Search results for: release kinetics
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1840

Search results for: release kinetics

640 Process Evaluation for a Trienzymatic System

Authors: C. Müller, T. Ortmann, S. Scholl, H. J. Jördening

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Multienzymatic catalysis can be used as an alternative to chemical synthesis or hydrolysis of polysaccharides for the production of high value oligosaccharides from cheap resources such as sucrose. However, development of multienzymatic processes is complex, especially with respect to suitable conditions for enzymes originating from different organisms. Furthermore, an optimal configuration of the catalysts in a reaction cascade has to be found. These challenges can be approached by design of experiments. The system investigated in this study is a trienzymatic catalyzed reaction which results in laminaribiose production from sucrose and comprises covalently immobilized sucrose phosphorylase (SP), glucose isomerase (GI) and laminaribiose phosphorylase (LP). Operational windows determined with design of experiments and kinetic data of the enzymes were used to optimize the enzyme ratio for maximum product formation and minimal production of byproducts. After adjustment of the enzyme activity ratio to 1: 1.74: 2.23 (SP: LP: GI), different process options were investigated in silico. The considered options included substrate dependency, the use of glucose as co-substrate and substitution of glucose isomerase by glucose addition. Modeling of batch operation in a stirred tank reactor led to yields of 44.4% whereas operation in a continuous stirred tank reactor resulted in product yields of 22.5%. The maximum yield in a bienzymatic system comprised of sucrose phosphorylase and laminaribiose phosphorylase was 67.7% with sucrose and different amounts of glucose as substrate. The experimental data was in good compliance with the process model for batch operation. The continuous operation will be investigated in further studies. Simulation of operational process possibilities enabled us to compare various operational modes regarding different aspects such as cost efficiency, with the minimum amount of expensive and time-consuming practical experiments. This gives us more flexibility in process implementation and allows us, for example, to change the production goal from laminaribiose to higher oligosaccharides.

Keywords: design of experiments, enzyme kinetics, multi-enzymatic system, in silico process development

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639 Freedom of Information and Freedom of Expression

Authors: Amin Pashaye Amiri

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Freedom of information, according to which the public has a right to have access to government-held information, is largely considered as a tool for improving transparency and accountability in governments, and as a requirement of self-governance and good governance. So far, more than ninety countries have recognized citizens’ right to have access to public information. This recognition often took place through the adoption of an act referred to as “freedom of information act”, “access to public records act”, and so on. A freedom of information act typically imposes a positive obligation on a government to initially and regularly release certain public information, and also obliges it to provide individuals with information they request. Such an act usually allows governmental bodies to withhold information only when it falls within a limited number of exemptions enumerated in the act such as exemptions for protecting privacy of individuals and protecting national security. Some steps have been taken at the national and international level towards the recognition of freedom of information as a human right. Freedom of information was recognized in a few countries as a part of freedom of expression, and therefore, as a human right. Freedom of information was also recognized by some international bodies as a human right. The Inter-American Court of Human Rights ruled in 2006 that Article 13 of the American Convention on Human Rights, which concerns the human right to freedom of expression, protects the right of all people to request access to government information. The European Court of Human Rights has recently taken a considerable step towards recognizing freedom of information as a human right. However, in spite of the measures that have been taken, public access to government information is not yet widely accepted as an international human right. The paper will consider the degree to which freedom of information has been recognized as a human right, and study the possibility of widespread recognition of such a human right in the future. It will also examine the possible benefits of such recognition for the development of the human right to free expression.

Keywords: freedom of information, freedom of expression, human rights, government information

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638 A pH-Activatable Nanoparticle Self-Assembly Triggered by 7-Amino Actinomycin D Demonstrating Superior Tumor Fluorescence Imaging and Anticancer Performance

Authors: Han Xiao

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The development of nanomedicines has recently achieved several breakthroughs in the field of cancer treatment; however, the biocompatibility and targeted burst release of these medications remain a limitation, which leads to serious side effects and significantly narrows the scope of their applications. The self-assembly of intermediate filament protein (IFP) peptides was triggered by a hydrophobic cation drug 7-amino actinomycin D (7-AAD) to synthesize pH-activatable nanoparticles (NPs) that could simultaneously locate tumors and produce antitumor effects. The designed IFP peptide included a target peptide (arginine–glycine–aspartate), a negatively charged region, and an α-helix sequence. It also possessed the ability to encapsulate 7-AAD molecules through the formation of hydrogen bonds and hydrophobic interactions by a one-step method. 7-AAD molecules with excellent near-infrared fluorescence properties could be target delivered into tumor cells by NPs and released immediately in the acidic environments of tumors and endosome/lysosomes, ultimately inducing cytotoxicity by arresting the tumor cell cycle with inserted DNA. It is noteworthy that the IFP/7-AAD NPs tail vein injection approach demonstrated not only high tumor-targeted imaging potential, but also strong antitumor therapeutic effects in vivo. The proposed strategy may be used in the delivery of cationic antitumor drugs for precise imaging and cancer therapy.

Keywords: 7-amino actinomycin D, intermediate filament protein, nanoparticle, tumor image

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637 The Limits to Self-Defense Claims in Case of Domestic Violence Homicides

Authors: Maria Elisabete Costa Ferreira

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Domestic violence is a serious social issue in which victims are mostly women. Domestic violence develops in cycles, starting with the building of tension, passing through the incident of abuse and ending with reconciliation, also known as honeymoon. As time goes by, the shorter these phases become, and the greater and more severe the attacks, rarely leading to the death of the victim of abuse. Sometimes, the victim stops the abuse by killing the aggressor, usually after the immediate aggression has taken place. This poses an important obstacle to the claim of self-defense by the victim of domestic violence pending trial for the homicide of her long-time abuser. The main problem with self-defense claims in such cases is that the law requires the act of aggression to be present or imminent (imminent threat or immediate danger) so that it permits the victim to take her defense into her own hands. If the episode of aggression has already taken place, this general requirement for the admissibility of self-defense is not satisfied. This paper sheds new light on the concept of the actuality of the aggression, understanding that, since domestic violence is a permanent offense, for as long as the victim stays under the domain of the aggressor, imminent threat will be present, allowing the self-defense claim of a woman who kills her abuser in such circumstances to be admissible. An actualist interpretation of the requirement of the necessity of the means used in self-defense will be satisfied when evaluated from the subjective perspective of the intimate partner victim. Necessity will be satisfied if it is reasonable for the victim to perceive the use of lethal force as the only means to release herself from the abuser.

