidant and antimicrobial to provide an active or functional film. Gamma-aminobutyric acid (GABA) is a well-known agent to reduce neuronal excitability throughout the nervous system and it is interesting to investigate the GABA effect as a substitute of normal plasticizer (glycerol) on edible film properties. Therefore, the objective of this study was to determine the effect of GABA concentrations (5-15% of pectin) on film mechanical properties, moisture content, water vapor permeability, and solubility compared with those from glycerol (10% of pectin) plasticized pectin film including a control film (pectin film without any plasticizer). It was found that an increase in GABA concentrations decreased film tensile strength, modulus, solubility and water vapor permeability, but elongation was increased without a change in the moisture content. The smaller amount of GABA showed the equivalent film properties as using a higher amount of glycerol. Consequently, GABA can act as an alternative plasticizer substitute of glycerol at the lower amount used. Moreover, GABA provides the nutritional high value in the food products when the edible packaging material is consumed with products.

Keywords: gamma-aminobutyric acid, pectin, plasticizer, edible film

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28729 Zebrafish Larvae Model: A High Throughput Screening Tool to Study Autism

Authors: Shubham Dwivedi, Raghavender Medishetti, Rita Rani, Aarti Sevilimedu, Pushkar Kulkarni, Yogeeswari Perumal

Abstract:

Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder of early onset, characterized by impaired sociability, cognitive function and stereotypies. There is a significant urge to develop and establish new animal models with ASD-like characteristics for better understanding of underlying mechanisms. The aim of the present study was to develop a cost and time effective zebrafish model with quantifiable parameters to facilitate mechanistic studies as well as high-throughput screening of new molecules for autism. Zebrafish embryos were treated with valproic acid and a battery of behavioral tests (anxiety, inattentive behavior, irritability and social impairment) was performed on larvae at 7th day post fertilization, followed by study of molecular markers of autism. This model shows a significant behavioural impairment in valproic acid treated larvae in comparison to control which was again supported by alteration in few marker genes and proteins of autism. The model also shows a rescue of behavioural despair with positive control drugs. The model shows robust parameters to study behavior, molecular mechanism and drug screening approach in a single frame. Thus we postulate that our 7 days zebrafish larval model for autism can help in high throughput screening of new molecules on autism.

Keywords: autism, zebrafish, valproic acid, neurodevelopment, behavioral assay

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28728 Determination of Physicochemical Properties, Bioaccessibility of Phenolics and Antioxidant Capacity of Mineral Enriched Linden Herbal Tea Beverage

Authors: Senem Suna, Canan Ece Tamer, Ömer Utku Çopur

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In this research, dried linden (Tilia argentea) leaves and blossoms were used as a raw material for mineral enriched herbal tea beverage production. For this aim, %1 dried linden was infused with boiling water (100 °C) for 5 minutes. After cooling, sucrose, citric acid, ascorbic acid, natural lemon flavor and natural mineral water were added. Beverage samples were plate filtered, filled into 200-mL glass bottles, capped then pasteurized at 98 °C for 15 minutes. Water soluble dry matter, titratable acidity, ascorbic acid, pH, minerals (Fe, Ca, Mg, K, Na), color (L*, a*, b*), turbidity, bioaccessible phenolics and antioxidant capacity were analyzed. Water soluble dry matter, titratable acidity, and ascorbic were determined as 7.66±0.28 g/100 g, 0.13±0.00 g/100 mL, and 19.42±0.62 mg/100 mL, respectively. pH was measured as 3.69. Fe, Ca, Mg, K and Na contents of the beverage were determined as 0.12±0.00, 115.48±0.05, 34.72±0.14, 48.67±0.43 and 85.72±1.01 mg/L, respectively. Color was measured as 13.63±0.05, -4.33±0.05, and 3.06±0.05 for L*, a*, and b* values. Turbidity was determined as 0.69±0.07 NTU. Bioaccessible phenolics were determined as 312.82±5.91 mg GAE/100 mL. Antioxidant capacities of chemical (MetOH:H2O:HCl) and physiological extracts (in vitro digestive enzymatic extraction) with DPPH (27.59±0.53 and 0.17±0.02 μmol trolox/mL), FRAP (21.01±0.97 and 13.27±0.19 μmol trolox/mL) and CUPRAC (44.71±9.42 and 2.80±0.64 μmol trolox/mL) methods were also evaluated. As a result, enrichment with natural mineral water was proposed for the development of functional and nutritional values together with a good potential for commercialization.

