Search results for: buffer

Authors: Nasser A. Al-Shabib

Abstract:

Food allergens are most common non-native form when exposed to the immune system. Most food proteins undergo various treatments (e.g. thermal or proteolytic processing) during food manufacturing. Such treatments have the potential to impact the chemical structure of food allergens so as to convert them to more denatured or unfolded forms. The conformational changes in the proteins may affect the allergenicity of treated-allergens. However, most allergenic proteins possess high resistance against thermal modification or digestive enzymes. In the present study, ovomucoid (a major allergenic protein of egg white) was heated in phosphate-buffered saline (pH 7.4) at different temperatures, aqueous solutions and on different surfaces for various times. The results indicated that different antibody-based methods had different sensitivities in detecting the heated ovomucoid. When using one particular immunoassay‚ the immunoreactivity of ovomucoid increased rapidly after heating in water whereas immunoreactivity declined after heating in alkaline buffer (pH 10). Ovomucoid appeared more immunoreactive when dissolved in PBS (pH 7.4) and heated on a stainless steel surface. To the best of our knowledge‚ this is the first time that antibody-based methods have been applied for the detection of ovomucoid adsorbed onto different surfaces under various conditions. The results obtained suggest that use of antibodies to detect ovomucoid after food processing may be problematic. False assurance will be given with the use of inappropriate‚ non-validated immunoassays such as those available commercially as ‘Swab’ tests. A greater understanding of antibody-protein interaction after processing of a protein is required.

Keywords: ovomucoid, thermal treatment, solutions, surfaces

Procedia PDF Downloads 448

Authors: Doriane Delafosse, Dominique Fontvieille

Abstract:

Background: Wastewater treatment plants (WWTPs) generally display significant efficacy in virus retention yet, are sometimes highly variable, partly in relation to large fluctuating loads at the head of the plant and partly because of episodic dysfunctions in some treatment processes. The problem is especially sensitive when human enteric viruses, such as human Noroviruses Genogroup I or Adenoviruses, are in concern: their release downstream WWTP, in environments often interconnected to recreational areas, may be very harmful to human communities even at low concentrations. It points out the importance of WWTP permanent monitoring from which their internal treatment processes could be adjusted. One way to adjust primary treatments is to add coagulants and flocculants to sewage ahead settling tanks to improve decantation. In this work, sludge produced by three coagulants (two organics, one mineral), four flocculants (three cationic, one anionic), and their combinations were studied for their efficacy in human enteric virus retention. Sewage samples were coming from a WWTP in the vicinity of the laboratory. All experiments were performed three times and in triplicates in laboratory pilots, using Murine Norovirus (MNV-1), a surrogate of human Norovirus, as an internal control (spiking). Viruses were quantified by (RT-)qPCR after nucleic acid extraction from both treated water and sediment. Results: Low values of sludge virus retention (from 4 to 8% of the initial sewage concentration) were observed with each cationic organic flocculant added to wastewater and no coagulant. The largest part of the virus load was detected in the treated water (48 to 90%). However, it was not counterbalancing the amount of the introduced virus (MNV-1). The results pertained to two types of cationic flocculants, branched and linear, and in the last case, to two percentages of cations. Results were quite similar to the association of a linear cationic organic coagulant and an anionic flocculant, though suggesting that differences between water and sludges would sometimes be related to virus size or virus origins (autochthonous/allochthonous). FeCl₃, as a mineral coagulant associated with an anionic flocculant, significantly increased both auto- and allochthonous virus retention in the sediments (15 to 34%). Accordingly, virus load in treated water was lower (14 to 48%) but with a total that still does not reach the amount of the introduced virus (MNV-1). It also appeared that the virus retrieval in a bare 0.1M NaCl suspension varied rather strongly according to the FeCl₃ concentration, suggesting an inhibiting effect on the molecular analysis used to detect the virus. Finally, no viruses were detected in both phases (sediment and water) with the combination branched cationic coagulant-linear anionic flocculant, which was later demonstrated as an effect, here also, of polymers on the virus detection-molecular analysis. Conclusions: The combination of FeCl₃-anionic flocculant gave its highest performance to the decantation-based virus removal process. However, large unbalanced values in spiking experiments were observed, suggesting that polymers cast additional obstacles to both elution buffer and lysis buffer on their way to reach the virus. The situation was probably even worse with autochthonous viruses already embedded into sewage's particulate matter. Polymers and FeCl₃ also appeared to interfere in some steps of molecular analyses. More attention should be paid to such impediments wherever chemical additives are considered to be used to enhance WWTP processes. Acknowledgments: This research was supported by the ABIOLAB laboratory (Montbonnot Saint-Martin, France) and by the ASPOSAN association. Field experiments were possible thanks to the Grand Chambéry WWTP authorities (Chambéry, France).

Keywords: flocculants-coagulants, polymers, enteric viruses, wastewater sedimentation treatment plant

Procedia PDF Downloads 125

Authors: Reyhaneh Saeedi

Abstract:

Iran is one of the regions which are most prone for earthquakes annually having a large number of financial and mortality and financial losses. Every year around the world, a large number of people lose their home and life due to natural disasters such as earthquakes. It is necessary to provide and specify some suitable places for settling the homeless people before the occurrence of the earthquake, one of the most important factors in crisis planning and management. Some of the natural disasters can be Modeling and shown by Geospatial Information System (GIS). By using GIS, it would be possible to manage the spatial data and reach several goals by making use of the analyses existing in it. GIS has a determining role in disaster management because it can determine the best places for temporary resettling after such a disaster. In this research QuantumGIS software is used that It is an OpenSource software so that easy to access codes and It is also free. In this system, AHP method is used as decision model and to locate the best places for temporary resettling, is done based on the related organizations criteria with their weights and buffers. Also in this research are made the buffer layers of criteria and change them to the raster layers. Later on, the raster layers are multiplied on desired weights then, the results are added together. Eventually, there are suitable places for resettling of victims by desired criteria by different colors with their optimum rate in QuantumGIS platform.

