Search results for: vegetable substrate
Commenced in January 2007
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Edition: International
Paper Count: 1384

Search results for: vegetable substrate

244 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

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243 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

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242 Food Safety in Wine: Removal of Ochratoxin a in Contaminated White Wine Using Commercial Fining Agents

Authors: Antònio Inês, Davide Silva, Filipa Carvalho, Luís Filipe-Riberiro, Fernando M. Nunes, Luís Abrunhosa, Fernanda Cosme

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The presence of mycotoxins in foodstuff is a matter of concern for food safety. Mycotoxins are toxic secondary metabolites produced by certain molds, being ochratoxin A (OTA) one of the most relevant. Wines can also be contaminated with these toxicants. Several authors have demonstrated the presence of mycotoxins in wine, especially ochratoxin A. Its chemical structure is a dihydro-isocoumarin connected at the 7-carboxy group to a molecule of L-β-phenylalanine via an amide bond. As these toxicants can never be completely removed from the food chain, many countries have defined levels in food in order to attend health concerns. OTA contamination of wines might be a risk to consumer health, thus requiring treatments to achieve acceptable standards for human consumption. The maximum acceptable level of OTA in wines is 2.0 μg/kg according to the Commission regulation No. 1881/2006. Therefore, the aim of this work was to reduce OTA to safer levels using different fining agents, as well as their impact on white wine physicochemical characteristics. To evaluate their efficiency, 11 commercial fining agents (mineral, synthetic, animal and vegetable proteins) were used to get new approaches on OTA removal from white wine. Trials (including a control without addition of a fining agent) were performed in white wine artificially supplemented with OTA (10 µg/L). OTA analyses were performed after wine fining. Wine was centrifuged at 4000 rpm for 10 min and 1 mL of the supernatant was collected and added of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v). Also, the solid fractions obtained after fining, were centrifuged (4000 rpm, 15 min), the resulting supernatant discarded, and the pellet extracted with 1 mL of the above solution and 1 mL of H2O. OTA analysis was performed by HPLC with fluorescence detection. The most effective fining agent in removing OTA (80%) from white wine was a commercial formulation that contains gelatin, bentonite and activated carbon. Removals between 10-30% were obtained with potassium caseinate, yeast cell walls and pea protein. With bentonites, carboxymethylcellulose, polyvinylpolypyrrolidone and chitosan no considerable OTA removal was verified. Following, the effectiveness of seven commercial activated carbons was also evaluated and compared with the commercial formulation that contains gelatin, bentonite and activated carbon. The different activated carbons were applied at the concentration recommended by the manufacturer in order to evaluate their efficiency in reducing OTA levels. Trial and OTA analysis were performed as explained previously. The results showed that in white wine all activated carbons except one reduced 100% of OTA. The commercial formulation that contains gelatin, bentonite and activated carbon reduced only 73% of OTA concentration. These results may provide useful information for winemakers, namely for the selection of the most appropriate oenological product for OTA removal, reducing wine toxicity and simultaneously enhancing food safety and wine quality.

Keywords: wine, ota removal, food safety, fining

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241 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

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240 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

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239 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

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238 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

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237 Functional Ingredients from Potato By-Products: Innovative Biocatalytic Processes

Authors: Salwa Karboune, Amanda Waglay

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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.

Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides

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236 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

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235 Anti-Hyperglycemic Effects and Chemical Analysis of Allium sativum Bulbs Growing in Sudan

Authors: Ikram Mohamed Eltayeb Elsiddig, Yacouba Amina Djamila, Amna El Hassan Hamad

