Search results for: ultrasound assisted acid hydrolysis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4562

Search results for: ultrasound assisted acid hydrolysis

1022 Perfluoroheptanoic Acid Affects Xenopus Embryo Embryogenesis by Inducing the Phosphorylation of ERK and JNK

Authors: Chowon Kim, Yoo-Kyung Kim, Kyeong Yeon Park, Hyun-Shik Lee

Abstract:

Perfluoroalkyl compounds (PFCs) are globally distributed synthetic compounds that are known to adversely affect human health. Developmental toxicity assessment of PFCs is important to facilitate the evaluation of their environmental impact. In the present study, we assessed the developmental toxicity and teratogenicity of PFCs with different numbers of carbon atoms on Xenopus embryogenesis. An initial frog embryo teratogenicity assay-Xenopus (FETAX) assay was performed that identified perfluorohexanoic (PFHxA) and perfluoroheptanoic (PFHpA) acids as potential teratogens and developmental toxicants. The mechanism underlying this teratogenicity was also investigated by measuring the expression of tissue-specific biomarkers such as phosphotyrosine‑binding protein, xPTB (liver); NKX2.5 (heart); and Cyl18 (intestine). Whole‑mount in situ hybridization, reverse transcriptase‑polymerase chain reaction (RT-PCR), and histologic analyses detected severe defects in the liver and heart following exposure to PFHxA or PFHpA. In addition, immunoblotting revealed that PFHpA significantly increased the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), while PFHxA slightly increased these, as compared with the control. These results suggest that PFHxA and PFHpA are developmental toxicants and teratogens, with PFHpA producing more severe effects on liver and heart development through the induction of ERK and JNK phosphorylation.

Keywords: PFCs, ERK, JNK, xenopus

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1021 RP-HPLC Method Development and Its Validation for Simultaneous Estimation of Metoprolol Succinate and Olmesartan Medoxomil Combination in Bulk and Tablet Dosage Form

Authors: S. Jain, R. Savalia, V. Saini

Abstract:

A simple, accurate, precise, sensitive and specific RP-HPLC method was developed and validated for simultaneous estimation of Metoprolol Succinate and Olmesartan Medoxomil in bulk and tablet dosage form. The RP-HPLC method has shown adequate separation for Metoprolol Succinate and Olmesartan Medoxomil from its degradation products. The separation was achieved on a Phenomenex luna ODS C18 (250mm X 4.6mm i.d., 5μm particle size) with an isocratic mixture of acetonitrile: 50mM phosphate buffer pH 4.0 adjusted with glacial acetic acid in the ratio of 55:45 v/v. The mobile phase at a flow rate of 1.0ml/min, Injection volume 20μl and wavelength of detection was kept at 225nm. The retention time for Metoprolol Succinate and Olmesartan Medoxomil was 2.451±0.1min and 6.167±0.1min, respectively. The linearity of the proposed method was investigated in the range of 5-50μg/ml and 2-20μg/ml for Metoprolol Succinate and Olmesartan Medoxomil, respectively. Correlation coefficient was 0.999 and 0.9996 for Metoprolol Succinate and Olmesartan Medoxomil, respectively. The limit of detection was 0.2847μg/ml and 0.1251μg/ml for Metoprolol Succinate and Olmesartan Medoxomil, respectively and the limit of quantification was 0.8630μg/ml and 0.3793μg/ml for Metoprolol and Olmesartan, respectively. Proposed methods were validated as per ICH guidelines for linearity, accuracy, precision, specificity and robustness for estimation of Metoprolol Succinate and Olmesartan Medoxomil in commercially available tablet dosage form and results were found to be satisfactory. Thus the developed and validated stability indicating method can be used successfully for marketed formulations.

Keywords: metoprolol succinate, olmesartan medoxomil, RP-HPLC method, validation, ICH

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1020 Synthesis and Characterization of Un-Doped and Velvet Tamarind Doped ZnS Crystals, Using Sol Gel Method

Authors: Uchechukwu Vincent Okpala

Abstract:

Under the Sun, energy is a key factor for the sustenance of life and its environment. The need to protect the environment as energy is generated and consumed has called for renewable and green energy sources. To be part of this green revolution, we synthesized and characterized undoped and velvet tamarind doped zinc sulfide (ZnS) crystals using sol-gel methods. Velvet tamarind was whittled down using the top-down approach of nanotechnology. Sodium silicate, tartaric acid, zinc nitrate, and thiourea were used as precursors. The grown samples were annealed at 105°C. Structural, optical, and compositional analyses of the grown samples revealed crystalline structures with varied crystallite sizes influenced by doping. Energy-dispersive X-ray spectroscopy confirmed elemental compositions of Zn, S, C and O in the films. Atomic percentages of the elements varied with VT doping. FT-IR analysis indicated the presence of functional groups like O-H stretching (alcohol), C=C=C stretching (alkene group), C=C bending, C-H stretching (alkane), N-H stretching (aliphatic primary amine) and N=C=S stretching (isothiocyanate) constituent in the film. The transmittance of the samples increased from the visible region to the infrared region making the samples good for poultry and solar energy applications. The bandgap energy of the films decreased as the number of VT drops increased, from 2.4 to 2.2. They were wide band gap materials and were good for optoelectronic, photo-thermal, high temperature, high power and solar cell applications.

Keywords: doping, sol-gel, velvet tamarind, ZnS.

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1019 Nanosilver Containing Biodegradable Bionanocomposites for Antimicrobial Application: Design, Preparation and Study

Authors: Nino Kupatadze, Shorena Tskhadadze, Mzevinar Bedinashvili, David Tugushi, Ramaz Katsarava

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Surgical device-associated infection and biofilm formation are some of the major problems in biomedicine for today. The losing protection ability of conventional antimicrobial-drugs leads to the challenges in the current antibiotic therapy, the most serious of which is antibiotic resistance. Our strategy to overcome the biofilm formation consists in coating devices with polymeric film containing nanosilver(AgNPs) as a bactericidal agent. Such bionanocomposites are also promising as wound dressing materials. For this purpose, we have developed a new generation of AgNPs containing polymeric composites in which amino acid based biodegradable poly(ester amide)s (PEAs) were served as both matrices and AgNPs stabilizers. The AgNPs were formed by photochemical (daylight) reduction of AgNO3 in ethanol solution. The formation of AgNPs was monitored by coloring the solution in brownish-red and appearance of the absorption maximum at 420-430 nm in UV spectrum. Comparative studies of PEAs with polyvinylpyrrolidone (PVP) as particle stabilizers were carried out. It was found that PVP is better stabilizer in terms of particles yield and stability. Therefore, in subsequent experiments blends of PEAs and PVP were used as stabilizers for fabricating AgNPs. As expected, PVP increased the stabilizing effect and this apparently observed in the UV spectrum of the samples after 7 h daylight irradiation: for pure PVP λmax = 430 nm, D = 2.03, for pure PEA λmax= 420 nm, D = 0.65, and for the blend of PVP and PEA λmax = 435 nm, D = 1.88. Further study of the obtained nanobiocomposites is in progress now.

