Search results for: phosphate deposits
Commenced in January 2007
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Edition: International
Paper Count: 815

Search results for: phosphate deposits

125 Lacustrine Sediments of the Poljanska Locality in the Miocene Climatic Optimum North Croatian Basin, Croatia

Authors: Marijan KovačIć, Davor Pavelić, Darko Tibljaš, Ivo Galić, Frane Marković, Ivica PavičIć

Abstract:

The North Croatian Basin (NCB) occupies the southwestern part of the Pannonian Basin System and belongs to the Central Paratethys realm. In a quarry near the village of Poljanska, on the southern slopes of Mt. Papuk in eastern Croatia, a 40-meter-thick section is exposed, consisting of well-bedded, mixed, carbonate-siliciclastic deposits with occurrences of pyroclastics. Sedimentological investigation indicates that a salina lake developed in the central NCB during the late early Miocene. Field studies and mineralogical and petrological analyses indicate that alternations of laminated crypto- characterize the lower part of the section to microcrystalline dolomite and analcimolite (sedimentary rocks composed essentially of authigenic analcime) associated with tuffites and marls. The pyroclastic material is a product of volcanic activity at the end of the early Miocene, while the formation of analcime, the zeolite group mineral, is a result of an alteration of pyroclastic material in an alkaline lacustrine environment. These sediments were deposited in a shallow, hydrologically closed lake that was controlled by an arid climate during the first phase of its development. The middle part of the section consists of dolomites interbedded with analcimolites and sandstones. The sandstone beds are a result of the increased supply of clastic material derived from the locally uplifted metamorphic and granitoid basement. The emplacement of sandstones and dolomites reflects a distinct alternation of hydrologically open and closed lacustrine environments controlled by the frequent alternation of humid and arid climates, representing the second phase of lake development. The siliciclastics of the third phase of lake development were deposited during the Middle Miocene in a hydrologically mostly open lake. All lacustrine deposition coincides with the Miocene Climatic Optimum, which was characterized by a hot and warm climate. The sedimentological data confirm the mostly wet conditions previously identified by paleobotanical studies in the region. The exception is the relatively long interval of arid climate in the late early Miocene that controlled the first phase of lake evolution, i.e., the salina-type lake.

Keywords: early Miocene, Pannonian basin System, pyroclastics, salina-type lake

Procedia PDF Downloads 195
124 Algae Biofertilizers Promote Sustainable Food Production and Nutrient Efficiency: An Integrated Empirical-Modeling Study

Authors: Zeenat Rupawalla, Nicole Robinson, Susanne Schmidt, Sijie Li, Selina Carruthers, Elodie Buisset, John Roles, Ben Hankamer, Juliane Wolf

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Agriculture has radically changed the global biogeochemical cycle of nitrogen (N). Fossil fuel-enabled synthetic N-fertiliser is a foundation of modern agriculture but applied to soil crops only use about half of it. To address N-pollution from cropping and the large carbon and energy footprint of N-fertiliser synthesis, new technologies delivering enhanced energy efficiency, decarbonisation, and a circular nutrient economy are needed. We characterised algae fertiliser (AF) as an alternative to synthetic N-fertiliser (SF) using empirical and modelling approaches. We cultivated microalgae in nutrient solution and modelled up-scaled production in nutrient-rich wastewater. Over four weeks, AF released 63.5% of N as ammonium and nitrate, and 25% of phosphorous (P) as phosphate to the growth substrate, while SF released 100% N and 20% P. To maximise crop N-use and minimise N-leaching, we explored AF and SF dose-response-curves with spinach in glasshouse conditions. AF-grown spinach produced 36% less biomass than SF-grown plants due to AF’s slower and linear N-release, while SF resulted in 5-times higher N-leaching loss than AF. Optimised blends of AF and SF boosted crop yield and minimised N-loss due to greater synchrony of N-release and crop uptake. Additional benefits of AF included greener leaves, lower leaf nitrate concentration, and higher microbial diversity and water holding capacity in the growth substrate. Life-cycle-analysis showed that replacing the most effective SF dosage with AF lowered the carbon footprint of fertiliser production from 2.02 g CO₂ (C-producing) to -4.62 g CO₂ (C-sequestering), with a further 12% reduction when AF is produced on wastewater. Embodied energy was lowest for AF-SF blends and could be reduced by 32% when cultivating algae on wastewater. We conclude that (i) microalgae offer a sustainable alternative to synthetic N-fertiliser in spinach production and potentially other crop systems, and (ii) microalgae biofertilisers support the circular nutrient economy and several sustainable development goals.

Keywords: bioeconomy, decarbonisation, energy footprint, microalgae

Procedia PDF Downloads 120
123 Bed Evolution under One-Episode Flushing in a Truck Sewer in Paris, France

Authors: Gashin Shahsavari, Gilles Arnaud-Fassetta, Alberto Campisano, Roberto Bertilotti, Fabien Riou

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Sewer deposits have been identified as a major cause of dysfunctions in combined sewer systems regarding sewer management, which induces different negative consequents resulting in poor hydraulic conveyance, environmental damages as well as worker’s health. In order to overcome the problematics of sedimentation, flushing has been considered as the most operative and cost-effective way to minimize the sediments impacts and prevent such challenges. Flushing, by prompting turbulent wave effects, can modify the bed form depending on the hydraulic properties and geometrical characteristics of the conduit. So far, the dynamics of the bed-load during high-flow events in combined sewer systems as a complex environment is not well understood, mostly due to lack of measuring devices capable to work in the “hostile” in combined sewer system correctly. In this regards, a one-episode flushing issue from an opening gate valve with weir function was carried out in a trunk sewer in Paris to understanding its cleansing efficiency on the sediments (thickness: 0-30 cm). During more than 1h of flushing within 5 m distance in downstream of this flushing device, a maximum flowrate and a maximum level of water have been recorded at 5 m in downstream of the gate as 4.1 m3/s and 2.1 m respectively. This paper is aimed to evaluate the efficiency of this type of gate for around 1.1 km (from the point -50 m to +1050 m in downstream from the gate) by (i) determining bed grain-size distribution and sediments evolution through the sewer channel, as well as their organic matter content, and (ii) identifying sections that exhibit more changes in their texture after the flush. For the first one, two series of sampling were taken from the sewer length and then analyzed in laboratory, one before flushing and second after, at same points among the sewer channel. Hence, a non-intrusive sampling instrument has undertaken to extract the sediments smaller than the fine gravels. The comparison between sediments texture after the flush operation and the initial state, revealed the most modified zones by the flush effect, regarding the sewer invert slope and hydraulic parameters in the zone up to 400 m from the gate. At this distance, despite the increase of sediment grain-size rages, D50 (median grain-size) varies between 0.6 mm and 1.1 mm compared to 0.8 mm and 10 mm before and after flushing, respectively. Overall, regarding the sewer channel invert slope, results indicate that grains smaller than sands (< 2 mm) are more transported to downstream along about 400 m from the gate: in average 69% before against 38% after the flush with more dispersion of grain-sizes distributions. Furthermore, high effect of the channel bed irregularities on the bed material evolution has been observed after the flush.

Keywords: bed-load evolution, combined sewer systems, flushing efficiency, sediments transport

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122 Calcein Release from Liposomes Mediated by Phospholipase A₂ Activity: Effect of Cholesterol and Amphipathic Di and Tri Blocks Copolymers

Authors: Marco Soto-Arriaza, Eduardo Cena-Ahumada, Jaime Melendez-Rojel

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Background: Liposomes have been widely used as a model of lipid bilayer to study the physicochemical properties of biological membrane, encapsulation, transport and release of different molecules. Furthermore, extensive research has focused on improving the efficiency in the transport of drugs, developing tools that improve the release of the encapsulated drug from liposomes. In this context, the enzymatic activity of PLA₂, despite having been shown to be an effective tool to promote the release of drugs from liposomes, is still an open field of research. Aim: The aim of the present study is to explore the effect of cholesterol (Cho) and amphipathic di- and tri-block copolymers, on calcein release mediated by enzymatic activity of PLA2 in Dipalmitoylphosphatidylcholine (DPPC) liposomes under physiological conditions. Methods: Different dispersions of DPPC, cholesterol, di-block POE₄₅-PCL₅₂ or tri-block PCL₁₂-POE₄₅-PCL₁₂ were prepared by the extrusion method after five freezing/thawing cycles; in Phosphate buffer 10mM pH 7.4 in presence of calcein. DPPC liposomes/Calcein were centrifuged at 15000rpm 10 min to separate free calcein. Enzymatic activity assays of PLA₂ were performed at 37°C using the TBS buffer pH 7.4. The size distribution, polydispersity, Z-potential and Calcein encapsulation of DPPC liposomes was monitored. Results: PLA₂ activity showed a slower kinetic of calcein release up to 20 mol% of cholesterol, evidencing a minimum at 10 mol% and then a maximum at 18 mol%. Regardless of the percentage of cholesterol, up to 18 mol% a one-hundred percentage release of calcein was observed. At higher cholesterol concentrations, PLA₂ showed to be inefficient or not to be involved in calcein release. In assays where copolymers were added in a concentration lower than their cmc, a similar behavior to those showed in the presence of Cho was observed, that is a slower kinetic in calcein release. In both experimental approaches, a one-hundred percentage of calcein release was observed. PLA₂ was shown to be sensitive to the 4-(4-Octadecylphenyl)-4-oxobutenoic acid inhibitor and calcium, reducing the release of calcein to 0%. Cell viability of HeLa cells decreased 7% in the presence of DPPC liposomes after 3 hours of incubation and 17% and 23% at 5 and 15 hours, respectively. Conclusion: Calcein release from DPPC liposomes, mediated by PLA₂ activity, depends on the percentage of cholesterol and the presence of copolymers. Both, cholesterol up to 20 mol% and copolymers below it cmc could be applied to the regulation of the kinetics of antitumoral drugs release without inducing cell toxicity per se.

