Search results for: substrate roughness
251 Perinatal Ethanol Exposure Modifies CART System in Rat Brain Anticipated for Development of Anxiety, Depression and Memory Deficits
Authors: M. P. Dandekar, A. P. Bharne, P. T. Borkar, D. M. Kokare, N. K. Subhedar
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Ethanol ingestion by the mother ensue adverse consequences for her offspring. Herein, we examine the behavioral phenotype and neural substrate of the offspring of the mother on ethanol. Female rats were fed with ethanol-containing liquid diet from 8 days prior of conception and continued till 25 days post-parturition to coincide with weaning. Behavioral changes associated with anxiety, depression and learning and memory were assessed in the offspring, after they attained adulthood (day 85), using elevated plus maze (EPM), forced swim (FST) and novel object recognition tests (NORT), respectively. The offspring of the alcoholic mother, compared to those of the pair-fed mother, spent significantly more time in closed arms of EPM and showed more immobility time in FST. Offspring at the age of 25 and 85 days failed to discriminate between novel versus familiar object in NORT, thus reflecting anxiogenic, depressive and amnesic phenotypes. Neuropeptide cocaine- and amphetamine-regulated transcript peptide (CART) is known to be involved in central effects of ethanol and hence selected for the current study. Twenty-five days old pups of the alcoholic mother showed significant augmentation in CART-immunoreactivity in the cells of Edinger-Westphal (EW) nucleus and lateral hypothalamus. However, a significant decrease in CART-immunoreactivity was seen in nucleus accumbens shell (AcbSh), lateral part of bed nucleus of the stria terminalis (BNSTl), locus coeruleus (LC), hippocampus (CA1, CA2 and CA3), and arcuate nucleus (ARC) of the pups and/or adults offspring. While no change in the CART-immunoreactive fibers of AcbSh and BNSTl, CA2 and CA3 was noticed in the 25 days old pups, the CART-immunoreactive cells in EW and paraventricular nucleus (PVN), and fibers in the central nucleus of amygdala of 85 days old offspring remained unaffected. We suggest that the endogenous CART system in these discrete areas, among other factors, may be a causal to the abnormalities in the next generation of an alcoholic mother.Keywords: anxiety, depression, CART, ethanol, immunocytochemistry
Procedia PDF Downloads 395250 An Approach to Study the Biodegradation of Low Density Polyethylene Using Microbial Strains of Bacillus subtilus, Aspergillus niger, Pseudomonas fluroscence in Different Media Form and Salt Condition
Authors: Monu Ojha, Rahul Rana, Satywati Sharma, Kavya Dashora
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The global production rate of plastics has increased enormously and global demand for polyethylene resins –High-density polyethylene (HDPE), Linear low-density polyethylene (LLDPE) and Low-density polyethylene (LDPE) is expected to rise drastically, with very high value. These get accumulated in the environment, posing a potential ecological threat as they are degrading at a very slow rate and remain in the environment indefinitely. The aim of the present study was to investigate the potential of commonly found soil microbes like Bacillus subtilus, Aspergillus niger, Pseudomonas fluroscence for their ability to biodegrade LDPE in the lab on solid and liquid media conditions as well as in presence of 1% salt in the soil. This study was conducted at Indian Institute of Technology, Delhi, India from July to September where average temperature and RH (Relative Humidity) were 33 degrees Celcius and 80% respectively. It revealed that the weight loss of LDPE strip obtained from market of approximately 4x6 cm dimensions is more in liquid broth media than in solid agar media. The percentage weight loss by P. fluroscence, A. niger and B. subtilus observed after 80 days of incubation was 15.52, 9.24 and 8.99% respectively in broth media and 6.93, 2.18 and 4.76 % in agar media. The LDPE strips from same source and on the same were subjected to soil in presence of above microbes with 1% salt (NaCl: obtained from commercial table salt) with temperature and RH 33 degree Celcius and 80%. It was found that the rate of degradation increased in the soil than under lab conditions. The rate of weight loss of LDPE strips under same conditions given in lab was found to be 32.98, 15.01 and17.09 % by P. fluroscence, A. niger and B. subtilus respectively. The breaking strength was found to be 9.65N, 29N and 23.85 N for P. fluroscence, A. niger and B. subtilus respectively. SEM analysis conducted on Zeiss EVO 50 confirmed that surface of LDPE becomes physically weak after biological treatment. There was the increase in the surface roughness indicating Surface erosion of LDPE film. FTIR (Fourier-transform infrared spectroscopy) analysis of the degraded LDPE films showed stretching of aldehyde group at 3334.92 and 3228.84 cm-1,, C–C=C symmetric of aromatic ring at 1639.49 cm-1.There was also C=O stretching of aldehyde group at 1735.93 cm-1. N=O peak bend was also observed which corresponds to 1365.60 cm-1, C–O stretching of ether group at 1217.08 and 1078.21 cm-1.Keywords: microbial degradation, LDPE, Aspergillus niger, Bacillus subtilus, Peudomonas fluroscence, common salt
Procedia PDF Downloads 165249 Short-Term Impact of a Return to Conventional Tillage on Soil Microbial Attributes
Authors: Promil Mehra, Nanthi Bolan, Jack Desbiolles, Risha Gupta
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Agricultural practices affect the soil physical and chemical properties, which in turn influence the soil microorganisms as a function of the soil biological environment. On the return to conventional tillage (CT) from continuing no-till (NT) cropping system, a very little information is available from the impact caused by the intermittent tillage on the soil biochemical properties from a short-term (2-year) study period. Therefore, the contribution made by different microorganisms (fungal, bacteria) was also investigated in order to find out the effective changes in the soil microbial activity under a South Australian dryland faring system. This study was conducted to understand the impact of microbial dynamics on the soil organic carbon (SOC) under NT and CT systems when treated with different levels of mulching (0, 2.5 and 5 t/ha). Our results demonstrated that from the incubation experiment the cumulative CO2 emitted from CT system was 34.5% higher than NT system. Relatively, the respiration from surface layer (0-10 cm) was significantly (P<0.05) higher by 8.5% and 15.8 from CT; 8% and 18.9% from NT system w.r.t 10-20 and 20-30 cm respectively. Further, the dehydrogenase enzyme activity (DHA) and microbial biomass carbon (MBC) were both significantly lower (P<0.05) under CT, i.e., 7.4%, 7.2%, 6.0% (DHA) and 19.7%, 15.7%, 4% (MBC) across the different mulching levels (0, 2.5, 5 t/ha) respectively. In general, it was found that from both the tillage system the enzyme activity and MBC decreased with the increase in depth (0-10, 10-20 and 20-30 cm) and with the increase in mulching rate (0, 2.5 and 5 t/ha). From the perspective of microbial stress, there was 28.6% higher stress under CT system compared to NT system. Whereas, the microbial activity of different microorganisms like fungal and bacterial activities were determined by substrate-induced inhibition respiration using antibiotics like cycloheximide (16 mg/gm of soil) and streptomycin sulphate (14 mg/gm of soil), by trapping the CO2 using an alkali (0.5 M NaOH) solution. The microbial activities were confirmed through platting technique, where it was that found bacterial activities were 46.2% and 38.9% higher than fungal activity under CT and NT system. In conclusion, it was expected that changes in the relative abundance and activity of different microorganisms (bacteria and fungi) under different tillage systems could significantly affect the C cycling and storage due to its unique structures and differential interactions with the soil physical properties.Keywords: tillage, soil respiration, MBC, fungal-bacterial activity
Procedia PDF Downloads 261248 Ambient Electrospray Deposition: An Efficient Technique to Immobilize Laccase on Cheap Electrodes With Unprecedented Reuse and Storage Performances
Authors: Mattea Carmen Castrovilli, Antonella Cartoni