Keywords: disaster management, temporary resettlement, earthquake, QuantumGIS

Procedia PDF Downloads 398

Authors: Eniola Simbiat Ibude

Abstract:

Work politics could be referred to as office politics or organizational politics. Work place politics take different form, direction, and dimensions. Studies of these features of organizational politics have been conducted in the private sector and much has been left to be studied on the other side of the fence, namely in larger bureaucracies and in public sector system. This is the gap the study tried to fill. This study also focuses on the negative effects that perceptions of politics seem to have on job attitudes (i.e., job satisfaction, organizational commitment) and on affective performance. This was with a view to understanding the relevance of its effects on job performance. The descriptive survey research design of the ex-post facto type was adopted for this study since the variables being studied had already occurred and were, therefore, not manipulated. Data were analyzed using the descriptive and inferential statistics of frequency counts, simple percentages, ANOVA, and multiple regression. Findings show that the joint and relative effect of organizational politics on workers performance, planning, coordination and supervision of work (B 0.71), delaying information for carrying out work (B 0.67), criticizing and wasting time for work done (B 0.56) has contributed to workers performance. The effect could be seen as negative on workers performance. Conclusively, every employee will not react to organizational politics the same way. The 'social arsenal' or the 'social skills' of the individual are a good buffer against the potential aftermaths of organizational politics. Also, from this study, it could be concluded that the perceptions of politics have a more complex relationship with job performance, a relationship that may be different for various types of employees.

Keywords: bureaucracies, dimension, politics, trend

Procedia PDF Downloads 242

Authors: Giovanni Meloni

Abstract:

Recent results are presented from experiments carried out at the Advanced Light Source (ALS) at the Chemical Dynamics Beamline of Lawrence Berkeley National Laboratory using multiplexed synchrotron photoionization mass spectrometry. The reaction mixture and a buffer gas (He) are introduced through individually calibrated mass flow controllers into a quartz slow flow reactor held at constant pressure and temperature. The gaseous mixture effuses through a 650 μm pinhole into a 1.5 mm skimmer, forming a molecular beam that enters a differentially pumped ionizing chamber. The molecular beam is orthogonally intersected by a tunable synchrotron radiation produced by the ALS in the 8-11 eV energy range. Resultant ions are accelerated, collimated, and focused into an orthogonal time-of-flight mass spectrometer. Reaction species are identified by their mass-to-charge ratios and photoionization (PI) spectra. Comparison of experimental PI spectra with literature and/or simulated curves is routinely done to assure the identity of a given species. With the aid of electronic structure calculations, potential energy surface scans are performed, and Franck-Condon spectral simulations are obtained. Examples of these experiments are discussed, ranging from new intermediates characterization to reaction mechanisms elucidation and biofuels oxidation pathways identification.

Keywords: mass spectrometry, reaction intermediates, synchrotron photoionization, oxidation reactions

Procedia PDF Downloads 73

Authors: H. Abu-Ali, A. Nabok, T. Smith

Abstract:

Aptamers are single-strand of DNA or RNA nucleotides sequence which is designed in vitro using selection process known as SELEX (systematic evolution of ligands by exponential enrichment) were developed for the selective detection of many toxic materials. In this work, we have developed an electrochemical biosensor for highly selective and sensitive detection of Hg2+ and Pb2+ using a specific aptamer probe (SAP) labelled with ferrocene (or methylene blue) in (5′) end and the thiol group at its (3′) termini, respectively. The SAP has a specific coil structure that matching with G-G for Pb2+ and T-T for Hg2+ interaction binding nucleotides ions, respectively. Aptamers were immobilized onto surface of screen-printed gold electrodes via SH groups; then the cyclic voltammograms were recorded in binding buffer with the addition of the above metal salts in different concentrations. The resulted values of anode current increase upon binding heavy metal ions to aptamers and analyte due to the presence of electrochemically active probe, i.e. ferrocene or methylene blue group. The correlation between the anodic current values and the concentrations of Hg2+ and Pb2+ ions has been established in this work. To the best of our knowledge, this is the first example of using a specific DNA aptamers for electrochemical detection of heavy metals. Each increase in concentration of 0.1 μM results in an increase in the anode current value by simple DC electrochemical test i.e (Cyclic Voltammetry), thus providing an easy way of determining Hg2+ and Pb2+concentration.

Keywords: aptamer, based, biosensor, DNA, electrochemical, highly, specific

Procedia PDF Downloads 161

Authors: Azizza Mala, Babo Fadlalla, Elnour Mohamed, Siran Wang, Junfeng Li, Tao Shao

Abstract:

Sweet sorghum is considered one of the best plants for silage production and is now a more important feed crop in many countries worldwide. It is simple to ensile because of its high water-soluble carbohydrates (WSC) concentration and low buffer capacity. This study investigated the effect of adding Pediococcus acidilactici AZZ5 and Lactobacillus plantarum AZZ4 isolated from elephant grass on the fermentation quality of sweet sorghum silage. One commercial bacteria Lactobacillus Plantarum, Ecosyl MTD/1(C.B.)), and two strains were used as additives Pediococcus acidilactici (AZZ5), Lactobacillus plantarum subsp. Plantarum (AZZ4) at 6 log colony forming units (cfu)/g of fresh sweet sorghum grass in laboratory silos (1000g). After 15, 30, and 60 days, the silos for each treatment were opened. All of the isolated strains enhanced the silage quality of sweet sorghum silage compared to the control, as evidenced by significantly (P < 0.05) lower ammonia nitrogen (NH3-N) content and undesirable microbial counts, as well as greater lactic acid (L.A.) contents and lactic acid/acetic acid (LA/AA) ratios. In addition, AZZ4 performed better than all other inoculants during ensiling, as evidenced by a significant (P < 0.05) reduction in pH and ammonia-N contents and a significant increase in lactic acid contents.