Keywords: amphipathic copolymers, calcein release, cholesterol, DPPC liposome, phospholipase A₂

Procedia PDF Downloads 139
121 In vitro Evaluation of Capsaicin Patches for Transdermal Drug Delivery

Authors: Alija Uzunovic, Sasa Pilipovic, Aida Sapcanin, Zahida Ademovic, Berina Pilipović

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Capsaicin is a naturally occurring alkaloid extracted from capsicum fruit extracts of different of Capsicum species. It has been employed topically to treat many diseases such as rheumatoid arthritis, osteoarthritis, cancer pain and nerve pain in diabetes. The high degree of pre-systemic metabolism of intragastrical capsaicin and the short half-life of capsaicin by intravenous administration made topical application of capsaicin advantageous. In this study, we have evaluated differences in the dissolution characteristics of capsaicin patch 11 mg (purchased from market) at different dissolution rotation speed. The proposed patch area is 308 cm2 (22 cm x 14 cm; it contains 36 µg of capsaicin per square centimeter of adhesive). USP Apparatus 5 (Paddle Over Disc) is used for transdermal patch testing. The dissolution study was conducted using USP apparatus 5 (n=6), ERWEKA DT800 dissolution tester (paddle-type) with addition of a disc. The fabricated patch of 308 cm2 is to be cut into 9 cm2 was placed against a disc (delivery side up) retained with the stainless-steel screen and exposed to 500 mL of phosphate buffer solution pH 7.4. All dissolution studies were carried out at 32 ± 0.5 °C and different rotation speed (50± 5; 100± 5 and 150± 5 rpm). 5 ml aliquots of samples were withdrawn at various time intervals (1, 4, 8 and 12 hours) and replaced with 5 ml of dissolution medium. Withdrawn were appropriately diluted and analyzed by reversed-phase liquid chromatography (RP-LC). A Reversed Phase Liquid Chromatography (RP-LC) method has been developed, optimized and validated for the separation and quantitation of capsaicin in a transdermal patch. The method uses a ProntoSIL 120-3-C18 AQ 125 x 4,0 mm (3 μm) column maintained at 600C. The mobile phase consisted of acetonitrile: water (50:50 v/v), the flow rate of 0.9 mL/min, the injection volume 10 μL and the detection wavelength 222 nm. The used RP-LC method is simple, sensitive and accurate and can be applied for fast (total chromatographic run time was 4.0 minutes) and simultaneous analysis of capsaicin and dihydrocapsaicin in a transdermal patch. According to the results obtained in this study, we can conclude that the relative difference of dissolution rate of capsaicin after 12 hours was elevated by increase of dissolution rotation speed (100 rpm vs 50 rpm: 84.9± 11.3% and 150 rpm vs 100 rpm: 39.8± 8.3%). Although several apparatus and procedures (USP apparatus 5, 6, 7 and a paddle over extraction cell method) have been used to study in vitro release characteristics of transdermal patches, USP Apparatus 5 (Paddle Over Disc) could be considered as a discriminatory test. would be able to point out the differences in the dissolution rate of capsaicin at different rotation speed.

Keywords: capsaicin, in vitro, patch, RP-LC, transdermal

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120 Using Hemicellulosic Liquor from Sugarcane Bagasse to Produce Second Generation Lactic Acid

Authors: Regiane A. Oliveira, Carlos E. Vaz Rossell, Rubens Maciel Filho

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Lactic acid, besides a valuable chemical may be considered a platform for other chemicals. In fact, the feasibility of hemicellulosic sugars as feedstock for lactic acid production process, may represent the drop of some of the barriers for the second generation bioproducts, especially bearing in mind the 5-carbon sugars from the pre-treatment of sugarcane bagasse. Bearing this in mind, the purpose of this study was to use the hemicellulosic liquor from sugarcane bagasse as a substrate to produce lactic acid by fermentation. To release of sugars from hemicellulose it was made a pre-treatment with a diluted sulfuric acid in order to obtain a xylose's rich liquor with low concentration of inhibiting compounds for fermentation (≈ 67% of xylose, ≈ 21% of glucose, ≈ 10% of cellobiose and arabinose, and around 1% of inhibiting compounds as furfural, hydroxymethilfurfural and acetic acid). The hemicellulosic sugars associated with 20 g/L of yeast extract were used in a fermentation process with Lactobacillus plantarum to produce lactic acid. The fermentation process pH was controlled with automatic injection of Ca(OH)2 to keep pH at 6.00. The lactic acid concentration remained stable from the time when the glucose was depleted (48 hours of fermentation), with no further production. While lactic acid is produced occurs the concomitant consumption of xylose and glucose. The yield of fermentation was 0.933 g lactic acid /g sugars. Besides, it was not detected the presence of by-products, what allows considering that the microorganism uses a homolactic fermentation to produce its own energy using pentose-phosphate pathway. Through facultative heterofermentative metabolism the bacteria consume pentose, as is the case of L. plantarum, but the energy efficiency for the cell is lower than during the hexose consumption. This implies both in a slower cell growth, as in a reduction in lactic acid productivity compared with the use of hexose. Also, L. plantarum had shown to have a capacity for lactic acid production from hemicellulosic hydrolysate without detoxification, which is very attractive in terms of robustness for an industrial process. Xylose from hydrolyzed bagasse and without detoxification is consumed, although the hydrolyzed bagasse inhibitors (especially aromatic inhibitors) affect productivity and yield of lactic acid. The use of sugars and the lack of need for detoxification of the C5 liquor from sugarcane bagasse hydrolyzed is a crucial factor for the economic viability of second generation processes. Taking this information into account, the production of second generation lactic acid using sugars from hemicellulose appears to be a good alternative to the complete utilization of sugarcane plant, directing molasses and cellulosic carbohydrates to produce 2G-ethanol, and hemicellulosic carbohydrates to produce 2G-lactic acid.

Keywords: fermentation, lactic acid, hemicellulosic sugars, sugarcane

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119 Selection of Suitable Reference Genes for Assessing Endurance Related Traits in a Native Pony Breed of Zanskar at High Altitude

Authors: Prince Vivek, Vijay K. Bharti, Manishi Mukesh, Ankita Sharma, Om Prakash Chaurasia, Bhuvnesh Kumar

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High performance of endurance in equid requires adaptive changes involving physio-biochemical, and molecular responses in an attempt to regain homeostasis. We hypothesized that the identification of the suitable reference genes might be considered for assessing of endurance related traits in pony at high altitude and may ensure for individuals struggling to potent endurance trait in ponies at high altitude. A total of 12 mares of ponies, Zanskar breed, were divided into three groups, group-A (without load), group-B, (60 Kg) and group-C (80 Kg) on backpack loads were subjected to a load carry protocol, on a steep climb of 4 km uphill, and of gravel, uneven rocky surface track at an altitude of 3292 m to 3500 m (endpoint). Blood was collected before and immediately after the load carry on sodium heparin anticoagulant, and the peripheral blood mononuclear cell was separated for total RNA isolation and thereafter cDNA synthesis. Real time-PCR reactions were carried out to evaluate the mRNAs expression profile of a panel of putative internal control genes (ICGs), related to different functional classes, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β₂ microglobulin (β₂M), β-actin (ACTB), ribosomal protein 18 (RS18), hypoxanthine-guanine phosophoribosyltransferase (HPRT), ubiquitin B (UBB), ribosomal protein L32 (RPL32), transferrin receptor protein (TFRC), succinate dehydrogenase complex subunit A (SDHA) for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of native pony’s. Three different algorithms, geNorm, NormFinder, and BestKeeper software, were used to evaluate the stability of reference genes. The result showed that GAPDH was best stable gene and stability value for the best combination of two genes was observed TFRC and β₂M. In conclusion, the geometric mean of GAPDH, TFRC and β₂M might be used for accurate normalization of transcriptional data for assessing endurance related traits in Zanskar ponies during load carrying.

