Search results for: broth micro dilution assay

Authors: Ouahiba Taamallah, Tarek Litim

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The study deals with the burnishing effect of AISI 52100 steel and parameters influence (Py, i and f on surface integrity. The results show that the optimal effects are closely related to the treatment parameters. With a 92% improvement in roughness, SB can be defined as a finishing operation within the machining range. Due to 85% gain in consolidation rate, this treatment constitutes an efficient process for work-hardening of material. In addition, a statistical study based on regression and Taguchi's design has made it possible to develop mathematical models to predict output responses according to the studied burnishing parameters. Response Surface Methodology RSM showed a simultaneous influence of the burnishing parameters and to observe the optimal parameters of the treatment. ANOVA Analysis of results led to validate the prediction model with a determination coefficient R2=94.60% and R2=93.41% for surface roughness and micro-hardness, respectively. Furthermore, a multi-objective optimization allowed to identify a regime characterized by P=20 Kgf, i=5 passes and f=0.08 mm.rev-1, which favors minimum surface roughness and a maximum of micro-hardness. The result was validated by a composite desirability D_i=1 for both surface roughness and microhardness, respectively. Applying optimal parameters, burnishing showed its beneficial effects in fatigue resistance, especially for imposed loading in the low cycle fatigue of the material where the lifespan increased by 90%.

Keywords: AISI 52100 steel, burnishing, Taguchi, fatigue

Procedia PDF Downloads 178

Authors: Syed A. Bukhari, Ankur Goswmai, Dale Hume, Thomas Thundat

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Here we demonstrate mechanical detection of photo-induced Insulator to metal transition (MIT) in ultra-thin vanadium dioxide (VO₂) micro strings by using < 100 µW of optical power. Highly focused laser beam heated the string locally resulting in through plane and along axial heat diffusion. Localized temperature increase can cause temperature rise > 60 ºC. The heated region of VO₂ can transform from insulating (monoclinic) to conducting (rutile) phase leading to lattice compressions and stiffness increase in the resonator. The mechanical frequency of the resonator can be tuned by changing optical power and wavelength. The first mode resonance frequency was tuned in three different ways. A decrease in frequency below a critical optical power, a large increase between 50-120 µW followed by a large decrease in frequency for optical powers greater than 120 µW. The dynamic mechanical response was studied as a function of incident optical power and gas pressure. The resonance frequency and amplitude of vibration were found to be decreased with increasing laser power from 25-38 µW and increased by1-2 % when the laser power was further increased to 52 µW. The transition in films was induced and detected by a single pump and probe source and by employing external optical sources of different wavelengths. This trend in dynamic parameters of the strings can be co-related with reversible Insulator to metal transition in VO₂ films which creates change in density of the material and hence the overall stiffness of the strings leading to changes in string dynamics. The increase in frequency at a particular optical power manifests a transition to a more ordered metallic phase which tensile stress onto the string. The decrease in frequency at higher optical powers can be correlated with poor phonon thermal conductivity of VO₂ in conducting phase. Poor thermal conductivity of VO₂ can force in-plane penetration of heat causing the underneath SiN supporting VO₂ which can result as a decrease in resonance frequency. This noninvasive, non-contact laser-based excitation and detection of Insulator to metal transition using micro strings resonators at room temperature and with laser power in few µWs is important for low power electronics, and optical switching applications.

Keywords: thermal conductivity, vanadium dioxide, MEMS, frequency tuning

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Authors: Jose Neri, Fabrice Canto, Alastair Magnaldo, Laurent Guillerme, Vincent Dugas

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A miniaturized and automated approach for the volumetric titration of free nitric acid in U(VI) solutions is presented. Free acidity measurement refers to the acidity quantification in solutions containing hydrolysable heavy metal ions such as U(VI), U(IV) or Pu(IV) without taking into account the acidity contribution from the hydrolysis of such metal ions. It is, in fact, an operation having an essential role for the control of the nuclear fuel recycling process. The main objective behind the technical optimization of the actual ‘beaker’ method was to reduce the amount of radioactive substance to be handled by the laboratory personnel, to ease the instrumentation adjustability within a glove-box environment and to allow a high-throughput analysis for conducting more cost-effective operations. The measurement technique is based on the concept of the Taylor-Aris dispersion in order to create inside of a 200 μm x 5cm circular cylindrical micro-channel a linear concentration gradient in less than a second. The proposed analytical methodology relies on the actinide complexation using pH 5.6 sodium oxalate solution and subsequent alkalimetric titration of nitric acid with sodium hydroxide. The titration process is followed with a CCD camera for fluorescence detection; the neutralization boundary can be visualized in a detection range of 500nm- 600nm thanks to the addition of a pH sensitive fluorophore. The operating principle of the developed device allows the active generation of linear concentration gradients using a single cylindrical micro channel. This feature simplifies the fabrication and ease of use of the micro device, as it does not need a complex micro channel network or passive mixers to generate the chemical gradient. Moreover, since the linear gradient is determined by the liquid reagents input pressure, its generation can be fully achieved in faster intervals than one second, being a more timely-efficient gradient generation process compared to other source-sink passive diffusion devices. The resulting linear gradient generator device was therefore adapted to perform for the first time, a volumetric titration on a chip where the amount of reagents used is fixed to the total volume of the micro channel, avoiding an important waste generation like in other flow-based titration techniques. The associated analytical method is automated and its linearity has been proven for the free acidity determination of U(VI) samples containing up to 0.5M of actinide ion and nitric acid in a concentration range of 0.5M to 3M. In addition to automation, the developed analytical methodology and technique greatly improves the standard off-line oxalate complexation and alkalimetric titration method by reducing a thousand fold the required sample volume, forty times the nuclear waste per analysis as well as the analysis time by eight-fold. The developed device represents, therefore, a great step towards an easy-to-handle nuclear-related application, which in the short term could be used to improve laboratory safety as much as to reduce the environmental impact of the radioanalytical chain.

