Dissertations / Theses on the topic 'Hanging drop'

Thesis advisor: Larry W. McLaughlin<br>The ability to control gene expression has traditionally been pursued at the protein level, using drugs designed to mimic a natural substrate or to disrupt a protein's active site. Traditional drug targeting by competitive and non-competitive inhibitors, however, requires a fairly detailed knowledge of the target protein's three-dimensional structure. More recently, focus has broadened to include alternative methods of genetic control, including the use of single-stranded DNA or RNA probe sequences which control gene expression by targeting the genes themselves. Within the last two decades, peptide nucleic acids (PNAs) – DNA mimics possessing natural bases linked to an N-(2-aminoethyl)-glycine (AEG) backbone – have proven as effective in gene-targeting as traditional synthetic DNA or RNA with the added advantages of tighter binding and greater specificity. Additionally, PNAs are not easily recognized by nucleases, proteases, and peptidases giving them greater resistance to enzyme degradation and making them even more favorable for gene targeting in vivo. Traditional PNA triplexes are composed of two polypyrmidine PNA strands bound to the Watson-Crick and Hoogsteen faces, respectively, of the polypurine strand of target DNA after displacing the polypyrimidine strand of the original DNA duplex. Janus Wedge (JW) residues, on the other hand, utilize unnatural bases linked to the AEG backbone, which are capable of hydrogen bonding to the Watson-Crick faces of both strands of a target DNA duplex. JW triplex formation, then, has a DNA2-PNA stoichiometry, and no Hoogsteen face interactions. The generalization of the DNA duplex targeting strategy by peptide oligomers requires substantial discoveries in the field of PNA research, including an understanding of the three-dimensional structure and folding pattern of these triple-stranded molecules. This report details the crystallization efforts on JW DNA-peptide-DNA triplexes using 11dC811-11T811 target sequences – with and without single base overhangs – and synthetic W8K peptide. Hanging drop vapor diffusion methods showed that while crystal formation was extremely elusive, in narrowing the optimal buffer conditions, 25% PEG concentration was consistently correlated with the most promising crystallization efforts for both the overhanged and non-overhanged sequences<br>Thesis (BS) — Boston College, 2005<br>Submitted to: Boston College. College of Arts and Sciences<br>Discipline: Chemistry<br>Discipline: College Honors Program

Cell-substrate interactions play a key role in the regulation of epithelial cell mechanics. Through a series of studies, we demonstrate how substrate interactions impact both the response to an oncogene and the cellular contractility and organization of a monolayer. We first examine the effects of oncogenic Ras in cells in adherent and suspended states. To accomplish this, we utilized atomic force microscopy and a microfluidic optical stretcher. We found that adherent cells stiffen and suspended cells soften with the expression of constitutively active Ras. The effect on adherent cells was reversed when contractility was inhibited with the ROCK inhibitor Y-27632, resulting in softer cells. These findings suggest that increased ROCK activity as a result of Ras has opposite effects on suspended and adhered cells. In a subsequent study, we examined the effects of a substrate on contracting and relaxing monolayers. We created a new methodology for measuring the mechanodynamics of epithelial monolayers by culturing cells at an air-liquid interface. These model monolayers were grown in the absence of any supporting structures in hanging drops. We found that the direction of strain in the unsupported monolayers was not correlated to nuclear alignment as observed when the monolayers were grown on soft deformable gels. It was also observed that both gel and glass substrates led to the promotion of long-range cell nuclei alignment not seen in the unsupported monolayers. To further characterize the morphology and mechanics of monolayers clusters observed in our experiments, we created a new computational model based on the vertex model. The energy function used in this model takes into account cell-cell and cell-substrate adhesion as well as anisotropic cellular mechanical properties. The results of these simulations suggest that the promotion of long-range alignment on solid substrates were due to cells having anisotropic elastic moduli with global alignment. They also suggest that the alignment observed in monolayers grown on air water interfaces is due to cells having low substrate adhesion and isotropic moduli. Our findings establish the importance of studying epithelial cell mechanics in different states of attachment.