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Electrospray ionisation (ESI), a well-established technique widely used to produce ion beams of biomolecules in mass spectrometry (ESI-MS), can be used for ambient soft landing of enzymes on a specific substrate. In this work, we show how the ambient electrospray deposition (ESD) technique can be successfully exploited for manufacturing a promising, green-friendly electrochemical amperometric laccase-based biosensor with unprecedented reuse and storage performance. These biosensors have been manufactured by spraying a laccase solution of 2μg/μL at 20% of methanol on a commercial carbon screen printed electrode (C-SPE) using a custom ESD set-up. The laccase-based ESD biosensor has been tested against catechol compounds in the linear range 2-100 μM, with a limit of detection of 1.7 μM, without interference from cadmium, chrome, arsenic, and zinc and without any memory effects, but showing a matrix effect in lake and well water. The ESD biosensor shows enhanced performances compared to the ones fabricated with other immobilization methods, like drop-casting. Indeed, it retains 100% activity up to two months of storage at ambient conditions without any special care and working stability up to 63 measurements on the same electrode just prepared and 20 on a one-year-old electrode subjected to redeposition together with a 100% resistance to use of the same electrode in subsequent days. The ESD method is a one-step, environmentally friendly method that allows the deposition of the bio-recognition layer without using any additional chemicals. The promising results in terms of storage and working stability also obtained with the more fragile lactate oxidase enzyme suggest these improvements should be attributed to the ESD technique rather than to the bioreceptor, highlighting how the ESD could be useful in reducing pollution from disposable devices. Acknowledgment: The understanding at the molecular level of this promising biosensor by using different spectroscopies, microscopies and analytical techniques is the subject of our PRIN 2022 project ESILARANTE.Keywords: reuse, storage performance, immobilization, electrospray deposition, biosensor, laccase, catechol detection, green chemistry
Procedia PDF Downloads 62247 An Investigation on the Pulse Electrodeposition of Ni-TiO2/TiO2 Multilayer Structures
Authors: S. Mohajeri
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Electrocodeposition of Ni-TiO2 nanocomposite single layers and Ni-TiO2/TiO2 multilayers from Watts bath containing TiO2 sol was carried out on copper substrate. Pulse plating and pulse reverse plating techniques were applied to facilitate higher incorporations of TiO2 nanoparticles in Ni-TiO2 nanocomposite single layers, and the results revealed that by prolongation of the current-off durations and the anodic cycles, deposits containing 11.58 wt.% and 13.16 wt.% TiO2 were produced, respectively. Multilayer coatings which consisted of Ni-TiO2 and TiO2-rich layers were deposited by pulse potential deposition through limiting the nickel deposition by diffusion control mechanism. The TiO2-rich layers thickness and accordingly, the content of TiO2 reinforcement reached 104 nm and 18.47 wt.%, respectively in the optimum condition. The phase structure and surface morphology of the nanocomposite coatings were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The cross sectional morphology and line scans of the layers were studied by field emission scanning electron microscopy (FESEM). It was confirmed that the preferred orientations and the crystallite sizes of nickel matrix were influenced by the deposition technique parameters, and higher contents of codeposited TiO2 nanoparticles refined the microstructure. The corrosion behavior of the coatings in 1M NaCl and 0.5M H2SO4 electrolytes were compared by means of potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) techniques. Increase of corrosion resistance and the passivation tendency were favored by TiO2 incorporation, while the degree of passivation declined as embedded particles disturbed the continuity of passive layer. The role of TiO2 incorporation on the improvement of mechanical properties including hardness, elasticity, scratch resistance and friction coefficient was investigated by the means of atomic force microscopy (AFM). Hydrophilicity and wettability of the composite coatings were investigated under UV illumination, and the water contact angle of the multilayer was reduced to 7.23° after 1 hour of UV irradiation.Keywords: electrodeposition, hydrophilicity, multilayer, pulse-plating
Procedia PDF Downloads 249246 River Habitat Modeling for the Entire Macroinvertebrate Community
Authors: Pinna Beatrice., Laini Alex, Negro Giovanni, Burgazzi Gemma, Viaroli Pierluigi, Vezza Paolo
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Habitat models rarely consider macroinvertebrates as ecological targets in rivers. Available approaches mainly focus on single macroinvertebrate species, not addressing the ecological needs and functionality of the entire community. This research aimed to provide an approach to model the habitat of the macroinvertebrate community. The approach is based on the recently developed Flow-T index, together with a Random Forest (RF) regression, which is employed to apply the Flow-T index at the meso-habitat scale. Using different datasets gathered from both field data collection and 2D hydrodynamic simulations, the model has been calibrated in the Trebbia river (2019 campaign), and then validated in the Trebbia, Taro, and Enza rivers (2020 campaign). The three rivers are characterized by a braiding morphology, gravel riverbeds, and summer low flows. The RF model selected 12 mesohabitat descriptors as important for the macroinvertebrate community. These descriptors belong to different frequency classes of water depth, flow velocity, substrate grain size, and connectivity to the main river channel. The cross-validation R² coefficient (R²𝒸ᵥ) of the training dataset is 0.71 for the Trebbia River (2019), whereas the R² coefficient for the validation datasets (Trebbia, Taro, and Enza Rivers 2020) is 0.63. The agreement between the simulated results and the experimental data shows sufficient accuracy and reliability. The outcomes of the study reveal that the model can identify the ecological response of the macroinvertebrate community to possible flow regime alterations and to possible river morphological modifications. Lastly, the proposed approach allows extending the MesoHABSIM methodology, widely used for the fish habitat assessment, to a different ecological target community. Further applications of the approach can be related to flow design in both perennial and non-perennial rivers, including river reaches in which fish fauna is absent.Keywords: ecological flows, macroinvertebrate community, mesohabitat, river habitat modeling
Procedia PDF Downloads 94245 Tribological Behaviour of the Degradation Process of Additive Manufactured Stainless Steel 316L
Authors: Yunhan Zhang, Xiaopeng Li, Zhongxiao Peng
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Additive manufacturing (AM) possesses several key characteristics, including high design freedom, energy-efficient manufacturing process, reduced material waste, high resolution of finished products, and excellent performance of finished products. These advantages have garnered widespread attention and fueled rapid development in recent decades. AM has significantly broadened the spectrum of available materials in the manufacturing industry and is gradually replacing some traditionally manufactured parts. Similar to components produced via traditional methods, products manufactured through AM are susceptible to degradation caused by wear during their service life. Given the prevalence of 316L stainless steel (SS) parts and the limited research on the tribological behavior of 316L SS samples or products fabricated using AM technology, this study aims to investigate the degradation process and wear mechanisms of 316L SS disks fabricated using AM technology. The wear mechanisms and tribological performance of these AM-manufactured samples are compared with commercial 316L SS samples made using conventional methods. Additionally, methods to enhance the tribological performance of additive-manufactured SS samples are explored. Four disk samples with a diameter of 75 mm and a thickness of 10 mm are prepared. Two of them (Group A) are prepared from a purchased SS bar using a milling method. The other two disks (Group B), with the same dimensions, are made of Gas Atomized 316L Stainless Steel (size range: 15-45 µm) purchased from Carpenter Additive and produced using Laser Powder Bed Fusion (LPBF). Pin-on-disk tests are conducted on these disks, which have similar surface roughness and hardness levels. Multiple tests are carried out under various operating conditions, including varying loads and/or speeds, and the friction coefficients are measured during these tests. In addition, the evolution of the surface degradation processes is monitored by creating moulds of the wear tracks and quantitatively analyzing the surface morphologies of the mould images. This analysis involves quantifying the depth and width of the wear tracks and analyzing the wear debris generated during the wear processes. The wear mechanisms and wear performance of these two groups of SS samples are compared. The effects of load and speed on the friction coefficient and wear rate are investigated. The ultimate goal is to gain a better understanding of the surface degradation of additive-manufactured SS samples. This knowledge is crucial for enhancing their anti-wear performance and extending their service life.Keywords: degradation process, additive manufacturing, stainless steel, surface features