Keywords: free acidity, lab-on-a-chip, linear concentration gradient, Taylor-Aris dispersion, volumetric titration

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Authors: Shanakr Bhattarai, V. S. Tiwari, Barak Akabayov

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The plummeting number of infections and death is due to the development of drug-resistant bacteria. In addition, the number of approved antibiotic drugs by the Food and Drug Administration (FDA) is insufficient. Therefore, developing new drugs and finding novel targets for central metabolic pathways in bacteria is urgently needed. One of the promising targets is DNA replication machinery which consists of many essential proteins and enzymes. DnaG primase is an essential enzyme and a central part of the DNA replication machinery. DnaG primase synthesizes short RNA primers that initiate the Okazaki fragments by the lagging strand DNA polymerase. Therefore, it is reasonable to assume that inhibition of primase activity will stall DNA replication and prevent bacterial proliferation. We did the expression and purification of eight different bacterial DnaGs (Mycobacterium tuberculosis(Mtb), Bacillus anthracis (Ba), Mycobacterium smegmatis (Msmeg), Francisella tularencis (Ft), Vibrio cholerae (Vc) and Yersinia pestis (Yp), Staphylococcus aureus(Saureus), Escherichia coli(Ecoli)) followed by the radioactive activity assay. After obtaining the pure and active protein DnaG, we synthesized the inhibitors for them. The inhibitors were divided into five different groups, each containing five molecules, and the cocktail inhibition assay was performed against each DnaGs. The groups of molecules inhibiting the DnaGs were further tested with individual molecules belonging to inhibiting groups. Each molecule showing inhibition was titrated against the corresponding DnaGs to find IC50. We got a molecule(VS167) that acted as broad inhibitors, inhibiting all eight DnaGs. Molecules VS180 and VS186 inhibited seven DnaGs (except Saureus). Similarly, two molecules(VS 173, VS176) inhibited five DnaGs (Mtb, Ba, Ft, Yp, Ecoli). VS261 inhibited four DnaGs (Mtb, Ba, Ft, Vc). MS50 inhibited Ba and Vc DnaGs. And some of the inhibitors inhibited only one DnaGs. Thus we found the broad and specific inhibitors for different bacterial DnaGs, and their Structure-activity analysis(SAR) was done. Further, We tried to explain the similarities among the enzyme DnaGs from different bacteria based on their inhibition pattern.

Keywords: DNA replication, DnaG, okazaki fragments, antibiotic drugs

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Authors: Igbakura I. Luga, Alex A. Adikwu

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A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21^st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus

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Authors: Sutapa Som Chaudhury, Chitrangada Das Mukhopadhyay

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Alzheimer’s disease is a well-known form of dementia since its discovery in 1906. Current Food and Drug Administration approved medications e.g. cholinesterase inhibitors, memantine offer modest symptomatic relief but do not play any role in disease modification or recovery. In last three decades many small molecules, chaperons, synthetic peptides, partial β-secretase enzyme blocker have been tested for the development of a drug against Alzheimer though did not pass the 3rd clinical phase trials. Here in this study, we designed a PEGylated, aminogenic, tripeptidic polymer with two different molecular weights based on the aggregation prone amino acid sequence 17-20 in amyloid beta (Aβ) 1-42. Being conjugated with poly-ethylene glycol (PEG) which self-assembles into hydrophilic nanoparticles, these PEGylated tripeptides constitute a very good drug delivery system crossing the blood brain barrier while the peptide remains protected from proteolytic degradation and non-specific protein interactions. Moreover, being completely aminogenic they would not raise any side effects. These peptide inhibitors were evaluated for their effectiveness against Aβ42 fibrillation at an early stage of oligomer to fibril formation as well as preformed fibril clearance via Thioflavin T (ThT) assay, dynamic light scattering analyses, atomic force microscopy and scanning electron microscopy. The inhibitors were proved to be safe at a higher concentration of 20µM by the reduction assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye. Moreover, SHSY5Y neuroblastoma cells have shown a greater survivability when treated with the inhibitors following Aβ42 fibril and oligomer treatment as compared with the control Aβ42 fibril and/or oligomer treated neuroblastoma cells. These make the peptidic inhibitors a promising compound in the aspect of the discovery of alternative medication for Alzheimer’s disease.

Keywords: Alzheimer’s disease, alternative medication, amyloid beta, PEGylated peptide

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Authors: Chloe Richards, Adrian Delgado Ollero, Yan Delaure, Fiona Regan