Keywords: domestic violence, homicide, self-defense, imminent threat, necessity of lethal force

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636 Extracellular Phytase from Lactobacillus fermentum spp KA1: Optimization of Enzyme Production and Its Application for Improving the Nutritional Quality of Rice Bran

Authors: Neha Sharma, Kanthi K. Kondepudi, Naveen Gupta

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Phytases are phytate specific phosphatases catalyzing the step-wise dephosphorylation of phytate, which acts as an anti-nutritional factor in food due to its strong binding capacity to minerals. In recent years microbial phytases have been explored for improving nutritional quality of food. But the major limitation is acceptability of phytases from these microorganisms. Therefore, efforts are being made to isolate organisms which are generally regarded as safe for human consumption such as Lactic Acid Bacteria (LAB). Phytases from these organisms will have an edge over other phytase sources due to its probiotic attributes. Only few LAB have been reported to give phytase activity that too is generally seen as intracellular. LAB producing extracellular phytase will be more useful as it can degrade phytate more effectively. Moreover, enzyme from such isolate will have application in food processing also. Only few species of Lactobacillus producing extracellular phytase have been reported so far. This study reports the isolation of a probiotic strain of Lactobacillus fermentum spp KA1 which produces extracellular phytase. Conditions for the optimal production of phytase have been optimized and the enzyme production resulted in an approximately 13-fold increase in yield. The phytate degradation potential of extracellular phytase in rice bran has been explored and conditions for optimal degradation were optimized. Under optimal conditions, there was 43.26% release of inorganic phosphate and 6.45% decrease of phytate content.

Keywords: Lactobacillus, phytase, phytate reduction, rice bran

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635 Potential and Techno-Economic Analysis of Hydrogen Production from Portuguese Solid Recovered Fuels

Authors: A. Ribeiro, N. Pacheco, M. Soares, N. Valério, L. Nascimento, A. Silva, C. Vilarinho, J. Carvalho

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Hydrogen will play a key role in changing the current global energy paradigm, associated with the high use of fossil fuels and the release of greenhouse gases. This work intended to identify and quantify the potential of Solid Recovered Fuels (SFR) existing in Portugal and project the cost of hydrogen, produced through its steam gasification in different scenarios, associated with the size or capacity of the plant and the existence of carbon capture and storage (CCS) systems. Therefore, it was performed a techno-economic analysis simulation using an ASPEN base model, the H2A Hydrogen Production Model Version 3.2018. Regarding the production of SRF, it was possible to verify the annual production of more than 200 thousand tons of SRF in Portugal in 2019. The results of the techno-economic analysis simulations showed that in the scenarios containing a high (200,000 tons/year) and medium (40,000 tons/year) amount of SFR, the cost of hydrogen production was competitive concerning the current prices of hydrogen. The results indicate that scenarios 1 and 2, which use 200,000 tons of SRF per year, have lower hydrogen production values, 1.22 USD/kg H2 and 1.63 USD/kg H2, respectively. The cost of producing hydrogen without carbon capture and storage (CCS) systems in an average amount of SFR (40,000 tons/year) was 1.70 USD/kg H2. In turn, scenarios 5 (without CCS) and 6 (with CCS), which use only 683 tons of SFR from urban sources, have the highest costs, 6.54 USD/kg H2 and 908.97 USD/kg H2, respectively. Therefore, it was possible to conclude that there is a huge potential for the use of SRF for the production of hydrogen through steam gasification in Portugal.

Keywords: gasification, hydrogen, solid recovered fuels, techno-economic analysis, waste-to-energy

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634 Thorium Extraction with Cyanex272 Coated Magnetic Nanoparticles

Authors: Afshin Shahbazi, Hadi Shadi Naghadeh, Ahmad Khodadadi Darban

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In the Magnetically Assisted Chemical Separation (MACS) process, tiny ferromagnetic particles coated with solvent extractant are used to selectively separate radionuclides and hazardous metals from aqueous waste streams. The contaminant-loaded particles are then recovered from the waste solutions using a magnetic field. In the present study, Cyanex272 or C272 (bis (2,4,4-trimethylpentyl) phosphinic acid) coated magnetic particles are being evaluated for the possible application in the extraction of Thorium (IV) from nuclear waste streams. The uptake behaviour of Th(IV) from nitric acid solutions was investigated by batch studies. Adsorption of Thorium (IV) from aqueous solution onto adsorbent was investigated in a batch system. Adsorption isotherm and adsorption kinetic studies of Thorium (IV) onto nanoparticles coated Cyanex272 were carried out in a batch system. The factors influencing Thorium (IV) adsorption were investigated and described in detail, as a function of the parameters such as initial pH value, contact time, adsorbent mass, and initial Thorium (IV) concentration. Magnetically Assisted Chemical Separation (MACS) process adsorbent showed best results for the fast adsorption of Th (IV) from aqueous solution at aqueous phase acidity value of 0.5 molar. In addition, more than 80% of Th (IV) was removed within the first 2 hours, and the time required to achieve the adsorption equilibrium was only 140 minutes. Langmuir and Frendlich adsorption models were used for the mathematical description of the adsorption equilibrium. Equilibrium data agreed very well with the Langmuir model, with a maximum adsorption capacity of 48 mg.g-1. Adsorption kinetics data were tested using pseudo-first-order, pseudo-second-order and intra-particle diffusion models. Kinetic studies showed that the adsorption followed a pseudo-second-order kinetic model, indicating that the chemical adsorption was the rate-limiting step.

Keywords: Thorium (IV) adsorption, MACS process, magnetic nanoparticles, Cyanex272

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633 The Effects of Zinc Oxide Nanoparticles Loaded with Indole-3-Acetic Acid and Indole-3-Butyric Acid on in vitro Rooting of Apple Microcuttings

Authors: Shabnam Alizadeh, Hatice Dumanoglu

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Plant tissue culture is a substantial plant propagation technique for mass clonal production throughout the year, regardless of time in fruit species. However, the rooting achievement must be enhanced in the difficult-to-root genotypes. Classical auxin applications in clonal propagation of these genotypes are inadequate to solve the rooting problem. Nanoparticles having different physical and chemical properties from bulk material could enhance the rooting success of controlled release of these substances when loaded with auxin due to their ability to reach the active substance up to the target cells as a carrier system.The purpose of this study is to investigate the effects of zinc oxide nanoparticles loaded with indole-3-acetic acid (IAA-nZnO) and indole-3-butyric acid (IBA-nZnO) on in vitro rooting of microcuttings in a difficult-to-root apple genotype (Malus domestica Borkh.). Rooting treatments consisted of IBA or IAA at concentrations of 0.5, 1.0, 2.0, 3.0 mg/L; nZnO, IAA-nZnO and IBA-nZnO at doses of 0.0, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 mg/L were used. All components were added to the Murashige and Skoog (MS) basal medium at strength ½ with 2% sucrose and 0.7% agar before autoclaving. In the study, no rooting occurred in control and nZnO applications. Especially, 1.0 mg/L and 2.0 mg/L IBA-nZnO nanoparticle applications (containing 0.5 mg/L and 0.9 mg/L IBA), respectively with rooting rates of 40.3% and 70.4%, rooting levels of 2.0±0.4 and 2.3±0.4, 2.6±0.7 and 2.5±0.6 average root numbers and 20.4±1.6 mm and 20.2±3.4 mm average root lengths put forward as effective applications.