Keywords: linden, herbal tea beverage, bioaccessibility, antioxidant capacity

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28727 Use of Pig as an Animal Model for Assessing the Differential MicroRNA Profiling in Kidney after Aristolochic Acid Intoxication

Authors: Daniela E. Marin, Cornelia Braicu, Gina C. Pistol, Roxana Cojocneanu-Petric, Ioana Berindan Neagoe, Mihail A. Gras, Ionelia Taranu

Abstract:

Aristolochic acid (AA) is a carcinogenic, mutagenic, and nephrotoxic compound commonly found in the Aristolochiaceae family of plants. AA is frequently associated with urothelial carcinoma of the upper urinary tract in human and animals and is considered as being responsible for Balkan Endemic Nephropathy. The pig provides a good animal model because the porcine urological system is very similar to that of humans, both in aspects of physiology and anatomy. MicroRNA (miRNA) are small non-coding RNAs that have an impact on a wide range of biological processes by regulating gene expression at post-transcriptional level. The objective of this study was to analyze the miRNA profiling in the kidneys of AA intoxicated swine. For this purpose, ten TOPIGS-40 crossbred weaned piglets, 4-week-old, male and females with an initial average body weight of 9.83 ± 0.5 kg were studied for 28 days. They were given ad libitum access to water and feed and randomly allotted to one of the following groups: control group (C) or aristolochic acid group (AA). They were fed a maize-soybean-meal-based diet contaminated or not with 0.25mgAA/kg. To profile miRNA in the kidneys of pigs, microarrays and bioinformatics approaches were applied to analyze the miRNA in the kidney of control and AA intoxicated pigs. After normalization, our results have shown that a total of 5 known miRNAs and 4 novel miRNAs had different profiling in the kidney of intoxicated animals versus control ones. Expression of miR-32-5p, miR-497-5p, miR-423-3p, miR-218-5p, miR-128-3p were up-regulated by 0.25mgAA/kg feed, while the expression of miR-9793-5p, miR-9835-3p, miR-9840-3p, miR-4334-5p was down-regulated. The microRNA profiling in kidney of intoxicated animals was associated with modified expression of target genes as: RICTOR, LASP1, SFRP2, DKK2, BMI1, RAF1, IGF1R, MAP2K1, WEE1, HDGF, BCL2, EIF4E etc, involved in cell division cycle, apoptosis, cell differentiation and cell migration, cell signaling, cancer etc. In conclusion, this study provides new data concerning the microRNA profiling in kidney after aristolochic acid intoxications with important implications for human and animal health.

Keywords: aristolochic acid, kidney, microRNA, swine

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28726 Quantitative Analysis of the High-Value Bioactive Components of Pre-Germinated and Germinated Pigmented Rice (Oryza sativa L. Cv. Superjami and Superhongmi)

Authors: Lara Marie Pangan Lo, Soo Im Chung, Yao Cheng Zhang, Xingyue Jin, Mi Young Kang

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Being the world’s most consumed grain crop, rice (Oryza sativa L.) demands’ have increase and this prompted the development of new rice cultivars with high bio-functional properties than the commonly used white rice. Ordinary rice variety is already known to be a potential source for a number of nutritional as well as bioactive compounds. To further enhance the rice’s nutritive values, germination is done in addition to making it more tasty and palatable when cooked. Pigmented rice, on the other hand, has become increasingly popular in the recent years for their greater antioxidant potential and other nutraceutical properties which can help alleviate the occurrence of the increasing incidence of metabolic diseases. Combining these two (2) parameters, this research study is sought to quantitatively determine the pre-germinated and germinated quantities of the major bioactive compounds of South Korea’s newly developed purplish pigmented rice grain cultivar Superjami (SJ) and red pigmented rice grain Superhongmi (SH) and compare them against the non-pigmented Normal Brown (NB) rice variety. Powdered rice grain cultivars were subjected to 72-hour germination period and the quantities of GABA, γ-oryzanol, ferulic acid, tocopherol and tocotrienol homologues were compared against their pre-germinated condition using γ- amino butyric acid (GABA) analysis and High Performance Liquid Chromatography (HPLC). Results revealed the effectiveness of germination in enhancing the bioactive components in all rice samples. GABA contents in germinated rice cultivars increased by more than 10-fold following the order: SJ >SH >NB. In addition, purple rice variety (SJ) has higher total γ-oryzanol and ferulic acid contents which increased by > 2-fold after germination followed by the red cultivar SH then the control, NB. Germinated varieties also possess higher total tocotrienol content than their pre-germinated state. As for the total tocopherol content, SJ has higher quantity, but the red-pigmented SH (0.16 mg/kg) is shown to have lower total tocopherol content than the control rice NB (0.86 mg/kg). However, all tocopherol and tocotrienol homologues were present only in small amounts ( < 3.0 mg/kg) in all pre-germinated and germinated samples. In general, all of the analyzed pigmented rice cultivars were found to possess higher bioactive compounds than the control NB rice variety. Also, regardless of their strain, germinated rice samples have higher bioactive compounds than their pre-germinated counterparts. This only shows the effectiveness of germinating rice in enhancing bioactive constituents. Overall, these results suggest the potential of the pigmented rice varieties as natural source of nutraceuticals in bio-functional food development.