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Growing concern about the natural environment has accelerated the search for non-toxic, but at the same time, economically reasonable, antifouling materials. Bioinspired surfaces, due to their nano and micro topographical antifouling capabilities, provide a hopeful approach to the design of novel antifouling surfaces. Biological organisms are known to have highly evolved and complex topographies, demonstrating antifouling potential, i.e. shark skin. Previous studies have examined the antifouling ability of topographic patterns, textures and roughness scales found on natural organisms. One of the mechanisms used to explain the adhesion of cells to a substrate is called attachment point theory. Here, the fouling organism experiences increased attachment where there are multiple attachment points and reduced attachment, where the number of attachment points are decreased. In this study, an attempt to characterize the microtopography of the common brill fish, Scophthalmus rhombus, was undertaken. Scophthalmus rhombus is a small flatfish of the family Scophthalmidae, inhabiting regions from Norway to the Mediterranean and the Black Sea. They reside in shallow sandy and muddy coastal areas at depths of around 70 – 80 meters. Six engineered surfaces (inspired by the Brill fish scale) produced by a 2-photon polymerization (2PP) process were evaluated for their potential as an antifouling solution for incorporation onto tidal energy blades. The micro-textures were analyzed for their AF potential under both static and dynamic laboratory conditions using two laboratory grown diatom species, Amphora coffeaeformis and Nitzschia ovalis. The incorporation of a surface topography was observed to cause a disruption in the growth of A. coffeaeformis and N. ovalis cells on the surface in comparison to control surfaces. This work has demonstrated the importance of understanding cell-surface interaction, in particular, topography for the design of novel antifouling technology. The study concluded that biofouling can be controlled by physical modification, and has contributed significant knowledge to the use of a successful novel bioinspired AF technology, based on Brill, for the first time.

Keywords: attachment point theory, biofouling, Scophthalmus rhombus, topography

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Authors: Zongmin Ma, Shaowen Zhang, Yueping Fu, Jun Tang, Yunbo Shi, Jun Liu

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Solid-state quantum sensors are attracting wide interest because of their high sensitivity at room temperature. In particular, spin properties of nitrogen–vacancy (NV) color centres in diamond make them outstanding sensors of magnetic fields, electric fields and temperature under ambient conditions. Much of the work on NV magnetic sensing has been done so as to achieve the smallest volume, high sensitivity of NV ensemble-based magnetometry using micro-cavity, light-trapping diamond waveguide (LTDW), nano-cantilevers combined with MEMS (Micro-Electronic-Mechanical System) techniques. Recently, frequency-modulated microwaves with continuous optical excitation method have been proposed to achieve high sensitivity of 6 μT/√Hz using individual NV centres at nanoscale. In this research, we built-up an experiment to measure static magnetic field through continuous wave optical excitation with frequency-modulated microwaves method under continuous illumination with green pump light at 532 nm, and bulk diamond sample with a high density of NV centers (1 ppm). The output of the confocal microscopy was collected by an objective (NA = 0.7) and detected by a high sensitivity photodetector. We design uniform and efficient excitation of the micro strip antenna, which is coupled well with the spin ensembles at 2.87 GHz for zero-field splitting of the NV centers. Output of the PD signal was sent to an LIA (Lock-In Amplifier) modulated signal, generated by the microwave source by IQ mixer. The detected signal is received by the photodetector, and the reference signal enters the lock-in amplifier to realize the open-loop detection of the NV atomic magnetometer. We can plot ODMR spectra under continuous-wave (CW) microwave. Due to the high sensitivity of the lock-in amplifier, the minimum detectable value of the voltage can be measured, and the minimum detectable frequency can be made by the minimum and slope of the voltage. The magnetic field sensitivity can be derived from η = δB√T corresponds to a 10 nT minimum detectable shift in the magnetic field. Further, frequency analysis of the noise in the system indicates that at 10Hz the sensitivity less than 10 nT/√Hz.

Keywords: nitrogen-vacancy (NV) centers, frequency-modulated microwaves, magnetic field sensitivity, noise density

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Authors: Maria Lucia G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Isosporiasis is an affliction caused by coccidial protozoa belonging to genera Isospora spp. or Cystoisospora spp., which may parasitize the small and large intestines of dogs, of which neonates and young animals present higher risk of infection. This study aims at reporting a case of isosporiasis in neonate Pitbull dogs, as well as the diagnosis and treatment. Seven Pitbull puppies were admitted to the São Paulo State University (UNESP) Veterinary Hospital, Botucatu, São Paulo, Brazil, with history of yellowish diarrhea without mucus or blood for the past two days. The animals were five days old. The history of the mother, a primiparous two-year-old, revealed that she was properly vaccinated, not de-wormed and did not present diarrhea. The clinical examination revealed that the neonates weighted between 308 and 360 grams, and presented normal reflexes, moderate dehydration, body temperatures between 36.8 and 37.2 ºC, blood sugar between 103 and 124 mg/dL and normal appetite. A full blood count and a parasitology assay were performed to aid in the diagnosis. The full blood count detected eosinophilia, without any other relevant alterations. The parasitology assay (Willis-Molly & Faust) revealed the presence of Cystoisospora spp. The treatment was instituted with heated fluid therapy with Ringer’s Lactate (4 mL/100 g, subcutaneous) and antibiotic therapy with sulfamethoxazole-trimethoprim (15 mg/kg, orally) every 12 hours for ten days. The mother and other dogs that came in contact with the newborns were also treated. The environment was disinfected for 10 minutes with 1.6% quaternary ammonium. After 10 days, the newborns presented normal clinical signs and no alterations in the full blood count. Isosporiasis is an affliction with high mortality rates in litters that should be diagnosed and treated as soon as possible to increase the survival rates in these patients.

Keywords: Cystoisospora spp., neonatal infection, puppies, diarrhea,

Procedia PDF Downloads 118

Authors: Ryu Wonjin, Choi Hyeok, Park Joonhong

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Overlay welding for improving surface properties is a method of the surface treatments which improve surface properties of material by welding materials of alloy having corrosion resistance on the basic material surface. Overlay welding affects contents of chemical components and weld hardness from different parts by dilution of the lamination layer thickness, and it determines surface properties. Therefore, overlay welding has to take into account thickness of the lamination layers with the process. As a result in this study examined contents of Fe, weldability of the base metal and monel materials, hardness and surface flatness from different parts according to each the lamination layer parameters by overlay welding monel materials with corrosion resources to the base material of carbon steel. Through this, evaluated effect by the lamination layer parameters of welding and presented decision methods of the lamination layer parameters of the overlay welding by the purpose of use.