Keywords: Auxin, Malus, nanotechnology, zinc oxide nanoparticles

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632 Transfer Function Model-Based Predictive Control for Nuclear Core Power Control in PUSPATI TRIGA Reactor

Authors: Mohd Sabri Minhat, Nurul Adilla Mohd Subha

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The 1MWth PUSPATI TRIGA Reactor (RTP) in Malaysia Nuclear Agency has been operating more than 35 years. The existing core power control is using conventional controller known as Feedback Control Algorithm (FCA). It is technically challenging to keep the core power output always stable and operating within acceptable error bands for the safety demand of the RTP. Currently, the system could be considered unsatisfactory with power tracking performance, yet there is still significant room for improvement. Hence, a new design core power control is very important to improve the current performance in tracking and regulating reactor power by controlling the movement of control rods that suit the demand of highly sensitive of nuclear reactor power control. In this paper, the proposed Model Predictive Control (MPC) law was applied to control the core power. The model for core power control was based on mathematical models of the reactor core, MPC, and control rods selection algorithm. The mathematical models of the reactor core were based on point kinetics model, thermal hydraulic models, and reactivity models. The proposed MPC was presented in a transfer function model of the reactor core according to perturbations theory. The transfer function model-based predictive control (TFMPC) was developed to design the core power control with predictions based on a T-filter towards the real-time implementation of MPC on hardware. This paper introduces the sensitivity functions for TFMPC feedback loop to reduce the impact on the input actuation signal and demonstrates the behaviour of TFMPC in term of disturbance and noise rejections. The comparisons of both tracking and regulating performance between the conventional controller and TFMPC were made using MATLAB and analysed. In conclusion, the proposed TFMPC has satisfactory performance in tracking and regulating core power for controlling nuclear reactor with high reliability and safety.

Keywords: core power control, model predictive control, PUSPATI TRIGA reactor, TFMPC

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631 Preparation and Characterization of Water-in-Oil Nanoemulsion of 5-Fluorouracil to Enhance Skin Permeation for Treatment of Skin Diseases.

Authors: P. S. Rajinikanth, Shobana Mariappan, Jestin Chellian

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The objective of the study was to prepare and characterize a water-in-oil nano emulsion of 5-Fluorouracil (5FU) to enhance the skin penetration. The present study describes a nano emulsion of 5FU using Capyrol PGMC, Transcutol HP and PEG 400 as oil, surfactant and co-surfactant, respectively. The optimized formulations were further evaluated for heating cooling cycle, centrifugation studies, freeze thaw cycling, particle size distribution and zeta potential in order to confirm the stability of the optimized nano emulsions. The in-vitro characterization results showed that the droplets of prepared formulation were ~100 nm with ± 15 zeta potential. In vitro skin permeation studies was conducted in albino mice skin. Significant increase in permeability parameters was also observed in nano emulsion formulations (P<0.05). The steady-state flux (Jss), enhancement ration and permeability coefficient (Kp) for optimized nano emulsion formulation (FU2, FU1, 1:1 S mix were found to be 24.21 ±2.45 μg/cm2/h, 3.28±0.87 & 19.52±1.87 cm/h, respectively), which were significant compared with conventional gel. The in vitro and in vivo skin deposition studies in rat indicated that the amount of drug deposited from the nano emulsion (292.45 µg/cm2) in skin was significant (P<0.05) an increased as compared to a conventional 5FU gel (121.42 µg/cm2). The skin irritation study using rat skin showed that the mean irritation index of the nano emulsion reduced significantly (P<0.05) as compared with conventional gel contain 1% 5FU. The results from this study suggest that a water-in-oil nano emulsion could be safely used to promote skin penetration of 5FU following topical application.

Keywords: nano emulsion, controlled release, 5 fluorouracil, skin penetration, skin irritation

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630 Investigation of Flame and Soot Propagation in Non-Air Conditioned Railway Locomotives

Authors: Abhishek Agarwal, Manoj Sarda, Juhi Kaushik, Vatsal Sanjay, Arup Kumar Das

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Propagation of fire through a non-air conditioned railway compartment is studied by virtue of numerical simulations. Simultaneous computational fire dynamics equations, such as Navier-Stokes, lumped species continuity, overall mass and energy conservation, and heat transfer are solved using finite volume based (for radiation) and finite difference based (for all other equations) solver, Fire Dynamics Simulator (FDS). A single coupe with an eight berth occupancy is used to establish the numerical model, followed by the selection of a three coupe system as the fundamental unit of the locomotive compartment. Heat Release Rate Per Unit Area (HRRPUA) of the initial fire is varied to consider a wide range of compartmental fires. Parameters, such as air inlet velocity relative to the locomotive at the windows, the level of interaction with the ambiance and closure of middle berth are studied through a wide range of numerical simulations. Almost all the loss of lives and properties due to fire breakout can be attributed to the direct or indirect exposure to flames or to the inhalation of toxic gases and resultant suffocation due to smoke and soot. Therefore, the temporal stature of fire and smoke are reported for each of the considered cases which can be used in the present or extended form to develop guidelines to be followed in case of a fire breakout.