Keywords: biodegradation, bionanocompositions, polymer, nanosilver

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1018 Electrochemical Corrosion and Mechanical Properties of Structural Materials for Oil and Gas Applications in Simulated Deep-Sea Well Environments

Authors: Turin Datta, Kisor K. Sahu

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Structural materials used in today’s oil and gas exploration and drilling of both onshore and offshore oil and gas wells must possess superior tensile properties, excellent resistance to corrosive degradation that includes general, localized (pitting and crevice) and environment assisted cracking such as stress corrosion cracking and hydrogen embrittlement. The High Pressure and High Temperature (HPHT) wells are typically operated at temperature and pressure that can exceed 300-3500F and 10,000psi (69MPa) respectively which necessitates the use of exotic materials in these exotic sources of natural resources. This research investigation is focussed on the evaluation of tensile properties and corrosion behavior of AISI 4140 High-Strength Low Alloy Steel (HSLA) possessing tempered martensitic microstructure and Duplex 2205 Stainless Steel (DSS) having austenitic and ferritic phase. The selection of this two alloys are primarily based on economic considerations as 4140 HSLA is cheaper when compared to DSS 2205. Due to the harsh aggressive chemical species encountered in deep oil and gas wells like chloride ions (Cl-), carbon dioxide (CO2), hydrogen sulphide (H2S) along with other mineral organic acids, DSS 2205, having a dual-phase microstructure can mitigate the degradation resulting from the presence of both chloride ions (Cl-) and hydrogen simultaneously. Tensile properties evaluation indicates a ductile failure of DSS 2205 whereas 4140 HSLA exhibit quasi-cleavage fracture due to the phenomenon of ‘tempered martensitic embrittlement’. From the potentiodynamic polarization testing, it is observed that DSS 2205 has higher corrosion resistance than 4140 HSLA; the former exhibits passivity signifying resistance to localized corrosion while the latter exhibits active dissolution in all the environmental parameters space that was tested. From the Scanning Electron Microscopy (SEM) evaluation, it is understood that stable pits appear in DSS 2205 only when the temperature exceeds the critical pitting temperature (CPT). SEM observation of the corroded 4140 HSLA specimen tested in aqueous 3.5 wt.% NaCl solution reveals intergranular cracking which appears due to the adsorption and diffusion of hydrogen during polarization, thus, causing hydrogen-induced cracking/hydrogen embrittlement. General corrosion testing of DSS 2205 in acidic brine (pH~3.0) solution at ambient temperature using coupons indicate no weight loss even after three months whereas the corrosion rate of AISI 4140 HSLA is significantly higher after one month of testing.

Keywords: DSS 2205, polarization, pitting, SEM

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1017 Epigenetic Drugs for Major Depressive Disorder: A Critical Appraisal of Available Studies

Authors: Aniket Kumar, Jacob Peedicayil

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Major depressive disorder (MDD) is a common and important psychiatric disorder. Several clinical features of MDD suggest an epigenetic basis for its pathogenesis. Since epigenetics (heritable changes in gene expression not involving changes in DNA sequence) may underlie the pathogenesis of MDD, epigenetic drugs such as DNA methyltransferase inhibitors (DNMTi) and histone deactylase inhibitors (HDACi) may be useful for treating MDD. The available literature indexed in Pubmed on preclinical drug trials of epigenetic drugs for the treatment of MDD was investigated. The search terms we used were ‘depression’ or ‘depressive’ and ‘HDACi’ or ‘DNMTi’. Among epigenetic drugs, it was found that there were 3 preclinical trials using HDACi and 3 using DNMTi for the treatment of MDD. All the trials were conducted on rodents (mice or rats). The animal models of depression that were used were: learned helplessness-induced animal model, forced swim test, open field test, and the tail suspension test. One study used a genetic rat model of depression (the Flinders Sensitive Line). The HDACi that were tested were: sodium butyrate, compound 60 (Cpd-60), and valproic acid. The DNMTi that were tested were: 5-azacytidine and decitabine. Among the three preclinical trials using HDACi, all showed an antidepressant effect in animal models of depression. Among the 3 preclinical trials using DNMTi also, all showed an antidepressant effect in animal models of depression. Thus, epigenetic drugs, namely, HDACi and DNMTi, may prove to be useful in the treatment of MDD and merit further investigation for the treatment of this disorder.

Keywords: DNA methylation, drug discovery, epigenetics, major depressive disorder

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1016 Characterization of Solanum tuberosum Ammonium Transporter Gene Using Bioinformatics Approach

Authors: Adewole Tomiwa Adetunji, Francis Bayo Lewu, Richard Mundembe

Abstract:

Plants require nitrogen (N) to support desired production levels. There is a need for better understanding of N transport mechanism in order to improve N assimilation by plant root. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which N is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was amplified, sequenced and characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design 976 base pairs AMT1-specific primers which include forward primer 5’- GCCATCGCCGCCGCCGG-3’ and reverse primer 5’-GGGTCAGATCCATACCCGC-3’. These primers were used to amplify the Solanum tuberosum AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family due to the clade and high similarity it shared with other plant AMT1 genes. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1, and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th-10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.

Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum

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1015 Molecular Characterization of Functional Domain (LRR) of TLR9 Genes in Malnad Gidda Cattle and Their Comparison to Cross Breed Cattle

Authors: Ananthakrishna L. R., Ramesh D., Kumar Wodeyar, Kotresh A. M., Gururaj P. M.

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Malnad Gidda is the indigenous recognized cattle breed of Shivamogga District of Karnataka state, India is known for its disease resistance to many of the infectious diseases. There are 25 LRR (Leucine Rich Repeats) identified in bovine (Bos indicus) TLR9. The amino acid sequence of LRR is deduced to nucleotide sequence in BLASTx bioinformatic online tools. LRR2 to LRR10 are involved in pathogen recognition and binding in human TLR9 which showed a higher degree of nucleotide variations with respect to disease resistance to various pathogens. Hence, primers were designed to amplify the flanking sequences of LRR2 to LRR10, to discover the nucleotide variations if any, in Malnad Gidda breed of Cattle which is associated with disease resistance. The DNA isolated from peripheral blood mononuclear cells of ten Malnad Gidda cattle. A desired and specific amplification product of 0.8 kb was obtained at an annealing temperature of 56.6ᵒC. All the PCR products were sequenced on both sides by gene-specific primers. The sequences were compared with TLR9 sequence of cross breed cattle obtained from NCBI data bank. The sequence analysis between Malnad Gidda and crossbreed cattle revealed no nucleotide variations in the region LRR2 to LRR9 which shows the conserved in pathogen binding domain (LRR) of TLR9.

