Effects of carbodiimide combined with ethanol-wet bonding pretreatment on dentin bonding properties: an in vitro study.

Purpose: This study evaluated the combined effects of Carbodiimide (EDC) and ethanol-wet bonding (EWB) pretreatment on the bond strength and resin-dentin surface. Methods: Phosphoric acid-etched dentin specimens were randomly divided into five groups based on the following pretreatments: deionized water (control), EWB, 0.3M EDC in water (EDCw), EDC water solution combined EWB (EDCw + EWB), and 0.3M EDC in ethanol (EDCe). A scanning electron microscope (SEM) was used to observe the morphology of collagen fibrils on the demineralized dentin matrix in each group after pretreatment. The adhesives Prime & Bond NT (PB) (Dentsply De trey, Konstanz, Germany) or Single bond 2 (SB) (3M ESPE, St. Paul, MN, USA) was applied after pretreatments, and a confocal laser scanning microscope (CLSM) was used to evaluate the quality of resin tags. The degree of conversion (DC) of the adhesive was investigated by Fourier transform infrared spectroscopy (ATR-FTIR). The dentin was first bonded with resin and bathed in water at 37 °C for 24 h. Half of them were subjected to 10, 000 cycles in a thermocycler between 5 °C and 55 °C before a microshear bond strength (µSBS) test. The statistical methods were Analysis of Variance (ANOVA) and a Tukey post hoc test at α = 0.05. Results: The µSBS was significantly affected by pretreatments (p < 0.001), adhesives (p < 0.001), and aging conditions (p < 0.001) as revealed by the three-way ANOVA. The EDCw, EDCw + EWB, and EDCe groups significantly increased the µSBS; the EDCw + EWB and EDCe groups produced the highest µSBS. In the EDC-containing groups, the SEM showed at the collagen fibrils in the dentin matrix formed a three-dimensional network structure in the tubules after cross-linking into sheets, and the hybrid layer formed thicker resin tags under a CLSM. In the EDC-containing groups, the CLSM observed an increase in the length of resin tags. PB showed a higher DC and bonding strength than SB, and the five pretreatment groups tested did not affect the DC of the two adhesives. Conclusions: In etch-and-rinse bonding system, EDC combined with EWB pretreatment can improve the quality of the hybrid layer and enhance the mechanical properties of demineralized dentin matrix. Pretreatment with EDC-ethanol solution may be a new clinically friendly option for enhancing dentin bonding durability.

A Novel Enzyme-Based SPR Strategy for Detection of the Antimicrobial Agent Chlorophene.

Chlorophene is an important antimicrobial agent present in disinfectant products which has been related to health and environmental effects, and its detection has been limited to chromatographic techniques. Thus, there is a lack of research that attempts to develop new analytical tools, such as biosensors, that address the detection of this emerging pollutant. Therefore, a new biosensor for the direct detection of chlorophene in real water is presented, based on surface plasmon resonance (SPR) and using a laccase enzyme as a recognition element. The biosensor chip was obtained by covalent immobilization of the laccase on a gold-coated surface through carbodiimide esters. The analytical parameters accomplished resulted in a limit of detection and quantification of 0.33 mg/L and 1.10 mg/L, respectively, fulfilling the concentrations that have already been detected in environmental samples. During the natural river's measurements, no significant matrix effects were observed, obtaining a recovery percentage of 109.21% ± 7.08, which suggested that the method was suitable for the fast and straightforward analysis of this contaminant. Finally, the SPR measurements were validated with an HPLC method, which demonstrated no significant difference in terms of precision and accuracy, leading to the conclusion that the biosensor reflects its potential as an alternative analytical tool for the monitoring of chlorophene in aquatic environments.

An empirical model for the evaluation of the dissolution rate from a DNAPL-contaminated area.

This paper investigates dynamic variation in the morphologic distribution of dense non-aqueous phase liquids (DNAPLs), which take into account the coupled mass transfer. Experiments were carried out in a 2D tank representing a reconstructed aquifer model. DNAPL dissolution rates were investigated over a wide range of DNAPL saturations, several source configurations, and different hydraulic conditions. Morphometric indexes are presented that take into consideration further factors affecting the dissolution process. Local information regarding transport parameters related to the characteristics of the medium was obtained through a neural network and an optimization algorithm applied to experimental tracer tests. The history of DNAPL source architecture, in terms of saturation, indentation grade, and orientation, was determined by image analysis. Dissolved concentrations were registered and mass transfer rate coefficients were obtained for a wide range of source-zone configurations. A statistical analysis was performed to develop a constitutive equation that is descriptive of the mass transfer rate as a function of source-zone metric characteristics. A new empirical dissolution model using the proposed morphometric parameters is presented and compared with other models. The mass transfer correlation reported incorporates morphometric parameters and considers the complex and variable architecture of non-miscible contaminants. The proposed correlation can be used for an initial assessment of non-aqueous phase liquid (NAPL) dissolution rates over a wide range of saturation (residual and non-residual) conditions and different aqueous phase velocities within the NAPL source zone.