Keywords: clad pipe, lamination layer parameters, monel, overlay welding

Procedia PDF Downloads 261

Authors: Liang-Jen Wang, Sung-Chou Li, Yuan-Ming Yeh, Sheng-Yu Lee, Ho-Chang Kuo, Chia-Yu Yang

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Background: Dysbiosis in the gut microbial community might be involved in the pathophysiology of attention deficit/hyperactivity disorder (ADHD). The fungal component of the gut microbiome, namely the mycobiota, is a hyperdiverse group of multicellular eukaryotes that can influence host intestinal permeability. This study therefore aimed to investigate the impact of fungal mycobiome dysbiosis and intestinal permeability on ADHD. Methods: Faecal samples were collected from 35 children with ADHD and from 35 healthy controls. Total DNA was extracted from the faecal samples, and the internal transcribed spacer (ITS) regions were sequenced using high-throughput next-generation sequencing (NGS). The fungal taxonomic classification was analysed using bioinformatics tools, and the differentially expressed fungal species between the ADHD and healthy control groups were identified. An in vitro permeability assay (Caco-2 cell layer) was used to evaluate the biological effects of fungal dysbiosis on intestinal epithelial barrier function. Results: The β-diversity (the species diversity between two communities), but not α-diversity (the species diversity within a community), reflected the differences in fungal community composition between ADHD and control groups. At the phylum level, the ADHD group displayed a significantly higher abundance of Ascomycota and significantly lower abundance of Basidiomycota than the healthy control group. At the genus level, the abundance of Candida (especially Candida albicans) was significantly increased in ADHD patients compared to the healthy controls. In addition, the in vitro cell assay revealed that C. albicans secretions significantly enhanced the permeability of Caco-2 cells. Conclusions: The current study is the first to explore altered gut mycobiome dysbiosis using the NGS platform in ADHD. The findings from this study indicated that dysbiosis of the fungal mycobiome and intestinal permeability might be associated with susceptibility to ADHD.

Keywords: ADHD, fungus, gut–brain axis, biomarker, child psychiatry

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Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas

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Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.

Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid

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Authors: Lingling Shui, Shuting Xie

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As alternatives to electronic devices, optically active structures from responsive nanomaterials offer great opportunity buildup smart functional sensors. Hereby, we report on droplet microfluidics enabled construction and application of photonic microcapsules (PMCs) for colorimetric temperature microsensors, enabling miniaturization for injectable local micro-area sensing and integration for large-area sensing. Monodispersed PMCs are produced by in-situ photopolymerization of hydrogel shells of cholesteric liquid crystal (CLC)-in-water-in-oil double emulsion droplets prepared using microfluidic devices, with controllable physical structures and chemical compositions. Constructed PMCs exhibit thermal responsive structural color according to the selective Bragg reflection of CLC’s periodical helical structures within the microdroplet’s spherical confinement. Constructed PMCs with tunable size and composition have been successfully applied for monitoring the living cell extracellular temperature via co-incubation with cell suspension, and for detecting human body temperature via a flexible device from assembled PMCs. These PMCs could be flexibly applied in either micro-environment or large-area surface, enabling wide applications for precision temperature monitoring biological activities (e.g. cells or organs), optoelectronic devices working conditions (e.g. temperature indicators under extreme conditions), and etc.

Keywords: droplet, microfluidics, assembly, soft materials, microsensor

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Authors: M. R. Vijayakumar, Sanjay Kumar Singh, Lakshmi, Hithesh Dewangan, Sanjay Singh

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Trans resveratrol (RES) is a natural molecule proved for cancer preventive and therapeutic activities devoid of any potential side effects. However, the therapeutic application of RES in disease management is limited because of its rapid elimination from blood circulation thereby low biological half life in mammals. Therefore, the main objective of this study is to enhance the circulation as well as therapeutic efficacy using PEGylated liposomes. D-α-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS) is applied as steric surface decorating agent to prepare RES liposomes by thin film hydration method. The prepared nanoparticles were evaluated by various state of the art techniques such as dynamic light scattering (DLS) technique for particle size and zeta potential, TEM for shape, differential scanning calorimetry (DSC) for interaction analysis and XRD for crystalline changes of drug. Encapsulation efficiency and invitro drug release were determined by dialysis bag method. Cancer cell viability studies were performed by MTT assay, respectively. Pharmacokinetic studies were performed in sprague dawley rats. The prepared liposomes were found to be spherical in shape. Particle size and zeta potential of prepared formulations varied from 64.5±3.16 to 262.3±7.45 nm and -2.1 to 1.76 mV, respectively. DSC study revealed absence of potential interaction. XRD study revealed presence of amorphous form in liposomes. Entrapment efficiency was found to be 87.45±2.14 % and the drug release was found to be controlled up to 24 hours. Minimized MEC in MTT assay and tremendous enhancement in circulation time of RES PEGylated liposomes than its pristine form revealed that the stearic stabilized PEGylated liposomes can be an alternative tool to commercialize this molecule for chemopreventive and therapeutic applications in cancer.