Keywords: leucine rich repeats, Malnad Gidda, cross breed, TLR9

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1014 Authenticity of Lipid and Soluble Sugar Profiles of Various Oat Cultivars (Avena sativa)

Authors: Marijana M. Ačanski, Kristian A. Pastor, Djura N. Vujić

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The identification of lipid and soluble sugar components in flour samples of different cultivars belonging to common oat species (Avena sativa L.) was performed: spring oat, winter oat and hulless oat. Fatty acids were extracted from flour samples with n-hexane, and derivatized into volatile methyl esters, using TMSH (trimethylsulfonium hydroxide in methanol). Soluble sugars were then extracted from defatted and dried samples of oat flour with 96% ethanol, and further derivatized into corresponding TMS-oximes, using hydroxylamine hydrochloride solution and BSTFA (N,O-bis-(trimethylsilyl)-trifluoroacetamide). The hexane and ethanol extracts of each oat cultivar were analyzed using GC-MS system. Lipid and simple sugar compositions are very similar in all samples of investigated cultivars. Chemometric tool was applied to numeric values of automatically integrated surface areas of detected lipid and simple sugar components in their corresponding derivatized forms. Hierarchical cluster analysis shows a very high similarity between the investigated flour samples of oat cultivars, according to the fatty acid content (0.9955). Moderate similarity was observed according to the content of soluble sugars (0.50). These preliminary results support the idea of establishing methods for oat flour authentication, and provide the means for distinguishing oat flour samples, regardless of the variety, from flour samples made of other cereal species, just by lipid and simple sugar profile analysis.

Keywords: oat cultivars, lipid composition, soluble sugar composition, GC-MS, chemometrics, authentication

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1013 Application of Synthetic Monomers Grafted Xanthan Gum for Rhodamine B Removal in Aqueous Solution

Authors: T. Moremedi, L. Katata-Seru, S. Sardar, A. Bandyopadhyay, E. Makhado, M. Joseph Hato

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The rapid industrialisation and population growth have led to a steady fall in freshwater supplies worldwide. As a result, water systems are affected by modern methods upon use due to secondary contamination. The application of novel adsorbents derived from natural polymer holds a great promise in addressing challenges in water treatment. In this study, the UV irradiation technique was used to prepare acrylamide (AAm) monomer, and acrylic acid (AA) monomer grafted xanthan gum (XG) copolymer. Furthermore, the factors affecting rhodamine B (RhB) adsorption from aqueous media, such as pH, dosage, concentration, and time were also investigated. The FTIR results confirmed the formation of graft copolymer by the strong vibrational bands at 1709 cm-1 and 1612 cm-1 for AA and AAm, respectively. Additionally, more irregular, porous and wrinkled surface observed from SEM of XG-g-AAm/AA indicated copolymerization interaction of monomers. The optimum conditions for removing RhB dye with a maximum adsorption capacity of 313 mg/g at 25 0C from aqueous solution were pH approximately 5, initial dye concentration = 200 ppm, adsorbent dose = 30 mg. Also, the detailed investigation of the isothermal and adsorption kinetics of RhB from aqueous solution showed that the adsorption of the dye followed a Freundlich model (R2 = 0.96333) and pseudo-second-order kinetics. The results further indicated that this absorbent based on XG had the universality to remove dye through the mechanism of chemical adsorption. The outstanding adsorption potential of the grafted copolymer could be used to remove cationic dyes from aqueous solution as a low-cost product.

Keywords: xanthan gum, adsorbents, rhodamine B, Freundlich

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1012 Microstructure and Tribological Properties of AlSi5Cu2/SiC Composite

Authors: Magdalena Suśniak, Joanna Karwan-Baczewska

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Microstructure and tribological properties of AlSi5Cu2 matrix composite reinforced with SiC have been studied by microscopic examination and basic tribological properties. Composite material was produced by the mechanical alloying and spark plasma sintering (SPS) technique. The mixture of AlSi5Cu2 chips with 0, 10, 15 wt. % of SiC powder were placed in 250 ml mixing jar and milled 40 hours. To prevent the extreme cold welding the 1 wt. % of stearic acid was added to the powder mixture as a process control agent. Mechanical alloying provide to obtain composites powder with uniform distribution of SiC in matrix. Composite powders were poured into a graphite and a pulsed electric current was passed through powder under vacuum to consolidate material. Processing conditions were: sintering temperature 450°C, uniaxial pressure 32MPa, time of sintering 5 minutes. After SPS process composite samples indicate higher hardness values, lower weight loss, and lower coefficient of friction as compared with the unreinforced alloy. Light microscope micrograph of the worn surfaces and wear debris revealed that in the unreinforced alloy the prominent wear mechanism was the adhesive wear. In the AlSi5Cu2/SiC composites, by increasing of SiC the wear mechanism changed from adhesive and micro-cutting to abrasive and delamination for composite with 20 SiC wt. %. In all the AlSi5Cu2/SiC composites, abrasive wear was the main wear mechanism.

Keywords: aluminum matrix composite, mechanical alloying, spark plasma sintering, AlSi5Cu2/SiC composite

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1011 The Effect of Different Concentrations of Trichoderma harzianum Fungus on the Phytochemical and Antioxidative Parameters of Cauliflower (Brassica oleracea convar.botrytisl) in Soils Contaminated with Lead

Authors: Mohammad Javad Shakori, Esmaeil Babakhanzadeh Sajirani, Vajihe Esmaili

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Today, the increasing contamination is an environmental concern. There is relationship between plants and microorganisms many years ago. In this regard, an experiment was conducted in order to investigate the effect of different levels of lead across three levels ‘zero, 50, and 100 mg/L’ and Trichoderma Harzanium fungus across three levels ‘5, 10, and 15%’ in a factorial design in the form of fully randomized blocks in three replications under form conditions in the climatic conditions of Shahroud in Dehlama Village. This research was performed in 2014-2015 on cauliflower. In this experiment, chlorophyll a, b, total, cartenoid, phenol, flavonoid, and antioxidant properties of cauliflowers were measured. The results indicated that the greatest level of chlorophyll a (75.723 mg/wet weight), chlorophyll b (27.378 mg/wet weight), and total chlorophyll (109.074 mg/wet weight) was related to the interactive effects of 5% treatment of Trichoderma fungus and 0mg/L lead. The results also indicated that the greatest amount of antioxidant (79.88% of free radical) and flavonoides (22.889 mg of coercetin/g of dry weight) was related to the interactive effects of lead 50 mg/L and the treatment of Trichoderma fungus 5%. Further, the greatest level of phenol (21.33 mg of Gaelic acid/ dry weight) was related to the interactive effects of lead 100 mg/L and Trichoderma fungus 5% . As carotenoids are a type of antioxidant and precursor of vitamin A, with the development of alignment effect with other antioxidants such as the total phenol, flavonoid, achieved desirable levels of antioxidant.

