Search results for: liquid cell
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 5454

Search results for: liquid cell

1314 1D PIC Simulation of Cold Plasma Electrostatic Waves beyond Wave-Breaking Limit

Authors: Prabal Singh Verma

Abstract:

Electrostatic Waves in plasma have emerged as a new source for the acceleration of charged particles. The accelerated particles have a wide range of applications, for example in cancer therapy to cutting and melting of hard materials. The maximum acceleration can only be achieved when the amplitude of the plasma wave stays below a critical limit known as wave-breaking amplitude. Beyond this limit amplitude of the wave diminishes dramatically as the coherent energy of the wave starts to convert into random kinetic energy. In this work, spatiotemporal evolution of non-relativistic electrostatic waves in a cold plasma has been studied in the wave-breaking regime using a 1D particle-in-cell simulation (PIC). It is found that plasma gets heated after the wave-breaking but a fraction of initial energy always remains with the remnant wave in the form of Bernstein-Greene-Kruskal (BGK) mode in warm plasma. Another interesting finding of this work is that the frequency of the resultant BGK wave is found be below electron plasma frequency which decreases with increasing initial amplitude and the acceleration mechanism after the wave-breaking is also found to be different from the previous work. In order to explain the results observed in the numerical experiments, a simplified theoretical model is constructed which exhibits a good agreement with the simulation. In conclusion, it is shown in this work that electrostatic waves get shower after the wave-breaking and a fraction of initial coherent energy always remains with remnant wave. These investigations have direct relevance in wakefield acceleration experiments.

Keywords: nonlinear plasma waves, longitudinal, wave-breaking, wake-field acceleration

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1313 Performance Comparison of Resource Allocation without Feedback in Wireless Body Area Networks by Various Pseudo Orthogonal Sequences

Authors: Ojin Kwon, Yong-Jin Yoon, Liu Xin, Zhang Hongbao

Abstract:

Wireless Body Area Network (WBAN) is a short-range wireless communication around human body for various applications such as wearable devices, entertainment, military, and especially medical devices. WBAN attracts the attention of continuous health monitoring system including diagnostic procedure, early detection of abnormal conditions, and prevention of emergency situations. Compared to cellular network, WBAN system is more difficult to control inter- and inner-cell interference due to the limited power, limited calculation capability, mobility of patient, and non-cooperation among WBANs. In this paper, we compare the performance of resource allocation scheme based on several Pseudo Orthogonal Codewords (POCs) to mitigate inter-WBAN interference. Previously, the POCs are widely exploited for a protocol sequence and optical orthogonal code. Each POCs have different properties of auto- and cross-correlation and spectral efficiency according to its construction of POCs. To identify different WBANs, several different pseudo orthogonal patterns based on POCs exploits for resource allocation of WBANs. By simulating these pseudo orthogonal resource allocations of WBANs on MATLAB, we obtain the performance of WBANs according to different POCs and can analyze and evaluate the suitability of POCs for the resource allocation in the WBANs system.

Keywords: wireless body area network, body sensor network, resource allocation without feedback, interference mitigation, pseudo orthogonal pattern

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1312 Assessment of Alteration in High Density Lipo Protein, Apolipoprotein A1, Serum Glutamic Pyruvic Transaminase and Serum Glutamic Oxaloacetic Transaminase in Oral Submucous Fibrosis Patients

Authors: Marina Lazar Chandy, N. Kannan, Rajendra Patil, Vinod Mathew, Ajmal Mohamed, P. K. Sreeja, Renju Jose

Abstract:

Introduction- Arecoline, a major constituent of arecanut has shown to have some effect on liver. The use of arecanut is found to be the most common etiological factor for the development of Oral Submucous fibrosis (O.S.M.F). The effect of arecanut usage on liver in patients with O.S.M.F needs to be assessed. Lipids play a role in structural maintenance of cell. Alterations of lipid profile were noted in cancer patients. O.S.M.F being a precancerous lesion can have some effect on the level of lipids in the body. Objectives: This study was done to assess the alterations in liver enzymes (Serum Glutamic Pyruvic Transaminase(S.G.P.T ,Serum Glutamic Oxaloacetic Transaminase(S.G.O.T)) and lipid metabolism (High Density Lipoprotien(H.D.L) and Apo Lipoprotien A1 (Apo A1)) in patients with O.S.M.F. Methods-130 patients were taken for the study,100 patients with O.S.M.F and 30 as control group without O.S.M.F. Fasting blood sugar levels were taken, centrifuged and analyzed for S.G.P.T,S.G.O.T, H.D.L and Apo A1 using semi automated spectrophotometer. Results: After statistical analysis, it was concluded that there is an elevation of levels of S.G.P.T, S.G.O.T, and decreased levels of H.D.L, Apo A1 for O.S.M.F group when compared with control group. With increased grade of O.S.M.F. and duration of habit, S.G.P.T. & S.G.O.T. increased whereas, H.D.L. & Apo A1 decreased. All the values were statistically significant at p<0.01.

Keywords: apolipoprotien A1, high density lipoprotien, oral submucous fibrosis, serum glutamic oxaloacetic transaminase

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1311 Comparison Between Partial Thickness Skin Graft Harvesting From Scalp and Lower Limb for Scalp Defect

Authors: Mehrdad Taghipour, Mina Rostami, Mahdi Eskandarlou

Abstract:

Partial-thickness skin graft is the cornerstone for scalp defect repair. Given the potential side effects following harvesting from these sites, this study aimed to compare the outcomes of graft harvesting from scalp and lower limb. This clinical trial was conducted among a sample number of 40 partial thickness graft candidates (20 case and 20 control group) with scalp defect presenting to Plastic Surgery Clinic at Besat Hospital, Hamadan, Iran during 2018-2019. Sampling was done by simple randomization using random digit table. The donor site in case group and control group was scalp and lower limb respectively. Overall, 28 patients (70%) were male and 12 (30%) were female. Basal cell carcinoma (BCC) and trauma were the most common etiology for the defects. There was a statistically meaningful relationship between two groups regarding the etiology of defect (P=0.02). The mean diameter of defect was 24.28±45.37 mm for all of the patients. The difference between diameters of defect in both groups were statistically meaningful while no such difference between graft diameters was seen. The graft “Take” was completely successful in both groups according to evaluations. The level of postoperative pain was lower in the case group compared to the control according to VAS scale and the satisfaction was higher in them per Likert scale. Scalp can safely be used as donor site for skin graft to be used for scalp defects associated with better results and lower complication rates compared to other donor sites.