Keywords: bioactive compounds, germinated rice, superhongmi, superjami

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28725 Influence of Hydrolytic Degradation on Properties of Moisture Membranes Used in Fire-Protective Clothing

Authors: Rachid El Aidani, Phuong Nguyen-Tri, Toan Vu-Khanh

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This study intends to show the influence of the hydrolytic degradation on the properties of the e-PTFE/NOMEX® membranes used in fire-protective clothing. The modification of water vapour permeability, morphology and chemical structure was examined by MOCON Permatran, electron microscopy scanning (SEM), and ATR-FTIR, respectively. A decrease in permeability to water vapour of the aged samples was observed following closure of transpiration pores. Analysis of fiber morphology indicates the appearance of defects at the fibers surface with the presence of micro cavities as well as the of fibrils. ATR-FTIR analysis reveals the presence of a new absorption band attributed to carboxylic acid terminal groups generated during the amide bond hydrolysis.

Keywords: hydrolytic ageing, moisture membrane, water vapor permeability, morphology

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28724 Functionalized PU Foam for Water Filtration

Authors: Nidal H. Abu-Zahra, Subhashini Gunashekar

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Polyurethane foam is functionalized with Sulfonic acid groups to remove lead ions (Pb2+) from drinking water through a action exchange process. The synthesis is based on addition polymerization of the -NCO groups of an isocyanine with the –OH groups of a polio to form the urethane. Toluene-diisocyanateis reacted with Polypropylene glycol to form a linear pre-polymer, which is further polymerized using a chain extender, N, N-bis(2-hydorxyethyl)-2-aminoethane-sulfonic acid (BES). BES acts as a functional group site to exchange Pb2+ ions. A set of experiments was designed to study the effect of various processing parameters on the performance of the synthesized foam. The maximum Pb2+ ion exchange capacity of the foam was found to be 47ppb/g from a 100ppb Pb2+ solution over a period of 60 minutes. A multistage batch filtration process increased the lead removal to 50-54ppb/3g of foam over a period of 90 minutes.

Keywords: adsorption, functionalized, ion exchange, polyurethane, sulfonic

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28723 Conversion of Glycerol to 3-Hydroxypropanoic Acid by Genetically Engineered Bacillus subtilis

Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

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3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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28722 Antimicrobial Activity of Fatty Acid Salts against Microbes for Food Safety

Authors: Aya Tanaka, Mariko Era, Manami Masuda, Yui Okuno, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives— Fungi and bacteria are present in a wide range of natural environments. They are breed in the foods such as vegetables and fruit, causing corruption and deterioration of these foods in some cases. Furthermore, some species of fungi and bacteria are known to cause food intoxication or allergic reactions in some individuals. To prevent fungal and bacterial contamination, various fungicides and bactericidal have been developed that inhibit fungal and bacterial growth. Fungicides and bactericides must show high antifungal and antibacterial activity, sustainable activity, and a high degree of safety. Therefore, we focused on the fatty acid salt which is the main component of soap. We focused on especially C10K and C12K. This study aimed to find the effectiveness of the fatty acid salt as antimicrobial agents for food safety. Materials and Methods— Cladosporium cladosporioides NBRC 30314, Penicillium pinophilum NBRC 6345, Aspergillus oryzae (Akita Konno store), Rhizopus oryzae NBRC 4716, Fusarium oxysporum NBRC 31631, Escherichia coli NBRC 3972, Bacillus subtilis NBRC 3335, Staphylococcus aureus NBRC 12732, Pseudomonas aenuginosa NBRC 13275 and Serratia marcescens NBRC 102204 were chosen as tested fungi and bacteria. Hartmannella vermiformis NBRC 50599 and Acanthamoeba castellanii NBRC 30010 were chosen as tested amoeba. Nine fatty acid salts including potassium caprate (C10K) and laurate (C12K) at 350 mM and pH 10.5 were used as antifungal activity. The spore suspension of each fungus (3.0×10⁴ spores/mL) or the bacterial suspension (3.0×10⁵ or 3.0×10⁶ or 3.0×10⁷ CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar or nutrient agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 °C. Results— C10K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than A. oryzae. C12K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than P. pinophilum and A. oryzae. C10K and C12K did not show high anti-yeast activity. C10K was antibacterial activity of 6 or 7 log-unit incubated time for 10 min against bacteria other than B. subtilis. C12K was antibacterial activity of 5 to 7 log-unit incubated time for 10 min against bacteria other than S. marcescens. C12K was anti-amoeba activity of 4 log-unit incubated time for 10 min against H. vermiformis. These results suggest C10K and C12K have potential in the field of food safety.