Na presente tese investigou-se a adsorção induzida de um cátion metálico (cádmio(II)) na presença simultânea de dois ligantes (um pseudo-haleto e uma amina ou haleto) na interface eletrodo de mercúrio/solução aquosa, com vistas a uma melhor compreensão desse processo. Os estudos de adsorção foram precedidos por estudos de equilíbrio dos compostos de coordenação formados em solução aquosa. Medidas experimentais foram feitas por potenciometria com eletrodo de amálgama, em força iônica 1,00 M, ajustada com NaCl04. Análise dos dados por programas computacionais adequados levou à quantificação das constantes de estabilidade de 7 espécies binárias e 9 espécies mistas para o sistema Cd(II)/ SCN-/C6H12N4 e de 8 espécies binárias e 6 mistas para o sistema Cd(II)/I-/SCN-, sendo todas estas espécies mononucleares. Para possibilitar um maior grau de automação das medidas cronocoulométricas de adsorção, o eletrodo de gota pendente de mercúrio, anteriormente desenvolvido no IQ-USP, teve seu sistema mecânico aperfeiçoado e passou a ser controlado por computador. O \"software\" se incumbe de calcular e comandar o tempo de abertura da micro-válvula do eletrodo para gerar gotas com a área solicitada. Aproveitando a possibilidade de realizar aquisição (de dados de carga durante e após a formação das gotas de mercúrio, introduziu-se um novo método para corrigir a componente faradaica que interfere nos experimentos de determinação da carga absoluta do eletrodo por extrusão. Após provar o bom funcionamento do método, conseguiu-se a combinação inédita do método de extrusão de gotas com acronocoulometria num único experimento automatizado feito com a mesma gota de mercúrio. Com as informações assim geradas, torna-se possível conhecer e optar entre o potencial e a carga absoluta como parâmetro que define o estado elétrico da interface eletrodo/solução. Com esta combinação de técnicas, foi investigada a adsorção de cádmio induzida por tiocianato e hexametilenotetramina. Este sistema apresentou um interessante comportamento: a hexametilenotetramina, ao contrário do tiocianato, não induz a adsorção de ions cádmio, mas na presença desse pseudo-haleto provoca um intenso reforço na adsorção induzida que pode alcançar um fator de 50 vezes. Também foi possível realizar estimativas sobre as espécies que adsorvem preferencialmente, e da área média ocupada pelos complexos na condição de máxima adsorção. A energia livre de adsorção pode ser estimada pela isoterma de Henry, para baixas concentrações de ligantes. A adsorção de cádmio induzida por iodeto e tiocianato foi investigada de maneira preliminar. Nos estudos deste sistema ficou bem evidenciada a importância da utilização da carga como variável independente, ao invés do potencial.<br>The induced adsorption of a metallic cation (cadmium (II)) in the presence of two ligands, simultaneously (a pseudo-halide and an amine or halide), has been investigated at the mercury electrode/aqueous solution interface with the purpose of gaining a better understanding about such kind of process. The adsorption studies have been preceded by studies of the equilibria of the coordination compounds formed in aqueous solutions. Experimental measurements have been made by potentiometry with an amalgam electrode with 1,00 M ionic strength adjusted with NaClO4. Analysis of the data with adequated software allowed the quantification of the stability constants of 7 binary and 8 mixed species for the system Cd(II)/ SCN-/C6H12N4</sub and 8 binary and 6 mixed species for the system Cd(II)/I-/SCN-, all species being mononuclear. To provide a greater degree of automation of the chronocoulometric measurements of adsorption, the hanging drop mercury electrode, formely developed at the Chemistry Institute-USP, has had its mechanical parts perfectioned and wasput under computer control. A software was written to calculate and control the opening and closure of the micro-solenoid valve of the electrode to generate drops with the required area. A new method for the correction of the faradaic component that interferes with the absolute electrode charge during extrusion experiments was introduced and applied to charge measurements obtained making data aquisition during and shortly after the drop formation. Once proved that the method works, efforts were directed to combine the extrusion with the chronocoulometric measurements in one automated experiment dane with the same mercury drop. As a result, it is now a simple matter to make a choice between potential or charge on the electrode to define the electric state of the electrode/solution interface. With this combination of techniques, the adsorption of cadmium(II) induced by thiocyanate and hexamethylenetetramine has been investigated. An interesting behavior whas observed: the hexamethylenetetramine alone, doesn\'t induce adsorption of cadmium (II) as does the thiocyanate but provides a strong reinforcement (up to a factor of 50) of the induced adsorption when it is present simultaneously with the pseudo-halide. Some estimatives about the species that can adsorb preferentially has been made, as well as an evaluation of the mean area ocupied by the complexes at the maximum adsorption condition. The free energy of adsorption has been calculated, considering an Henry isotherm for the lowest concentration range. Preliminary studies of the induced adsorption of the Cd(II)/I-/SCN- system revealed a slight reinforcement of the adsorption, when both ligands are presente. The iodide always controls the process. The advantage of using the charge in place of potential as the independent variable is evident for this system.

