Search results for: biological sludge
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2548

Search results for: biological sludge

1288 Biological Expressions of Hamilton’s Rule in Human Populations: The Deep Psychological Influence of Defensive and Offensive Motivations Found in Human Conflicts and Sporting Events

Authors: Monty Vacura

Abstract:

Hamilton’s Rule is a universal law of biology expressed in protists, plants and animals. When applied to human populations, this model explains: 1) Origin of religion in society as a biopsychological need naturally selected to increase population size; 2) Instincts of racism expressed through intergroup competition; 3) Simultaneous selection for human cooperation and conflict, love and hate; 4) Places Dawkins’s selfish gene as the r, relationship variable; 5) Flipping the equation variable themes (close relationship to distant relationship, and benefit to threat) the new equation can now be used to identify the offensive and defensive sides of conflict; 6) Connection between sporting events and instinctive social messaging for stimulating offensive and defensive responses; 6) Pathway to reduce human sacrifice through manipulation of variables. This paper discusses the deep psychological influences of Hamilton’s Rule. Suggestions are provided to reduce human deaths via our instinctive sacrificial behavior, by consciously monitoring Hamilton’s Rule variables highlighted throughout our media outlets.

Keywords: psychology, Hamilton’s rule, evolution, human instincts

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1287 Study of the Behavior of Copper Immersed in Sea Water of the Bay of Large Agadir by Electrochemical Methods

Authors: Aicha Chaouay, Lahsen Bazzi, Mustapha Hilali

Abstract:

Seawater has chemical and biological characteristics making it particularly aggressive in relation to the corrosion of many materials including copper and steels low or moderate allies. Note that these materials are widely used in the manufacture of port infrastructure in the marine environment. These structures are exposed to two types of corrosion including: general corrosion and localized corrosion caused by the presence of sulfite-reducing micro-organisms. This work contributes to the study of the problematic related to bacterial contamination of the marine environment of large Agadir and evaluating the impact of this pollution on the corrosion resistance of copper. For the realization of this work, we conducted monthly periodic draws between (October 2012 February 2013) of seawater from the Anza area of the Bay of Agadir. Thus, after each sampling, a study of the electro chemical corrosion behavior of copper was carried out. Electro chemical corrosion parameters such as the corrosion potential, the corrosion current density, the charge transfer resistance and the double layer capacity were evaluated. The electro chemical techniques used in this work are: the route potentiodynamic polarization curves and electro chemical impedance.

Keywords: Bay of Agadir, microbial contamination, seawater (Morocco), corrosion, copper

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1286 Effects of Post-Emergence Herbicides on Soil Micro-Flora and Nitrogen Fixing Bacteria in Pea Field

Authors: Ali M. Zaid, Muftah Mayouf, Yahya Said Farouj

Abstract:

The effect of post emergence herbicides on soil micro-flora and nitrogen fixing bacteria was studied in pea field. Pea (Pisum sativum) was grown and treated with one or a mixture of two of several herbicides 2 weeks after sowing. Soil samples were collected 2 weeks after herbicides application. Average number of colony forming units per gram of soil of bacteria, actinomycetes and fungi were determined. Average number of nodules per plant was obtained at the end of the growing season. The results of the study showed MCPB, Bentazon, MCPB+Fluozifop-p-butyl, Bentazon+Fluozifop-p-butyl, Metribuzin, Flouzifop-p-butyl+Metribuzin, Cycloxydin, and Sethoxydin increased the population of soil fungi, with 4 to 10 times compared with the control. The herbicides used showed no significant effects on nitrogen fixing bacteria. The effects of herbicides on soil bacteria and actinomycetes were different. The study showed the use of herbicides could influence the biological balance of soil microflora, which has an important role in soil fertility and microbial ecosystem.

Keywords: herbicides, post emergence, nitrogen fixing bacteria, environmental systems

Procedia PDF Downloads 379
1285 Discovering Traditional Plants Used by Indigenous People in the Tropical Rainforest of Malaysia for the Treatment of Malaria

Authors: Izdihar Ismail, Alona C. Linatoc, Maryati Mohamed

Abstract:

The tropical rainforest of Malaysia is known for its rich biological diversity and high endemicity. The potential for these forests to hold the cure for many diseases and illnesses is high and much is yet to be discovered. This study explores the richness of the tropical rainforest of Endau-Rompin National Park in Johor, Malaysia in search of plants traditionally used by the indigenous people in the treatment of malaria and malaria-like symptoms. Seven species of plants were evaluated and tested for antiplasmodial activities. Different plant parts were subjected to methanolic and aqueous extractions. A total of 24 extracts were evaluated by histidine-rich protein II (HRP2) assay against K1 strain of Plasmodium falciparum chloroquine-resistant. Ten extracts showed significant inhibition of the growth of P. falciparum. Phytochemical screening of the same extracts revealed the presence of alkaloids, flavonoids, terpenoids and anthraquinones. This study affirms that tropical rainforests may still hold undiscovered cures for many diseases and illnesses that have inflicted millions of people worldwide. The species studied herein have not known to have been studied elsewhere before.

Keywords: Endau-Rompin, malaria, Malaysia, tropical rainforest, traditional knowledge

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1284 Phytochemical and Biological Study of Chrozophora oblongifolia

Authors: Al-Braa Kashegari, Ali M. El-Halawany, Akram A. Shalabi, Sabrin R. M. Ibrahim, Hossam M. Abdallah

Abstract:

Chemical investigation of Chrozophora oblongifolia resulted in the isolation of five major compounds that were identified as apeginin-7-O-glucoside (1), quercetin-3-O-glucuronic acid (2), quercetin-3-O-glacturonic acid (3), rutin (4), and 1,3,6-trigalloyl glucose (5). The identity of isolated compounds was assessed by different spectroscopic methods, including one- and two-dimensional NMR. The isolated compounds were tested for their antioxidant activity using different assays viz., DPPH, FRAP, ABTS, ORAC, and metal chelation effects. In addition, the inhibition of target enzymes involved in the metabolic syndrome, such as alpha-glucosidase and pancreatic lipase, were carried out. Moreover, the effect of the compounds on the advanced glycation end-products (AGEs) as one of the major complications of oxidative stress and hyperglycemia in metabolic syndromes were carried out using BSA‐fructose (bovine serum albumin), BSA-methylglyoxal, and arginine methylglyoxal models. The pure isolates showed a protective effect in metabolic syndromes as well as promising antioxidant activity. The results showed potent activity of compound 5 in all measured parameters meanwhile, none of the tested compounds showed activity against pancreatic lipase.