Keywords: fermentation, lactobacillus plantarum, lactic acid bacteria, pediococcus acidilactic, sweet sorghum

Procedia PDF Downloads 92

Authors: Hati̇ce Bi̇rtane, Asli Beyler Çi̇ği̇l, Memet Vezi̇r Kahraman

Abstract:

Polyimide (PI) is one of the most important super-engineering materials because of its mechanical properties and its thermal stability. Electronic industry is the typical extensive applications of polyimides including interlayer insulation films, buffer coating, films, alpha-ray shielding films, and alignment films for liquid crystal displays. The mechanical and thermal properties of polymers are generally improved by the addition of inorganic additives. The challenges in this area of high-performance organic/inorganic hybrid materials are to obtain significant improvements in the interfacial adhesion between the polymer matrix and the reinforcing material since the organic matrix is relatively incompatible with the inorganic phase. In this study, modified nanodiamond was prepared from the reaction of nanodiamond and (3-Mercaptopropyl)trimethoxysilane. Poly(amic acid) was prepared from the reaction of 3,3',4,4'-Benzophenonetetracarboxylic dianhydride (BTDA) and 4,4'-Oxydianiline (ODA). Polyimide/modified nanodiamond hybrids were prepared by blending of poly(amic acid) and organically modified nanodiamond. The morphology of the Polyimide/ modified nanodiamond hybrids was characterized by scanning electron microscopy (SEM). Chemical structure of polyimide and Polyimide/modified nanodiamond hybrids was characterized by FTIR. FTIR results showed that the Polyimide/modified nanodiamond hybrids were successfully prepared. A thermal property of the Polyimide/modified nanodiamond hybrids was characterized by thermogravimetric analysis (TGA).

Keywords: hybrid materials, nanodiamond, polyimide, polymer

Procedia PDF Downloads 243

Authors: Mahnoosh Aliahmadi, Akbar Esmaeili

Abstract:

This study synthesized new modified imaging nanocapsules (NCs) of gallium@deferoxamine/folic acid/chitosan/polyaniline/polyvinyl alcohol (Ga@DFA/FA/CS/PANI/PVA) containing Morus nigra extract by selenium nanoparticles prepared from Lactobacillus acidophilus. Se nanoparticles were then deposited on (Ga@DFA/FA/CS/PANI/PVA) using the impregnation method. The modified contrast agents were mixed with M. nigra extract, and their antibacterial activities were investigated by applying them to L929 cell lines. The influence of variable factors including surfactant, solvent, aqueous phase, pH, buffer, minimum Inhibitory concentration (MIC), minimum bactericidal concentration (MBC), cytotoxicity on cancer cells, antibiotic, antibiogram, release and loading, stirring effect, the concentration of nanoparticle, olive oil, and thermotical methods was investigated. The structure and morphology of the synthesized contrast agents were characterized by zeta potential sizer analysis (ZPS), X-Ray diffraction (XRD), Fourier-transform infrared (FT-IR), and energy-dispersive X-ray (EDX), ultraviolet-visible (UV-Vis) spectra, and scanning electron microscope (SEM). The experimental section was conducted and monitored by response surface methods (RSM) and MTT conversion assay. Antibiogram testing of NCs on Pseudomonas aeruginosa bacteria was successful, and the MIC=2 factor was obtained with a less harmful effect.

Keywords: imaging contrast agent, nanoparticles, response surface method, Lactobacillus acidophilus, selenium

Procedia PDF Downloads 80

Authors: S. Jain, R. Savalia, V. Saini

Abstract:

A simple, accurate, precise, sensitive and specific RP-HPLC method was developed and validated for simultaneous estimation of Metoprolol Succinate and Olmesartan Medoxomil in bulk and tablet dosage form. The RP-HPLC method has shown adequate separation for Metoprolol Succinate and Olmesartan Medoxomil from its degradation products. The separation was achieved on a Phenomenex luna ODS C18 (250mm X 4.6mm i.d., 5μm particle size) with an isocratic mixture of acetonitrile: 50mM phosphate buffer pH 4.0 adjusted with glacial acetic acid in the ratio of 55:45 v/v. The mobile phase at a flow rate of 1.0ml/min, Injection volume 20μl and wavelength of detection was kept at 225nm. The retention time for Metoprolol Succinate and Olmesartan Medoxomil was 2.451±0.1min and 6.167±0.1min, respectively. The linearity of the proposed method was investigated in the range of 5-50μg/ml and 2-20μg/ml for Metoprolol Succinate and Olmesartan Medoxomil, respectively. Correlation coefficient was 0.999 and 0.9996 for Metoprolol Succinate and Olmesartan Medoxomil, respectively. The limit of detection was 0.2847μg/ml and 0.1251μg/ml for Metoprolol Succinate and Olmesartan Medoxomil, respectively and the limit of quantification was 0.8630μg/ml and 0.3793μg/ml for Metoprolol and Olmesartan, respectively. Proposed methods were validated as per ICH guidelines for linearity, accuracy, precision, specificity and robustness for estimation of Metoprolol Succinate and Olmesartan Medoxomil in commercially available tablet dosage form and results were found to be satisfactory. Thus the developed and validated stability indicating method can be used successfully for marketed formulations.

Keywords: metoprolol succinate, olmesartan medoxomil, RP-HPLC method, validation, ICH

Procedia PDF Downloads 315

Authors: Shahana Sharmin

Abstract:

In the present study, the effect of cellulose polymers Ethyl Cellulose and Hydroxy Propyl Methyl Cellulose was evaluated on the release profile of drug from sustained release pellet. Diltiazem Hydrochloride, an antihypertensive, cardio-protective agent and slow channel blocker were used as a model drug to evaluate its release characteristics from different pellets formulations. Diltiazem Hydrochloride sustained release pellets were prepared by drug loading (drug binder suspension) on neutral pellets followed by different percentages of spraying, i.e. 2%,4%, 6%, 8% and 10% coating suspension using ethyl cellulose and hydroxy-propyl methyl cellulose polymer in a fixed 85:15 ratios respectively. The in vitro dissolution studies of Diltiazem Hydrochloride from these sustained release pellets were carried out in pH 7.2 phosphate buffer for 1, 2, 3, 4, 5, 6, 7, and 8 hrs using USP-I method. Statistically, significant differences were found among the drug release profile from different formulations. Polymer content with the highest concentration of Ethyl cellulose on the pellets shows the highest release retarding rate of the drug. The retarding capacity decreases with the decreased concentration of ethyl cellulose. The release mechanism was explored and explained with zero order, first order, Higuchi and Korsmeyer’s equations. Finally, the study showed that the profile and kinetics of drug release were functions of polymer type, polymer concentration & the physico-chemical properties of the drug.