Keywords: food safety, microbes, antimicrobial, fatty acid salts

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28721 Mitigating Acid Mine Drainage Pollution: A Case Study In the Witwatersrand Area of South Africa

Authors: Elkington Sibusiso Mnguni

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In South Africa, mining has been a key economic sector since the discovery of gold in 1886 in the Witwatersrand region, where the city of Johannesburg is located. However, some mines have since been decommissioned, and the continuous pumping of acid mine drainage (AMD) also stopped causing the AMD to rise towards the ground surface. This posed a serious environmental risk to the groundwater resources and river systems in the region. This paper documents the development and extent of the environmental damage as well as the measures implemented by the government to alleviate such damage. The study will add to the body of knowledge on the subject of AMD treatment to prevent environmental degradation. The method used to gather and collate relevant data and information was the desktop study. The key findings include the social and environmental impact of the AMD, which include the pollution of water sources for domestic use leading to skin and other health problems and the loss of biodiversity in some areas. It was also found that the technical intervention of constructing a plant to pump and treat the AMD using the high-density sludge technology was the most effective short-term solution available while a long-term solution was being explored. Some successes and challenges experienced during the implementation of the project are also highlighted. The study will be a useful record of the current status of the AMD treatment interventions in the region.

Keywords: acid mine drainage, groundwater resources, pollution, river systems, technical intervention, high density sludge

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28720 Supplementation of Jackfruit By-Product Concentrate in Combination with Two Types of Protein Sources for Growing Kids

Authors: Emely J. Escala, Lolito C. Bestil

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An experiment was conducted to assess the potential of jackfruit by-product concentrate (JBC) in combination with two types of protein sources, soybean meal (SBM) and liquid acid whey (LAW), given at two different ratios as supplement for growing kids fed a basal diet of 70:30 napier grass (Pennisetum purpureum) and kakawate (Gliricidia sepium) soilage ratio. The experiment was set-up in randomized complete block design (RCBD) with sex-age combination as basis for blocking, with the following dietary treatments: T1 = 0.50:0.50% BW, DM basis, JBC:SBM, T2 = 0.75:0.25% BW JBC:SBM, T3 = 0.50:0.50% BW, DM basis, JBC:LAW, and T4 = 0.75:0.25% BW JBC:LAW. Analysis of JBC showed high contents of crude fiber with medium levels of crude protein and nitrogen-free extract, appearing to be fitting for ruminants and a potential energy source. Results showed significantly higher voluntary dry matter intake (VDMI), cumulative weight gain (CWG), and average daily gain (ADG) of growing goats supplemented with JBC in combination with SBM than with LAW. The amount of JBC can range from 0.50% to 0.75% BW with SBM making up the difference, but a JBC:SBM ratio of 0.75:0.25% BW, DM basis, is best in promoting highest voluntary dry matter intake and is, therefore, highly recommended in the light of savings in feed cost. A long-term study on the effects of JBC supplementation on meat qualities of growing kids (aroma, marbling characteristics and taste) is also recommended.

Keywords: jackfruit by-product concentrate, liquid acid whey, soybean meal, grower kids

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28719 Oil Extraction from Microalgae Dunalliela sp. by Polar and Non-Polar Solvents

Authors: A. Zonouzi, M. Auli, M. Javanmard Dakheli, M. A. Hejazi

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Microalgae are tiny photosynthetic plants. Nowadays, microalgae are being used as nutrient-dense foods and sources of fine chemicals. They have significant amounts of lipid, carotenoids, vitamins, protein, minerals, chlorophyll, and pigments. Oil extraction from algae is a hotly debated topic currently because introducing an efficient method could decrease the process cost. This can determine the sustainability of algae-based foods. Scientific research works show that solvent extraction using chloroform/methanol (2:1) mixture is one of the efficient methods for oil extraction from algal cells, but both methanol and chloroform are toxic solvents, and therefore, the extracted oil will not be suitable for food application. In this paper, the effect of two food grade solvents (hexane and hexane/ isopropanol) on oil extraction yield from microalgae Dunaliella sp. was investigated and the results were compared with chloroform/methanol (2:1) extraction yield. It was observed that the oil extraction yield using hexane, hexane/isopropanol (3:2) and chloroform/methanol (2:1) mixture were 5.4, 13.93, and 17.5 (% w/w, dry basis), respectively. The fatty acid profile derived from GC illustrated that the palmitic (36.62%), oleic (18.62%), and stearic acids (19.08%) form the main portion of fatty acid composition of microalgae Dunalliela sp. oil. It was concluded that, the addition of isopropanol as polar solvent could increase the extraction yield significantly. Isopropanol solves cell wall phospholipids and enhances the release of intercellular lipids, which improves accessing of hexane to fatty acids.