Keywords: endurance exercise, ubiquitin B (UBB), β₂ microglobulin (β₂M), high altitude, Zanskar ponies, reference gene

Procedia PDF Downloads 119
118 Experimental Design in Extraction of Pseudomonas sp. Protease from Fermented Broth by Polyethylene Glycol/Citrate Aqueous Two-Phase System

Authors: Omar Pillaca-Pullo, Arturo Alejandro-Paredes, Carol Flores-Fernandez, Marijuly Sayuri Kina, Amparo Iris Zavaleta

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Aqueous two-phase system (ATPS) is an interesting alternative for separating industrial enzymes due to it is easy to scale-up and low cost. Polyethylene glycol (PEG) mixed with potassium phosphate or magnesium sulfate is one of the most frequently polymer/salt ATPS used, but the consequences of its use is a high concentration of phosphates and sulfates in wastewater causing environmental issues. Citrate could replace these inorganic salts due to it is biodegradable and does not produce toxic compounds. On the other hand, statistical design of experiments is widely used for ATPS optimization and it allows to study the effects of the involved variables in the purification, and to estimate their significant effects on selected responses and interactions. The 24 factorial design with four central points (20 experiments) was employed to study the partition and purification of proteases produced by Pseudomonas sp. in PEG/citrate ATPS system. ATPS was prepared with different sodium citrate concentrations [14, 16 and 18% (w/w)], pH values (7, 8 and 9), PEG molecular weight (2,000; 4,000 and 6,000 g/mol) and PEG concentrations [18, 20 and 22 % (w/w)]. All system components were mixed with 15% (w/w) of the fermented broth and deionized water was added to a final weight of 12.5 g. Then, the systems were mixed and kept at room temperature until to reach two-phases separation. Volumes of the top and bottom phases were measured, and aliquots from both phases were collected for subsequent proteolytic activity and total protein determination. Influence of variables such as PEG molar mass (MPEG), PEG concentration (CPEG), citrate concentration (CSal) and pH were evaluated on the following responses: purification factor (PF), activity yield (Y), partition coefficient (K) and selectivity (S). STATISTICA program version 10 was used for the analysis. According to the obtained results, higher levels of CPEG and MPEG had a positive effect on extraction, while pH did not influence on the process. On the other hand, the CSal could be related with low values of Y because of the citrate ions have a negative effect on solubility and enzymatic structure. The optimum values of Y (66.4 %), PF (1.8), K (5.5) and S (4.3) were obtained at CSal (18%), MPEG (6,000 g/mol), CPEG (22%) and pH 9. These results indicated that the PEG/citrate system is accurate to purify these Pseudomonas sp. proteases from fermented broth as a first purification step.

Keywords: citrate, polyethylene glycol, protease, Pseudomonas sp

Procedia PDF Downloads 180
117 Freshwater Source of Sapropel for Healthcare

Authors: Ilona Pavlovska, Aneka Klavina, Agris Auce, Ivars Vanadzins, Alise Silova, Laura Komarovska, Linda Paegle, Baiba Silamikele, Linda Dobkevica

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Freshwater sapropel is a common material formed by complex biological transformations of Holocene sediments in the water basement of the lakes in Latvia that has the potential to be used as medical mud. Sapropel forms over a long period in shallow waters by slowly decomposing organic sediment and has different compositions depending on the location of the source, surroundings, the water regime, etc. Official geological survey of Latvia lakes, from Latvian lake database (ezeri.lv), used in the selection of the area of the exploration. The multifunctional effect of sapropel on the whole organism explained by its complex chemical and biological structure. This unique, organic substance and its ability to maintain heat for a long time ensures deep tissue warming and has a positive effect on the treatment of various joint and skin diseases. Sapropel is a valuable resource with multiple areas of application. Investigation of sapropel sediments and survey of the five sites selected according to the criteria performed in the current study. Also, our study includes sampling at different depths and their initial treatment, evaluation of external signs, and study of physical-chemical parameters, as well as analysis of biochemical parameters and evaluation of microbiological indicators. The main selection criteria were sapropel deposits depth, hydrological regime, the history of agriculture next to the lake, and the potential exposure to industrial waste. One hundred and five sapropel samples obtained from five lakes (Audzelu, Dunakla, Ivusku, Zielu, and Mazars Kivdalova) during the wintertime. The main goal of the study is to carry out detailed and systematic research on the medical properties of sapropel to be obtained in Latvia, to promote its scientifically based use in balneology, to develop new medical procedures and services, and to promote the development of new exportable products. Latvian freshwater sapropel could be used as raw material for getting sapropel extract and use it as a remedy. All mentioned above brings us to the main question for sapropel usage in medicine, balneology, and pharmacy “how to develop quality criteria for raw sapropel and its extracts. The research was co-financed by the project "Analysis of characteristics of medical sapropel and its usage for medical purposes and elaboration of industrial extraction methods" No.1.1.1.1/16/A/165.

Keywords: balneology, extracts, freshwater sapropel, Latvian lakes, medical mud, sapropel

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116 Statistical Design of Central Point for Evaluate the Combination of PH and Cinnamon Essential Oil on the Antioxidant Activity Using the ABTS Technique

Authors: H. Minor-Pérez, A. M. Mota-Silva, S. Ortiz-Barrios

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Substances of vegetable origin with antioxidant capacity have a high potential for application on the conservation of some foods, can prevent or reduce for example oxidation of lipids. However a food is a complex system whose wide variety of components wich can reduce or eliminate this antioxidant capacity. The antioxidant activity can be determined with the ABTS technique. The radical ABTS+ is generated from the acid 2, 2´ - Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). This radical is a composite color bluish-green, stable and with a spectrum of absorption into the UV-visible. The addition of antioxidants causes discoloration, value that can be reported as a percentage of inhibition of the cation radical ABTS+. The objective of this study was evaluated the effect of the combination of the pH and the essential oil of cinnamon (EOC) on inhibition of the radical ABTS+, using statistical design of central point (Design Expert) to obtain mathematical models that describe this phenomenon. Were evaluated 17 treatments with combinations of pH 5, 6 and 7 (citrate-phosphate buffer) and the concentration of essential oil of cinnamon (C): 0 µg/mL, 100 µg/mL and 200 µg/mL. The samples were analyzed using the ABTS technique. The reagent was dissolved in methanol 80% to standardized the absorbance to 0.7 +/- 0.1 at 754 nm. Then samples were mixed with reagent standardized ABTS and after 1 min and 7 min absorbance was read for each treatment at 754 nm. Was used a curve pattern with vitamin C and reported the values as inhibition (%) of radical ABTS+. The statistical analysis shows the experimental results were adjusted to a quadratic model, to the times of 1 min and 7 min. This model describes the influence of the factors investigated independently: pH and cinnamon essential oil (µg/mL) and the effect of the interaction between pH*C, as well as the square of the pH2 and C2. The model obtained was Y = 10.33684 - 3.98118*pH + 1.17031*C + 0.62745*pH2 - 3.26675*10-3*C2 - 0.013112*pH*C, where Y is the response variable. The coefficient of determination was 0.9949 for 1 min. The equation was obtained at 7 min and = - 10.89710 + 1.52341*pH + 1.32892*C + 0.47953*pH2 - 3.56605*10- *C2 - 0.034687*pH*C. The coefficient of determination was 0.9970. This means that only 1% of the total variation is not explained by the developed models. At 100 µg/mL of EOC was obtained an inhibition percentage of 80%, 84% and 97% for the pH values of 5,6 and 7 respectively, while a value of 200 µg/mL the inhibition (%) was very similar for the treatments. In these values of pH was obtained an inhibition close 97%. In conclusion the pH does not have a significant effect on the antioxidant capacity, while the concentration of EOC was decisive for the antioxidant capacity. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: antioxidant activity, ABTS technique, essential oil of cinnamon, mathematical models

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115 Intrathecal: Not Intravenous Administration of Evans Blue Reduces Pain Behavior in Neuropathic Rats

Authors: Kun Hua O., Dong Woon Kim, Won Hyung Lee

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Introduction: Neuropathic pain induced by spinal or peripheral nerve injury is highly resistant to common painkillers, nerve blocks, and other pain management approaches. Recently, several new therapeutic drug candidates have been developed to control neuropathic pain. In this study, we used the spinal nerve L5 ligation (SNL) model to investigate the ability of intrathecal or intravenous Evans blue to decrease pain behavior and to study the relationship between Evans blue and the neural structure of pain transmission. Method: Neuropathic pain (allodynia) of the left hind paw was induced by unilateral SNL in Sprague-Dawley rats(n=10) in each group. Evans blue (5, 15, 50μg/10μl) or phosphate buffer saline(PBS,10μl) was injected intrathecally at 3days post-ligation or intravenously(1mg/200 μl) 3days and 5days post-ligation . Mechanical sensitivity was assessed using Von Frey filaments at 3 days post-ligation and at 2 hours, days 1, 2, 3, 5,7 after intrathecal Evans blue injection, and on days 2, 4, 7, and 11 at 14 days after intravenous injection. In the intrathecal group, microglia and glutaminergic neurons in the dorsal horn and VNUT(vesicular nucleotide transporter) in the dorsal root ganglia were tested to evaluate co-staining with Evans blue. The experimental procedures were performed in accordance with the animal care guideline of the Korean Academy of Medical Science(Animal ethic committee of Chungnam National University Hospital: CNUH-014-A0005-1). Results: Tight ligation of the L5 spinal nerve induced allodynia in the left hind paw 3 days post-ligation. Intrathecal Evans blue most significantly(P<0.001) alleviated allodynia at 2 days after intrathecal, but not an intravenous injection. Glutaminergic neurons in the dorsal horn and VNUT in the dorsal root ganglia were co-stained with Evans blue. On the other hand, microglia in the dorsal horn were partially co-stained with Evans blue. Conclusion: We confirmed that Evans blue might have an analgesic effect through the central nervous system, not another system in neuropathic pain of the SNL animal model. These results suggest Evans blue may be a potential new drug for the treatment of chronic pain. This research was supported by the National Research Foundation of Korea (NRF-2020R1A2C100757512), funded by the Ministry of Education.

