Search results for: fuel cells

Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

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Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

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Authors: G. Battistoni, F. Collamati, E. De Lucia, R. Faccini, C. Mancini-Terracciano, M. Marafini, I. Mattei, S. Muraro, V. Patera, A. Sarti, A. Sciubba, E. Solfaroli Camillocci, M. Toppi, G. Traini, S. M. Valle, C. Voena

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Accelerated charged particles, mainly protons and carbon ions, are presently used in Particle Therapy (PT) to treat solid tumors. The precision of PT exploiting the charged particle high localized dose deposition in tissues and biological effectiveness in killing cancer cells demands for an online dose monitoring technique, crucial to improve the quality assurance of treatments: possible patient mis-positionings and biological changes with respect to the CT scan could negatively affect the therapy outcome. In PT the beam range confined in the irradiated target can be monitored thanks to the secondary radiation produced by the interaction of the projectiles with the patient tissue. The Dose Profiler (DP) is a novel device designed to track charged secondary particles and reconstruct their longitudinal emission distribution, correlated to the Bragg peak position. The feasibility of this approach has been demonstrated by dedicated experimental measurements. The DP has been developed in the framework of the INSIDE project, MIUR, INFN and Centro Fermi, Museo Storico della Fisica e Centro Studi e Ricerche 'E. Fermi', Roma, Italy and will be tested at the Proton Therapy center of Trento (Italy) within the end of 2017. The DP combines a tracker, made of six layers of two-view scintillating fibers with square cross section (0.5 x 0.5 mm2) with two layers of two-view scintillating bars (section 12.0 x 0.6 mm2). The electronic readout is performed by silicon photomultipliers. The sensitive area of the tracking planes is 20 x 20 cm2. To optimize the detector layout, a Monte Carlo (MC) simulation based on the FLUKA code has been developed. The complete DP geometry and the track reconstruction code have been fully implemented in the MC. In this contribution, the DP hardware will be described. The expected detector performance computed using a dedicated simulation of a 220 MeV/u carbon ion beam impinging on a PMMA target will be presented, and the result will be discussed in the standard clinical application framework. A possible procedure for real-time beam range monitoring is proposed, following the expectations in actual clinical operation.

Keywords: online range monitoring, particle therapy, quality assurance, tracking detector

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Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

Procedia PDF Downloads 141

Authors: Santi Maneewatchararangsri, Wanpen Chaicumpa, Patcharin Saengjaruk, Urai Chaisri

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Leptospirosis is a globally neglected disease that continues to be a significant public health and veterinary burden, with millions of cases reported each year. Early and accurate differential diagnosis of leptospirosis from other febrile illnesses and the development of a broad spectrum of leptospirosis vaccines are needed. The LipL32 outer membrane lipoprotein is a member of Leptospira adhesive matrices and has been found to exert hemolytic activity to erythrocytes in vitro. Therefore, LipL32 is regarded as a potential target for diagnosis, broad-spectrum leptospirosis vaccines, and for passive immunotherapy. In this study, we established LipL32-specific mouse monoclonal antibodies, mAbLPF1 and mAbLPF2, and their respective mouse- and humanized-engineered single chain variable fragment (ScFv). Their antibodies’ neutralizing activities against Leptospira-mediated hemolysis in vitro, and the therapeutic efficacy of mAbs against heterologous Leptospira infected hamsters were demonstrated. The epitope peptide of mAb LPF1 was mapped to a non-contiguous carboxy-terminal β-turn and amphipathic α-helix of LipL32 structure contributing to phospholipid/host cell adhesion and membrane insertion. We found that the mAbLPF2 epitope was located on the interacting loop of peptide binding groove of the LipL32 molecule responsible for interactions with host constituents. Epitope sequences are highly conserved among Leptospira spp. and are absent from the LipL32 superfamily of other microorganisms. Both epitopes are surface-exposed, readily accessible by mAbs, and immunogenic. However, they are less dominant when revealed by LipL32-specific immunoglobulins from leptospirosis-patient sera and rabbit hyperimmune serum raised by whole Leptospira. Our study also demonstrated an adhesion inhibitory activity of LipL32 protein to host membrane components and cells mediated by mAbs as well as an anti-hemolytic activity of the respective antibodies. The therapeutic antibodies, particularly the humanized-ScFv, have a potential for further development as non-drug therapeutic agent for human leptospirosis, especially in subjects allergic to antibiotics. The epitope peptides recognized by two therapeutic mAbs have potential use as tools for structure-function studies. Finally, protective peptides may be used as a target for epitope-based vaccines for control of leptospirosis.

Keywords: leptospira lipl32-specific antibodies, therapeutic epitopes, epitopes characterization, immunotherapy

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Authors: Bhupendra Kumar Poudel, Sadhana Amatya, Tirtha Maiya Shrestha, Bharatmani Pokhrel, Mohan Prasad Amatya

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Background: H. salicifolia is a dense, branched, multipurpose, deciduous, nitrogen fixing, thorny willow-like small to moderate tree, restricted to the Himalaya. Among the two species of Nepal (Hippophae salicifolia and H. tibetana), it has been traditionally used as food additive, anticancer (bark), and treating toothache, tooth inflammation (anti-inflammatory) and radiation injury; while people of Western Nepal have largely undermined its veiled treasure by using it for fuel, wood and soil stabilization only. Therefore, the main objective of this study was to explore biological properties (analgesic, antidiabetic, cytotoxic and anti-inflammatory properties of this plant. Methodology: The transverse section of leaves and stems were viewed under microscope. Extracts obtained from soxhlation subjected to tests for phytochemical and biological studies. Rats (used to study antidiabetic and anti-inflammatory properties) and mice (used to study analgesic, CNS depressant, muscle relaxant and locomotor properties) were assumed to be normally distributed; then ANOVA and post hoc tukey test was used to find significance. The data obtained were analyzed by SPSS 17 and Excel 2007. Results and Conclusion: Pharmacognostical analysis revealed the presence of long stellate trichomes, double layered vascular bundle 5-6 in number and double layered compact sclerenchyma. The preliminary phytochemical screening of the extracts was found to exhibit the positive reaction tests for glycoside, steroid, tannin, flavonoid, saponin, coumarin and reducing sugar. The brine shrimp lethality bioassay tested in 1000, 100 and 10 ppm revealed cytotoxic activity inherent in methanol, water, chloroform and ethyl acetate extracts with LC50 (μg/ml) values of 61.42, 99.77, 292.72 and 277.84 respectively. The cytotoxic activity may be due to presence of tannins in the constituents. Antimicrobial screening of the extracts by cup diffusion method using Staphylococcus aereus, Escherichia coli and Pseudomonas aeruginosa against standard antibiotics (oxacillin, gentamycin and amikacin respectively) portrayed no activity against the microorganisms tested. The methanol extract of the stems and leaves showed various pharmacological properties: and antidiabetic, anti-inflammatory, analgesic [chemical writhing method], CNS depressant, muscle relaxant and locomotor activities in a dose-dependent fashion, indicating the possibility of the presence of different constituents in the stems and leaves responsible for these biological activities. All the effects when analyzed by post hoc tukey test were found to be significant at 95% confidence level. The antidiabetic activity was presumed to be due to flavonoids present in extract. Therefore, it can be concluded that this plant’s secondary metabolites possessed strong antidiabetic, anti-inflammatory and cytotoxic activity which could be isolated for further investigation.