Keywords: trans resveratrol, cancer nanotechnology, long circulating liposomes, bioavailability enhancement, liposomes for cancer therapy, PEGylated liposomes

Procedia PDF Downloads 577

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

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Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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Authors: Nika Botchorishvili, Olgha Giorgishvili

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Many-year investigations of the nature of damping creep of rigid bodies and materials led to the discovery of the fundamental character of this phenomenon. It occurs only when a rigid body comes in contact with a surface-active medium (liquid or gaseous), which brings about a decrease of the free surface energy of a rigid body as a result of adsorption, chemo-sorption or wetting. The reversibility of the process consists of a gradual disappearance of creep deformation when the action of a surface-active medium stops. To clarify the essence of processes, a physical model is constructed by using Griffith’s scheme and the well-known representation formulas of deformation origination and failure processes. The total creep deformation is caused by the formation and opening of microcracks throughout the material volume under the action of load. This supposedly happens in macroscopically homogeneous silicate and organic glasses, while in polycrystals (tuff, gypsum, steel) contacting with a surface-active medium micro crack are formed mainly on the grain boundaries. The creep of rubber is due to its swelling activated by stress. Acknowledgment: All experiments are financially supported by Shota Rustaveli National Science Foundation of Georgia. Study of Properties of Concretes (Both Ordinary and Compacted) Made of Local Building Materials and Containing Admixtures, and Their Further Introduction in Construction Operations and Road Building. DP2016_26. 22.12.2016.

Keywords: process reversibility, surface-active medium, Rebinder’s effect, micro crack, creep

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Authors: Kiurski S. Jelena, Aksentijević M. Snežana, Kecić S. Vesna, Oros B. Ivana

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The prosperity of electronic equipment in photocopying environment not only has improved work efficiency, but also has changed indoor air quality. Considering the number of photocopying employed, indoor air quality might be worse than in general office environments. Determining the contribution from any type of equipment to indoor air pollution is a complex matter. Non-methane hydrocarbons are known to have an important role of air quality due to their high reactivity. The presence of hazardous pollutants in indoor air has been detected in one photocopying shop in Novi Sad, Serbia. Air samples were collected and analyzed for five days, during 8-hr working time in three-time intervals, whereas three different sampling points were determined. Using multiple linear regression model and software package STATISTICA 10 the concentrations of occupational hazards and micro-climates parameters were mutually correlated. Based on the obtained multiple coefficients of determination (0.3751, 0.2389, and 0.1975), a weak positive correlation between the observed variables was determined. Small values of parameter F indicated that there was no statistically significant difference between the concentration levels of non-methane hydrocarbons and micro-climates parameters. The results showed that variable could be presented by the general regression model: y = b0 + b1xi1+ b2xi2. Obtained regression equations allow to measure the quantitative agreement between the variation of variables and thus obtain more accurate knowledge of their mutual relations.

Keywords: non-methane hydrocarbons, photocopying process, multiple regression analysis, indoor air quality, pollutant emission

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Authors: Mi Yuan

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Employment issues have become increasingly severe in contemporary society, with job seekers' criteria for work environments becoming more complex. However, most studies lack an analysis from the perspective of employment spatial environment. This study employs qualitative research methods such as interviews and thematic analysis, focusing on spatial environment behavior research. By analyzing the behaviors and preferences of key employment groups in China (college graduates, migrant workers, and reemployed laid-off workers) from the perspectives of physical and socio-cultural environments, this study identifies the consistencies and differences in personal viewpoints during job selection and their impact on employment decisions. The findings indicate that college graduates tend to emphasize the macro spatial environment, while migrant workers and reemployed laid-off workers are more concerned with micro spatial environment factors. Additionally, college graduates have higher requirements for the diversity of space types and the distinction between public and private spaces. Furthermore, aside from salary considerations, key employment groups generally place less importance on the socio-cultural environment compared to the physical environment. This study aims to highlight the significance of spatial environment in job selection decisions from the perspective of diverse job seekers, providing insights for policy-making and corporate recruitment strategies.

Keywords: job-seeking populations, spatial environment behavior, micro, macro, qualitative research, physical environment, socio-cultural environment

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Authors: Muhammad Saiful Islam Khan, Yun Ji Kim

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Organophosphorus pesticides (OPPs) have been widely used to replace more persistent organochlorine pesticides because OPPs are more soluble in water and decompose rapidly in aquatic systems. Extensive uses of OPPs in modern agriculture are the major cause of the contamination of surface water. Regardless of the advantages gained by the application of pesticides in modern agriculture, they are a threat to the public health environment. With the aim of reducing possible health threats, several physical and chemical treatment processes have been studied to eliminate biological and chemical poisons from food stuff. In the present study, a micro-plasma device was used to reduce pesticides from the surface of food stuff. Pesticide free food items chosen in this study were perilla leaf, tomato, broccoli and blueberry. To evaluate the removal efficiency of pesticides, different washing methods were followed such as soaking with water, washing with bubbling water, washing with plasma-treated water and washing with chlorine water. 2 mL of 2000 ppm pesticide samples, namely, diazinone and chlorpyrifos were individuality inoculated on food surface and was air dried for 2 hours before treated with plasma. Plasma treated water was used in two different manners one is plasma treated water with bubbling the other one is aerosolized plasma treated water. The removal efficiency of pesticides from food surface was studied using HPLC. Washing with plasma treated water, aerosolized plasma treated water and chlorine water shows minimum 72% to maximum 87 % reduction for 4 min treatment irrespective to the types of food items and the types of pesticides sample, in case of soaking and bubbling the reduction is 8% to 48%. Washing with plasma treated water, aerosolized plasma treated water and chlorine water shows somewhat similar reduction ability which is significantly higher comparing to the soaking and bubbling washing system. The temperature effect of the washing systems was also evaluated; three different temperatures were set for the experiment, such as 22°C, 10°C and 4°C. Decreasing temperature from 22°C to 10°C shows a higher reduction in the case of washing with plasma and aerosolized plasma treated water, whereas an opposite trend was observed for the washing with chlorine water. Further temperature reduction from 10°C to 4°C does not show any significant reduction of pesticides, except for the washing with chlorine water. Chlorine water treatment shows lesser pesticide reduction with the decrease in temperature. The color changes of the treated sample were measured immediately and after one week to evaluate if there is any effect of washing with plasma treated water and with chlorine water. No significant color changes were observed for either of the washing systems, except for broccoli washing with chlorine water.