Keywords: Chrozophora oblongifolia, antioxidant, pancreatic lipase, metabolic syndromes

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1283 The Effectiveness of Dialectical Behavior Therapy in Developing Emotion Regulation Skill for Adolescent with Intellectual Disability

Authors: Shahnaz Safitri, Rose Mini Agoes Salim, Pratiwi Widyasari

Abstract:

Intellectual disability is characterized by significant limitations in intellectual functioning and adaptive behavior that appears before the age of 18 years old. The prominent impacts of intellectual disability in adolescents are failure to establish interpersonal relationships as socially expected and lower academic achievement. Meanwhile, it is known that emotion regulation skills have a role in supporting the functioning of individual, either by nourishing the development of social skills as well as by facilitating the process of learning and adaptation in school. This study aims to look for the effectiveness of Dialectical Behavior Therapy (DBT) in developing emotion regulation skills for adolescents with intellectual disability. DBT's special consideration toward clients’ social environment and their biological condition is foreseen to be the key for developing emotion regulation capacity for subjects with intellectual disability. Through observations on client's behavior, conducted before and after the completion of DBT intervention program, it was found that there is an improvement in client's knowledge and attitudes related to the mastery of emotion regulation skills. In addition, client's consistency to actually practice emotion regulation techniques over time is largely influenced by the support received from the client's social circles.

Keywords: adolescent, dialectical behavior therapy, emotion regulation, intellectual disability

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1282 A phytochemical and Biological Study of Viscum schemperi Engl. Growing in Saudi Arabia

Authors: Manea A. I. Alqrad, Alaa Sirwi, Sabrin R. M. Ibrahim, Hossam M. Abdallah, Gamal A. Mohamed

Abstract:

Phytochemical study of the methanolic extract of the air dried powdered of the parts of Viscum schemperi Engl. (Family: Viscaceae) using different chromatographic techniques led to the isolation of five compounds: -amyrenone (1), betulinic acid (2), (3β)-olean-12-ene-3,23-diol (3), -oleanolic acid (4), and α-oleanolic acid (5). Their structures were established based on physical, chemical, and spectral data. Anti-inflammatory and anti-apoptotic activities of oleanolic acid in a mouse model of acute hepatorenal damage were assessed. This study showed the efficacy of oleanolic acid to counteract thioacetamide-induced hepatic and kidney injury in mice through the reduction of hepatocyte oxidative damage, suppression of inflammation, and apoptosis. More importantly, oleanolic acid suppressed thioacetamide-induced hepatic and kidney injury by inhibiting NF-κB/TNF-α-mediated inflammation/apoptosis and enhancing SIRT1/Nrf2/Heme-oxygenase signalling pathway. These promising pharmacological activities suggest the potential use of oleanolic acid against hepatorenal damage.

Keywords: oleanolic acid, viscum schimperi, thioacetamide, SIRT1/Nrf2/NF-κB, hepatorenal damage

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1281 The Antibacterial Efficacy of Gold Nanoparticles Derived from Gomphrena celosioides and Prunus amygdalus (Almond) Leaves on Selected Bacterial Pathogens

Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

Abstract:

Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: gold nanoparticles, Gomphrena celesioides, Prunus amygdalus, pathogens

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1280 Combined Optical Coherence Microscopy and Spectrally Resolved Multiphoton Microscopy

Authors: Bjorn-Ole Meyer, Dominik Marti, Peter E. Andersen

Abstract:

A multimodal imaging system, combining spectrally resolved multiphoton microscopy (MPM) and optical coherence microscopy (OCM) is demonstrated. MPM and OCM are commonly integrated into multimodal imaging platforms to combine functional and morphological information. The MPM signals, such as two-photon fluorescence emission (TPFE) and signals created by second harmonic generation (SHG) are biomarkers which exhibit information on functional biological features such as the ratio of pyridine nucleotide (NAD(P)H) and flavin adenine dinucleotide (FAD) in the classification of cancerous tissue. While the spectrally resolved imaging allows for the study of biomarkers, using a spectrometer as a detector limits the imaging speed of the system significantly. To overcome those limitations, an OCM setup was added to the system, which allows for fast acquisition of structural information. Thus, after rapid imaging of larger specimens, navigation within the sample is possible. Subsequently, distinct features can be selected for further investigation using MPM. Additionally, by probing a different contrast, complementary information is obtained, and different biomarkers can be investigated. OCM images of tissue and cell samples are obtained, and distinctive features are evaluated using MPM to illustrate the benefits of the system.

Keywords: optical coherence microscopy, multiphoton microscopy, multimodal imaging, two-photon fluorescence emission

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1279 Biodeterioration and Biodegradation of Historic Parks of UK by Algae

Authors: Syeda Fatima Manzelat

Abstract:

The present study aims to study the groups of algal genera that are responsible for the biodeterioration, biodegradation, and biological pollution of the structures and features of the two historic parks of the UK. Different sites of Campbell Park and Great Linford Manor Park in Milton Keynes are selected to study the morphological, aesthetic, and physical effects of the algal growth. Specimens and swabs were collected mechanically from selected sites. Algal specimens are preserved in Lugol’s solution and labelled with standard information. Photomicrograph analysis of slides using taxonomic keys and visual observation identified algal species that are homogenously and non-homogenously mixed in the aerial, terrestrial, and aquatic habitats. A qualitative study revealed seven classes of Algae. Most of the algal genera isolated have proven records of potential biodegradation, discoloration, and biological pollution. Chlorophyceae was predominantly represented by eleven genera: Chlorella, Chlorococcum Cladophora, Coenochloris Cylindrocapsa, Microspora, Prasiola, Spirogyra, Trentepholia, Ulothrix and Zygnema. Charophyceae is represented by four genera: Cosmarium, Klebsormidium, Mesotaenium, and Mougeotia. Xanthophyceae with two genera, Tribonema and Vaucheria. Bacillariophyceae (Diatoms) are represented by six genera: Acnanthes, Bacillaria, Fragilaria, Gomphonema, Synedra, and Tabellaria, Dinophyceae with a Dinoflagellate. Rhodophyceae included Bangia and Batrachospermum, Cyanophyceae with five genera, Chroococcus, Gloeocapsa, Scytonema, Stigonema and Oscillatoria. The quantitative analysis by statistical method revealed that Chlorophyceae was the predominant class, with eleven genera isolated from different sites of the two parks. Coenochloris of Chlorophyceae was isolated from thirteen sites during the study, followed by Gloeocapsa of Cyanophyceae, which is isolated from 12 sites. These two algae impart varying shades of green colour on the surfaces on which they form biofilms. Prasiola, Vaucheria, and Trentepholia were isolated only from Great Linford Park. Trentepholia imparted a significant orange colour to the walls and trees of the sites. The compounds present in algae that are responsible for discoloration are the green pigment chlorophyll, orange pigment β-carotene, and yellow pigment quinone. Mesotaenium, Dinoflagellate, Gomphonema, Fragilaria, Tabellaria and two unidentified genera were isolated from Campbell Park only. Largest number of algal genera (25) were isolated from the canal of Campbell Park followed by (21) from the canal at Great Linford Manor Park. The Algae were found to grow on surfaces of walls, wooden fencings, metal sculptures, and railings. The Algae are reported to induce surface erosion, natural weathering, and cracking, leading to technical and mechanical instability and extensive damage to building materials. The algal biofilms secrete different organic acids, which are responsible for biosolubilization and biodeterioration of the building materials. The aquatic algal blooms isolated during the study release toxins which are responsible for allergy, skin rashes, vomiting, diarrhea, fever, muscle spasms, and lung and throat infections. The study identifies the places and locations at the historic sites which need to be paid attention. It provides an insight to the conservation strategies to overcome the negative impacts of bio colonization by algae. Prevention measures by different treatments need to be regularly monitored.

Keywords: algae, biodegradation, historic gardens, UK

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1278 Biodeterioration and Biodegradation of Historic Parks of UK by Algae

Authors: Syeda Fatima Manzelat

Abstract:

The present study aims to study the groups of algal genera which are responsible for the biodeterioration, biodegradation and biological pollution of the structures and features of the two historic parks of UK. Different sites of Campbell Park and Great Linford Manor Park in Milton Keynes are selected to study morphological, aesthetic and physical effect of the algal growth. Specimens and swabs were collected mechanically from selected sites. Algal specimens are preserved in Lugol’s solution and labelled with standard information. Photomicrograph analysis of slides using taxonomic keys and visual observation identified algal species that are homogenously and non-homogenously mixed in the aerial, terrestrial and aquatic habitats. Qualitative study revealed seven classes of Algae. Most of the algal genera isolated are with proven records of potential biodegradation, discoloration and biological pollution. Chlorophyceae was predominant represented by eleven genera Chlorella, Chlorococcum Cladophora, Coenochloris Cylindrocapsa. Microspora, Prasiola, Spirogyra, Trentepholia, Ulothrix and Zygnema. Charophyceae is represented by four genera Cosmarium Klebsormidium, Mesotaenium and Mougeotia. Xanthophyceae with two genera Tribonema andVaucheria. Bacillariophyceae (Diatoms) represented by six genera Acnanthes, Bacillaria, Fragilaria, Gomphonema, Synedra and Tabellaria. Dinophyceae with a Dinoflagellate. Rhodophyceae included Bangia and Batrachospermum. Cyanophyceae with five genera, Chroococcus, Gloeocapsa, Scytonema, Stigonema and Oscillatoria. The quantitative analysis by statistical method revealed that Chlorophyceae was the predominant class with eleven genera isolated from different sites of the two parks. Coenochloris of Chlorophyceae was isolated from thirteen sites during the study followed by Gloeocapsa of Cyanophyceae which is isolated from 12 sites. These two algae impart varying shades of green colour on the surfaces on which they form biofilms. Prasiola, Vaucheria and Trentepholia were isolated only from Great Linford Park. Trentepholia imparted a significant orange colour to the walls and trees of the sites. The compounds present in algae that are responsible for discoloration are the green pigment chlorophyll, orange pigment β-carotene and yellow pigment quinone. Mesotaenium, Dinoflagellate, Gomphonema, Fragilaria, Tabellaria and 2 unidentified genera were isolated from Campbell Park only. Largest number of algal genera (25) were isolated from the canal of Campbell Park followed by (21) from the canal at Great Linford Manor Park. The Algae were found to grow on surfaces of walls, wooden fencings, metal sculptures and railing. The Algae are reported to induce surface erosion, natural weathering and cracking leading to technical and mechanical instability and extensive damage to building materials The algal biofilms secrete different organic acids which are responsible for bio solubilization and biodeterioration of the building materials. The aquatic algal blooms isolated during the study release toxins which are responsible for allergy, skin rashes, vomiting, diarrhoea, fever, muscle spasms, lung and throat infections The study identifies the places and locations at the historic sites which need to be paid attention. It provides an insight to the conservation strategies to overcome the negative impacts of bio colonization by algae. Prevention measures by different treatments need to be regularly monitored.

Keywords: algae, biodeterioration, historic gardens, UK

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1277 Development of Electroencephalograph Collection System in Language-Learning Self-Study System That Can Detect Learning State of the Learner

Authors: Katsuyuki Umezawa, Makoto Nakazawa, Manabu Kobayashi, Yutaka Ishii, Michiko Nakano, Shigeichi Hirasawa

Abstract:

This research aims to develop a self-study system equipped with an artificial teacher who gives advice to students by detecting the learners and to evaluate language learning in a unified framework. 'Detecting the learners' means that the system understands the learners' learning conditions, such as each learner’s degree of understanding, the difference in each learner’s thinking process, the degree of concentration or boredom in learning, and problem solving for each learner, which can be interpreted from learning behavior. In this paper, we propose a system to efficiently collect brain waves from learners by focusing on only the brain waves among the biological information for 'detecting the learners'. The conventional Electroencephalograph (EEG) measurement method during learning using a simple EEG has the following disadvantages. (1) The start and end of EEG measurement must be done manually by the experiment participant or staff. (2) Even when the EEG signal is weak, it may not be noticed, and the data may not be obtained. (3) Since the acquired EEG data is stored in each PC, there is a possibility that the time of data acquisition will be different in each PC. This time, we developed a system to collect brain wave data on the server side. This system overcame the above disadvantages.