Keywords: neuropathic pain, Evas blue, intrathecal, intravenous

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114 Computational Approach to Identify Novel Chemotherapeutic Agents against Multiple Sclerosis

Authors: Syed Asif Hassan, Tabrej Khan

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Multiple sclerosis (MS) is a chronic demyelinating autoimmune disorder, of the central nervous system (CNS). In the present scenario, the current therapies either do not halt the progression of the disease or have side effects which limit the usage of current Disease Modifying Therapies (DMTs) for a longer period of time. Therefore, keeping the current treatment failure schema, we are focusing on screening novel analogues of the available DMTs that specifically bind and inhibit the Sphingosine1-phosphate receptor1 (S1PR1) thereby hindering the lymphocyte propagation toward CNS. The novel drug-like analogs molecule will decrease the frequency of relapses (recurrence of the symptoms associated with MS) with higher efficacy and lower toxicity to human system. In this study, an integrated approach involving ligand-based virtual screening protocol (Ultrafast Shape Recognition with CREDO Atom Types (USRCAT)) to identify the non-toxic drug like analogs of the approved DMTs were employed. The potency of the drug-like analog molecules to cross the Blood Brain Barrier (BBB) was estimated. Besides, molecular docking and simulation using Auto Dock Vina 1.1.2 and GOLD 3.01 were performed using the X-ray crystal structure of Mtb LprG protein to calculate the affinity and specificity of the analogs with the given LprG protein. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has a higher hypothetical affinity, also has greater negative value. Further, the non-specific ligands were screened out using the structural filter proposed by Baell and Holloway. Based on the USRCAT, Lipinski’s values, toxicity and BBB analysis, the drug-like analogs of fingolimod and BG-12 showed that RTL and CHEMBL1771640, respectively are non-toxic and permeable to BBB. The successful docking and DSX analysis showed that RTL and CHEMBL1771640 could bind to the binding pocket of S1PR1 receptor protein of human with greater affinity than as compared to their parent compound (Fingolimod). In this study, we also found that all the drug-like analogs of the standard MS drugs passed the Bell and Holloway filter.

Keywords: antagonist, binding affinity, chemotherapeutics, drug-like, multiple sclerosis, S1PR1 receptor protein

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113 Thorium Resources of Georgia – Is It Its Future Energy ?

Authors: Avtandil Okrostsvaridze, Salome Gogoladze

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In the light of exhaustion of hydrocarbon reserves of new energy resources, its search is of vital importance problem for the modern civilization. At the time of energy resource crisis, the radioactive element thorium (232Th) is considered as the main energy resource for the future of our civilization. Modern industry uses thorium in high-temperature and high-tech tools, but the most important property of thorium is that like uranium it can be used as fuel in nuclear reactors. However, thorium has a number of advantages compared to this element: Its concentration in the earth crust is 4-5 times higher than uranium; extraction and enrichment of thorium is much cheaper than of uranium; it is less radioactive; its waste products complete destruction is possible; thorium yields much more energy than uranium. Nowadays, developed countries, among them India and China, have started intensive work for creation of thorium nuclear reactors and intensive search for thorium reserves. It is not excluded that in the next 10 years these reactors will completely replace uranium reactors. Thorium ore mineralization is genetically related to alkaline-acidic magmatism. Thorium accumulations occur as in endogen marked as in exogenous conditions. Unfortunately, little is known about the reserves of this element in Georgia, as planned prospecting-exploration works of thorium have never been carried out here. Although, 3 ore occurrences of this element are detected: 1) In the Greater Caucasus Kakheti segment, in the hydrothermally altered rocks of the Lower Jurassic clay-shales, where thorium concentrations varied between 51 - 3882g/t; 2) In the eastern periphery of the Dzirula massif, in the hydrothermally alteration rocks of the cambrian quartz-diorite gneisses, where thorium concentrations varied between 117-266 g/t; 3) In active contact zone of the Eocene volcanites and syenitic intrusive in Vakijvari ore field of the Guria region, where thorium concentrations varied between 185 – 428 g/t. In addition, geological settings of the areas, where thorium occurrences were fixed, give a theoretical basis on possible accumulation of practical importance thorium ores. Besides, the Black Sea Guria region magnetite sand which is transported from Vakijvari ore field, should contain significant reserves of thorium. As the research shows, monazite (thorium containing mineral) is involved in magnetite in the form of the thinnest inclusions. The world class thorium deposit concentrations of this element vary within the limits of 50-200 g/t. Accordingly, on the basis of these data, thorium resources found in Georgia should be considered as perspective ore deposits. Generally, we consider that complex investigation of thorium should be included into the sphere of strategic interests of the state, because future energy of Georgia, will probably be thorium.

Keywords: future energy, Georgia, ore field, thorium

Procedia PDF Downloads 478
112 Optimization of Mechanical Properties of Alginate Hydrogel for 3D Bio-Printing Self-Standing Scaffold Architecture for Tissue Engineering Applications

Authors: Ibtisam A. Abbas Al-Darkazly

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In this study, the mechanical properties of alginate hydrogel material for self-standing 3D scaffold architecture with proper shape fidelity are investigated. In-lab built 3D bio-printer extrusion-based technology is utilized to fabricate 3D alginate scaffold constructs. The pressure, needle speed and stage speed are varied using a computer-controlled system. The experimental result indicates that the concentration of alginate solution, calcium chloride (CaCl2) cross-linking concentration and cross-linking ratios lead to the formation of alginate hydrogel with various gelation states. Besides, the gelling conditions, such as cross-linking reaction time and temperature also have a significant effect on the mechanical properties of alginate hydrogel. Various experimental tests such as the material gelation, the material spreading and the printability test for filament collapse as well as the swelling test were conducted to evaluate the fabricated 3D scaffold constructs. The result indicates that the fabricated 3D scaffold from composition of 3.5% wt alginate solution, that is prepared in DI water and 1% wt CaCl2 solution with cross-linking ratios of 7:3 show good printability and sustain good shape fidelity for more than 20 days, compared to alginate hydrogel that is prepared in a phosphate buffered saline (PBS). The fabricated self-standing 3D scaffold constructs measured 30 mm × 30 mm and consisted of 4 layers (n = 4) show good pore geometry and clear grid structure after printing. In addition, the percentage change of swelling degree exhibits high swelling capability with respect to time. The swelling test shows that the geometry of 3D alginate-scaffold construct and of the macro-pore are rarely changed, which indicates the capability of holding the shape fidelity during the incubation period. This study demonstrated that the mechanical and physical properties of alginate hydrogel could be tuned for a 3D bio-printing extrusion-based system to fabricate self-standing 3D scaffold soft structures. This 3D bioengineered scaffold provides a natural microenvironment present in the extracellular matrix of the tissue, which could be seeded with the biological cells to generate the desired 3D live tissue model for in vitro and in vivo tissue engineering applications.

Keywords: biomaterial, calcium chloride, 3D bio-printing, extrusion, scaffold, sodium alginate, tissue engineering

Procedia PDF Downloads 95
111 Biosorption of Nickel by Penicillium simplicissimum SAU203 Isolated from Indian Metalliferous Mining Overburden

Authors: Suchhanda Ghosh, A. K. Paul

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Nickel, an industrially important metal is not mined in India, due to the lack of its primary mining resources. But, the chromite deposits occurring in the Sukinda and Baula-Nuasahi region of Odhisa, India, is reported to contain around 0.99% of nickel entrapped in the goethite matrix of the lateritic iron rich ore. Weathering of the dumped chromite mining overburden often leads to the contamination of the ground as well as the surface water with toxic nickel. Microbes inherent to this metal contaminated environment are reported to be capable of removal as well as detoxification of various metals including nickel. Nickel resistant fungal isolates obtained in pure form from the metal rich overburden were evaluated for their potential to biosorb nickel by using their dried biomass. Penicillium simplicissimum SAU203 was the best nickel biosorbant among the 20 fungi tested and was capable to sorbing 16.85 mg Ni/g biomass from a solution containing 50 mg/l of Ni. The identity of the isolate was confirmed using 18S rRNA gene analysis. The sorption capacity of the isolate was further standardized following Langmuir and Freundlich adsorption isotherm models and the results reflected energy efficient sorption. Fourier-transform infrared spectroscopy studies of the nickel loaded and control biomass in a comparative basis revealed the involvement of hydroxyl, amine and carboxylic groups in Ni binding. The sorption process was also optimized for several standard parameters like initial metal ion concentration, initial sorbet concentration, incubation temperature and pH, presence of additional cations and pre-treatment of the biomass by different chemicals. Optimisation leads to significant improvements in the process of nickel biosorption on to the fungal biomass. P. simplicissimum SAU203 could sorb 54.73 mg Ni/g biomass with an initial Ni concentration of 200 mg/l in solution and 21.8 mg Ni/g biomass with an initial biomass concentration of 1g/l solution. Optimum temperature and pH for biosorption was recorded to be 30°C and pH 6.5 respectively. Presence of Zn and Fe ions improved the sorption of Ni(II), whereas, cobalt had a negative impact. Pre-treatment of biomass with various chemical and physical agents has affected the proficiency of Ni sorption by P. simplicissimum SAU203 biomass, autoclaving as well as treatment of biomass with 0.5 M sulfuric acid and acetic acid reduced the sorption as compared to the untreated biomass, whereas, NaOH and Na₂CO₃ and Twin 80 (0.5 M) treated biomass resulted in augmented metal sorption. Hence, on the basis of the present study, it can be concluded that P. simplicissimum SAU203 has the potential for the removal as well as detoxification of nickel from contaminated environments in general and particularly from the chromite mining areas of Odhisa, India.