Cette premiere etude d'alliages refractaires dans le tube a chute libre necessite la resolution des problemes d'elaboration des gouttes et de mesure des temperatures associees a la recalescence. Suivant une methode d'ajout, la technique de la goutte pendante permet de couvrir l'ensemble du domaine de composition des systemes re-w et re-ta avec une excellente reproductibilite de la masse et de la temperature initiale des gouttes. Celle-ci s'identifie a la temperature de liquidus de l'alliage ce qui conserve le pouvoir de resolution apporte par l'emploi de la vitesse du refroidissement pour le calcul de la temperature de germination. Une approche pyrometrique fournit la temperature de post-recalescence. Les receptions dans de l'etain conservent des traces des phases metastables rencontrees. De 15 a 65%atw dans le systeme re-w, une germination primaire de la phase metastable c. C. -(w) se produit, la phase d'equilibre apparaissant par transformation a l'etat solide. Un phenomene de double recalescence systematique est mis en evidence entre 65 et 82%atw du a la germination primaire d'une phase metastable a15. Celle-ci est encore detectee dans des gouttes a 40 et 63%atw en presence respectivement de la phase et d'une phase metastable b2. Le calcul du diagramme d'equilibre liquide + a15 rend compte du large domaine de metastabilite de la structure a15 sous reserve de la considerer comme une solution solide a deux sous-reseaux. L'observation d'une phase b2 non stoechiometrique est correlee a la presence d'une phase a15 non stoechiometrique dans ce systeme. L'analyse statistique des evenements de fin de surfusion donne une valeur de la tension d'interface de 0,375 j. M#-#2 pour le compose intermetallique rew#3 de structure a15. La solidification directe de la phase x s'opere dans le systeme re-ta pour une vitesse de trempe associee a la recalescence vingt fois plus faible que celle trouvee par exemple pour la phase metastable c. C. -(w) dans re-w. Le domaine d'existence de la phase x s'etend jusqu'a 70%atta. Entre 83 et 86%atta, une competition de germination se manifeste entre deux phases metastables a15 et. Le trace des courbes de metastabilite rend compte des resultats obtenus. Cette etude s'appuie sur de nombreux calculs de structure electronique qui identifient les phases metastables d'interet (pour l'alliage et les elements purs) et fournissent les grandeurs energetiques permettant de situer leur temperature de fusion. Via les phenomenes de metastabilite, des informations sont tirees sur la sequence assurant le passage de la structure cubique centree a celle de l'hexagonal compact. Cette etude souligne l'importance de la topologie sur la selection des phases. Les tests de microdurete les plus interessants concernant la phase x sursaturee en tantale qui associe durete elevee et ductilite

O tributilestanho é uma espécie química que tem demonstrado ser altamente tóxica para organismos vivos. É utilizado em tintas anti-incrustantes, aplicadas sobretudo no revestimento de cascos de embarcações e outras estruturas marinhas com o objectivo de evitar a deposição de organismos vivos. O tributilestanho, tal como os seus derivados, apresenta um comportamento muito complexo, nomeadamente no que respeita à determinação e quantificação através de métodos electroquímicos. Este trabalho apresenta uma nova abordagem, aliando a voltametria de onda quadrada de redissolução anódica a redes neuronais artificiais. Esta técnica, oriunda da Área Científica da Inteligência Artificial, permite analisar os dados obtidos por voltametria de onda quadrada de redissolução anódica e encontrar um padrão comportamental que permite quantificar o tributilestanho em amostras de água salgada recolhidas no porto de Sesimbra. É uma técnica muito rápida de executar, de baixo custo e que apresenta taxas de recuperação consideravelmente elevadas; ABSTRACT:Tributyltin is a chemical species that has proven to be highly toxic to living organisms. It is used in antifouling paints, especially in boat hulls coating and other marine structures in order to prevent the deposition of living organisms. TBT and its derivatives have a very complex behavior, particularly when it comes to the determination and quantification using electrochemical methods. This project presents a new approach, combining the square wave anodic stripping voltammetry with artificial neural networks. This technique, emanating from the Scientific Area of Artificial Intelligence, enables the analysis of the data obtained by square wave anodic stripping voltammetry and helps in finding a pattern that allows the quantification of Tributyltin in the seawater samples collected at the Sesimbra harbor. This is a very fast and low cost technique which presents considerably high recovery rates.