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Hyperglycemia and diabetes have been treated with several medicinal plants for a long time, meanwhile reduce associated side effects than the synthetic ones. Therefore, the search for more effective and safer anti-diabetic agents derived from plants has become an interest area of active research. A. sativum, belonging to the Liliaceae family is well known for its medicinal uses in African traditional medicine, it used for treating of many human diseases mainly diabetes, high cholesterol, and high blood pressure. The present study was carried out to investigate the anti-hyperglycemic effect of the extracts of A. sativum bulb growing in Sudan on glucose-loaded Wistar albino rats. A. sativum bulbs were collected from local vegetable market at Khourtoum/ Sudan in a fresh form, identified and authenticated by taxonomist, then dried, and extracted with solvents of increasing polarity: petroleum ether, chloroform, ethyl acetate and methanol by using Soxhlet apparatus. The effect of the extracts on glucose uptake was evaluated by using the isolated rats hemidiaphgrams after loading the fasting rats with glucose, and the anti-hyperglycemic effect was investigated on glucose-loaded Wistar albino rats. Their effects were compared to control rats administered with the vehicle and to a standard group administered with Metformin standard drug. The most active extract was analyzed chemically using GC-MS analysis compared to NIST library. The results showed significant anti-diabetic effect of extracts of A. sativum bulb growing in Sudan. Addition to the hypoglycemic activity of A. sativum extracts was found to be decreased with increase in the polarity of the extraction solvent; this may explain the less polarity of substance responsible for the activity and their concentration decreased with polarity increase. The petroleum ether extract possess anti-hyperglycemic activity more significant than the other extracts and the Metformin standard drug with p-value 0.000** of 400mg/kg at 1 hour, 2 hour and four hour; and p-value 0.019*, 0.015* and 0.010* of 200mg/kg at 1 hour, 2 hour and four hour respectively. The GC-MS analysis of petroleum ether extract, with highest anti -diabetes activity showed the presence of Methyl linolate (42.75%), Hexadecanoic acid, methyl ester (10.54%), Methyl α-linolenate (8.36%), Dotriacontane (6.83), Tetrapentacontane (6.33), Methyl 18-methylnonadecanoate (4.8), Phenol,2,2’-methylenebis[6-(1,1-dimethylethyl)-4-methyl] (3.25), Methyl 20-methyl-heneicosanoate (2.70), Pentatriacontane (2.13) and many other minor compounds. The most of these compounds are well known for their anti-diabetic activity. The study concluded that A. sativum bulbs extracts were found to enhanced the reuptake of glucose in the isolated rat hemidiaphragm and have antihyperglycemic effect when evaluated on glucose-loaded albino rats with petroleum ether extract activity more significant than the Metformin standard drug.

Keywords: Allium, anti-hyperglycemic, bulbs, sativum

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234 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

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233 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

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232 Elderly in Sub Saharan Africa

Authors: Obinna Benedict Duru

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This study focuses on the elderly and the challenges that confront them. The elderly are that particular segment of our population who by virtue of the aging process have attained the stage in most cases where they are confronted with the challenges of economic dependency and social marginality. These challenges are as a result of the physical and biological decline occasioned by social myths and realities which portray the elderly as a dependent population whose members could not and should not work and who need social assistance that the younger population is obliged to provide. From the moment of birth to the moment of death, our bodies are constantly changing. We are all enmeshed in the process of growing old, a transition from youthfulness to elderliness. In youth-oriented modern societies like ours, we tend to attach positive importance and significance to the biological changes that occur early in life and define later physical changes in negative terms. Children growing up and young adults receive more attention, greater responsibilities and more legal rights to reward them on their way. But few people are congratulated on getting old. We commiserate with people who are getting old and make jokes about their supposedly physical, mental and biological decline. Wrinkles, loss of weight and vitality are all parts of the aging process. In almost all parts of the world, earlier researches have shown that about fifty percent of the elderly who suffer from stroke, arthritis, senility and other age related diseases are the disengaged and neglected elderly. Rapid technological changes render the knowledge and skills of the elderly obsolete; education is geared toward the young and the generational competition for jobs leads to pressures on the elderly to retire. Control of initial resources are shifted to the middle-aged and older workers are pushed into positions of economic dependency. This study therefore, among other things tend to discover how some government policies have affected the elderly particularly in Africa. To discover the prospects and possibilities of the elderly for a better living. To make a comparison of the advances in healthcare giving made in the advanced western societies to the practice in Sub Saharan Africa etc. The hypotheses of this study include: that the elderly in Sub Saharan Africa are more vulnerable than their counterparts in Europe and America. The elderly are more prone to social isolation, and that the elderly are mostly affected by age-related sickness etc. With a survey method as the research design, and sample size of about 500 respondents,probability sampling technique was used. Data which were analyzed using chi-square and tables were collected through primary and secondary sources. The findings made include: that the elderly suffer pains of old age especially when disengaged from work or social activity. That loss of income condemn the elderly to a life of vegetable existence, and that those who do not have other means of re-integration usually see old age with regret and despair. It is therefore, recommended among other things that social welfare scheme and the process of re-integration at old age be introduced for the non pensionable elderly in Africa.