Procedia PDF Downloads 79244 Investigation of the Growth Kinetics of Phases in Ni–Sn System
Authors: Varun A Baheti, Sanjay Kashyap, Kamanio Chattopadhyay, Praveen Kumar, Aloke Paul
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Ni–Sn system finds applications in the microelectronics industry, especially with respect to flip–chip or direct chip, attach technology. Here the region of interest is under bump metallization (UBM), and solder bump (Sn) interface due to the formation of brittle intermetallic phases there. Understanding the growth of these phases at UBM/Sn interface is important, as in many cases it controls the electro–mechanical properties of the product. Cu and Ni are the commonly used UBM materials. Cu is used for good bonding because of fast reaction with solder and Ni often acts as a diffusion barrier layer due to its inherently slower reaction kinetics with Sn–based solders. Investigation on the growth kinetics of phases in Ni–Sn system is reported in this study. Just for simplicity, Sn being major solder constituent is chosen. Ni–Sn electroplated diffusion couples are prepared by electroplating pure Sn on Ni substrate. Bulk diffusion couples prepared by the conventional method are also studied along with Ni–Sn electroplated diffusion couples. Diffusion couples are annealed for 25–1000 h at 50–215°C to study the phase evolutions and growth kinetics of various phases. The interdiffusion zone was analysed using field emission gun equipped scanning electron microscope (FE–SEM) for imaging. Indexing of selected area diffraction (SAD) patterns obtained from transmission electron microscope (TEM) and composition measurements done in electron probe micro−analyser (FE–EPMA) confirms the presence of various product phases grown across the interdiffusion zone. Time-dependent experiments indicate diffusion controlled growth of the product phase. The estimated activation energy in the temperature range 125–215°C for parabolic growth constants (and hence integrated interdiffusion coefficients) of the Ni₃Sn₄ phase shed light on the growth mechanism of the phase; whether its grain boundary controlled or lattice controlled diffusion. The location of the Kirkendall marker plane indicates that the Ni₃Sn₄ phase grows mainly by diffusion of Sn in the binary Ni–Sn system.Keywords: diffusion, equilibrium phase, metastable phase, the Ni-Sn system
Procedia PDF Downloads 307243 Study of the Hysteretic I-V Characteristics in a Polystyrene/ZnO-Nanorods Stack Layer
Authors: You-Lin Wu, Yi-Hsing Sung, Shih-Hung Lin, Jing-Jenn Lin
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Performance improvement in optoelectronic devices such as solar cells and photodetectors has been reported when a polymer/ZnO nanorods stack is used. Resistance switching of polymer/ZnO nanocrystals (or nanorods) hybrid has also gained a lot of research interests recently. It has been reported that high- and low-resistance states of a metal/insulator/metal (MIM) structure diode with a polystyrene (PS) and ZnO hybrid as the insulator layer can be switched by applied bias after a high-voltage forming process, while the same device structure merely with a PS layer does not show any forming behavior. In this work, we investigated the current-voltage (I-V) characteristics of an MIM device with a PS/ZnO nanorods stack deposited on fluorine-doped tin oxide (FTO) glass substrate. The ZnO nanorods were grown by a hydrothermal method using a mixture of zinc nitrate, hexamethylenetetramine, and DI water. Following that, a PS layer was deposited by spin coating. Finally, the device with a structure of Ti/ PS/ZnO nanorods/FTO was completed by e-gun evaporated Ti layer on top of the PS layer. Semiconductor parameters analyzer Agilent 4156C was then used to measure the I-V characteristics of the device by applying linear ramp sweep voltage with sweep sequence of 0V → 4V → 0V → 3V → 0V → 2V → 0V → 1V → 0V in both positive and negative directions. It is interesting to find that the I-V characteristics are bias dependent and hysteretic, indicating that the device Ti/PS/ZnO nanorods/FTO structure has ferroelectricity. Our results also show that the maximum hysteresis loop height of the I-V characteristics as well as the voltage at which the maximum hysteresis loop height of each scan occurs increase with increasing maximum sweep voltage. It should be noticed that, although ferroelectricity has been found in ZnO at its melting temperature (1975℃) and in Li- or Co-doped ZnO, neither PS nor ZnO has ferroelectricity at room temperature. Using the same structure but with a PS or ZnO layer only as the insulator does not give and hysteretic I-V characteristics. It is believed that a charge polarization layer is induced near the PS/ZnO nanorods stack interface and thus causes the ferroelectricity in the device with Ti/PS/ZnO nanorods/FTO structure. Our results show that the PS/ZnO stack can find a potential application in a resistive switching memory device with MIM structure.Keywords: ferroelectricity, hysteresis, polystyrene, resistance switching, ZnO nanorods
Procedia PDF Downloads 311242 Developing a Thermo-Sensitive Conductive Stretchable Film to Allow Cell Sheet Harvest after Mechanical and Electrical Treatments
Authors: Wei-Wen Hu, Yong-Zhi Zhong
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Depositing conductive polypyrrole (PPy) onto elastic polydimethylsiloxane (PDMS) substrate can obtain a highly stretchable conductive film, which can be used to construct a bioreactor to cyclically stretch and electrically stimulate surface cells. However, how to completely harvest these stimulated muscle tissue to repair damaged muscle is a challenge. To address this concern, N-isopropylacrylamide (NIPAAm), a monomer of temperature-sensitive polymer, was added during the polymerization of pyrrole on PDMS so that the resulting P(Py-co-NIPAAm)/PDMS should own both conductivity and thermo-sensitivity. Therefore, cells after stimulation can be completely harvested as cell sheets by reducing temperature. Mouse skeletal myoblast, C2C12 cells, were applied to examine our hypothesis. In electrical stimulation, C2C12 cells on P(Py-co-NIPAAm)/PDMS demonstrated the best myo-differentiation under the electric field of 1 V/cm. Regarding cyclic stretching, the strain equal to or higher than 9% can highly align C2C12 perpendicular to the stretching direction. The Western blotting experiments demonstrated that the cell sheets harvested by cooling reserved more extracellular matrix (ECM) than cells collected by the traditional trypsin digestion method. Immunostaining of myosin heavy chain protein (MHC) indicated that both mechanical and electrical stimuli effectively increased the number of myotubes and the differentiation ratio, and the myotubes can be aligned by cyclic stretching. Stimulated cell sheets can be harvested by cooling, and the alignment of myotubes was still maintained. These results suggested that the deposition of P(Py-co-NIPAAm) on PDMS can be applied to harvest intact cell sheets after cyclic stretching and electrical stimulation, which increased the feasibility of bioreactor for the application of tissue engineering and regenerative medicine.Keywords: bioreactor, cell sheet, conductive polymer, cyclic stretching, electrical stimulation, muscle tissue engineering, myogenesis, thermosensitive hydrophobicity
Procedia PDF Downloads 94241 Investigation of Electrochemical, Morphological, Rheological and Mechanical Properties of Nano-Layered Graphene/Zinc Nanoparticles Incorporated Cold Galvanizing Compound at Reduced Pigment Volume Concentration
Authors: Muhammad Abid
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The ultimate goal of this research was to produce a cold galvanizing compound (CGC) at reduced pigment volume concentration (PVC) to protect metallic structures from corrosion. The influence of the partial replacement of Zn dust by nano-layered graphene (NGr) and Zn metal nanoparticles on the electrochemical, morphological, rheological, and mechanical properties of CGC was investigated. EIS was used to explore the electrochemical nature of coatings. The EIS results revealed that the partial replacement of Zn by NGr and Zn nanoparticles enhanced the cathodic protection at reduced PVC (4:1) by improving the electrical contact between the Zn particles and the metal substrate. The Tafel scan was conducted to support the cathodic behaviour of the coatings. The sample formulated solely with Zn at PVC 4:1 was found to be dominated in physical barrier characteristics over cathodic protection. By increasing the concentration of NGr in the formulation, the corrosion potential shifted towards a more negative side. The coating with 1.5% NGr showed the highest galvanic action at reduced PVC. FE-SEM confirmed the interconnected network of conducting particles. The coating without NGr and Zn nanoparticles at PVC 4:1 showed significant gaps between the Zn dust particles. The novelty was evidenced when micrographs showed the consistent distribution of NGr and Zn nanoparticles all over the surface, which acted as a bridge between spherical Zn particles and provided cathodic protection at a reduced PVC. The layered structure of graphene also improved the physical shielding effect of the coatings, which limited the diffusion of electrolytes and corrosion products (oxides/hydroxides) into the coatings, which was reflected by the salt spray test. The rheological properties of coatings showed good liquid/fluid properties. All the coatings showed excellent adhesion but had different strength values. A real-time scratch resistance assessment showed all the coatings had good scratch resistance.Keywords: protective coatings, anti-corrosion, galvanization, graphene, nanomaterials, polymers
