Search results for: methyl eugenol
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 477

Search results for: methyl eugenol

177 Targeting Calcium Dysregulation for Treatment of Dementia in Alzheimer's Disease

Authors: Huafeng Wei

Abstract:

Dementia in Alzheimer’s Disease (AD) is the number one cause of dementia internationally, without effective treatments. Increasing evidence suggest that disruption of intracellular calcium homeostasis, primarily pathological elevation of cytosol and mitochondria but reduction of endoplasmic reticulum (ER) calcium concentrations, play critical upstream roles on multiple pathologies and associated neurodegeneration, impaired neurogenesis, synapse, and cognitive dysfunction in various AD preclinical studies. The last federal drug agency (FDA) approved drug for AD dementia treatment, memantine, exert its therapeutic effects by ameliorating N-methyl-D-aspartate (NMDA) glutamate receptor overactivation and subsequent calcium dysregulation. More research works are needed to develop other drugs targeting calcium dysregulation at multiple pharmacological acting sites for future effective AD dementia treatment. Particularly, calcium channel blockers for the treatment of hypertension and dantrolene for the treatment of muscle spasm and malignant hyperthermia can be repurposed for this purpose. In our own research work, intranasal administration of dantrolene significantly increased its brain concentrations and durations, rendering it a more effective therapeutic drug with less side effects for chronic AD dementia treatment. This review summarizesthe progress of various studies repurposing drugs targeting calcium dysregulation for future effective AD dementia treatment as potentially disease-modifying drugs.

Keywords: alzheimer, calcium, cognitive dysfunction, dementia, neurodegeneration, neurogenesis

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176 Layer by Layer Coating of Zinc Oxide/Metal Organic Framework Nanocomposite on Ceramic Support for Solvent/Solvent Separation Using Pervaporation Method

Authors: S. A. A. Nabeela Nasreen, S. Sundarrajan, S. A. Syed Nizar, Seeram Ramakrishna

Abstract:

Metal-organic frameworks (MOFs) have attracted considerable interest due to its diverse pore size tunability, fascinating topologies and extensive uses in fields such as catalysis, membrane separation, chemical sensing, etc. Zeolitic imidazolate frameworks (ZIFs) are a class of MOF with porous crystals containing extended three-dimensional structures of tetrahedral metal ions (e.g., Zn) bridged by Imidazolate (Im). Selected ZIFs are used to separate solvent/solvent mixtures. A layer by layer formation of the nanocomposite of Zinc oxide (ZnO) and ZIF on a ceramic support using a solvothermal method was engaged and tested for target solvent/solvent separation. Metal oxide layer was characterized by XRD, SEM, and TEM to confirm the smooth and continuous coating for the separation process. The chemical composition of ZIF films was studied by using X-Ray absorption near-edge structure (XANES) spectroscopy. The obtained ceramic tube with metal oxide and ZIF layer coating were tested for its packing density, thickness, distribution of seed layers and variation of permeation rate of solvent mixture (isopropyl alcohol (IPA)/methyl isobutyl ketone (MIBK). Pervaporation technique was used for the separation to achieve a high permeation rate with separation ratio of > 99.5% of the solvent mixture.

Keywords: metal oxide, membrane, pervaporation, solvothermal, ZIF

Procedia PDF Downloads 194
175 Comparative Antibacterial Property of Matured Trunk and Stem Bark Extract of Tamarindus indica L., Preformulation, Development and Quality Control of Cream

Authors: A. M. T. Jacinto, M.O. Osi

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Tamarind has various medicinal properties among which is its antibacterial property. Its bark contains saponins, alkaloids, sesquiterpenes and tannins. It is rich in phlobapenes which is responsible for antibacterial property. The objective of the study was to determine which bark will produce the highest antibacterial property, develop it into a topical cream and evaluate its quality and characteristics. Powdered barks of Tamarind were extracted by soxhlet method using 70% acetone. Stem bark produced a higher yield than trunk bark (5.85 g vs. 4.73 g). It was found that the trunk bark was more sensitive than stem bark to microorganisms namely Staphylococcus aureus, Corynebacterium minutissimum, and Streptococcus spp. Sensitivity of trunk bark can be attributed to a more developed phytoconstituents. Dermal sensitization test on both sexes of rabbits using the following concentrations: 100%, 40% and 20% of extract showed that Tamarind has no irritating property and therefore safe for formulation into an antibacterial cream. Excipients used for formulation such as methyl paraben, propyl paraben, stearyl alcohol and white petrolatum were compatible with the Tamarind acetone extract through Differential Scanning Calorimetry except sodium lauryl sulfate that exhibited crystallization when subjected at 200˚C. The method of manufacture used in cream is fusion, therefore strict compliance of processing temperature should be observed to prevent polymorphism. Quality control tests of formulated cream based on USP 30 and Philippine Pharmacopeia were satisfactory.