Keywords: donor site, graft, scalp, partial thickness

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1310 Correlation of P53 Gene Expression With Serum Alanine Transaminase Levels and Hepatitis B Viral Load in Cirrhosis and Hepatocellular Carcinoma Patients

Authors: Umme Shahera, Saifullah Munshi, Munira Jahan, Afzalun Nessa, Shahinul Alam, Shahina Tabassum

Abstract:

The development of HCC is a multi-stage process. Several extrinsic factors, such as aflatoxin, HBV, nutrition, alcohol, and trace elements are thought to initiate or/and promote the hepatocarcinogenesis. Alteration of p53 status is an important intrinsic factor in this process as p53 is essential for preventing inappropriate cell proliferation and maintaining genome integrity following genotoxic stress. This study was designed to assess the correlation of p53 gene expression with HBV-DNA and serum Alanine transaminase (ALT) in patients with cirrhosis and HCC. The study was conducted among 60 patients. The study population were divided into four groups (15 in each groups)-HBV positive cirrhosis, HBV negative cirrhosis, HBV positive HCC and HBV negative HCC. Expression of p53 gene was observed using real time PCR. P53 gene expressions in the above mentioned groups were correlated with serum ALT level and HBV viral load. p53 gene was significantly higher in HBV-positive patients with HCC than HBV-positive cirrhosis. Similarly, the expression of p53 was significantly higher in HBV-positive HCC than HBV-negative HCC patients. However, the expression of p53 was reduced in HBV-positive cirrhosis in comparison with HBV-negative cirrhosis. P53 gene expression in liver was not correlated with the serum levels of ALT in any of the study groups. HBV- DNA load also did not correlated with p53 gene expression in HBV positive HCC and HBV positive cirrhosis patients. This study shows that there was no significant change with the expression of p53 gene in any of the study groups with ALT level or viral load, though differential expression of p53 gene were observed in cirrhosis and HCC patients.

Keywords: P53, ALT, HBV-DNA, liver cirrhosis, hepatocellular carcinoma

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1309 Impact of Some Experimental Procedures on Behavioral Patterns and Physiological Traits of Rats

Authors: Amira, A. Goma, U. E. Mahrous

Abstract:

Welfare may be considered to be a subjective experience; it has a biological function that is related to the fitness and survival of the animal accordingly, researches have suggested that welfare is compromised when the animal's evolutionary fitness is reduced. This study was carried out to explain the effect of some managerial stressors as handling and restraint on behavioral patterns and biochemical parameters of rats. A total of 24 (12 males and 12 females) Sprague-Dawley rats (12 months and 150-180g) were allotted into 3 groups, handled group (4 male and 4 female), restrained group (4 male and 4 female) and control group (4 males and 4 females). The obtained results revealed that time spent feeding, drinking frequency, movement and cage exploration increased significantly in handled rats than other groups, while lying time and licking increased significantly in restrained rats than handled and controls. Moreover, social behavior decreased in both stressed groups than control. Triglycerides were significantly increased in handled rats than other groups, while total lipid, total protein and globulin significantly increased in both treated groups than control. Corticosterone increased in restrained and handled rats than control ones. Moreover, there was an increment in packed cell volume significantly in restrained rats than others. These deducted that if we want to study the effect of stress on animal welfare it is necessary to study the effect of such stressors on animal’s behavior and physiological responses.

Keywords: handling, restraint, welfare, rat, behavior, physiology

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1308 Effect of Molecular Weight Distribution on Toughening Performance of Polybutadiene in Polystyrene

Authors: Mohamad Mohsen Yavarizadeh

Abstract:

Polystyrene (PS) and related homopolymers are brittle materials that typically fail in tensile tests at very low strains. These polymers can be toughened by the addition of rubbery particles which initiate a large number of crazes that produce substantial plastic strain at relatively low stresses. Considerable energy is dissipated in the formation of these crazes, producing a relatively tough material that shows an impact toughness of more than 5 times of pure PS. While cross linking of rubbery phase is necessary in aforementioned mechanism of toughening, another mechanism of toughening was also introduced in which low molecular weight liquid rubbers can also toughen PS when dispersed in the form of small pools in the glassy matrix without any cross linking. However, this new mechanism which is based on local plasticization, fails to act properly at high strain rate deformations, i.e. impact tests. In this work, the idea of combination of these two mechanisms was tried. To do so, Polybutadiene rubbers (PB) with bimodal distribution of molecular weight were prepared in which, comparable fractions of very high and very low molecular weight rubbers were mixed. Incorporation of these materials in PS matrix in a reactive process resulted in more significant increases in toughness of PS. In other words, although low molecular weight PB is ineffective in high strain rate impact test by itself, it showed a significant synergistic effect when combined with high molecular weight PB. Surprisingly, incorporation of just 10% of low molecular weight PB doubled the impact toughness of regular high impact PS (HIPS). It was observed that most of rubbery particles could initiate crazes. The effectiveness of low molecular weight PB in impact test was attributed to low strain rate deformation of each individual craze as a result of producing a large number of crazes in this material. In other words, high molecular weight PB chains make it possible to have an appropriate dispersion of rubbery phase in order to create a large number of crazes in the PS matrix and consequently decrease the velocity of each craze. Low molecular weight PB, in turn, would have enough time to locally plasticize craze fibrils and enhance the energy dissipation.