Keywords: Hippophae salicifolia, constituents, antidiabetic, inflammatory, brine shrimp

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Authors: Jos Olijve

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Introduction and Purpose: Endotoxins are found in the outer membrane of gram-negative bacteria and have profound in vitro and in vivo responses. They can trigger strong immune responses and negatively affect various cellar activities particular cells expressing toll-like receptors. They are therefore unwanted contaminants of biomaterials sourced from natural raw materials, and their activity must be as low as possible. Collagen and gelatin are natural extracellular matrix components and have, due to their low allergenic potential, suitable biological properties, and tunable physical characteristics, high potential in biomedical applications. The purpose of this study was to determine the influence of autoclave sterilization of gelatin on physical properties and endotoxin level compared to surfactant purified gelatin. Methods: Type A gelatin from Sigma-Aldrich (G1890) with endotoxin level of 35000 endotoxin units (EU) per gram gelatin and type A gelatins from Rousselot Gent with endotoxin activity of 30000 EU per gram were used. A 10 w/w% G1890 gelatin solution was autoclave sterilized during 30 minutes at 121°C and 1 bar over pressure. The physical properties and the endotoxin level of the sterilized G1890 gelatin were compared to a type A gelatin from Rousselot purified with Triton X100 surfactant. The Triton X100 was added to a concentration of 0.5 w/w% which is above the critical micellar concentration. The gelatin surfactant mixtures were kept for 30-45 minutes under constant stirring at 55-60°C. The Triton X100 was removed by active carbon filtration. The endotoxin levels of the gelatins were measured using the Endozyme recombinant factor C method from Hyglos GmbH (Germany). Results and Discussion: Autoclave sterilization significantly affect the physical properties of gelatin. Molecular weight of G1890 decreased from 140 to 50kDa, and gel strength decreased from 300 to 40g. The endotoxin level of the gelatin reduced after sterilization from 35000 EU/g to levels of 400-500 EU/g. These endotoxin levels are however still far above the upper endotoxin level of 0.05 EU/ml, which resembles 5 EU/g gelatin based on a 1% gelatin solution, to avoid cell proliferation alteration. Molecular weight and gel strength of Rousselot gelatin was not altered after Triton X100 purification and remained 150kDa and 300g respectively. The endotoxin levels of Triton X100 purified Rousselot gelatin was < 5EU/g gelatin. Conclusion: Autoclave sterilization of gelatin is, in comparison to Triton X100 purification, not efficient to inactivate endotoxin levels in gelatin to levels below the upper limit to avoid cell proliferation alteration. Autoclave sterilization gave a significant decrease in molecular weight and gel strength which makes autoclave sterilized gelatin, in comparison to Triton X100 purified gelatin, not suitable for 3D printing.

Keywords: endotoxin, gelatin, molecular weight, sterilization, Triton X100

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Authors: Olga Kokoulina

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Today, algorithms are assuming a leading role in various areas of decision-making. Prompted by a promise to provide increased economic efficiency and fuel solutions for pressing societal challenges, algorithmic decision-making is often celebrated as an impartial and constructive substitute for human adjudication. But in the face of this implied objectivity and efficiency, the application of algorithms is also marred with mounting concerns about embedded biases, discrimination, and exclusion. In Europe, vigorous debates on risks and adverse implications of algorithmic decision-making largely revolve around the potential of data protection laws to tackle some of the related issues. For example, one of the often-cited venues to mitigate the impact of potentially unfair decision-making practice is a so-called 'right to explanation'. In essence, the overall right is derived from the provisions of the General Data Protection Regulation (‘GDPR’) ensuring the right of data subjects to access and mandating the obligation of data controllers to provide the relevant information about the existence of automated decision-making and meaningful information about the logic involved. Taking corresponding rights and obligations in the context of the specific provision on automated decision-making in the GDPR, the debates mainly focus on efficacy and the exact scope of the 'right to explanation'. In essence, the underlying logic of the argued remedy lies in a transparency imperative. Allowing data subjects to acquire as much knowledge as possible about the decision-making process means empowering individuals to take control of their data and take action. In other words, forewarned is forearmed. The related discussions and debates are ongoing, comprehensive, and, often, heated. However, they are also frequently misguided and isolated: embracing the data protection law as ultimate and sole lenses are often not sufficient. Mandating the disclosure of technical specifications of employed algorithms in the name of transparency for and empowerment of data subjects potentially encroach on the interests and rights of IPR holders, i.e., business entities behind the algorithms. The study aims at pushing the boundaries of the transparency debate beyond the data protection regime. By systematically analysing legal requirements and current judicial practice, it assesses the limits of the transparency requirement and right to access posed by intellectual property law, namely by copyrights and trade secrets. It is asserted that trade secrets, in particular, present an often-insurmountable obstacle for realising the potential of the transparency requirement. In reaching that conclusion, the study explores the limits of protection afforded by the European Trade Secrets Directive and contrasts them with the scope of respective rights and obligations related to data access and portability enshrined in the GDPR. As shown, the far-reaching scope of the protection under trade secrecy is evidenced both through the assessment of its subject matter as well as through the exceptions from such protection. As a way forward, the study scrutinises several possible legislative solutions, such as flexible interpretation of the public interest exception in trade secrets as well as the introduction of the strict liability regime in case of non-transparent decision-making.

Keywords: algorithms, public interest, trade secrets, transparency

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Authors: Ayman Hijazi, Witold Kwapinski, J. J. Leahy

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Finding a sustainable alternative energy to fossil fuel is an urgent need as various environmental challenges in the world arise. Therefore, formic acid (FA) decomposition has been an attractive field that lies at the center of the biomass platform, comprising a potential pool of hydrogen energy that stands as a distinct energy vector. Liquid FA features considerable volumetric energy density of 6.4 MJ/L and a specific energy density of 5.3 MJ/Kg that qualifies it in the prime seat as an energy source for transportation infrastructure. Additionally, the increasing research interest in FA decomposition is driven by the need for in-situ H₂ production, which plays a key role in the hydrogenation reactions of biomass into higher-value components. It is reported elsewhere in the literature that catalytic decomposition of FA is usually performed in poorly designed setups using simple glassware under magnetic stirring, thus demanding further energy investment to retain the used catalyst. Our work suggests an approach that integrates designing a distinct catalyst featuring magnetic properties with a robust setup that minimizes experimental & measurement discrepancies. One of the most prominent active species for the dehydrogenation/hydrogenation of biomass compounds is palladium. Accordingly, we investigate the potential of engrafting palladium metal onto functionalized magnetic nanoparticles as a heterogeneous catalyst to favor the production of CO-free H₂ gas from FA. Using an ordinary magnet to collect the spent catalyst renders core-shell magnetic nanoparticles as the backbone of the process. Catalytic experiments were performed in a jacketed batch reactor equipped with an overhead stirrer under an inert medium. Through a distinct approach, FA is charged into the reactor via a high-pressure positive displacement pump at steady-state conditions. The produced gas (H₂+CO₂) was measured by connecting the gas outlet to a measuring system based on the amount of the displaced water. The uniqueness of this work lies in designing a very responsive catalyst, pumping a consistent amount of FA into a sealed reactor running at steady-state mild temperatures, and continuous gas measurement, along with collecting the used catalyst without the need for centrifugation. Catalyst characterization using TEM, XRD, SEM, and CHN elemental analyzer provided us with details of catalyst preparation and facilitated new venues to alter the nanostructure of the catalyst framework. Consequently, the introduction of amine groups has led to appreciable improvements in terms of dispersion of the doped metals and eventually attaining nearly complete conversion (100%) of FA after 7 hours. The relative importance of the process parameters such as temperature (35-85°C), stirring speed (150-450rpm), catalyst loading (50-200mgr.), and Pd doping ratio (0.75-1.80wt.%) on gas yield was assessed by a Taguchi design-of-experiment based model. Experimental results showed that operating at a lower temperature range (35-50°C) yielded more gas, while the catalyst loading and Pd doping wt.% were found to be the most significant factors with P-values 0.026 & 0.031, respectively.

Keywords: formic acid decomposition, green catalysis, hydrogen, mesoporous silica, process optimization, nanoparticles

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Authors: Salma A. Mahmoud

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Epigenetic, or alteration in gene expression influenced by extragenetic factors, has emerged as one of the most promising areas that will address some of the gaps in our current knowledge in understanding patterns of human variation. In the last decade, the research investigating epigenetic mechanisms in many fields has flourished and witnessed significant progress. It paved the way for a new era of integrated research especially between anthropology/archeology and life sciences. Skeletal remains are considered the most significant source of information for studying human variations across history, and by utilizing these valuable remains, we can interpret the past events, cultures and populations. In addition to archeological, historical and anthropological importance, studying bones has great implications in other fields such as medicine and science. Bones also can hold within them the secrets of the future as they can act as predictive tools for health, society characteristics and dietary requirements. Bones in their basic forms are composed of cells (osteocytes) that are affected by both genetic and environmental factors, which can only explain a small part of their variability. The primary objective of this project is to examine the epigenetic landscape/signature within bones of archeological remains as a novel marker that could reveal new ways to conceptualize chronological events, gender differences, social status and ecological variations. We attempted here to address discrepancies in common variants such as methylome as well as novel epigenetic regulators such as chromatin remodelers, which to our best knowledge have not yet been investigated by anthropologists/ paleoepigenetists using plethora of techniques (biological, computational, and statistical). Moreover, extracting epigenetic information from bones will highlight the importance of osseous material as a vector to study human beings in several contexts (social, cultural and environmental), and strengthen their essential role as model systems that can be used to investigate and construct various cultural, political and economic events. We also address all steps required to plan and conduct an epigenetic analysis from bone materials (modern and ancient) as well as discussing the key challenges facing researchers aiming to investigate this field. In conclusion, this project will serve as a primer for bioarcheologists/anthropologists and human biologists interested in incorporating epigenetic data into their research programs. Understanding the roles of epigenetic mechanisms in bone structure and function will be very helpful for a better comprehension of their biology and highlighting their essentiality as interdisciplinary vectors and a key material in archeological research.

