Search results for: spontaneous activity
5637 A High-Throughput Enzyme Screening Method Using Broadband Coherent Anti-stokes Raman Spectroscopy
Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai
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Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy
Procedia PDF Downloads 1345636 Study of the Antimicrobial Potential Of a Rich Polyphenolic Extract Obtained from Cytisus scoparius
Authors: Lorena G. Calvo, Marta Lores, Trinidad de Miguel
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Natural extracts containing high polyphenolic concentration possess antibacterial and antifungal activity. The present research characterizes a hydro-organic extract with a high polyphenolic content as an antimicrobial candidate. As a result of this composition, the extract showed pronounced bioactivities with potential uses in agricultural, veterinary, pharmaceutical, and cosmetic industries. Polyphenol compounds were extracted by using hydro-organic solvent mixtures from the shrub Cytisus scoparius. The in vitro antimicrobial activity of this extract was evaluated on Gram-positive and Gram-negative bacteria and the fungus Candida albicans. Microbial species investigated, Bacillus cereus, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, are causing agents of several human and animal diseases. The extract showed activity against all tested species. So, it could be used for the development of biocides to control a wide range of pathogenic agents and contribute to the creation of economic and eco-friendly alternatives to antibiotics.Keywords: antimicrobial properties, antioxidant properties, Cytisus scoparius, polyphenolic extract
Procedia PDF Downloads 1385635 Contextualization and Localization: Acceptability of the Developed Activity Sheets in Science 5 Integrating Climate Change Adaptation
Authors: Kim Alvin De Lara
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The research aimed to assess the level of acceptability of the developed activity sheets in Science 5 integrating climate change adaptation of grade 5 science teachers in the District of Pililla school year 2016-2017. In this research, participants were able to recognize and understand the importance of environmental education in improving basic education and integrating them in lessons through localization and contextualization. The researcher conducted the study to develop a material to use by Science teachers in Grade 5. It served also as a self-learning resource for students. The respondents of the study were the thirteen Grade 5 teachers teaching Science 5 in the District of Pililla. Respondents were selected purposively and identified by the researcher. A descriptive method of research was utilized in the research. The main instrument was a checklist which includes items on the objectives, content, tasks, contextualization and localization of the developed activity sheets. The researcher developed a 2-week lesson in Science 5 for 4th Quarter based on the curriculum guide with integration of climate change adaptation. The findings revealed that majority of respondents are female, 31 years old and above, 10 years above in teaching science and have units in master’s degree. With regards to the level of acceptability, the study revealed developed activity sheets in science 5 is very much acceptable. In view of the findings, lessons in science 5 must be contextualized and localized to improve to make the curriculum responds, conforms, reflects, and be flexible to the needs of the learners, especially the 21st century learners who need to be holistically and skillfully developed. As revealed by the findings, it is more acceptable to localized and contextualized the learning materials for pupils. Policy formation and re-organization of the lessons and competencies in Science must be reviewed and re-evaluated. Lessons in science must also be integrated with climate change adaptation since nowadays, people are experiencing change in climate due to global warming and other factors. Through developed activity sheets, researcher strongly supports environmental education and believes this to serve as a way to instill environmental literacy to students.Keywords: activity sheets, climate change adaptation, contextualization, localization
Procedia PDF Downloads 3185634 Isolation, Characterization and Screening of Antimicrobial Producing Actinomycetes from Sediments of Persian Gulf
Authors: H. Alijani, M. Jabari, S. Matroodi, H. Zolqarnein, A. Sharafi, I. Zamani
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Actinomycetes, Gram-positive bacteria, are interesting as a main producer of secondary metabolites and are important industrially and pharmaceutically. The marine environment is a potential source for new actinomycetes, which can provide novel bioactive compounds and industrially important enzymes. The aims of this study were to isolate and identify novel actinomycetes from Persian Gulf sediments and screen these isolates for the production of secondary metabolites, especially antibiotics, Using phylogenetic (16S rRNA gene sequence), morphological and biochemical analyses. 15 different actinomycete strains from Persian Gulf sediments at a depth of 5-10 m were identified. DNA extraction was done using Cinnapure DNA Kit. PCR amplification of 16S rDNA gene was performed using F27 and R1492 primers. Phylogenetic tree analysis was performed using the MEGA 6 software. Most of the isolated strains belong to the genus namely Streptomyces (14), followed by Nocardiopsis (1). Antibacterial assay of the isolates supernatant was performed using a standard disc diffusion assay with replication (n=3). The results of disk diffusion assay showed that most active strain against Proteus volgaris and Bacillus cereus was AMJ1 (16.46±0.2mm and 13.78±0.2mm, respectively), against Salmonella sp. AMJ7 was the most effective strain (10.13±0.2mm), and AMJ1 and AHA5 showed more inhibitory activity against Escherichia coli (8.04±0.02 mm and 8.2±0.03 ). The AMJ6 strain showed best antibacterial activity against Klebsiella sp. (8.03±0.02mm). Antifungal activity of AMJ2 showed that it was most active strain against complex (16.05±0.02mm) and against Aspergillus flavus strain AMJ1 was most active strain (16.4±0.2mm) and highest antifungal activity against Trichophyton mentagrophytes, Microsporum gyp serum and Candida albicans, were shown by AHA1 (21.03±0.02mm), AHA3 and AHA7 (18±0.03mm), AMJ6 (21.03±0.2mm) respectively. Our results revealed that the marine actinomycetes of Persian Gulf sediments were potent source of novel antibiotics and bioactive compounds and indicated that the antimicrobial metabolites were extracellular. Most of the secondary metabolites and antibiotics are extracellular in nature and extracellular products of actinomycetes show potent antimicrobial activities.Keywords: antibacterial activity, antifungal activity, marine actinomycetes, Persian Gulf
Procedia PDF Downloads 2935633 Search of Сompounds with Antimicrobial and Antifungal Activity in the Series of 1-(2-(1H-Tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas
Authors: O. Antypenko, I. Vasilieva, S. Kovalenko