Bile acids are the end products of cholesterol metabolism and are important biological surfactants. The curved shape of their chains allows the cyclization of molecules, and the formation of a supramolecular structure. The goal of this thesis was to study the electrochemical and adsorption behavior of selected bile acids: lithocholic, deoxycholic and cholic acids. The measurements were carried out in the medium Brittonův-Robinson buffer:methanol in the ratio 9:1 using cyclic voltammetry and AC voltammetry methods and measuring the dependence of the differential capacitance Cd on the applied potential E. A hanging mercury drop electrode was used as a working electrode. The measurements showed that bile acids are adsorbed on the surface of the electrode and organizing themselves in self assembled monolayers (SAM). In our case we have observed formation of 2D condensed layers as specific form of SAM. Transfer techniques were used to demonstrate bile acid adsorption. A study of the behavior of lithocholic acid as a function of different pH values showed that only at pH 10.0 to 12.0 2D 2D condensation occurs, i. e. that at pH values in the range of 2.0 to 9.0 it is another type of adsorption. On AC voltammograms, there are a maximum of two areas in which peaks occur: the first is around -0.2 V and the...

The electrochemic's behaviour of the bile acids (cholic, glycocholic, deoxycholic, ursodeoxycholic and lithocholic acid) was studied on the meniscus-modified silver solid amalgam electrode (m-AgSAE) by differential pulse voltammery. Bile acids provide in the solution of the Britton - Robinson buffer and methanol (9:1) in the pH range 3.0 to 12.0 a cathodal signal in the high negative potentials: cholic acid, deoxycholic acid, ursodeoxycholic acid and lithocholic acid about −1400 mV and glycocholic acid, which alone is the conjugate with glycine, about −1500 mV. Cholic acid, glycocholic acid, deoxycholic acid and ursodeoxycholic acid provide the highest peaks to pH 5.0, approximately in their pKa values. Lithocholic acid provides peaks from pH 7.0. It was demonstrated by the cyclic voltammetry that the electrochemical behavior is influenced by the adsorption of the bile acids to the electrode; presumed reaction at the working electrode - a reduction of a proton of a carboxylic group, is controlled by the diffusion and the process is quasireversible. Utilization of the electrochemical reduction of bile acids for the voltammetric determination does not seem very suitable. It has been proven that the presence of the methanol deteriorates the measuring results for glycocholic acid. In the presence of...

This thesis deals with the study of electrochemical behaviour of dehydrocholic and chenodeoxycholic acid at stationary mercury-based electrodes - hanging mercury dropping electrode and silver solid amalgam electrode modified by a mercury meniscus. This is the first study of electrochemical behaviour of dehydrocholic acid which offers tree peaks with potentials around -1270 mV, -1450 mV and -1800 mV. The last one with the potential -1800 mV is probably the main peak given by reduction of carboxylic group of side chain of steroid skeleton. By cyclic voltammetry was determined that the process is quasireversible and is strongly influenced by adsorption of DHCA on the electrode surface and also by the pH which determines dissociation degree of carboxylic group. In the environment of BR buffer (pH 7.0) - methanol (9:1) was measured concentration dependence by DC, DP and "square-wave" voltammetry. By neither one of these methods have been reached satisfying limits of detection and wide linear dynamic ranges. However, the repeatable signals in wide range of pH can be used for electrochemical characterization of DHCA. CDCA offers in environment 0,04 mol·l-1 borat buffer (pH 9.1) - methanol (9:1) one signal with potential around -1350 mV. Position of this peak on the potential axe and also its height are...