Keywords: elderly, social isolation, dependency, re-integration

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231 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

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230 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 247
229 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 218
228 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 42
227 Intermodal Strategies for Redistribution of Agrifood Products in the EU: The Case of Vegetable Supply Chain from Southeast of Spain

Authors: Juan C. Pérez-Mesa, Emilio Galdeano-Gómez, Jerónimo De Burgos-Jiménez, José F. Bienvenido-Bárcena, José F. Jiménez-Guerrero

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Environmental cost and transport congestion on roads resulting from product distribution in Europe have to lead to the creation of various programs and studies seeking to reduce these negative impacts. In this regard, apart from other institutions, the European Commission (EC) has designed plans in recent years promoting a more sustainable transportation model in an attempt to ultimately shift traffic from the road to the sea by using intermodality to achieve a model rebalancing. This issue proves especially relevant in supply chains from peripheral areas of the continent, where the supply of certain agrifood products is high. In such cases, the most difficult challenge is managing perishable goods. This study focuses on new approaches that strengthen the modal shift, as well as the reduction of externalities. This problem is analyzed by attempting to promote intermodal system (truck and short sea shipping) for transport, taking as point of reference highly perishable products (vegetables) exported from southeast Spain, which is the leading supplier to Europe. Methodologically, this paper seeks to contribute to the literature by proposing a different and complementary approach to establish a comparison between intermodal and the “only road” alternative. For this purpose, the multicriteria decision is utilized in a p-median model (P-M) adapted to the transport of perishables and to a means of shipping selection problem, which must consider different variables: transit cost, including externalities, time, and frequency (including agile response time). This scheme avoids bias in decision-making processes. By observing the results, it can be seen that the influence of the externalities as drivers of the modal shift is reduced when transit time is introduced as a decision variable. These findings confirm that the general strategies, those of the EC, based on environmental benefits lose their capacity for implementation when they are applied to complex circumstances. In general, the different estimations reveal that, in the case of perishables, intermodality would be a secondary and viable option only for very specific destinations (for example, Hamburg and nearby locations, the area of influence of London, Paris, and the Netherlands). Based on this framework, the general outlook on this subject should be modified. Perhaps the government should promote specific business strategies based on new trends in the supply chain, not only on the reduction of externalities, and find new approaches that strengthen the modal shift. A possible option is to redefine ports, conceptualizing them as digitalized redistribution and coordination centers and not only as areas of cargo exchange.

Keywords: environmental externalities, intermodal transport, perishable food, transit time

Procedia PDF Downloads 67
226 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 201
225 Relative Expression and Detection of MUB Adhesion Domains and Plantaricin-Like Bacteriocin among Probiotic Lactobacillus plantarum-Group Strains Isolated from Fermented Foods