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Investigations for new effective and less toxic antimicrobials agents are always up-to-date. The tetrazole derivatives are quite interesting objects as for synthesis as well as for pharmacological screening. Thus, some derivatives of tetrazole demonstrated antimicrobial activity, namely 5-phenyl-tetrazolo[1,5-c]quinazoline was effective one against Staphylococcus aureus and Esherichia faecalis (MIC = 250 mg/L). Besides, investigation of the 9-bromo(chloro)-5-morpholin(piperidine)-4-yl-tetrazolo[1,5-c]quinazoline’s antimicrobial activity against Esherichia coli and Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus revealed that sensitivity of Gram-positive bacteria to the compounds was higher than that of Gram-negative bacteria. So, our previously synthesized, 31 derivatives of 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas were decided to test for their in vitro antibacterial activity against Gram-positive bacteria (Staphylococcus aureus ATCC 25923, Enterobacter aerogenes, Enterococcus faecalis ATCC 29212), Gram-negative bacteria (Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 25922, Klebsiella pneumoniae 68) and antifungal properties against Candida albicans ATCC 885653. Agar-diffusion method was used for determination of the preliminary activity compared to well-known reference antimicrobials. All the compounds were dissolved in DMSO at a concentration of 100 μg/disk, using inhibition zone diameter (IZD, mm) as a measure for the antimicrobial activity. The most active turned to be 3 structures, that inhibited several bacterial strains: 1-ethyl-3-(5-fluoro-2-(1H-tetrazol-5-yl)phenyl)urea (1), 1-(4-bromo-2-(1H-tetrazol-5-yl)-phenyl)-3-(4-(trifluoromethyl)phenyl)urea (2) and 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (3). IZM (mm) was 40 (Escherichia coli), 25 (Klebsiella pneumonia) for compound 1; 12 (Pseudomonas aeruginosa), 15 (Staphylococcus aureus), 10 (Enterococcus faecalis) for compound 2; 25 (Staphylococcus aureus), 15 (Enterococcus faecalis) for compound 3. The most sensitive to the activity of the substances were Gram-negative bacteria Pseudomonas aeruginosa. While none of compound effected on Candida albicans. Speaking about, reference drugs: Amikacin (30 µg/disk) showed 27 and Ceftazide (30 µg/disk) 25 against Pseudomonas aeruginosa. That is, unfortunately, higher than studied 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas. Obtained results will be used for further purposeful optimization of the leading compounds in the more effective antimicrobials because of the ever-mounting problem of microorganism’s resistance.Keywords: antimicrobial, antifungal, compounds, 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas
Procedia PDF Downloads 3545632 Impact of Ethnomedicinal Plants on Toothpaste Improvement
Authors: Muna Jalal Ali, Essam A. Makky, Mashitah M. Yusoff
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Objectives: The aim of this study to evaluate the antimicrobial susceptibility of combined toothpaste with medicinal plants and the relations between the commercial toothpaste to its price and the patient age as well. Materials and Methods: Oral isolates of different patients aged 3 to 60 years were obtained, purified, and tested against four different ethnomedicinal plant extracts for antimicrobial activity. A total of 10 different commercial toothpastes (different brands and prices) were collected from the market, and the combined action of the medicinal plants and toothpaste was studied. Results: We found a higher bacterial population in the age group of 3–40 years than the group of 40–60 years, with approximately 44% and 32%, respectively. The combined action of ethanolic extract (alone) against oral isolates showed a synergistic effect, with 32.20, 30.50, and 25.42% for combinations A (Ci/Ca), B (Ci/Ca/P), and C (Ci/Ca/P/N), respectively. By contrast, the combined action of ethnomedicinal plants with 10 different toothpastes improved the antimicrobial sensitivity by 60, 100, and 0% for combinations A, B, and C respectively. Clinical relevance: The ethanolic extract of only combinations A and B with commercial toothpaste showed high antibacterial activity against oral isolates and the effectiveness of toothpaste is not related to the price.Keywords: microbial evolution, oral isolates, ethnomedicinal plants, antimicrobial activity, toothpaste
Procedia PDF Downloads 3015631 A Fast Method for Graphene-Supported Pd-Co Nanostructures as Catalyst toward Ethanol Oxidation in Alkaline Media
Authors: Amir Shafiee Kisomi, Mehrdad Mofidi
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Nowadays, fuel cells as a promising alternative for power source have been widely studied owing to their security, high energy density, low operation temperatures, renewable capability and low environmental pollutant emission. The nanoparticles of core-shell type could be widely described in a combination of a shell (outer layer material) and a core (inner material), and their characteristics are greatly conditional on dimensions and composition of the core and shell. In addition, the change in the constituting materials or the ratio of core to the shell can create their special noble characteristics. In this study, a fast technique for the fabrication of a Pd-Co/G/GCE modified electrode is offered. Thermal decomposition reaction of cobalt (II) formate salt over the surface of graphene/glassy carbon electrode (G/GCE) is utilized for the synthesis of Co nanoparticles. The nanoparticles of Pd-Co decorated on the graphene are created based on the following method: (1) Thermal decomposition reaction of cobalt (II) formate salt and (2) the galvanic replacement process Co by Pd2+. The physical and electrochemical performances of the as-prepared Pd-Co/G electrocatalyst are studied by Field Emission Scanning Electron Microscopy (FESEM), Energy Dispersive X-ray Spectroscopy (EDS), Cyclic Voltammetry (CV), and Chronoamperometry (CHA). Galvanic replacement method is utilized as a facile and spontaneous approach for growth of Pd nanostructures. The Pd-Co/G is used as an anode catalyst for ethanol oxidation in alkaline media. The Pd-Co/G not only delivered much higher current density (262.3 mAcm-2) compared to the Pd/C (32.1 mAcm-2) catalyst, but also demonstrated a negative shift of the onset oxidation potential (-0.480 vs -0.460 mV) in the forward sweep. Moreover, the novel Pd-Co/G electrocatalyst represents large electrochemically active surface area (ECSA), lower apparent activation energy (Ea), higher levels of durability and poisoning tolerance compared to the Pd/C catalyst. The paper demonstrates that the catalytic activity and stability of Pd-Co/G electrocatalyst are higher than those of the Pd/C electrocatalyst toward ethanol oxidation in alkaline media.Keywords: thermal decomposition, nanostructures, galvanic replacement, electrocatalyst, ethanol oxidation, alkaline media
Procedia PDF Downloads 1455630 Job Satisfaction among Public and Private Universities in Egypt Related to Organizational and Personal Aspects
Authors: Reem Alkadeem
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This study aims at evaluating the overall satisfaction of faculty members and relating it to organizational and personal aspects in Egyptian public and private universities. These aspects are identified through an extensive study of all factors that might affect job satisfaction. The most influencing parameters selected are academics’ demographics, human resource management, organizational profile, workload, teamwork skills, recognition, autonomy, teaching activity, research activity, and motivation. A questionnaire of 94 questions was used to assess job satisfaction and the previously mentioned parameters. It was distributed among seven hundred members of different universities in Egypt. Two hundred and twenty-seven faculty members responded. This sample was gathered from twelve universities and The Supreme Council of Universities. The ANOVA showed a significant relationship (p < 0.05) between eight of the selected parameters and job satisfaction. These parameters are age, rank, human resource management, profile of organizational characteristics, workload, recognition, teaching activity, and motivation.Keywords: job satisfaction, higher education, organizational profile, Egyptian universities
Procedia PDF Downloads 4755629 Antimicrobial Activity of Nauclea lotifolia (African Peach) Crude Extracts against Some Pathogenic Microorganism
Authors: Muhammad Isah Legbo