Keywords: antibacterial, differential scanning calorimetry, tannins, dermal sensitization

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174 Synthesis and Characterization of New Thermotropic Monomers – Containing Phosphorus

Authors: Diana Serbezeanu, Ionela-Daniela Carja, Tachita Vlad-Bubulac, Sergiu Sova

Abstract:

New phosphorus-containing monomers having methoxy end functional groups were prepared from methyl 4-hydroxybenzoate and two different dichlorides with phosphorus, namely phenyl phosphonic dichloride and phenyl dichlorophosphate. The structures of the monomers were confirmed by FTIR and NMR spectroscopy. The assignments for the 1H, 13C and 31P chemical shifts are based on 1D and 2D NMR homo- and heteronuclear correlations (H,H-COSY (Correlation Spectroscopy), H,C-HMQC (Heteronuclear Multiple Quantum Correlation and H,C-HMBC (Heteronuclear Multiple Bond Correlation)) and 31P-13C couplings. The monomers exhibited good solubility in common organic solvents. Dimethyl sulfoxide was to be a good solvent to grow crystals of considerable size which were investigated by X-ray analysis. One of these two new monomers presented thermotropic liquid crystalline behaviour, as revealed by differential scanning calorimetry (DSC), polarized light microscopy (PLM) and X-ray diffraction (XRD). The transition temperature from crystal to liquid crystalline state (K→LC) was 143°C and from the LC to isotropic state (LC→I) was 167°C. Upon heating, bis(4-(methoxycarbonyl)phenyl formed fine textures, difficult to be ascribed to smectic or nematic phases. Upon cooling from the isotropic state, bis(4-(methoxycarbonyl)phenyl exhibited a mosaic-type texture. X-ray diffraction measurements at small angles (SAXS) of bis(4-(methoxycarbonyl)phenyl showed two peaks at 1.8 Å and 3.5 Å, respectively suggesting organization at supramolecular level.

Keywords: phosphorus-containing monomers, polarized light microscopy, structure investigation, thermotropic liquid crystalline properties

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173 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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172 Biobased Toughening Filler for Polylactic Acid from Ultrafine Fully Vulcanized Powder Natural Rubber Grafted with Polymethylmethacrylate

Authors: Panyawutthi Rimdusit, Krittapas Charoensuk, Sarawut Rimdusit

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A biobased toughening filler for polylactic acid (PLA) based on natural rubber is developed in this work. Deproteinized natural rubber (DPNR) was modified by grafting polymerization with methyl methacrylate monomer (MMA) and further crosslinked by e-beam irradiation and spray drying process to achieve ultrafine full vulcanized powdered natural rubber grafted with polymethylmethacrylate (UFPNRg-PMMA) to solves in the challenges of incompatibility between natural rubber and PLA. Intriguingly, UFPNR-g-PMMA revealed outstanding and unique properties with minimal particle aggregation. The average particle size of rubber powder obtained from UFPNR-g-PMMA at PMMA grafting content of 20 phr reduced to 3.3±1.2 µm, compared to that of neat UFPNR of 5.3±2.3 µm which also showed partial particle aggregation. It is also found that the impact strength of the filled PLA was enhanced to 33.4±5.6 kJ/m2 at PLA/UFPNR-gPMMA 20 wt% compared to neat PLA of 9.6±3 kJ/m2. The thermal degradation temperature of the PLA composites was enhanced with increasing UFPNR-g-PMMA content without affecting the glass transition temperature of the composites. The fracture surface of PLA/ UFPNR-g-PMMA suggested internal cavitation and crazes are the main effects of rubber toughening PLA with substantial interfacial interaction between the filler and the matrix.

Keywords: natural rubber, ultrafine fully vulcanized powder rubber, polylactic acid, polymer composites

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171 Modifiable Poly Methacrylic Acid-Co-Acrylonitrile Microgels Fabricated with Cu and Co Nanoparticles for Simultaneous Catalytic Reduction of Multiple Compounds

Authors: Muhammad Ajmal, Muhammad Siddiq, Nurettin Sahiner

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We prepared poly(methacrylic acid-co-acrylonitrile) (p(MAc-co-AN)) microgels by inverse suspension polymerization, and converted the nitrile groups into amidoxime groups to obtain more hydrophilic amidoximated poly(methacrylic acid-co-acrylonitile) (amid-p(MAc-co-AN)) microgels. Amid-microgels were used as microreactors for in situ synthesis of copper and cobalt nanoparticles. Cu (II) and Co (II) ions were loaded into microgels from their aqueous metal salt solutions and then converted to corresponding metal nanoparticle (MNP) by treating the loaded metal ions with sodium borohydride (NaBH4). The characterization of the prepared microgels and microgel metal nanoparticle composites was carried out by SEM, TEM and TG analysis. The amounts of metal nanoparticles within microgels were estimated by AAS measurements by dissolving the MNP entrapped within microgels by concentrated HCl acid treatment. Catalytic performances of the prepared amid-p(MAc-co-AN)-M (M: Cu, Co) microgel composites were investigated by using them as catalyst for the degradation of cationic and anionic organic dyes such as eosin Y (EY), methylene blue (MB) and methyl Orange (MO), and for the reduction of nitro aromatic pollutants like 2-nitrophenol (2-NP) and 4-nitrophenol (4-NP) to their corresponding amino phenols. Here, we also report for the first time, the simultaneous degradation/reduction of MB, EY, and 4-NP by amid-p(MAc-co-AN)-Cu microgel composites. Different parameters affecting the reduction rates such as metal types, amount of catalysts, temperature and the amount of reducing agent were investigated.