Keywords: artificial teacher, e-learning, self-study system, simple EEG

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1276 In Silico Design of Organometallic Complexes as Potential Antibacterial Agents

Authors: Sanja O. Podunavac-Kuzmanović, Strahinja Z. Kovačević, Lidija R. Jevrić, Stela Jokić

Abstract:

The complexes of transition metals with various organic ligands have been extensively studied as models of some important pharmaceutical molecules. It was found that biological properties of different substituted organic molecules are improved when they are complexed by different metals. Therefore, it is of great importance for the development of coordination chemistry to explore the assembly of functional organic ligands with metal ion and to investigate the relationship between the structure and property. In the present work, we have bioassayed the antibacterial potency of benzimidazoles and their metal salts (Cu or Zn) against yeast Sarcina lutea. In order to validate our in vitro study, we performed in silico studies using molecular docking software. The investigated compounds and their metal complexes (Cu, Zn) showed good to moderate inhibitory activity against Sarcina lutea. In silico docking studies of the synthesized compounds suggested that complexed benzimidazoles have a greater binding affinity and improved antibacterial activity in comparison with non-complexed ligands. These results are part of the CMST COST Action No. 1105 "Functional metal complexes that bind to biomolecules".

Keywords: organometallic complexes, benzimidazoles, chemometric design, Sarcina lutea

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1275 Effect of Temperatures on Growth and Development Time of Aphis fabae Scopoli (Homoptera: Aphididae): On Bean (Phaseolus vulgaris L.)

Authors: Rochelyn Dona, Serdar Satar

Abstract:

The aim of this study was to evaluate the biological parameters of A. fabae Scopoli (Hemiptera: Aphididae). Developmental, survival, and reproductive data were collected for Aphis fabae reared on detached bean leaves (Phaseolus vulgaris L.) ‘pinto beans’ at five temperature regimes (12, 16, 20, 24, and 28 °C), 65% relative humidity (RH), relative and a photoperiod of 16:8 (LD) h. The developmental times of immature stages ranged from 16, 65 days at 12°C to 5.70 days at 24°C, but a slight increase again at 28°C (6.62 days). At 24°C from this study presented the developmental threshold for A. fabae slightly to 24°C. The average longevity of mature females significantly decreased from 42.32 days at 12°C to 16.12 days at 28°C. The reproduction rate per female was 62.27 at 16°C and 12.72 at 28°C. The mean generation period of the population ranged from 29.24 at 12°C to 11.50 at 28°C. The highest intrinsic rate of increase (rm = 0.41) were recorded at 24°C, the lowest at 12°C (rm = 0.15). It was evident that temperatures over 28°C augmented the development time, accelerated the death ratio of the nymphal stages, Shrunk Adult longevity, and reduced fecundity. The optimal range of temperature for the population growth of A. fabae on the bean was 16°C-24°C, according to this study.

Keywords: developmental time, intrinsic rate, reproduction period, temperature dependence

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1274 The Use of Nano-Crystalline Starch in Probiotic Yogurt and Its Effects on the Physicochemical and Biological Properties

Authors: Ali Seirafi

Abstract:

The purpose of this study was to investigate the effect and application of starch nanocrystals on physicochemical and microbial properties in the industrial production of probiotic yogurt. In this study, probiotic yoghurt was manufactured by industrial method with the optimization and control of the technological factors affecting the probabilistic biomass, using probiotic bacteria Lactobacillus acidophilus and Bifidobacterium bifidum with commonly used yogurt primers. Afterwards, the effects of different levels of fat (1.3%, 2.5 and 4%), as well as the effects of various perbiotic compounds include starch nanocrystals (0.5%, 1 and 1.5%), galactolegalosaccharide (0.5% 1 and 1.5%) and fructooligosaccharide (0.5%, 1 and 1.5%) were evaluated. In addition, the effect of packaging (polyethylene and glass) was studied, while the effect of pH changes and final acidity were studied at each stage. In this research, all experiments were performed in 3 replications and the results were analyzed in a completely randomized design with SAS version 9.1 software. The results of this study showed that the addition of starch nanocrystal compounds as well as the use of glass packaging had the most positive effects on the survival of Lactobacillus acidophilus bacteria and the addition of nano-crystals and the increase in the cooling rate of the product, had the most positive effects on the survival of bacteria Bifidobacterium bifidum.

Keywords: Bifidobacterium bifidum, Lactobacillus acidophilus, prebiotics, probiotic yogurt

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1273 Edward Bond's Questioning of Existence in His Play 'Have I None?'

Authors: Aycan Gurluyer

Abstract:

21st-century theatre has been shaped by lots of world-changing forces devastating human psychology and existence. Accepted as the greatest living English playwright, it is post-war British dramatist Edward Bond who uses a late-21st-century apocalyptic landscape as a weapon to question the human existence in his play 'Have I None?'. In this play, he tries to underline the degenerating and destructive effects of the society and environment on a couple whose lives are changed by an unexpected and annoying stranger. As victim of the society and the cultural corruption, the three vulnerable Bondian characters struggle for their expectations to find a place in this fictional world by sacrificing their own lives. Set in the 2077’s world, the play depicts that rigidly formed rules of the system/authority eliminates the existence of humans. According to Bond, the fascist practices of the governments/systems make people paralyzed in any way, so they choose to deny all realities by becoming biological beings or they gather to commit to suicide as troops. Our main aim is to underscore the questioning of the human existence by drawing the socio-political framework of the era, the capitalist system’s dehumanized individuals and their defence to survive, and what reality is in the 21st century, by focusing on Bond’s hallucinatory and tragic vision of the future in 'Have I None?'.

Keywords: Edward Bond, apocalyptic, existence, Have I None?