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The phytochemical screening and antimicrobial activity of Nauclea lotifolia fruit, leaf and stem-bark extracts at various concentration of (20.0,10.0, 5.0, and 2.5 mg/ml) were evaluated against some pathogenic microorganisms such as Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Staphylococcus aureus, Aspergillus niger and Candida albicans. The antimicrobial activity was assayed using agar well diffusion method. The result obtained show appreciable inhibitory effort of acetone, aqueous and methanolic extracts of Nauclea lotifolia. However, result obtained was less active compared to that of the control antibiotic (Ciprofloxacillin). The minimum inhibitory concentration (MIC) was determined using serial doubling dilution method and ranged from 5.0-10.0mg/ml, while the minimum bactericidal concentration (MBC) was determined by plating various dilution of extracts without turbidity and the result ranged from 5.0-7.5mg/ml. The phytochemical screening revealed the presence of alkaloid, anthraquinones, flavonoids, resin, steroid and saponin. The activities of the plant extract therefore justify their utilization in the treatment of various ailments associated with the test organism.Keywords: Nauclea, lotifolia, antimicrobial, pathogens, saponin, extract
Procedia PDF Downloads 4065628 An Overview of Structure Based Activity Outcomes of Pyran Derivatives Against Alzheimer’s Disease
Authors: Faisal Almalki
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Pyran is a heterocyclic group containing oxygen that possesses a variety of pharmacological effects. Pyran is also one of the most prevalent structural subunits in natural products, such as xanthones, coumarins, flavonoids, benzopyrans, etc. Additionally demonstrating the neuroprotective properties of pyrans is the fact that this heterocycle has recently attracted the attention of scientists worldwide. Alzheimer's Disease (AD) treatment and diagnosis are two of the most critical research objectives worldwide. Increased amounts of extracellular senile plaques, intracellular neurofibrillary tangles, and a progressive shutdown of cholinergic basal forebrain neuron transmission are often related with cognitive impairment. This review highlights the various pyran scaffolds of natural and synthetic origin that are effective in the treatment of AD. For better understanding synthetic compounds are categorized as different types of pyran derivatives like chromene, flavone, xanthone, xanthene, etc. The discussion encompasses both the structure-activity correlations of these compounds as well as their activity against AD. Because of the intriguing actions that were uncovered by these pyran-based scaffolds, there is no question that they are at the forefront of the search for potential medication candidates that could treat Alzheimer's disease.Keywords: alzheimer’s disease, pyran, coumarin, xanthone
Procedia PDF Downloads 645627 Antimicrobial Activity of a Single Wap Domain (SWD)-Containing Protein from Litopenaeus vannamei against Vibrio parahaemolyticus Acute Hepatopancreatic Necrosis Disease (AHPND)
Authors: Suchao Donpudsa, Suwattana Visetnan, Anchalee Tassanakajon, Vichien Rimphanitchayakit
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The Single Wap Domain (SWD) is a type III crustin antimicrobial peptide whose function is to defense the host animal against the bacterial infection by means of antimicrobial and antiproteinase activities. A study of LvSWD from Litopenaeus vannamei is reported herein about its activities and function against bacteria, particularly the Vibrio parahaemolyticus AHPND (VPAHPND) that causes acute hepatopancreatic necrosis disease. The over-expressed mature recombinant (r)LvSWD exhibits antimicrobial activity against both Gram-positive and Gram-negative bacteria, especially VPAHPND. With four times the MIC of rLvSWD, the treated post larval shrimp infected by VPAHPND is able to survive longer with the 50% survival rate as long as 78 h as compared to 36 h of the infected shrimp without rLvSWD. To a certain extent, we have demonstrated that the rLvSWD can be applied to protect the post larval shrimp.Keywords: crustin, Litopenaeus vannamei, Vibrio parahaemolyticus AHPND, antimicrobial activity
Procedia PDF Downloads 2085626 Developing Students’ Academic Writing Skills through Scientific Reading: Using Questions and Answer Activities
Authors: Makhim Artikova, Shavkat Duschanov
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So far, there have been a plethora of attempts to improve learners’ academic writing skills. However, this issue remains to be a real concern among the majority of students, especially those who are standing on their academic life threshold. The purpose of this research is improving students’ academic writing skills through 'Questions and Answer Reading' activities. Using well-prepared and well-chosen reading materials (from textbooks, scientific journals, or magazines) and applying questions and answer activities in the classroom facilitate learners to become great critical readers. Furthermore, it boosts their writing skills, which are the most crucial part of students’ personal and academic developments. In this activity, the class is divided into small groups of four. Then, the instructor will give students whether one section of the text or full text asking them to read and to find unfamiliar words within the group. After discovering the meaning of unknown words, each group has to share their findings with the class. In the next stage of the activity, students should be asked to create questions in a group based on the given reading material. Follow by each group should ask the other groups their questions which are an excellent opportunity to challenge leads to improve critical thinking skills. In the last part, the students are asked to write the text or article summary, which is the activity core that pilots to the writing skills perfection. This engaging activity highlights the effectiveness of incorporating reading materials into the classroom when it comes to improving students’ composition writings. Structural writing after every reading activity resulted in improving students’ coherence and cohesion in writing well-organized essays. Having experimented with high school 9th and 11th-grade students, implementing reading activities into the classroom is proved to be a productive tool to enhance one’s academic writing skills. In the future, this method planning to be implemented among university students.Keywords: academic writing, coherence and cohesion, questions and answer activities, scientific reading
Procedia PDF Downloads 1075625 Extracellular Enzymes as Promising Soil Health Indicators: Assessing Response to Different Land Uses Using Long-Term Experiments
Authors: Munisath Khandoker, Stephan Haefele, Andy Gregory
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Extracellular enzymes play a key role in soil organic carbon (SOC) decomposition and nutrient cycling and are known indicators for soil health; however, it is not understood how these enzymes respond to different land uses and their relationships to other soil properties have not been extensively reviewed. The relationships among the activities of three soil enzymes: β-glucosaminidase (NAG), phosphomonoesterase (PHO) and β-glucosidase (GLU), were examined. The impact of soil organic amendments, soil types and land management on soil enzyme activities were reviewed, and it was hypothesized that soils with increased SOC have increased enzyme activity. Long-term experiments at Rothamsted Research Woburn and Harpenden sites in the UK were used to evaluate how different management practices affect enzyme activity involved in carbon (C) and nitrogen (N) cycling in the soil. Samples were collected from soils with different organic treatments such as straw, farmyard manure (FYM), compost additions, cover crops and permanent grass cover to assess whether SOC can be linked with increased levels of enzymatic activity and what influence, if any, enzymatic activity has on total C and N in the soil. Investigating the interactions of important enzymes with soil characteristics and SOC can help to better understand the health of soils. Studies on long-term experiments with known histories and large datasets can better help with this. SOC tends to decrease during land use changes from natural ecosystems to agricultural systems; therefore, it is imperative that agricultural lands find ways to increase and/or maintain SOC in the soil.Keywords: biological soil health indicators, extracellular enzymes, soil health, soil, microbiology