Keywords: microgels, nanoparticles, catalyst, pollutants

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170 Inhibition of Streptococcus Mutans Biofilm Development of Dental Caries In Vitro and In Vivo by Trachyspermum ammi Seeds: An Approach of Alternative Medicine

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

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

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

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169 Silver Grating for Strong and Reproducible SERS Response

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

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

Keywords: grating, nanostructures, plasmon-polaritons, SERS

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168 Microbiological Analysis of Biofuels in Order to Follow Stability on Room Temperature

Authors: Radovan Cobanovic, Milica Rankov Sicar

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Biodiesel refers to a vegetable oil - or animal fat-based diesel fuel consisting of long-chain alkyl (methyl, ethyl, or propyl) esters. It is derived by alcoholysis of triacylglycerols (triglycerides) from various lipid based materials that can be traditionally categorized into the following main groups: vegetable oils, animal fats, waste and algal oils. The goal of this study was to evaluate microbiological stability of biodiesel samples since it has been made from vegetable oil or animal fat which was stored on room temperature. For the purposes of this study, analyzes were conducted on six samples of biodiesel first at zero sample at the reception day than fifth, thirtieth, sixtieth, ninetieth and one hundred twentieth day from the day of reception. During this period, biodiesel samples were subjected to microbiological analyses (Salmonella spp., Listeria monocytogenes, Enterobacteriaceae and total plate count). All analyses were tested according to ISO methodology: Salmonella spp ISO 6579, Listeria monocytogenes ISO 11290-2, Enterobacteriaceae ISO 21528-1, total plate count ISO 4833-1. The results obtained after the analyses which were done according to the plan during the 120 days indicate that are no changes of products concerning microbiological analyses. Salmonella spp., Listeria monocytogenes, Enterobacteriaceae were not detected and results for total plate count showed values < 10 cfu/g for all six samples. On the basis of this monitoring under defined storage conditions at room temperatures, the results showed that biodiesel is very stable as far as microbiological analysis were concerned.

Keywords: biodiesel, microbiology, room temperature, stability

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167 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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166 Construction and Performance of Nanocomposite-Based Electrochemical Biosensor

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

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

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

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165 Relationship between Response of the Resistive Sensors on the Chosen Volatile Organic Compounds (VOCs) and Their Concentration

Authors: Marek Gancarz, Agnieszka Nawrocka, Robert Rusinek, Marcin Tadla

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Volatile organic compounds (VOCs) are the fungi metabolites in the gaseous form produced during improper storage of agricultural commodities (e.g. grain, food). The spoilt commodities produce a wide range of VOCs including alcohols, esters, aldehydes, ketones, alkanes, alkenes, furans, phenols etc. The characteristic VOCs and odours can be determined by using electronic nose (e-Nose) which contains a matrix of different kinds of sensors e.g. resistive sensors. The aim of the present studies was to determine relationship between response of the resistive sensors on the chosen volatiles and their concentration. According to the literature, it was chosen volatiles characteristic for the cereals: ethanol, 3-methyl-1-butanol and hexanal. Analysis of the sensor signals shows that a signal shape is different for the different substances. Moreover, each VOC signal gives information about a maximum of the normalized sensor response (R/Rmax), an impregnation time (tIM) and a cleaning time at half maximum of R/Rmax (tCL). These three parameters can be regarded as a ‘VOC fingerprint’. Seven resistive sensors (TGS2600-B00, TGS2602-B00, TGS2610-C00, TGS2611-C00, TGS2611-E00, TGS2612-D00, TGS2620-C00) produced by Figaro USA Inc., and one (AS-MLV-P2) produced by AMS AG, Austria were used. Two out of seven sensors (TGS2611-E00, TGS2612-D00) did not react to the chosen VOCs. The most responsive sensor was AS-MLV-P2. The research was supported by the National Centre for Research and Development (NCBR), Grant No. PBS2/A8/22/2013.

Keywords: agricultural commodities, organic compounds, resistive sensors, volatile

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164 Antidepressant-Like Effects of EQC-34, a 5HT3 Receptor Antagonist in Neurobehavioral Mouse Model of Depression

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

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

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

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163 Design, Synthesis and Anti-Inflammatory Activity of Some Coumarin and Flavone Derivatives Containing 1,4 Dioxane Ring System

Authors: Asif Husain, Shah Alam Khan

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Coumarins and flavones are oxygen containing heterocyclic compounds which are present in various biologically active compounds. Both the heterocyclic rings are associated with diverse biological actions, therefore considered as an important scaffold for the design of molecules of pharmaceutical interest. Aim: To synthesize and evaluate the in vivo anti-inflammatory activity of few coumrain and flavone derivatives containing 1,4 dioxane ring system. Materials and methods: Coumarin derivatives (3a-d) were synthesized by reacting 7,8 dihydroxy coumarin (2a) and its 4-methyl derivative (2b) with epichlorohydrin/ethylene dibromide. The flavone derivatives (10a-d) were prepared by using quercetin and 3,4 dihydroxy flavones. Compounds of both the series were also evaluated for their anti-inflammatory, analgesic activity and ulcerogenicity in animal models by reported methods. Results and Discussion: The structures of all newly synthesized compounds were confirmed with the help of IR, 1H NMR, 13C NMR and Mass spectral studies. Elemental analyses data for each element analyzed (C, H, N) was found to be within acceptable range of ±0.4 %. Flavone derivatives, but in particular quercetin containing 1,4 dioxane ring system (10d) showed better anti-inflammatory and analgesic activity along with reduced gastrointestinal toxicity as compared to other synthesized compounds. The results of anti-inflammatory and analgesic activities of both the series are comparable with the positive control, diclofenac. Conclusion: Compound 10d, a quercetin derivative, emerged as a lead molecule which exhibited potent anti-inflammatory and analgesic activity with significant reduced gastric toxicity.