Keywords: molecular weight distribution, polystyrene, toughness, homopolymer

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1307 Cryopreservation of Ring-Necked Pheasant (Phasianus colchicus) Semen for Establishing Cryobank

Authors: Rida Pervaiz, Bushra Allah Rakha, Muhammad Sajjad Ansari, Shamim Akhter, Kainat Waseem, Sumiyyah Zuha, Tooba Javed

Abstract:

Ring-necked pheasant (Phasianus colchicus) belongs to order Galliformes and family Phasianidae. It has been recognized as the most hunted bird due to its attractive colorful appearance and meat. Loss of habitat and hunting pressure has caused population fluctuations in the native range. Under these circumstances, this species can be conserved by employing ex-situ in vitro conservation techniques. Captive breeding, in combination with semen cryobanking is the most appropriate option to conserve/propagate this species without deteriorating the genetic diversity. Cryopreservation protocols of adequate efficiency are necessary to establish semen cryobanking for a species. Therefore, present study was designed to devise an efficient extender for cryopreservation of ring-necked pheasant semen. For this purpose, a range of extenders (Beltsville Poultry, red fowl, Lake, EK, Tselutin Poultry and Chicken semen extenders) were evaluated for cryopreservation of ring-necked pheasant semen. Semen collected from 10 cocks, diluted in the Beltsville Poultry (BPSE), Red Fowl (RFE), Lake (LE), EK (EKE), Tselutin Poultry (TPE) and Chicken Semen (CSE) extenders and cryopreserved. Glycerol (10%) was added to semen at 4°C, equilibrated for 10 min, filled in 0.5 mL French straws, kept over liquid nitrogen vapors for 10 min, cryopreserved in LN2 and stored. Sperm motility (%), viability (%), live/dead ratio (%), plasma membrane (%) and DNA Integrity (%) were evaluated at post-dilution, post-cooling, post-equilibration and post-thawing stage of cryopreservation. Sperm motility (83.8 ± 3.1; 81.3 ± 3.8; 73.8 ± 2.4; 62.5 ± 1.4), viability (79.0 ± 1.7; 75.5 ± 1.6; 69.5 ± 2.3; 65.5 ± 2.4), live/dead ratio (80.5 ± 5.7; 77.3 ± 4.9; 76.0 ± 2.7; 68.3 ± 2.3), plasma membrane (74.5 ± 2.9; 73.8 ± 3.4; 71.3 ± 2.3; 75.0 ± 3.4) and DNA integrity (78.3 ± 1.7; 73.0 ± 1.2; 68.0 ± 2.0; 63.0 ± 2.5) at all four stages of cryopreservation were recorded higher (P < 0.05) in red fowl extender compared to all experimental extenders. It is concluded that red fowl extender is the best extender for cryopreservation of ring-necked pheasant semen and can be used in establishing cryobank for ex situ conservation.

Keywords: ring-necked pheasant; extenders; cryopreservation; semen quality; DNA integrity

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1306 Antioxidant Juice Prevents UV- Induced Skin Damage in Rats

Authors: S. P. Gomes, D. C. Goncalves, E. Ribeiro, M. C. L. Seelaender

Abstract:

Skin is susceptible to photo damage induced by exposure to sunlight, or ultraviolet (UV) radiation, which induces breakdown of extracellular matrix, DNA degradation, skin cell lesion and apoptosis, and development of cancer. Phytonutrients demonstrate protective effects against UV damage. The purpose of this study was evaluating the effect of an antioxidant juice (AJ) contaning Brazilian natural products upon skin damage. The juice was produced by Metabolics®. Male Wistar rats were divided in 4 groups: Animals receiving the antioxidant juice (AJ): orange, carrot, honey, tomato extract, avocado, ginger and camu-camu (Brazilian fruit, a major source of vitamin C) ad libitum for 21 days; or water (C), subdivided in groups exposed or not to UV radiation for 2 non consecutive days, during five hours each day, after 15 days of juice supplementation. On the 22nd day, rats were killed by decapitation and epithelium samples from the dorsal skin removed, fixed in bouin and embedded in paraffin. The sections were stained with hematoxylin and eosin or mallory and picrosirius red. Isolated DNA was submitted to electrophoresis (1.8% agarose gel, 0.5% ethidium bromide). UV radiation significantly induced sunburn of superficial epithelial cells of C, AJ treatment reduced this effect. Collagen changes were observed in UV groups, yet AJ treatment prevented collagen degradation. UV radiation induced significant DNA degradation, in C, which was prevented by AJ treatment. The antioxidant juice consumed chronically protected against acute skin damage.

Keywords: nutraceuticals, antioxidants, photoprotection, uv radiation

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1305 Docking, Pharmacophore Modeling and 3d QSAR Studies on Some Novel HDAC Inhibitors with Heterocyclic Linker

Authors: Harish Rajak, Preeti Patel

Abstract:

The application of histone deacetylase inhibitors is a well-known strategy in prevention of cancer which shows acceptable preclinical antitumor activity due to its ability of growth inhibition and apoptosis induction of cancer cell. Molecular docking were performed using Histone Deacetylase protein (PDB ID:1t69) and prepared series of hydroxamic acid based HDACIs. On the basis of docking study, it was predicted that compound 1 has significant binding interaction with HDAC protein and three hydrogen bond interactions takes place, which are essential for antitumor activity. On docking, most of the compounds exhibited better glide score values between -8 to -10 which is close to the glide score value of suberoylanilide hydroxamic acid. The pharmacophore hypotheses were developed using e-pharmacophore script and phase module. The 3D-QSAR models provided a good correlation between predicted and actual anticancer activity. Best QSAR model showed Q2 (0.7974), R2 (0.9200) and standard deviation (0.2308). QSAR visualization maps suggest that hydrogen bond acceptor groups at carbonyl group of cap region and hydrophobic groups at ortho, meta, para position of R9 were favorable for HDAC inhibitory activity. We established structure activity correlation using docking, pharmacophore modeling and atom based 3D QSAR model for hydroxamic acid based HDACIs.

Keywords: HDACIs, QSAR, e-pharmacophore, docking, suberoylanilide hydroxamic acid

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1304 Synthesis, Characterization and Antibacterial Activity of Metalloporphyrins: Role of Central Metal Ion

Authors: Belete B. Beyene, Ayenew M. Mihirteu, Misganaw T. Ayana, Amogne W. Yibeltal

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Modification of synthetic porphyrins is one of the promising strategies in an attempt to get molecules with desired properties and applications. Here in, we report synthesis, photophysical characterization and antibacterial activity of 5, 10, 15, 20-tetrakis-(4- methoxy carbonyl phenyl) porphyrin M(II); where M = Co, Fe, Ni, Zn. Metallation of the ligand was confirmed by using UV–Vis spectroscopy and ESI-Ms measurement, in which the number of Q bands in absorption spectra of the ligand decreased from four to one or two as a result of metal insertion to the porphyrin core. The antibacterial activity study of the complexes toward two Gram-positive (Staphylococcus aureus (S. aureus) and Streptococcus pyogenes (s. pyogenes)) and two Gram-negative (Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae)) bacteria by disc diffusion method showed a promising inhibitory activity. The complexes exhibited highest activities at highest concentration and were better than the activity of free base ligand, the salts, and blank solution. This could be explained on the basis of Overton's concept of cell permeability and Tweed's Chelation theory. An increased lipo-solubility enhances the penetration of the complexes into the lipid membrane and interferes with the normal activities of the bacteria. Our study, therefore, showed that the growth inhibitory effect of these metalloporphyrins is generally in order of ZnTPPCOOMe > NiTPPCOOMe > CoTPPCOOMe> FeTPPCOOMe, which may be attributed to the better lipophilicity and binding of the complex with the cellular components.