Keywords: fire dynamics, flame propagation, locomotive fire, soot flow pattern, non-air-conditioned coaches

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629 Development and Characterization of Soya Phosphatidylcholine Complex of Coumestans from Eclipta alba for the Management of Hepatotoxicity

Authors: Abhishek Kumar Jain, Anki Jain, Yuvraj Singh Dangi, Brajesh Kumar Tiwari

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The plant Eclipta alba Hassk. (Family: Compositae) contains coumestans (wedelolactone and demethyl wedelolactone) used in liver disorders. The objective of the present investigation was to develop a formulation of these coumestans in combination with the soya phosphatidylcholine (PC), in order to overcome the limitation of absorption and to investigate the protective effect of coumestans–phosphatidylcholine complex (C-PC) on carbon tetrachloride induced acute liver damage in rats. Methanolic extract (ME) of the whole plant of Eclipta alba was fractionated with water and then with ehylacetate. Coumestans were characterized in the ethylacetate fraction of methanolic extract (EFME). The C-PC was prepared by dissolving EFME and PC in 1:1 ratio in dichloromethane and heating at 60°C for 2 h. The C-PC was characterized by DSC and FTIR spectroscopy. In vitro drug release from EFME and C-PC through egg membrane was measured using UV-Visible spectrophotometer. The hepatoprotective activity of C-PC (equivalent to 5.35 and 10.7 mg/kg body weight of EFME), ME 250 mg/kg and EFME 5.35 mg/kg was evaluated by measuring various enzymes level. C-PC significantly provided better protection to the liver by restoring the enzyme levels of SGPT, SGOT, ALP and total billirubin with respect to carbon tetrachloride (CCl4) treated group (P < 0.001). Histopathological studies were also performed. The C-PC provided better protection to rat liver than ME and EFME at similar doses as well as shown significant regeneration of hepatocytes, central vein, intact cytoplasm, and nucleus.

Keywords: hepatotoxicity, wedelolactone, soya phosphatidylcholine, eclipta alba

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628 Synthesis and Cytotoxic Activity of New Quinazolinone-Based Compounds against Human Breast Cancer Cell Line MCF-7

Authors: Maryam Zahedifard, Fadhil Lafta Faraj, Maryam Hajrezaie, Nazia Abdul Majid, Mahmood Ameen Abdulla, Hapipah Mohd Ali

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In the current study, we prepared two new quinazoline schiff bases through condensation reaction of 2-aminobenzhydrazide with 5-bromosalicylaldehyde and 3-methoxy-5-bromosalicylaldehyde. The chemical structures of both newly synthesized compounds (1 and 2) were confirmed by FT-IR and X-ray crystallography studies. The cytotoxic effect of compounds was investigated against MCF-7 human breast cancer cells. MTT results showed that (1) and (2) decreased the viability of MCF-7 cells in a time-dependent manner, exhibiting an IC50 value of 3.23 ± 0.28 µg/mL and 3.41 ± 0.34 µg/mL, respectively, after a 72-hours treatment period. In contrast, they did not show significant anti-proliferative effect towards MCF-10A normal breast cells and WRL-68 normal liver cells. We found a perturbation in mitochondrial membrane potential and increased cytochrome c release from the mitochondria to the cytosol, suggesting an activation of apoptosis by compounds, which was confirmed by activation of the initiator caspase-9 and the executioner caspases-3/7. (1) was also able to trigger extrinsic pathway via activation of caspase-8 and inhibition of NF-κB translocation. The acute toxicity test showed no toxicity effect of the compounds in rats. Our results showed that the selected synthesized compounds are highly potent to induce apoptosis in MCF-7 cells via either intrinsic or extrinsic mitochondrial pathway.

Keywords: Quinazoline Schiff base, apoptosis, MCF-7 human breast cancer cell line, caspase, NF-κB translocation

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627 Ab-initio Calculations on the Mechanism of Action of Platinum and Ruthenium Complexes in Phototherapy

Authors: Eslam Dabbish, Fortuna Ponte, Stefano Scoditti, Emilia Sicilia, Gloria Mazzone

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The medical techniques based on the use of light for activating the drug are occupying a prominent place in the cancer treatment due to their selectivity that contributes to reduce undesirable side effects of conventional chemotherapy. Among these therapeutic treatments, photodynamic therapy (PDT) and photoactivated chemotherapy (PACT) are emerging as complementary approaches for selective destruction of neoplastic tissue through direct cellular damage. Both techniques rely on the employment of a molecule, photosensitizer (PS), able to absorb within the so-called therapeutic window. Thus, the exposure to light of otherwise inert molecules promotes the population of excited states of the drug, that in PDT are able to produce the cytotoxic species, such as 1O2 and other ROS, in PACT can be responsible of the active species release or formation. Following the success of cisplatin in conventional treatments, many other transition metal complexes were explored as anticancer agents for applications in different medical approaches, including PDT and PACT, in order to improve their chemical, biological and photophysical properties. In this field, several crucial characteristics of candidate PSs can be accurately predicted from first principle calculations, especially in the framework of density functional theory and its time-dependent formulation, contributing to the understanding of the entire photochemical pathways involved which can ultimately help in improving the efficiency of a drug. A brief overview of the outcomes on some platinum and ruthenium-based PSs proposed for the application in the two phototherapies will be provided.

Keywords: TDDFT, metal complexes, PACT, PDT

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626 Insight into the Electrocatalytic Activities of Nitrogen-Doped Graphyne and Graphdiyne Families: A First-Principles Study

Authors: Bikram K. Das, Kalyan K. Chattopadhyay

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The advent of 2-D materials in the last decade has induced a fresh spur of growth in fuel cell technology as these materials have some highly promising traits that can be exploited to felicitate Oxygen Reduction Reaction (ORR) in an efficient way. Among the various 2-D carbon materials, graphyne (Gy) and graphdiyne (Gdy)1 with their intrinsic non-uniform charge distribution holds promises in this purpose and it is expected2 that substitutional Nitrogen (N) doping could further enhance their efficiency. In this regard, dispersive force corrected density functional theory is used to map the oxygen reduction reaction (ORR) kinetics of five different kinds of N doped graphyne and graphdiyne systems (namely αGy, βGy, γGy, RGy and 6,6,12Gy and Gdy) in alkaline medium. The best doping site for each of the Gy/ Gdy system is determined comparing the formation energies of the possible doping configurations. Similarly, the best di-oxygen (O₂) adsorption sites for the doped systems are identified by comparing the adsorption energies. O₂ adsorption on all N doped Gy/ Gdy systems is found to be energetically favorable. ORR on a catalyst surface may occur either via the Eley-Rideal (ER) or the Langmuir–Hinschelwood (LH) pathway. Systematic studies performed on the considered systems reveal that all of them favor the ER pathway. Further, depending on the nature of di-oxygen adsorption ORR can follow either associative or dissociative mechanism; the possibility of occurrence of both the mechanisms is tested thoroughly for each N doped Gy/ Gdy. For the ORR process, all the Gy/Gdy systems are observed to prefer the efficient four-electron pathway but the expected monotonically exothermic reaction pathway is found only for N doped 6,6,12Gy and RGy following the associative pathway and for N doped βGy, γGy and Gdy following the dissociative pathway. Further computation performed for these systems reveals that for N doped 6,6,12Gy, RGy, βGy, γGy and Gdy the overpotentials are 1.08 V, 0.94 V, 1.17 V, 1.21 V and 1.04 V respectively depicting N doped RGy is the most promising material, to carry out ORR in alkaline medium, among the considered ones. The stability of the ORR intermediate states with the variation of pH and electrode potentials is further explored with Pourbiax diagrams and the activities of these systems in the alkaline medium are compared with the prior reported B/N doped identical systems for ORR in an acidic medium in terms of a common descriptor.