Keywords: antioxidant, lead, flavonoid, cauliflower, chlorophyll

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1010 Application of Active Chitosan Coating Incorporated with Spirulina Extract as a Potential Food Packaging Material for Enhancing Quality and Shelf Life of Shrimp

Authors: Rafik Balti, Nourhene Zayoud, Mohamed Ben Mansour, Abdellah Arhaliass, Anthony Masse

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Application of edible films and coatings with natural active compounds for enhancing storage stability of food products is a promising active packaging approach. Shrimp are generally known as valuable seafood products around the world because of their delicacy and good nutritional. However, shrimp is highly vulnerable to quality deterioration associated with biochemical, microbiological or physical changes during postmortem storage, which results in the limited shelf life of the product. Chitosan is considered as a functional packaging component for maintaining the quality and increasing the shelf life of perishable foods. The present study was conducted to evaluate edible coating of crab chitosan containing variable levels of ethanolic extract of Spirulina on microbiological (mesophilic aerobic, psychrotrophic, lactic acid bacteria, and enterobacteriacea), chemical (pH, TVB-N, TMA-N, PV, TBARS) and sensory (odor, color, texture, taste, and overall acceptance) properties of shrimp during refrigerated storage. Also, textural and color characteristics of coated shrimp were performed. According to the obtained results, crab chitosan in combination with Spirulina extract was very effective in order to extend the shelf life of shrimp during storage in refrigerated condition.

Keywords: food packaging, chitosan, spirulina extract, white shrimp, shelf life

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1009 Antioxidant and Antimicrobial Activities of Phenolic Extracts of Endemic Plants Marrubium deserti and Ammodaucus leucotrichus from Algeria

Authors: Sifi Ibrahim, Benaddou Fatima Zohra, Yousfi Mohamed

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The Marrubium deserti and Ammodaucus leucotrichus L. an Algerian endemic species, has several applications in traditional medicine for example as a remedy for asthma and diabetes, and was found to have antibacterial properties. In this work, an antioxidant and antimicrobial activities was performed on phenolic extracts of Marrubium deserti, Ammodaucus leucotrichus plants. The yield of methanol maceration of these plants is 12.4% and 20.4% respectively. The content of total polyphenols, flavonoids and anthocyanin in methanolic extracts, are varied between 19.52±1.88 and 59.24±3.45 mg/g gallic acid equivalent, and 2.08±0.29 to 1.46±0.39 mg/g quercetin equivalent, and 0.395 to 1.934µmol/g respectively. The total chlorophylls and carotenoids were be ranged from 0.149±0.20 to 1.537±0.20 g/ml and 1.537±0.20 to 0.149 ± 0.20 g/ml, respectively. According to DPPH and FRAP test, the values of EC50 was shows a higher activity of Marrubium deserti than Ammodaucus leucotrichus with EC50 values (DPPH) were 34.53±0.71 μg/mL and 258.60±15.67 mg/ml respectively. The TEAC values of FRAP test was a highly superior for Marrubium deserti 209.66±0.26 mg Equivalent Trolox/g dry residue than Ammodaucus leucotrichus 45.88±2.93 mg Trolox Equivalent/g dry residue. The antimicrobial activity against nine strains of bacteria (Staphylococcus aureus(+), Staphylococcus aureus (-), Bacillus cereus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Salmonella typhi), was showed that the tested extracts are a significant antibacterial activity with inhibition zones ranging from 10 to 50 mm. the value of CMI were ranging from 0.89 to 14.29 mg/ml.

Keywords: phenolic extract, antioxidant activity, antimicrobial activity, Marrubium deserti, Ammodaucus leucotrichus

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1008 Impact of Microbial Pathogen on Aquatic Environment

Authors: Muhammad Younis Laghari

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Global climate change has had many effects on the aquatic environment, and the major issue is pollution. Along with the other pollutants, there are a significant number of human microbial pathogens that pollute the water bodies. Another concern about the water quality is that the major aquatic resources bring water-borne pathogens and other related diseases. These resources include industrial effluent, untreated domestic sewage, acid mine drainage, etc. However, these water discharges through various routes may have treatment to eliminate the pathogenic microbes. Therefore, it is essential to control the leakage from sewer systems, residential discharge, and agricultural run-off. These pathogenic microbes have been implicated in the lives of water health (fishes), which is harmful and causes diseases. Mostly, the mortality of aquatic species results because of catastrophic floods due to poor water waste treatment and sanitation that introduce pathogenic bacteria into rivers. Pathogens survive in rivers and remain poorly known but essential to control water-borne diseases. The presence of bacteria in watercourses is diverse and constitutes a complicated subject. Many species are autochthonous and play an important role in aquatic ecosystems, while many others arise from untreated or poorly treated waste from industrial and domestic sources. Further, more investigation is required to know the induction of water-borne pathogens in various water resources and the potential impacts of water resource development on pathogen contamination.

Keywords: microbial pathogens, contamination, water resources, river water body

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1007 The Acute Impact of the Intake of Breadsticks from Different Durum Wheat Flour Mixtures on Postprandial Inflammation, Oxidative Stress, and Antiplatelet Activity in Healthy Volunteers: A Pilot Cross-Over Nutritional Intervention

Authors: O. I. Papagianni, P. Potsaki, K. Almpounioti, D. Chatzicharalampous, A. Voutsa, O. Katira, A. Michalaki, H. C. Karantonis, A. E. Koutelidakis

Abstract:

High intakes of carbohydrates and fats have been associated with an increased risk of chronic diseases due to the role of postprandial oxidative stress. This pilot nutritional intervention aimed to examine the acute effect of consuming two different types of breadsticks prepared from durum wheat flour mixtures differing in total phenolic content on postprandial inflammatory and oxidant responses in healthy volunteers. A cross-over, controlled, and single-blind clinical trial was designed, and two isocaloric high-fat and high-carbohydrate meals were tested. Serum total, HDL- and LDL-cholesterol, triglycerides, glucose, CRP, uric acid, plasma total antioxidant capacity, and antiplatelet activity were determined in fasting and 30, 60, and 120 min after consumption. The results showed a better postprandial HDL-cholesterol and total antioxidant activity response in the intervention group. The choice of durum wheat flours with higher phenolic content and antioxidant activity is presented as promising for human health, and clinical studies will expand to draw safer conclusions.