Keywords: fatty acid profile‎, microalgae‎, oil extraction‎, polar solvent‎

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28718 Binding Studies of Complexes of Anticancer Drugs with DNA and Enzymes Involved in DNA Replication Using Molecular Docking and Cell Culture Techniques

Authors: Fouzia Perveen, Rumana Qureshi

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The presently studied twelve anticancer drugs are the cytotoxic agents which inhibit the replication of DNA and activity of enzymes involved in DNA replication namely topoisomerase-II, polymerase and helicase and have shown remarkable anticancer activity in clinical trials. In this study, we performed molecular docking studies of twelve antitumor drugs against DNA and DNA enzymes in the presence and absence of ascorbic acid (AA) and developed the quantitative structure-activity relationship (QSAR) model for anticancer activity screening. A number of electronic and steric descriptors were calculated using MOE software package. QSAR was established showing a correlation of binding strength with various physicochemical descriptors. Out of these twelve, eight cytotoxic drugs were tested on Non-Small Cell Lung Cancer cell lines (H-157 and H-1299) in the absence and presence of ascorbic acid and experimental IC50 values were calculated. From the docking studies, binding constants were calculated indicating the strength of drug-DNA and drug-enzyme complex formation and it was correlated to the IC50 values (both experimental and theoretical). These results can offer useful references for directing the molecular design of DNA enzyme inhibitor with improved anticancer activity.

Keywords: ascorbic acid, binding constant, cytotoxic agents, cell culture, DNA, DNA enzymes, molecular docking

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28717 Enhancement and Characterization of Titanium Surfaces with Sandblasting and Acid Etching for Dental Implants

Authors: Busra Balli, Tuncay Dikici, Mustafa Toparli

Abstract:

Titanium and its alloys have been used extensively over the past 25 years as biomedical materials in orthopedic and dental applications because of their good mechanical properties, corrosion resistance, and biocompatibility. It is known that the surface properties of titanium implants can enhance the cellular response and play an important role in Osseo integration. The rate and quality of Osseo integration in titanium implants are related to their surface properties. The purpose of this investigation was to evaluate the effect of sandblasting and acid etching on surface morphology, roughness, the wettability of titanium. The surface properties will be characterized by scanning electron microscopy and contact angle and roughness measurements. The results show that surface morphology, roughness, and wettability were changed and enhanced by these treatments.

Keywords: dental implant, etching, surface modifications, surface morphology, surface roughness

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28716 Effects of Lipoic Acid Supplementation on Activities of Cyclooxygenases and Levels of Prostaglandins E2 and F2 Alpha Metabolites in the Offspring of Rats with Streptozocin-Induced Diabetes

Authors: H. Y. Al-Matubsi, G. A. Oriquat, M. Abu-Samak, O. A. Al Hanbali, M. Salim

Abstract:

Background: Uncontrolled diabetes mellitus (DM) is an etiological factor for recurrent pregnancy loss and major congenital malformations in the offspring. Antioxidant therapy has been advocated to overcome the oxidant-antioxidant disequilibrium inherent in diabetes. The aims of this study were to evaluate the protective effect of lipoic acid (LA) on fetal outcome and to elucidate changes that may be involved in the mechanism(s) implicit diabetic fetopathy. Methods: Female rats were rendered hyperglycemic using streptozocin and then mated with normal male rat. Pregnant non-diabetic (group1; n=9; and group2; n=7) or pregnant diabetic (group 3; n=10; and group 4; n=8) rats were treated daily with either lipoic acid (LA) (30 mg/kg body weight; groups 2 and 4) or vehicle (groups 1 and 3) between gestational days 0 and 15. On day 15 of gestation, the rats were sacrificed, and the fetuses, placentas and membranes dissected out of the uterine horns. Following morphological examination, the fetuses, placentas and membranes were homogenized, and used to measure cyclooxygenases (COX) activities and metabolisms of prostaglandin (PG) E2 (PGEM) and PGF2 (PGFM) levels. Maternal liver and plasma total glutathione levels were also determined. Results: Supplementation of diabetic rats with LA was found to significantly (P<0.05) reduce resorption rates in diabetic rats and increased mean fetal weight compared to diabetic group. Treatment of diabetic rats with LA leads to a significant (P<0.05) increase in liver and plasma total glutathione, in comparison with diabetic rats. Decreased levels of PGEM and elevated levels of PGFM in the fetuses, placentas and membranes were characteristic of experimental diabetic gestation associated with malformation. LA treatment to diabetic mothers failed to normalize levels of PGEM to the non-diabetic control rats. However, the levels of PGEM in malformed fetuses from LA-treated diabetic mothers was significantly (P < 0.05) higher than those in malformed fetuses from diabetic rats. Conclusions: We conclude that LA can reduce congenital malformations in the offspring of diabetic rats at day 15 of gestation. However, LA treatment did not completely prevent the occurrence of malformations, other factors, such as arachidonic acid deficiency and altered prostaglandin metabolismmay be involved in the pathogenesis of diabetes-induced congenital malformations.