Procedia PDF Downloads 665624 Quantitative Structure Activity Relationship and Insilco Docking of Substituted 1,3,4-Oxadiazole Derivatives as Potential Glucosamine-6-Phosphate Synthase Inhibitors
Authors: Suman Bala, Sunil Kamboj, Vipin Saini
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Quantitative Structure Activity Relationship (QSAR) analysis has been developed to relate antifungal activity of novel substituted 1,3,4-oxadiazole against Candida albicans and Aspergillus niger using computer assisted multiple regression analysis. The study has shown the better relationship between antifungal activities with respect to various descriptors established by multiple regression analysis. The analysis has shown statistically significant correlation with R2 values 0.932 and 0.782 against Candida albicans and Aspergillus niger respectively. These derivatives were further subjected to molecular docking studies to investigate the interactions between the target compounds and amino acid residues present in the active site of glucosamine-6-phosphate synthase. All the synthesized compounds have better docking score as compared to standard fluconazole. Our results could be used for the further design as well as development of optimal and potential antifungal agents.Keywords: 1, 3, 4-oxadiazole, QSAR, multiple linear regression, docking, glucosamine-6-phosphate synthase
Procedia PDF Downloads 3315623 Canthin-6-One Alkaloid Inhibits NF-κB and AP-1 Activity: An Inhibitory Action At Transcriptional Level
Authors: Fadia Gafri, Kathryn Mckintosh, Louise Young, Alan Harvey, Simon Mackay, Andrew Paul, Robin Plevin
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Nuclear factor-kappa B (NF-κB) is a ubiquitous transcription factor found originally to play a key role in regulating inflammation. However considerable evidence links this pathway to the suppression of apoptosis, cellular transformation, proliferation and invasion (Aggarwal et al., 2006). Moreover, recent studies have also linked inflammation to cancer progression making NF-κB overall a promising therapeutic target for drug discovery (Dobrovolskaia & Kozlov, 2005). In this study we examined the effect of the natural product canthin-6-one (SU182) as part of a CRUK small molecule drug discovery programme for effects upon the NF-κB pathway. Initial studies demonstrated that SU182 was found to have good potency against the inhibitory kappa B kinases (IKKs) at 30M in vitro. However, at concentrations up to 30M, SU182 had no effect upon TNFα stimulated loss in cellular IκBα or p65 phosphorylation in the keratinocyte cell line NCTC2544. Nevertheless, 30M SU182 reduced TNF-α / PMA-induced NF-κB-linked luciferase reporter activity to (22.9 ± 5%) and (34.6± 3 %, P<0.001) respectively, suggesting an action downstream of IKK signalling. Indeed, SU182 neither decreased NF-κB-DNA binding as assayed by EMSA nor prevented the translocation of p65 (NF-κB) to the nucleus assessed by immunofluorescence and subcellular fractionation. In addition to the inhibition of transcriptional activity of TNFα-induced NF-κB reporter activity SU182 significantly reduced PMA-induced AP-1-linked luciferase reporter activity to about (48± 9% at 30M, P<0.001) . This mode of inhibition was not sufficient to prevent the activation of NF-κB dependent induction of other proteins such as COX-2 and iNOS, or activated MAP kinases (p38, JNK and ERK1/2) in LPS stimulated RAW 264.7 macrophages. Taken together these data indicate the potential for SU182 to interfere with the transcription factors NF-κB and AP-1 at transcriptional level. However, no potential anti-inflammatory effect was indicated, further investigation for other NF-κB dependent proteins linked to survival are also required to identify the exact mechanism of action.Keywords: Canthin-6-one, NF-κB, AP-1, phosphorylation, Nuclear translocation, DNA-binding activity, inflammatory proteins.
Procedia PDF Downloads 4515622 A Report of 5-Months-Old Baby with Balanced Chromosomal Rearrangements along with Phenotypic Abnormalities
Authors: Mohit Kumar, Beklashwar Salona, Shiv Murti, Mukesh Singh
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We report here a case of five-months old male baby, born as second child of non-consanguineous parents with no considerable history of genetic abnormality which was referred to our cytogenetic laboratory for chromosomal analysis. Physical dysmorphic facial features including mongoloid face, cleft palate, simian crease, and developmental delay were observed. We present this case with unique balanced autosomal translocation of t(3;10)(p21;p13). The risk of phenotypic abnormalities based on de novo balanced translocation was estimated to be 7%. The association of balanced chromosomal rearrangement with Down syndrome features such as multiple congenital anomalies, facial dysmorphism and congenital heart anomalies are very rare in a 5-months old male child. Trisomy-21 is not uncommon in chromosomal abnormality with the birth defect and balanced translocations are frequently observed in patients with secondary infertility or recurrent spontaneous abortion (RSA). Two ml heparinized peripheral blood cells cultured in RPMI-1640 for 72 hours supplemented with 20% fetal bovine serum, phytohemagglutinin (PHA), and antibiotics were used for chromosomal analysis. A total 30 metaphases images were captured using Olympus-BX51 microscope and analyzed using Bio-view karyotyping software through GTG-banding (G bands by trypsin and Giemsa) according to International System for Human Cytogenetic Nomenclature 2016. The results showed balanced translocation between short arm of chromosome # 3 and short arm of chromosome # 10. The karyotype of the child was found to be 46,XY,t(3;10)(p21; p13). Chromosomal abnormalities are one of the major causes of birth defect in new born babies. Also, balanced translocations are frequently observed in patients with secondary infertility or recurrent spontaneous abortion. The index case presented with dysmorphic facial features and had a balanced translocation 46,XY,t(3;10)(p21;p13). This translocation with break points at (p21; p13) has not been reported in the literature in a child with facial dysmorphism. To the best of our knowledge, this is the first report of novel balanced translocation t(3;10) with break points in a child with dysmorphic features. We found balanced chromosomal translocation instead of any trisomy or unbalanced aberrations along with some phenotypic abnormalities. Therefore, we suggest that such novel balanced translocation with abnormal phenotype should be reported in order to enable the pathologist, pediatrician, and gynecologist to have a better insight into the intricacies of chromosomal abnormalities and their associated phenotypic features. We hypothesized that dysmorphic features as seen in this case may be the result of change in the pattern of genes located at the breakpoint area in balanced translocations or may be due to deletion or mutation of genes located on the p-arm of chromosome # 3 and p-arm of chromosome # 10.Keywords: balanced translocation, karyotyping, phenotypic abnormalities, facial dimorphisms
Procedia PDF Downloads 2025621 Factors Influencing Walking in Bandar Baru Bangi, Malaysia
Authors: Zeinab Aliyas