Keywords: nickel, fungal biosorption, Penicillium simplicissimum SAU203, Indian chromite mines, mining overburden

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110 Recognition of a Thinly Bedded Distal Turbidite: A Case Study from a Proterozoic Delta System, Chaossa Formation, Simla Group, Western Lesser Himalaya, India

Authors: Priyanka Mazumdar, Ananya Mukhopadhyay

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A lot of progress has been achieved in the research of turbidites during the last decades. However, their relationship to delta systems still deserves further attention. This paper addresses example of fine grained turbidite from a pro-deltaic deposit of a Proterozoic mixed energy delta system exposed along Chaossa-Baliana river section of the Chaossa Formation of the Simla Basin. Lithostratigraphic analysis of the Chaossa Formation reveals three major facies associations (prodelta deposit-FA1, delta slope deposit-FA2 and delta front deposit-FA3) based on lithofacies types, petrography and sedimentary structures. Detailed process-based facies and paleoenvironmental analysis of the study area have led to identification of more than150 m thick coarsening-upwards deltaic successions composed of fine grained turbidites overlain by delta slope deposits. Erosional features are locally common at the base of turbidite beds and still more widespread at the top. The complete sequence has eight sub-divisions that are here termed T1 to T8. The basal subdivision (T1) comprises a massive graded unit with a sharp, scoured base, internal parallel-lamination and cross-lamination. The overlying sequence shows textural and compositional grading through alternating silt and mud laminae (T2). T2 is overlying by T3 which is characterized by climbing ripple and cross lamination. Parallel laminae are the predominant facies attributes of T4 which caps the T3 unit. T5 has a loaded scour base and is mainly characterized laminated silt. The topmost three divisions, graded mud (T6), ungraded mud (T7) and laminated mud (T8). The proposed sequence is analogous to the Bouma (1962) structural scheme for sandy turbidites. Repetition of partial sequences represents deposition from different stages of evolution of a large, muddy, turbidity flow. Detailed facies analysis of the study area reveals that the sediments of the turbidites developed during normal regression at the stage of stable or marginally rising sea level. Thin-bedded turbidites were deposited predominantly by turbidity currents in the relatively shallower part of the Simla basin. The fine-grained turbidites are developed by resedimentation of delta-front sands and slumping of upper pro-delta muds.

Keywords: turbidites, prodelta, proterozoic, Simla Basin, Bouma sequence

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109 Characterization of a Lipolytic Enzyme of Pseudomonas nitroreducens Isolated from Mealworm's Gut

Authors: Jung-En Kuan, Whei-Fen Wu

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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

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108 A Green Process for Drop-In Liquid Fuels from Carbon Dioxide, Water, and Solar Energy

Authors: Jian Yu

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Carbo dioxide (CO2) from fossil fuel combustion is a prime green-house gas emission. It can be mitigated by microalgae through conventional photosynthesis. The algal oil is a feedstock of biodiesel, a carbon neutral liquid fuel for transportation. The conventional CO2 fixation, however, is quite slow and affected by the intermittent solar irradiation. It is also a technical challenge to reform the bio-oil into a drop-in liquid fuel that can be directly used in the modern combustion engines with expected performance. Here, an artificial photosynthesis system is presented to produce a biopolyester and liquid fuels from CO2, water, and solar power. In this green process, solar energy is captured using photovoltaic modules and converted into hydrogen as a stable energy source via water electrolysis. The solar hydrogen is then used to fix CO2 by Cupriavidus necator, a hydrogen-oxidizing bacterium. Under the autotrophic conditions, CO2 was reduced to glyceraldehyde-3-phosphate (G3P) that is further utilized for cell growth and biosynthesis of polyhydroxybutyrate (PHB). The maximum cell growth rate reached 10.1 g L-1 day-1, about 25 times faster than that of a typical bio-oil-producing microalga (Neochloris Oleoabundans) under stable indoor conditions. With nitrogen nutrient limitation, a large portion of the reduced carbon is stored in PHB (C4H6O2)n, accounting for 50-60% of dry cell mass. PHB is a biodegradable thermoplastic that can find a variety of environmentally friendly applications. It is also a platform material from which small chemicals can be derived. At a high temperature (240 - 290 oC), the biopolyester is degraded into crotonic acid (C4H6O2). On a solid phosphoric acid catalyst, PHB is deoxygenated via decarboxylation into a hydrocarbon oil (C6-C18) at 240 oC or so. Aromatics and alkenes are the major compounds, depending on the reaction conditions. A gasoline-grade liquid fuel (77 wt% oil) and a biodiesel-grade fuel (23 wt% oil) were obtained from the hydrocarbon oil via distillation. The formation routes of hydrocarbon oil from crotonic acid, the major PHB degradation intermediate, are revealed and discussed. This work shows a novel green process from which biodegradable plastics and high-grade liquid fuels can be directly produced from carbon dioxide, water and solar power. The productivity of the green polyester (5.3 g L-1 d-1) is much higher than that of microalgal oil (0.13 g L-1 d-1). Other technical merits of the new green process may include continuous operation under intermittent solar irradiation and convenient scale up in outdoor.

Keywords: bioplastics, carbon dioxide fixation, drop-in liquid fuels, green process

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107 Effect of Several Soil Amendments on Water Quality in Mine Soils: Leaching Columns

Authors: Carmela Monterroso, Marc Romero-Estonllo, Carlos Pascual, Beatriz Rodríguez-Garrido

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The mobilization of heavy metals from polluted soils causes their transfer to natural waters, with consequences for ecosystems and human health. Phytostabilization techniques are applied to reduce this mobility, through the establishment of a vegetal cover and the application of soil amendments. In this work, the capacity of different organic amendments to improve water quality and reduce the mobility of metals in mine-tailings was evaluated. A field pilot test was carried out with leaching columns installed on an old Cu mine ore (NW of Spain) which forms part of the PhytoSUDOE network of phytomanaged contaminated field sites (PhytoSUDOE/ Phy2SUDOE Projects (SOE1/P5/E0189 and SOE4/P5/E1021)). Ten columns (1 meter high by 25 cm in diameter) were packed with untreated mine tailings (control) or those treated with organic amendments. Applied amendments were based on different combinations of municipal wastes, bark chippings, biomass fly ash, and nanoparticles like aluminum oxides or ferrihydrite-type iron oxides. During the packing of the columns, rhizon-samplers were installed at different heights (10, 20, and 50 cm) from the top, and pore water samples were obtained by suction. Additionally, in each column, a bottom leachate sample was collected through a valve installed at the bottom of the column. After packing, the columns were sown with grasses. Water samples were analyzed for: pH and redox potential, using combined electrodes; salinity by conductivity meter: bicarbonate by titration, sulfate, nitrate, and chloride, by ion chromatography (Dionex 2000); phosphate by colorimetry with ammonium molybdate/ascorbic acid; Ca, Mg, Fe, Al, Mn, Zn, Cu, Cd, and Pb by flame atomic absorption/emission spectrometry (Perkin Elmer). Porewater and leachate from the control columns (packed with unamended mine tailings) were extremely acidic and had a high concentration of Al, Fe, and Cu. In these columns, no plant development was observed. The application of organic amendments improved soil conditions, which allowed the establishment of a dense cover of grasses in the rest of the columns. The combined effect of soil amendment and plant growth had a positive impact on water quality and reduced mobility of aluminum and heavy metals.

Keywords: leaching, organic amendments, phytostabilization, polluted soils

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106 Association between Organophosphate Pesticides Exposure and Cognitive Behavior in Taipei Children

Authors: Meng-Ying Chiu, Yu-Fang Huang, Pei-Wei Wang, Yi-Ru Wang, Yi-Shuan Shao, Mei-Lien Chen

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Background: Organophosphate pesticides (OPs) are the most heavily used pesticides in agriculture in Taiwan. Therefore, they are commonly detected in general public including pregnant women and children. These compounds are proven endocrine disrupters that may affect the neural development in humans. The aim of this study is to assess the OPs exposure of children in 2 years of age and to examine the association between the exposure concentrations and neurodevelopmental effects in children. Methods: In a prospective cohort of 280 mother-child pairs, urine samples of prenatal and postnatal were collected from each participant and analyzed for metabolites of OPs by using gas chromatography-mass spectrometry. Six analytes were measured including dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphate (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). This study created a combined concentration measure for dimethyl compounds (DMs) consisting of the three dimethyl metabolites (DMP, DMTP, and DMDTP), for diethyl compounds (DEs) consisting of the three diethyl metabolites (DEP, DETP, and DEDTP) and six dialkyl phosphate (DAPs). The Bayley Scales of Infant and Toddler Development (Bayley-III) was used to assess children's cognitive behavior at 2 years old. The association between OPs exposure and Bayley-III scale score was determined by using the Mann-Whitney U test. Results: The measurements of urine samples are still on-going. This preliminary data are the report of 56 children aged 2 from the cohort. The detection rates for DMP, DMTP, DMDTP, DEP, DETP, and DEDTP are 80.4%, 69.6%, 64.3%, 64.3%, 62.5%, and 75%, respectively. After adjusting the creatinine concentrations of urine, the median (nmol/g creatinine) of urinary DMP, DMTP, DMDTP, DEP, DETP, DEDTP, DMs, DEs, and DAPs are 153.14, 53.32, 52.13, 19.24, 141.65, 192.17, 308.8, 311.6, and 702.11, respectively. The concentrations of urine are considerably higher than that in other countries. Children’s cognitive behavior was used three scales for Bayley-III, including cognitive, language and motor. In Mann-Whitney U test, the higher levels of DEs had significantly lower motor score (p=0.037), but no significant association was found between the OPs exposure levels and the score of either cognitive or language. Conclusion: The limited sample size suggests that Taipei children are commonly exposed to OPs and OPs exposure might affect the cognitive behavior of young children. This report will present more data to verify the results. The predictors of OPs concentrations, such as dietary pattern will also be included.

