scholarly journals ELECTRON MICROSCOPIC AND KINETIC STUDIES DEALING WITH AN ARTIFICIAL ENZYME MEMBRANE APPLICATION TO A CYTOCHEMICAL MODEL WITH THE HORSERADISH PEROXIDASE-3,3'-DIAMINOBENZIDINE SYSTEM

1974 ◽  
Vol 22 (11) ◽  
pp. 1048-1059 ◽  
Author(s):  
J. N. BARBOTIN ◽  
D. THOMAS

A new colorimetric method for measuring horseradish peroxidase (HRP) activity, in solution, with 3,3'-diaminobenzidine (DAB) as the hydrogen donor is presented. The same method was also used to test HRP activity when immobilized in an artificial proteic membrane. The reaction was shown to be of first order in both cases. Direct spectrophotometric measurements dealing with HRP membrane show that the total amount of insoluble polymer produced does not increase linearly with the quantity of enzyme introduced in the membrane. Moreover, a study of the DAB permeability through membranes with or without peroxidase activity was performed to allow the study of diffusion limitations. Membranes have also been studied by electron microscopy. We have especially visualized the existence of product concentration profiles inside the membrane. The results show that in the membrane there is no geometrical similarity between the distribution profiles of the enzyme and of the insoluble product.

Author(s):  
A. Angel ◽  
K. Miller ◽  
V. Seybold ◽  
R. Kriebel

Localization of specific substances at the ultrastructural level is dependent on the introduction of chemicals which will complex and impart an electron density at specific reaction sites. Peroxidase-antiperoxidase(PAP) methods have been successfully applied at the electron microscopic level. The PAP complex is localized by addition of its substrate, hydrogen peroxide and an electron donor, usually diaminobenzidine(DAB). On oxidation, DAB forms an insoluble polymer which is able to chelate with osmium tetroxide becoming electron dense. Since verification of reactivity is visual, discrimination of reaction product from osmiophillic structures may be difficult. Recently, x-ray microanalysis has been applied to examine cytochemical reaction precipitates, their distribution in tissues, and to study cytochemical reaction mechanisms. For example, immunoreactive sites labelled with gold have been ascertained by means of x-ray microanalysis.


1992 ◽  
Vol 57 (9) ◽  
pp. 1951-1959 ◽  
Author(s):  
Madlene L. Iskander ◽  
Samia A. El-Abbady ◽  
Alyaa A. Shalaby ◽  
Ahmed H. Moustafa

The reactivity of the base induced cyclodimerization of 1-(6-arylpyridazin-3-yl)-3-oxidopyridinium chlorides in a pericyclic process have been investigated kinetically at λ 380 nm. The reaction was found to be second order with respect to the liberated betaine and zero order with respect to the base. On the other hand dedimerization (monomer formation) was found to be first order. It was shown that dimerization is favoured at low temperature, whereas dedimerization process is favoured at relatively high temperature (ca 70 °C). Solvent effects on the reaction rate have been found to follow the order ethanol > chloroform ≈ 1,2-dichloroethane. Complete dissociation was accomplished only in 1,2-dichloroethane at ca 70 °C. The thermodynamic activation parameters have been calculated by a standard method. Thus, ∆G# has been found to be independent on substituents and solvents. The high negative values of ∆S# supports the cyclic transition state which is in favour with the concerted mechanism. MO calculations using SCF-PPP approximation method indicated low HOMO-LUMO energy gap of the investigated betaines.


1987 ◽  
Vol 35 (8) ◽  
pp. 909-916 ◽  
Author(s):  
G D Gagne ◽  
M F Miller

We describe an artificial substrate system for optimization of labeling parameters in electron microscope immunocytochemical studies. The system involves use of blocks of glutaraldehyde-polymerized BSA into which a desired antigen is incorporated by a simple soaking procedure. The resulting antigen-impregnated artificial substrate can then be fixed and embedded identically to a piece of tissue. The BSA substrate can also be dried and then sectioned for immunolabeling with or without chemical fixation and without exposing the antigen to dehydrating agents and embedding resins. The effects of various fixation and embedding procedures can thus be evaluated separately. Other parameters affecting immunocytochemical labeling, such as antibody and conjugate concentration, can also be evaluated. We used this system, along with immunogold labeling, to determine quantitatively the optimal fixation and embedding conditions for labeling of hepatitis B surface antigen (HbsAg), human IgG, and horseradish peroxidase. Using unfixed and unembedded HBsAg, we were able to detect antigen concentrations below 20 micrograms/ml. We have shown that it is not possible to label HBsAg within resin-embedded cells using conventional aldehyde fixation protocols and polyclonal antibodies.