Keywords: chlorpyrifos, diazinone, pesticides, micro plasma

Procedia PDF Downloads 178

Authors: S. Reshma Reghu, Shiburaj Sugathan, T. G. Nandu, K. B. Ramesh Kumar, Mathew Dan

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Bacterial diseases are considered to be one of the most prevalent health hazards in the developing world and many bacteria are becoming resistant to existing antibiotics making the treatment ineffective. Thus, it is necessary to find novel targets and develop new antibacterial drugs with a novel mechanism of action. The process of bacterial cell division is a novel and attractive target for new antibacterial drug discovery. FtsZ, a homolog of eukaryotic tubulin, is the major protein of the bacterial cell division machinery and is considered as an important antibacterial drug target. Zingiberaceae, the Ginger family consists of aromatic herbs with creeping rhizomes. Many of these plants have antimicrobial properties.This study aimed to determine the anti-bacterial activity of selected Zingiberaceous plants by targeting bacterial cell division protein, FtsZ. Essential oils and methanol extracts of Amomum ghaticum, Alpinia galanga, Kaempferia galanga, K. rotunda, and Zingiber officinale were tested to find its antibacterial efficiency using disc diffusion method against authentic bacterial strains obtained from MTCC (India). Essential oil isolated from A.galanga and Z.officinale were further assayed for FtsZ inhibition assay following non-radioactive malachite green-phosphomolybdate assay using E. coli FtsZ protein obtained from Cytoskelton Inc., USA. Z.officinale essential oil possess FtsZ inhibitory property. A molecular docking study was conducted with the known bioactive compounds of Z. officinale as ligands with the E. coli FtsZ protein homology model. Some of the major constituents of this plant like catechin, epicatechin, and gingerol possess agreeable docking scores. The results of this study revealed that several chemical constituents in Ginger plants can be utilised as potential source of antibacterial activity and it can warrant further investigation through drug discovery studies.

Keywords: antibacterial, FtsZ, zingiberaceae, docking

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Authors: Bharat Kumar, Deepak Kumar, Vijay Chaudhry

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Zirconium alloys are widely used for the core components of a pressurized heavy water reactor (PHWR) or Canada deuterium (CANDU) reactor due to their low neutron absorption cross-section and excellent mechanical properties. The components made of Zirconium alloys are subjected to flow-induced vibrations, resulting in fretting wear at the interface of; pressure tubes and bearing pads, pressure tubes and calandria tubes, and calandria tubes and Liquid injection shutdown system (LISS) nozzles. There is a need to explore the tribological response under such conditions. Present work simulates the contact between calandria tube and LISS nozzle of PHWR/CANDU reactor as cylinder-on-cylinder contact configuration. Reciprocating tribo-tests were conducted on Zircaloy-4 (Zr-4) under the self-mated condition at varying amplitude, frequency, and sliding time. To understand the active wear mechanism, worn surfaces were analyzed using Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). The change in amplitude severely affects the wear than other factors. The wear mechanism transits from adhesion to abrasion with increasing test amplitude. The dominant wear mechanisms are micro-cutting and micro-plowing followed by delamination in some areas. However, the coefficient of friction has indifferent behaviors.

Keywords: zircaloy-4, tribology, calandria tube, LISS nozzle, PHWR

Procedia PDF Downloads 193

Authors: Zipei Yin

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Community space is the main place for children's daily outdoor activities. The mountain community space has the typical characteristics of a closed natural environment, a scattered population layout with height differences, and a relatively independent group structure. This has resulted in special limitations on children's outdoor activities in terms of safety, accessibility, and appropriateness, which urgently makes it necessary to explore how to construct children's activity spaces in mountainous societies under the special limitations. This study investigated the activity spaces for children aged 3-11 years old in typical old communities in Chongqing and evaluated them based on the dimensions of spatial characteristics, environmental safety, and connectivity to summarise three typical patterns of children's outdoor activity spaces in old communities in mountainous cities. Then, under the framework of the appeal of the child-friendly urban environment, taking advantage of the characteristics of the old community in mountain cities compared with the plain urban community, such as complex social form, diversified functional positioning, and good foundation of autonomy, this paper explores the micro-renewal path and strategy of the compound utilization of community public space from the two levels of design and governance, so as to further promote the research and practice of the healthy development of mountain urban community environment.