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1272 Development and Characterization of Bio-Tribological, Nano- Multilayer Coatings for Medical Tools Application

Authors: L. Major, J. M. Lackner, M. Dyner, B. Major

Abstract:

Development of new generation bio- tribological, multilayer coatings, opens an avenue for fabrication of future high- tech functional surfaces. In the presented work, nano- composite, Cr/CrN+[Cr/ a-C:H implanted by metallic nanocrystals] multilayer coatings have been developed for surface protection of medical tools. Thin films were fabricated by a hybrid Pulsed Laser Deposition technique. Complex microstructure analysis of nano- multilayer coatings, subjected to mechanical and biological tests, were performed by means of transmission electron microscopy (TEM). Microstructure characterization revealed the layered arrangement of Cr23C6 nanoparticles in multilayer structure. Influence of deposition conditions on bio- tribological properties of the coatings were studied. The bio-tests were used as a screening tool for the analyzed nano- multilayer coatings before they could be deposited on medical tools. Bio- medical tests were done using fibroblasts. The mechanical properties of the coatings were investigated by means of a ball-on-disc mechanical test. The microhardness was done using Berkovich indenter. The scratch adhesion test was done using Rockwell indenter. From the bio- tribological point of view, the optimal properties had the C106_1 material.

Keywords: bio- tribological coatings, cell- material interaction, hybrid PLD, tribology

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1271 Discriminant Function Based on Circulating Tumor Cells for Accurate Diagnosis of Metastatic Breast Cancer

Authors: Hatem A. El-Mezayen, Ahmed Abdelmajeed, Fatehya Metwally, Usama Elsaly, Salwa Atef

Abstract:

Tumor metastasis involves the dissemination of malignant cells into the basement membrane and vascular system contributes to the circulating pool of these markers. In this context our aim has been focused on development of a non-invasive. Circulating tumor cells (CTCs) represent a unique liquid biopsy carrying comprehensive biological information of the primary tumor. Herein, we sought to develop a novel score based on the combination of the most significant CTCs biomarkers with and routine laboratory tests for accurate detection of metastatic breast cancer. Methods: Cytokeratin 18 (CK18), Cytokeratin 19 (CK19), and CA15.3 were assayed in metastatic breast cancer (MBC) patients (75), non-MBC patients (50) and healthy control (20). Results: Areas under receiving operating curve (AUCs) were calculated and used for construction on novel score. A novel score named MBC-CTCs = CA15.3 (U/L) × 0.08 + CK 18 % × 2.9 + CK19 × 3.1– 510. That function correctly classified 87% of metastatic breast cancer at cut-off value = 0.55. (i.e great than 0.55 indicates patients with metastatic breast cancer and less than 0.55 indicates patients with non-metastatic breast cancer). Conclusion: MBC-CTCs is a novel, non-invasive and simple can applied to discriminate patients with metastatic breast cancer.

Keywords: metastatic breast cancer, circulating tumor cells, cytokeratin, EpiCam

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1270 Assessment of Relationships between Agro-Morphological Traits and Cold Tolerance in Faba Bean (vicia faba l.) and Wild Relatives

Authors: Nisa Ertoy Inci, Cengiz Toker

Abstract:

Winter or autumn-sown faba bean (Vicia faba L.) is one the most efficient ways to overcome drought since faba bean is usually grown under rainfed where drought and high-temperature stresses are the main growth constraints. The objectives of this study were assessment of (i) relationships between cold tolerance and agro-morphological traits, and (ii) the most suitable agro-morphological trait(s) under cold conditions. Three species of the genus Vicia L. includes 109 genotypes of faba bean (Vicia faba L.), three genotypes of narbon bean (V. narbonensis L.) and two genotypes of V. montbretii Fisch. & C.A. Mey. Davis and Plitmann were sown in autumn at highland of Mediterranean region of Turkey. All relatives of faba bean were more cold-tolerant than the faba bean genotypes. Three faba bean genotypes, ACV-42, ACV-84 and ACV-88, were selected as sources of cold tolerance under field conditions. Path and correlation coefficients and factor and principal component analyses indicated that biological yield should be evaluated in selection for cold tolerance under cold conditions ahead of many agro-morphological traits. The seed weight should be considered for selection in early breeding generations because they had the highest heritability.

Keywords: cold tolerance, faba bean, narbon bean, selection

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1269 Impact of Chimerism on Y-STR DNA Determination: Sex Mismatch Analysis

Authors: Anupuma Raina, Ajay P. Balayan, Prateek Pandya, Pankaj Shrivastava, Uma Kanga, Tulika Seth

Abstract:

DNA fingerprinting analysis aids in personal identification for forensic purposes and has always been a driving motivation for law enforcement agencies in almost all countries since its inception. The introduction of DNA markers (Y-STR) has allowed for greater precision and higher discriminatory power in forensic testing. A criminal/ person committing crime after bone marrow transplantation is a rare situation but not an impossible one. Keeping such a situation in mind, a study was carried out to find out the best biological sample to be used for personal identification, especially in forensic situation. We choose a female patient (recipient) and a male donor. The pre transplant sample (blood) and post transplant samples (blood, buccal swab, hair roots) were collected from the recipient (patient). The same were compared with the blood sample of the donor using DNA FP technique. Post transplant samples were collected at different interval of time (15, 30, 60, and 90 days). The study was carried out using Y-STR kit at 23 loci. The results determined discusses the phenomenon of chimerism and its impact on Y-STR. Hair sample was found the most suitable sample which had no donor DNA profiling up to 90 days.

Keywords: bone marrow transplantation, chimerism, DNA profiling, Y-STR

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1268 Quinazolino-Thiazoles: Fused Pharmacophores as Antimicrobial Agents

Authors: Sanjay Bari, Vinod Ugale, Kamalkishor Patil

Abstract:

Over the past several years the emergence of micro-organisms resistant to nearly all the class of antimicrobial agents has become a serious public health concern. In the present research, we report the synthesis and in-vitro antimicrobial activity of a new series of novel quinazolino-thiadiazoles 3 (a-j). The synthesized compounds were confirmed by melting point, IR, 1H-NMR, 13C NMR and Mass spectroscopy. In general, the results of the in-vitro antibacterial activity are encouraging, as out of 10 compounds tested, Compound 3f and 3i with a 4-chloro phenyl and 4-nitro phenyl at C-2 of thiadiazolyl of quinazolino-thiadiazoles, displayed the excellent antibacterial and antifungal activities against all the tested microorganisms (Bacterial and Fungal strain) with MIC values of 62.5 μg/mL. It is worth to mention that the combination of two biologically active moieties quinazoline and thiadiazole profoundly influences the biological activity. While evaluating the antimicrobial activity, it was observed that compounds having electron withdrawing groups on thiazole has shown profound activity in comparison to compounds having electron releasing groups. As a result of this study, it can be concluded that halogen substituent on thiazole ring increases antimicrobial activity. Possible improvements in the antimicrobial activity can be further achieved by slight modifications in the substituent’s and/or additional structural activity investigations to have good antimicrobial activity.