Keywords: biomonitoring, children, neurodevelopment, organophosphate pesticides exposure

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105 A World Map of Seabed Sediment Based on 50 Years of Knowledge

Authors: T. Garlan, I. Gabelotaud, S. Lucas, E. Marchès

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Production of a global sedimentological seabed map has been initiated in 1995 to provide the necessary tool for searches of aircraft and boats lost at sea, to give sedimentary information for nautical charts, and to provide input data for acoustic propagation modelling. This original approach had already been initiated one century ago when the French hydrographic service and the University of Nancy had produced maps of the distribution of marine sediments of the French coasts and then sediment maps of the continental shelves of Europe and North America. The current map of the sediment of oceans presented was initiated with a UNESCO's general map of the deep ocean floor. This map was adapted using a unique sediment classification to present all types of sediments: from beaches to the deep seabed and from glacial deposits to tropical sediments. In order to allow good visualization and to be adapted to the different applications, only the granularity of sediments is represented. The published seabed maps are studied, if they present an interest, the nature of the seabed is extracted from them, the sediment classification is transcribed and the resulted map is integrated in the world map. Data come also from interpretations of Multibeam Echo Sounder (MES) imagery of large hydrographic surveys of deep-ocean. These allow a very high-quality mapping of areas that until then were represented as homogeneous. The third and principal source of data comes from the integration of regional maps produced specifically for this project. These regional maps are carried out using all the bathymetric and sedimentary data of a region. This step makes it possible to produce a regional synthesis map, with the realization of generalizations in the case of over-precise data. 86 regional maps of the Atlantic Ocean, the Mediterranean Sea, and the Indian Ocean have been produced and integrated into the world sedimentary map. This work is permanent and permits a digital version every two years, with the integration of some new maps. This article describes the choices made in terms of sediment classification, the scale of source data and the zonation of the variability of the quality. This map is the final step in a system comprising the Shom Sedimentary Database, enriched by more than one million punctual and surface items of data, and four series of coastal seabed maps at 1:10,000, 1:50,000, 1:200,000 and 1:1,000,000. This step by step approach makes it possible to take into account the progresses in knowledge made in the field of seabed characterization during the last decades. Thus, the arrival of new classification systems for seafloor has improved the recent seabed maps, and the compilation of these new maps with those previously published allows a gradual enrichment of the world sedimentary map. But there is still a lot of work to enhance some regions, which are still based on data acquired more than half a century ago.

Keywords: marine sedimentology, seabed map, sediment classification, world ocean

Procedia PDF Downloads 219
104 Identifying Controlling Factors for the Evolution of Shallow Groundwater Chemistry of Ellala Catchment, Northern Ethiopia

Authors: Grmay Kassa Brhane, Hailemariam Siyum Mekonen

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This study was designed to identify the hydrogeochemical and anthropogenic processes controlling the evaluation of groundwater chemistry in the Ellala catchment which covers about 296.5 km2 areal extent. The chemical analysis revealed that the major ions in the groundwater are Ca2+, Mg2+, Na+, and K+ (cations) and HCO3-, PO43-, Cl-, NO3-, and SO42-(anions). Most of the groundwater samples (68.42%) revealed that the groundwater in the catchment is non-alkaline. In addition to the contribution of aquifer material, the solid materials and liquid wastes discharged from different sources can be the main sources of pH and EC in the groundwater. It is observed that the EC of the groundwater is fairly correlated with the DTS. This indicates that high mineralized water is more conductor than water with low concentration. The degree of salinity of the groundwater increases along the groundwater flow path from East to West; then, areas surrounding Mekelle City are highly saline due to the liquid and solid wastes discharged from the city and the industries. The groundwater facies in the catchment are predominated with calcium, magnesium, and bicarbonate which are labeled as Ca-Mg-HCO3 and Mg-Ca-HCO3. The main geochemical process controlling the evolution of the groundwater chemistry in the catchment is rock-water interaction, particularly carbonate dissolution. Due to the clay layer in the aquifer, the reverse is ion exchange. Non-significant silicate weathering and halite dissolution also contribute to the evolution of groundwater chemistry in the catchment. The groundwater in the catchment is dominated by the meteoritic origin although it needs further groundwater chemistry study with isotope dating analysis. The groundwater is under-saturated with calcite, dolomite, and aragonite minerals; hence, the more these minerals encounter the groundwater, the more the minerals dissolve. The main source of calcium and magnesium in groundwater is the dissolution of carbonate minerals (calcite and dolomite) since carbonate rocks are the dominant aquifer materials in the catchment. In addition to this, the weathering of dolerite rock is a possible source of magnesium ions. The relatively higher concentration of sodium over chloride indicates that the source of sodium-ion is reverse ion exchange and/or weathering of sodium-bearing materials, such as shale and dolerite rather than halite dissolution. High concentration of phosphate, nitrate, and chloride in the groundwater is the main anthropogenic source that needs treatment, quality control, and management in the catchment. From the Base Exchange Index Analysis, it is possible to understand that, in the catchment, the groundwater is dominated by the meteoritic origin, although it needs further groundwater chemistry study with isotope dating analysis.

Keywords: Ellala catchment, factor, chemistry, geochemical, groundwater

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103 Fatty Acid Translocase (Cd36), Energy Substrate Utilization, and Insulin Signaling in Brown Adipose Tissue in Spontaneously Hypertensive Rats

Authors: Michal Pravenec, Miroslava Simakova, Jan Silhavy

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Brown adipose tissue (BAT) plays an important role in lipid and glucose metabolism in rodents and possibly also in humans. Recently, using systems genetics approach in the BAT from BXH/HXB recombinant inbred strains, derived from the SHR (spontaneously hypertensive rat) and BN (Brown Norway) progenitors, we identified Cd36 (fatty acid translocase) as the hub gene of co-expression module associated with BAT relative weight and function. An important aspect of BAT biology is to better understand the mechanisms regulating the uptake and utilization of fatty acids and glucose. Accordingly, BAT function in the SHR that harbors mutant nonfunctional Cd36 variant (hereafter referred to as SHR-Cd36⁻/⁻) was compared with SHR transgenic line expressing wild type Cd36 under control of a universal promoter (hereafter referred to as SHR-Cd36⁺/⁺). BAT was incubated in media containing insulin and 14C-U-glucose alone or 14C-U-glucose together with palmitate. Incorporation of glucose into BAT lipids was significantly higher in SHR-Cd36⁺/⁺ versus SHR-Cd36⁻/⁻ rats when incubation media contained glucose alone (SHR-Cd36⁻/⁻ 591 ± 75 vs. SHR-Cd36⁺/⁺ 1036 ± 135 nmol/gl./2h; P < 0.005). Adding palmitate into incubation media had no effect in SHR-Cd36⁻/⁻ rats but significantly reduced glucose incorporation into BAT lipids in SHR-Cd36⁺/⁺ (SHR-Cd36⁻/⁻ 543 ± 55 vs. SHR-Cd36⁺/⁺ 766 ± 75 nmol/gl./2h; P < 0.05 denotes significant Cd36 x palmitate interaction determined by two-way ANOVA). This Cd36-dependent reduced glucose uptake in SHR-Cd36⁺/⁺ BAT was likely secondary to increased palmitate incorporation and utilization due to the presence of wild type Cd36 fatty acid translocase in transgenic rats. This possibility is supported by increased incorporation of 14C-U-palmitate into BAT lipids in the presence of both palmitate and glucose in incubation media (palmitate alone: SHR-Cd36⁻/⁻ 870 ± 21 vs. SHR-Cd36⁺/⁺ 899 ± 42; glucose+palmitate: SHR-Cd36⁻/⁻ 899 ± 47 vs. SHR-Cd36⁺/⁺ 1460 ± 111 nmol/palm./2h; P < 0.05 denotes significant Cd36 x glucose interaction determined by two-way ANOVA). It is possible that addition of glucose into the incubation media increased palmitate incorporation into BAT lipids in SHR-Cd36⁺/⁺ rats because of glucose availability for glycerol phosphate production and increased triglyceride synthesis. These changes in glucose and palmitate incorporation into BAT lipids were associated with significant differential expression of Irs1, Irs2, Slc2a4 and Foxo1 genes involved in insulin signaling and glucose metabolism only in SHR-Cd36⁺/⁺ rats which suggests Cd36-dependent effects on insulin action. In conclusion, these results provide compelling evidence that Cd36 plays an important role in BAT insulin signaling and energy substrate utilization.