Keywords: analgesic, anti-inflammatory, 1, 4 dioxane, coumarin, flavone

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162 Ultrasonic Investigation as Tool for Study of Molecular Interaction of 2-Hydroxy Substituted Pyrimidine Derivative at Different Concentrations

Authors: Shradha S. Binani, P. S. Bodke, R. V. Joat

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Recent decades have witnessed an exponential growth in the field of acoustical parameters and ultrasound on solid, liquid and gases. Ultrasonic propagation parameters yield valuable information regarding the behavior of liquid systems because intra and intermolecular association, dipolar interaction, complex formation and related structural changes affecting the compressibility of the system which in turn produces variations in the ultrasonic velocity. The acoustic and thermo dynamical parameters obtained in ultrasonic study show that ion-solvation is accompanied by the destruction or enhancement of the solvent structure. In the present paper the ultrasonic velocity (v), density (ρ), viscosity(η) have been measured for the pharmacological important compound 2-hydroxy substituted phenyl pyrimidine derivative (2-hydroxy-4-(4’-methoxy phenyl)-6-(2’-hydroxy-4’-methyl-5’-chlorophenyl)pyrimidine) in ethanol as a solvent by using different concentration at constant room temperature. These experimental data have been used to estimate physical parameter like adiabatic compressibility, intermolecular free length, relaxation time, free volume, specific acoustic impedance, relative association, Wada’s constant, Rao’s constant etc. The above parameters provide information in understanding the structural and molecular interaction between solute-solvent in the drug solution with respect to change in concentration.

Keywords: acoustical parameters, ultrasonic velocity, density, viscosity, 2-hydroxy substituted phenyl pyrimidine derivative

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161 Applying Cationic Porphyrin Derivative 5, 10-Dihexyl-15, 20bis Porphyrin, as Transfection Reagent for Gene Delivery into Mammalian Cells

Authors: Hajar Hosseini Khorami

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Porphyrins are organic, aromatic compounds found in heme, cytochrome, cobalamin, chlorophyll , and many other natural products with essential roles in biological processes that their cationic forms have been used as groups of favorable non-viral vectors recently. Cationic porphyrins are self-chromogenic reagents with a high capacity for modifications, great interaction with DNA and protection of DNA from nuclease during delivery of it into a cell with low toxicity. In order to have high efficient gene transfection into the cell while causing low toxicity, genetically manipulations of the non-viral vector, cationic porphyrin, would be useful. In this study newly modified cationic porphyrin derivative, 5, 10-dihexyl-15, 20bis (N-methyl-4-pyridyl) porphyrin was applied. Cytotoxicity of synthesized cationic porphyrin on Chinese Hamster Ovarian (CHO) cells was evaluated by using MTT assay. This cationic derivative is dose-dependent, with low cytotoxicity at the ranges from 100 μM to 0.01μM. It was uptake by cells at high concentration. Using direct non-viral gene transfection method and different concentration of cationic porphyrin were tested on transfection of CHO cells by applying derived transfection reagent with X-tremeGENE HP DNA as a positive control. However, no transfection observed by porphyrin derivative and the parameters tested except for positive control. Results of this study suggested that applying different protocol, and also trying other concentration of cationic porphyrins and DNA for forming a strong complex would increase the possibility of efficient gene transfection by using cationic porphyrins.

Keywords: cationic porphyrins, gene delivery, non-viral vectors, transfection reagents

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160 Estimated Human Absorbed Dose of 111 In-BPAMD as a New Bone-Seeking Spect-Imaging Agent

Authors: H. Yousefnia, S. Zolghadri

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An early diagnosis of bone metastases is very important for providing a profound decision on a subsequent therapy. A prerequisite for the clinical application of new diagnostic radiopharmaceutical is the measurement of organ radiation exposure dose from biodistribution data in animals. In this study, the dosimetric studies of a novel agent for SPECT-imaging of bone methastases, 111In-(4-{[(bis(phosphonomethyl))carbamoyl]methyl}-7,10-bis(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl) acetic acid (111In-BPAMD) complex, have been estimated in human organs based on mice data. The radiolabeled complex was prepared with high radiochemical purity at the optimal conditions. Biodistribution studies of the complex were investigated in male Syrian mice at selected times after injection (2, 4, 24 and 48 h). The human absorbed dose estimation of the complex was performed based on mice data by the radiation absorbed dose assessment resource (RADAR) method. 111In-BPAMD complex was prepared with high radiochemical purity >95% (ITLC) and specific activities of 2.85 TBq/mmol. Total body effective absorbed dose for 111In-BPAMD was 0.205 mSv/MBq. This value is comparable to the other 111In clinically used complexes. The results show that the dose to critical organs the complex is well within the acceptable considered range for diagnostic nuclear medicine procedures. Generally, 111In-BPAMD has interesting characteristics and can be considered as a viable agent for SPECT-imaging of the bone metastases in the near future.

Keywords: In-111, BPAMD, absorbed dose, RADAR

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159 Integrated Management of Tithonia Diversifolia in the Vhembe Biosphere Reserve