Keywords: porphyrins, metalloporphyrins, spectral property, antibacterial activity, synthesis

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1303 Isolation, Characterization and Myogenic Differentiation of Synovial Mesenchymal Stem Cells

Authors: Fatma Y. Meligy

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Objectives: The objectives of this study aimed to isolate and characterize mesenchymal stem cells (MSCs) derived from synovial membrane. Then to assess the potentiality of myogenic differentiation of these isolated MSCs. Methods: The MSCs were isolated from synovial membrane by digestion method. Three adult rats were used. The 5 -azacytidine was added to the cultured cells for one day. The isolated cells and treated cells are assessed using immunoflouresence, flowcytometry, PCR and real time PCR. Results: The isolated stem cells showed morphological aspect of stem cells they showed strong positivity to CD44 and CD90 in immunoflouresence while in CD34 and CD45 showed negative reaction. The treated cells with 5-azacytidine was shown to have positive reaction for desmin. Flowcytometric analysis showed that synovial MSCs had strong positive percentage for CD44(%98)and CD90 (%97) and low percentage for CD34 & CD45 while the treated cells showed positive percentage for myogenic marker myogenin (85%). As regard the PCR and Real time PCR, the treated cells showed positive reaction to the desmin primer. Conclusion: The adult MSCs were isolated successfully from synovial membrane and characterized with stem cell markers. The isolated cells could be differentiated in vitro into myogenic cells. These differentiated cells could be used in auto-replacement of diseased or traumatized muscle cells as a regenerative therapy for muscle disorders and trauma.

Keywords: mesenchymal stem cells, synovial membrane, myogenic differentiation

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1302 Hydrometallurgical Recovery of Cobalt, Nickel, Lithium, and Manganese from Spent Lithium-Ion Batteries

Authors: E. K. Hardwick, L. B. Siwela, J. G. Falconer, M. E. Mathibela, W. Rolfe

Abstract:

Lithium-ion battery (LiB) demand has increased with the advancement in technologies. The applications include electric vehicles, cell phones, laptops, and many more devices. Typical components of the cathodes include lithium, cobalt, nickel, and manganese. Recycling the spent LiBs is necessary to reduce the ecological footprint of their production and use and to have a secondary source of valuable metals. A hydrometallurgical method was investigated for the recovery of cobalt and nickel from LiB cathodes. The cathodes were leached using a chloride solution. Ion exchange was then used to recover the chloro-complexes of the metals. The aim of the research was to determine the efficiency of a chloride leach, as well as ion exchange operating capacities that can be achieved for LiB recycling, and to establish the optimal operating conditions (ideal pH, temperature, leachate and eluant, flowrate, and reagent concentrations) for the recovery of the cathode metals. It was found that the leaching of the cathodes could be hindered by the formation of refractory metal oxides of cathode components. A reducing agent was necessary to improve the leaching rate and efficiency. Leaching was achieved using various chloride-containing solutions. The chloro-complexes were absorbed by the ion exchange resin and eluted to produce concentrated cobalt, nickel, lithium, and manganese streams. Chromatographic separation of these elements was achieved. Further work is currently underway to determine the optimal operating conditions for the recovery by ion exchange.

Keywords: cobalt, ion exchange, leachate formation, lithium-ion batteries, manganese, nickel

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1301 Engineered Reactor Components for Durable Iron Flow Battery

Authors: Anna Ivanovskaya, Alexandra E. L. Overland, Swetha Chandrasekaran, Buddhinie S. Jayathilake

Abstract:

Iron-based redox flow batteries (IRFB) are promising for grid-scale storage because of their low-cost and environmental safety. Earth-abundant iron can enable affordable grid-storage to meet DOE’s target material cost <$20/kWh and levelized cost for storage $0.05/kWh. In conventional redox flow batteries, energy is stored in external electrolyte tanks and electrolytes are circulated through the cell units to achieve electrochemical energy conversions. However, IRFBs are hybrid battery systems where metallic iron deposition at the negative side of the battery controls the storage capacity. This adds complexity to the design of a porous structure of 3D-electrodes to achieve a desired high storage capacity. In addition, there is a need to control parasitic hydrogen evolution reaction which accompanies the metal deposition process, increases the pH, lowers the energy efficiency, and limits the durability. To achieve sustainable operation of IRFBs, electrolyte pH, which affects the solubility of reactants and the rate of parasitic reactions, needs to be dynamically readjusted. In the present study we explore the impact of complexing agents on maintaining solubility of the reactants and find the optimal electrolyte conditions and battery operating regime, which are specific for IRFBs with additives, and demonstrate the robust operation.

Keywords: flow battery, iron-based redox flow battery, IRFB, energy storage, electrochemistry

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1300 Development of Self Emulsifying Drug Delivery Systems (SEDDS) of Anticancer Agents Used in AYUSH System of Medicine for Improved Oral Bioavailability Followed by Their Pharmacological Evaluation Using Biotechnological Techniques

Authors: Meenu Mehta, Munish Garg

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The use of oral anticancer drugs from AYUSH system of medicine is widely increased among the society due to their low cost, enhanced efficacy, increased patient preference, lack of inconveniences related to infusion and they provide an opportunity to develop chronic treatment regimens. However, oral delivery of these drugs usually laid down by the limited bioavailability of the drug, which is associated with a wide variation. As most of the cytotoxic agents have a narrow therapeutic window and are dosed at or near the maximum tolerated dose, a wide variability in the bioavailability can negatively affect treatment result. It is estimated that 40% of active substances are poorly soluble in water. The improvement of bio-availability of drugs with such properties presents one of the greatest challenges in drug formulations. There are several techniques reported in literature. Among all these Self Emulsifying Drug Delivery System (SEDDS) has gained more attention due to enhanced oral bio-availability enabling a reduction in dose. Thus, SEDDS anticancer drugs will have the increased bioavailability and efficacy. These dosage form will provide societal benefit in a cost-effective manner as compared to other oral dosage forms. Present study reflects on the formulation strategies as SEDDS for oral anticancer agents of AYUSH system for enhanced bioavailability with proven efficacy by cancer cell lines.