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Walking is known as the most common type of physical activity that helps mental and physical health of people. In the recent years, promoting walking activity in neighborhood areas and cities become as one of the important issues in terms of sustainable cities. Therefore the study aimed to investigate the influence of fear of crime and personal barriers as social and personal factor respectively on neighborhood walking. 464 questionnaires in Bandar Baru Bangi in Malaysia was distributed to collect data, and finally, 424 questionnaires were qualified to be used in the study. The Smart-PLS was used to analyze the data. The findings of the study revealed that individual barriers and fear of crime both have significant influence on the level of walking behavior in the neighborhood area. It was found that fear of crime has higher influence on walking behavior in comparison to individual factors. The finding of this study can help urban researcher and planner to know the significant influence of crime safety and individual attitudes on the level of walking activity.Keywords: fear of crime, neighborhood walking, personal barriers, residential neighborhood
Procedia PDF Downloads 1745620 Optimising Participation in Physical Activity Research for Adults with Intellectual Disabilities
Authors: Yetunde M. Dairo, Johnny Collett, Helen Dawes
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Background and Aim: Engagement with physical activity (PA) research is poor among adults with intellectual disabilities (ID), particularly in those from residential homes. This study explored why, by asking managers of residential homes, adults with ID and their carers. Methods: Participants: A convenient sample of 23 individuals from two UK local authorities, including a group of ID residential home managers, adults with ID and their support staff. Procedures: A) Residential home managers (n=6) were asked questions about their willingness to allow their residents to participate in PA research; B) eleven adults with ID and their support workers (n=6) were asked questions about their willingness to accept 7-day accelerometer monitoring and/or the International Physical Activity Questionnaire-short version (IPAQ-s) as PA measures. The IPAQ-s was administered by the researcher and they were each provided with samples of accelerometers to try on. Results: A) Five out of six managers said that the burden of wearing the accelerometer for seven days would be too high for the people they support, the majority of whom might be unable to express their wishes. They also said they would be unwilling to act as proxy respondents for the same reason. Additionally, they cited time pressure, understaffing, and reluctance to spend time on the research paperwork as further reasons for non-participation. B) All 11 individuals with ID completed the IPAQ-s while only three accepted the accelerometer, one of whom was deemed inappropriate to wear it. Reasons for rejecting accelerometers included statements from participants of: ‘too expensive’, ‘too heavy’, ‘uncomfortable’, and two people said they would not want to wear it for more than one day. All adults with ID (11) and their support workers (6) provided information about their physical activity levels through the IPAQ-s. Conclusions: Care home managers are a barrier to research participation. However, adults with ID would be happy for the IPAQ-s as a PA measure, but less so for the 7-day accelerometer monitoring. In order to improve participation in this population, the choice of PA measure is considered important. Moreover, there is a need for studies exploring how best to engage ID residential home managers in PA research.Keywords: intellectual disability, physical activity measurement, research engagement, research participation
Procedia PDF Downloads 3005619 Production of Human BMP-7 with Recombinant E. coli and B. subtilis
Authors: Jong Il Rhee
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The polypeptide representing the mature part of human BMP-7 was cloned and efficiently expressed in Escherichia coli and Bacillus subtilis, which had a clear band for hBMP-7, a homodimeric protein with an apparent molecular weight of 15.4 kDa. Recombinant E.coli produced 111 pg hBMP-7/mg of protein hBMP-7 through IPTG induction. Recombinant B. subtilis also produced 350 pg hBMP-7/ml of culture medium. The hBMP-7 was purified in 2 steps using an FPLC system with an ion exchange column and a gel filtration column. The hBMP-7 produced in this work also stimulated the alkaline phosphatase (ALP) activity in a dose-dependent manner, i.e. 2.5- and 8.9-fold at 100 and 300 ng hBMP-7/ml, respectively, and showed intact biological activity.Keywords: B. subtilis, E. coli, fermentation, hBMP-7
Procedia PDF Downloads 4265618 Effect of Fat Percentage and Prebiotic Composition on Proteolysis, ACE-Inhibitory and Antioxidant Activity of Probiotic Yogurt
Authors: Mohammad B. HabibiNajafi, Saeideh Sadat Fatemizadeh, Maryam Tavakoli
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In recent years, the consumption of functional foods, including foods containing probiotic bacteria, has come to notice. Milk proteins have been identified as a source of angiotensin-I-converting enzyme )ACE( inhibitory peptides and are currently the best-known class of bioactive peptides. In this study, the effects of adding prebiotic ingredients (inulin and wheat fiber) and fat percentage (0%, 2% and 3.5%) in yogurt containing probiotic Lactobacillus casei on physicochemical properties, degree of proteolysis, antioxidant and ACE-inhibitory activity within 21 days of storage at 5 ± 1 °C were evaluated. The results of statistical analysis showed that the application of prebiotic compounds led to a significant increase in water holding capacity, proteolysis and ACE-inhibitory of samples. The degree of proteolysis in yogurt increases as storage time elapses (P < 0.05) but when proteolysis exceeds a certain threshold, this trend begins to decline. Also, during storage time, water holding capacity reduced initially but increased thereafter. Moreover, based on our findings, the survival of Lactobacillus casei in samples treated with inulin and wheat fiber increased significantly in comparison to the control sample (P < 0.05) whereas the effect of fat percentage on the survival of probiotic bacteria was not significant (P = 0.095). Furthermore, the effect of prebiotic ingredients and the presence of probiotic cultures on the antioxidant activity of samples was significant (P < 0.05).Keywords: probiotic yogurt, proteolysis, ACE-inhibitory, antioxidant activity
Procedia PDF Downloads 2465617 ZnO / TiO2 Nanoparticles for Degradation of Cyanide Ion
Authors: Masoumeh Tabatabaee, Zahra Shahryarzadeh, Masoud R. Shishebor
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Advanced oxidation process (AOPs) is alternative method for the complete degradation many organic pollutants. When a photocatalyst absorbs radiation whose energy hν > Eg an ē from its filled valance band (VB) is promoted to its conduction band (CB) and valance band holes h+ are formed. Electron would reduce any available species, including O2, water and hydroxide ion to form hydroxyl radicals. ZnO and TiO2 are important photocatalysts with high catalytic activity that have attracted much research attention. TiO2 can only absorb a small portion of solar spectrum in the UV region and many methods such as dye sensitization, doping of other metals and using TiO2 with another semiconductor have been used to improve the photocatalytic activity of TiO2 under solar irradiation. Studies have shown that the use of metal oxides or sulfide such as WO3, MoO3, SiO2, MgO, ZnO, and CdS with TiO2 can significantly enhance the photocatalytic activity of TiO2. Due to similarity of photodegradation mechanism of ZnO with TiO2, it is a suitable semiconductor using with TiO2 and recently nanosized bicomponent TiO2-ZnO photocatalysts were prepared and used for degradation of some pollutants. In this study, Nano-sized ZnO/TiO2 composite was synthesized. Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscope (SEM) were used to characterize the structure and morphology of it. The effect of photocatalytic activity of prepared ZnO/TiO2 on the degradation of cyanide ion under UV was investigated. The effect of various parameters such as ZnO/TiO2 concentration, amount of photocatalyst, amount of H2O2, initial dye or cyanide ion concentration, pH and irradiation time on were investigated. Results show that more than 95% of 4 mgL-1 cyanide ion degraded after 60-min reaction time and under UV irradiation.Keywords: photodegradation, ZnO/TiO2, nanoparticle, cyanide ion