Authors: Mutavhatsindi Tshinakaho

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Invasive alien plants (IAP’s) are referred to as species that are non-native to the ecosystem under consideration. Whose introduction causes or is likely to cause economic, ecological, or environmental harm. The integrated management of the invasive plant, Tithonia diversifolia, will be assessed through two herbicide trials (one on the seedlings and the other on matured plants) and a competitive trial between Tithonia and invasive grass species. The initial herbicide trial will be undertaken at the University of Venda Agricultural greenhouse facilities, where Tithonia will be planted in pot plants and watered every after two days until they reach at least 30 cm and will then be subjected to four different herbicide treatments (Metsulfuron methyl, Fluroxypyr, Picloram, Triclopyr), water will be utilised as a control. The percentage damage to foliar will be recorded. The second herbicide trial will be undertaken at Levubu road site, where matured Tithonia will be cut at at least 10cm above the ground and the subjected to herbicide treatments (Picloram, Fluroxypyr, Imazapyr, and Water as a control). The site will be visited post treatment for assessment. For the competition trial, tall grass species will be chosen as competitors (Panicum maximum and Eragrostis murvula), they will be grown at six densities per pot in the greenhouse facilities at the University of Venda, were they will be kept watered for the duration of the experiment. At the end of the experiment, plants will be removed from pots, and the above and below ground biomass will be weighed. The expected results are to know the effective integrated management strategy for T. diversifolia, the effective rehabilitation of T. diversifolia invaded habitats, and the effective chemical control of T. diversifolia

Keywords: foliar, biomass, competition, invasion

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158 Development and Characterization of a Film Based on Hydroxypropyl Methyl Cellulose Incorporated by a Phenolic Extract of Fennel and Reinforced by Magnesium Oxide: In Vivo - in Vitro

Authors: Mazouzi Nourdjihane, K. Boutemak, A. Haddad, Y. Chegreouche

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In the last decades, biodegradable polymers have been considered as one of the most popular options for the delivery of drugs and various conventional doses. The film forming system (FFS) can be used in topical, transdermal, ophthalmic, oral and gastric applications. Recently this system has focused on improving drug delivery, which can promote drug release. In this context, the aim of this study is to create polymeric film-forming systems for the stomach and to evaluate and test their gastroprotective effects, comparing the effects of changes in composition on film characteristics. It uses a plant-derived polyphenol extract extracted from fennel to demonstrate anti-inflammatory activity in the film. The films are made from hydroxypropyl methylcellulose polymer and different types of plastic, glycerol and polyethylene glycol. The ffs properties show that MgO-glycerol-reinforced hydroxypropylmethylcellulose (HPMC-MgO-Gly) is better than that based on MgO-PEG-reinforced hydroxypropylmethylcellulose (HPMC-MgO-PEG). It is durable, has a faster drying time and allows for maximum recovery. Water vapor strength and blowing speed and other additions show another advantage of HPMC-MgO-Gly compared to HPMC-MgO-PEG, indicating good adhesion between the support (top) and film production. In this study, the gastroprotective effect of fennel phenol extract was found, showing that this plant material has a gastroprotective effect on ulcers and that the film can absorb the active substance.

Keywords: film formin system, hydroxypropyl methylcellulose, magnesium oxide, in vivo

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157 Amine Hardeners with Carbon Nanotubes Dispersing Ability for Epoxy Coating Systems

Authors: Szymon Kugler, Krzysztof Kowalczyk, Tadeusz Spychaj

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An addition of carbon nanotubes (CNT) can simultaneously improve many features of epoxy coatings, i.e. electrical, mechanical, functional and thermal. Unfortunately, this nanofiller negatively affects visual properties of the coatings, such as transparency and gloss. The main reason for the low visual performance of CNT-modified epoxy coatings is the lack of compatibility between CNT and popular amine curing agents, although epoxy resins based on bisphenol A are indisputable good CNT dispersants. This is a serious obstacle in utilization of the coatings in advanced applications, demanding both high transparency and electrical conductivity. The aim of performed investigations was to find amine curing agents exhibiting affinity for CNT, and ensuring good performance of epoxy coatings with them. Commercially available CNT was dispersed in epoxy resin, as well as in different aliphatic, cycloaliphatic and aromatic amines, using one of two dispergation methods: ultrasonic or mechanical. The CNT dispersions were subsequently used in the preparation of epoxy coating compositions and coatings on a transparent substrate. It was found that amine derivative of bio-based cardanol, as well as modified o-tolylbiguanide exhibit significant CNT, dispersing properties, resulting in improved transparent/electroconductive performance of epoxy coatings. In one of prepared coating systems just 0.025 wt.% (250 ppm) of CNT was enough to obtain coatings with semi conductive properties, 83% of transparency as well as perfect chemical resistance to methyl-ethyl ketone and improved thermal stability. Additionally, a theory of the influence of amine chemical structure on CNT dispersing properties was proposed.

Keywords: bio-based cardanol, carbon nanotubes, epoxy coatings, tolylbiguanide

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156 Characterization of Volatile Compounds in Meat Lamb Fed in Different Algeria Pasture

Authors: Nabila Berrighi, Kaddour Bouderoua, Maria Khossif, Gema Nieto, Gaspar Ros

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Ruminant meat is an important source of nutrients and is also of high sensory value. However, the importance and nature of these characteristics depend on ruminant nutrition. The objective of this study is to assess the effect of two Algerian feeding systems applied in the steppic rearing area of Djelfa and in the highlands one of Tiaret on the growth performance of lambs and on their meat quality, especially on their aroma compounds of meat. At the beginning of the experiment, lambs had an average body weight of 34.04 kg, and 35.40 kg for the group reared at Highland (0% concentrate) and Steppe (30% concentrate), respectively. The incorporation of the concentrated feed in Steppe had a significant effect on slaughter weight compared to lambs fed only on pasture (Highland) (49.72 Kg vs. 42.06 Kg, P<0.05). Beyond the first month, animals from the Steppe one showed better weight gains compared to those from Highland (14.32Kg vs. 8.02 Kg, respectively, P<0,05). After slaughter, samples from the Longissimus thoracis were removed and analyzed. The results point to significant differences in the amounts of many of the predominant volatile compounds between both groups (p<0.05), such as Hexanal, 2-methyl-3-furanthiol and nonanal (8.92 μg/kg vs. 4.57 μg/kg), (8.88 μg/kg vs. 7.45 μg/kg) and (2.09 μ/kg vs. 1.02 μg/kg) associated with smells of green, boiling meat and orange fruit, respectively. These compounds, measured by olfactometry, derived from the oxidation of lipids and appear to be responsible for the characteristic flavor of lamb meat in the steppe compared to that generated by meat from animals from the Highland pastures. The Algerian Steppe ecosystem is very interesting for outdoor sheep breeding, which allows to obtain attractive sensory quality and in the production of typical lamb meat that can be considered as a label.