Keywords: epigenetics, archeology, bones, chromatin, methylome

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Authors: A. Mulry, C. Kealey, D. B. Brady

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Planktonic bacteria can form biofilms which are microbial aggregates embedded within a matrix of extracellular polymeric substances (EPS). They can be found attached to abiotic or biotic surfaces. Biofilms are responsible for oral diseases such as dental caries, gingivitis and the progression of periodontal disease. Biofilms can resist 500 to 1000 times the concentration of biocides and antibiotics used to kill planktonic bacteria. Biofilm development on oral surfaces involves four stages, initial attachment, early development, maturation and dispersal of planktonic cells. The Minimum Inhibitory Concentration (MIC) was determined using a range of saturated and unsaturated fatty acids using the resazurin assay, followed by serial dilution and spot plating on BHI agar plates to establish the Minimum Bactericidal Concentration (MBC). Log reduction of bacteria was also evaluated for each fatty acid. The Minimum Biofilm Inhibition Concentration (MBIC) was determined using crystal violet assay in 96 well plates on forming and pre-formed S. mutans biofilms using BHI supplemented with 1% sucrose. Saturated medium-chain fatty acids Octanoic (C8.0), Decanoic (C10.0) and Undecanoic acid (C11.0) do not display strong antibiofilm properties; however, Lauric (C12.0) and Myristic (C14.0) display moderate antibiofilm properties with 97.83% and 97.5% biofilm inhibition with 1000 µM respectively. Monounsaturated, Oleic acid (C18.1) and polyunsaturated large chain fatty acids, Linoleic acid (C18.2) display potent antibiofilm properties with biofilm inhibition of 99.73% at 125 µM and 100% at 65.5 µM, respectively. Long-chain polyunsaturated Omega-3 fatty acids α-Linoleic (C18.3), Eicosapentaenoic Acid (EPA) (C20.5), Docosahexaenoic Acid (DHA) (C22.6) have displayed strong antibiofilm efficacy from concentrations ranging from 31.25-250µg/ml. DHA is the most promising antibiofilm agent with an MBIC of 99.73% with 15.625µg/ml. This may be due to the presence of six double bonds and the structural orientation of the fatty acid. To conclude, fatty acids displaying the most antimicrobial activity appear to be medium or long-chain unsaturated fatty acids containing one or more double bonds. Most promising agents include Omega-3-fatty acids Linoleic, α-Linoleic, EPA and DHA, as well as Omega-9 fatty acid Oleic acid. These results indicate that fatty acids have the potential to be used as antimicrobials and antibiofilm agents against S. mutans. Future work involves further screening of the most potent fatty acids against a range of bacteria, including Gram-positive and Gram-negative oral pathogens. Future work will involve incorporating the most effective fatty acids onto dental implant devices to prevent biofilm formation.

Keywords: antibiofilm, biofilm, fatty acids, S. mutans

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Authors: Srinivas Bathini, Duraichelvan Raju, Simona Badilescu, Muthukumaran Packirisamy

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A bio-sensing method, based on the plasmonic property of gold nano-islands, has been developed for detection of exosomes in a clinical setting. The position of the gold plasmon band in the UV-Visible spectrum depends on the size and shape of gold nanoparticles as well as on the surrounding environment. By adsorbing various chemical entities, or binding them, the gold plasmon band will shift toward longer wavelengths and the shift is proportional to the concentration. Exosomes transport cargoes of molecules and genetic materials to proximal and distal cells. Presently, the standard method for their isolation and quantification from body fluids is by ultracentrifugation, not a practical method to be implemented in a clinical setting. Thus, a versatile and cutting-edge platform is required to selectively detect and isolate exosomes for further analysis at clinical level. The new sensing protocol, instead of antibodies, makes use of a specially synthesized polypeptide (Vn96), to capture and quantify the exosomes from different media, by binding the heat shock proteins from exosomes. The protocol has been established and optimized by using a glass substrate, in order to facilitate the next stage, namely the transfer of the protocol to a microfluidic environment. After each step of the protocol, the UV-Vis spectrum was recorded and the position of gold Localized Surface Plasmon Resonance (LSPR) band was measured. The sensing process was modelled, taking into account the characteristics of the nano-island structure, prepared by thermal convection and annealing. The optimal molar ratios of the most important chemical entities, involved in the detection of exosomes were calculated as well. Indeed, it was found that the results of the sensing process depend on the two major steps: the molar ratios of streptavidin to biotin-PEG-Vn96 and, the final step, the capture of exosomes by the biotin-PEG-Vn96 complex. The microfluidic device designed for sensing of exosomes consists of a glass substrate, sealed by a PDMS layer that contains the channel and a collecting chamber. In the device, the solutions of linker, cross-linker, etc., are pumped over the gold nano-islands and an Ocean Optics spectrometer is used to measure the position of the Au plasmon band at each step of the sensing. The experiments have shown that the shift of the Au LSPR band is proportional to the concentration of exosomes and, thereby, exosomes can be accurately quantified. An important advantage of the method is the ability to discriminate between exosomes having different origins.

Keywords: exosomes, gold nano-islands, microfluidics, plasmonic biosensing

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Authors: Jara Kaye S. Villanueva, M. Rosario Concepcion O. Ang

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For the past few years, Philippines has depended most of its energy source on oil, coal, and fossil fuel. According to the Department of Energy (DOE), the dominance of coal in the energy mix will continue until the year 2020. The expanding energy needs in the country have led to increasing efforts to promote and develop renewable energy. This research is a part of the government initiative in preparation for renewable energy development and expansion in the country. The Philippine Renewable Energy Resource Mapping from Light Detection and Ranging (LiDAR) Surveys is a three-year government project which aims to assess and quantify the renewable energy potential of the country and to put them into usable maps. This study focuses on the site suitability analysis of the four renewable energy sources – biomass (coconut, corn, rice, and sugarcane), hydro, solar, and wind energy. The site assessment is a key component in determining and assessing the most suitable locations for the construction of renewable energy power plants. This method maximizes the use of both the technical methods in resource assessment, as well as taking into account the environmental, social, and accessibility aspect in identifying potential sites by utilizing and integrating two different methods: the Multi-Criteria Decision Analysis (MCDA) method and Geographic Information System (GIS) tools. For the MCDA, Analytical Hierarchy Processing (AHP) is employed to determine the parameters needed for the suitability analysis. To structure these site suitability parameters, various experts from different fields were consulted – scientists, policy makers, environmentalists, and industrialists. The need to have a well-represented group of people to consult with is relevant to avoid bias in the output parameter of hierarchy levels and weight matrices. AHP pairwise matrix computation is utilized to derive weights per level out of the expert’s gathered feedback. Whereas from the threshold values derived from related literature, international studies, and government laws, the output values were then consulted with energy specialists from the DOE. Geospatial analysis using GIS tools translate this decision support outputs into visual maps. Particularly, this study uses Euclidean distance to compute for the distance values of each parameter, Fuzzy Membership algorithm which normalizes the output from the Euclidean Distance, and the Weighted Overlay tool for the aggregation of the layers. Using the Natural Breaks algorithm, the suitability ratings of each of the map are classified into 5 discrete categories of suitability index: (1) not suitable (2) least suitable, (3) suitable, (4) moderately suitable, and (5) highly suitable. In this method, the classes are grouped based on the best groups similar values wherein each subdivision are set from the rest based on the big difference in boundary values. Results show that in the entire Philippine area of responsibility, biomass has the highest suitability rating with rice as the most suitable at 75.76% suitability percentage, whereas wind has the least suitability percentage with score 10.28%. Solar and Hydro fall in the middle of the two, with suitability values 28.77% and 21.27%.