Keywords: graphdiyne, graphyne, nitrogen-doped, ORR

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625 Impact of Fluid Flow Patterns on Metastable Zone Width of Borax in Dual Radial Impeller Crystallizer at Different Impeller Spacings

Authors: A. Čelan, M. Ćosić, D. Rušić, N. Kuzmanić

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Conducting crystallization in an agitated vessel requires a proper selection of mixing parameters that would result in a production of crystals of specific properties. In dual impeller systems, which are characterized by a more complex hydrodynamics due to the possible fluid flow interactions, revealing a clear link between mixing parameters and crystallization kinetics is still an open issue. The aim of this work is to establish this connection by investigating how fluid flow patterns, generated by two impellers mounted on the same shaft, reflect on metastable zone width of borax decahydrate, one of the most important parameters of the crystallization process. Investigation was carried out in a 15-dm3 bench scale batch cooling crystallizer with an aspect ratio (H/T) equal to 1.3. For this reason, two radial straight blade turbines (4-SBT) were used for agitation. Experiments were conducted at different impeller spacings at the state of complete suspension. During the process of an unseeded batch cooling crystallization, solution temperature and supersaturation were continuously monitored what enabled a determination of the metastable zone width. Hydrodynamic conditions in the vessel achieved at different impeller spacings investigated were analyzed in detail. This was done firstly by measuring the mixing time required to attain the desired level of homogeneity. Secondly, fluid flow patterns generated in a described dual impeller system were both photographed and simulated by VisiMix Turbulent software. Also, a comparison of these two visualization methods was performed. Experimentally obtained results showed that metastable zone width is definitely affected by the hydrodynamics in the crystallizer. This means that this crystallization parameter can be controlled not only by adjusting the saturation temperature or cooling rate, as is usually done, but also by choosing a suitable impeller spacing that will result in a formation of crystals of wanted size distribution.

Keywords: dual impeller crystallizer, fluid flow pattern, metastable zone width, mixing time, radial impeller

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624 Effects of Aromatase Inhibitor (Fadrozole) Induced Sex-Reversal in Chicken (Gimmizah strain) on Morphology

Authors: Hatem Shreha

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Aromatase inhibitors administered before sexual differentiation of the gonads can induce sex reversal in female chickens (phenotypic male). To analyze the process of sex reversal, we have followed for several months the changes induced by Fadrozole, a nonsteroidal aromatase inhibitor on the morphology of female sex-reversed and female sex-reversed supplemented with L-tyrosine which was previously shown to stimulate the release of Gn Rh. Fadrozole (1mg/egg) was injected into eggs on day four of incubation, phenotypic males and phenotypic males treated with L-tyrosine and males hatched from eggs injected Fadrozole were sacrificed by slaughtering at 16 weeks old and the remaining chicks were sacrificed at 28 weeks old. Both sexes from control chickens were sacrificed at the same age (16 &28 weeks). Hatchability, behavior, body weight, shank length, comb weight, testes weight, blood cells count and wattle weight of sex reversal were tested at 16 and 28 weeks. The results showed that body weight, comb weight, wattles weight and shank length of sex-reversed females were significantly different from control female. Behavior of phenotypic males and phenotypic males fed on L- tyrosine showed aggressive sexual behavior like that of control males and absence of laying behavior. In conclusion our results confirm that Fedrazole injection in eggs before sex differentiation produce a male behavior and morphological index of male in female chicken.

Keywords: sex reversal, fadrozole, phenotypic male, L- tyrosine

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623 Gas Chromatography Coupled to Tandem Mass Spectrometry and Liquid Chromatography Coupled to Tandem Mass Spectrometry Qualitative Determination of Pesticides Found in Tea Infusions

Authors: Mihai-Alexandru Florea, Veronica Drumea, Roxana Nita, Cerasela Gird, Laura Olariu

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The aim of this study was to investigate the residues of pesticide found in tea water infusions. A multi-residues method to determine 147 pesticides has been developed using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) procedure and dispersive solid phase extraction (d-SPE) for the cleanup the pesticides from complex matrices such as plants and tea. Sample preparation was carefully optimized for the efficient removal of coextracted matrix components by testing more solvent systems. Determination of pesticides was performed using GC-MS/MS (100 of pesticides) and LC-MS/MS (47 of pesticides). The selected reaction monitoring (SRM) mode was chosen to achieve low detection limits and high compounds selectivity and sensitivity. Overall performance was evaluated and validated according to DG-SANTE Guidelines. To assess the pesticide residue transfer rate (qualitative) from dried tea in infusions the samples (tea) were spiked with a mixture of pesticides at the maximum residues level accepted for teas and herbal infusions. In order to investigate the release of the pesticides in tea preparations, the medicinal plants were prepared in four ways by variation of water temperature and the infusion time. The pesticides from infusions were extracted using two methods: QuEChERS versus solid-phase extraction (SPE). More that 90 % of the pesticides studied was identified in infusion.

Keywords: tea, solid-phase extraction (SPE), selected reaction monitoring (SRM), QuEChERS

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622 Potassium Fertilization Improves Rice Yield in Aerobic Production System by Decreasing Panicle Sterility

Authors: Abdul Wakeel, Hafeez Ur Rehman, Muhammad Umair Mubarak

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Rice is the second most important staple food in Pakistan after wheat. It is not only a healthy food for the people of all age groups but also a source of foreign exchange for Pakistan. Instead of bright history for Basmati rice production, we are suffering from multiple problems reducing yield and quality as well. Rice lodging and water shortage for an-aerobic rice production system is among major glitches of it. Due to water shortage an-aerobic rice production system has to be supplemented or replaced by aerobic rice system. Aerobic rice system has been adopted for production of non-basmati rice in many parts of the world. Also for basmati rice, significant efforts have been made for aerobic rice production, however still has to be improved for effective recommendations. Among two major issues for aerobic rice, weed elimination has been solved to great extent by introducing suitable herbicides, however, low yield production due weak grains and panicle sterility is still elusive. It has been reported that potassium (K) has significant role to decrease panicle sterility in cereals. Potassium deficiency is obvious for rice under aerobic rice production system due to lack of K gradient coming with irrigation water and lowered indigenous K release from soils. Therefore it was hypothesized that K application under aerobic rice production system may improve the rice yield by decreasing panicle sterility. Results from pot and field experiments confirm that application of K fertilizer significantly increased the rice grain yield due to decreased panicle sterility and improving grain health. The quality of rice was also improved by K fertilization.