Keywords: child-friendly, healthy community, community public space, mountainous urban community, community renewal

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Authors: Rasha Saad, Jean Frederic Weber, Fatimah Bebe, Sadia Sultan

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The goal of study was to screen the effects of candesartan and four endophytic fungi for their potential in microbial biotransformation. In this experiment, four types of unidentified fungi with the codes of TH2L1, TH2R10, TH1P35 and TH1S46 were used in screening process by MECFUS (Microtiter plate, Elicitors, Combination, Freeze-drying, UHPLC, Statistical analysis) protocol. The experiment was carried out by using 96-well microtiter plate (MTP) with different media and elicitors. Various media with two concentrations of Potato Dextrose Broth (PDB) and elicitors used were to induce the production of secondary metabolites from the fungi as well as the biotransformation of the drug compound. After incubation, cultures were extracted by freeze drying method and finally analyzed by ultra-High performance Liquid Chromatography (uHPLC). The extracts analyzed by uHPLC followed by LC/Ms, demonstrated the presence of biotransformation products from the drug compound and elicitation of the secondary metabolism from the fungi by the occurrence of the additional peaks. From the four fungi, TH1S46 showed highly potential produced secondary metabolites as well as the biotransformation of candesartan. For other fungi, they responded when candesartan was introduced. Moreover, the additional peaks produced in uHPLC need to be further investigation by using LC-MS or NMR.

Keywords: biotransformation, candesartan, endophytes, secondary metabolites

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Authors: Lynette Lincoln, Sunil S. More

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With the rising demand of sugar used today, it is proposed that world sugar is expected to escalate up to 203 million tonnes by 2021. Hydrolysis of sucrose (table sugar) into glucose and fructose equimolar mixture is catalyzed by β-D-fructofuranoside fructohydrolase (EC 3.2.1.26), commonly called as invertase. For fluid filled center in chocolates, preparation of artificial honey, as a sweetener and especially to ensure that food stuffs remain fresh, moist and soft for longer spans invertase is applied widely and is extensively being used. From an industrial perspective, properties such as increased solubility, osmotic pressure and prevention of crystallization of sugar in food products are highly desired. Screening for invertase does not involve plate assay/qualitative test to determine the enzyme production. In this study, we use a three-step screening strategy for identification of a novel bacterial isolate from soil which is positive for invertase production. The primary step was serial dilution of soil collected from sugarcane fields (black soil, Maddur region of Mandya district, Karnataka, India) was grown on a Czapek-Dox medium (pH 5.0) containing sucrose as the sole C-source. Only colonies with the capability to utilize/breakdown sucrose exhibited growth. Bacterial isolates released invertase in order to take up sucrose, splitting the disaccharide into simple sugars. Secondly, invertase activity was determined from cell free extract by measuring the glucose released in the medium at 540 nm. Morphological observation of the most potent bacteria was examined by several identification tests using Bergey’s manual, which enabled us to know the genus of the isolate to be Bacillus. Furthermore, this potent bacterial colony was subjected to 16S rDNA PCR amplification and a single discrete PCR amplicon band of 1500 bp was observed. The 16S rDNA sequence was used to carry out BLAST alignment search tool of NCBI Genbank database to obtain maximum identity score of sequence. Molecular sequencing and identification was performed by Xcelris Labs Ltd. (Ahmedabad, India). The colony was identified as Bacillus sp. BAB-3434, indicating to be the first novel strain for extracellular invertase production. Molasses, a by-product of the sugarcane industry is a dark viscous liquid obtained upon crystallization of sugar. An enhanced invertase production and optimization studies were carried out by one-factor-at-a-time approach. Crucial parameters such as time course (24 h), pH (6.0), temperature (45 °C), inoculum size (2% v/v), N-source (yeast extract, 0.2% w/v) and C-source (molasses, 4% v/v) were found to be optimum demonstrating an increased yield. The findings of this study reveal a simple screening method of an extracellular invertase from a rapidly growing Bacillus sp., and selection of best factors that elevate enzyme activity especially utilization of molasses which served as an ideal substrate and also as C-source, results in a cost-effective production under submerged conditions. The invert mixture could be a replacement for table sugar which is an economic advantage and reduce the tedious work of sugar growers. On-going studies involve purification of extracellular invertase and determination of transfructosylating activity as at high concentration of sucrose, invertase produces fructooligosaccharides (FOS) which possesses probiotic properties.

Keywords: Bacillus sp., invertase, molasses, screening, submerged fermentation

Procedia PDF Downloads 224

Authors: Mohamed Loutou, Mohamed Hajjaji, Mohamed Ait Babram, Mohammed Mansori, Rachid Hakkou, Claude Favotto

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More than 26,4 million tons of phosphates are produced by the phosphates industries in Morocco (2010), generating huge amounts of sludge by flocculation during the ore beneficiation. They way are stored at the end of the process in open air ponds. Its accumulation and storage may have an impact on several scales such as ground water and human being. For this purpose, an efficient way to use it the field of the ceramic is proposed. The as received sludge and a clay-rich sediment have been studied in terms of chemical, mineralogical and micro-structural side using various analytical methods. Several formulations have been performed by mixing the sludge with the binder shaped in the form of granules. After being dried at 105 °C, the samples were heated in the range of 900-1200 °C. As well as the ceramic properties (firing shrinkage, water absorption, total porosity and compressive strength) the micro structure has been investigated using X-ray diffraction, scanning electron microscopy and Fourier transform infrared spectroscopy. The relations between properties and the operating factors were formulated using the design of experiments (DOE). Gehlenite was the only phase neo-formed in the sintering samples. SEM micrographs revealed the presence of nano metric stains. Based on RSM results, all factors had positive effects on Firing shrinkage, compressive strength and total porosity. However, they manifested opposite effects on density and water absorption.