Procedia PDF Downloads 96240 The Strategy for Detection of Catecholamines in Body Fluids: Optical Sensor
Authors: Joanna Cabaj, Sylwia Baluta, Karol Malecha, Kamila Drzozga
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Catecholamines are the principal neurotransmitters that mediate a variety of the central nervous system functions, such as motor control, cognition, emotion, memory processing, and endocrine modulation. Dysfunctions in catecholamine neurotransmission are induced in some neurologic and neuropsychiatric diseases. Changeable neurotransmitters level in biological fluids can be a marker of several neurological disorders. Because of its significance in analytical techniques and diagnostics, sensitive and selective detection of neurotransmitters is increasingly attracting a lot of attention in different areas of bio-analysis or biomedical research. Recently, fluorescent techniques for detection of catecholamines have attracted interests due to their reasonable cost, convenient control, as well as maneuverability in biological environments. Nevertheless, with the observed need for a sensitive and selective catecholamines sensor, the development of a convenient method for this neurotransmitter is still at its basic level. The manipulation of nanostructured materials in conjunction with biological molecules has led to the development of a new class of hybrid modified biosensors in which both enhancement of charge transport and biological activity preservation may be obtained. Immobilization of biomaterials on electrode surfaces is the crucial step in fabricating electrochemical as well as optical biosensors and bioelectronic devices. Continuing systematic investigation in the manufacturing of enzyme–conducting sensitive systems, here is presented a convenient fluorescence sensing strategy for catecholamines detection based on FRET (fluorescence resonance energy transfer) phenomena observed for, i.e., complexes of Fe²⁺ and epinephrine. The biosensor was constructed using low temperature co-fired ceramics technology (LTCC). This sensing system used the catalytical oxidation of catecholamines and quench of the strong luminescence of obtained complexes due to FRET. The detection process was based on the oxidation of substrate in the presence of the enzyme–laccase/tyrosinase.Keywords: biosensor, conducting polymer, enzyme, FRET, LTCC
Procedia PDF Downloads 257239 Effect of Light Spectra, Light Intensity, and HRT on the Co-Production of Phycoerythrin and Exopolysaccharides from Poprhyridium Marinum
Authors: Rosaria Tizzani, Tomas Morosinotto, Fabrizio Bezzo, Eleonora Sforza
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Red microalga Porphyridium marinum CCAP 13807/10 has the potential to produce a broad range of commercially valuable chemicals such as PhycoErytrin (PE) and sulphated ExoPolySaccharides (EPS). Multiple abiotic factors influence the growth of Porphyridium sp., e.g. the wavelength of the light source and different cultivation strategies (one or two steps, batch, semi-, and continuous regime). The microalga of interest is cultivated in a two-step system. First, the culture grows photoautotrophically in a controlled bioreactor with pH-dependent CO2 injection, temperature monitoring, light intensity, and LED wavelength remote control in a semicontinuous mode. In the second step, the harvested biomass is subjected to mixotrophic conditions to enhance further growth. Preliminary tests have been performed to define the suitable media, salinity, pH, and organic carbon substrate to obtain the highest biomass productivity. Dynamic light and operational conditions (e.g. HRT) are evaluated to achieve high biomass production, high PE accumulation in the biomass, and high EPS release in the medium. Porphyridium marinum is able to chromatically adapt the photosynthetic apparatus to efficiently exploit the full light spectra composition. The effect of specific narrow LED wavelengths (white W, red R, green G, blue B) and a combination of LEDs (WR, WB, WG, BR, BG, RG) are identified to understand the phenomenon of chromatic adaptation under photoautotrophic conditions. The effect of light intensity, residence time, and light quality are investigated to define optimal operational strategies for full scale commercial applications. Production of biomass, phycobiliproteins, PE, EPS, EPS sulfate content, EPS composition, Chlorophyll-a, and pigment content are monitored to determine the effect of LED wavelength on the cultivation Porphyridium marinum in order to optimize the production of these multiple, highly valuable bioproducts of commercial interest.Keywords: red microalgae, LED, exopolysaccharide, phycoerythrin
Procedia PDF Downloads 108238 Validating Quantitative Stormwater Simulations in Edmonton Using MIKE URBAN
Authors: Mohamed Gaafar, Evan Davies
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Many municipalities within Canada and abroad use chloramination to disinfect drinking water so as to avert the production of the disinfection by-products (DBPs) that result from conventional chlorination processes and their consequential public health risks. However, the long-lasting monochloramine disinfectant (NH2Cl) can pose a significant risk to the environment. As, it can be introduced into stormwater sewers, from different water uses, and thus freshwater sources. Little research has been undertaken to monitor and characterize the decay of NH2Cl and to study the parameters affecting its decomposition in stormwater networks. Therefore, the current study was intended to investigate this decay starting by building a stormwater model and validating its hydraulic and hydrologic computations, and then modelling water quality in the storm sewers and examining the effects of different parameters on chloramine decay. The presented work here is only the first stage of this study. The 30th Avenue basin in Southern Edmonton was chosen as a case study, because the well-developed basin has various land-use types including commercial, industrial, residential, parks and recreational. The City of Edmonton has already built a MIKE-URBAN stormwater model for modelling floods. Nevertheless, this model was built to the trunk level which means that only the main drainage features were presented. Additionally, this model was not calibrated and known to consistently compute pipe flows higher than the observed values; not to the benefit of studying water quality. So the first goal was to complete modelling and updating all stormwater network components. Then, available GIS Data was used to calculate different catchment properties such as slope, length and imperviousness. In order to calibrate and validate this model, data of two temporary pipe flow monitoring stations, collected during last summer, was used along with records of two other permanent stations available for eight consecutive summer seasons. The effect of various hydrological parameters on model results was investigated. It was found that model results were affected by the ratio of impervious areas. The catchment length was tested, however calculated, because it is approximate representation of the catchment shape. Surface roughness coefficients were calibrated using. Consequently, computed flows at the two temporary locations had correlation coefficients of values 0.846 and 0.815, where the lower value pertained to the larger attached catchment area. Other statistical measures, such as peak error of 0.65%, volume error of 5.6%, maximum positive and negative differences of 2.17 and -1.63 respectively, were all found in acceptable ranges.Keywords: stormwater, urban drainage, simulation, validation, MIKE URBAN
Procedia PDF Downloads 296237 Characterization of AlOOH Film Containing Mg-Al Layered Double Hydroxide Prepared on Al Alloy by Steam Coating
Authors: Ai Serizawa, Kotaro Mori, Takahiro Ishizaki