Keywords: site suitability, biomass energy, hydro energy, solar energy, wind energy, GIS

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Authors: Hamid Abdulroof Saleh

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Background: Dioxins are one of the most widely distributed environmental pollutants. Dioxins consist of feedstock during the preparation of some industries, such as the paper industry as they can be produced in the atmosphere during the process of burning garbage and waste, especially medical waste. Dioxins can be found in the adipose tissues of animals in the food chain as well as in human breast milk. 2,3,7,8-Tetrachlorodibenzo-pdioxin (TCDD) is the most toxic component of a large group of dioxins. Humans are exposed to TCDD through contaminated food items like meat, fish, milk products, eggs etc. Recently, natural formulations relating to reducing or eliminating TCDD toxicity have been in focus. Ginger rhizome (Zingiber officinale R., family: Zingiberaceae), is used worldwide as a spice. Both antioxidative and androgenic activity of Z. officinale was reported in animal models. Researchers showed that ginger oil has dominative protective effect on DNA damage and might act as a scavenger of oxygen radical and might be used as an antioxidant. Aim of the work: The present study was undertaken to evaluate the toxic effect of TCDD on the structure and histoarchitecture of the testis and the protective role of co-administration of ginger root extract to prevent this toxicity. Materials & Methods: Male adult rats of Sprague-Dawley strain were assigned to four groups, eight rats in each; control group, dioxin treated group (given TCDD at the dose of 100 ng/kg Bwt/day by gavage), ginger treated group (given 50 mg/kg Bwt/day of ginger root extract by gavage), dioxin and ginger treated group (given TCDD at the dose of 100 ng/kg Bwt/day and 50 mg/kg Bwt/day of ginger root extract by gavages). After three weeks, rats were weighed and sacrificed where testis were removed and weighted. The testes were processed for routine paraffin embedding and staining. Tissue sections were examined for different morphometric and histopathological changes. Results: Dioxin administration showed a harmful effects in the body, testis weight and other morphometric parameters of the testis. In addition, it produced varying degrees of damage to the seminiferous tubules, which were shrunken and devoid of mature spermatids. The basement membrane was disorganized with vacuolization and loss of germinal cells. The co-administration of ginger root extract showed obvious improvement in the above changes and showed reversible morphometric and histopathological changes of the seminiferous tubules. Conclusion: Ginger root extract treatment in this study was successful in reversing all morphometric and histological changes of dioxin testicular damage. Therefore, it showed a protective effect on testis against dioxin toxicity.

Keywords: dioxin, ginger, rat, testis

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Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

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Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls, and the parasite has developed resistance to nearly all control chemicals used in the past. It is, therefore, critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi, and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: lucilia cuprina, primary cell line establishment, RNA interference, insect cell transfection

Procedia PDF Downloads 53

Authors: Antoine Dumont, Weiji Hong, Zheng-Hong Lu

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Light emitting diodes (LEDs) are steadily becoming the new standard for luminescent display devices because of their energy efficiency and relatively low cost, and the purity of the light they emit. Our research focuses on the optical properties of the lead halide perovskite CsPbBr₃ and its family that is showing steadily improving performances in LEDs and solar cells. The objective of this work is to investigate CsPbBr₃ as an emitting layer made by physical vapor deposition instead of the usual solution-processed perovskites, for use in LEDs. The deposition in vacuum eliminates any risk of contaminants as well as the necessity for the use of chemical ligands in the synthesis of quantum dots. Initial results show the versatility of the dual-source evaporation method, which allowed us to create different phases in bulk form by altering the mole ratio or deposition rate of CsBr and PbBr₂. The distinct phases Cs₄PbBr₆, CsPbBr₃ and CsPb₂Br₅ – confirmed through XPS (x-ray photoelectron spectroscopy) and X-ray diffraction analysis – have different optical properties and morphologies that can be used for specific applications in optoelectronics. We are particularly focused on the blue shift expected from quantum dots (QDs) and the stability of the perovskite in this form. We already obtained proof of the formation of QDs through our dual source evaporation method with electron microscope imaging and photoluminescence testing, which we understand is a first in the community. We also incorporated the QDs in an LED structure to test the electroluminescence and the effect on performance and have already observed a significant wavelength shift. The goal is to reach 480nm after shifting from the original 528nm bulk emission. The hole transport layer (HTL) material onto which the CsPbBr₃ is evaporated is a critical part of this study as the surface energy interaction dictates the behaviour of the QD growth. A thorough study to determine the optimal HTL is in progress. A strong blue shift for a typically green emitting material like CsPbBr₃ would eliminate the necessity of using blue emitting Cl-based perovskite compounds and could prove to be more stable in a QD structure. The final aim is to make a perovskite QD LED with strong blue luminescence, fabricated through a dual-source evaporation technique that could be scalable to industry level, making this device a viable and cost-effective alternative to current commercial LEDs.

Keywords: material physics, perovskite, light emitting diode, quantum dots, high vacuum deposition, thin film processing

Procedia PDF Downloads 144

Authors: Sonia Tamanna, Akramul Hassan, Mohammad Shakil Mahmood, Farzana Ansari, Gowhar Rashid, Mir Fahim Faisal, M. Zakir Hossain Howlader

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Background: Preeclampsia (PE), a pregnancy-specific hypertensive disorder, significantly impacts maternal and fetal health. It is particularly prevalent in underdeveloped countries and is linked to preterm delivery and fetal growth. The renin-angiotensin system (RAS) plays a crucial role in ensuring a successful pregnancy outcome, with Angiotensin-Converting Enzyme 2 (ACE2) being a key component. ACE2 converts ANG II to Ang-(1-7), offering protection against ANG II-induced stress and inflammation while regulating blood pressure and osmotic balance during pregnancy. The reduced maternal plasma angiotensin-converting enzyme 2 (ACE2) seen in preeclampsia might contribute to its pathogenesis. However, there has been a dearth of comprehensive research into the association between ACE2 gene polymorphism and preeclampsia. In the South Asian population, hypertension is strongly linked to two SNPs: rs2285666 and rs879922. This genotype was therefore considered, and the possible association of maternal and fetal ACE2 gene polymorphism with preeclampsia within the Bangladeshi population was evaluated. Method: DNA was extracted from peripheral white blood cells (WBCs) using the organic method, and SNP genotyping was done via PCR-RFLP. Odds ratios (OR) with 95% confidence intervals (95% CI) were calculated using logistic regression to determine relative risk. Result: A comprehensive case-control study was conducted on 51 PE patients and their infants, along with 56 control subjects and their infants. Maternal single nuvleotide polymorphisms (SNP) (rs2285666) analysis revealed a strong association between the TT genotype and preeclampsia, with a four-fold increased risk in mothers (P=0.024, OR=4.00, 95% CI=1.36-11.37) compared to their ancestral genotype CC. However, the CT genotype (rs2285666) showed no significant difference (P=0.46, OR=1.54, 95% CI=0.57-4.14). Notably, no significant correlation was found in infants, regardless of their gender. For rs879922, no significant association was observed in both mothers and infants. This pioneering study suggests that mothers carrying the ACE2 gene variant rs2285666 (TT allele) may be at higher risk for preeclampsia, potentially influencing hypertension characteristics, whereas rs879922 does not appear to be associated with developing preeclampsia. Conclusion: This study sheds light on the role of ACE2 gene polymorphism, particularly the rs2285666 TT allele, in maternal susceptibility to preeclampsia. However, rs879922 does not appear to be linked to the risk of PE. This research contributes to our understanding of the genetic underpinnings of preeclampsia, offering insights into potential avenues for prevention and management.

Keywords: ACE2, PCR-RFLP, preeclampsia, single nuvleotide polymorphisms (SNPs)

Procedia PDF Downloads 43

Authors: Kerekoppa Puttaiah Bhatta Ramesha, Uday Kannegundla, Praseeda Mol, Lathika Gopalakrishnan, Jagish Kour Reen, Gourav Dey, Manish Kumar, Sakthivel Jeyakumar, Arumugam Kumaresan, Kiran Kumar M., Thottethodi Subrahmanya Keshava Prasad

Abstract:

Spermatozoa are the highly specialized transcriptionally and translationally inactive haploid male gamete. The understanding of proteome of sperm is indispensable to explore the mechanism of sperm motility and fertility. Though there is a large number of human sperm proteomic studies, in-depth proteomic information on Bos indicus spermatozoa is not well established yet. Therefore, we illustrated the profile of sperm proteome in indigenous cattle, Malnad gidda (Bos Indicus), using high-resolution mass spectrometry. In the current study, two semen ejaculates from 3 breeding bulls were collected employing the artificial vaginal method. Using 45% percoll purification, spermatozoa cells were isolated. Protein was extracted using lysis buffer containing 2% Sodium Dodecyl Sulphate (SDS) and protein concentration was estimated. Fifty micrograms of protein from each individual were pooled for further downstream processing. Pooled sample was fractionated using SDS-Poly Acrylamide Gel Electrophoresis, which is followed by in-gel digestion. The peptides were subjected to C18 Stage Tip clean-up and analyzed in Orbitrap Fusion Tribrid mass spectrometer interfaced with Proxeon Easy-nano LC II system (Thermo Scientific, Bremen, Germany). We identified a total of 6773 peptides with 28426 peptide spectral matches, which belonged to 1081 proteins. Gene ontology analysis has been carried out to determine the biological processes, molecular functions and cellular components associated with sperm protein. The biological process chiefly represented our data is an oxidation-reduction process (5%), spermatogenesis (2.5%) and spermatid development (1.4%). The highlighted molecular functions are ATP, and GTP binding (14%) and the prominent cellular components most observed in our data were nuclear membrane (1.5%), acrosomal vesicle (1.4%), and motile cilium (1.3%). Seventeen percent of sperm proteins identified in this study were involved in metabolic pathways. To the best of our knowledge, this data represents the first total sperm proteome from indigenous cattle, Malnad Gidda. We believe that our preliminary findings could provide a strong base for the future understanding of bovine sperm proteomics.