Keywords: diltiazem hydrochloride, ethyl cellulose, hydroxy propyl methyl cellulose, release kinetics, sustained release pellets

Procedia PDF Downloads 414

Authors: Rizky Kusuma Cahyani

Abstract:

Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

Procedia PDF Downloads 151

Authors: Alireza Javed, Keivan Jamshidi

Abstract:

Reactive amyloidosis is a condition that complicates a long list of chronic inflammation, chronic infectious, malignant, and hereditary disorders. In the present study the potential effects of two amyloidogenic substances: ie. AgNO3 and Freund's Adjuvant (AF) with that of vitamin free casein, on spatio-temporal pattern of experimental amyloidosis in mice, were compared. For this purpose, a total of 40 male Swees mice, obtained from Pasteur Institute Tehran, after being weighted were randomly divided into 4 groups including 2 treatments, 1 control (vitamin free casein) and 1 positive control (normal saline). At the end of 3rd, 5th and 7th weeks of experiment 3 mice were randomly selected and euthnised. Spleen sample of each animal obtained and preserved in 10% neutral buffer formalin. Sample were then processed through different stages of dehydration, clearing and impregnation and finally embedded in paraffin blocks. Sections of 5µm thickness then cut and stained by alkaline Congo red techniques. Spleen weights and the data obtained from the microscopic quantitative analysis did show no significant differences between groups A and B, A and C, and B and C. However, significant differences were observed between groups A and D, B and D, and C and D respectively. It is concluded that two compounds ie; AgNO3 and Freund's Adjuvant have the same potential, as does vitamin free casein have, in spatio – temporal pattern of experimental amyloidosis in mice.

Keywords: amyloidosis, mice, AgNO3, Freund's Adjuvant

Procedia PDF Downloads 371

Authors: Imran Ullah Khana, Mohd Hafiz Dzarfan Othmanb, Haslenda Hashima, Takeshi Matsuurad, A. F. Ismailb, M. Rezaei-DashtArzhandib, I. Wan Azelee

Abstract:

Biogas upgrading is a widely studied and discussed topic, and its utilization as a natural gas substitute has gained significant attention in recent years. The production of biomethane provides a versatile application in both heat and power generation and as a vehicular fuel. This paper systematically reviews the state of the art of biogas upgrading technologies with upgrading efficiency, methane (CH4) loss, environmental effect, development and commercialization, and challenges in terms of energy consumption and economic assessment. The market situation for biogas upgrading has changed rapidly in recent years, giving membrane separation a significant market share with traditional biogas upgrading technologies. In addition, the potential utilization of biogas, efficient conversion into bio-compressed natural gas (bio-CNG), and storage systems are investigated in depth. Two storing systems for bio-CNG at filling stations, namely buffer and cascade storage systems are used. The best storage system should be selected on the basis of the advantages of both systems. Also, the fuel economy and mass emissions for bio-CNG and CNG-filled vehicles are studied. There is the same fuel economy and less carbon dioxide (CO2) emission for bio-CNG. Based on the results of comparisons between the technical features of upgrading technologies, various specific requirements for biogas utilization and the relevant investment, and operating and maintenance costs, future recommendations are made for biogas upgrading.

Keywords: biogas upgrading, cost, utilization, bio-CNG, storage, energy

Procedia PDF Downloads 51

Authors: D. Kaczorek

Abstract:

Internal insulation of external walls is often problematic due to increased moisture content in the wall and interstitial or surface condensation risk. In this paper, the hygrothermal performance of prefabricated, concrete, large panel, external wall typical for WK70 system, commonly used in Poland in the 70’s, with inside, additional insulation was investigated. Thermal insulation board made out of hygroscopic, natural materials with moisture buffer capacity and extruded polystyrene (EPS) board was used as interior insulation. Experience with this natural insulation is rare in Poland. The analysis was performed using WUFI software. First of all, the impact of various standard boundary conditions on the behavior of the different wall assemblies was tested. The comparison of results showed that the moisture class according to the EN ISO 13788 leads to too high values of total moisture content in the wall since the boundary condition according to the EN 15026 should be usually applied. Then, hygrothermal 1D-simulations were conducted by WUFI Pro for analysis of internally added insulation, and the weak point like the joint of the wall with the concrete ceiling was verified using 2D simulations. Results showed that, in the Warsaw climate and the indoor conditions adopted in accordance with EN 15026, in the tested wall assemblies, regardless of the type of interior insulation, there would not be any problems with moisture - inside the structure and on the interior surface.