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Al alloys have been used as advanced structural materials in automobile and railway industries because of excellent physical and mechanical properties such as low density, good heat conductivity, and high specific strength. Their low corrosion resistance, however, limits their use in the corrosive environment. To improve the corrosion resistance of the Al alloys, the development of a novel coating technology has been highly desirable. Chemical conversion methods using layered double hydroxide (LDH) have attracted much attention because the LDH can suppress corrosion reaction due to their trapping ability of corrosive anions such as Cl- between layers. In this presentation, we report on a novel preparation method of AlOOH film containing Mg-Al layered double hydroxide (LDH) on Al alloy by steam coating. The corrosion resistance of the composite film including LDH was especially focused. Al-Mg-Si alloy was used as the substrate. The substrates were ultrasonically cleaned in ethanol for 10 min. The cleaned substrates were set in the autoclave with a 100 mL capacity. 20 ml of ultrapure water was located at the bottom of the autoclave to produce steam. The autoclave was heated up to a temperature of 100 to 200 °C, and then held at this temperature for up to 48 h, and was subsequently cooled naturally to room temperature, resulting in the formation of anticorrosive films on Al alloys. The resultant films were characterized by XRD, FT-IR, FE-SEM and electrochemical measurements. FE-SEM image of film surface treated at 180 °C for 48 h demonstrated that needle-like nanostructure was densely formed on the surface. XRD patterns revealed that the film formed on the Al alloys by steam coating was composed of crystal AlOOH and Mg-Al LDH. The corrosion resistance of the film was evaluated using electrochemical measurements. The potentiodynamic polarization curves of the film coated and uncoated substrates of Al-Mg-Si alloy after immersion in the 5 wt% NaCl aqueous solution for 30 min revealed that the corrosion current density, jcorr, of the film coated sample decreased by more than two orders of magnitude as compared to the uncoated sample, indicating that the corrosion resistance of the substrates of Al-Mg-Si alloy were improved by the formation of the anticorrosive film via steam coating.Keywords: aluminum alloy, boehmite, corrosion resistance, steam process
Procedia PDF Downloads 289236 Nanostructured Oxide Layer by Anodization on Austenitic Stainless Steels: Structural and Corrosion Insights
Authors: Surya Prakash Gajagouni, Akram Alfantazi, Imad Barsoum
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Austenitic stainless steels are widely recognized for their exceptional corrosion resistance and mechanical properties, rendering them indispensable materials across various industries from construction to biomedical applications. However, in chloride and high temperature atmosphere it to further enhance their surface properties, anodization has emerged as a promising surface treatment technique. Anodization modifies the surface of stainless steels by creating a protective oxide layer, improving corrosion resistance and imparting additional functional characteristics. This paper explores the structural and corrosion characteristics of anodized austenitic stainless steels (AISI 304) using a two-step anodic technique. We utilized a perchloric acid-based electrolyte followed by an ammonium fluoride-based electrolyte. This sequential approach aimed to cultivate deeper and intricately self-ordered nanopore oxide arrays on a substrate made of 304 stainless steel. Electron Microscopic (SEM and TEM) images revealed nanoporous layered structures with increased length and crack development correlating with higher voltage and anodization time. Surface composition and chemical oxidation state of surface-treated SS were determined using X-ray photoelectron spectroscopy (XPS) techniques, revealing a surface layer rich in Ni and suppressed Cr, resulting in a thin film composed of Ni and Fe oxide compared to untreated SS. Electrochemical studies demonstrated enhanced corrosion resistance in a strong alkaline medium compared to untreated SS. Understanding the intricate relationship between the structural features of anodized stainless steels and their corrosion resistance is crucial for optimizing the performance of these materials in diverse applications. This study aims to contribute to the advancement of surface engineering strategies for enhancing the durability and functionality of austenitic stainless steels in aggressive environments.Keywords: austenitic stainless steel, anodization, nanoporous oxides, marine corrosion
Procedia PDF Downloads 34235 Influence of IL-1β on Hamster Blastocyst Hatching via Regulation of Hatching Associated Proteases
Authors: Madhulika Pathak, Polani Seshagiri, Vani Venkatappa
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Blastocyst hatching is an indispensable process for successful implantation. One of the major reasons for implantation failure in IVF clinic is poor quality of embryo, which are not development/hatching-competent. Therefore, attempts are required to develop or enhance the culture system with a molecule recapitulating the autocrine/paracrine factors containing the environment of in-vivo endometrial milieu. We have tried to explore the functional molecules involved in the hamster hatching phenomenon. Blastocyst hatching is governed by several molecules that are entwined and regulate this process, among which cytokines are known to be expressed and are still least explored. Two of such cytokines we have used for our study are IL-1β and its natural antagonist IL-1ra to understand the functional dynamics of cytokines involved in the hatching process. Using hamster, an intriguing experimental model for hatching behavior, we have shown the mRNA (qPCR) and protein (ICC) expression of IL-1β, IL-1ra and IL-1 receptor type 1 throughout all the stages of morula, blastocyst and hatched blastocyst. Post-asserting the expression, the functional role is shown by supplementation studies, where IL-1β supplementation showed enhancement in hatching level (IL-1β treated: 84.1 ± 4.2% vs control: 63.7 ± 3.1 %, N=11), further confirmed by the diminishing effect of IL-1ra on hatching rate (IL-1ra treated: 27.5 ± 11.1% vs control: 67.9 ± 3.1%). The exogenous supplementation of IL-1ra decreased the survival rate of embryos and affected the viability in dose-dependent manner, establishing the importance of IL-1β in blastocyst cell survival. Previously, the cathepsin L and B were established as the proteases that were involved in the hamster hatching process. The inducing effect of IL-1β was correlated with enhanced mRNA level, as analyzed by qPCR, for both CAT L (by 1.9 ± 0.5 fold) and CAT B (by 3.5 ± 0.1) fold which was diminished in presence of IL-1ra (Cat L by 88 percent and Cat B by 94 percent. Moreover, using a specific fluorescent substrate-based assay kit, the enzymatic activity of these proteases was found to be increased in presence of IL-1β (Cat L by 2.1 ± 0.1 fold and CAT B by 2.3 ± 0.7 fold) and was curtailed in presence of IL-1ra. These observations provide functional insights with respect to the involvement of cytokines in the hatching process. This has implications in understanding the hatching biology and improving the embryo development quality in IVF clinics.Keywords: Blastocyst, Cytokines, Hatching, Interleukin
Procedia PDF Downloads 144234 DNA Methylation Changes in Response to Ocean Acidification at the Time of Larval Metamorphosis in the Edible Oyster, Crassostrea hongkongensis
Authors: Yong-Kian Lim, Khan Cheung, Xin Dang, Steven Roberts, Xiaotong Wang, Vengatesen Thiyagarajan
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Unprecedented rate of increased CO₂ level in the ocean and the subsequent changes in carbonate system including decreased pH, known as ocean acidification (OA), is predicted to disrupt not only the calcification process but also several other physiological and developmental processes in a variety of marine organisms, including edible oysters. Nonetheless, not all species are vulnerable to those OA threats, e.g., some species may be able to cope with OA stress using environmentally induced modifications on gene and protein expressions. For example, external environmental stressors, including OA, can influence the addition and removal of methyl groups through epigenetic modification (e.g., DNA methylation) process to turn gene expression “on or off” as part of a rapid adaptive mechanism to cope with OA. In this study, the above hypothesis was tested through testing the effect of OA, using decreased pH 7.4 as a proxy, on the DNA methylation pattern of an endemic and a commercially important estuary oyster species, Crassostrea hongkongensis, at the time of larval habitat selection and metamorphosis. Larval growth rate did not differ between control pH 8.1 and treatment pH 7.4. The metamorphosis rate of the pediveliger larvae was higher at pH 7.4 than those in control pH 8.1; however, over one-third of the larvae raised at pH 7.4 failed to attach to an optimal substrate as defined by biofilm presence. During larval development, a total of 130 genes were differentially methylated across the two treatments. The differential methylation in the larval genes may have partially accounted for the higher metamorphosis success rate under decreased pH 7.4 but with poor substratum selection ability. Differentially methylated loci were concentrated in the exon regions and appear to be associated with cytoskeletal and signal transduction, oxidative stress, metabolic processes, and larval metamorphosis, which implies the high potential of C. hongkongensis larvae to acclimate and adapt through non-genetic ways to OA threats within a single generation.Keywords: adaptive plasticity, DNA methylation, larval metamorphosis, ocean acidification