Procedia PDF Downloads 3885616 Development of Composite Materials for CO2 Reduction and Organic Compound Decomposition
Authors: H. F. Shi, C. L. Zhang
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Visible-light-responsive g-C3N4/NaNbO3 nanowires photocatalysts were fabricated by introducing polymeric g-C3N4 on NaNbO3 nanowires. The microscopic mechanisms of interface interaction, charge transfer and separation, as well as the influence on the photocatalytic activity of g-C3N4/NaNbO3 composite were systematic investigated. The HR-TEM revealed that an intimate interface between C3N4 and NaNbO3 nanowires formed in the g-C3N4/NaNbO3 heterojunctions. The photocatalytic performance of photocatalysts was evaluated for CO2 reduction under visible-light illumination. Significantly, the activity of g-C3N4/NaNbO3 composite photocatalyst for photoreduction of CO2 was higher than that of either single-phase g-C3N4 or NaNbO3. Such a remarkable enhancement of photocatalytic activity was mainly ascribed to the improved separation and transfer of photogenerated electron-hole pairs at the intimate interface of g-C3N4/NaNbO3 heterojunctions, which originated from the well-aligned overlapping band structures of C3N4 and NaNbO3. Pt loaded NaNbO3-xNx (Pt-NNON), a visible-light-sensitive photocatalyst, was synthesized by an in situ photodeposition method from H2PtCl6•6H2O onto NaNbO3-xNx (NNON) sample. Pt-NNON exhibited a much higher photocatalytic activity for gaseous 2-propanol (IPA) degradation under visible-light irradiation in contrast to NNON. The apparent quantum efficiency (AQE) of Pt-NNON sample for IPA photodegradation achieved up to 8.6% at the wavelength of 419 nm. The notably enhanced photocatalytic performance was attributed to the promoted charge separation and transfer capability in the Pt-NNON system. This work suggests that surface nanosteps possibly play an important role as an electron transfer at high way, which facilitates to the charge carrier collection onto Pt rich zones and thus suppresses recombination between photogenerated electrons and holes. This method can thus be considered as an excellent strategy to enhance photocatalytic activity of organic decomposition in addition to the commonly applied noble metal doping method.Keywords: CO2 reduction, NaNbO3, nanowires, g-C3N4
Procedia PDF Downloads 1985615 Nitrogen Fixation in Hare Gastrointestinal Tract
Authors: Tatiana A. Kuznetsova, Maxim V. Vechersky, Natalia V. Kostina, Marat M. Umarov, Elena I. Naumova
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One of the main problems of nutrition of phytophagous animals is the insufficiency of protein in their forage. Usually, symbiotic microorganisms highly contribute both to carbohydrates and nitrogen compounds of the food. But it is not easy to utilize microbial biomass in the large intestine and caecum for the animals with hindgut fermentation. So that, some animals, as well hares, developed special mechanism of contribution of such biomass - obligate autocoprophagy, or reingestion. Hares have two types of feces - the hard and the soft. Hard feces are excreted at night, while hares are vigilance ("foraging period"), and the soft ones (caecotrophs) are produced and reingested in the day-time during hares "resting-period". We examine the role of microbial digestion in providing nitrogen nutrition of hare (Lepus europaeus). We determine the ability of nitrogen fixation in fornix and stomach body, small intestine, caecum and colon of hares' gastro-intestinal tract in two main period of hares activity - "resting-period" (day time) and "foraging period" (late-evening and very-early-morning). We use gas chromatography to measure levels of nitrogen fixing activity (acetylene reduction). Nitrogen fixing activity was detected in the contents of all analyzed parts of the gastrointestinal tract. Maximum values were recorded in the large intestine. Also daily dynamics of the process was detected. Thus, during hare “resting-period” (caecotrophs formation) N2-fixing activity was significantly higher than during “foraging period”, reaching 0,3 nmol C2H4/g*h. N2-fixing activity in the gastrointestinal tract can allocate to significant contribution of nitrogen fixers to microbial digestion in hare and confirms the importance of coprophagy as a nitrogen source in lagomorphs.Keywords: coprophagy, gastrointestinal tract, lagomorphs, nitrogen fixation
Procedia PDF Downloads 3555614 Human Lens Metabolome: A Combined LC-MS and NMR Study
Authors: Vadim V. Yanshole, Lyudmila V. Yanshole, Alexey S. Kiryutin, Timofey D. Verkhovod, Yuri P. Tsentalovich
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Cataract, or clouding of the eye lens, is the leading cause of vision impairment in the world. The lens tissue have very specific structure: It does not have vascular system, the lens proteins – crystallins – do not turnover throughout lifespan. The protection of lens proteins is provided by the metabolites which diffuse inside the lens from the aqueous humor or synthesized in the lens epithelial layer. Therefore, the study of changes in the metabolite composition of a cataractous lens as compared to a normal lens may elucidate the possible mechanisms of the cataract formation. Quantitative metabolomic profiles of normal and cataractous human lenses were obtained with the combined use of high-frequency nuclear magnetic resonance (NMR) and ion-pairing high-performance liquid chromatography with high-resolution mass-spectrometric detection (LC-MS) methods. The quantitative content of more than fifty metabolites has been determined in this work for normal aged and cataractous human lenses. The most abundant metabolites in the normal lens are myo-inositol, lactate, creatine, glutathione, glutamate, and glucose. For the majority of metabolites, their levels in the lens cortex and nucleus are similar, with the few exceptions including antioxidants and UV filters: The concentrations of glutathione, ascorbate and NAD in the lens nucleus decrease as compared to the cortex, while the levels of the secondary UV filters formed from primary UV filters in redox processes increase. That confirms that the lens core is metabolically inert, and the metabolic activity in the lens nucleus is mostly restricted by protection from the oxidative stress caused by UV irradiation, UV filter spontaneous decomposition, or other factors. It was found that the metabolomic composition of normal and age-matched cataractous human lenses differ significantly. The content of the most important metabolites – antioxidants, UV filters, and osmolytes – in the cataractous nucleus is at least ten fold lower than in the normal nucleus. One may suppose that the majority of these metabolites are synthesized in the lens epithelial layer, and that age-related cataractogenesis might originate from the dysfunction of the lens epithelial cells. Comprehensive quantitative metabolic profiles of the human eye lens have been acquired for the first time. The obtained data can be used for the analysis of changes in the lens chemical composition occurring with age and with the cataract development.Keywords: cataract, lens, NMR, LC-MS, metabolome
Procedia PDF Downloads 3105613 To What Extent Does Physical Activity and Standard of Competition Affect Quantitative Ultrasound (QUS) Measurements of Bone in Accordance with Muscular Strength and Anthropometrics in British Young Males?