Keywords: breadsticks, durum wheat flours, postprandial inflammation, postprandial oxidative stress, ex vivo antiplatelet activity

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1006 Electrophoretic Changes in Testis and Liver of Mice after Exposure to Diclofenac Sodium

Authors: Deepak Mohan, Sushma Sharma, Mohammad Asif

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Diclofenac sodium being one of the most common non-steroidal anti-inflammatory drugs is normally used as painkiller and to reduce inflammation. The drug is known to alter the enzymatic activities of acid and alkaline phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminases. The drug also results in change in the concentration of proteins and lipids in the body. The present study is an attempt to study different biochemical changes electrophoretically due to administration of different doses of diclofenac (4mg/kg/body weight and 14mg/kg/body weight) on liver and testes of mice from 7-28 days of investigation. Homogenization of the tissue was done, supernatant separated was loaded in the gel and native polyacrylamide gel electrophoresis was conducted. Diclofenac administration resulted in alterations of all these biochemical parameters which were observed in native polyacrylamide gel electrophoretic studies. The severe degenerative changes as observed during later stages of the experiment showed correlation with increase or decrease in the activities of all the enzymes studied in the present investigation. Image analysis of gel in liver showed a decline of 7.4 and 5.3 % in low and high dose group after 7 days whereas a decline of 9.6 and 7.5% was registered after 28 days of investigation. Similar analysis for testis also showed an appreciable decline in the activity of alkaline phosphatase after 28 days. Gel analysis of serum was also performed to find a correlation in the enzymatic activities between the tissue and blood.

Keywords: diclofenac, inflammation, polyacrylamide, phosphatase

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1005 Identification of Candidate Gene for Root Development and Its Association With Plant Architecture and Yield in Cassava

Authors: Abiodun Olayinka, Daniel Dzidzienyo, Pangirayi Tongoona, Samuel Offei, Edwige Gaby Nkouaya Mbanjo, Chiedozie Egesi, Ismail Yusuf Rabbi

Abstract:

Cassava (Manihot esculenta Crantz) is a major source of starch for various industrial applications. However, the traditional cultivation and harvesting methods of cassava are labour-intensive and inefficient, limiting the supply of fresh cassava roots for industrial starch production. To achieve improved productivity and quality of fresh cassava roots through mechanized cultivation, cassava cultivars with compact plant architecture and moderate plant height are needed. Plant architecture-related traits, such as plant height, harvest index, stem diameter, branching angle, and lodging tolerance, are critical for crop productivity and suitability for mechanized cultivation. However, the genetics of cassava plant architecture remain poorly understood. This study aimed to identify the genetic bases of the relationships between plant architecture traits and productivity-related traits, particularly starch content. A panel of 453 clones developed at the International Institute of Tropical Agriculture, Nigeria, was genotyped and phenotyped for 18 plant architecture and productivity-related traits at four locations in Nigeria. A genome-wide association study (GWAS) was conducted using the phenotypic data from a panel of 453 clones and 61,238 high-quality Diversity Arrays Technology sequencing (DArTseq) derived Single Nucleotide Polymorphism (SNP) markers that are evenly distributed across the cassava genome. Five significant associations between ten SNPs and three plant architecture component traits were identified through GWAS. We found five SNPs on chromosomes 6 and 16 that were significantly associated with shoot weight, harvest index, and total yield through genome-wide association mapping. We also discovered an essential candidate gene that is co-located with peak SNPs linked to these traits in M. esculenta. A review of the cassava reference genome v7.1 revealed that the SNP on chromosome 6 is in proximity to Manes.06G101600.1, a gene that regulates endodermal differentiation and root development in plants. The findings of this study provide insights into the genetic basis of plant architecture and yield in cassava. Cassava breeders could leverage this knowledge to optimize plant architecture and yield in cassava through marker-assisted selection and targeted manipulation of the candidate gene.

Keywords: manihot esculenta crantz, plant architecture, dartseq, snp markers, genome-wide association study

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1004 Evaluation and Provenance Studies of Heavy Mineral Deposits in Recent Sediment of Ologe Lagoon, South Western, Nigeria

Authors: Mayowa Philips Ibitola, Akinade-Solomon Olorunfemi, Abe Oluwaseun Banji

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Heavy minerals studies were carried out on eighteen sediment samples from Ologe lagoon located at Lagos Barrier complex, with the aim of evaluating the heavy mineral deposits and determining the provenance of the sediments. The samples were subjected to grain analysis techniques in order to collect the finest grain size. Separation of heavy minerals from the samples was done with the aid of bromoform to enable petrographic analyses of the heavy mineral suite, under the polarising microscope. The data obtained from the heavy mineral analysis were used in preparing histograms and pie chart, from which the individual heavy mineral percentage distribution and ZTR index were derived. The percentage composition of the individual heavy mineral analyzed are opaque mineral 63.92%, Zircon 12.43%, Tourmaline 5.79%, Rutile 13.44%, Garnet 1.74% and Staurolite 3.52%. The calculated zircon, tourmaline, rutile index in percentage (ZTR) varied between 76.13 -92.15%, average garnet-zircon index (GZI), average rutile-zircon index (RuZI) and average staurolite-zircon index values in all the stations are 16.18%, 54.33%, 25.11% respectively. The mean ZTR index percentage value is 85.17% indicates that the sediments within the lagoon are mineralogically matured. The high percentage of zircon, rutile, and tourmaline indicates an acid igneous rock source for the sediments. However, the low percentage of staurolite, rutile and garnet occurrence indicates sediment of metamorphic rock source input.