Procedia PDF Downloads 139233 Effect of Electropolymerization Method in the Charge Transfer Properties and Photoactivity of Polyaniline Photoelectrodes
Authors: Alberto Enrique Molina Lozano, María Teresa Cortés Montañez
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Polyaniline (PANI) photoelectrodes were electrochemically synthesized through electrodeposition employing three techniques: chronoamperometry (CA), cyclic voltammetry (CV), and potential pulse (PP) methods. The substrate used for electrodeposition was a fluorine-doped tin oxide (FTO) glass with dimensions of 2.5 cm x 1.3 cm. Subsequently, structural and optical characterization was conducted utilizing Fourier-transform infrared (FTIR) spectroscopy and UV-visible (UV-vis) spectroscopy, respectively. The FTIR analysis revealed variations in the molar ratio of benzenoid to quinonoid rings within the PANI polymer matrix, indicative of differing oxidation states arising from the distinct electropolymerization methodologies employed. In the optical characterization, differences in the energy band gap (Eg) values and positions of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) were observed, attributable to variations in doping levels and structural irregularities introduced during the electropolymerization procedures. To assess the charge transfer properties of the PANI photoelectrodes, electrochemical impedance spectroscopy (EIS) experiments were carried out within a 0.1 M sodium sulfate (Na₂SO₄) electrolyte. The results displayed a substantial decrease in charge transfer resistance with the PANI coatings compared to uncoated substrates, with PANI obtained through cyclic voltammetry (CV) presenting the lowest charge transfer resistance, contrasting PANI obtained via chronoamperometry (CA) and potential pulses (PP). Subsequently, the photoactive response of the PANI photoelectrodes was measured through linear sweep voltammetry (LSV) and chronoamperometry. The photoelectrochemical measurements revealed a discernible photoactivity in all PANI-coated electrodes. However, PANI electropolymerized through CV displayed the highest photocurrent. Interestingly, PANI derived from chronoamperometry (CA) exhibited the highest degree of stable photocurrent over an extended temporal interval.Keywords: PANI, photocurrent, photoresponse, charge separation, recombination
Procedia PDF Downloads 65232 Compact 3-D Co-Planar Waveguide Fed Dual-Port Ultrawideband-Multiple-Input and Multiple-Output Antenna with WLAN Band-Notched Characteristics
Authors: Asim Quddus
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A miniaturized three dimensional co-planar waveguide (CPW) two-port MIMO antenna, exhibiting high isolation and WLAN band-notched characteristics is presented in this paper for ultrawideband (UWB) communication applications. The microstrip patch antenna operates as a single UWB antenna element. The proposed design is a cuboid-shaped structure having compact size of 35 x 27 x 45 mm³. Radiating as well as decoupling structure is placed around cuboidal polystyrene sheet. The radiators are 27 mm apart, placed Face-to-Face in vertical direction. Decoupling structure is placed on the side walls of polystyrene. The proposed antenna consists of an oval shaped radiating patch. A rectangular structure with fillet edges is placed on ground plan to enhance the bandwidth. The proposed antenna exhibits a good impedance match (S11 ≤ -10 dB) over frequency band of 2 GHz – 10.6 GHz. A circular slotted structure is employed as a decoupling structure on substrate, and it is placed on the side walls of polystyrene to enhance the isolation between antenna elements. Moreover, to achieve immunity from WLAN band distortion, a modified, inverted crescent shaped slotted structure is etched on radiating patches to achieve band-rejection characteristics at WLAN frequency band 4.8 GHz – 5.2 GHz. The suggested decoupling structure provides isolation better than 15 dB over the desired UWB spectrum. The envelope correlation coefficient (ECC) and gain for the MIMO antenna are analyzed as well. Finite Element Method (FEM) simulations are carried out in Ansys High Frequency Structural Simulator (HFSS) for the proposed design. The antenna is realized on a Rogers RT/duroid 5880 with thickness 1 mm, relative permittivity ɛr = 2.2. The proposed antenna achieves a stable omni-directional radiation patterns as well, while providing rejection at desired WLAN band. The S-parameters as well as MIMO parameters like ECC are analyzed and the results show conclusively that the design is suitable for portable MIMO-UWB applications.Keywords: 3-D antenna, band-notch, MIMO, UWB
Procedia PDF Downloads 296231 Development of a Microfluidic Device for Low-Volume Sample Lysis
Authors: Abbas Ali Husseini, Ali Mohammad Yazdani, Fatemeh Ghadiri, Alper Şişman
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We developed a microchip device that uses surface acoustic waves for rapid lysis of low level of cell samples. The device incorporates sharp-edge glass microparticles for improved performance. We optimized the lysis conditions for high efficiency and evaluated the device's feasibility for point-of-care applications. The microchip contains a 13-finger pair interdigital transducer with a 30-degree focused angle. It generates high-intensity acoustic beams that converge 6 mm away. The microchip operates at a frequency of 16 MHz, exciting Rayleigh waves with a 250 µm wavelength on the LiNbO3 substrate. Cell lysis occurs when Candida albicans cells and glass particles are placed within the focal area. The high-intensity surface acoustic waves induce centrifugal forces on the cells and glass particles, resulting in cell lysis through lateral forces from the sharp-edge glass particles. We conducted 42 pilot cell lysis experiments to optimize the surface acoustic wave-induced streaming. We varied electrical power, droplet volume, glass particle size, concentration, and lysis time. A regression machine-learning model determined the impact of each parameter on lysis efficiency. Based on these findings, we predicted optimal conditions: electrical signal of 2.5 W, sample volume of 20 µl, glass particle size below 10 µm, concentration of 0.2 µg, and a 5-minute lysis period. Downstream analysis successfully amplified a DNA target fragment directly from the lysate. The study presents an efficient microchip-based cell lysis method employing acoustic streaming and microparticle collisions within microdroplets. Integration of a surface acoustic wave-based lysis chip with an isothermal amplification method enables swift point-of-care applications.Keywords: cell lysis, surface acoustic wave, micro-glass particle, droplet
Procedia PDF Downloads 79230 Plasma Treatment in Conjunction with EGM-2 Medium Can Enhance Endothelial and Osteogenic Marker Expressions of Bone Marrow MSCs
Authors: Chih-Hsin Lin, Shyh-Yuan Lee, Yuan-Min Lin
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For many tissue engineering applications, an important goal is to create functional tissues in-vitro, and such tissues to be viable, they have to be vascularized. Endothelial cells (EC) and endothelial progenitor cells (EPC) are promising candidates for vascularization. However, both of them have limited expansion capacity and autologous cells currently do not exist for either ECs or EPCs. Therefore, we use bone marrow mesenchymal stem cells (MSC) as a source material for ECs. Growth supplements are commonly used to induce MSC differentiation, and further improvements in differentiation conditions can be made by modifying the cell's growth environment. An example is pre-treatment of the growth dish with gas plasma, in order to modify the surface functional groups of the material that the cells are seeded on. In this work, we compare the effects of different gas plasmas on the growth and differentiation of MSCs. We treat the dish with different plasmas (CO2, N2, and O2) and then induce MSC differentiation with endothelial growth medium-2 (EGM-2). We find that EGM-2 by itself upregulates EC marker CD31 mRNA expression, but not VEGFR2, CD34, or vWF. However, these additional EC marker expressions were increased for cells seeded on plasma treated substrates. Specifically, for EC markers, we found that N2 plasma treatment upregulated CD31 and VEGFR-2 mRNA expressions; CO2 plasma treatment upregulated CD34 and vWF mRNA expressions. The osteogenic markers ALP and osteopontin mRNA expressions were markedly enhanced on all plasma-treated dishes. We also found that plasma treatment in conjunction with EGM-2 growth medium can enhance MSCs differentiation into endothelial-like cells and osteogenic-like cells. Our work shows that the effect of the growth medium (EGM-2) on MSCs differentiation is influenced by the plasma modified surface chemistry of the substrate. In conclusion, plasma surface modification can enhance EGM-2 effectiveness and induced both endothelial and osteogenic differentiation. Our findings provide a method to enhance EGM-2 based cell differentiation, with consequences for tissue engineering and stem cell biology applications.Keywords: endothelial differentiation, EGM-2, osteogenesis, plasma treatment, surface modification