Keywords: anticancer agents, AYUSH system, bioavailability, SEDDS

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1299 Extracellular Enzymes from Halophilic Bacteria with Potential in Agricultural Secondary Flow Recovery Products

Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi

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Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

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1298 Fermented Unripe Plantain (Musa paradisiacal) Peel Meal as a Replacement for Maize in the Diet of Nile Tilapia (Oreochromis niloticus) Fingerlings

Authors: N. A. Bamidele, S. O. Obasa, I. O. Taiwo, I. Abdulraheem, O. C. Odebiyi, A. A. Adeoye, O. E. Babalola, O. V. Uzamere

Abstract:

A feeding trial was conducted to investigate the effect of fermented unripe plantain peel meal (FUP) on growth performance, nutrients digestibility and economic indices of production of Nile tilapia, Oreochromis niloticus fingerlings. Fingerlings (150) of Nile tilapia (1.70±0.1g) were stocked at 10 per plastic tank. Five iso-nitrogenous diets containing 40% crude protein in which maize meal was replaced by fermented unripe plantain peel meal at 0% (FUP0), 25% (FUP25), 50% (FUP50), 75% (FUP75) and 100% (FUP100) were formulated and prepared. The fingerlings were fed at 5% body weight per day for 56 days. There was no significant difference (p > 0.05) in all the growth parameters among the treatments. Feed conversion ratio of 1.35 in fish fed diet FUP25 was not significantly different (P > 0.05) from 1.42 of fish fed diet FUP0. Apparent protein digestibility of 86.94% in fish fed diet FUP100 was significantly higher (p < 0.05) than 70.37% in fish fed diet FUP0 while apparent carbohydrate of 88.34% in fish fed diet FUP0 was significantly different (p < 0.05) from 70.29% of FUP100. Red blood cell (4.30 ml/mm3) of fish fed diet FUP100 was not significantly different from 4.13 ml/mm3 of fish fed diet FUP50. The highest percentage profit of 88.85% in fish fed diet FUP100 was significantly higher than 66.68% in fish fed diet FUP0 while the profit index of 1.89 in fish fed diet FUP100 was significantly different from 1.67 in fish fed diet FUP0. Therefore, fermented unripe plantain peel meal can completely replace maize in the diet of O. niloticus fingerlings.

Keywords: fermentation, fish diets, plantain peel, tilapia

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1297 Electrochemical and Photoelectrochemical Study of Polybithiophene–MnO2 Composite Films

Authors: H. Zouaoui, D. Abdi, B. Nessark, F. Habelhames, A. Bahloul

Abstract:

Among the conjugated organic polymers, the polythiophenes constitute a particularly important class of conjugated polymers, which has been extensively studied for the relation between the geometrical structure and the optic and electronic properties, while the polythiophene is an intractable material. They are, furthermore, chemically and thermally stable materials, and are very attractive for exploitation of their physical properties. The polythiophenes are extensively studied due to the possibility of synthesizing low band gap materials by using substituted thiophenes as precursors. Low band gap polymers may convert visible light into electricity and some photoelectrochemical cells based on these materials have been prepared. Polythiophenes (PThs) are good candidates for polymer optoelectronic devices such as polymer solar cells (PSCs) polymer light-emitting diodes (PLEDs) field-effect transistors (FETs) electrochromics and biosensors. In this work, MnO2 has been synthesized by hydrothermal method and analyzed by infrared spectroscopy. The polybithiophene+MnO2 composite films were electrochemically prepared by cyclic voltammetry technic on a conductor glass substrate ITO (indium–tin-oxide). The composite films are characterized by cyclic voltammetry, impedance spectroscopy and photoelectrochemical analyses. The results confirmed the presence of manganese dioxide nanoparticles in the polymer layer. An application has been made by using these deposits as an electrode in a photoelectrochemical cell for measuring photocurrent tests. The composite films show a significant photocurrent intensity 80 μA.cm-2.

Keywords: polybithiophene, MnO2, photoelectrochemical cells, composite films

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1296 Investigation of Polymer Solar Cells Degradation Behavior Using High Defect States Influence Over Various Polymer Absorber Layers

Authors: Azzeddine Abdelalim, Fatiha Rogti

Abstract:

The degradation phenomenon in polymer solar cells (PCSs) has not been clearly explained yet. In fact, there are many causes that show up and influence these cells in a variety of ways. Also, there has been a growing concern over this degradation in the photovoltaic community. One of the main variables deciding PSCs photovoltaic output is defect states. In this research, devices modeling is carried out to analyze the multiple effects of degradation by applying high defect states (HDS) on ideal PSCs, mainly poly(3-hexylthiophene) (P3HT) absorber layer. Besides, a comparative study is conducted between P3HT and other PSCs by a simulation program called Solar Cell Capacitance Simulator (SCAPS). The adjustments to the defect parameters in several absorber layers explain the effect of HDS on the total output properties of PSCs. The performance parameters for HDS, quantum efficiency, and energy band were therefore examined. This research attempts to explain the degradation process of PSCs and the causes of their low efficiency. It was found that the defects often affect PSCs performance, but defect states have a little effect on output when the defect level is less than 1014cm-3, which gives similar performance values with P3HT cells when these defects is about 1019cm-3. The high defect states can cause up to 11% relative reduction in conversion efficiency of ideal P3HT. In the center of the band gap, defect states become more noxious. This approach is for one of the degradation processes potential of PSCs especially that use fullerene derivative acceptors.