Keywords: lagoon, provenance, heavy mineral, ZTR index

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1003 Improving a Stagnant River Reach Water Quality by Combining Jet Water Flow and Ultrasonic Irradiation

Authors: A. K. Tekile, I. L. Kim, J. Y. Lee

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Human activities put freshwater quality under risk, mainly due to expansion of agriculture and industries, damming, diversion and discharge of inadequately treated wastewaters. The rapid human population growth and climate change escalated the problem. External controlling actions on point and non-point pollution sources are long-term solution to manage water quality. To have a holistic approach, these mechanisms should be coupled with the in-water control strategies. The available in-lake or river methods are either costly or they have some adverse effect on the ecological system that the search for an alternative and effective solution with a reasonable balance is still going on. This study aimed at the physical and chemical water quality improvement in a stagnant Yeo-cheon River reach (Korea), which has recently shown sign of water quality problems such as scum formation and fish death. The river water quality was monitored, for the duration of three months by operating only water flow generator in the first two weeks and then ultrasonic irradiation device was coupled to the flow unit for the remaining duration of the experiment. In addition to assessing the water quality improvement, the correlation among the parameters was analyzed to explain the contribution of the ultra-sonication. Generally, the combined strategy showed localized improvement of water quality in terms of dissolved oxygen, Chlorophyll-a and dissolved reactive phosphate. At locations under limited influence of the system operation, chlorophyll-a was highly increased, but within 25 m of operation the low initial value was maintained. The inverse correlation coefficient between dissolved oxygen and chlorophyll-a decreased from 0.51 to 0.37 when ultrasonic irradiation unit was used with the flow, showing that ultrasonic treatment reduced chlorophyll-a concentration and it inhibited photosynthesis. The relationship between dissolved oxygen and reactive phosphate also indicated that influence of ultra-sonication was higher than flow on the reactive phosphate concentration. Even though flow increased turbidity by suspending sediments, ultrasonic waves canceled out the effect due to the agglomeration of suspended particles and the follow-up settling out. There has also been variation of interaction in the water column as the decrease of pH and dissolved oxygen from surface to the bottom played a role in phosphorus release into the water column. The variation of nitrogen and dissolved organic carbon concentrations showed mixed trend probably due to the complex chemical reactions subsequent to the operation. Besides, the intensive rainfall and strong wind around the end of the field trial had apparent impact on the result. The combined effect of water flow and ultrasonic irradiation was a cumulative water quality improvement and it maintained the dissolved oxygen and chlorophyll-a requirement of the river for healthy ecological interaction. However, the overall improvement of water quality is not guaranteed as effectiveness of ultrasonic technology requires long-term monitoring of water quality before, during and after treatment. Even though, the short duration of the study conducted here has limited nutrient pattern realization, the use of ultrasound at field scale to improve water quality is promising.

Keywords: stagnant, ultrasonic irradiation, water flow, water quality

Procedia PDF Downloads 177
1002 Effect of Convective Dryness Combined with Osmotic Dehydration, Blanching, Microwave and Ultrasonic Treatment on Bioactive Compounds and Rehydration Capacity of Dried Plums

Authors: Elena Corina Popescu, Magda Gabriela Bratu

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Increasing interest in keeping bioactive compounds (anthocyanins, vitamin C) and dried fruit quality has motivated the researchers to investigate new combined drying technologies. The aim of this study was to evaluate the effects of convective dryness combined with osmotic dehydration, blanching, microwave treatment and ultrasonic treatment on the quality of dried plums. Osmotic dehydration was achieved by maintaining plums for 1 h in sucrose solution (300Brix). For microwave treatment, the plums were kept at 400 W for 80 sec. For ultrasonic treatment, plums were immersed in distilled water and sonicated for 30 minutes at 40 kHz and 200 W. The blanching consists of immersing plums in hot water at 90°C for 20 seconds and cooling them rapidly. Conventional drying was carried out at 70°C for 630 minutes. Drying curves, drying rate, anthocyanin and vitamin C stability, acidity variation (expressed as malic acid), reducing sugar content, and rehydration capacity of dried plums were analyzed. Blanching led to the largest amount of evaporated water. Blanched plums have had 13.36% less water than sonicated ones. The lowest anthocyanal loss of 34.5% was obtained in osmotically dehydrated plums, and 2.93% vitamin C is found in the plums sonicated. There were no significant differences in regards acidity and reducing sugar. The plums blanched before drying have had a high capacity of rehydration.

Keywords: anthocyanin, dried plums, pretreatments, vitamin C

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1001 Comparative Study of Bread Prepared with and without Germinated Soyabean (Glycine Max) Flour

Authors: Muhammad Arsalan Mahmoo, Allah Rakha, Muhammad Sohail

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The supplementation of wheat flour with high lysine legume flours has positive effects on the nutritional value of bread. In present study, germinated and terminated soya flour blends were prepared and supplemented in bread in variable proportions (10 % and 20 % of each) to check its impact on quality and sensory attributes of bread. The results showed that there was a significant increase in protein, ash and crude fat contents due to increase in the level of germinated and ungerminated soya flour. However, the moisture and crude fiber contents of composite flours containing germinated and ungerminated soya flour decreased with increased level of supplementation. Mean values for physical analysis (loaf volume, specific volume, weight loss and force for texture) were significantly higher in breads prepared with germinated soya bean flour.The scores assigned to sensory parameters of breads like volume, color of crust, symmetry, color of crumb, texture, taste and aroma decreased significantly by increasing the level of germinated and ungerminated soya flour in wheat flour while color of crust and taste slightly improved. The scores given to overall acceptability of bread prepared from composite flour supplemented with 10 % germinated soya flour.

Keywords: composite bread, protein energy malnutrition, supplementation, amino acid profile, grain legumes

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1000 Phosphate Capture from Sewage by Hafnium-Modified Fe₃O₄@SiO₂ Superparamagnetic Nanoparticles: Adsorption Capacity, Selectivity, Reusability Analysis and Mechanistic Insights

Authors: Qian Zhao

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With global increasing demand for phosphorus and intensively depleting reserves, it is urgent need to explore innovative approaches towards capturing phosphate from sewage, which is also an effective way to reduce phosphate contamination and avoid eutrophication of water bodies. In the present article, the superparamagnetic nano-sorbents containing Fe₃O₄ core and hafnium-modified MgAl/MgFe layered double hydroxides shell (abbreviated as MgAlHf-NP and MgFeHf-NP) was developed using a simple and low-cost synthesis protocol. The obtained Hf-coated nano-materials showed well-defined crystal structure and sufficient saturation magnetization and exhibited higher adsorption capacity for phosphate. Meanwhile, high selectivity was also confirmed since coexisting foreign anions and biomacromolecules showed little competitive effect on phosphate adsorption. The enhancement via doping with Hf should be explained by the stronger ligand complexation built by the pair of hard acid Hf ion and hard base phosphate that matched up the bonding preferences. Sufficient OH⁻ concentration and clear pH shift during the desorption/regeneration allowed for regeneration rate of higher than 90% after 5 cycles of adsorption desorption. This article attempts to provide a competitive candidate for phosphate-capture, which is highly effective, easily separable and repeatedly usable.