Keywords: falvour, growth performance, lamb meat, steppe pasture

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155 Identification and Characterization of Inhibitors of Epoxide Hydrolase from Trichoderma reesei

Authors: Gabriel S. De Oliveira, Patricia P. Adriani, Christophe Moriseau, Bruce D. Hammock, Felipe S. Chambergo

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Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have high biotechnological interest for the drug design and chemistry transformation for industries. In this study, we describe the identification of substrates and inhibitors of epoxide hydrolase enzyme from the filamentous fungus Trichoderma reesei (TrEH), and these inhibitors showed the fungal growth inhibitory activity. We have used the cloned enzyme and expressed in E. coli to develop the screening in the library of fluorescent substrates with the objective of finding the best substrate to be used in the identification of good inhibitors for the enzyme TrEH. The substrate (3-phenyloxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester showed the highest specific activity and was chosen for the next steps of the study. The inhibitors screening was performed in the library with more than three thousand molecules and we could identify the 6 best inhibitors. The IC50 of these molecules were determined in nM and all the best inhibitors have urea or amide in their structure, because It has been recognized that these groups fit well in the hydrolase catalytic pocket of the epoxide hydrolases. Then the growth of T. reesei in PDA medium containing these TrEH inhibitors was tested, and fungal growth inhibition activity was demonstrated with more than 60% of inhibition of fungus growth in the assay with the TrEH inhibitor with the lowest IC50. Understanding how this EH enzyme from T. reesei responds to inhibitors may contribute for the study of fungal metabolism and drug design against pathogenic fungi.

Keywords: epoxide hydrolases, fungal growth inhibition, inhibitor, Trichoderma reesei

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154 The Effects of Extraction Methods on Fat Content and Fatty Acid Profiles of Marine Fish Species

Authors: Yesim Özogul, Fethiye Takadaş, Mustafa Durmus, Yılmaz Ucar, Ali Rıza Köşker, Gulsun Özyurt, Fatih Özogul

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It has been well documented that polyunsaturated fatty acids (PUFAs), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have beneficial effects on health, regarding prevention of cardiovascular diseases, cancer and autoimmune disorders, development the brain and retina and treatment of major depressive disorder etc. Thus, an adequate intake of omega PUFA is essential and generally marine fish are the richest sources of PUFA in human diet. Thus, this study was conducted to evaluate the efficiency of different extraction methods (Bligh and Dyer, soxhlet, microwave and ultrasonics) on the fat content and fatty acid profiles of marine fish species (Mullus babatus, Upeneus moluccensis, Mullus surmuletus, Anguilla anguilla, Pagellus erythrinus and Saurida undosquamis). Fish species were caught by trawl in Mediterranean Sea and immediately iced. After that, fish were transported to laboratory in ice and stored at -18oC in a freezer until the day of analyses. After extracting lipid from fish by different methods, lipid samples were converted to their constituent fatty acid methyl esters. The fatty acid composition was analysed by a GC Clarus 500 with an autosampler (Perkin Elmer, Shelton, CT, USA) equipped with a flame ionization detector and a fused silica capillary SGE column (30 m x 0.32 mm ID x 0.25 mm BP20 0.25 UM, USA). The results showed that there were significant differences (P < 0.05) in fatty acids of all species and also extraction methods affected fat contents and fatty acid profiles of fish species.

Keywords: extraction methods, fatty acids, marine fish, PUFA

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153 Molecular Engineering of High-Performance Nanofiltration Membranes from Intrinsically Microporous Poly (Ether-Ether-Ketone)

Authors: Mahmoud A. Abdulhamid

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Poly(ether-ether-ketone) (PEEK) has received increased attention due to its outstanding performance in different membrane applications including gas and liquid separation. However, it suffers from a semi-crystalline morphology, bad solubility and low porosity. To fabricate membranes from PEEK, the usage of harsh acid such as sulfuric acid is essential, regardless its hazardous properties. In this work, we report the molecular design of poly(ether-ether-ketones) (iPEEKs) with intrinsic porosity character, by incorporating kinked units into PEEK backbone such as spirobisindane, Tröger's base, and triptycene. The porous polymers were used to fabricate stable membranes for organic solvent nanofiltration application. To better understand the mechanism, we conducted molecular dynamics simulations to evaluate the possible interactions between the polymers and the solvents. Notable enhancement in separation performance was observed confirming the importance of molecular engineering of high-performance polymers. The iPEEKs demonstrated good solubility in polar aprotic solvents, a high surface area of 205–250 m² g⁻¹, and excellent thermal stability. Mechanically flexible nanofiltration membranes were prepared from N-methyl-2-pyrrolidone dope solution at iPEEK concentrations of 19–35 wt%. The molecular weight cutoff of the membranes was fine-tuned in the range of 450–845 g mol⁻¹ displaying 2–6 fold higher permeance (3.57–11.09 L m⁻² h⁻¹ bar⁻¹) than previous reports. The long-term stabilities were demonstrated by a 7 day continuous cross-flow filtration.