Keywords: diabetes, lipoic acid, pregnancy, prostaglandins

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28715 Green Synthesis of Red-Fluorescent Gold Nanoclusters: Characterization and Application for Breast Cancer Detection

Authors: Agnė Mikalauskaitė, Renata Karpicz, Vitalijus Karabanovas, Arūnas Jagminas

Abstract:

The use of biocompatible precursors for the synthesis and stabilization of fluorescent gold nanoclusters (NCs) with strong red photoluminescence creates an important link between natural sciences and nanotechnology. Herein, we report the cost-effective synthesis of Au nanoclusters by templating and reduction of chloroauric acid with the cheap amino acid food supplements. This synthesis under the optimized conditions leads to the formation of biocompatible Au NCs having good stability and intense red photoluminescence, peaked at 680 to 705 nm, with a quantum yield (QY) of ≈7% and the average lifetime of up to several µs. The composition and luminescent properties of the obtained NCs were compared with ones formed via well-known bovine serum albumin reduction approach. Our findings implied that synthesized Au NCs tend to accumulate in more tumorigenic breast cancer cells (line MDA-MB-213) and after dialysis can be prospective for bio imagining.

Keywords: gold nanoclusters, proteins, materials chemistry, red-photoluminescence, bioimaging

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28714 Carbon Nanomaterials from Agricultural Wastes for Adsorption of Organic Pollutions

Authors: Magdalena Blachnio, Viktor Bogatyrov, Mariia Galaburda, Anna Derylo-Marczewska

Abstract:

Agricultural waste materials from traditional oil mill and after extraction of natural raw materials in supercritical conditions were used for the preparation of carbon nanomaterials (activated carbons) by two various methods. Chemical activation using acetic acid and physical activation with a gaseous agent (carbon dioxide) were chosen as mild and environmentally friendly ones. The effect of influential factors: type of raw material, temperature and activation agent on the porous structure characteristics of the materials was discussed by using N₂ adsorption/desorption isotherms at 77 K. Furthermore scanning electron microscope (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) were employed to examine the physicochemical properties of the obtained sorbents. Selection of a raw material and an optimization of the conditions of the synthesis process, allowed to obtain the cheap sorbents with a targeted distribution of pores enabling effective adsorption of the model organic pollutants carried out in the multicomponent systems. Adsorption behavior (capacity and rate) of the chosen activated carbons was estimated by utilizing Crystal violet (CV), 4-chlorophenoxyacetic acid (4-CPA), 2.4-dichlorophenoxyacetic acid (2.4-D) as the adsorbates. Both rate and adsorption capacity of the organics on the sorbents evidenced that the activated carbons could be effectively used in sewage treatment plants. The mechanisms of organics adsorption were studied and correlated with activated carbons properties.

Keywords: activated carbon, adsorption equilibrium, adsorption kinetics, organics adsorption

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28713 Elaboration of Composites with Thermoplastic Matrix Polypropylene Charged by the Polyaniline Synthesized by the Self-Curling Method

Authors: Selma Saadia, Nacira Naar, Ahmed Benaboura

Abstract:

This work is dedicated to the elaboration of composites (PP/PANI) with Polypropylene (PP) as thermoplastic polymer and the polyaniline (PANI) as electric charge doped with sulfanilic acid (PANI-As). These realized formulations are intended for the antistatic domain. The used conductive polymer is synthesized by the method self-curling which proved the obtaining of the nanoparticles of PANI in regular morphological forms. The PANI and PP composites are fabricated into a film by a twin-screw extruding. Several methods of characterization are proposed: spectroscopic, thermal, and electric. The realized composites proved a pseudo-homogeneous aspect and the threshold percolation study, showed that the formulation with 7% of PANI presents a better formulation which can be used in the antistatic domain.