Keywords: Bos indicus, Malnad Gidda, mass spectrometry, spermatozoa

Procedia PDF Downloads 175

Authors: Henry O. Chibudike, Olubamike A. Adeyoju, Bolanle O. Oluwole, Kayode O. Adebowale, Bamidele I. Olu-Owolabi, Chinedum E. Chibudike

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Studies on the Mineral Content of Solojo Flour and Protein Isolates from the two varieties (DAS and BS) of Nigeria cultivated solojo cowpeas were conducted to determine their nutritional value. These inorganic elements or minerals were classified into 3 categories: the ultra-trace minerals, which are the third category; the microelements, also known as the trace minerals, in the second category; while the first category is the macro elements, also known as major minerals. Some of the macro-elements are Ca, P, Na and Cl; the second category, micro-elements include iron, copper, cobalt, potassium, magnesium, iodine, zinc, manganese, molybdenum, F, Cr, Se and S. Results show that the proportion of Sodium (Na) which is ingested into the body in the form of NaCl through food intake maintenance of body pH and to retain water ranged from 728.97 to 253.37 ppm (72.90 to 25.34 mg/100 g); 715.24 to 235.45 ppm; 735.28 to 270.37 ppm; 726.59 to 264.35ppm, for FFDAS, FFBS, DAS and BS respectively with all values of the germinated samples all bellow the control. While FFDAS iron content ranged from 4.25 to 13.50 mg/100 g; FFBS ranged from 3.15 to 12.56 mg/100 g; DAS ranged from 3.81 to 12.90 mg/100g; BS ranged from 3.42 to 9.40 mg/100 g. The values of the germinated flours were all greater than the ungerminated flour. Iron helps to transport oxygen round the body and also helps in red blood cells building and to convert food into needed energy by the body. While Manganese an element that is needed in micro quantity but necessary to convert food into energy, is also crucial for healthy bone and cartilage creation. Results also show that zinc quantity increased as germination proceeded, and the values ranged from 38.80 ppm to 230.00 ppm (3.880 mg/100 g to 23.00 mg/100 g; 0.003880% to 0.0230%); 40.84 to 250.01 ppm; 32.85 to 93.41 ppm; 37.07 to 115.00 ppm, for FFDAS, FFBS, DAS and BS respectively. The Ca content improved significantly (p<0.05) with sprouting; the value extended from 250.56 ppm to 760.03 ppm (25.056 to 76.00 mg/100g or 0.0251 to 0.0760 %); 400.40 to 998.22 ppm; 116.87 to 195.69 ppm; 113.48 to 220.75 ppm, for FFDAS, FFBS, DAS and BS respectively. Zinc element although needed at the micro level in the body, is essential for a strong immune system to keep the body in good health. It is also crucial for the maintenance of a healthy sense of taste and odor, while Calcium is critical for strong bones and teeth, blood coagulation, and muscle tightening and relaxation. Magnesium is needed to build enzymes and antioxidants and also for healthy bones, while Potassium is needed to maintain water balance, muscle movement, and nerve impulses. It functions in conjunction with Na to regulate blood pressure.

Keywords: Solojo cowpea, underutilized legumes, protein isolates, BS, DAS, ungerminated

Procedia PDF Downloads 34

Authors: Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Susmita Goswami, Prem D. Sharma, Prabir K. Paul

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The concept of organic agriculture has been accepted as novelty in Indian society, but there is no data available on the human pathogens colonizing plant parts due to such practices. Also, the pattern and mechanism of their colonization need to be understood in order to devise possible strategies for their prevention. In the present study, human pathogenic bacteria were isolated from organically grown tomato plants and five of them were identified as Klebsiella pneumoniae, Enterobacter ludwigii, Serratia fonticola, Stenotrophomonas maltophilia and Chryseobacterium jejuense. Tomato plants were grown in controlled aseptic conditions with 25±1˚C, 70% humidity and 12 hour L/D photoperiod. Six weeks old plants were divided into 6 groups of 25 plants each and treated as follows: Group 1: K. pneumonia, Group 2: E. ludwigii, Group 3: S. fonticola, Group 4: S. maltophilia, Group 5: C. jejuense, Group 6: Sterile distilled water (control). The inoculums for all treatments were prepared by overnight growth with uniform concentration of 108 cells/ml. Leaf samples from above groups were collected at 0.5, 2, 4, 6 and 24 hours post inoculation for the colony forming unit counts (CFU/cm2 of leaf area) of individual pathogens using leaf impression method. These CFU counts were used for the in vivo colonization assay and adherence assay of individual pathogens. Also, resistance of these pathogens to at least 12 antibiotics was studied. Based on these findings S. fonticola was found to be most prominently colonizing the phylloplane of tomato and was further studied. Tomato plants grown in controlled aseptic conditions same as mentioned above were divided into 2 groups of 25 plants each and treated as follows: Group 1: S. fonticola, Group 2: Sterile distilled water (control). Leaf samples from above groups were collected at 0, 24, 48, 72 and 96 hours post inoculation and homogenized in suitable buffers for surface and cell wall protein isolation. Protein samples thus obtained were subjected to isocratic SDS-gel electrophoresis and analyzed. It was observed that presence of S. fonticola could induce the expression of at least 3 additional cell wall proteins at different time intervals. Surface proteins also showed variation in the expression pattern at different sampling intervals. Further identification of these proteins by MALDI-MS and bioinformatics tools revealed the gene(s) involved in the interaction of S. fonticola with tomato phylloplane.

Keywords: cell wall proteins, human pathogenic bacteria, phylloplane, solanum lycopersicum

Procedia PDF Downloads 206

Authors: Gabriela Kalcikova, Gregor Marolt, Anita Jemec Kokalj, Andreja Zgajnar Gotvajn

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Since the 90’s, nanotechnology is one of the fastest growing fields of science. Nanomaterials are increasingly becoming part of many products and technologies. Metal oxide nanoparticles are among the most used nanomaterials. Zinc oxide nanoparticles (nZnO) is widely used due to its versatile properties; it has been used in products including plastics, paints, food, batteries, solar cells and cosmetic products. It is also a very effective photocatalyst used for water treatment. Such expanding application of nZnO increases their possible occurrence in the environment. In the aquatic ecosystem nZnO interact with natural environmental factors such as UV radiation, and thus it is essential to evaluate possible interaction between them. In this context, the aim of our study was to evaluate combined ecotoxicity of nZnO and Zn²⁺ on duckweed Lemna minor in presence or absence UV. Inhibition of vegetative growth of duckweed Lemna minor was monitored over a period of 7 days in multi-well plates. After the experiment, specific growth rate was determined. ZnO nanoparticles used were of primary size 13.6 ± 1.7 nm. The test was conducted with nominal nZnO and Zn²⁺ (in form of ZnCl₂) concentrations of 1, 10, 100 mg/L. Experiment was repeated with presence of natural intensity of UV (8h UV, 10 W/m² UVA, 0.5 W/m² UVB). Concentration of Zn during the test was determined by ICP-MS. In the regular experiment (absence of UV) the specific growth rate was slightly increased by low concentrations of nZnO and Zn²⁺ in comparison to control. However, 10 and 100 mg/L of Zn²⁺ resulted in 45% and 68% inhibition of the specific growth rate, respectively. In case of nZnO both concentrations (10 and 100 mg/L) resulted in similar ~ 30% inhibition and the response was not dose-dependent. The lack of the dose-response relationship is often observed in case of nanoparticles. The possible explanation is that the physical impact prevails instead of chemical ones. In the presence of UV the toxicity of Zn²⁺ was increased and 100 mg/L of Zn²⁺ caused total inhibition of the specific growth rate (100%). On the other hand, 100 mg/L of nZnO resulted in low inhibition (19%) in comparison to the experiment without UV (30%). It is thus expected, that tested nZnO is low photoactive, but could have a good UV absorption and/or reflective properties and thus protect duckweed against UV impacts. Measured concentration of Zn in the test suspension decreased only about 4% after 168h in the case of ZnCl₂. On the other hand concentration of Zn in nZnO test decreased by 80%. It is expected that nZnO were partially dissolved in the medium and at the same time agglomeration and sedimentation of particles took place and thus the concentration of Zn at the water level decreased. Results of our study indicated, that nZnO combined with UV of natural intensity does not increase toxicity of nZnO, but slightly protect the plant against UV negative effects. When Zn²⁺ and ZnO results are compared it seems that dissolved Zn plays a central role in the nZnO toxicity.