Keywords: concrete large panel wall, hygrothermal simulation, internal insulation, moisture related issues

Procedia PDF Downloads 166

Authors: Rowan Moore, Kai Scheffler, Eugen Damoc, Jennifer Sutton, Aaron Bailey, Stephane Houel, Simon Cubbon, Jonathan Josephs

Abstract:

Purpose: MS analysis of monoclonal antibodies (mAbs) at the protein and peptide levels is critical during development and production of biopharmaceuticals. The compositions of current generation therapeutic proteins are often complex due to various modifications which may affect efficacy. Intact proteins analyzed by MS are detected in higher charge states that also provide more complexity in mass spectra. Protein analysis in native or native-like conditions with zero or minimal organic solvent and neutral or weakly acidic pH decreases charge state value resulting in mAb detection at higher m/z ranges with more spatial resolution. Methods: Three commercially available mAbs were used for all experiments. Intact proteins were desalted online using size exclusion chromatography (SEC) or reversed phase chromatography coupled on-line with a mass spectrometer. For streamlined use of the LC- MS platform we used a single SEC column and alternately selected specific mobile phases to perform separations in either denaturing or native-like conditions: buffer A (20 % ACN, 0.1 % FA) with Buffer B (100 mM ammonium acetate). For peptide analysis mAbs were proteolytically digested with and without prior reduction and alkylation. The mass spectrometer used for all experiments was a commercially available Thermo Scientific™ hybrid Quadrupole-Orbitrap™ mass spectrometer, equipped with the new BioPharma option which includes a new High Mass Range (HMR) mode that allows for improved high mass transmission and mass detection up to 8000 m/z. Results: We have analyzed the profiles of three mAbs under reducing and native conditions by direct infusion with offline desalting and with on-line desalting via size exclusion and reversed phase type columns. The presence of high salt under denaturing conditions was found to influence the observed charge state envelope and impact mass accuracy after spectral deconvolution. The significantly lower charge states observed under native conditions improves the spatial resolution of protein signals and has significant benefits for the analysis of antibody mixtures, e.g. lysine variants, degradants or sequence variants. This type of analysis requires the detection of masses beyond the standard mass range ranging up to 6000 m/z requiring the extended capabilities available in the new HMR mode. We have compared each antibody sample that was analyzed individually with mixtures in various relative concentrations. For this type of analysis, we observed that apparent native structures persist and ESI is benefited by the addition of low amounts of acetonitrile and formic acid in combination with the ammonium acetate-buffered mobile phase. For analyses on the peptide level we analyzed reduced/alkylated, and non-reduced proteolytic digests of the individual antibodies separated via reversed phase chromatography aiming to retrieve as much information as possible regarding sequence coverage, disulfide bridges, post-translational modifications such as various glycans, sequence variants, and their relative quantification. All data acquired were submitted to a single software package for analysis aiming to obtain a complete picture of the molecules analyzed. Here we demonstrate the capabilities of the mass spectrometer to fully characterize homogeneous and heterogeneous therapeutic proteins on one single platform. Conclusion: Full characterization of heterogeneous intact protein mixtures by improved mass separation on a quadrupole-Orbitrap™ mass spectrometer with extended capabilities has been demonstrated.

Keywords: disulfide bond analysis, intact analysis, native analysis, mass spectrometry, monoclonal antibodies, peptide mapping, post-translational modifications, sequence variants, size exclusion chromatography, therapeutic protein analysis, UHPLC

Procedia PDF Downloads 362

Authors: Thuy Hang Le

Abstract:

Traditional agroforestry systems are land-use practices still widespread in tropical and subtropical countries, while in Europe have significantly decreased due to land-use intensification, land abandonment, and urbanization. Nevertheless, scientific evidence reveals that traditional agroforestry systems significantly support biodiversity and ecosystem services and may positively contribute to socioeconomic rural regional development. We worked out a review that follows the PRISMA approach and compiled comprehensive information on traditional agroforestry systems in Europe. Based on the differentiation of different land-use systems, also considering the agricultural as well as forestry components, we compiled information regarding current distribution, management (agrodiversity), biodiversity and agrobiodiversity, ecosystem and landscape services, threats, and restoration initiatives. From a total of 3,304 studies that dealt with agroforestry systems in Europe, both “modern” (e.g., buffer strip) and “traditional” (e.g., meadow orchards), we filtered out 158 studies from 35 European countries which represent the basis for in-depth investigation. We found, for example, that the traditional pastoral agroforestry system in the Mediterranean region, the so-called Dehesa, can harbor up to 300 plant species as well as 238 bird species, of which 134 are breeding birds. With regard to carbon storage, the traditional orchard agroforestry system in Germany stocks ranged between 6.5 and 9.8 Mg C ha−1, showing significantly higher values compared to an intensively used grassland with around 3.4 to 6.7 Mg C ha−1. With the remarkably high benefit for biodiversity and ecosystem services provided, the important role and multifunctionality of traditional agroforestry systems in Europe should be acknowledged and promoted.

Keywords: biodiversity, ecosystem services, landscape services, traditional agroforestry systems

Procedia PDF Downloads 74

Authors: Bert H. Jacobson, Taylor Monaghan, John Sellers

Abstract:

Intense resistance-type activity generally elicits delayed onset muscle soreness (DOMS) in individuals unaccustomed to such action. DOMS is a combination of contractile tissue microtrauma, osmotic pressure changes, alteration calcium regulation, and inflammation. Elevated muscle-specific enzyme creatine kinase (CK) is a marker of striated muscle damage. Avian immunoglobulin (IgY) mediates inflammation and may thereby reduce post-exercise DOMS. Purpose: The aim of this study was to compare the effect of oral IgY and placebo (Pl) on CK, serum relevels, and perceived pain following induced DOMS. Methods: Healthy college-aged females (N=16) were randomly divided into an experimental group (IgY) and a control group (PL). CK serum levels were recorded followed by 14 days of supplementation of either IgY or Pl at the following doses: days 1-2 =4.5 g, days 3-5 =9.0 g, and days 6-14 =13.5 g. Following the 14 d, lower limb DOMS was induced using two methods of resistance training. After 48 hours, subjects reported for a second blood draw. Results: One-way ANOVA resulted in the IgY group posting significantly less (p < 0.05) serum CK than the PL group. Furthermore, the IgY group experienced significantly less post-test perceived soreness than the Pl group. Conclusion: IgY supplementation lessens muscle CK levels and perceived muscle soreness following exercise, possibly due to an anti-inflammatory effect. It was suggested that IgY may serve as a buffer for DOMS thereby allowing the participant to continue vigorous exercise without discomfort.