Keywords: extruding, PANI, Polypropylene, sulfanilic acid, self-Curling

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28712 Bioinformatics Approach to Identify Physicochemical and Structural Properties Associated with Successful Cell-free Protein Synthesis

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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28711 Anti-Obesity Effects of Pteryxin in Peucedanum japonicum Thunb Leaves through Different Pathways of Adipogenesis In-Vitro

Authors: Ruwani N. Nugara, Masashi Inafuku, Kensaku Takara, Hironori Iwasaki, Hirosuke Oku

Abstract:

Pteryxin from the partially purified hexane phase (HP) of Peucedanum japonicum Thunb (PJT) was identified as the active compound related to anti-obesity. Thus, in this study we investigated the mechanisms related to anti-obesity activity in-vitro. The HP was fractionated, and effect on the triglyceride (TG) content was evaluated in 3T3-L1 and HepG2 cells. Comprehensive spectroscopic analyses were used to identify the structure of the active compound. The dose dependent effect of active constituent on the TG content, and the gene expressions related to adipogenesis, fatty acid catabolism, energy expenditure, lipolysis and lipogenesis (20 μg/mL) were examined in-vitro. Furthermore, higher dosage of pteryxin (50μg/mL) was tested against 20μg/mL in 3T3-L1 adipocytes. The mRNA were subjected to SOLiD next generation sequencer and the obtained data were analyzed by Ingenuity Pathway Analysis (IPA). The active constituent was identified as pteryxin, a known compound in PJT. However, its biological activities against obesity have not been reported previously. Pteryxin dose dependently suppressed TG content in both 3T3-L1 adipocytes and HepG2 hepatocytes (P < 0.05). Sterol regulatory element-binding protein-1 (SREBP1 c), Fatty acid synthase (FASN), and acetyl-CoA carboxylase-1 (ACC1) were downregulated in pteryxin-treated adipocytes (by 18.0, 36.1 and 38.2%; P < 0.05, respectively) and hepatocytes (by 72.3, 62.9 and 38.8%, respectively; P < 0.05) indicating its suppressive effects on fatty acid synthesis. The hormone-sensitive lipase (HSL), a lipid catabolising gene was upregulated (by 15.1%; P < 0.05) in pteryxin-treated adipocytes suggesting improved lipolysis. Concordantly, the adipocyte size marker gene, paternally expressed gene1/mesoderm specific transcript (MEST) was downregulated (by 42.8%; P < 0.05), further accelerating the lipolytic activity. The upregulated trend of uncoupling protein 2 (UCP2; by 77.5%; P < 0.05) reflected the improved energy expenditure due to pteryxin. The 50μg/mL dosage of pteryxin completely suppressed PPARγ, MEST, SREBP 1C, HSL, Adiponectin, Fatty Acid Binding Protein (FABP) 4, and UCP’s in 3T3-L1 adipocytes. The IPA suggested that pteryxin at 20μg/mL and 50μg/mL suppress obesity in two different pathways, whereas the WNT signaling pathway play a key role in the higher dose of pteryxin in preadipocyte stage. Pteryxin in PJT play the key role in regulating lipid metabolism related gene network and improving energy production in vitro. Thus, the results suggests pteryxin as a new natural compound to be used as an anti-obesity drug in pharmaceutical industry.

Keywords: obesity, peucedanum japonicum thunb, pteryxin, food science

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28710 Evaluation of Barium Sulfate and Its Surface Modification as Reinforcing Filler for Natural and Some Synthetic Rubbers

Authors: Mohamad Abdelfattah Ibrahim Elghrbawy

Abstract:

This work deals to evaluate barium sulfate (BS) before and after its surface modification as reinforcing filler for rubber. Barium sulfate was surface-modified using polymethacrylic acid (PMAA), the monolayer surface coverage of barium sulfate by polymethacrylic acid molecules occurred at 5.4x10-6 mol/g adsorbed amount. This amount was sufficient to reduce the sediment volume from 2.65 to 2.55 cm3/gm. Natural rubber (NR) was compounded with different concentrations of barium sulfate. The rheological characteristics of NR mixes were measured using a Monsanto Oscillating Disk Rheometer. The compounded NR was vulcanized at 142°C, and the physico-mechanical properties were tested according to the standard methods. The rheological data show that the minimum torque decreases while the maximum torque increases as the barium sulfate content increase. The physico-mechanical properties of NR vulcanizates were improved up to 50 phr/ barium sulfate loading. On the other hand, styrene–butadiene rubber (SBR) and nitrile–butadiene rubber (NBR) rubbers compounded with 50 phr/barium sulfate had good rheological and mechanical properties. Scanning electron microscope studies show surface homogeneity of rubber samples as a result of good dispersion of surface modified barium sulfate in the rubber matrix. The NR, SBR and NBR vulcanizates keep their values of mechanical properties after subjected to thermal oxidative aging at 90°C for 7 days.