Authors: Sundru Manjulata Devi, Prakash M. Halami

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The immemorial use of fermented foods from vegetables, dairy and other biological sources are of great demand in India because of their health benefits. However, the diversity of Lactobacillus plantarum group (LPG) of vegetable origin has not been revealed yet, particularly with reference to their probiotic functionalities. In the present study, the different species of probiotic Lactobacillus plantarum group (LPG) i.e., L. plantarum subsp. plantarum MTCC 5422 (from fermented cereals), L. plantarum subsp. argentoratensis FG16 (from fermented bamboo shoot) and L. paraplantarum MTCC 9483 (from fermented gundruk) (as characterized by multiplex recA PCR assay) were considered to investigate their relative expression of MUB domains of mub gene (mucin binding protein) by Real time PCR. Initially, the allelic variation in the mub gene was assessed and found to encode three different variants (Type I, II and III). All the three types had 8, 9 and 10 MUB domains respectively (as analysed by Pfam database) and were found to be responsible for adhesion of bacteria to the host intestinal epithelial cells. These domains either get inserted or deleted during speciation or evolutionary events and lead to divergence. The reverse transcriptase qPCR analysis with mubLPF1+R1 primer pair supported variation in amplicon sizes with 300, 500 and 700 bp among different LPG strains. The relative expression of these MUB domains significantly unregulated in the presence of 1% mucin in overnight grown cultures. Simultaneously, the mub gene expressed efficiently by 7 fold in the culture L. paraplantarum MTCC 9483 with 10 MUB domains. An increase in the expression levels for L. plantarum subsp. plantarum MTCC 5422 and L. plantarum subsp. argentoratensis FG16 (MCC 2974) with 9 and 8 repetitive domains was around 4 and 2 fold, respectively. The detection and expression of an integrase (int) gene in the upstream region of mub gene reveals the excision and integration of these repetitive domains. Concurrently, an in vitro adhesion assay to mucin and exclusion of pathogens (such as Listeria monocytogenes and Micrococcus leuteus) was investigated and observed that the L. paraplantarum MTCC 9483 with more adhesion domains has more ability to adhere to mucin and inhibited the growth of pathogens. The production and expression of plantaricin-like bacteriocin (plnNC8 type) in MTCC 9483 suggests the pathogen inhibition. Hence, the expression of MUB domains can act as potential biomarkers in the screening of a novel probiotic LPG strain with adherence property. The present study provides a platform for an easy, rapid, less time consuming, low-cost methodology for the detection of potential probiotic bacteria. It was known that the traditional practices followed in the preparation of fermented bamboo shoots/gundruk/cereals of Indian foods contain different kinds of neutraceuticals for functional food and novel compounds with health promoting factors. In future, a detailed study of these food products can add more nutritive value, consumption and suitable for commercialization.

Keywords: adhesion gene, fermented foods, MUB domains, probiotics

Procedia PDF Downloads 231
224 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 130
223 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 244
222 Evaluation of the Effect of Lactose Derived Monosaccharide on Galactooligosaccharides Production by β-Galactosidase

Authors: Yenny Paola Morales Cortés, Fabián Rico Rodríguez, Juan Carlos Serrato Bermúdez, Carlos Arturo Martínez Riascos

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Numerous benefits of galactooligosaccharides (GOS) as prebiotics have motivated the study of enzymatic processes for their production. These processes have special complexities due to several factors that make difficult high productivity, such as enzyme type, reaction medium pH, substrate concentrations and presence of inhibitors, among others. In the present work the production of galactooligosaccharides (with different degrees of polymerization: two, three and four) from lactose was studied. The study considers the formulation of a mathematical model that predicts the production of GOS from lactose using the enzyme β-galactosidase. The effect of pH in the reaction was studied. For that, phosphate buffer was used and with this was evaluated three pH values (6.0.6.5 and 7.0). Thus it was observed that at pH 6.0 the enzymatic activity insignificant. On the other hand, at pH 7.0 the enzymatic activity was approximately 27 times greater than at 6.5. The last result differs from previously reported results. Therefore, pH 7.0 was chosen as working pH. Additionally, the enzyme concentration was analyzed, which allowed observing that the effect of the concentration depends on the pH and the concentration was set for the following studies in 0.272 mM. Afterwards, experiments were performed varying the lactose concentration to evaluate its effects on the process and to generate the data for the adjustment of the mathematical model parameters. The mathematical model considers the reactions of lactose hydrolysis and transgalactosylation for the production of disaccharides and trisaccharides, with their inverse reactions. The production of tetrasaccharides was negligible and, because of that, it was not included in the model. The reaction was monitored by HPLC and for the quantitative analysis of the experimental data the Matlab programming language was used, including solvers for differential equations systems integration (ode15s) and nonlinear problems optimization (fminunc). The results confirm that the transgalactosylation and hydrolysis reactions are reversible, additionally inhibition by glucose and galactose is observed on the production of GOS. In relation to the production process of galactooligosaccharides, the results show that it is necessary to have high initial concentrations of lactose considering that favors the transgalactosylation reaction, while low concentrations favor hydrolysis reactions.