Keywords: muscle, soreness, damage, serum

Procedia PDF Downloads 197

Authors: Meltem Agar, Maisem Laabei, Hannah Leese, Pedro Estrela

Abstract:

Pathogenic bacteria and the diseases they cause have become a global problem. Their early detection is vital and can only be possible by detecting the bacteria causing the disease accurately and rapidly. Great progress has been made in this field with the use of biosensors. Molecularly imprinted polymers have gain broad interest because of their excellent properties over natural receptors, such as being stable in a variety of conditions, inexpensive, biocompatible and having long shelf life. These properties make molecularly imprinted polymers an attractive candidate to be used in biosensors. In this study it is aimed to produce an aptamer-molecularly imprinted polymer based electrochemical sensor by utilizing the properties of molecularly imprinted polymers coupled with the enhanced specificity offered by DNA aptamers. These ‘apta-MIP’ sensors were used for the detection of Staphylococcus aureus and Escherichia coli. The experimental parameters for the fabrication of sensor were optimized, and detection of the bacteria was evaluated via Electrochemical Impedance Spectroscopy. Sensitivity and selectivity experiments were conducted. Furthermore, molecularly imprinted polymer only and aptamer only electrochemical sensors were produced separately, and their performance were compared with the electrochemical sensor produced in this study. Aptamer-molecularly imprinted polymer based electrochemical sensor showed good sensitivity and selectivity in terms of detection of Staphylococcus aureus and Escherichia coli. The performance of the sensor was assessed in buffer solution and tap water.

Keywords: aptamer, electrochemical sensor, staphylococcus aureus, molecularly imprinted polymer

Procedia PDF Downloads 118

Authors: Sakinul Islam, Nhol Kao, Sati Bhattacharya, Rahul Gupta

Abstract:

Nanocrystalline cellulose (NCC) has been widely used for miscellaneous applications due to its superior properties over other nanomaterials. However, the major problems associated with the production of NCC are long reaction time, low production rate and inefficient process. The mass production of NCC within a short period of time is still a great challenge. The main objective of this study is to produce NCC from rice husk agro waste biomass from a high pressure delignification process (HPDP), followed by bleaching and hydrolysis processes. The HPDP has not been explored for NCC production from rice husk biomass (RHB) until now. In order to produce NCC, powder rice husk (PRH) was placed into a stainless steel reactor at 80 ˚C under 5 bars. Aqueous solution of NaOH (4M) was used for the dissolution of lignin and other amorphous impurities from PRH. After certain experimental times (1h, 3.5h and 6h), bleaching and hydrolysis were carried out on delignified samples. NaOCl (20%) and H2SO4 (4M) solutions were used for bleaching and hydrolysis processes, respectively. The NCC suspension from hydrolysis was sonicated and neutralized by buffer solution for various characterisations. Finally NCC suspension was dried and analyzed by FTIR, XRD, SEM, AFM and TEM. The chemical composition of NCC and PRH was estimated by TAPPI (Technical Association of Pulp and Paper Industry) standard methods to observe the product purity. It was found that, the 6h of the HPDP was more efficient to produce good quality NCC than that at 1h and 3.5h due to low separation of non-cellulosic components from RHB. The analyses indicated the crystallinity of NCC to be 71 %, particle size of 20-50 nm (diameter) and 100-200 nm in length.

Keywords: nanocrystalline cellulose, NCC, high pressure delignification, bleaching, hydrolysis, agro-waste biomass

Procedia PDF Downloads 264

Authors: Piyakajornkul T., Noppiboon S., Hochareon L., Kitsubun P.

Abstract:

Nowadays, DNA vaccines become an interesting subject in the third vaccine generation. The platform of DNA vaccines production has been developed and its downstream process becomes challenging due to the quality of the products in terms of purity and percentage of supercoiled DNA. To overcome these challenges, tangential flow filtration (TFF), which is involved in the purification process, could be used since it provides effective separation of impurity prior to performing further purification steps. However, operating conditions of TFF is varied based on several factors such as sizes of target particle and impurities, a concentration of solution as well as a concentration polarization on the membrane surface. In this study, pVAX1/lacZ was used as a model of TFF optimization in order to prevent a concentration polarization that can lead to the membrane fouling and also minimize a diafiltration volume while maintaining the maximum permeate flux resulting in proper operating times and buffer volume. By using trans membrane pressure (TMP) excursion method, feed flow rates and TMP were varied. The results showed a correlation of permeate flux with TMP where the maximum volume concentration factor reached 2.5 times of the initial volume when feed flow rate and TMP were 7 liters/m²/min and 1 bar, respectively. It was optimal operating conditions before TFF system undergone pressure independent regime. In addition, the diafiltration volume was 14 times of the concentrated volume prior to performing a further anion chromatography process.

Keywords: concentration polarization, DNA vaccines, optimization, permeate flux, pressure dependent, tangential flow filtration (TFF), trans membrane pressure (TMP)

Procedia PDF Downloads 159

Authors: Ravi Patel, Ravisinh Solanki, Dignesh Khunt

Abstract:

A stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method was developed and validated for estimating Nirmatrelvir in its self-emulsifying drug delivery system (SEDDS). The separation of Nirmatrelvir and its degradation products was accomplished by employing an Agilent Zorbax Eclipse plus C18 (250 mm x 4.6 mm, 5 µm) column, through which the mobile phase 5 mM phosphate buffer (pH 4.0) as mobile phase A and Acetonitrile as mobile phase B in a ratio of (40:60 % v/v) was pumped at a flow rate of 1.0 mL/min, through the HPLC system. Chromatographic separation and elution were monitored by a photo-diode array detector at 210 nm. Stress studies have been employed to evaluate this method's ability to indicate stability. Nirmatrelvir was exposed to several stress conditions, such as acid, alkali, oxidative, photolytic, and thermal degradations. Significant degradation was observed during acid and alkali hydrolysis, and the resulting degradation product was successfully separated from the Nirmatrelvir peak, preventing any interference. Furthermore, the primary degradant produced under alkali degradation conditions was identified using UPLC-ESI-TQ-MS/MS. The method was validated in accordance with the International Council on Harmonization (ICH) and found to be selective, precise, accurate, linear, and robust. The apparent permeability of Nirmatrelvir SEDDS was 4.20 ± 0.21×10-6 cm/sec, and the average proportion of free drug recovered was 0.5%. The method developed in this study was feasible and accurate for routine quality control evaluation of Nirmatrelvir SEDDS.