Keywords: barium sulfate, natural rubber (nr), nitrile–butadiene rubber (nbr), polymethacrylic acid (pmaa), styrene–butadiene rubber (sbr), surface modification

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28709 Optical Characterization of Lead Sulphide Thin Films Grown by Chemical Bath Deposition

Authors: Ekpekpo Arthur

Abstract:

Thin films can either be conductive or dielectric (non-conductive). It is formed through atom/molecules state or formed after decomposing the materials into atomic/molecular scale by physical or chemical processes. In this study, thin films of Lead Sulphide were deposited on glass substrate prepared from lead acetate and thiourea solution using chemical bath deposition (CBD). The glass slides were subjected to the pretreatment by soaking them in a solution of 50% sulphuric acid and 50% nitric acid. Lead sulphide was deposited at different parameters such as deposition time and temperature. The optical properties of the thin films were determined from spectroscopy measurements of absorbance and reflectance. Optical studies show that the band gap of lead sulphide ranges between 0.41 eV to 300K.

Keywords: lead sulphide, spectroscopy, absorbance, reflectance

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28708 Structure-Based Virtual Screening and in Silico Toxicity Test of Compounds against Mycobacterium tuberculosis 7,8-Diaminopelargonic Acid Aminotransferase (MtbBioA)

Authors: Junie B. Billones, Maria Constancia O. Carrillo, Voltaire G. Organo, Stephani Joy Y. Macalino, Inno A. Emnacen, Jamie Bernadette A. Sy

Abstract:

One of the major interferences in the Philippines’ tuberculosis control program is the widespread prevalence of Mtb strains that are resistant to known drugs, such as the MDR-TB (Multi Drug Resistant Tuberculosis) and XDR-TB (Extensively Drug Resistant Tuberculosis). Therefore, there is a pressing need to search for novel Mtb drug targets in order to be able to combat these drug resistant strains. The enzyme 7,8-diaminopelargonic acid aminotransferase enzyme, or more commonly known as BioA, is one such ideal target, as it is known that humans do not possess this enzyme. BioA primarily plays a key role in Mtb’s lipid biosynthesis pathway; more specifically in the synthesis of the enzyme cofactor biotin. In this study, structure-based pharmacophore screening, docking, and ADMET evaluation of compounds obtained from the DrugBank chemical database were performed against the MtbBioA enzyme. Results of the screening, docking, ADMET, and TOPKAT calculations revealed that out of the 6,516 compounds in the library, only 7 compounds indicated more favorable binding energies as compared to the enzyme’s known inhibitor, amiclenomycin (ACM), as well as good solubility and toxicity properties. Moreover, out of these 7 compounds, Molecule 6 exhibited the best solubility and toxicity properties. In the future, these lead compounds may then be subjected to bioactivity assays in vitro or in vivo for further evaluation of its therapeutic efficacy.

Keywords: 7, 8-diaminopelargonic acid aminotransferase, BioA, pharmacophore, molecular docking, ADMET, TOPKAT

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28707 Fertilizer Value of Nitrogen Captured from Poultry Facilities Using Ammonia Scrubbers

Authors: Philip A. Moore Jr., Jerry Martin, Hong Li

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Research has shown that over half of the nitrogen (N) excreted from broiler chickens is emitted to the atmosphere before the manure is removed from the barns, resulting in air and water pollution, as well as the loss of a valuable fertilizer resource. The objective of this study was to determine the fertilizer efficiency of N captured from the exhaust air from poultry houses using acid scrubbers. This research was conducted using 24 plots located on a Captina silt loam soil. There were six treatments: (1) unfertilized control, (2) aluminum sulfate (alum) scrubber solution, (3) potassium bisulfate scrubber solution, (4) sodium bisulfate scrubber solution, (5) sulfuric acid scrubber solution and (6) ammonium nitrate fertilizer dissolved in water. There were four replications per treatment in a randomized block design. The scrubber solutions were obtained from acid scrubbers attached to exhaust fans on commercial broiler houses. All N sources were applied at an application rate equivalent to 112 kg N ha⁻¹. Forage yields were measured five times throughout the growing season. Five months after the fertilizer sources were applied, a rainfall simulation study was conducted to determine the potential effects on phosphorus (P) runoff. Forage yields were significantly higher in plots fertilized with scrubber solutions from potassium bisulfate and sodium bisulfate than plots fertilized with scrubber solutions made from alum or sulfuric acid or ammonium nitrate, which were higher than the controls (7.61, 7.46, 6.87, 6.72, 6.45, and 5.12 Mg ha ⁻¹, respectively). Forage N uptake followed similar trends as yields. Phosphorus runoff and water soluble P was significantly lower in plots fertilized with the scrubber solutions made from aluminum sulfate. This study demonstrates that N captured using ammonia scrubbers is as good or possibly better than commercial ammonium nitrate fertilizer.

Keywords: air quality, ammonia emissions, nitrogen fertilizer, poultry

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28706 Genetic Polymorphism and Insilico Study Epitope Block 2 MSP1 Gene of Plasmodium falciparum Isolate Endemic Jayapura

Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

Abstract:

Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

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