Keywords: degradation, high defect states, polymer solar cells, SCAPS-1D

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1295 Effects of Cymbopogon citratus, Stapf (CS) or Lemon Grass Ethanol Extract on Antioxidant and Vascular Disorders Parameters in Rat

Authors: Suphaket Saenthaweesuk, Nutiya Somparn, Atcharaporn Thewmore

Abstract:

The present study aims to investigate the effects of Cymbopogon citratus, Stapf (CS) or lemon grass ethanol extract on antioxidant and vascular disorders parameters in rat. The CS ethanol extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with CS at 1,000 mg/kg/day for 30 days. Phytochemical screening of CS extract indicated the presence of tannins, flavonoids and phenolic compounds. The extract contained phenolic compounds 1,400.10 ± 0.47 mg of gallic acid equivalents per gram CS extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 168.77 ± 3.32µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14 µg/mL. In the animals, the protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased. This was consistent with elevation of serum catalase (CAT) and superoxide dismutase (SOD) activities. However, Protein expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule (ICAM-1) and endothelial nitric oxide synthase (eNOS) in heart and aorta were not differenced from normal control. Taken together, the present study provides evidence that CCS water extract exhibits direct antioxidant properties and can induce cytoprotective enzymes in vivo.

Keywords: antioxidant, Cymbopogon citratus Stapf, VCAM-1, γ-glutamylcysteine ligase

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1294 Production of Biotechnological Chondroitin from Recombinant E, Coli K4 Strains on Renewable Substrates

Authors: Donatella Cimini, Sergio D’ambrosio, Saba Sadiq, Chiara Schiraldi

Abstract:

Chondroitin sulfate (CS), as well as modified CS, and unsulfated chondroitin, are largely applied in research today. CS is a linear glycosaminoglycan normally present in cartilage-rich tissues and bones in the form of proteoglycans decorated with sulfate groups in different positions. CS is used as an effective non-pharmacological alternative for the treatment of osteoarthritis, and other potential applications in the biomedical field are being investigated. Some bacteria, such as E. coli K4, produce a polysaccharide that is a precursor of CS (unsulfated chondroitin). This work focused on the construction of integrative E. coli K4 recombinant strains overexpressing genes (kfoA, kfoF, pgm and galU in different combinations) involved in the biosynthesis of the nucleotide sugars necessary for polysaccharide synthesis. Strain growth and polymer production were evaluated using renewable waste materials as substrates in shake flasks and small-scale batch fermentation processes. Results demonstrated the potential to replace pure sugars with cheaper medium components to establish environmentally sustainable and cost-effective production routes for potential industrial development. In fact, although excellent fermentation results have been described so far by employing strains that naturally produce chondroitin-like polysaccharides on semi-defined media, there is still the need to reduce manufacturing costs by providing a cost-effective biotechnological alternative to currently used animal-based extraction procedures.

Keywords: E. coli K4, chondroitin, microbial cell factories, glycosaminoglycans, renewable resources

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1293 The Exposure to Endocrine Disruptors during Pregnancy and Relation to Steroid Hormones

Authors: L. Kolatorova, J. Vitku, K. Adamcova, M. Simkova, M. Hill, A. Parizek, M. Duskova

Abstract:

Endocrine disruptors (EDs) are substances leaching from various industrial products, which are able to interfere with the endocrine system. Their harmful effects on human health are generally well-known, and exposure during fetal development may have lasting effects. Fetal exposure and transplacental transport of bisphenol A (BPA) have been recently studied; however, less is known about alternatives such as bisphenol S (BPS), bisphenol F (BPF) and bisphenol AF (BPAF), which have started to appear in consumer products. The human organism is usually exposed to the mixture of EDs, out of which parabens are otherwise known to transfer placenta. The usage of many cosmetic, pharmaceutical and consumer products during the pregnancy that may contain parabens and bisphenols has led to the need for investigation. The aim of the study was to investigate the transplacental transport of BPA, its alternatives, and parabens, and to study their relation to fetal steroidogenesis. BPA, BPS, BPF, BPAF, methylparaben, ethylparaben, propylparaben, butylparaben, benzylparaben and 15 steroids including estrogens, corticoids, androgens and immunomodulatory ones were determined in 27 maternal (37th week of gestation) and cord plasma samples using liquid chromatography - tandem mass spectrometry methods. The statistical evaluation of the results showed significantly higher levels of BPA (p=0.0455) in cord plasma compared to maternal plasma. The results from multiple regression models investigated that in cord plasma, methylparaben, propylparaben and the sum of all measured parabens were inversely associated with testosterone levels. To our best knowledge, this study is the first attempt to determine the levels of alternative bisphenols in the maternal and cord blood, and also the first study reporting the simultaneous detection of bisphenols, parabens, and steroids in these biological fluids. Our study confirmed the transplacental transport of BPA, with likely accumulation in the fetal compartment. The negative association of cord blood parabens and testosterone levels highlights their possible risks, especially for the development of male fetuses. Acknowledgements: This work was supported by the project MH CR 17-30528 A from the Czech Health Research Council, MH CZ - DRO (Institute of Endocrinology - EÚ, 00023761) and by the MEYS CR (OP RDE, Excellent research - ENDO.CZ).

Keywords: bisphenol, endocrine disruptor, paraben, pregnancy, steroid

Procedia PDF Downloads 180
1292 Binding Studies and Structure Determination of the Recombinantly Produced Type-II 3-Dehydroquinate Dehydratase from Acinetobacter Baumannii

Authors: Naseer Iqbal, Mukesh Kumar, Pradeep Sharma, Satya Prakash Yadav, Punit Kaur, Sujata Sharma, T. P. Singh

Abstract:

Dehydroquinase (3-dehydroquinate dehydratase, DHQD, EC 4.2.1.10) is involved in shikimate pathway and catalyzes the conversion of dehydroquinate to dehydroshikimate. Shikimate pathway is important drug target as this pathway is absent in mammals. DHQD from Acinetobacter baumannii (AbDHQD) was cloned, expressed and purified to homogeneity. The binding studies showed that compounds quinic acid and citrazinic acid bound to AbDHQD at micromolar concentrations. AbDHQD was crystallized using 30% PEG-3350, 50mM tris-HCl, and 1.0M MgSO4 at PH 8.0. Crystals of AbDHQD were stabilized by transferring them into reservoir solution to which 25% glycerol was added for data collection at 100K. The X-ray intensity data were collected to 2.0Å resolution. The crystals belong to monoclinic space group P21 with cell dimensions, a = 82.3, b = 95.3, c = 132.3Å and β = 95.7°. The structure was solved with molecular replacement method and refined to Rcryst/Rfree factors of 0.200/0.232. The structures of 12 crystallographically independent molecules in the asymmetry unit were identical with r.m.s shifts for the C atoms ranging from 0.3 Å to 0.8 Å. They formed a dodecamer with four trimers arranged in a tetrahedral manner. The classical lid adopted an open conformation although a sulfate ion was observed in the substrate binding site. As a result of which, the compounds quinic acid and citrazinic acid did not bind to AbDHQD.