Keywords: Nirmatrelvir, SEDDS, degradation study, HPLC, LC-MS/MS

Procedia PDF Downloads 18

Authors: Sachin Latiyan, T. S. Sampath Kumar, Mukesh Doble

Abstract:

Natural polymer based nanofibrous wound dressings have gained increased attention because of their high surface area, bioactivity, biodegradability and resemblance to extracellular matrix. Agarose (a natural polymer) have been used largely for angiogenesis, cartilage formation and wound healing applications. However, electrospinning of agarose is tedious thereby rendering limited studies on fabrication and evaluation of agarose based nanofibrous wound dressings. Thus, present study focuses on the fabrication of agarose (10% w/v)/ polyvinyl alcohol (12% w/v) based multifunctional nanofibrous scaffolds. Zinc citrate (1, 3 and 5% w/w of the polymer) was added as a potential antibacterial agent to combat wound infections. The fabricated scaffolds exhibit ~500% swelling (in phosphate buffer saline) with enhanced mechanical strength which is suitable for most of the wound healing applications. In vitro studies were found to reveal an increased migration and proliferation of L929 mouse fibroblasts with agarose blends w.r.t to the control. The fabricated dressings were found to be effective against both Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacterial strains. Hence, a multifunctional (as provides effective swelling and mechanical support along with antibacterial property), natural product based, eco-friendly scaffold was successfully fabricated to serve as a potential wound dressing material.

Keywords: antibacterial dressings, benign solvent, nanofibrous agarose, biocompatibility, enhanced swelling and mechanical strength, biopolymeric dressings

Procedia PDF Downloads 93

Authors: Sara Torabi, Sadegh Khazalpour, Mahdi Jamshidi

Abstract:

Nitro aromatic compounds are valuable materials because of their applications in the preparation of chemical intermediates for the synthesis of dyes, plastics, perfumes, energetic materials, and pharmaceuticals. Chemical and electrochemical procedures are reported for nitration of aromatic compounds. Flavonoid derivatives are present in many vegetables and fruits and are constituent of many common pharmaceuticals and dietary supplements. Electrochemistry provides very versatile means for the electrosynthesis, mechanistic and kinetic studies. To the best of our knowledge, and despite the importance of these compounds in numerous scientific fields, there are no reports on the electrochemical nitration of Quercetin derivatives. Herein, we describe a green electrochemical synthesis of a nitro compound. In this work, electrochemical oxidation of Quercetin has been studied in the presence of nitrite ion as a nucleophile in acetate buffer solution (c = 0.2 M, pH = 6.0), by means of cyclic voltammetry and controlled-potential coulometry. The results indicate the participation of produced o-benzoquinones in Michael reaction with nitrite ion (in the divided cell) to form the corresponding nitro diol (EC mechanism). The purity of product and characterization was done using ¹H NMR, ¹³C NMR, FTIR spectroscopic techniques. The presented strategies use a water/ethanol mixture as solvent. Ethanol as cosolvent was also used in the previous studies because of its low cost, safety, easy availability, recyclability, bioproductability, and biodegradability. These strategies represent a one-pot and facile process for the synthesis of nitro compound in high yield and purity under green conditions.

Keywords: electrochemical synthesis, green chemistry, cyclic voltammetry, molecular docking

Procedia PDF Downloads 144

Authors: Anna Wendolowska, Dorota Czyzowska

Abstract:

An intimate relationship is a significant factor that influences romantic partners’ well-being. In the face of stress, avoidant partners often employ a defense-against-intimacy strategy, leading to reduced relationship satisfaction, intimacy, interdependence, and longevity. Dyadic coping can buffer the negative effects of stress on relational satisfaction. Emotional competence mediates the relationship between insecure attachment and intimacy. In the current study, the link between attachment, different forms of dyadic coping, and various aspects of relationship satisfaction was examined. Both partners completed the attachment style questionnaire, the well matching couple questionnaire, and the dyadic coping inventory. The data was analyzed using the actor–partner interdependence model. The results highlighted a negative association between insecure-avoidant attachment style and intimacy. The actor effects of avoidant attachment on relational intimacy for women and for men were significant, whilst the partner effects for both spouses were not significant. The emotion-focused common dyadic coping moderated the relationship between avoidance of attachment and the partner's sense of intimacy. After controlling for the emotion-focused common dyadic coping, the actor effect of attachment on intimacy for men was slightly weaker, and the actor effect for women turned out to be insignificant. The emotion-focused common dyadic coping weakened the negative association between insecure attachment and relational intimacy. The impact of adult attachment and dyadic coping significantly contributes to subjective relational well-being.

Keywords: adult attachment, dyadic coping, relational intimacy, relationship satisfaction

Procedia PDF Downloads 162

Authors: N.Revathy, C.Gomathi

Abstract:

This work aims for a tunable capacitor as a sensor which can vary the control voltage of a voltage control oscillator in a ultra wide band (UWB) transmitter. In this paper power consumption is concentrated. The reason for choosing a capacitive sensing is it give slow temperature drift, high sensitivity and robustness. Previous works report a resistive sensing in a voltage control oscillator (VCO) not aiming at power consumption. But this work aims for power consumption of a capacitive sensing in ultra wide band transmitter. The ultra wide band transmitter to be used is a direct modulation of pulses. The VCO which is the heart of pulse generator of UWB transmitter works on the principle of voltage to frequency conversion. The VCO has and odd number of inverter stages which works on the control voltage input this input is now from a variable capacitor and the buffer stages is reduced from the previous work to maintain the oscillating frequency. The VCO is also aimed to consume low power. Then the concentration in choosing a variable capacitor is aimed. A compact model of a capacitor with the transient characteristics is to be designed with a movable dielectric and multi metal membranes. Previous modeling of the capacitor transient characteristics is with a movable membrane and a fixed membrane. This work aims at a membrane with a wide tuning suitable for ultra wide band transmitter.This is used in this work because a capacitive in a ultra wide transmitter need to be tuned in such a way that all satisfies FCC regulations.

