Anesthesia of fish with benzocaine does not interfere with comet assay results

Fish blood erythrocytes are frequently used as sentinels in biomonitoring studies. Usually, fish blood is collected by painful cardiac or caudal vein punctures. Previous anesthesia could decrease animal suffering but it is not known at present whether anesthesia can cause confounding effects. Therefore, using the alkaline single cell gel (SCG)/comet assay with blood erythrocytes of the cichlid fish Nile tilapia, we tested for a possible modulation of induced DNA damage (methyl methanesulfonate; MMS) by the anesthetic benzocaine administered by bath exposure (80mg/l for ∼10min). Furthermore, benzocaine (80-600mg/l) was tested for its genotoxic potential on fish erythrocytes in vitro and for potential interactions with two known genotoxins (MMS and hydrogen peroxide). Our results did neither indicate a significant increase in the amount of DNA damage (even after a 48h follow-up), nor indicated interactions with MMS-induced DNA damage when fish were exposed to benzocaine in vivo. There was also no increase in DNA damage after in vitro exposure of fish erythrocytes to benzocaine. Clear concentration-related effects were observed for the two genotoxins in vitro, which were not significantly altered by the presence of benzocaine. These results suggest that anesthesia of fish does not confound comet assay results and the use of blood samples from anesthetized fish can be recommended with regard to animal welfare. © 2002 Elsevier Science B.V. All rights reserved.

Screening of Formulation Variables for the Preparation of Poly(epsilon-caprolactone) Nanocapsules Containing the Local Anesthetic Benzocaine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

pH mediated delamination of anionic clay-like nickel-zinc hydroxysalt in water through intercalation of zwitterionic p-aminobenzoate ions

p-aminobenzoate could be intercalated into the anionic clay, Ni3Zn2(OH)(8)(OAc)(2)center dot 2H(2)O at a high pH (similar to 10). When the pH was reduced to similar to 7 while washing colloidal dispersion due to delamination was observed. The development of partial positive charge on the amine end of the intercalated anion causes repulsion between the layers leading to delamination and colloidal dispersion of monolayers in water. The layers could be restacked from the colloid to form the parent solid either by increasing the pH or by evaporation.

p-Aminobenzoate ion determination in pharmaceutical formulations by using a potentiometric sensor immobilized in a graphite matrix

The characteristics, performance, and application of an electrode, namely, Pt\Hg\Hg-2(PABzt)(2)\ graphite, where PABzt stands for p-aminobenzoate ion, are described. This electrode responds to PABzt with sensivity of (58.1 +/- 1.0) mV per decade over the range 1.0 x 10(-4) to 1.0 x 10(-1) mol l(-1) at pH 6.5-8.0 and a detection limit of 3.2 x 10(-5) mol l(-1). The electrode shows easy construction, fast response time (within 10-30 s), low-cost, and excellent response stability (lifetime greater than 6 months, in continuous use). The proposed sensor displayed good selectivity for p-aminobenzoate in the presence of several substances, especially, concerning carboxylate and inorganic anions. It was used to determine p-aminobenzoate in pharmaceutical formulations by means of the standard additions method. The results obtained by using this electrode compared very favorably with those given by an HPLC procedure. (C) 2004 Elsevier B.V. All rights reserved.

Benzocaine loaded biodegradable poly-(D,L-lactide-co-glycolide) nanocapsules: factorial design and characterization

Local anesthetics are able to induce pain relief since they bind to the sodium channel of excitable membranes, blocking the influx of sodium ions and the propagation of the nervous impulse. Benzocaine (BZC) is a local anesthetic that presents limited application in topical formulations due to its low water-solubility. This study aimed to develop polymeric nanocapsules as a drug delivery system for the local anesthetic benzocaine (BZC). To do so, BZC loaded poly(D,L-lactide-co-glycolide) (PLGA) nanocapsules were prepared using the nanoprecipitation method and were characterized. The factorial experimental design was used to study the influence of four different independent variables oil response to nanocapsules drug loading. The physical characteristics of PLGA nanocapsules were evaluated by analyzing the particle size, the polydispersion index and the zeta potential, using a particle size analyzer. The results of the optimized formulation showed a size distribution with a polydispersity index of 0.12. an average diameter of 123 nm, zeta potential of -33.6 mV and a drug loading of more than 69%. The release profiles showed a significant difference in the release behavior for the pure drug in solution when compared with that containing benzocaine loaded PLGA nanocapsules. Thus, the prepared nonocapsules described here may be of clinical importance in both the processes of stabilization and delivery of benzocaine for pain treatment. (c) 2009 Elsevier B.V. All rights reserved.