This thesis deals with electrochemical behaviour of tauroursodeoxycholic acid (TUDCA) at silver solid amalgam electrode modified by mercury meniscus (m-AgSAE), polished silver solid amalgam electrode (p-AgSAE) and hanging mercury dropping electrode (HMDE). This thesis is a part of a bigger scientific research that deals with synthesis and characterization of supramolecular systems based on natural steroid compounds and its conjugates. TUDCA offers one reduction peak at m-AgSAE in the environment of Britton - Robinson buffer in range of pH 6.0 - 13.0. The potential of this peak is around −1200 mV. Using cyclic voltammetry was determined that the process on the electrode surface is quasireversible, the reduction is controlled by diffusion and the anodic process is controlled by adsorption. Concentration dependence measured at HMDE by direct current voltammetry in 0.04 mol∙l-1 borat buffer (pH 9.1) is linear in two concentration intervals - 1∙10-3 - 2∙10-4 mol∙l-1 and 1∙10-4 - 8∙10-6 mol∙l-1 of TUDCA. There was no linear dependence between the increase of concentration of TUDCA and the height of the peak obtained on amalgam electrodes by methods DC, DP, cyclic and "square-wave" voltammetry. On HMDE was in several short concentration intervals measured by a CV method a linear dependence of cathodic and...

The main objective of this Diploma Thesis is studying the possibility of detection of nitro derivatives of aromatic hydrocarbons by using chemically modified working electrodes in electrochemical analysis. Compounds analyzed in this work were 1-nitronaphthalene and 6- nitrochrysene - these compounds were chosen on previous experience with surface modified working electrodes (the chemical/physical properties of these substances). All the analyses were performed by using differential pulse voltammetry (DPV) on hanging mercury drop electrode (HMDE) in classic three-electrode system. The original premise of chemical modification of electrode surface is the change of its selectivity towards different compounds - meaning the change of the electrochemical response. The modification agent used in this thesis is 1-octanethiol. Its thiol group offers great potential of strong interaction with the metal surface of the mercury electrode. Voltammetric responses of both compounds were recorded in acidic, neutral and basic solutions (pH=2, pH=7, pH=12) including the responses when the electrode surface was modified. Next objective was the optimization of the modification procedure itself (transferring of the mercury drop from one media to another without damaging or tearing it off from the capillary orifice)....

This Diploma Thesis is focused on investigation of the voltammetric behavior of the explosive penthrite (pentaerythritol tetranitrate) (PETN) and on searching for optimum conditions for its determination using differential pulse voltammetry (DPV) at a hanging mercury drop electrode (HMDE) (this part of the Diploma Thesis follows the topic investigated in the author's Bachelor Thesis; in this Diploma Thesis, the results of the Bachelor thesis are applied to the determination of PETN in environmental matrices) and at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE) and on investigation of voltammetric behavior in real samples (deionized, drinking, and river water). For investigating the behavior of PETN in aqueous-methanolic media on the m-AgSAE, mixtures of Britton-Robinson buffers (BR-buffers) and methanol in various volume ratios were used. In non-aqueous media, methanolic solutions of tetramethylammonium bromide (TMAB), tetrabutylammonium iodide (TBAI), and tetrabutylammonium chloride (TBAC) were used as supporting electrolytes. Firstly, the influence of pH (in the range of 2 - 13) and methanol content (10 - 90 vol. %) on voltammetric responses of PETN was studied. The best response was obtained at each of those pH values at the volume ratio of BR-buffer - methanol of 1:9 or...

In this Diploma Thesis, an interaction of genotoxic environmental pollutant 2-nitrofluorene with a double-stranded calf thymus DNA has been studied using a hanging mercury drop electrode (HMDE) as an electrochemical sensor. Two types of DNA damage were investigated and electrochemically detected (using cyclic voltammetry and differential pulse voltammetry): (i) The DNA damage caused by the direct interaction with 2-nitrofluorene and (ii) the DNA damage caused by short-lived radicals generated by the electrochemical reduction of the nitro group in 2-nitrofluorene. For the study of direct interaction, HMDE was modified by DNA and the interaction of DNA with 2-nitrofluorene was studied, after their incubation, right at the HMDE surface (adsorptive transfer stripping technique) or the DNA was preincubated with 2-nitrofluorene and, subsequently, the interaction was studied voltammetrically (DNA titration technique). Using both detection techniques, the formation of DNA - 2-nitrofluorene complex was observed and the mutual interaction was interpreted as an intercalation between the DNA base pairs, although such interaction was not clearly confirmed by UV-visible absorption spectroscopy. An electrostatic binding of 2-nitrofluorene on DNA sugar-phosphate backbone was partially formed at low concentrations of...