Keywords: duckweed, environmental factors, nanoparticles, toxicity

Procedia PDF Downloads 306

Authors: I. Bochev, A. Shterev, S. Kyurkchiev

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One of the factors associated with poor success rates in human in vitro fertilization (IVF) is the suboptimal culture conditions in which fertilization and early embryonic growth occur. Co-culture systems with helper cell lines appear to enhance the in vitro conditions and allow embryos to demonstrate improved in vitro development. The co-culture of human embryos with monolayers of autologous endometrial stromal cell (EnSCs) results in increased blastocyst development with a larger number of blastomeres, lower incidence of fragmentation and higher pregnancy rates in patients with recurrent implantation failure (RIF). The aim of the study was to examine the influence of autologous endometrial stromal cell (EnSC) co-culture on day 3 embryo quality by comparing the morphological status of the embryos from the same patients undergoing consecutive IVF/Intracytoplasmic sperm injection (ICSI) cycles without and with EnSC co-culture. This retrospective randomized study (2015-2017) includes 20 couples and a total of 46 IVF/ICSI cycles. Each patient couple included had at least two IVF/ICSI procedures – one with and one without autologous EnSC co-culture. Embryo quality was assessed at 68±1 hours in culture, according to Istanbul consensus criteria (2010). Day 3 embryos were classified into three groups: good – grade 1; fair – grade 2; poor – grade 3. Embryos from all cycles were divided into two groups (A – co-cultivated; B – not co-cultivated) and analyzed. Second, for each patient couple, embryos from matched IVF/ICSI cycles (with and without co-culture) were analyzed separately. When an analysis of co-cultivated day 3 embryos from all cycles was performed (n=137; group A), 43.1% of the embryos were graded as “good”, which was not significantly different from the respective embryo quality rate of 42.2% (p = NS) in group B (n=147) with non-co-cultivated embryos. The proportions of fair and poor quality embryos in group A and group B were similar as well – 11.7% vs 10.2% and 45.2% vs 47.6% (p=NS), respectively. Nevertheless, the separate embryo analysis by matched cycles for each couple revealed that in 65% of the cases the proportion of morphologically better embryos was increased in cycles with co-culture in comparison with those without co-culture. A decrease in this proportion after endometrial stromal cell co-cultivation was found in 30% of the cases, whereas no difference was observed in only one couple. The results demonstrated that there is no marked difference in the overall morphological quality between co-cultured and non-co-cultured embryos on day 3. However, in significantly greater percentage of couples the process of autologous EnSC co-culture could increase the proportion of morphologically improved day 3 embryos. By mimicking the in vivo relationship between embryo and maternal environment, co-culture in autologous EnSC system represents a perspective approach to improve the quality of embryos in cases with elevated risk for development of embryos with impaired morphology.

Keywords: autologous endometrial stromal cells, co-culture, day 3 embryo, morphological quality

Procedia PDF Downloads 206

Authors: Gurleen Kour, Mowkshi Khullar, B. K. Chandan, Parvinder Pal Singh, Kushalava Reddy Yumpalla, Gurunadham Munagala, Ram A. Vishwakarma, Zabeer Ahmed

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New chemotherapeutic compounds against multidrug-resistant Mycobacterium tuberculosis (Mtb) are urgently needed to combat drug resistance in tuberculosis (TB). Apart from in-vitro potency against the target, physiochemical properties and pharmacokinetic properties play an imperative role in the process of drug discovery. We have identified novel nitroimidazole derivatives with potential activity against mycobacterium tuberculosis. One lead candidates, MCD-017, which showed potent activity against H37Rv strain (MIC=0.5µg/ml) and was further evaluated in the process of drug development. Methods: Basic physicochemical parameters like solubility and lipophilicity (LogP) were evaluated. Thermodynamic solubility was determined in PBS buffer (pH 7.4) using LC/MS-MS. The partition coefficient (Log P) of the compound was determined between octanol and phosphate buffered saline (PBS at pH 7.4) at 25°C by the microscale shake flask method. The compound followed Lipinski’s rule of five, which is predictive of good oral bioavailability and was further evaluated for metabolic stability. In-vitro metabolic stability was determined in rat liver microsomes. The hepatotoxicity of the compound was also determined in HepG2 cell line. In vivo pharmacokinetic profile of the compound after oral dosing was also obtained using balb/c mice. Results: The compound exhibited favorable solubility and lipophilicity. The physical and chemical properties of the compound were made use of as the first determination of drug-like properties. The compound obeyed Lipinski’s rule of five, with molecular weight < 500, number of hydrogen bond donors (HBD) < 5 and number of hydrogen bond acceptors(HBA) not more then 10. The log P of the compound was less than 5 and therefore the compound is predictive of exhibiting good absorption and permeation. Pooled rat liver microsomes were prepared from rat liver homogenate for measuring the metabolic stability. 99% of the compound was not metabolized and remained intact. The compound did not exhibit cytoxicity in hepG2 cells upto 40 µg/ml. The compound revealed good pharmacokinetic profile at a dose of 5mg/kg administered orally with a half life (t1/2) of 1.15 hours, Cmax of 642ng/ml, clearance of 4.84 ml/min/kg and a volume of distribution of 8.05 l/kg. Conclusion : The emergence of multi drug resistance (MDR) and extensively drug resistant (XDR) Tuberculosis emphasize the requirement of novel drugs active against tuberculosis. Thus, the need to evaluate physicochemical and pharmacokinetic properties in the early stages of drug discovery is required to reduce the attrition associated with poor drug exposure. In summary, it can be concluded that MCD-017 may be considered a good candidate for further preclinical and clinical evaluations.

Keywords: mycobacterium tuberculosis, pharmacokinetics, physicochemical properties, hepatotoxicity

Procedia PDF Downloads 437

Authors: Ursula Das, Manavjit Singh Dhindsa, Kshirasagar Naik, Marzia Zaman, Richard Purcell, Srinivas Sampalli, Abdul Mutakabbir, Chung-Horng Lung, Thambirajah Ravichandran

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Wildland fires, also known as forest fires or wildfires, are exhibiting an alarming surge in frequency in recent times, further adding to its perennial global concern. Forest fires often lead to devastating consequences ranging from loss of healthy forest foliage and wildlife to substantial economic losses and the tragic loss of human lives. Despite the existence of substantial literature on the detection of active forest fires, numerous potential research avenues in forest fire management, such as preventative measures and ancillary effects of forest fires, remain largely underexplored. This paper undertakes a systematic review of these underexplored areas in forest fire research, meticulously categorizing them into distinct phases, namely pre-fire, during-fire, and post-fire stages. The pre-fire phase encompasses the assessment of fire risk, analysis of fuel properties, and other activities aimed at preventing or reducing the risk of forest fires. The during-fire phase includes activities aimed at reducing the impact of active forest fires, such as the detection and localization of active fires, optimization of wildfire suppression methods, and prediction of the behavior of active fires. The post-fire phase involves analyzing the impact of forest fires on various aspects, such as the extent of damage in forest areas, post-fire regeneration of forests, impact on wildlife, economic losses, and health impacts from byproducts produced during burning. A comprehensive understanding of the three stages is imperative for effective forest fire management and mitigation of the impact of forest fires on both ecological systems and human well-being. Artificial intelligence and machine learning (AI/ML) methods have garnered much attention in the cyber-physical systems domain in recent times leading to their adoption in decision-making in diverse applications including disaster management. This paper explores the current state of AI/ML applications for managing the activities in the aforementioned phases of forest fire. While conventional machine learning and deep learning methods have been extensively explored for the prevention, detection, and management of forest fires, a systematic classification of these methods into distinct AI research domains is conspicuously absent. This paper gives a comprehensive overview of the state of forest fire research across more recent and prominent AI/ML disciplines, including big data, classical machine learning, computer vision, explainable AI, generative AI, natural language processing, optimization algorithms, and time series forecasting. By providing a detailed overview of the potential areas of research and identifying the diverse ways AI/ML can be employed in forest fire research, this paper aims to serve as a roadmap for future investigations in this domain.