Procedia PDF Downloads 329229 Electrochemical Bioassay for Haptoglobin Quantification: Application in Bovine Mastitis Diagnosis
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 173228 Fermentation of Pretreated Herbaceous Cellulosic Wastes to Ethanol by Anaerobic Cellulolytic and Saccharolytic Thermophilic Clostridia
Authors: Lali Kutateladze, Tamar Urushadze, Tamar Dudauri, Besarion Metreveli, Nino Zakariashvili, Izolda Khokhashvili, Maya Jobava
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Lignocellulosic waste streams from agriculture, paper and wood industry are renewable, plentiful and low-cost raw materials that can be used for large-scale production of liquid and gaseous biofuels. As opposed to prevailing multi-stage biotechnological processes developed for bioconversion of cellulosic substrates to ethanol where high-cost cellulase preparations are used, Consolidated Bioprocessing (CBP) offers to accomplish cellulose and xylan hydrolysis followed by fermentation of both C6 and C5 sugars to ethanol in a single-stage process. Syntrophic microbial consortium comprising of anaerobic, thermophilic, cellulolytic, and saccharolytic bacteria in the genus Clostridia with improved ethanol productivity and high tolerance to fermentation end-products had been proposed for achieving CBP. 65 new strains of anaerobic thermophilic cellulolytic and saccharolytic Clostridia were isolated from different wetlands and hot springs in Georgia. Using new isolates, fermentation of mechanically pretreated wheat straw and corn stalks was done under oxygen-free nitrogen environment in thermophilic conditions (T=550C) and pH 7.1. Process duration was 120 hours. Liquid and gaseous products of fermentation were analyzed on a daily basis using Perkin-Elmer gas chromatographs with flame ionization and thermal detectors. Residual cellulose, xylan, xylose, and glucose were determined using standard methods. Cellulolytic and saccharolytic bacteria strains degraded mechanically pretreated herbaceous cellulosic wastes and fermented glucose and xylose to ethanol, acetic acid and gaseous products like hydrogen and CO2. Specifically, maximum yield of ethanol was reached at 96 h of fermentation and varied between 2.9 – 3.2 g/ 10 g of substrate. The content of acetic acid didn’t exceed 0.35 g/l. Other volatile fatty acids were detected in trace quantities.Keywords: anaerobic bacteria, cellulosic wastes, Clostridia sp, ethanol
Procedia PDF Downloads 294227 X-Ray Crystallographic Studies on BPSL2418 from Burkholderia pseudomallei
Authors: Mona Alharbi
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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis
Procedia PDF Downloads 248226 Utilization of Agro-wastes for Biotechnological Production of Edible Mushroom
Authors: Salami Abiodun Olusola, Bankole Faith Ayobami
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Agro-wastes are wastes produced from various agricultural activities and include manures, corncob, plant stalks, hulls, leaves, sugarcane bagasse, oil-palm spadix, and rice bran. In farming situation, the agro-waste is often useless and, thus, discarded. Huge quantities of waste resources generated from Nigerian agriculture could be converted to more useful forms of energy, which could contribute to the country’s primary energy needs and reduce problems associated with waste management. Accumulation of agro-wastes may cause health, safety, and environmental concern. However, biotechnological use of agro-waste could enhance food security through its bioconversion to useful renewable energy. Mushrooms are saprophytes which feed by secreting extracellular enzymes, digesting food externally, and absorb the nutrients in net-like hyphae. Therefore, mushrooms could be exploited for bioconversion of the cheap and numerous agro-wastes for providing nutritious food for animals, human and carbon recycling. The study investigated the bioconversion potentials of Pleurotus florida on agro-wastes using a simple and cost-effective biotechnological method. Four agro-wastes; corncobs, oil-palm spadix, corn straw, and sawdust, were composted and used as substrates while the biological efficiency (BE) and the nutritional composition of P. florida grown on the substrates were determined. Pleurotus florida contained 26.28-29.91% protein, 86.90-89.60% moisture, 0.48-0.91% fat, 19.64-22.82% fibre, 31.37-38.17% carbohydrate and 5.18-6.39% ash. The mineral contents ranged from 342-410 mg/100g Calcium, 1009-1133 mg/100g Phosphorus, 17-21 mg/100g Iron, 277-359 mg/100g Sodium, and 2088-2281 mg/100g Potassium. The highest yield and BE were obtained on corncobs (110 g, 55%), followed by oil-palm spadix (76.05 g, 38%), while the least BE was recorded on corn straw substrate (63.12 g, 31.56%). Utilization of the composted substrates yielded nutritional and edible mushrooms. The study presents biotechnological procedure for bioconversion of agro-wastes to edible and nutritious mushroom for efficient agro-wastes’ management, utilization, and recycling.Keywords: agrowaste, bioconversion, biotechnology, utilization, recycling
Procedia PDF Downloads 77225 Carbon Nanofibers as the Favorite Conducting Additive for Mn₃O₄ Catalysts for Oxygen Reactions in Rechargeable Zinc-Air Battery
Authors: Augustus K. Lebechi, Kenneth I. Ozoemena
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Rechargeable zinc-air batteries (RZABs) have been described as one of the most viable next-generation ‘beyond-the-lithium-ion’ battery technologies with great potential for renewable energy storage. It is safe, with a high specific energy density (1086 Wh/kg), environmentally benign, and low-cost, especially in resource-limited African countries. For widespread commercialization, the sluggish oxygen reaction kinetics pose a major challenge that impedes the reversibility of the system. Hence, there is a need for low-cost and highly active bifunctional electrocatalysts. Manganese oxide catalysts on carbon conducting additives remain the best couple for the realization of such low-cost RZABs. In this work, hausmannite Mn₃O₄ nanoparticles were synthesized through the annealing method from commercial electrolytic manganese dioxide (EMD), multi-walled carbon nanotubes (MWCNTs) were synthesized via the chemical vapor deposition (CVD) method and carbon nanofibers (CNFs) were synthesized via the electrospinning process with subsequent carbonization. Both Mn₃O₄ catalysts and the carbon conducting additives (MWCNT and CNF) were thoroughly characterized using X-ray powder diffraction spectroscopy (XRD), scanning electron microscopy (SEM), thermogravimetry analysis (TGA) and X-ray photoelectron spectroscopy (XPS). Composite electrocatalysts (Mn₃O₄/CNT and Mn₃O₄/CNF) were investigated for oxygen evolution reaction (OER) and oxygen reduction reaction (ORR) in an alkaline medium. Using the established electrocatalytic modalities for evaluating the electrocatalytic performance of materials (including double layer, electrochemical active surface area, roughness factor, specific current density, and catalytic stability), CNFs proved to be the most efficient conducting additive material for the Mn₃O₄ catalyst. From the DFT calculations, the higher performance of the CNFs over the MWCNTs is related to the ability of the CNFs to allow for a more favorable distribution of the d-electrons of the manganese (Mn) and enhanced synergistic effect with Mn₃O₄ for weaker adsorption energies of the oxygen intermediates (O*, OH* and OOH*). In a proof-of-concept, Mn₃O₄/CNF was investigated as the air cathode for rechargeable zinc-air battery (RZAB) in a micro-3D-printed cell configuration. The RZAB showed good performance in terms of open circuit voltage (1.77 V), maximum power density (177.5 mW cm-2), areal-discharge energy and cycling stability comparable to Pt/C (20 wt%) + IrO2. The findings here provide fresh physicochemical perspectives on the future design and utility of CNFs for developing manganese-based RZABs.Keywords: bifunctional electrocatalyst, oxygen evolution reaction, oxygen reduction reactions, rechargeable zinc-air batteries.