Keywords: antifungal, antimicrobial, quinazolino-thiazoles, synthesis

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1267 Relevance of Dosing Time for Everolimus Toxicity on Thyroid Gland and Hormones in Mice

Authors: Dilek Ozturk, Narin Ozturk, Zeliha Pala Kara, Engin Kaptan, Serap Sancar Bas, Nurten Ozsoy, Alper Okyar

Abstract:

Most physiological processes oscillate in a rhythmic manner in mammals including metabolism and energy homeostasis, locomotor activity, hormone secretion, immune and endocrine system functions. Endocrine body rhythms are tightly regulated by the circadian timing system. The hypothalamic-pituitary-thyroid (HPT) axis is under circadian control at multiple levels from hypothalamus to thyroid gland. Since circadian timing system controls a variety of biological functions in mammals, circadian rhythms of biological functions may modify the drug tolerability/toxicity depending on the dosing time. Selective mTOR (mammalian target of rapamycin) inhibitor everolimus is an immunosuppressant and anticancer agent that is active against many cancers. It was also found to be active in medullary thyroid cancer. The aim of this study was to investigate the dosing time-dependent toxicity of everolimus on the thyroid gland and hormones in mice. Healthy C57BL/6J mice were synchronized with 12h:12h Light-Dark cycle (LD12:12, with Zeitgeber Time 0 – ZT0 – corresponding to Light onset). Everolimus was administered to male (5 mg/kg/day) and female mice (15 mg/kg/day) orally at ZT1-rest period- and ZT13-activity period- for 4 weeks; body weight loss, clinical signs and possible changes in serum thyroid hormone levels (TSH and free T4) were examined. Histological alterations in the thyroid gland were evaluated according to the following criteria: follicular size, colloid density and viscidity, height of the follicular epithelium and the presence of necrotic cells. The statistical significance between differences was analyzed with ANOVA. Study findings included everolimus-related diarrhea, decreased activity, decreased body weight gains, alterations in serum TSH levels, and histopathological changes in thyroid gland. Decreases in mean body weight gains were more evident in mice treated at ZT1 as compared to ZT13 (p < 0.001, for both sexes). Control tissue sections of thyroid glands exhibited well-organized histoarchitecture when compared to everolimus-treated groups. Everolimus caused histopathological alterations in thyroid glands in male (5 mg/kg, slightly) and female mice (15 mg/kg; p < 0.01 for both ZT as compared to their controls) irrespective of dosing-time. TSH levels were slightly decreased upon everolimus treatment at ZT13 in both males and females. Conversely, increases in TSH levels were observed when everolimus treated at ZT1 in both males (5 mg/kg; p < 0.05) and females (15 mg/kg; slightly). No statistically significant alterations in serum free T4 levels were observed. TSH and free T4 is clinically important thyroid hormones since a number of disease states have been linked to alterations in these hormones. Serum free T4 levels within the normal ranges in the presence of abnormal serum TSH levels in everolimus treated mice may suggest subclinical thyroid disease which may have repercussions on the cardiovascular system, as well as on other organs and systems. Our study has revealed the histological damage on thyroid gland induced by subacute everolimus administration, this effect was irrespective of dosing time. However, based on the body weight changes and clinical signs upon everolimus treatment, tolerability for the drug was best following dosing at ZT13 in both male and females. Yet, effects of everolimus on thyroid functions may deserve further studies regarding their clinical importance and chronotoxicity.

Keywords: circadian rhythm, chronotoxicity, everolimus, thyroid gland, thyroid hormones

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1266 DNpro: A Deep Learning Network Approach to Predicting Protein Stability Changes Induced by Single-Site Mutations

Authors: Xiao Zhou, Jianlin Cheng

Abstract:

A single amino acid mutation can have a significant impact on the stability of protein structure. Thus, the prediction of protein stability change induced by single site mutations is critical and useful for studying protein function and structure. Here, we presented a deep learning network with the dropout technique for predicting protein stability changes upon single amino acid substitution. While using only protein sequence as input, the overall prediction accuracy of the method on a standard benchmark is >85%, which is higher than existing sequence-based methods and is comparable to the methods that use not only protein sequence but also tertiary structure, pH value and temperature. The results demonstrate that deep learning is a promising technique for protein stability prediction. The good performance of this sequence-based method makes it a valuable tool for predicting the impact of mutations on most proteins whose experimental structures are not available. Both the downloadable software package and the user-friendly web server (DNpro) that implement the method for predicting protein stability changes induced by amino acid mutations are freely available for the community to use.

Keywords: bioinformatics, deep learning, protein stability prediction, biological data mining

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1265 Development of Corn (Zea mays L.) Stalk Geotextile Net for Soil Erosion Mitigation

Authors: Cristina S. Decano, Vitaliana U. Malamug, Melissa E. Agulto, Helen F. Gavino

Abstract:

This study aimed to introduce new natural fiber to be used in the production of geotextile net for mitigation of soil erosion. Fiber extraction from the stalks was the main challenge faced during the processing of stalks to ropes. Thus, an investigation on the extraction procedures of corn (Zea mays L.) stalk under biological and chemical retting was undertaken. Results indicated significant differences among percent fiber yield as affected by the retting methods used with values of 15.07%, 12.97%, 11.60%, and 9.01%, for dew, water, chemical (1 day after harvest and15 days after harvest), respectively, with the corresponding average extracting duration of 70, 82, 89, and 94 minutes. Physical characterization of the developed corn stalk geotextile net resulted to average mass per unit area of 806.25 g/m2 and 241% water absorbing capacity. The effect of corn stalk geotextile net in mitigating soil erosion was evaluated in a laboratory experiment for 30o and 60o inclinations with three treatments: bare soil (A1), corn stalk geotextile net (A2) and combined cornstalk geotextile net and vegetation cover (A3). Results revealed that treatment A2 and A3 significantly decreased sediment yield and an increase in terms of soil loss reduction efficiency. The cost of corn stalk geotextile net is Php 62.41 per square meter.