Keywords: capacitive sensing, ultra wide band transmitter, voltage control oscillator, FCC regulation

Procedia PDF Downloads 392

Authors: Herawanti Kumalasari, Raldi Hendro Koestoer, Hayati Sari Hasibuan

Abstract:

Geographically, Indonesia is located in the arc of volcanoes that cause disaster prone one of them is landslide disaster. One of the causes of the landslide is the conversion of land from forest to agricultural land in upland areas and river border that has a steep slope. The study area is located in the highlands with fertile soil conditions, so most of the land is used as agricultural land and plantations. Land use transfer also occurs around the geothermal field in Pangalengan District, West Java Province which will threaten the sustainability of geothermal energy utilization and the safety of the community. The purpose of this research is to arrange the concept of spatial pattern arrangement in the geothermal area based on disaster mitigation. This research method using superimpose analysis. Superimpose analysis to know the basic physical condition of the planned area through the overlay of disaster risk map with the map of the plan of spatial plan pattern of Bandung Regency Spatial Plan. The results of the analysis will then be analyzed spatially. The results have shown that most of the study areas were at moderate risk level. Planning of spatial pattern of existing study area has not fully considering the spread of disaster risk that there are settlement area and the agricultural area which is in high landslide risk area. The concept of the arrangement of the spatial pattern of the study area will use zoning system which is divided into three zones namely core zone, buffer zone and development zone.

Keywords: spatial planning, geothermal, disaster risk, zoning

Procedia PDF Downloads 272

Authors: Junhang Luo

Abstract:

With the transformation of spatial structure and the deepening of urban development, the demand for a better life and the concerns for social resources equities of residents are increasing. As an important social resource, park plays an essential role in building environmentally sustainable cities. Thus, it is important to examine park accessibility for low-income and how it works in relative deprivation, so as to provide all residents with equitable services. Using the network and buffer methods of GIS, this paper analyzes urban park accessibility for low-income residents in Chongqing, China. And then conduct a satisfaction evaluation of park resource accessibility with low-incomes through questionnaire surveys from deprivation dimensions. Results show that the level of park accessibility in Chongqing varies significantly and the degree of relative deprivation is relatively high. Public transportation convenience improves and the number of community park increases contribute positively to improving park accessibility and alleviating the relative deprivation of public resources. Combined with the innovation pattern of social governance in China, it suggests that urban park accessibility needs to be jointly governed and optimized by multiple social resources from the government to the public, and the service efficiency needs the index system and planning standards according to local conditions to improve quality and promote equity. At the same time, building a perfect park system and complete legislation assurance system will also play a positive role in ensuring that all residents can enjoy the urban public space more fairly, especially low-income groups.

Keywords: urban park, accessibility, relative deprivation, GIS network analysis, chongqing

Procedia PDF Downloads 159

Authors: Bernardo Cortez, Florian Peter, Thomas Lausenhammer, Paulo Oliveira

Abstract:

Designing missiles’ autopilot controllers have been a complex task, given the extensive flight envelope and the nonlinear flight dynamics. A solution that can excel both in nominal performance and in robustness to uncertainties is still to be found. While Control Theory often debouches into parameters’ scheduling procedures, Reinforcement Learning has presented interesting results in ever more complex tasks, going from videogames to robotic tasks with continuous action domains. However, it still lacks clearer insights on how to find adequate reward functions and exploration strategies. To the best of our knowledge, this work is a pioneer in proposing Reinforcement Learning as a framework for flight control. In fact, it aims at training a model-free agent that can control the longitudinal non-linear flight dynamics of a missile, achieving the target performance and robustness to uncertainties. To that end, under TRPO’s methodology, the collected experience is augmented according to HER, stored in a replay buffer and sampled according to its significance. Not only does this work enhance the concept of prioritized experience replay into BPER, but it also reformulates HER, activating them both only when the training progress converges to suboptimal policies, in what is proposed as the SER methodology. The results show that it is possible both to achieve the target performance and to improve the agent’s robustness to uncertainties (with low damage on nominal performance) by further training it in non-nominal environments, therefore validating the proposed approach and encouraging future research in this field.

Keywords: Reinforcement Learning, flight control, HER, missile autopilot, TRPO

Procedia PDF Downloads 264

Authors: Sujata Upgupta, Prasoon Kumar Singh

Abstract:

The expansion of large scale coal mining into forest areas is a potential hazard for the local biodiversity and wildlife. The objective of this study is to provide a picture of the threat that coal mining poses to the forests of the East Bokaro landscape. The vulnerable forest areas at risk have been assessed and the priority areas for conservation have been presented. The forested areas at risk in the current scenario have been assessed and compared with the past conditions using classification and buffer based overlay approach. Forest vulnerability has been assessed using an analytical framework based on systematic indicators and composite vulnerability index values. The results indicate that more than 4 km² of forests have been lost from 1973 to 2016. Large patches of forests have been diverted for coal mining projects. Forests in the northern part of the coal field within 1-3 km radius around the coal mines are at immediate risk. The original contiguous forests have been converted into fragmented and degraded forest patches. Most of the collieries are located within or very close to the forests thus threatening the biodiversity and hydrology of the surrounding regions. Based on the vulnerability values estimated, it was concluded that more than 90% of the forested grids in East Bokaro are highly vulnerable to mining. The forests in the sub-districts of Bermo and Chandrapura have been identified as the most vulnerable to coal mining activities. This case study would add to the capacity of the forest managers and mine managers to address the risk and vulnerability of forests at a small landscape level in order to achieve sustainable development.

Keywords: forest, coal mining, indicators, vulnerability

Procedia PDF Downloads 390