Keywords: acinetobacter Bauman Nii, dehydroquinate dehydratase, dodecamer, open conformation

Procedia PDF Downloads 363
1291 Comparison the Effect of Different Pretreatments on Ethanol Production from Lemon Peel (Citrus × latifolia)

Authors: Zohreh Didar Yaser, Zanganeh Asadabadi

Abstract:

The aim of this work is to open up the structure of lemon peel (Citrus × latifolia) with mild pretreatments. The effects of autoclave, microwave and ultrasonic with or without acid addition were investigated on the amount of glucose, soluble and insoluble lignin, furfural, yeast viability and bioethanol. The finding showed that autoclave- acid impregnated sample, has the highest glucose release from lignocellulose materials (14.61 and 14.95 g/l for solvent exposed and untreated sample, respectively) whereas at control sample glucose content was at its minimal level. Pretreatments cause decrease on soluble and insoluble lignin and the highest decrease cause by autoclave following with microwave and ultrasonic pretreatments (p≤5%). Moderate increase on furfural was seen at pretreated samples than control ones. Also, the most yeast viability and bioethanol content was belong to autoclave samples especially acid- impregnated ones (40.33%). Comparison between solvent treated and untreated samples indicated that significant difference was between two tested groups (p≤1%) in terms of lignin, furfural, cell viability and ethanol content but glucose didn’t show significant difference. It imply that solvent extraction don’t influences on glucose release from lignocellulose material of lemon peel but cause enhancement of yeast viability and bioethanol production.

Keywords: Bioethanol, Lemon peel, Pretreatments, Solvent Extraction

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1290 Catalytic Pyrolysis of Sewage Sludge for Upgrading Bio-Oil Quality Using Sludge-Based Activated Char as an Alternative to HZSM5

Authors: Ali Zaker, Zhi Chen

Abstract:

Due to the concerns about the depletion of fossil fuel sources and the deteriorating environment, the attempt to investigate the production of renewable energy will play a crucial role as a potential to alleviate the dependency on mineral fuels. In this respect, biofuels are measured as a vital nominee for national energy security and energy sustainability. Sewage sludge (SS), as an alternative source of renewable energy with a complex composition, is a major waste generated during wastewater treatment. Stricter legislation is continuously refining the requirements for the level of removal of various pollutants in treated water, causing continuous growth of sludge production, which has become a global challenge. In general, there are two main procedures for dealing with SS: incineration and landfill. However, there are a variety of limitations in these options (e.g., production of greenhouse gases and restrictive environmental regulations) in regard to negative social and economic impacts. Pyrolysis is a feasible and cost-effective technology that can simultaneously tackle boundaries concerning the current disposal routes while retrieving bioenergy. Pyrolysis of SS has drawn vigorous interest in research due to the ability of high mass yield of pyrolytic liquid production. Nonetheless, the presence of high molecular weight hydrocarbons and oxygenated- and nitrogenated compounds poses a considerable challenge. In this context, catalytic pyrolysis is another attainable route in order to upgrade the bio-oil quality. Among different catalysts (i.e., zeolites) studied for sewage sludge pyrolysis, activated chars are eco-friendly and low-cost alternatives. The beneficial features comprise comparatively large surface area, long-term stability, and enriched surface functional groups. In light of these premises, this research attempts to investigate the catalytic pyrolysis of sewage sludge with a high-performance sludge-based activated char in contrast to HZSM5 from a theoretical and experimental point of view.

Keywords: catalytic pyrolysis, sewage sludge, char, HZSM5, bio-oil.

Procedia PDF Downloads 52
1289 Competition Between the Effects of Pesticides and Immune-activation on the Expression of Toll Pathway Genes

Authors: Dani Sukkar, Ali Kanso, Philippe Laval-Gilly, Jairo Falla-Angel

Abstract:

The honeybees' immune system is challenged by different risk factors that induce various responses. However, complex scenarios where bees are exposed to different pesticides simultaneously with immune activation are not well evaluated. The Toll pathway is one of the main signaling pathways studied in invertebrate immune responses, and it is a good indicator of the effect of such complex interactions in addition to key signaling elements of other pathways like Relish of the immune deficiency (IMD) pathway or Eater, the phagocytosis receptor or vitellogenin levels. Honeybee hemocytes extracted from 5th instar larvae were exposed to imidacloprid and/or amitraz with or without the presence of the zymosan a as an immune activator. The gene expression of multiple immune related genes were studied, including spaetzle, Toll, myD88, relish, eater and vitellogenin, by real-time polymerase chain reaction after RNA extraction. The results demonstrated that the Toll pathway is mainly affected by the pesticides; imidacloprid and amitraz, especially by their different combinations. Furthermore, immune activation by zymosan A, a fungal cell-wall component, acts to mitigate to some extent the effect of pesticides on the different levels of the Toll pathway. In addition, imidacloprid, amitraz, and zymosan A have complex and context-specific interactions depending on the levels of immune activation and the pathway evaluated affecting immune-gene expression differently.