Keywords: phosphate recovery, nanoparticles, superparamagnetic, adsorption, reusability

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999 Optimal and Critical Path Analysis of State Transportation Network Using Neo4J

Authors: Pallavi Bhogaram, Xiaolong Wu, Min He, Onyedikachi Okenwa

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A transportation network is a realization of a spatial network, describing a structure which permits either vehicular movement or flow of some commodity. Examples include road networks, railways, air routes, pipelines, and many more. The transportation network plays a vital role in maintaining the vigor of the nation’s economy. Hence, ensuring the network stays resilient all the time, especially in the face of challenges such as heavy traffic loads and large scale natural disasters, is of utmost importance. In this paper, we used the Neo4j application to develop the graph. Neo4j is the world's leading open-source, NoSQL, a native graph database that implements an ACID-compliant transactional backend to applications. The Southern California network model is developed using the Neo4j application and obtained the most critical and optimal nodes and paths in the network using centrality algorithms. The edge betweenness centrality algorithm calculates the critical or optimal paths using Yen's k-shortest paths algorithm, and the node betweenness centrality algorithm calculates the amount of influence a node has over the network. The preliminary study results confirm that the Neo4j application can be a suitable tool to study the important nodes and the critical paths for the major congested metropolitan area.

Keywords: critical path, transportation network, connectivity reliability, network model, Neo4j application, edge betweenness centrality index

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998 Controlled Drug Delivery System for Delivery of Poor Water Soluble Drugs

Authors: Raj Kumar, Prem Felix Siril

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The poor aqueous solubility of many pharmaceutical drugs and potential drug candidates is a big challenge in drug development. Nanoformulation of such candidates is one of the major solutions for the delivery of such drugs. We initially developed the evaporation assisted solvent-antisolvent interaction (EASAI) method. EASAI method is use full to prepared nanoparticles of poor water soluble drugs with spherical morphology and particles size below 100 nm. However, to further improve the effect formulation to reduce number of dose and side effect it is important to control the delivery of drugs. However, many drug delivery systems are available. Among the many nano-drug carrier systems, solid lipid nanoparticles (SLNs) have many advantages over the others such as high biocompatibility, stability, non-toxicity and ability to achieve controlled release of drugs and drug targeting. SLNs can be administered through all existing routes due to high biocompatibility of lipids. SLNs are usually composed of lipid, surfactant and drug were encapsulated in lipid matrix. A number of non-steroidal anti-inflammatory drugs (NSAIDs) have poor bioavailability resulting from their poor aqueous solubility. In the present work, SLNs loaded with NSAIDs such as Nabumetone (NBT), Ketoprofen (KP) and Ibuprofen (IBP) were successfully prepared using different lipids and surfactants. We studied and optimized experimental parameters using a number of lipids, surfactants and NSAIDs. The effect of different experimental parameters such as lipid to surfactant ratio, volume of water, temperature, drug concentration and sonication time on the particles size of SLNs during the preparation using hot-melt sonication was studied. It was found that particles size was directly proportional to drug concentration and inversely proportional to surfactant concentration, volume of water added and temperature of water. SLNs prepared at optimized condition were characterized thoroughly by using different techniques such as dynamic light scattering (DLS), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), atomic force microscopy (AFM), X-ray diffraction (XRD) and differential scanning calorimetry and Fourier transform infrared spectroscopy (FTIR). We successfully prepared the SLN of below 220 nm using different lipids and surfactants combination. The drugs KP, NBT and IBP showed 74%, 69% and 53% percentage of entrapment efficiency with drug loading of 2%, 7% and 6% respectively in SLNs of Campul GMS 50K and Gelucire 50/13. In-vitro drug release profile of drug loaded SLNs is shown that nearly 100% of drug was release in 6 h.

Keywords: nanoparticles, delivery, solid lipid nanoparticles, hot-melt sonication, poor water soluble drugs, solubility, bioavailability

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997 The Genetic Basis of the Lack of Impulse Control: What is Provided for the Criminal Law?

Authors: Amir Bastani

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The result of the research in the field of human behavioural genetics demonstrates a genetic contribution of behavioural differences in aggression, violence, drug and substance abuse, antisocial personality disorder and other related traits. As the field of human behavioural genetics progresses and achieves credibility, the criminal accused continue to use its types of evidence into the criminal law. One of the most important genetic factors which controls certain neurotransmitters like dopamine and serotonin is the Monoamine Oxidase Acid A (MAOA) gene, known as the 'warrior gene'. The high-profile study by Caspi and colleagues in 2002 showed that the combination between one type of variation of the MAOA gene and childhood maltreatment noticeably predisposes a person to antisocial behaviour. Moreover, further scientific research shows that individuals with the MAOA gene have to some degree difficulties in controlling their impulses. Based on the evidence of MAOA, some criminal accused claimed difficulties in self-control. In the first case – the famous case of Mobley – the court rejected the MAOA evidence on the ground of the lack of scientific support. In contrast, in other cases after the Mobley trial, courts accepted the evidence of MAOA. In this paper, the issue of lack of impulse control produced by the MAOA gene and cases which relied on the MAOA evidence and successfully being accepted will be reviewed in detail. Finally, the anticipation of the paper for the future use of the MAOA evidence in criminal cases will be presented.

Keywords: genetic defence, criminal responsibility, MAOA, self-control

Procedia PDF Downloads 452
996 The Historical Background of Physical Changing Towards Ancient Mosques in Aceh, Indonesia

Authors: Karima Adilla

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Aceh province, into which Islam convinced to have entered Indonesia in the 12th Century before spreading throughout the archipelago and the rest of Southeast Asia, has several early Islamic mosques that still exist until today. However, due to some circumstances, the restoration and rehabilitation towards those mosques have been made in some periods, while the background was diverse. Concerning this, the research will examine the physical changing aspects of 3 prominent historical mosques in Aceh Besar and Banda Aceh; those are, Indrapuri Mosque, Baiturrahman Grand Mosque, and Baiturrahim Mosque built coincided with the beginning of Islam’s development in Aceh and regarded as eventful mosques. The existence of Indrapuri Mosque built on the remains of the Lamuri Kingdom’s temple is a historical trace that there was Hindu-Buddhist civilization in Aceh before Islam entered and became the majority religion about 98% from Aceh total population. Also, there was the Dutch who colonialized Aceh behind the existence of two famous mosques in Aceh, namely Baiturrahman Grand Mosque and Baiturrahim Mosque, as the colonizer also assisted to rebuild those 2 sacred Mosques to quell the anger of the Acehnese people because their mosque was burnt by the Dutch. Interestingly, despite underwent a long history successively since the rise of Islam after the Hindu-Buddhist kingdom had collapsed, colonialization, conflict, in Aceh, and even experienced the earthquake and tsunami disaster in 2004, those mosques still exist. Therefore, those mosques have been considered as historical silent witnesses. However, it was not merely those reasons that led the mosques underwent several physical changes, otherwise economic, political, social, cultural and religious factors were also highly influential. Instead of directly illustrating the physical changing of those three mosques, this research intends to identify under what condition the physical appearance continuously changing during the sultanate era, the colonial period until post-independent in terms of the architectural style, detail elements, design philosophy, and how the remnants buildings act as medium to bridge the history. A framework will use qualitative research methods by collecting actual data of the mosque's physical change figures through field studies, investigations, library studies and interviews. This research aims to define every trace of historical issues embedded in the physical changing of those mosques as they are intertwined in collecting historical proof. Thus, the result will reveal the characteristic interrelation between history, the mosque architectural style in a certain period, the physical changes background and its impact. Eventually, this research will also explicate a clear inference of each mosque’s role in representing history in Aceh Besar and Banda Aceh specifically, as well as Aceh generally through architectural design concepts.