Procedia PDF Downloads 64224 Cybernetic Model-Based Optimization of a Fed-Batch Process for High Cell Density Cultivation of E. Coli In Shake Flasks
Authors: Snehal D. Ganjave, Hardik Dodia, Avinash V. Sunder, Swati Madhu, Pramod P. Wangikar
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Batch cultivation of recombinant bacteria in shake flasks results in low cell density due to nutrient depletion. Previous protocols on high cell density cultivation in shake flasks have relied mainly on controlled release mechanisms and extended cultivation protocols. In the present work, we report an optimized fed-batch process for high cell density cultivation of recombinant E. coli BL21(DE3) for protein production. A cybernetic model-based, multi-objective optimization strategy was implemented to obtain the optimum operating variables to achieve maximum biomass and minimized substrate feed rate. A syringe pump was used to feed a mixture of glycerol and yeast extract into the shake flask. Preliminary experiments were conducted with online monitoring of dissolved oxygen (DO) and offline measurements of biomass and glycerol to estimate the model parameters. Multi-objective optimization was performed to obtain the pareto front surface. The selected optimized recipe was tested for a range of proteins that show different extent soluble expression in E. coli. These included eYFP and LkADH, which are largely expressed in soluble fractions, CbFDH and GcanADH , which are partially soluble, and human PDGF, which forms inclusion bodies. The biomass concentrations achieved in 24 h were in the range 19.9-21.5 g/L, while the model predicted value was 19.44 g/L. The process was successfully reproduced in a standard laboratory shake flask without online monitoring of DO and pH. The optimized fed-batch process showed significant improvement in both the biomass and protein production of the tested recombinant proteins compared to batch cultivation. The proposed process will have significant implications in the routine cultivation of E. coli for various applications.Keywords: cybernetic model, E. coli, high cell density cultivation, multi-objective optimization
Procedia PDF Downloads 257223 Thermal Stability and Electrical Conductivity of Ca₅Mg₄₋ₓMₓ(VO₄)₆ (0 ≤ x ≤ 4) where M = Zn, Ni Measured by Impedance Spectroscopy
Authors: Anna S. Tolkacheva, Sergey N. Shkerin, Kirill G. Zemlyanoi, Olga G. Reznitskikh, Pavel D. Khavlyuk
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Calcium oxovanadates with garnet related structure are multifunctional oxides in various fields like photoluminescence, microwave dielectrics, and magneto-dielectrics. For example, vanadate garnets are self-luminescent compounds. They attract attention as RE-free broadband excitation and emission phosphors and are candidate materials for UV-based white light-emitting diodes (WLEDs). Ca₅M₄(VO₄)₆ (M = Mg, Zn, Co, Ni, Mn) compounds are also considered promising for application in microwave devices as substrate materials. However, the relation between their structure, composition and physical/chemical properties remains unclear. Given the above-listed observations, goals of this study are to synthesise Ca₅M₄(VO₄)₆ (M = Mg, Zn, Ni) and to study their thermal and electrical properties. Solid solutions Ca₅Mg₄₋ₓMₓ(VO₄)₆ (0 ≤ x ≤ 4) where M is Zn and Ni have been synthesized by sol-gel method. The single-phase character of the final products was checked by powder X-ray diffraction on a Rigaku D/MAX-2200 X-ray diffractometer using Cu Kα radiation in the 2θ range from 15° to 70°. The dependence of thermal properties on chemical composition of solid solutions was studied using simultaneous thermal analyses (DSC and TG). Thermal analyses were conducted in a Netzch simultaneous analyser STA 449C Jupiter, in Ar atmosphere, in temperature range from 25 to 1100°C heat rate was 10 K·min⁻¹. Coefficients of thermal expansion (CTE) were obtained by dilatometry measurements in air up to 800°C using a Netzsch 402PC dilatometer; heat rate was 1 K·min⁻¹. Impedance spectra were obtained via the two-probe technique with an impedance meter Parstat 2273 in air up to 700°C with the variation of pH₂O from 0.04 to 3.35 kPa. Cation deficiency in Ca and Mg sublattice under the substitution of MgO with ZnO up to 1/6 was observed using Rietveld refinement of the crystal structure. Melting point was found to decrease with x changing from 0 to 4 in Ca₅Mg₄₋ₓMₓ(VO₄)₆ where M is Zn and Ni. It was observed that electrical conductivity does not depend on air humidity. The reported study was funded by the RFBR Grant No. 17–03–01280. Sample attestation was carried out in the Shared Access Centers at the IHTE UB RAS.Keywords: garnet structure, electrical conductivity, thermal expansion, thermal properties
Procedia PDF Downloads 155222 Distribution Patterns of the Renieramycin-M-Producing Blue Sponge, Xestospongia sp. (De Laubenfels, 1932) (Phylum: Porifera, Class: Demospongiae) in Puerto Galera, Oriental Mindoro, Philippines
Authors: Geminne Manzano, Clairecynth Yu, Lilibeth Salvador-Reyes, Viviene Santiago, Porfirio AliñO
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The distribution and abundance patterns of many marine sessile organisms such as sponges vary among and within reefs. Determining the factors affecting its distribution is essential especially for organisms that produce secondary metabolites with pharmaceutical importance. In this study, the small-scale distribution patterns of the Philippine blue sponge, Xestospongia sp. in relation to some ecological factors were examined. The relationship between the renieramycin-M production and their benthic attributes were also determined. Ecological surveys were conducted on two stations with varying depth and exposure located in Oriental Mindoro, Philippines. Three 30 by 6m belt transect were used to assess the sponge abundance at each station. The substratum of the sponges was also characterized. Fish visual census observations were also taken together with the photo transect methods benthic surveys. Sponge samples were also collected for the extraction of Renieramycin-M and for further chemical analysis. Varying distribution patterns were observed to be attributed to the combination of different ecological and environmental factors. The amount of Renieramycin-production also varied in each station. The common substratum for blue sponges includes hard and soft corals, as well as, dead coral with algal patches. Blue sponges from exposed habitat frequently grow associated with massive and branching corals, Porites sp., while the most frequent substrate found on sheltered habitats is the coral Pavona sp. Exploring the influence of ecological and environmental parameters on the abundance and distribution of sponge assemblages provide ecological insights and their potential applications to pharmaceutical studies. The results of this study provide further impetus in pursuing studies into patterns and processes of the Philippine blue sponge, Xestospongia sp. distribution in relation to the chemical ecology of its secondary metabolites.Keywords: distribution patterns, Porifera, Renieramycin-M, sponge assemblages, Xestospongia sp.
Procedia PDF Downloads 266