Keywords: brown adipose tissue, Cd36, energy substrate utilization, insulin signaling, spontaneously hypertensive rat

Procedia PDF Downloads 123
102 Comparative Histological, Immunohistochemical and Biochemical Study on the Effect of Vit. C, Vit. E, Gallic Acid and Silymarin on Carbon Tetrachloride Model of Liver Fibrosis in Rats

Authors: Safaa S. Hassan, Mohammed H. Elbakry, Safwat A. Mangoura, Zainab M. Omar

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Background: Liver fibrosis is the main reason for increased mortality in chronic liver disease. It has no standard treatment. Antioxidants from a variety of sources are capable of slowing or preventing oxidation of other molecules. Aim: to evaluate the hepatoprotective effect of vit. C, vit. E and gallic acid in comparison to silymarin in the rat model of carbon tetrachloride induced liver fibrosis and their possible mechanisms of action. Material& Methods: A total number of 60 adult male albino rats 160-200gm were divided into six equal groups; received subcutaneous (s.c) injection for 8 weeks. Group I: as control. Group II: received 1.5 mL/kg of CCL4 .Group III: CCL4 and co- treatment with silymarin 100mg/kg p.o. daily. Group IV: CCL4 and co-treatment with vit. C 50mg/kg p.o. daily. Group V: CCL4 and co-treatment with vit. E 200mg/kg. p.o. Group VI: CCL4 and co-treatment with Gallic acid 100mg/kg. p.o. daily. Liver was processed for histological and immunohistochemical examination. Levels of AST, ALT, ALP, reduced GSH, MDA, SOD and hydroxyproline concentration were measured and evaluated statistically. Results: Light and electron microscopic examination of liver of group II exhibited foci of altered cells with dense nuclei and vacuolated, granular cytoplasm, mononuclear cell infiltration in portal areas, profuse collagen fiber deposits were found around portal tract, more intense staining α-SMA-positive cells occupied most of the liver fibrosis tissue, electron lucent areas in the cytoplasm of the hepatocytes, margination of nuclear chromatin. Treatment by any of the antioxidants variably reduced the hepatic structural changes induced by CCL4. Biochemical analysis showed that carbon tetrachloride significantly increased the levels of serum AST, ALT, ALP, hepatic malondialdehyde and hydroxyproline content. Moreover, it decreased the activities of superoxide dismutase and glutathione. Treatment with silymarin, gallic acid, vit. C and vit. E decreased significantly the AST, ALT, and ALP levels in plasma, MDA and hydroxyproline and increased the activities of SOD and glutathione in liver tissue. The effect of administration of CCl4 was improved with the used antioxidants in variable degrees. The most efficient antioxidant was silymarin followed by gallic acid and vit. C then vit. E. It is possibly due to their antioxidant effect, free radical scavenging properties and the reduction of oxidant dependent activation and proliferation of HSCs. Conclusion: So these antioxidants can be a promising drugs candidate for ameliorating liver fibrosis better than the use of the drugs and their side effects.

Keywords: antioxidant, ccl4, gallic acid, liver fibrosis

Procedia PDF Downloads 255
101 Rice Tablet Poisoning in Iran

Authors: Somayeh Khanjani, Samaneh Nabavi, Shirin Jalili

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Aluminum phosphide (ALP) is an inorganic phosphide used to control insects and is a highly effective insecticide and rodenticide used frequently to protect stored grain. Acute poisoning with this compound is common in some countries including India and Iran, and is a serious health problem. In Iran it was known as "rice tablet", for its use to preserve rice. Two kinds of rice tablets one being herbal while other containing 3g aluminum phosphide (AlP) are available for use in Iranian households to protect stored food grains from pests and rodents. The toxicity of Aluminum phosphide is attributed to the liberation of phosphine gas in contact with water or weak acid and is the major cause of poisoning and deaths. Rice tablet (Aluminum Phosphid) poisoning may be associated with serious and sometimes incurable complications. In 61.3% of patients were shown uniform ingestion. Vomiting was the most common symptoms reported by 96.4% patients. Agitation was reported in 36.9% and felling of thirsty in 27.9 %. Although many complications such as Hypotension, Adult Respiratory Distress Syndrome (ARDS), Acute Renal Failure (ARF) AND Multi Organ Failure (MOF) were the common complications observed in these patients, but the most lethal complication was Cardiac Arrhythmias occurred in 36.9% of cases. Abdominal pain in 31.4% of the patients, nausea in 79.4% of the patients and 41.1% of the patients showed metabolic acidosis. Suicidal intention was the most common cause of poisoning leading to deaths in 18.6% of the patients. Aluminum phosphide can cause either elevation, decrease or no change in electrolytes, bicarbonate and blood glucose level. The possible mechanism for changes in blood glucose levels are complex and depend on the balance of factors which increase its concentration and those which reduce it. AlP poisoning has been postulated to stimulate cortisol which leads to increasing blood level of cortisol, also it may cause stimulation of glucagon, and Adrenaline secretion; in addition, it can inhibit insulin synthesis which may lead to hyperglycemia. Another suggested mechanism of hyperglycemia is rennin activity in some cases, an increase in magnesium level of plasma and that of tissues, and high phosphate level. Although hyperglycemia is most frequent in this poisoning and also is known as a marker of poor prognostic, hypoglycemia in aluminum phosphide poisoning is a rare finding which may be so dangerous. Patients showed sever hypotension and sever acidosis in addition to sever hypoglycemia. The presenting features of AlP intoxication are rapid onset of shock, severe metabolic acidosis, cardiac dysrhythmias and adult respiratory distress syndrome (ARDS).

Keywords: aluminum phosphide (ALP), rice tablet, poisoning, phosphine gas

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100 The Development of Noctiluca scintillans Algal Bloom in Coastal Waters of Muscat, Sulanate of Oman

Authors: Aysha Al Sha'aibi

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Algal blooms of the dinoflagellate species Noctiluca scintillans became frequent events in Omani waters. The current study aims at elucidating the abundance, size variation and observations on the feeding mechanism performed by this species during the winter bloom. An attempt was made, to relate observed biological parameters of the Noctiluca population to environmental factors. Field studies spanned the period from December 2014 to April 2015. Samples were collected from Bandar Rawdah (Muscat region) by Bongo nets, twice per week, from the surface and the integrated upper mixed layer. The measured environmental variables were: temperature, salinity, dissolved oxygen, chlorophyll a, turbidity, nitrite, phosphate, wind speed and rainfall. During the winter bloom (from December 2014 through February 2015), the abundance exhibited the highest concentration on 17 February (640.24×106 cell.L-1) in oblique samples and 83.9x103 cell.L-1 in surface samples, with a subsequent decline up to the end of April. The average number of food vacuoles inside Noctiluca cells was 1.5 per cell; the percentage of feeding Noctiluca compared to the entire population varied from 0.01% to 0.03%. Both the surface area of the Noctiluca symbionts (Pedinomonas noctilucae) and cell diameter were maximal in December. In oblique samples the highest average cell diameter and the surface area of symbiont algae were 751.7 µm and 179.2x103 µm2 respectively. In surface samples, highest average cell diameter and the surface area of symbionts were 760 µm and 284.05x103 µm2 respectively. No significant correlations were detected between Noctiluca’s biological parameters and environmental variables except for the correlation between cell diameter and chlorophyll a, also between symbiotic algae surface area and chlorophyll a. The high correlation of chlorophyll a was as a reason of endosymbiotic algae Pedinomonas noctilucae and green Noctiluca enhanced chlorophyll during bloom. All correlations among biological parameters were significant; they are perhaps one of major factors that mediating high growth rates, generating millions of cell per liter in a short time range. The results gained from this study will provide a beneficial background for understanding deeply the development of coastal algal blooms of Noctiluca scintillans. Moreover, results could be used in different applications related to marine environment.

Keywords: abundance, feeding activities, Noctiluca scintillans, Oman

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99 Selective Extraction of Lithium from Native Geothermal Brines Using Lithium-ion Sieves

Authors: Misagh Ghobadi, Rich Crane, Karen Hudson-Edwards, Clemens Vinzenz Ullmann

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Lithium is recognized as the critical energy metal of the 21st century, comparable in importance to coal in the 19th century and oil in the 20th century, often termed 'white gold'. Current global demand for lithium, estimated at 0.95-0.98 million metric tons (Mt) of lithium carbonate equivalent (LCE) annually in 2024, is projected to rise to 1.87 Mt by 2027 and 3.06 Mt by 2030. Despite anticipated short-term stability in supply and demand, meeting the forecasted 2030 demand will require the lithium industry to develop an additional capacity of 1.42 Mt of LCE annually, exceeding current planned and ongoing efforts. Brine resources constitute nearly 65% of global lithium reserves, underscoring the importance of exploring lithium recovery from underutilized sources, especially geothermal brines. However, conventional lithium extraction from brine deposits faces challenges due to its time-intensive process, low efficiency (30-50% lithium recovery), unsuitability for low lithium concentrations (<300 mg/l), and notable environmental impacts. Addressing these challenges, direct lithium extraction (DLE) methods have emerged as promising technologies capable of economically extracting lithium even from low-concentration brines (>50 mg/l) with high recovery rates (75-98%). However, most studies (70%) have predominantly focused on synthetic brines instead of native (natural/real), with limited application of these approaches in real-world case studies or industrial settings. This study aims to bridge this gap by investigating a geothermal brine sample collected from a real case study site in the UK. A Mn-based lithium-ion sieve (LIS) adsorbent was synthesized and employed to selectively extract lithium from the sample brine. Adsorbents with a Li:Mn molar ratio of 1:1 demonstrated superior lithium selectivity and adsorption capacity. Furthermore, the pristine Mn-based adsorbent was modified through transition metals doping, resulting in enhanced lithium selectivity and adsorption capacity. The modified adsorbent exhibited a higher separation factor for lithium over major co-existing cations such as Ca, Mg, Na, and K, with separation factors exceeding 200. The adsorption behaviour was well-described by the Langmuir model, indicating monolayer adsorption, and the kinetics followed a pseudo-second-order mechanism, suggesting chemisorption at the solid surface. Thermodynamically, negative ΔG° values and positive ΔH° and ΔS° values were observed, indicating the spontaneity and endothermic nature of the adsorption process.