Authors: Joseph Shanks, Matthew Taylor, Foong Kiew Ooi, Chee Keong Chen
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Introduction: Evidences of relationship between bone, muscle and standard of competition among young British population is limited in literature. The current literature recognises the independent and synergistic effects of fat free and fat mass as the stimulus for osteogenesis. This study assessed the extent to which physical activity (PA) and standard of competition (CS) influences quantitative ultrasound (QUS) measurements of bone on a cross-sectional basis accounting for muscular strength and anthropometrics in British young males. Methods: Pre-screening grouped 66 males aged 18-25 years into controls (n=33) and district level athletes (DLAs) (n=33) as well as low (n=21), moderate (n=23) and high (n=22) physical activity categories (PACs). All participants underwent QUS measurements of bone (4 sites, i.e. dominant distal radius (DR), dominant mid-shaft tibia (DT), non-dominant distal radius (NR) and non-dominant mid-shaft tibia (NT)), isokinetic strength tests (dominant and non-dominant knee flexion and extension) and anthropometric measurements. Results: There were no significant differences between any of the groups with respect to QUS measurements of bone at all sites with regards to PACs or CS. Significant higher isokinetic strength values were observed in DLAs than controls (p < 0.05), and higher than low PACs (p < 0.05) at 60o.s-1 of concentric and eccentric measurements. No differences in subcutaneous fat thickness were found between all the groups (CS or PACs). Percentages of body fat were significantly higher (p < .05) in low than high PACs and CS groups. There were significant positive relationships between non dominant radial speed of sound and fat free mass at both DR (r=0.383, p=0.001) and NR (r=0.319, p=0.009) sites in all participants. Conclusion: The present study findings indicated that muscular strength and body fat are closely related to physical activity level and standard of competition. However, bone health status reflected by quantitative ultrasound (QUS) measurements of bone is not related to physical activity level and standard of competition in British young males.Keywords: bone, muscular strength, physical activity, standard of competition
Procedia PDF Downloads 5115612 Measurement of Radon Exhalation Rate, Natural Radioactivity, and Radiation Hazard Assessment in Soil Samples from the Surrounding Area of Kasimpur Thermal Power Plant Kasimpur (U. P.), India
Authors: Anil Sharma, Ajay Kumar Mahur, R. G. Sonkawade, A. C. Sharma, R. Prasad
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In coal fired thermal power stations, large amount of fly ash is produced after burning of coal. Fly ash is spread and distributed in the surrounding area by air and may be deposited on the soil of the region surrounding the power plant. Coal contains increased levels of these radionuclides and fly ash may increase the radioactivity in the soil around the power plant. Radon atoms entering into the pore space from the mineral grain are transported by diffusion and advection through this space until they in turn decay or are released into the atmosphere. In the present study, Soil samples were collected from the region around a Kasimpur Thermal Power Plant, Kasimpur, Aligarh (U.P.). Radon activity, radon surface exhalation and mass exhalation rates were measured using “sealed can technique” using LR 115-type II nuclear track detectors. Radon activities vary from 92.9 to 556.8 Bq m-3 with mean value of 279.8 Bq m-3. Surface exhalation rates (EX) in these samples are found to vary from 33.4 to 200.2 mBq m-2 h-1 with an average value of 100.5 mBq m-2 h-1 whereas, Mass exhalation rates (EM) vary from 1.2 to 7.7 mBq kg-1 h-1 with an average value of 3.8 mBq kg-1 h-1. Activity concentrations of radionuclides were measured in these samples by using a low level NaI (Tl) based gamma ray spectrometer. Activity concentrations of 226Ra 232Th and 40K vary from 12 to 49 Bq kg-1, 24 to 49 Bq kg-1 and 135 to 546 Bq kg-1 with overall mean values of 30.3 Bq kg-1, 38.5 Bq kg-1 and 317.8 Bq kg-1, respectively. Radium equivalent activity has been found to vary from 80.0 to 143.7 Bq kg-1 with an average value of 109.7 Bq kg-1. Absorbed dose rate varies from 36.1 to 66.4 nGy h-1 with an average value of 50.4 nGy h-1 and corresponding outdoor annual effective dose varies from 0.044 to 0.081 mSv with an average value of 0.061 mSv. Values of external and internal hazard index Hex, Hin in this study vary from 0.21 to 0.38 and 0.27 to 0.50 with an average value of 0.29 and 0.37, Respectively. The results will be discussed in light of various factors.Keywords: natural radioactivity, radium equivalent activity, absorbed dose rate, gamma ray spectroscopy
Procedia PDF Downloads 3555611 Human Xanthine Oxidase Inhibitory Effect, in vivo Antioxidant Activity of Globularia alypum L. Extracts
Authors: N. Boussoualim, H. Trabsa, I. Krache, S. Aouachria, S. Boumerfeg, L. Arrar, A. Baghiani
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The aim of this study consisted in evaluating the antioxidant in vivo properties, anti-hemolytic and XOR inhibitory effect of Globularia alypum L. (GA) extracts. GA was submitted to extraction and fractionation to give crude (CrE), chloroformique (ChE), ethyle acetate (EAE) and aqueos (AqE) extracts. Total polyphenols contents of GA extracts were determined; EAE is the most rich in polyphenols (157,74±5,27 mg GAE/mg of extract). GA Extracts inhibited XO in a concentration-dependent manner, the EAE showed the highest inhibitory properties on the XOR activity (IC50=0,083±0,001 mg/ml), followed by CrE and ChE. The antioxidant activities of the CrE, EAE, and AqE were tested by an in vivo assay in mice, the plasma ability to inhibit DPPH radical was measured, The CrE was found to exhibit the greatest scavenger activity with 48.41±2.763%, followed by AqE and EAE (40.54±7.51% and 41.79±1.654%, respectively). Total antioxidant capacity of red blood cells was measured, from the kinetics of hemolysis obtained. The calculated HT50 reveal an extension of time for half hemolysis in all treated groups compared with the control group. CrE increase significantly HT50 (112,8±2,427). The hemolysis is lagged, indicating that endogenous antioxidants in the erythrocytes can trap radicals to protect them against free-radical-induced hemolysis. Antimicrobial activities of the extracts were determined by the disc diffusion method. Test microorganisms were; 4 Gram positive, 7 gram negative bacteria, most active extracts were EAE and CrE. We deduce a great relationship between the effect on the extracts antibacterial effect and their contents in flavonoid.Keywords: Globularia alypum, Xanthine oxidoreductase, in vivo-antioxidant activity, hemolysis, polyphenol
Procedia PDF Downloads 3255610 The Effect of Bisphenol A and Its Selected Analogues on Antioxidant Enzymes Activity in Human Erythrocytes
Authors: Aneta Maćczak, Bożena Bukowska, Jaromir Michałowicz