Keywords: artificial intelligence, computer vision, deep learning, during-fire activities, forest fire management, machine learning, pre-fire activities, post-fire activities

Procedia PDF Downloads 43

Authors: Likith Reddy Pinninti, Fredrik Ribsskog Staven, Leslie Robert Noble, Jorge Manuel de Oliveira Fernandes, Deepti Manjari Patel, Torstein Kristensen

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Understanding fish behavior is essential to improve animal welfare in aquaculture research. Behavioral traits can have a strong influence on fish health and habituation. To identify the genes and biological pathways responsible for lumpfish behavior, we performed an experiment to understand the interspecies relationship (mutualism) between the lumpfish and salmon. Also, we tested the correlation between the gene expression data vs. observational/physiological data to know the essential genes that trigger stress and swimming behavior in lumpfish. After the de novo assembly of the brain transcriptome, all the samples were individually mapped to the available lumpfish (Cyclopterus lumpus L.) primary genome assembly (fCycLum1.pri, GCF_009769545.1). Out of ~16749 genes expressed in brain samples, we found 267 genes to be statistically significant (P > 0.05) found only in odor and control (1), model and control (41) and salmon and control (225) groups. However, genes with |LogFC| ≥0.5 were found to be only eight; these are considered as differentially expressed genes (DEG’s). Though, we are unable to find the differential genes related to the behavioral traits from RNA-Seq data analysis. From the correlation analysis, between the gene expression data vs. observational/physiological data (serotonin (5HT), dopamine (DA), 3,4-Dihydroxyphenylacetic acid (DOPAC), 5-hydroxy indole acetic acid (5-HIAA), Noradrenaline (NORAD)). We found 2495 genes found to be significant (P > 0.05) and among these, 1587 genes are positively correlated with the Noradrenaline (NORAD) hormone group. This suggests that Noradrenaline is triggering the change in pigmentation and skin color in lumpfish. Genes related to behavioral traits like rhythmic, locomotory, feeding, visual, pigmentation, stress, response to other organisms, taxis, dopamine synthesis and other neurotransmitter synthesis-related genes were obtained from the correlation analysis. In KEGG pathway enrichment analysis, we find important pathways, like the calcium signaling pathway and adrenergic signaling in cardiomyocytes, both involved in cell signaling, behavior, emotion, and stress. Calcium is an essential signaling molecule in the brain cells; it could affect the behavior of fish. Our results suggest that changes in calcium homeostasis and adrenergic receptor binding activity lead to changes in fish behavior during stress.

Keywords: behavior, De novo, lumpfish, salmon

Procedia PDF Downloads 152

Authors: Simrat Kaur, Fatema Diwan, Brad Reddersen

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The unsustainable and injudicious use of natural renewable resources beyond the self-replenishment limits of our planet has proved catastrophic. Most of the Earth’s resources, including land, water, minerals, and biodiversity, have been overexploited. Owing to this, there is a steep rise in the global events of natural calamities of contrasting nature, such as torrential rains, storms, heat waves, rising sea levels, and megadroughts. These are all interconnected through common elements, namely oceanic currents and land’s the green cover. The deforestation fueled by the ‘economic elites’ or the global players have already cleared massive forests and ecological biomes in every region of the globe, including the Amazon. These were the natural carbon sinks prevailing and performing CO2 sequestration for millions of years. The forest biomes have been turned into mono cultivation farms to produce feedstock crops such as soybean, maize, and sugarcane; which are one of the biggest green house gas emitters. Such unsustainable agriculture practices only provide feedstock for livestock and food processing industries with huge carbon and water footprints. These are two main factors that have ‘cause and effect’ relationships in the context of climate change. In contrast to organic and sustainable farming, the mono-cultivation practices to produce food, fuel, and feedstock using chemicals devoid of the soil of its fertility, abstract surface, and ground waters beyond the limits of replenishment, emit green house gases, and destroy biodiversity. There are numerous cases across the planet where due to overuse; the levels of surface water reservoir such as the Lake Mead in Southwestern USA and ground water such as in Punjab, India, have deeply shrunk. Unlike the rain fed food production system on which the poor communities of the world relies; the blue water (surface and ground water) dependent mono-cropping for industrial and processed food create water deficit which put the burden on the domestic users. Excessive abstraction of both surface and ground waters for high water demanding feedstock (soybean, maize, sugarcane), cereal crops (wheat, rice), and cash crops (cotton) have a dual and synergistic impact on the global green house gas emissions and prevalence of megadroughts. Both these factors have elevated global temperatures, which caused cascading events such as soil water deficits, flash fires, and unprecedented burning of the woods, creating megafires in multiple continents, namely USA, South America, Europe, and Australia. Therefore, it is imperative to reduce the green and blue water footprints of agriculture and industrial sectors through recycling of black and gray waters. This paper explores various opportunities for successful implementation of wastewater management for drought preparedness in high risk communities.

Keywords: wastewater, drought, biodiversity, water footprint, nutrient recovery, algae

Procedia PDF Downloads 81

Authors: Brad Stappenbelt

Abstract:

The phenomenon of vortex-induced vibration (VIV) for elastically restrained cylindrical structures in cross-flows is relatively well investigated. The utility of this mechanism in harvesting energy from marine current and tidal flows is however arguably still in its infancy. With relatively few moving components, a flow-induced vibration-based energy conversion device augers low complexity compared to the commonly employed turbine design. Despite the interest in this concept, a practical device has yet to emerge. It is desirable for optimal system performance to design for a very low mass or mass moment of inertia ratio. The device operating range, in particular, is maximized below the vortex-induced vibration critical point where an infinite resonant response region is realized. An unfortunate consequence of this requirement is large buoyancy forces that need to be mitigated by gravity-based, suction-caisson or anchor mooring systems. The focus of this paper is the testing of a novel VIV marine current energy harvesting configuration that utilizes a symmetrical and balanced pair of horizontal pivoted cylinders. The results of several years of experimental investigation, utilizing the University of Wollongong fluid mechanics laboratory towing tank, are analyzed and presented. A reduced velocity test range of 0 to 60 was covered across a large array of device configurations. In particular, power take-off damping ratios spanning from 0.044 to critical damping were examined in order to determine the optimal conditions and hence the maximum device energy conversion efficiency. The experiments conducted revealed acceptable energy conversion efficiencies of around 16% and desirable low flow-speed operating ranges when compared to traditional turbine technology. The potentially out-of-phase spanwise VIV cells on each arm of the device synchronized naturally as no decrease in amplitude response and comparable energy conversion efficiencies to the single cylinder arrangement were observed. In addition to the spatial design benefits related to the horizontal device orientation, the main advantage demonstrated by the current symmetrical horizontal configuration is to allow large velocity range resonant response conditions without the excessive buoyancy. The novel configuration proposed shows clear promise in overcoming many of the practical implementation issues related to flow-induced vibration marine current energy harvesting.

Keywords: flow-induced vibration, vortex-induced vibration, energy harvesting, tidal energy

Procedia PDF Downloads 129

Authors: Agumah N. B. Orji, M. U., Oru C. M., Ugbo, E. N., Onwuliri E. A Nwakaeze, E. A.,

Abstract:

ERG11 gene has been reported to be one of the genes whose expression is responsible for resistance of Candida to various triazole drugs, which are first line treatment for candidiasis. This study was carried out to determine the prevalence of Triazole (Fluconazole and voriconazole) resistant Candida albicans expressing ERG11 gene from adult females in Abakaliki. Urine and vaginal swab samples were randomly collected from volunteers after obtaining their consent to participate in the study. A total of 565 adult females participated in the study. A total of 340 urine specimens and 288 vaginal swab specimens were collected. Direct wet mount technique, as well as culture in Trichomonas broth, were used to examine the urine and vaginal swab specimens for the presence of motile Trichomonads. The Trichomonas broth used was selective for both T. vaginalis and C. albicans. Broths that yielded budding yeast cells after microscopy were subcultured on to Sabouraud dextrose agar, after which Germ tube test was carried out to confirm the presence of C. albicans. Biochemical tests, including carbohydrate fermentation and urease utilization, were also performed. Antibiogram of C. albicans isolates obtained from this study was carried out using commercially available azole drugs. Fluconazole and voriconazole were selected as Triazole drugs used for this study. Nystatin was used as a tangential control. An MIC test was carried out with E-strips on some of the resistant C. albicans isolates A total of 6 isolates that resisted all the azole drugs were selected and screened for the presence of ERG11 gene using Reverse transcriptase polymerase chain reaction technique. The total prevalence recorded for C. albicans was 13.0%. Frequency was statistically higher in Pregnant (7.96%) than non pregnant (5.09%) volunteers (X2=0.94 at P=0.05). With respect to clinical samples, frequency was higher in vaginal swabs samples (7.96%) than Urine samples (5.09%) (X2=9.05 at P=0.05). Volunteers within the age group 26-30 years recorded the highest prevalence (4.46%), while those within the age group 36-40 years recorded the lowest at 1.27%(X2=4.34 at P=0.05). In pregnant female participants, the highest frequency was recorded with those in their 3rd trimester (4.14%), while lowest incidence was recorded for those in their first trimester (0.80%). Antibiogram results from this study showed that C. albicans isolates obtained from this study resisted Fluconazole (72%) more than Voriconazole (57%). Only one out of the six selected isolates yielded resistance in the MIC test. Results obtained from the RT-PCR showed that there was no expression of ERG11 gene among the fluconazole resistant isolates of C. albicans. Observed resistance may be due to other factors other than expression of ERG11 gene.