Synthesis of esters of ginsenoside metabolite M1 and their cytotoxicity on MGC80-3 cells.

Monoesters of ginsenoside metabolite M1 at the 3-OH, 4-OH and 6-OH positions of the glucose moiety at M1 were synthesized via the reaction of M1 with acyl chloride, or acid-N,N'-diisopropylcarbodiimide in the presence of DMAP. Their structures were fully characterized by spectral methods. The cytotoxicity of these compounds against then MGC80-3 human gastric cancer cell line was also assessed. High inhibitory effects were found at a concentration of 100 µg/mL.

[Reaction of carbodiimide for the introduction of p-bis(2-chloroethyl)amino-L-phenylalanine to lysozyme]. / Réaction à la carbodiimide visant l'introduction de la p-bis-(2-chloroéthyl)amino-L-phénylalanine sur le lysozyme.

In the synthesis of new prodrugs with inhibitoring action of tumour growth, a new nitrogen mustard derivative was obtained, proceeding of the coupling between an egg-white lysozyme with an antitumor amine nucleophile, the methyl ester of p-bis-(2-chloroethyl)amino-L-phenylalanine (Melphalan), catalyzed by 1-ethyl-3-[3-(dimethylamino)propyl] carbodiimide (EDC), at pH 5.0 and room temperature. In that case, the mechanism for the modification isn't selective of Asp101 in lysozyme. As in cases of histamine and D-glucosamine [3], it is evident that Melphalan is one type of amine who doesn't cause a selective modification of Asp101 but causes somewhat random reaction, because Asp101 is modified followed by modifications of other carboxyls. In this case, we suggest that the amine (Melphalan) may also bind to the substrate binding site in competition with EDC. With this type of amine, enzyme-nucleophile interactions predominate, and the selective activation of Asp101 by EDC is reduced to lead a more random reaction.

Affinity precipitation of proteins using bis-dyes.

Based on the principle of affinity precipitation of nucleotide-dependent enzymes using bis-NAD (Mosbach), Lowe et al. have recently mentioned the possibility of synthesising bis-dyes for similar applications. In this paper we report preliminary results obtained using bis-dyes in the sulphonamide form got through carbodiimide condensation of the monomer and its aminohexyl derivative for affinity precipitation. The dimer exhibited considerable selectivity for lactate dehydrogenase (90% yield). Bovine serum albumin gave a lower yield of 50% and as expected chymosin could not be precipitated by the dimer.

Analytical, toxicological and immunological consequences of the use of N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide as coupling reagent for the preparation of meningococcal group C polysaccharide-tetanus toxoid conjugate as vaccine for human use.

For the preparation of meningococcal group C polysaccharide-tetanus toxoid conjugate the reactive reagent N-ethyl-N'-(dimethylaminopropyl) carbodiimide is used. The application of this reagent results in a number of stable linkages (viz. "peptide" linkages between the polysaccharide and tetanus toxoid, intrachain ester linkages in the polysaccharide component and binding of the N-acylurea derivative of the reagent) and less stable ones (viz. anhydride linkages). As a consequence of the reaction, the reagent is converted to a non-reactive urea derivative. The toxic properties of the reagent and of the converted reagent were studied. These properties do not contraindicate the use of the coupling reagent for the preparation of vaccines for human use. In addition analytical methods have been developed for the quantitative evaluation of the coupling reagent, the reaction products and for the N-acylurea derivative of the reagent and of the residual reactivity of conjugates for primary aminogroups. Although no test was performed for the assay of ester linkages in the polysaccharide component of the conjugate, evidence is presented that such linkages may be present. The results of the test for residual reactivity indicated a spontaneous rearrangement of linkages after the preparation of the conjugate. In addition the influence of the ratio of coupling reagent-to-polysaccharide and tetanus toxoid on antigenic and immunogenic activities of the conjugate was studied. An increase of the ratio resulted in a decrease of the antigenic activity of the polysaccharide component but in an increase of its immunogenic activity as to the induction of IgG antibodies to the polysaccharide. The immunogenic activity of the polysaccharide component correlated rather well with the antigenic activity measured in heterologous enzyme-linked immunosorbent assay using antibodies to both components.

Fluorometric determination of carbodiimides with trans-aconitic acid.

A new fluorometric method for the determination of carbodiimides with trans-aconitic acid as a reagent has been developed. With the present method, as little as 50 pmol of carbodiimides can be determined. The relative standard deviations obtained with 25 nmol of dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide HCl (EDC) were 1.35, 1.41, and 2.72% (n = 10), respectively.