The presented diploma thesis is devoted to study of determination of 4-nitrobiphenyl (4-NBP) in model river water by DC voltammetry (DCV) and differential pulse voltammetry (DPV) at m-AgSAE (diploma thesis relates to bachelor thesis, in it DCV and DPV methods for determination of 4-NBP in deionized water were developed). Limit of determination (LOD) is 2·10-7 mol·l-1 by DCV and 4·10-7 mol·l-1 by DPV. The adsorptive stripping DPV (AdSDPV) technique was tested to archive lower LOD the AdSDPV, optimal conditions were not found. Electrochemical behavior of 2-nitrobiphenyl (2-NBP) at m-AgSAE was studied. Optimal conditions for its determination by both techniques DCV and DPV were found in methanol-0,01 mol·l-1 LiOH (1:9) and in this medium peak current dependence in 1·10-7 mol·l-1 to 1·10-4 mol·l-1 2-NBP (LOD ≈ 2·10-7 mol·l-1 by DCV and 1·10-7 mol·l-1 by DPV) concentration range was measured. Developed methods were successfully tested for determination of 2-NBP in drinking and river water samples. For both mediums obtained LODs were 2·10-7 mol·l-1 by DCV and 1·10-7 mol·l-1 by DPV. AdSDPV technique was unsuccessfully tested to achieve lower LOD. Optimal conditions for simultaneous determination of 4-NBP and 2-NBP by DPV technique at m-AgSAE were found: methanol-0,25 mol·l-1 acetate buffer (pH = 6,0) 3:7....

Presented Diploma Thesis is focused on electroanalytical determination of the herbicide Aclonifen, which belongs to a group of herbicides derived from diphenylether. These herbicides are used due to their effects - inhibition of protoporfyrinogen oxidase synthesis and also inhibition of biosynthesis of carotenoids. Due to its widespread use in agricultural, toxical effects on humans and because of its negative impacts particularly on aquatic ecosystems there is a need for methods capable of determining the presence of Aclonifen in the environment to monitor its ecological impacts. Optimal conditions for the determination of Aclonifen have been investigated in BR buffer - methanolic solution and in BR buffer. Electrochemical behavior of the substance has been studied using direct current voltammetry (DCV), differential pulse voltammetry (DPP) and adsorptive stripping voltammetry (AdSV) on the hanging mercury drop electrode (HMDE) and by using TAST polarography and differential pulse polarography (DPP) on the dropping mercury electrode (DME). UV/VIS spectrophotometric detection was used for comparison to electrochemical detection. Used wavelengths were 308 nm and 388 nm. For electrochemical determination of Aclonifen the above mentioned techniques were used and following results were obtained: DCV...

Presented Diploma Thesis is focused on electroanalytical determination of genotoxic 4-nitroindane, one of the nitrated polycyclic aromatic hydrocarbons (NPAHs). A hydrocarbon indane (a component of petrol) is a precursor of 4-nitroindane. NPAHs are produced all above by combustion processes in gasoline and diesel engines. It has been shown that NPAHs can be many times more mutagenic or carcinogenic than their parent PAHs, so the analysis of these dangerous pollutants becomes important for modern environmental analytical chemistry. Optimal conditions for determination of 4-nitroindane have been investigated in buffered water-methanolic solutions and electrochemical transformations of 4-nitroindane have been studied by DC voltammetry (DCV), differential pulse voltammetry (DPV) and cyclic voltammetry at a hanging mercury drop electrode (HMDE) and at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE). For voltammetric determination of 4-nitroindane, the following techniques were used: DCV (limit of quantification (LQ) ~ 7. 10-8 mol. l-1 ), DPV (LQ ~ 1. 10-7 mol. l-1 ) and adsorptive stripping voltammetry (AdSV; LQ ~ 7. 10-9 mol. l-1 ) at HMDE, and DCV (LQ ~ 1. 10-7 mol. l-1 ) and DPV (LQ ~ 1. 10-7 mol l-1 ) at m-AgSAE. The applicability of the newly developed...