Keywords: DSR, Basmati rice, aerobic, potassium

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621 Origanum vulgare as a Possible Modulator of Testicular Endocrine Function in Mice

Authors: Eva Tvrdá, Barbora Babečková, Michal Ďuračka, Róbert Kirchner, Július Árvay

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This study was designed to assess the in vitro effects of Origanum vulgare L. (oregano) extract on the testicular steroidogenesis. We focused on identifying major biomolecules present in the oregano extract, as well as to investigate its in vitro impact on the secretion of cholesterol, testosterone, dehydroepiandrosterone and androstenedione by murine testicular fragments. The extract was subjected to high performance liquid chromatography (HPLC) which identified cyranosid, daidzein, thymol, rosmarinic and trans-caffeic acid among the predominant biochemical components of oregano. For the in vitro experiments, testicular fragments from 20 sexually mature Institute of Cancer Research (ICR) mice were incubated in the absence (control group) or presence of the oregano extract at selected concentrations (10, 100 and 1000 μg/mL) for 24 h. Cholesterol levels were quantified using photometry and the hormones were assessed by ELISA (Enzyme-Linked Immunosorbent Assay). Our data revealed that the release of cholesterol and androstenedione (but not dehydroepiandrosterone and testosterone) by the testicular fragments was significantly impacted by the oregano extract in a dose-dependent fashion. Supplementation of the extract resulted in a significant decline of cholesterol (P < 0.05 in case of 100 μg/mL; P < 0.01 with respect 100 μg/mL extract), as well as androstenedione (P < 0.01 with respect to 100 and 1000 μg/mL extract). Our results suggest that the biomolecules present in Origanum vulgare L. could exhibit a dose-dependent impact on the secretion of male steroids, playing a role in the regulation of testicular steroidogenesis.

Keywords: mice, Origanum vulgare L., steroidogenesis, testes

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620 Stress Evaluation at Lower Extremity during Walking with Unstable Shoe

Authors: Sangbaek Park, Seungju Lee, Soo-Won Chae

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Unstable shoes are known to strengthen lower extremity muscles and improve gait ability and to change the user’s gait pattern. The change in gait pattern affects human body enormously because the walking is repetitive and steady locomotion in daily life. It is possible to estimate the joint motion including joint moment, force and inertia effect using kinematic and kinetic analysis. However, the change of internal stress at the articular cartilage has not been possible to estimate. The purpose of this research is to evaluate the internal stress of human body during gait with unstable shoes. In this study, FE analysis was combined with motion capture experiment to obtain the boundary condition and loading condition during walking. Motion capture experiments were performed with a participant during walking with normal shoes and with unstable shoes. Inverse kinematics and inverse kinetic analysis was performed with OpenSim. The joint angle and muscle forces were estimated as results of inverse kinematics and kinetics analysis. A detailed finite element (FE) lower extremity model was constructed. The joint coordinate system was added to the FE model and the joint coordinate system was coincided with OpenSim model’s coordinate system. Finally, the joint angles at each phase of gait were used to transform the FE model’s posture according to actual posture from motion capture. The FE model was transformed into the postures of three major phases (1st peak of ground reaction force, mid stance and 2nd peak of ground reaction force). The direction and magnitude of muscle force were estimated by OpenSim and were applied to the FE model’s attachment point of each muscle. Then FE analysis was performed to compare the stress at knee cartilage during gait with normal shoes and unstable shoes.

Keywords: finite element analysis, gait analysis, human model, motion capture

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619 Highly Active, Non-Platinum Metal Catalyst Material as Bi-Functional Air Cathode in Zinc Air Battery

Authors: Thirupathi Thippani, Kothandaraman Ramanujam

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Current research on energy storage has been paid to metal-air batteries, because of attractive alternate energy source for the future. Metal – air batteries have the probability to significantly increase the power density, decrease the cost of energy storage and also used for a long time due to its high energy density, low-level pollution, light weight. The performance of these batteries mostly restricted by the slow kinetics of the oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) on cathode during battery discharge and charge. The ORR and OER are conventionally carried out with precious metals (such as Pt) and metal oxides (such as RuO₂ and IrO₂) as catalysts separately. However, these metal-based catalysts are regularly undergoing some difficulties, including high cost, low selectivity, poor stability and unfavorable to environmental effects. So, in order to develop the active, stable, corrosion resistance and inexpensive bi-functional catalyst material is mandatory for the commercialization of zinc-air rechargeable battery technology. We have attempted and synthesized non-precious metal (NPM) catalysts comprising cobalt and N-doped multiwalled carbon nanotubes (N-MWCNTs-Co) were synthesized by the solid-state pyrolysis (SSP) of melamine with Co₃O₄. N-MWCNTs-Co acts as an excellent electrocatalyst for both the oxygen reduction reaction (ORR) and the oxygen evolution reaction (OER), and hence can be used in secondary metal-air batteries and in unitized regenerative fuel cells. It is important to study the OER and ORR at high concentrations of KOH as most of the metal-air batteries employ KOH concentrations > 4M. In the first 16 cycles of the zinc-air battery while using N-MWCNTs-Co, 20 wt.% Pt/C or 20 wt.% IrO₂/C as air electrodes. In the ORR regime (the discharge profile of the zinc-air battery), the cell voltage exhibited by N-MWCNTs-Co was 44 and 83 mV higher (based on 5th cycle) in comparison to of 20 wt.% Pt/C and 20 wt.% IrO₂/C respectively. To demonstrate this promise, a zinc-air battery was assembled and tested at a current density of 0.5 Ag⁻¹ for charge-discharge 100 cycles.