Keywords: β-galactosidase, galactooligosaccharides, inhibition, lactose, Matlab, modeling

Procedia PDF Downloads 323
221 Fire Resistance of High Alumina Cement and Slag Based Ultra High Performance Fibre-Reinforced Cementitious Composites

Authors: A. Q. Sobia, M. S. Hamidah, I. Azmi, S. F. A. Rafeeqi

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Fibre-reinforced polymer (FRP) strengthened reinforced concrete (RC) structures are susceptible to intense deterioration when exposed to elevated temperatures, particularly in the incident of fire. FRP has the tendency to lose bond with the substrate due to the low glass transition temperature of epoxy; the key component of FRP matrix.  In the past few decades, various types of high performance cementitious composites (HPCC) were explored for the protection of RC structural members against elevated temperature. However, there is an inadequate information on the influence of elevated temperature on the ultra high performance fibre-reinforced cementitious composites (UHPFRCC) containing ground granulated blast furnace slag (GGBS) as a replacement of high alumina cement (HAC) in conjunction with hybrid fibres (basalt and polypropylene fibres), which could be a prospective fire resisting material for the structural components. The influence of elevated temperatures on the compressive as well as flexural strength of UHPFRCC, made of HAC-GGBS and hybrid fibres, were examined in this study. Besides control sample (without fibres), three other samples, containing 0.5%, 1% and 1.5% of basalt fibres by total weight of mix and 1 kg/m3 of polypropylene fibres, were prepared and tested. Another mix was also prepared with only 1 kg/m3 of polypropylene fibres. Each of the samples were retained at ambient temperature as well as exposed to 400, 700 and 1000 °C followed by testing after 28 and 56 days of conventional curing. Investigation of results disclosed that the use of hybrid fibres significantly helped to improve the ambient temperature compressive and flexural strength of UHPFRCC, which was found to be 80 and 14.3 MPa respectively. However, the optimum residual compressive strength was marked by UHPFRCC-CP (with polypropylene fibres only), equally after both curing days (28 and 56 days), i.e. 41%. In addition, the utmost residual flexural strength, after 28 and 56 days of curing, was marked by UHPFRCC– CP and UHPFRCC– CB2 (1 kg/m3 of PP fibres + 1% of basalt fibres) i.e. 39% and 48.5% respectively.

Keywords: fibre reinforced polymer materials (FRP), ground granulated blast furnace slag (GGBS), high-alumina cement, hybrid, fibres

Procedia PDF Downloads 261
220 Optimization of the Jatropha curcas Supply Chain as a Criteria for the Implementation of Future Collection Points in Rural Areas of Manabi-Ecuador

Authors: Boris G. German, Edward Jiménez, Sebastián Espinoza, Andrés G. Chico, Ricardo A. Narváez

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The unique flora and fauna of The Galapagos Islands has leveraged a tourism-driven growth in the islands. Nonetheless, such development is energy-intensive and requires thousands of gallons of diesel each year for thermoelectric electricity generation. The needed transport of fossil fuels from the continent has generated oil spillages and affectations to the fragile ecosystem of the islands. The Zero Fossil Fuels initiative for The Galapagos proposed by the Ecuadorian government as an alternative to reduce the use of fossil fuels in the islands, considers the replacement of diesel in thermoelectric generators, by Jatropha curcas vegetable oil. However, the Jatropha oil supply cannot entirely cover yet the demand for electricity generation in Galapagos. Within this context, the present work aims to provide an optimization model that can be used as a selection criterion for approving new Jatropha Curcas collection points in rural areas of Manabi-Ecuador. For this purpose, existing Jatropha collection points in Manabi were grouped under three regions: north (7 collection points), center (4 collection points) and south (9 collection points). Field work was carried out in every region in order to characterize the collection points, to establish local Jatropha supply and to determine transportation costs. Data collection was complemented using GIS software and an objective function was defined in order to determine the profit associated to Jatropha oil production. The market price of both Jatropha oil and residual cake, were considered for the total revenue; whereas Jatropha price, transportation and oil extraction costs were considered for the total cost. The tonnes of Jatropha fruit and seed, transported from collection points to the extraction plant, were considered as variables. The maximum and minimum amount of the collected Jatropha from each region constrained the optimization problem. The supply chain was optimized using linear programming in order to maximize the profits. Finally, a sensitivity analysis was performed in order to find a profit-based criterion for the acceptance of future collection points in Manabi. The maximum profit reached a value of $ 4,616.93 per year, which represented a total Jatropha collection of 62.3 tonnes Jatropha per year. The northern region of Manabi had the biggest collection share (69%), followed by the southern region (17%). The criteria for accepting new Jatropha collection points in the rural areas of Manabi can be defined by the current maximum profit of the zone and by the variation in the profit when collection points are removed one at a time. The definition of new feasible collection points plays a key role in the supply chain associated to Jatropha oil production. Therefore, a mathematical model that assists decision makers in establishing new collection points while assuring profitability, contributes to guarantee a continued Jatropha oil supply for Galapagos and a sustained economic growth in the rural areas of Ecuador.