Keywords: Aceh ancient mosques, Aceh history, Islamic architecture, physical changing

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995 Quantitative Analysis of Caffeine in Pharmaceutical Formulations Using a Cost-Effective Electrochemical Sensor

Authors: Y. T. Gebreslassie, Abrha Tadesse, R. C. Saini, Rishi Pal

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Caffeine, known chemically as 3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-dione, is a naturally occurring alkaloid classified as an N-methyl derivative of xanthine. Given its widespread use in coffee and other caffeine-containing products, it is the most commonly consumed psychoactive substance in everyday human life. This research aimed to develop a cost-effective, sensitive, and easily manufacturable sensor for the detection of caffeine. Antraquinone-modified carbon paste electrode (AQMCPE) was fabricated, and the electrochemical behavior of caffeine on this electrode was investigated using cyclic voltammetry (CV) and square wave voltammetry (SWV) in a solution of 0.1M perchloric acid at pH 0.56. The modified electrode displayed enhanced electrocatalytic activity towards caffeine oxidation, exhibiting a two-fold increase in peak current and an 82 mV shift of the peak potential in the negative direction compared to an unmodified carbon paste electrode (UMCPE). Exploiting the electrocatalytic properties of the modified electrode, SWV was employed for the quantitative determination of caffeine. Under optimized experimental conditions, a linear relationship between peak current and concentration was observed within the range of 2.0 x 10⁻⁶ to 1.0× 10⁻⁴ M, with a correlation coefficient of 0.998 and a detection limit of 1.47× 10⁻⁷ M (signal-to-noise ratio = 3). Finally, the proposed method was successfully applied to the quantitative analysis of caffeine in pharmaceutical formulations, yielding recovery percentages ranging from 95.27% to 106.75%.

Keywords: antraquinone-modified carbon paste electrode, caffeine, detection, electrochemical sensor, quantitative analysis

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994 Optimization of Hepatitis B Surface Antigen Purifications to Improving the Production of Hepatitis B Vaccines on Pichia pastoris

Authors: Rizky Kusuma Cahyani

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Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

Procedia PDF Downloads 130
993 Targeting Tumour Survival and Angiogenic Migration after Radiosensitization with an Estrone Analogue in an in vitro Bone Metastasis Model

Authors: Jolene M. Helena, Annie M. Joubert, Peace Mabeta, Magdalena Coetzee, Roy Lakier, Anne E. Mercier

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Targeting the distant tumour and its microenvironment whilst preserving bone density is important in improving the outcomes of patients with bone metastases. 2-Ethyl-3-O-sulphamoyl-estra1,3,5(10)16-tetraene (ESE-16) is an in-silico-designed 2- methoxyestradiol analogue which aimed at enhancing the parent compound’s cytotoxicity and providing a more favourable pharmacokinetic profile. In this study, the potential radiosensitization effects of ESE-16 were investigated in an in vitro bone metastasis model consisting of murine pre-osteoblastic (MC3T3-E1) and pre-osteoclastic (RAW 264.7) bone cells, metastatic prostate (DU 145) and breast (MDA-MB-231) cancer cells, as well as human umbilical vein endothelial cells (HUVECs). Cytotoxicity studies were conducted on all cell lines via spectrophotometric quantification of 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide. The experimental set-up consisted of flow cytometric analysis of cell cycle progression and apoptosis detection (Annexin V-fluorescein isothiocyanate) to determine the lowest ESE-16 and radiation doses to induce apoptosis and significantly reduce cell viability. Subsequent experiments entailed a 24-hour low-dose ESE-16-exposure followed by a single dose of radiation. Termination proceeded 2, 24 or 48 hours thereafter. The effect of the combination treatment was investigated on osteoclasts via tartrate-resistant acid phosphatase (TRAP) activity- and actin ring formation assays. Tumour cell experiments included investigation of mitotic indices via haematoxylin and eosin staining; pro-apoptotic signalling via spectrophotometric quantification of caspase 3; deoxyribonucleic acid (DNA) damage via micronuclei analysis and histone H2A.X phosphorylation (γ-H2A.X); and Western blot analyses of bone morphogenetic protein-7 and matrix metalloproteinase-9. HUVEC experiments included flow cytometric quantification of cell cycle progression and free radical production; fluorescent examination of cytoskeletal morphology; invasion and migration studies on an xCELLigence platform; and Western blot analyses of hypoxia-inducible factor 1-alpha and vascular endothelial growth factor receptor 1 and 2. Tumour cells yielded half-maximal growth inhibitory concentration (GI50) values in the nanomolar range. ESE-16 concentrations of 235 nM (DU 145) and 176 nM (MDA-MB-231) and a radiation dose of 4 Gy were found to be significant in cell cycle and apoptosis experiments. Bone and endothelial cells were exposed to the same doses as DU 145 cells. Cytotoxicity studies on bone cells reported that RAW 264.7 cells were more sensitive to the combination treatment than MC3T3-E1 cells. Mature osteoclasts were more sensitive than pre-osteoclasts with respect to TRAP activity. However, actin ring morphology was retained. The mitotic arrest was evident in tumour and endothelial cells in the mitotic index and cell cycle experiments. Increased caspase 3 activity and superoxide production indicated pro-apoptotic signalling in tumour and endothelial cells. Increased micronuclei numbers and γ-H2A.X foci indicated increased DNA damage in tumour cells. Compromised actin and tubulin morphologies and decreased invasion and migration were observed in endothelial cells. Western blot analyses revealed reduced metastatic and angiogenic signalling. ESE-16-induced radiosensitization inhibits metastatic signalling and tumour cell survival whilst preferentially preserving bone cells. This low-dose combination treatment strategy may promote the quality of life of patients with metastatic bone disease. Future studies will include 3-dimensional in-vitro and murine in-vivo models.

Keywords: angiogenesis, apoptosis, bone metastasis, cancer, cell migration, cytoskeleton, DNA damage, ESE-16, radiosensitization.

Procedia PDF Downloads 142