Keywords: oxygen reduction reaction (ORR), oxygen evolution reaction(OER), non-platinum, zinc air battery

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618 Time Temperature Dependence of Long Fiber Reinforced Polypropylene Manufactured by Direct Long Fiber Thermoplastic Process

Authors: K. A. Weidenmann, M. Grigo, B. Brylka, P. Elsner, T. Böhlke

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In order to reduce fuel consumption, the weight of automobiles has to be reduced. Fiber reinforced polymers offer the potential to reach this aim because of their high stiffness to weight ratio. Additionally, the use of fiber reinforced polymers in automotive applications has to allow for an economic large-scale production. In this regard, long fiber reinforced thermoplastics made by direct processing offer both mechanical performance and processability in injection moulding and compression moulding. The work presented in this contribution deals with long glass fiber reinforced polypropylene directly processed in compression moulding (D-LFT). For the use in automotive applications both the temperature and the time dependency of the materials properties have to be investigated to fulfill performance requirements during crash or the demands of service temperatures ranging from -40 °C to 80 °C. To consider both the influence of temperature and time, quasistatic tensile tests have been carried out at different temperatures. These tests have been complemented by high speed tensile tests at different strain rates. As expected, the increase in strain rate results in an increase of the elastic modulus which correlates to an increase of the stiffness with decreasing service temperature. The results are in good accordance with results determined by dynamic mechanical analysis within the range of 0.1 to 100 Hz. The experimental results from different testing methods were grouped and interpreted by using different time temperature shift approaches. In this regard, Williams-Landel-Ferry and Arrhenius approach based on kinetics have been used. As the theoretical shift factor follows an arctan function, an empirical approach was also taken into consideration. It could be shown that this approach describes best the time and temperature superposition for glass fiber reinforced polypropylene manufactured by D-LFT processing.

Keywords: composite, dynamic mechanical analysis, long fibre reinforced thermoplastics, mechanical properties, time temperature superposition

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617 Real-Time Kinetic Analysis of Labor-Intensive Repetitive Tasks Using Depth-Sensing Camera

Authors: Sudip Subedi, Nipesh Pradhananga

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The musculoskeletal disorders, also known as MSDs, are common in construction workers. MSDs include lower back injuries, knee injuries, spinal injuries, and joint injuries, among others. Since most construction tasks are still manual, construction workers often need to perform repetitive, labor-intensive tasks. And they need to stay in the same or an awkward posture for an extended time while performing such tasks. It induces significant stress to the joints and spines, increasing the risk of getting into MSDs. Manual monitoring of such tasks is virtually impossible with the handful of safety managers in a construction site. This paper proposes a methodology for performing kinetic analysis of the working postures while performing such tasks in real-time. Skeletal of different workers will be tracked using a depth-sensing camera while performing the task to create training data for identifying the best posture. For this, the kinetic analysis will be performed using a human musculoskeletal model in an open-source software system (OpenSim) to visualize the stress induced by essential joints. The “safe posture” inducing lowest stress on essential joints will be computed for different actions involved in the task. The identified “safe posture” will serve as a basis for real-time monitoring and identification of awkward and unsafe postural behaviors of construction workers. Besides, the temporal simulation will be carried out to find the associated long-term effect of repetitive exposure to such observed postures. This will help to create awareness in workers about potential future health hazards and encourage them to work safely. Furthermore, the collected individual data can then be used to provide need-based personalized training to the construction workers.

Keywords: construction workers’ safety, depth sensing camera, human body kinetics, musculoskeletal disorders, real time monitoring, repetitive labor-intensive tasks

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616 The Relationship between Trace Elements in Groundwater Linked to a History of Volcanic Activity in La Pampa and Buenos Aires Provinces, Argentina

Authors: Maisarah Jaafar, Neil I. Ward

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Volcanic and geothermal activity can result in the release of arsenic (As), manganese (Mn), iron, selenium (Se), molybdenum (Mo) and uranium (U) into natural waters. Several studies have reported high levels of these elements in surface and groundwater in Argentina. The main focus has been on As associated with volcanic ash deposits. This study reports the trace element levels of groundwater from an agricultural region of south-eastern La Pampa and southern Buenos Aires provinces, Argentina which have reported high levels of human health problems (bone/teeth disorders, depression, arthritis, etc). Fifty-eight groundwater samples were collected from wells adjacent to Ruta 35 and an Agilent 7700x inductively coupled plasma mass spectrometer (ICP-MS) were used for total elemental analysis. Physicochemical analysis confirmed pH range of 7.05-8.84 and variable conductivity (988-3880 µS/cm) with total dissolved solid content of 502-1989 mg/l. The majority water samples are in an oxidizing environment (Eh= 45-146 mV). Total As levels ranged from (µg/l): 13.08 – 319.4 for La Pampa (LP) and 39.6 – 189.4 for Buenos Aires (BA); all above the WHO Guideline for Drinking Water, 10 µg/l As. Interestingly, Mo (LP: 1.85 – 85.39 µg/l; BA: 4.61– 55.55 µg/l;), Se (LP: 1.2 – 16.59 µg/l; BA: 0.3– 6.94 µg/l;) and U (LP: 1.85 – 85.39 µg/l; BA: 4.61– 55.55 µg/l;) levels are lower than reported values for northern La Pampa. Inter-elemental correlation displayed positive statistically significant between As-Mo, A-Se, As-U while negative statistically significant between As-Mn and As-Fe. This confirms that the source of the trace element is similar to that reported for other region of Argentina, namely volcanic ash deposition.

Keywords: Argentina, groundwater, trace element, volcanic activity

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615 Preservation of High Quality Fruit Products: Microwave Freeze Drying as a Substitute for the Conventional Freeze Drying Process

Authors: Sabine Ambros, Ulrich Kulozik

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Berries such as blue- and raspberries belong to the most valuable fruits. To preserve the characteristic flavor and the high contents of vitamins and anthocyanins, the very sensitive berries are usually dried by lyophilization. As this method is very time- and energy-consuming, the dried fruit is extremely expensive. However, healthy snack foods are growing in popularity. Especially dried fruit free of any additives or additional sugar are more and more asked for. To make these products affordable, the fruits have to be dried by a method that is more energy-efficient than freeze drying but reveals the same high product quality. The additional insertion of microwaves to a freeze drying process was examined in this work to overcome the inconveniences of freeze drying. As microwaves penetrate the product volumetrically, sublimation takes place simultaneously all over the product and leads to a many times shorter process duration. A range of microwave and pressure settings was applied to find the optimum drying condition. The influence of the process parameters microwave power and chamber pressure on drying kinetics, product temperature and product quality was investigated to find the best condition for an energy-efficient process with high product quality. The product quality was evaluated by rehydration capacitiy, crispiness, shrinkage, color, vitamin C content and antioxidative capacity. The conclusion could be drawn that microwave freeze dried berries were almost equal to freeze dried fruit in all measured quality parameters or even could overcome it. Additionally, sensory evaluations could confirm the analytical studies. Drying time could be reduced by more than 75% at much lower energy consumption rates. Thus, an energy-efficient and cost saving method compared to the conventional freeze drying technique for the gentle production of tasty fruit or vegetable snacks has been found. This technique will make dried high-quality snacks available for many of consumers.