Keywords: collection points, Jatropha curcas, linear programming, supply chain

Procedia PDF Downloads 400
219 Studies on Radio Frequency Sputtered Copper Zinc Tin Sulphide Absorber Layers for Thin Film Solar Cells

Authors: G. Balaji, R. Balasundaraprabhu, S. Prasanna, M. D. Kannan, K. Sivakumaran, David Mcilroy

Abstract:

Copper Zin tin sulphide (Cu2ZnSnS4 or CZTS) is found to be better alternative to Copper Indium gallium diselenide as absorber layers in thin film based solar cells due to the utilisation of earth-abundant materials in the midst of lower toxicity. In the present study, Cu2ZnSnS4 thin films were prepared on soda lime glass using (CuS, ZnS, SnS) targets and were deposited by three different stacking orders, using RF Magnetron sputtering. The substrate temperature was fixed at 300 °C during the depositions. CZTS thin films were characterized using X-ray diffraction, X-ray photoelectron spectroscopy, Raman spectroscopy and UV-Vis-NIR spectroscopy. All the samples exhibited X-ray peaks pertaining to (112) kesterite phase of CZTS, along with the presence of a predominant wurtzite CZTS phase. X-ray photoelectron spectroscopy revealed the presence of all the elements in all the samples. The change in stacking order clearly shows that it affects the structural and phase properties of the films. Relative atomic concentrations of Zn, Cu, Sn and S, which are determined by high-resolution XPS core level spectra integrated peak areas revealed that the CZTS films exhibit inhomogeneity in both stoichiometry and elemental composition. Raman spectroscopy studies on the film showed the presence of CZTS phase. The energy band gap of the CZTS thin films was found to be in the range of 1.5 eV to 1.6 eV. The films were then annealed at 450 °C for 5 hrs and it was found that the predominant nature of the X-ray peaks has transformed from Wurtzite to Kesterite phase which is highly desirable for absorber layers in thin film solar cells. The optimized CZTS layer was used as an absorber layer in thin film solar cells. ZnS and CdS were used as buffer layers which in turn prepared by Hot wall epitaxy technique. Gallium doped Zinc oxide was used as a transparent conducting oxide. The solar cell structure Glass/Mo/CZTS/CdS or ZnS/GZO has been fabricated, and solar cell parameters were measured.

Keywords: earth-abundant, Kesterite, RF sputtering, thin film solar cells

Procedia PDF Downloads 256
218 Thickness-Tunable Optical, Magnetic, and Dielectric Response of Lithium Ferrite Thin Film Synthesized by Pulsed Laser Deposition