Keywords: molecular engineering, polymer synthesis, membrane fabrication, liquid separation

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152 Halogenated Methoxy- and Methyl-benzoic Acids: Joint Experimental and DFT Study For Molecular Structure, Vibrational Analysis, and Other Molecular Properties

Authors: Boda Sreenivas, Lyathakula Ravindranath, Kanugula Srishailam, Byru Venkatram Reddy

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Extensive research into the optimized structure and molecular properties of 3-Flouro-2-methylbenzoicacid(FMB), 3-Chloro-2-methoxybenzoicacid (CMB), and 3-Bromo-2-methylbenzoicacid (BMB) was carried out using FT-IR, FT-Raman and UV-Visible spectra, as well as theoretically using the DFT approach with B3LYPfunctional in conjunction with 6-311++G(d,p) basis set. The optimized structure was determined by evaluating torsional scans about free rotation bonds. Structure parameters, harmonic vibrational frequencies, potential energy distribution(PED), and infrared and Raman intensities were computed. The computational results from the DFT approach, such asFT-IR, FT-Raman, and UV-Visible spectra, were compared with the experimental results and found good agreement. Observed and calculated frequencies agreed with an rms error of 8.42, 6.60, and 6.95 cm-1 for FMB, CMB, and BMB, respectively. Unambiguous vibrational assignments were made for all fundamentals using PED and eigenvectors. The electronic HOMO-LUMO, H-bonding, and strong conjugative interactions across different molecular entities are discussed using experimental and simulated Ultraviolet-Visible spectra. The title molecules' molecular properties such as dipole moment, mean polarizability, and first-order hyperpolarizability, were calculated to study their non-linear optical (NLO) behavior. The chemical reactivity descriptors and mapped electrostatic surface potential (MESP) were also evaluated. Natural bond orbital (NBO) analysis was used to examine the stability of molecules resulting from hyperconjugative interactions and charge delocalization.

Keywords: ftir/raman spectra, DFT, NLO, homo-lumo, NBO, halogenated benzoic acids

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151 Fatty Acids and Inflammatory Protein Biomarkers in Freshly Frozen Plasma Samples from Patients with and without COVID-19

Authors: Alaa Hamed Habib

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The Coronavirus disease 2019 (COVID-19) is a viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and associated with systemic inflammation. Inflammation is an important process that follows infection and facilitates the repair of damaged tissue. Polyunsaturated fatty acids play an important role in the inflammatory process. These lipids can target transcription factors to modulate gene expression and protein function. Here, we evaluated whether differences in basal levels of different types of biomarkers can be detected in freshly frozen plasma samples from patients with and without COVID19. Fatty acid methyl ester (FAME) analysis showed a decrease in arachidic acid and myristic acid, but an increase in caprylic acid, palmitic acid, and eicosenoic acid in the plasma of COVID-19 patients compared to non-COVID19 patients. Multiple chemokines, including IP-10, MCP-1, and MIP-1 beta, were increased in the COVID-19 group compared to the non-COVID-19 group. Similarly, cytokines including IL-1 alpha and IL-8, and cell adhesion and inflammatory response markers including ICAM-1 and E-selectin were greater in the plasma of COVID-19 patients compared to non-COVID-19 patients. A baseline signature of specific polyunsaturated fatty acids, cytokines, and chemokines present in the plasma after COVID-19 viral infection may serve as biomarkers that can be useful in various applications, including determination of the severity of infection, an indication of disease prognosis and consideration for therapeutic options.

Keywords: MARKS, COVID 19, UEVS NON COVIDS, kidneys, nanoparticles

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150 Quantification of Hydrogen Sulfide and Methyl Mercaptan in Air Samples from a Waste Management Facilities

Authors: R. F. Vieira, S. A. Figueiredo, O. M. Freitas, V. F. Domingues, C. Delerue-Matos

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The presence of sulphur compounds like hydrogen sulphide and mercaptans is one of the reasons for waste-water treatment and waste management being associated with odour emissions. In this context having a quantifying method for these compounds helps in the optimization of treatment with the goal of their elimination, namely biofiltration processes. The aim of this study was the development of a method for quantification of odorous gases in waste treatment plants air samples. A method based on head space solid phase microextraction (HS-SPME) coupled with gas chromatography - flame photometric detector (GC-FPD) was used to analyse H2S and Metil Mercaptan (MM). The extraction was carried out with a 75-μm Carboxen-polydimethylsiloxane fiber coating at 22 ºC for 20 min, and analysed by a GC 2010 Plus A from Shimadzu with a sulphur filter detector: splitless mode (0.3 min), the column temperature program was from 60 ºC, increased by 15 ºC/min to 100 ºC (2 min). The injector temperature was held at 250 ºC, and the detector at 260 ºC. For calibration curve a gas diluter equipment (digital Hovagas G2 - Multi Component Gas Mixer) was used to do the standards. This unit had two input connections, one for a stream of the dilute gas and another for a stream of nitrogen and an output connected to a glass bulb. A 40 ppm H2S and a 50 ppm MM cylinders were used. The equipment was programmed to the selected concentration, and it automatically carried out the dilution to the glass bulb. The mixture was left flowing through the glass bulb for 5 min and then the extremities were closed. This method allowed the calibration between 1-20 ppm for H2S and 0.02-0.1 ppm and 1-3.5 ppm for MM. Several quantifications of air samples from inlet and outlet of a biofilter operating in a waste management facility in the north of Portugal allowed the evaluation the biofilters performance.