Keywords: phosphate sludge, clay, ceramic properties, granule

Procedia PDF Downloads 498

Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohammed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohammed Hussein Ayoub

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Many herbal medicinal products are considered as potential hypocholesterolemic agents with encouraging safety profiles, however, only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolaemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata peel, Zingiber officinale rhizome, and Sesamum indicum seeds. Forty-five rats were used throughout the experiment, which were divided into nine groups, five rats in each as follows; normal control group (normal rats fed with standard normal diet), rats fed hypercholesterolemic diet consisting of 1% cholesterol and 10% saturated animal fat, which were further divided into eight groups; hypercholesterolemic control group (rats only fed hypercholesterolemic diet), groups 3,4,5,6,7, and 8 were given Citrus reticulata, Zingiber officinale, and Sesamum indicum ethanolic extracts at doses of (250mg/kg and 500mg/kg, respectively) orally; and group 9 rats were given atorvastatin (0.18mg/kg) orally as a reference antihypercholesterolaemic drug. Blood samples were obtained four weeks following treatment from the retro-orbital venous plexus after fasting overnight from all groups and the lipid profile (serum total cholesterol (TC), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and triglycerides levels) was measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plant extracts was determined using DPPH free-radical assay. Results of in vivo and in vitro antihypercholesterolaemic and antioxidant assay, respectively, revealed that the three extracts possess comparable antioxidant and anti-hypercholesterolaemic activities.

Keywords: anti hypercholesterolemic effects, antioxidant activity, HDL, LDL, TC, TGs, citrus reticulata, sesamum indicum, zingiber officinale

Procedia PDF Downloads 450

Authors: Shama Ranjan Barua, Tofazzal M. Rakib, Mohammad Alamgir Hossain, Tania Ferdushy, Sharmin Chowdhury

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Rotavirus is one of the main etiologies of neonatal diarrhea in bovine calves that causes significant economic loss in Bangladesh. The present study was carried out to investigate the pathology of neonatal enteritis in calves due to bovine rotavirus infection in south-eastern part of Bangladesh. Rotavirus was identified by using ELISA, RT-PCR (Reverse Transcription Polymerase Chain Reaction), real-time RT-PCR. We examined 12 dead calves with history of diarrhea during necropsy. Among 12 dead calves, in gross examination, 6 were found with pathological changes in intestine, 5 calves had congestion of small intestine and rest one had no distinct pathological changes. Intestinal contents and/or faecal samples of all dead calves were collected and examined to confirm the presence of bovine rotavirus A using Enzyme linked immunosorbant assay (ELISA), RT-PCR and real-time RT-PCR. Out 12 samples, 5 (42%) samples revealed presence of bovine rotavirus A in three diagnostic tests. The histopathological changes were found almost exclusively limited in the small intestine. The lesions of rotaviral enteritis ranged from slight to moderate shortening (atrophy) of villi in the jejunum and ileum with necrotic crypts. The villi were blunt and covered by immature epithelial cells. Infected cells, stained with Haematoxylin and Eosin staining method, showed characteristic syncytia and eosinophilc intracytoplasmic inclusion body. The presence of intracytoplasmic inclusion bodies in enterocytes is the indication of viral etiology. The presence of rotavirus in the affected tissues and/or lesions was confirmed by three different immunological and molecular tests. The findings of histopathological changes will be helpful in future diagnosis of rotaviral infection in dead calves.

Keywords: calves, diarrhea, pathology, rotavirus

Procedia PDF Downloads 244

Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

Procedia PDF Downloads 161

Authors: Jun Hyung Lee, Robin B. Guevarra, Jin Ho Cho, Bo-Ra Kim, Jiwon Shin, Doo Wan Kim, Young Hwa Kim, Minho Song, Hyeun Bum Kim

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Salmonella is one of the most important swine pathogens, causing acute or chronic digestive diseases, such as enteritis. The acute form of enteritis is common in young pigs of 2-4 months of age. Salmonellosis in swine causes a huge economic burden to swine industry by reducing production. Therefore, it is necessary that swine industries should strive to decrease Salmonellosis in pigs in order to reduce economic losses. Thus, we tested three types of natural plant extracts(PEs) to evaluate antibacterial effects against Salmonella enterica Typhimurium isolated from the piglet with Salmonellosis. Three PEs including turmeric oleoresin (containing curcumin 79 to 85%), capsicum oleoresin (containing capsaicin 40%-40.1%), and garlic essential oil (100% natural garlic) were tested using the direct contact agar diffusion test, minimum inhibitory concentration test, growth curve assay, and heat stability test. The tests were conducted with PEs at each concentration of 2.5%, 5%, and 10%. For the heat stability test, PEs with 10% concentration were incubated at each 4, 20, 40, 60, 80, and 100 °C for 1 hour; then the direct contact agar diffusion test was used. For the positive and negative controls, 0.5N HCl and 1XPBS were used. All the experiments were duplicated. In the direct contact agar diffusion test, garlic essential oil with 2.5%, 5%, and 10% concentration showed inhibit zones of 1.5cm, 2.7cm, and 2.8cm diameters compared to that of 3.5cm diameter for 0.5N HCl. The minimum inhibited concentration of garlic essential oil was 2.5%. Growth curve assay showed that the garlic essential oil was able to inhibit Salmonella growth significantly after 4hours. The garlic essential oil retained the ability to inhibit Salmonella growth after heat treatment at each temperature. However, turmeric and capsicum oleoresins were not able to significantly inhibit Salmonella growth by all the tests. Even though further in-vivo tests will be needed to verify effects of garlic essential oil for the Salmonellosis prevention for piglets, our results showed that the garlic essential oil could be used as a potential natural agent to prevent Salmonellosis in swine.

Keywords: garlic essential oil, pig, salmonellosis, Salmonella enterica

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