Keywords: adsorption, critical minerals, DLE, geothermal brines, geochemistry, lithium, lithium-ion sieves

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98 High Acid-Stable α-Amylase Production by Milk in Liquid Culture

Authors: Shohei Matsuo, Saki Mikai, Hiroshi Morita

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Objectives: Shochu is a popular Japanese distilled spirits. In the production of shochu, the filamentous fungus Aspergillus kawachii has traditionally been used. A. kawachii produces two types of starch hydrolytic enzymes, α-amylase (enzymatic liquefaction) and glucoamylase (enzymatic saccharification). Liquid culture system is a relatively easy microorganism to ferment with relatively low cost of production compared for solid culture. In liquid culture system, acid-unstable α-amylase (α-A) was produced abundantly, but, acid-stable α-amylase (Aα-A) was not produced. Since there is high enzyme productivity, most in shochu brewing have been adopted by a solid culture method. In this study, therefore, we investigated production of Aα-A in liquid culture system. Materials and methods: Microorganism Aspergillus kawachii NBRC 4308 was used. The mold was cultured at 30 °C for 7~14 d to allow formation of conidiospores on slant agar medium. Liquid Culture System: A. kawachii was cultured in a 100 ml of following altered SLS medium: 1.0 g of rice flour, 0.1 g of K2HPO4, 0.1 g of KCl, 0.6 g of tryptone, 0.05 g of MgSO4・7H2O, 0.001 g of FeSO4・7H2O, 0.0003 g of ZnSO4・7H2O, 0.021 g of CaCl2, 0.33 of citric acid (pH 3.0). The pH of the medium was adjusted to the designated value with 10 % HCl solution. The cultivation was shaking at 30 °C and 200 rpm for 72 h. It was filtered to obtain a crude enzyme solution. Aα-A assay: The crude enzyme solution was analyzed. An acid-stable α-amylase activity was carried out using an α-amylase assay kit (Kikkoman Corporation, Noda, Japan). It was conducted after adding 9 ml of 100 mM acetate buffer (pH 3.0) to 1 ml of the culture product supernatant and acid treatment at 37°C for 1 h. One unit of a-amylase activity was defined as the amount of enzyme that yielded 1 mmol of 2-chloro-4-nitrophenyl 6-azide-6-deoxy-b-maltopentaoside (CNP) per minute. Results and Conclusion: We experimented with co-culture of A. kawachii and lactobacillus in order to get control of pH in altered SLS medium. However, high production of acid-stable α-amylase was not obtained. We experimented with yoghurt or milk made an addition to liquid culture. The result indicated that high production of acid-stable α-amylase (964 U/g-substrate) was obtained when milk made an addition to liquid culture. Phosphate concentration in the liquid medium was a major cause of increased acid-stable α-amylase activity. In liquid culture, acid-stable α-amylase activity was enhanced by milk, but Fats and oils in the milk were oxidized. In addition, Tryptone is not approved as a food additive in Japan. Thus, alter SLS medium added to skim milk excepting for the fats and oils in the milk instead of tryptone. The result indicated that high production of acid-stable α-amylase was obtained with the same effect as milk.

Keywords: acid-stable α-amylase, liquid culture, milk, shochu

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97 DPAGT1 Inhibitors: Discovery of Anti-Metastatic Drugs

Authors: Michio Kurosu

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Alterations in glycosylation not only directly impact cell growth and survival but also facilitate tumor-induced immunomodulation and eventual metastasis. Identification of cell type-specific glycoconjugates (tumor markers) has led to the discovery of new assay systems for certain cancers via immunodetection reagents. N- and O-linked glycans are the most abundant forms of glycoproteins. Recent studies of cancer immunotherapy are based on the immunogenicity of truncated O-glycan chains (e.g., Tn, sTn, T, and sLea/x). The prevalence of N-linked glycan changes in the development of tumor cells is known; however, therapeutic antibodies against N-glycans have not yet been developed. This is due to the lack of specificity of N-linked glycans between normal/healthy and cancer cells. Abnormal branching of N-linked glycans has been observed, particularly in solid cancer cells. While the discovery of drug-like glycosyltransferase inhibitors that block the biosynthesis of specific branching has a very low likelihood of success, altered glycosylation levels can be exploited by suppressing N-glycan biosynthesis through the inhibition of dolichyl-phosphate N-acetylglucosaminephosphotransferase1 (DPAGT1) activity. Inhibition of DPAGT1 function leads to changes of O-glycosylation on proteins associated with mitochondria and zinc finger binding proteins (indirect effects). On the basis of dynamic crosstalk between DPAGT1 and Snail/Slung/ZEB1 (a family of transcription factors that promote the repression of the adhesion molecules), we have developed pharmacologically acceptable selective DPAGT1 inhibitors. Tunicamycin kills a wide range of cancer and healthy cells in a non-selective manner. In sharp contrast, our DPAGT1 inhibitors display strong cytostatic effects against 16 solid cancers, which require the overexpression of DPAGT1 in their progression but do not affect the cell viability of healthy cells. The identified DPAGT1 inhibitors possess impressive anti-metastatic ability in various solid cancer cell lines and induce their mitochondrial structural changes, resulting in apoptosis. A prototype DPAGT1 inhibitor, APPB has already been proven to shrink solid tumors (e.g., pancreatic cancers, triple-negative breast cancers) in vivo while suppressing metastases and has strong synergistic effects when combined with current cytotoxic drugs (e.g., paclitaxel). At this conference, our discovery of selective DPAGT1 inhibitors with drug-like properties and proof-of-pharmaceutical concept studies of a novel DPAGT1 inhibitor are presented.

Keywords: DPAGT1 inhibitors, anti-metastatic drugs, natural product based drug designs, cytostatic effects

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96 Acoustic Radiation Pressure Detaches Myoblast from Culture Substrate by Assistance of Serum-Free Medium

Authors: Yuta Kurashina, Chikahiro Imashiro, Kiyoshi Ohnuma, Kenjiro Takemura

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Research objectives and goals: To realize clinical applications of regenerative medicine, a mass cell culture is highly required. In a conventional cell culture, trypsinization was employed for cell detachment. However, trypsinization causes proliferation decrease due to injury of cell membrane. In order to detach cells using an enzyme-free method, therefore, this study proposes a novel cell detachment method capable of detaching adherent cells using acoustic radiation pressure exposed to the dish by the assistance of serum-free medium with ITS liquid medium supplement. Methods used In order to generate acoustic radiation pressure, a piezoelectric ceramic plate was glued on a glass plate to configure an ultrasonic transducer. The glass plate and a chamber wall compose a chamber in which a culture dish is placed in glycerol. Glycerol transmits acoustic radiation pressure to adhered cells on the culture dish. To excite a resonance vibration of transducer, AC signal with 29-31 kHz (swept) and 150, 300, and 450 V was input to the transducer for 5 min. As a pretreatment to reduce cell adhesivity, serum-free medium with ITS liquid medium supplement was spread to the culture dish before exposed to acoustic radiation pressure. To evaluate the proposed cell detachment method, C2C12 myoblast cells (8.0 × 104 cells) were cultured on a ø35 culture dish for 48 hr, and then the medium was replaced with the serum-free medium with ITS liquid medium supplement for 24 hr. We replaced the medium with phosphate buffered saline and incubated cells for 10 min. After that, cells were exposed to the acoustic radiation pressure for 5 min. We also collected cells by using trypsinization as control. Cells collected by the proposed method and trypsinization were respectively reseeded in ø60 culture dishes and cultured for 24 hr. Then, the number of proliferated cells was counted. Results achieved: By a phase contrast microscope imaging, shrink of lamellipodia was observed before exposed to acoustic radiation pressure, and no cells remained on the culture dish after the exposed of acoustic radiation pressure. This result suggests that serum-free medium with ITS liquid inhibits adhesivity of cells and acoustic radiation pressure detaches cells from the dish. Moreover, the number of proliferated cells 24 hr after collected by the proposed method with 150 and 300 V is the same or more than that by trypsinization, i.e., cells were proliferated 15% higher with the proposed method using acoustic radiation pressure than with the traditional cell collecting method of trypsinization. These results proved that cells were able to be collected by using the appropriate exposure of acoustic radiation pressure. Conclusions: This study proposed a cell detachment method using acoustic radiation pressure by the assistance of serum-free medium. The proposed method provides an enzyme-free cell detachment method so that it may be used in future clinical applications instead of trypsinization.

Keywords: acoustic radiation pressure, cell detachment, enzyme free, ultrasonic transducer

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