2017 ◽  
Vol 8 (7) ◽  
pp. 4833-4839 ◽  
Author(s):  
Lei Zhang ◽  
Fengjiao Ma ◽  
Jianping Lei ◽  
Jintong Liu ◽  
Huangxian Ju

A hemin-based enzymatic network is constructed with high catalytic activity that is comparable to natural horseradish peroxidaseviatriple signal amplification.


2008 ◽  
Vol 32 (2) ◽  
pp. 533-540 ◽  
Author(s):  
Vladimir Antônio Silva ◽  
Giuliano Marchi ◽  
Luiz Roberto Guimarães Guilherme ◽  
José Maria de Lima ◽  
Francisco Dias Nogueira ◽  
...  

Kinetic studies on soil potassium release can contribute to a better understanding of K availability to plants. This study was conducted to evaluate K release rates from the whole soil, clay, silt, and sand fractions of B-horizon samples of a basalt-derived Oxisol and a sienite-derived Ultisol, both representative soils from coffee regions of Minas Gerais State, Brazil. Potassium was extracted from each fraction after eight different shaking time periods (0-665 h) with either 0.001 mol L-1 citrate or oxalate at a 1:10 solid:solution ratio. First-order, Elovich, zero-order, and parabolic diffusion equations were used to parameterize the time dependence of K release. For the Oxisol, the first-order equation fitted best to the experimental data of K release, with similar rates for all fractions and independent of the presence of citrate or oxalate in the extractant solution. For all studied Ultisol fractions, in which K release rates increased when extractions were performed with citrate solution, the Elovich model described K release kinetics most adequately. The highest potassium release rate of the Ultisol silt fraction was probably due to the transference of "non-exchangeable" K to the extractant solution, whereas in the Oxisol exchangeable potassium represented the main K source in all studied fractions.


2018 ◽  
Vol 156 ◽  
pp. 02012 ◽  
Author(s):  
Mardiah ◽  
Rif’an Fathoni ◽  
Pratiwi Pudyaningtyas ◽  
Hamdania Gamu ◽  
Rinaldy

High Consumption of paper, bring the impact of the waste paper itself. And the utilization of the paper is limited to recycled products and crafts, whereas paper such as newspaper still contains cellulose that can be potential to be used as a heavy metal adsorbent. In this study, newspaper was dissolved in sodium bicarbonate to reduce various impurities and then was reacted with citric acid (CA). The modified adsorbent was characterized by FTIR and was tested for adsorb Cu(II) in artificial solution. After adsorption process, the solution was filtered and analysed using Atomic Absorption Spectrophotometer (AAS). The adsorption experimental data was fitted to Langmuir, Freundlich, Tempkin, and Dubinin-Radushkevich for equilibrium model and was fitted to pseudo first order reaction and pseudo second order reaction for kinetic studies. The result showed that CA-modification newspaper able to remove heavy metals Cu(II) in solution.


2020 ◽  
Vol 42 ◽  
pp. e9
Author(s):  
Alex Leandro Andrade de Lucena ◽  
Daniella Carla Napoleão ◽  
Hélder Vinícius Carneiro da Silva ◽  
Rayany Magali da Rocha Santana ◽  
Beatriz Galdino Ribeiro ◽  
...  

The existence of pharmaceuticals in nature is a growing environmental problem, turning necessary the use of efficient treatments for the degradation of these substances, as the advanced oxidation processes (AOPs). In this work the AOPs UV/H2O2 and photo-Fenton were applied to degrade the pharmaceuticals lamivudine and zidovudine in an aqueous solution using a bench reactor, composed of three UV-C lamps. It was verified that the UV/H2O2 process presented a degradation of 97.33 ± 0.14% for lamivudine and 93.90 ± 0.33% for zidovudine, after 180 min of treatment and for an initial concentratin of each pharmaceutical of  5 mg.L-1 and [H2O2] of 600 mg.L-1.  A methodology by artificial neural networks (ANNs) was used to model the photocatalytic process, with the MLP 7-23-2 ANN representing it well, and determining the relative importance (%) of each of the input variables for the pharmaceutical’s degradation process. Kinetic studies for the pharmaceutical degradation and the conversion of organic matter showed good adjustments to the pseudo first-order models with R2 raging from 0.9705 to 0.9980. Toxicity assays for the before treatment solution indicated that the seeds Lactuca sativa and Portulaca grandiflora showed growth inhibition whereas the post-treatment solution inhibited only the growth of Lactuca sativa.


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