Keywords: toll pathway, immune modulation, β-glucan, imidacloprid, amitraz, honeybees, immune genes

Procedia PDF Downloads 91
1288 Thiazolo[5,4-D]Thiazole-Core Organic Chromophore with Furan Spacer for Organic Solar Cells

Authors: M. Nazim, S. Ameen, H. K. Seo, H. S. Shin

Abstract:

Energy is the basis of life and strong attention has been growing for the cost-effective energy production. Recently, solution-processed small molecule organic solar cells (SMOSCs) have grown much attention due to the wages such as well-defined molecular structures, definite molecular weight, easy synthesis and easy purification techniques. In particular, the size of donor (D) and acceptor (A) unit is a crucial factor for the exciton-diffusion towards D-A interface and then charge-separation for the effective charge-transport to the electrodes. Furan-bridged materials are more electron-rich, high fluorescence, with better molecular-packing, and greater rigidity and greater solubility than their thiophene-counterparts In this work, a furan-bridged thiazolo[5,4-d]thiazole based organic small molecule (RFTzR) was formulated and applied for BHJ organic solar cells (OSCs). The introduction of furan spacer with two terminal alkyl units improved its absorption and solubility in the common organic solvents, significantly. RFTzR exhibited a HOMO and LUMO energy levels of -5.36 eV and -3.14 eV, respectively. The fabricated solar cell devices of RFTzR (donor) with PC60BM (acceptor) as photoactive materials showed high performance of 2.72% (RFTzR:PC60BM, 2:1, w/w) ratio with open-circuit voltage of 0.756 V and high photocurrent density of 10.13 mA/cm².

Keywords: chromophore, organic solar cells, photoactive materials, small molecule

Procedia PDF Downloads 164
1287 The Effects of Ultrasound on the Extraction of Ficus deltoidea Leaves

Authors: Nur Aimi Syairah Mohd Abdul Alim, Azilah Ajit, A. Z. Sulaiman

Abstract:

The present study aimed to investigate the effects of ultrasound-assisted extraction (UAE) on the extraction of Vitexin and Iso-Vitexin from Ficus deltoidea plants. In recent years, ultrasound technology has been found to be a potential herbal extraction technique. The passage of ultrasound energy in a liquid medium generates mechanical agitation and other physical effects due to acoustic cavitation. The main goal is to optimised ultrasonic-assisted extraction condition providing the highest extraction yield with the most desirable antioxidant activity and stability. Thus, a series of experiments has been developed to investigate the effect of ultrasound energy on the vegetal material and the implemented parameters by using HPLC-photodiode array detection. The influences of several experimental parameters on the ultrasonic extraction of Ficus deltoidea leaves were investigated: extraction time (1-8 h), solvent-to-water ratio (1:10 to 1:50), temperature (50–100 °C), duty cycle (10–continuous sonication) and intensity. The extracts at the optimized condition were compared with those obtained by conventional boiling extraction, in terms of bioactive constituents yield and chemical composition. The compounds of interest identified in the extracts were Vitexin and Isovitexin, which possess anti-diabetic, anti-oxidant and anti-cancer properties. Results showed that the main variables affecting the extraction process were temperature and time. Though in less extent, solvent-to-water ratio, duty cycle and intensity are also demonstrated to be important parameters. The experimental values under optimal conditions were in good consistent with the predicted values, which suggested that ultrasonic-assisted extraction (UAE) is more efficient process as compared to conventional boiling extraction. It recommended that ultrasound extraction of Ficus deltoidea plants are feasible to replace the traditional time-consuming and low efficiency preparation procedure in the future modernized and commercialized manufacture of this highly valuable herbal medicine.

Keywords: Ficus, ultrasounds, vitexin, isovitexin

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1286 Survival Outcomes Related to Treatment Modalities in Patients with Oropharyngeal Squamous Cell Carcinoma

Authors: Danni Cheng

Abstract:

Purpose:Surgicallyinclusive treatment(SIT)isthemajor treatment fororopharyngealsquamouscellcarcinoma (OPSCC) in Eastern countries, while nonsurgical treatments(NSTs) are the priority treatment in Western countries. The preferred treatmentsforOPSCC patients remaindebated. Methods:Atotalof 153 consecutive OPSCC casesdiagnosed between 2009 and 2019inWCH, and 15,400 OPSCC cases from SEER database (2000-2017) were obtained. Clinical characteristics, treatments, and survival outcomes were retrospectively collected. We conductedKaplan-Meier curves univariate and multivariate analysis to compare the prognosis of OPSCC patients in WCH, SEER Asian, and SEER all ethnic population by different treatment modalities,HPVstatus, ages, and TNM stages. Results: The 5-year overall survival rate was 59% in WCH, 64% in the SEER all ethnic and 67% in SEER Asian group. In both univariate and multivariate analysis, SIT was observed as a consistent benefit factor for OPSCC patients in all three populations when classified by genders, tumor stages, and HPV status. Patients who underwent SIT had significantly better survival outcomes than those who received NSTsin WCH, SEER Asian, and SEER all ethnic groups. HPV positive status was the beneficial factor of OPSCC patients in all three groups. Besides, male patients had worse survival outcomes in both WCH and SEER Asian group, whereas male patients had better outcomes in the SEER all ethnic group. Conclusion: In contrast to nowadaysNSTs are the first-line therapiesfor OPSCC, our ten-year real-world data and SEER data indicated that OPSCC patients who underwent SIT had better prognosis than NSTs.

Keywords: OPSCC, survival outcome, SEER, treatment modalities

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1285 Comparative Analysis of Single vs. Multiple gRNA on NGN3 Expression Using a Controllable dCas9-VP192 Activator (CRISPRa)

Authors: Nicholas Abdilmasih, Habib Rezanejad

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This study investigates the gene expression induction efficiency of single versus multiple guide RNAs (gRNAs) targeting the NGN3 gene using the CRISPR activation system in HEK293 cells. Our study aimed to contribute to optimizing the use of gRNAs in gene therapy applications, particularly in treating diseases like diabetes, where precise gene regulation is essential. The experimental design involves culturing HEK293 cells, and once they reach approximately 70-80% confluence, cells were transfected with specific gRNAs targeting the NGN3 gene promoter. Specific gRNAs targeting the NGN3 promoter that was previously designed, incorporated into plasmid clone cassettes and introduced into HEK293 cells through co-transfection using pCAG-DDdCas9-VP192-EGFP transactivator. Post-transfection, cell viability, and fluorescence were monitored to assess transfection efficiency. RNA was extracted, converted to cDNA, and analyzed via qPCR to measure NGN3 expression levels. Results indicated that specific combinations of fewer gRNAs led to higher NGN3 activation compared to multiple gRNAs, challenging the assumption that more gRNAs result in synergistic gene activation. These findings suggest that optimized gRNA combinations can enhance gene therapy efficiency, potentially leading to more effective treatments for conditions like diabetes.

Keywords: CRISPR activation, Diabetes mellitus, gene therapy, guide RNA, Neurogenin3

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