Authors: Prajna Paramita Mohapatra, Pamu Dobbidi

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Lithium ferrite (LiFe5O8) has potential applications as a component of microwave magnetic devices such as circulators and monolithic integrated circuits. For efficient device applications, spinel ferrites in the form of thin films are highly required. It is necessary to improve their magnetic and dielectric behavior by optimizing the processing parameters during deposition. The lithium ferrite thin films are deposited on Pt/Si substrate using the pulsed laser deposition technique (PLD). As controlling the film thickness is the easiest parameter to tailor the strain, we deposited the thin films having different film thicknesses (160 nm, 200 nm, 240 nm) at oxygen partial pressure of 0.001 mbar. The formation of single phase with spinel structure (space group - P4132) is confirmed by the XRD pattern and the Rietveld analysis. The optical bandgap is decreased with the increase in thickness. FESEM confirmed the formation of uniform grains having well separated grain boundaries. Further, the film growth and the roughness are analyzed by AFM. The root-mean-square (RMS) surface roughness is decreased from 13.52 nm (160 nm) to 9.34 nm (240 nm). The room temperature magnetization is measured with a maximum field of 10 kOe. The saturation magnetization is enhanced monotonically with an increase in thickness. The magnetic resonance linewidth is obtained in the range of 450 – 780 Oe. The dielectric response is measured in the frequency range of 104 – 106 Hz and in the temperature range of 303 – 473 K. With an increase in frequency, the dielectric constant and the loss tangent of all the samples decreased continuously, which is a typical behavior of conventional dielectric material. The real part of the dielectric constant and the dielectric loss is increased with an increase in thickness. The contribution of grain and grain boundaries is also analyzed by employing the equivalent circuit model. The highest dielectric constant is obtained for the film having a thickness of 240 nm at 104 Hz. The obtained results demonstrate that desired response can be obtained by tailoring the film thickness for the microwave magnetic devices.

Keywords: PLD, optical response, thin films, magnetic response, dielectric response

Procedia PDF Downloads 68
217 Ultrasensitive Detection and Discrimination of Cancer-Related Single Nucleotide Polymorphisms Using Poly-Enzyme Polymer Bead Amplification

Authors: Lorico D. S. Lapitan Jr., Yihan Xu, Yuan Guo, Dejian Zhou

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The ability of ultrasensitive detection of specific genes and discrimination of single nucleotide polymorphisms is important for clinical diagnosis and biomedical research. Herein, we report the development of a new ultrasensitive approach for label-free DNA detection using magnetic nanoparticle (MNP) assisted rapid target capture/separation in combination with signal amplification using poly-enzyme tagged polymer nanobead. The sensor uses an MNP linked capture DNA and a biotin modified signal DNA to sandwich bind the target followed by ligation to provide high single-nucleotide polymorphism discrimination. Only the presence of a perfect match target DNA yields a covalent linkage between the capture and signal DNAs for subsequent conjugation of a neutravidin-modified horseradish peroxidase (HRP) enzyme through the strong biotin-nuetravidin interaction. This converts each captured DNA target into an HRP which can convert millions of copies of a non-fluorescent substrate (amplex red) to a highly fluorescent product (resorufin), for great signal amplification. The use of polymer nanobead each tagged with thousands of copies of HRPs as the signal amplifier greatly improves the signal amplification power, leading to greatly improved sensitivity. We show our biosensing approach can specifically detect an unlabeled DNA target down to 10 aM with a wide dynamic range of 5 orders of magnitude (from 0.001 fM to 100.0 fM). Furthermore, our approach has a high discrimination between a perfectly matched gene and its cancer-related single-base mismatch targets (SNPs): It can positively detect the perfect match DNA target even in the presence of 100 fold excess of co-existing SNPs. This sensing approach also works robustly in clinical relevant media (e.g. 10% human serum) and gives almost the same SNP discrimination ratio as that in clean buffers. Therefore, this ultrasensitive SNP biosensor appears to be well-suited for potential diagnostic applications of genetic diseases.

Keywords: DNA detection, polymer beads, signal amplification, single nucleotide polymorphisms

Procedia PDF Downloads 227
216 The Fabrication and Characterization of a Honeycomb Ceramic Electric Heater with a Conductive Coating

Authors: Siming Wang, Qing Ni, Yu Wu, Ruihai Xu, Hong Ye

Abstract:

Porous electric heaters, compared to conventional electric heaters, exhibit excellent heating performance due to their large specific surface area. Porous electric heaters employ porous metallic materials or conductive porous ceramics as the heating element. The former attains a low heating power with a fixed current due to the low electrical resistivity of metal. Although the latter can bypass the inherent challenges of porous metallic materials, the fabrication process of the conductive porous ceramics is complicated and high cost. This work proposed a porous ceramic electric heater with dielectric honeycomb ceramic as a substrate and surface conductive coating as a heating element. The conductive coating was prepared by the sol-gel method using silica sol and methyl trimethoxysilane as raw materials and graphite powder as conductive fillers. The conductive mechanism and degradation reason of the conductive coating was studied by electrical resistivity and thermal stability analysis. The heating performance of the proposed heater was experimentally investigated by heating air and deionized water. The results indicate that the electron transfer is achieved by forming the conductive network through the contact of the graphite flakes. With 30 wt% of graphite, the electrical resistivity of the conductive coating can be as low as 0.88 Ω∙cm. The conductive coating exhibits good electrical stability up to 500°C but degrades beyond 600°C due to the formation of many cracks in the coating caused by the weight loss and thermal expansion. The results also show that the working medium has a great influence on the volume power density of the heater. With air under natural convection as the working medium, the volume power density attains 640.85 kW/m3, which can be increased by 5 times when using deionized water as the working medium. The proposed honeycomb ceramic electric heater has the advantages of the simple fabrication method, low cost, and high volume power density, demonstrating great potential in the fluid heating field.

Keywords: conductive coating, honeycomb ceramic electric heater, high specific surface area, high volume power density

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215 Antioxidant Activity of Some Important Indigenous Plant Foods of the North Eastern Region of India

Authors: L. Bidyalakshmi, R. Ananthan, T. Longvah

Abstract:

Antioxidants are substances that can prevent or delay oxidative damage of lipids, proteins and nucleic acids by reactive oxygen species. These help in lowering incidence of degenerative diseases such as cancer, arthritis, atherosclerosis, heart disease, inflammation, brain dysfunction and acceleration of the ageing process. The north eastern part of India falls among the global hotspots of biodiversity. Over the years, the local communities in the region have developed ingenious uses of many wild plants within their environment as food sources. Many of these less familiar foods form an integral part of the diet of these communities, and some are traditionally valued for its therapeutic effects. So the study was carried to estimate the antioxidant activity of some of these indigenous foods. Twenty-eight indigenous plant foods were studied for their antioxidant activity. Antioxidant activities were determined by using DPPH (2, 2-diphenyl-1-picrylhydrazyl) assay, FRAP (Ferric Reducing Antioxidant Power) assay and SOSA (Super Oxide Scavenging Assay). Out of the twenty-eight plant foods, there were thirteen leafy vegetables, four fruits, five roots and tubers, four spices and two mushrooms. Water extract and methanol extract of the samples were used for the analysis. The leafy vegetable samples exhibited antioxidant capacity with IC50 ranging from 8-1414 mg/ml for lipid extract and 34-37878 mg/ml for aqueous extract in DPPH assay. Total FRAP value ranging from 58-1005 mmol FeSO4 Eq/100g of the sample, which is comparatively higher than the antioxidant capacity of some commonly consumed leafy vegetables. In SOSA, water extract of leafy vegetables show a range of 0.05-193.68 µmol ascorbic acid equivalent/g of the samples. While the methanol extract of the samples show 0.20-21.94 µmol Trolox equivalent/g of the samples. Polygonum barbatum, Wendlandia glabrata and Polygonum posumbu have higher antioxidant activity among the leafy vegetables analysed. Among the fruits, Rhus hookerii showed the highest antioxidant activities in both FRAP and SOSA methods while Spondias magnifera exhibited higher antioxidant activity in DPPH method. Alocasia cucullata exhibited higher antioxidant activity in DPPH and FRAP assays while Alpinia galanga showed higher antioxidant activity in SOSA assay when compared to the other samples of roots and tubers. Elsholtzia communis showed high antioxidant activity in all the three parameters among the spices. For the mushrooms, Pleurotus ostreatus exhibited higher antioxidant activity than Auricularia delicate in DPPH and SOSA. The samples analysed exhibited antioxidant activity at varying levels and some exhibited higher antioxidant activity than the commonly consumed foods. So consumption of these less familiar foods may play a role in preventing human disease in which free radicals are involved. Further studies on these food samples on phytonutrients and its contribution to the antioxidant activities are required.

Keywords: antioxidant activity, DPPH, FRAP, SOSA

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