Keywords: biofiltration, hydrogen sulphide, mercaptans, quantification

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149 Protective Potential of Hyperhalophilic Diatoms Extract Against Lead Induced Oxidative Stress in Rats and Human HepG2 and HEK293 Cells Line

Authors: Wassim Guermazi, Saoussan Boukhris, Neila Annabi Trabelsi, Tarek Rebai, Alya Sellami-Kamoun, Habib Ayadi

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This work investigates the protective effects of the microalga Halamphora sp. extract (H. Ext) as a natural product on lead-intoxicated liver and kidney human cells in vitro and in vivo on rats wistar. HepG2 cells line derived from human hepatocellular carcinoma and HEK293 cells line derived from human embryonic kidney were used for the in vitro study. The analysis of the fatty acids methyl esters of the extract was performed by a GC/MS. Four groups of rats, each of which was composed of six animals, were used for the in vivo experiment. The pretreatment of HepG2 and HEK293 cells line with the extract (100 µg mL-1) significantly (p < 0.05) protected against cytotoxicity induced by lead exposure. In vivo, the biochemical parameters in serum, namely malondialdehyde level (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities, were measured in supernatants of organ homogenates. H. Ext was found to be rich in fatty acids, essentially palmitic and palmitoleic accounting respectively 29.46% and 42.07% of total fatty acids. Both in vitro and in vivo, the co-treatment with H. Ext allowed the protection of the liver and kidney cells structure, as well as the significant preservation of normal antioxidant and biochemical parameters in rats. Halamphora extract rich in fatty acids has been proven to be effective in protection against Pb-induced toxicity.

Keywords: microalga extract, human cells line, fatty acid, lead exposure, oxidative stress, rats

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148 Anti-Infective Potential of Selected Philippine Medicinal Plant Extracts against Multidrug-Resistant Bacteria

Authors: Demetrio L. Valle Jr., Juliana Janet M. Puzon, Windell L. Rivera

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From the various medicinal plants available in the Philippines, crude ethanol extracts of twelve (12) Philippine medicinal plants, namely: Senna alata L. Roxb. (akapulko), Psidium guajava L. (bayabas), Piper betle L. (ikmo), Vitex negundo L. (lagundi), Mitrephora lanotan (Blanco) Merr. (Lanotan), Zingiber officinale Roscoe (luya), Curcuma longa L. (Luyang dilaw), Tinospora rumphii Boerl (Makabuhay), Moringga oleifera Lam. (malunggay), Phyllanthus niruri L. (sampa-sampalukan), Centella asiatica (L.) Urban (takip kuhol), and Carmona retusa (Vahl) Masam (tsaang gubat) were studied. In vitro methods of evaluation against selected Gram-positive and Gram-negative multidrug-resistant (MDR), bacteria were performed on the plant extracts. Although five of the plants showed varying antagonistic activities against the test organisms, only Piper betle L. exhibited significant activities against both Gram-negative and Gram-positive multidrug-resistant bacteria, exhibiting wide zones of growth inhibition in the disk diffusion assay, and with the lowest concentrations of the extract required to inhibit the growth of the bacteria, as supported by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Further antibacterial studies of the Piper betle L. leaf, obtained by three extraction methods (ethanol, methanol, supercritical CO2), revealed similar inhibitory activities against a multitude of Gram-positive and Gram-negative MDR bacteria. Thin layer chromatography (TLC) assay of the leaf extract revealed a maximum of eight compounds with Rf values of 0.92, 0.86, 0.76, 0.53, 0.40, 0.25, 0.13, and 0.013, best visualized when inspected under UV-366 nm. TLC- agar overlay bioautography of the isolated compounds showed the compounds with Rf values of 0.86 and 0.13 having inhibitory activities against Gram-positive MDR bacteria (MRSA and VRE). The compound with an Rf value of 0.86 also possesses inhibitory activity against Gram-negative MDR bacteria (CRE Klebsiella pneumoniae and MBL Acinetobacter baumannii). Gas Chromatography-Mass Spectrometry (GC-MS) was able to identify six volatile compounds, four of which are new compounds that have not been mentioned in the medical literature. The chemical compounds isolated include 4-(2-propenyl)phenol and eugenol; and the new four compounds were ethyl diazoacetate, tris(trifluoromethyl)phosphine, heptafluorobutyrate, and 3-fluoro-2-propynenitrite. Phytochemical screening and investigation of its antioxidant, cytotoxic, possible hemolytic activities, and mechanisms of antibacterial activity were also done. The results showed that the local variant of Piper betle leaf extract possesses significant antioxidant, anti-cancer and antimicrobial properties, attributed to the presence of bioactive compounds, particularly of flavonoids (condensed tannin, leucoanthocyanin, gamma benzopyrone), anthraquinones, steroids/triterpenes and 2-deoxysugars. Piper betle L. is also traditionally known to enhance wound healing, which could be primarily due to its antioxidant, anti-inflammatory and antimicrobial activities. In vivo studies on mice using 2.5% and 5% of the ethanol leaf extract cream formulations in the excised wound models significantly increased the process of wound healing in the mice subjects, the results and values of which are at par with the current antibacterial cream (Mupirocin). From the results of the series of studies, we have definitely proven the value of Piper betle L. as a source of bioactive compounds that could be developed into therapeutic agents against MDR bacteria.

Keywords: Philippine herbal medicine, multidrug-resistant bacteria, Piper betle, TLC-bioautography

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