Keywords: candida, ERG11, triazole, nigeria

Procedia PDF Downloads 130

Authors: Kimberly Samaha

Abstract:

In order to attract new types of investors into the emerging Bio-Economy, new methodologies to analyze investment risk are needed. The Bio-Hub Ecosystem model was developed to address a critical area of concern within the global energy market regarding the use of biomass as a feedstock for power plants. This study looked at repurposing existing biomass-energy plants into Circular Zero-Waste Bio-Hub Ecosystems. A Bio-Hub model that first targets a ‘whole-tree’ approach and then looks at the circular economics of co-hosting diverse industries (wood processing, aquaculture, agriculture) in the vicinity of the Biomass Power Plants facilities. This study modeled the economics and risk strategies of cradle-to-cradle linkages to incorporate the value-chain effects on capital/operational expenditures and investment risk reductions using a proprietary techno-economic model that incorporates investment risk scenarios utilizing the Monte Carlo methodology. The study calculated the sequential increases in profitability for each additional co-host on an operating forestry-based biomass energy plant in West Enfield, Maine. Phase I starts with the base-line of forestry biomass to electricity only and was built up in stages to include co-hosts of a greenhouse and a land-based shrimp farm. Phase I incorporates CO2 and heat waste streams from the operating power plant in an analysis of lowering and stabilizing the operating costs of the agriculture and aquaculture co-hosts. Phase II analysis incorporated a jet-fuel biorefinery and its secondary slip-stream of biochar which would be developed into two additional bio-products: 1) A soil amendment compost for agriculture and 2) A biochar effluent filter for the aquaculture. The second part of the study applied the Monte Carlo risk methodology to illustrate how co-location derisks investment in an integrated Bio-Hub versus individual investments in stand-alone projects of energy, agriculture or aquaculture. The analyzed scenarios compared reductions in both Capital and Operating Expenditures, which stabilizes profits and reduces the investment risk associated with projects in energy, agriculture, and aquaculture. The major findings of this techno-economic modeling using the Monte Carlo technique resulted in the masterplan for the first Bio-Hub to be built in West Enfield, Maine. In 2018, the site was designated as an economic opportunity zone as part of a Federal Program, which allows for Capital Gains tax benefits for investments on the site. Bioenergy facilities are currently at a critical juncture where they have an opportunity to be repurposed into efficient, profitable and socially responsible investments, or be idled and scrapped. The Bio-hub Ecosystems techno-economic analysis model is a critical model to expedite new standards for investments in circular zero-waste projects. Profitable projects will expedite adoption and advance the critical transition from the current ‘take-make-dispose’ paradigm inherent in the energy, forestry and food industries to a more sustainable Bio-Economy paradigm that supports local and rural communities.

Keywords: bio-economy, investment risk, circular design, economic modelling

Procedia PDF Downloads 88

Authors: Iuri Salukvadze

Abstract:

Construction takes importance role in the history of mankind, there is not a single thing-product in our lives in which the builder’s work was not to be materialized, because to create all of it requires setting up factories, roads, and bridges, etc. The function of the Republic of Georgia, as part of the connecting Europe-Asia transport corridor, is significantly increased. In the context of transit function a large part of the cargo traffic belongs to motor transport, hence the improvement of motor roads transport infrastructure is rather important and rise the new, increased operational demands for existing as well as new motor roads. Construction of the durable road surface is related to rather large values, but because of high transport-operational properties, such as high-speed, less fuel consumption, less depreciation of tires, etc. If the traffic intensity is high, therefore the reimbursement of expenses occurs rapidly and accordingly is increasing income. If the traffic intensity is relatively small, it is recommended to use lightened structures of road carpet in order to pay for capital investments amounted to no more than normative one. The road carpet is divided into the following basic types: asphaltic concrete and cement concrete. Asphaltic concrete is the most perfect type of road carpet. It is arranged in two or three layers on rigid foundation and will be compacted. Asphaltic concrete is artificial building material, which due stratum will be selected and measured from stone skeleton and sand, interconnected by bitumen and a mixture of mineral powder. Less strictly selected similar material is called as bitumen-mineral mixture. Asphaltic concrete is non-rigid building material and well durable on vertical loadings; it is less resistant to the impact of horizontal forces. The cement concrete is monolithic and durable material, it is well durable the horizontal loads and is less resistant related to vertical loads. The cement concrete consists from strictly selected, measured stone material and sand, the binder is cement. The cement concrete road carpet represents separate slabs of sizes from 3 ÷ 5 op to 6 ÷ 8 meters. The slabs are reinforced by a rather complex system. Between the slabs are arranged seams that are designed for avoiding of additional stresses due temperature fluctuations on the length of slabs. For the joint behavior of separate slabs, they are connected by metal rods. Rods provide the changes in the length of slabs and distribute to the slab vertical forces and bending moments. The foundation layers will be extremely durable, for that is required high-quality stone material, cement, and metal. The qualification work aims to: in order for improvement of traffic conditions on motor roads to prolong operational conditions and improving their characteristics. The work consists from three chapters, 80 pages, 5 tables and 5 figures. In the work are stated general concepts as well as carried out by various companies using modern methods tests and their results. In the chapter III are stated carried by us tests related to this issue and specific examples to improving the operational characteristics.

Keywords: asphalt, cement, cylindrikal sample of asphalt, building

Procedia PDF Downloads 200

Authors: Ying Li, Rui Hu, Shizhen Chen, Xin Zhou, Yunhuang Yang

Abstract:

Prostate cancer is one of the leading causes of cancer-related death among men. Prostate-specific antigen (PSA), a specific product of prostatic epithelial cells, is an important indicator of prostate cancer. Though PSA is not a specific serum biomarker for the screening of prostate cancer, it is recognized as an indicator for prostate cancer recurrence and response to therapy for patient’s post-prostatectomy. Since radical prostatectomy eliminates the source of PSA production, serum PSA levels fall below 50 pg/mL, and may be below the detection limit of clinical immunoassays (current clinical immunoassay lower limit of detection is around 10 pg/mL). Many clinical studies have shown that intervention at low PSA levels was able to improve patient outcomes significantly. Therefore, ultra-sensitive and precise assays that can accurately quantify extremely low levels of PSA (below 1-10 pg/mL) will facilitate the assessment of patients for the possibility of early adjuvant or salvage treatment. Currently, the commercially available ultra-sensitive ELISA kit (not used clinically) can only reach a detection limit of 3-10 pg/mL. Other platforms developed by different research groups could achieve a detection limit as low as 0.33 pg/mL, but they relied on sophisticated instruments to get the final readout. Herein we report a microfluidic platform for point-of-care (POC) detection of PSA with a detection limit of 0.5 pg/mL and without the assistance of any equipment. This platform is based on a previously reported volumetric-bar-chart chip (V-Chip), which applies platinum nanoparticles (PtNPs) as the ELISA probe to convert the biomarker concentration to the volume of oxygen gas that further pushes the red ink to form a visualized bar-chart. The length of each bar is used to quantify the biomarker concentration of each sample. We devised a long reading channel V-Chip (LV-Chip) in this work to achieve a wide detection window. In addition, LV-Chip employed a unique enzyme-free ELISA probe that enriched PtNPs significantly and owned 500-fold enhanced catalytic ability over that of previous V-Chip, resulting in a significantly improved detection limit. LV-Chip is able to complete a PSA assay for five samples in 20 min. The device was applied to detect PSA in 50 patient serum samples, and the on-chip results demonstrated good correlation with conventional immunoassay. In addition, the PSA levels in finger-prick whole blood samples from healthy volunteers were successfully measured on the device. This completely stand-alone LV-Chip platform enables convenient POC testing for patient follow-up in the physician’s office and is also useful in resource-constrained settings.

Keywords: point-of-care detection, microfluidics, PSA, ultra-sensitive

Procedia PDF Downloads 91