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Bisphenols are one of the most widely used chemical compounds worldwide. They are used in the manufacturing of polycarbonates, epoxy resins and thermal paper which are applied in plastic containers, bottles, cans, newspapers, receipt and other products. Among these compounds, bisphenol A (BPA) is produced in the highest amounts. There are concerns about endocrine impact of BPA and its other toxic effects including hepatotoxicity, neurotoxicity and carcinogenicity on human organism. Moreover, BPA is supposed to increase the incidence the obesity, diabetes and heart disease. For this reason the use of BPA in the production of plastic infant feeding bottles and some other consumers products has been restricted in the European Union and the United States. Nowadays, BPA analogues like bisphenol F (BPF) and bisphenol S (BPS) have been developed as alternative compounds. The replacement of BPA with other bisphenols contributed to the increase of the exposure of human population to these substances. Toxicological studies have mainly focused on BPA. In opposite, a small number of studies concerning toxic effects of BPA analogues have been realized, which makes impossible to state whether those substituents are safe for human health. Up to now, the mechanism of bisphenols action on the erythrocytes has not been elucidated. That is why, the aim of this study was to assess the effect of BPA and its selected analogues such as BPF and BPS on the activity of antioxidant enzymes, i.e. catalase (EC 1.11.1.6.), glutathione peroxidase (E.C.1.11.1.9) and superoxide dismutase (EC.1.15.1.1) in human erythrocytes. Red blood cells in respect to their function (transport of oxygen) and very well developed enzymatic and non-enzymatic antioxidative system, are useful cellular model to assess changes in redox balance. Erythrocytes were incubated with BPA, BPF and BPS in the concentration ranging from 0.5 to 100 µg/ml for 24 h. The activity of catalase was determined by the method of Aebi (1984). The activity of glutathione peroxidase was measured according to the method described by Rice-Evans et al. (1991), while the activity of superoxide dismutase (EC.1.15.1.1) was determined by the method of Misra and Fridovich (1972). The results showed that BPA and BPF caused changes in the antioxidative enzymes activities. BPA decreased the activity of examined enzymes in the concentration of 100 µg/ml. We also noted that BPF decreased the activity of catalase (5-100 µg/ml), glutathione peroxidase (50-100 µg/ml) and superoxide dismutase (25-100 µg/ml), while BPS did not cause statistically significant changes in investigated parameters. The obtained results suggest that BPA and BPF disrupt redox balance in human erythrocytes but the observed changes may occur in human organism only during occupational or subacute exposure to these substances.Keywords: antioxidant enzymes, bisphenol A, bisphenol a analogues, human erythrocytes
Procedia PDF Downloads 4655609 Identification and Characterization of Antimicrobial Peptides Isolated from Entophytic Bacteria and Their Activity against Multidrug-Resistance Gram-Negative Bacteria in South Korea
Authors: Maryam Beiranvand
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Multi-drug resistance in various microorganisms has increased globally in many healthcare facilities. Less effective antimicrobial activity of drug therapies for infection control becomes trouble. Since 1980, no new type of antimicrobial drug has been identified, even though combinations of antibiotic drugs have been discovered almost every decade. Between 1981 and 2006, over 70% of novel pharmaceuticals and chemical agents came from natural sources. Microorganisms have yielded almost 22,000 natural compounds. The identification of antimicrobial components from endophytes bacteria could help overcome the threat posed by multi-drug resistant strains. The project aims to analyze and identify antimicrobial peptides isolated from entophytic bacteria and their activity against multidrug-resistant Gram-negative bacteria in South Korea. Endophytic Paenibacillus polymyxa. 4G3 isolated from the plant, Gynura procumbery exhibited considerable antimicrobial activity against Methicillin-resistant Staphylococcus aureus, and Escherichia coli. The Rapid Annotations using Subsystems Technology showed that the total size of the draft genome was 5,739,603bp, containing 5178 genes with 45.8% G+C content. Genome annotation using antiSMASH version 6.0.0 was performed, which predicted the most common types of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS). In this study, diethyl aminoethyl cellulose (DEAEC) resin was used as the first step in purifying for unknown peptides, and then the target protein was identified using hydrophilic and hydrophobic solutions, optimal pH, and step-by-step tests for antimicrobial activity. This crude was subjected to C18 chromatography and elution with 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% methanol, respectively. Only the fraction eluted with 20% -60% methanol demonstrated good antimicrobial activity against MDR E. coli. The concentration of the active fragment was measured by the Brad-ford test, and Protein A280 - Thermo Fisher Scientific at the end by examining the SDS PAGE Resolving Gel, 10% Acrylamide and purity were confirmed. Our study showed that, based on the combined results of the analysis and purification. P polymyxa. 4G3 has a high potential exists for producing novel functions of polymyxin E and bacitracin against bacterial pathogens.Keywords: endophytic bacteria, antimicrobial activity, antimicrobial peptide, whole genome sequencing analysis, multi -drug resistance gram negative bacteria
Procedia PDF Downloads 665608 Production and Characterisation of Lipase from a Novel Streptomyces.sp - Its Molecular Identification
Authors: C. Asha Poorna, N. S. Pradeep
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The biological function of lipase is to catalyze the hydrolysis of triacylglycerols to give free fatty acid, diacylglycerols, mono-acylglycerols and glycerol. They constitute the most important group of biocatalysts for biotechnological applications. The aim of the present study was to identify the lipolytic activity of Streptomyces sp. From soil sample collected from the sacred groves of southern Kerala. The culture conditions of the isolate were optimised and the enzyme was purified and characterised. The purification was attempted with acetone precipitation. The isolate observed to have high lipolytic activity and identified to be of Streptomyces strain. The purification was attempted with acetone precipitation. The purified enzyme observed to have an apparent molecular mass of ~60kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme showed maximum activity at 60oC and pH-8. The lipase showed tolerance towards different organic solvents like ethanol and methanol that are commonly used in transesterification reactions to displace alcohol from triglycerides contained in renewable resources to yield fatty acid alkyl esters known as biodiesel.Keywords: lipase, Streptomyces, biodiesel, fatty acid, transesterification
Procedia PDF Downloads 321