Keywords: corn stalk, natural geotextile, retting, soil erosion

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1264 Studies on Bioaccumulation of 51Cr by Ulva sp. and Ruppia maritima

Authors: Clarissa L. de Araujo, Kátia N. Suzuki, Wilson T. V. Machado, Luis F. Bellido, Alfredo V.B. Bellido

Abstract:

This study aims at contributing to the characterization of the process of biological incorporation of chromium by two benthonic species, the macroalgae Ulva sp. and the aquatic macrophyte Ruppia maritima, to subsidize future activities of monitoring the contamination of aquatic biota. This study is based on laboratory experiments to characterize the incorporation kinetics of the radiotracer 51Cr in two oxidation states (III and VI), under different salinities (7, 15, and 21 ‰). Samples of two benthonic species were collected on the margins of Rodrigo de Freitas Lagoon (Rio de Janeiro, Brazil), acclimated in the laboratory and subsequently subjected to experiments. In tests with 51Cr (III and IV), it was observed that accumulation of the metal in Ulva sp. has inverse relationship with salinity, while for R. maritima, the maximum accumulation occurs in salinity 21‰. In experiments with Cr(III), increases in the uptake of ion by both species were verified. The activity of Cr(III) was up to 19 times greater than the Cr(VI). As regards the potential for accumulation of metals, a better sensitivity of Ulva sp. for any chromium tri or hexavalent forms was verified, while for the Cr(VI) it will require low salinities and longer exposure (>24h). For R. maritima, the results showed the uptake of Cr(VI) increase along with time (>20h), because this species is more resistant for the hexavalent form and useful for any salinity as well.

Keywords: chromium, Cr-51, macroalgae, macrophyte, uptake

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1263 Relevance of Dosing Time for Everolimus Toxicity in Respect to the Circadian P-Glycoprotein Expression in Mdr1a::Luc Mice

Authors: Narin Ozturk, Xiao-Mei Li, Sylvie Giachetti, Francis Levi, Alper Okyar

Abstract:

P-glycoprotein (P-gp, MDR1, ABCB1) is a transmembrane protein acting as an ATP-dependent efflux pump and functions as a biological barrier by extruding drugs and xenobiotics out of cells in healthy tissues especially in intestines, liver and brain as well as in tumor cells. The circadian timing system controls a variety of biological functions in mammals including xenobiotic metabolism and detoxification, proliferation and cell cycle events, and may affect pharmacokinetics, toxicity and efficacy of drugs. Selective mTOR (mammalian target of rapamycin) inhibitor everolimus is an immunosuppressant and anticancer drug that is active against many cancers, and its pharmacokinetics depend on P-gp. The aim of this study was to investigate the dosing time-dependent toxicity of everolimus with respect to the intestinal P-gp expression rhythms in mdr1a::Luc mice using Real Time-Biolumicorder (RT-BIO) System. Mdr1a::Luc male mice were synchronized with 12 h of Light and 12 h of Dark (LD12:12, with Zeitgeber Time 0 – ZT0 – corresponding Light onset). After 1-week baseline recordings, everolimus (5 mg/kg/day x 14 days) was administered orally at ZT1-resting period- and ZT13-activity period- to mdr1a::Luc mice singly housed in an innovative monitoring device, Real Time-Biolumicorder units which let us monitor real-time and long-term gene expression in freely moving mice. D-luciferin (1.5 mg/mL) was dissolved in drinking water. Mouse intestinal mdr1a::Luc oscillation profile reflecting P-gp gene expression and locomotor activity pattern were recorded every minute with the photomultiplier tube and infrared sensor respectively. General behavior and clinical signs were monitored, and body weight was measured every day as an index of toxicity. Drug-induced body weight change was expressed relative to body weight on the initial treatment day. Statistical significance of differences between groups was validated with ANOVA. Circadian rhythms were validated with Cosinor Analysis. Everolimus toxicity changed as a function of drug timing, which was least following dosing at ZT13, near the onset of the activity span in male mice. Mean body weight loss was nearly twice as large in mice treated with 5 mg/kg everolimus at ZT1 as compared to ZT13 (8.9% vs. 5.4%; ANOVA, p < 0.001). Based on the body weight loss and clinical signs upon everolimus treatment, tolerability for the drug was best following dosing at ZT13. Both rest-activity and mdr1a::Luc expression displayed stable 24-h periodic rhythms before everolimus and in both vehicle-treated controls. Real-time bioluminescence pattern of mdr1a revealed a circadian rhythm with a 24-h period with an acrophase at ZT16 (Cosinor, p < 0.001). Mdr1a expression remained rhythmic in everolimus-treated mice, whereas down-regulation was observed in P-gp expression in 2 of 4 mice. The study identified the circadian pattern of intestinal P-gp expression with an unprecedented precision. The circadian timing depending on the P-gp expression rhythms may play a crucial role in the tolerability/toxicity of everolimus. The circadian changes in mdr1a genes deserve further studies regarding their relevance for in vitro and in vivo chronotolerance of mdr1a-transported anticancer drugs. Chronotherapy with P-gp-effluxed anticancer drugs could then be applied according to their rhythmic patterns in host and tumor to jointly maximize treatment efficacy and minimize toxicity.

Keywords: circadian rhythm, chronotoxicity, everolimus, mdr1a::Luc mice, p-glycoprotein

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1262 Production of Insulin Analogue SCI-57 by Transient Expression in Nicotiana benthamiana

Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim

Abstract:

The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.

Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression

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1261 Modeling the Elastic Mean Free Path of Electron Collision with Pyrimidine: The Screen Corrected Additivity Rule Method

Authors: Aouina Nabila Yasmina, Chaoui Zine El Abiddine

Abstract:

This study presents a comprehensive investigation into the elastic mean free path (EMFP) of electrons colliding with pyrimidine, a precursor to the pyrimidine bases in DNA, employing the Screen Corrected Additivity Rule (SCAR) method. The SCAR method is introduced as a novel approach that combines classical and quantum mechanical principles to elucidate the interaction of electrons with pyrimidine. One of the most fundamental properties characterizing the propagation of a particle in the nuclear medium is its mean free path. Knowledge of the elastic mean free path is essential to accurately predict the effects of radiation on biological matter, as it contributes to the distances between collisions. Additionally, the mean free path plays a role in the interpretation of almost all experiments in which an excited electron moves through a solid. Pyrimidine, the precursor of the pyrimidine bases of DNA, has interesting physicochemical properties, which make it an interesting molecule to study from a fundamental point of view. These include a relatively large dipole polarizability and dipole moment and an electronic charge cloud with a significant spatial extension, which justifies its choice in this present study.

Keywords: elastic mean free path, elastic collision, pyrimidine, SCAR

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1260 Controlled Size Synthesis of ZnO and PEG-ZnO NPs and Their Biological Evaluation

Authors: Mahnoor Khan, Bashir Ahmad, Khizar Hayat, Saad Ahmad Khan, Laiba Ahmad, Shumaila Bashir, Abid Ali Khan

Abstract:

The objective of this study was to synthesize the smallest possible size of ZnO NPs using a modified wet chemical synthesis method and to prepare core shell using polyethylene glycol (PEG) as shell material. Advanced and sophisticated techniques were used to confirm the synthesis, size, and shape of these NPs. Rounded, clustered NPs of size 5.343 nm were formed. Both the plain and core shell NPs were tested against MDR bacteria (E. cloacae, E. amnigenus, Shigella, S. odorifacae, Citrobacter, and E. coli). Both of the NPs showed excellent antibacterial properties, whereas E. cloacae showed maximum zone of inhibition of 16 mm, 27 mm, and 32 mm for 500 μg/ml, 1000 μg/ml, and 1500 μg/ml, respectively for plain ZnO NPs and 18 mm, 28 mm and 35 mm for 500 μg/ml, 1000 μg/ml and 1500 μg/ml for core shell NPs. These NPs were also biocompatible on human red blood cells showing little hemolysis of only 4% for 70 μg/ml for plain NPs and 1.5% for 70 μg/ml for core shell NPs. Core shell NPs were highly biocompatible because of the PEG. Their therapeutic effect as photosensitizers in photodynamic therapy (PDT) for cancer treatment was also monitored. The cytotoxicity of ZnO and PEG-ZnO was evaluated using MTT assay. Our results demonstrated that these NPs could generate ROS inside tumor cells after irradiation which in turn initiates an apoptotic pathway leading to cell death hence proving to be an effective candidate for PDT.

Keywords: ZnO, hemolysis, cytotoxiciy assay, photodynamic therapy, antibacterial

Procedia PDF Downloads 116
1259 Electronic Raman Scattering Calibration for Quantitative Surface-Enhanced Raman Spectroscopy and Improved Biostatistical Analysis

Authors: Wonil Nam, Xiang Ren, Inyoung Kim, Masoud Agah, Wei Zhou

Abstract:

Despite its ultrasensitive detection capability, surface-enhanced Raman spectroscopy (SERS) faces challenges as a quantitative biochemical analysis tool due to the significant dependence of local field intensity in hotspots on nanoscale geometric variations of plasmonic nanostructures. Therefore, despite enormous progress in plasmonic nanoengineering of high-performance SERS devices, it is still challenging to quantitatively correlate the measured SERS signals with the actual molecule concentrations at hotspots. A significant effort has been devoted to developing SERS calibration methods by introducing internal standards. It has been achieved by placing Raman tags at plasmonic hotspots. Raman tags undergo similar SERS enhancement at the same hotspots, and ratiometric SERS signals for analytes of interest can be generated with reduced dependence on geometrical variations. However, using Raman tags still faces challenges for real-world applications, including spatial competition between the analyte and tags in hotspots, spectral interference, laser-induced degradation/desorption due to plasmon-enhanced photochemical/photothermal effects. We show that electronic Raman scattering (ERS) signals from metallic nanostructures at hotspots can serve as the internal calibration standard to enable quantitative SERS analysis and improve biostatistical analysis. We perform SERS with Au-SiO₂ multilayered metal-insulator-metal nano laminated plasmonic nanostructures. Since the ERS signal is proportional to the volume density of electron-hole occupation in hotspots, the ERS signals exponentially increase when the wavenumber is approaching the zero value. By a long-pass filter, generally used in backscattered SERS configurations, to chop the ERS background continuum, we can observe an ERS pseudo-peak, IERS. Both ERS and SERS processes experience the |E|⁴ local enhancements during the excitation and inelastic scattering transitions. We calibrated IMRS of 10 μM Rhodamine 6G in solution by IERS. The results show that ERS calibration generates a new analytical value, ISERS/IERS, insensitive to variations from different hotspots and thus can quantitatively reflect the molecular concentration information. Given the calibration capability of ERS signals, we performed label-free SERS analysis of living biological systems using four different breast normal and cancer cell lines cultured on nano-laminated SERS devices. 2D Raman mapping over 100 μm × 100 μm, containing several cells, was conducted. The SERS spectra were subsequently analyzed by multivariate analysis using partial least square discriminant analysis. Remarkably, after ERS calibration, MCF-10A and MCF-7 cells are further separated while the two triple-negative breast cancer cells (MDA-MB-231 and HCC-1806) are more overlapped, in good agreement with the well-known cancer categorization regarding the degree of malignancy. To assess the strength of ERS calibration, we further carried out a drug efficacy study using MDA-MB-231 and different concentrations of anti-cancer drug paclitaxel (PTX). After ERS calibration, we can more clearly segregate the control/low-dosage groups (0 and 1.5 nM), the middle-dosage group (5 nM), and the group treated with half-maximal inhibitory concentration (IC50, 15 nM). Therefore, we envision that ERS calibrated SERS can find crucial opportunities in label-free molecular profiling of complicated biological systems.

Keywords: cancer cell drug efficacy, plasmonics, surface-enhanced Raman spectroscopy (SERS), SERS calibration

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