Keywords: blueberries, freeze drying, microwave freeze drying, process parameters, product quality

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614 Presence of High Concentrations of Toxic Metals from the Collected Soil Samples Due to Excessive E-Waste Burning in the Various Areas of Moradabad City, U.P India

Authors: Aprajita Singh, Anamika Tripathi, Surya P. Dwivedi

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Moradabad is a small town in the Northern area of Uttar Pradesh, India. It is situated on the bank of river Ramganga which is also known as ‘Brass City of India’. There is eventually increase in the environmental pollution due to uncontrolled and inappropriate e-waste burning (recycling) activities which have been reported in many areas of Moradabad. In this paper, analysis of toxic heavy metals, causing pollution to the surrounding environment released from the e-waste burning and much other recycling process. All major e-waste burning sites are situated on the banks of the river which is burned in open environmental conditions. Soil samples were collected from seven (n=3) different sites including control site, after digestion of soil samples using triacid mixture, analysis of different toxic metals (Pb, Ar, Hg, Cd, Cr, Cu, Zn, Fe, and Ni) has been carried out with the help of instrument ICP-AAS. After the study, the outcome is that the soil of those areas contains a relatively high level of the toxic metals in order of Cu>Fe>Pb>Cd>Cr>Zn>Ar>Hg. The concentration of Cd, Pb, Cr, Ar and Zn (the majority of samples experimentally proved) exceeded the maximum standard level of WHO. Sequentially this study showed that uncontrolled e-waste processing operations caused serious pollution to local soil and release of toxic metals in the environment is also causing adverse effect on the health of people living in the nearby areas making them more prone to various harmful diseases.

Keywords: brass city, environment pollution, e-waste, toxic heavy metals

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613 Synthesis and Characterization of Carboxymethyl Cellulose-Chitosan Based Composite Hydrogels for Biomedical and Non-Biomedical Applications

Authors: K. Uyanga, W. Daoud

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Hydrogels have attracted much academic and industrial attention due to their unique properties and potential biomedical and non-biomedical applications. Limitations on extending their applications have resulted from the synthesis of hydrogels using toxic materials and complex irreproducible processing techniques. In order to promote environmental sustainability, hydrogel efficiency, and wider application, this study focused on the synthesis of composite hydrogels matrices from an edible non-toxic crosslinker-citric acid (CA) using a simple low energy processing method based on carboxymethyl cellulose (CMC) and chitosan (CSN) natural polymers. Composite hydrogels were developed by chemical crosslinking. The results demonstrated that CMC:2CSN:CA exhibited good performance properties and super-absorbency 21× its original weight. This makes it promising for biomedical applications such as chronic wound healing and regeneration, next generation skin substitute, in situ bone regeneration and cell delivery. On the other hand, CMC:CSN:CA exhibited durable well-structured internal network with minimum swelling degrees, water absorbency, excellent gel fraction, and infra-red reflectance. These properties make it a suitable composite hydrogel matrix for warming effect and controlled and efficient release of loaded materials. CMC:2CSN:CA and CMC:CSN:CA composite hydrogels developed also exhibited excellent chemical, morphological, and thermal properties.

Keywords: citric acid, fumaric acid, tartaric acid, zinc nitrate hexahydrate

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612 Functional Expression and Characterization of a Novel Indigenous Endo-Beta 1,4- Glucanase from Apis mellifera

Authors: Amtul Jamil Sami

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Apis mellifera is an insect of immense economic importance lives on rich carbohydrate diet including cellulose, nectar, honey and pollen. The carbohydrate metabolism in A mellifera has not been understood fully, as there are no data available, on the functional expression of cellulase gene. The cellulose hydrolyzing enzyme is required for the digestion of pollen cellulose wall, to release the important nutrients (amino acids, minerals, vitamins etc.) from the pollen. A dissection of Apis genome had revealed that there is one gene present for the expression of endo-beta-1,4-glucanase, for cellulose hydrolysis. In the presented work, functional expression of endo-beta-1,4 glucanase gene is reported. Total soluble proteins of the honey bee were isolated and were tested cellulose hydrolyzing enzyme activity, using carboxy-methyl cellulose, as a substrate. A mellifera proteins were able to hydrolyze carboxy-methyl cellulose, confirming its endo- type mode of action. Endo beta-1,4 glucanase enzyme was only present in the gut tissues, no activity was detected in the salivary glands. The pH optima of the enzyme were in the acidic pH range of 4-5-5-0, indicating its metabolic role in the acidic stomach of A mellifera. The reported enzyme is unique, as endo-beta- 1,4 glucanase was able to generate non reducing sugar, as an end product. The results presented, are supportive to the information that the honey bee is capable of producing its novel endo-beta-1,4 glucanase. Further it could be helpful, in understanding, the carbohydrate metabolism in A mellifera.

Keywords: honey bees, Endo-beta 1, 4- glucanase, Apis mellifera, functional expression

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611 Rapid Formation of Ortho-Boronoimines and Derivatives for Reversible and Dynamic Bioconjugation Under Physiological Conditions

Authors: Nicholas C. Rose, Christopher D. Spicer

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The regeneration of damaged or diseased tissues would provide an invaluable therapeutic tool in biological research and medicine. Cells must be provided with a number of different biochemical signals in order to form mature tissue through complex signaling networks that are difficult to recreate in synthetic materials. The ability to attach and detach bioactive proteins from material in an iterative and dynamic manner would therefore present a powerful way to mimic natural biochemical signaling cascades for tissue growth. We propose to reversibly attach these bioactive proteins using ortho-boronoimine (oBI) linkages and related derivatives formed by the reaction of an ortho-boronobenzaldehyde with a nucleophilic amine derivative. To enable the use of oBIs for biomaterial modification, we have studied binding and cleavage processes with precise detail in the context of small molecule models. A panel of oBI complexes has been synthesized and screened using a novel Förster resonance energy transfer (FRET) assay, using a cyanine dye FRET pair (Cy3 and Cy5), to identify the most reactive boron-aldehyde/amine nucleophile pairs. Upon conjugation of the dyes, FRET occurs under Cy3 excitation and the resultant ratio of Cy3:Cy5 emission directly correlates to conversion. Reaction kinetics and equilibria can be accurately quantified for reactive pairs, with dissociation constants of oBI derivatives in water (KD) found to span 9-orders of magnitude (10⁻²-10⁻¹¹ M). These studies have provided us with a better understanding of oBI linkages that we hope to exploit to reversibly attach bioconjugates to materials. The long-term aim of the project is to develop a modular biomaterial platform that can be used to help combat chronic diseases such as osteoarthritis, heart disease, and chronic wounds by providing cells with potent biological stimuli for tissue engineering.

Keywords: dynamic, bioconjugation, bornoimine, rapid, physiological

Procedia PDF Downloads 98