Analysis of aromatic amines by high-performance liquid chromatography with pre-column derivatization by 2-(9-carbazole)-ethyl-chloroformate

2-(9-Carbazole)-ethyl-chloroformate (CEOC), a novel pre-column fluorescence derivatization reagent, has been developed for the analysis of aromatic amines. Taking five monocyclic aromatic amines (o-toluidine, aniline, 3,4-dimethylaniline, N-ethyl-p-toluidine, and p-phenylenediamine) as testing compounds, derivatization conditions such as pH of borate buffer, reaction time and fluorescent tagging reagent concentration have been investigated. By a one-step procedure, CEOC reacts readily with the aromatic amines to form stable derivatives with excitation and emission wavelengths, respectively, at 293 and 360 nm. This derivatization reaction could be finished within 20 min even at room temperature. The peak shapes of the derivatized aromatic amines can be improved greatly without any addition of competition amines into the mobile phase. Furthermore, this method can offer excellent quantitative precision with high tolerance of the matrix of samples. (C) 2003 Elsevier B.V. All rights reserved.

Desenvolvimento e caracterização de nanocápsulas de poli (L-lactídeo) contendo benzocaína

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Determinação de parâmetros físico-químicos com um sensor potenciométrico sensível ao íon p-aminobenzoato

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

MS222 does not induce primary DNA damage in fish

The genotoxicity of the anaesthetic MS222 (tricaine) was analysed in fish under both in vivo and in vitro conditions. Based on results of the single cell gel/Comet assay, MS222 had no direct genotoxic effect on the experimental fish, indicating that MS222 does not induce primary DNA damage. These results suggest that the use of this important anaesthetic in aquaculture can be considered to be safe in terms of genotoxicity. © Springer Science+Business Media, Inc. 2007.

Desenvolvimento e caracterização de nanocápsulas de poli(l-lactídeo) contendo benzocaína

In this paper we describe the preparation poly (L-lactide) (PLA) nanocapsules as a drug delivery system for the local anesthetic benzocaine. The characterization and in vitro release properties of the system were investigated. The characterization results showed a polydispersity index of 0.14, an average diameter of 190.1± 3 nm, zeta potential of –38.5 mV and an entrapment efficiency of 73%. The release profile of Benzocaine loaded in PLA nanocapsules showed a significant different behavior than that of the pure anesthetic in solution. This study is important to characterize a drug release system using benzocaine for application in pain treatment.

Desenvolvimento e caracterização de nanocápsulas de poli (L-lactídeo) contendo benzocaína

In this paper we describe the preparation poly (L-lactide) (PLA) nanocapsules as a drug delivery system for the local anesthetic benzocaine. The characterization and in vitro release properties of the system were investigated. The characterization results showed a polydispersity index of 0.14, an average diameter of 190.1± 3 nm, zeta potential of -38.5 mV and an entrapment efficiency of 73%. The release profile of Benzocaine loaded in PLA nanocapsules showed a significant different behavior than that of the pure anesthetic in solution. This study is important to characterize a drug release system using benzocaine for application in pain treatment.

Purification and properties of benzoate-4-hydroxylase from a soil pseudomonad

Benzoate-4-hydroxylase from a soil pseudomonad was isolated and purified about 50-fold. Polyacrylamide gel electrophoresis of this enzyme preparation showed one major band and one minor band. The approximate molecular weight of the enzyme was found to be 120,000. Benzoate-4-hydroxylase was most active around pH 7.2. The enzyme showed requirements for tetrahydropteridine as the cofactor and molecular oxygen as the electron acceptor. NADPH, NADH, dithiothreitol, β-mercaptoethanol, and ascorbic acid when added alone to the reaction mixture did not support the hydroxylation reaction to any significant extent. However, when these compounds were added together with tetrahydropteridine, they stimulated the hydroxylation. This stimulation is probably due to the reduction of the oxidized pteridine back to the reduced form. This enzyme was activated by Fe2+ and benzoate. It was observed that benzoate-4-hydroxylase could catalyze the oxidation of NADPH in the presence of benzoate,p-aminobenzoate, p-nitrobenzoate, p-chlorobenzoate, and p-methylbenzoate, with only benzoate showing maximum hydroxylation. Inhibition studies with substrate analogs and their kinetic analysis revealed that the carboxyl group is involved in binding the substrate to the enzyme at the active center. The enzyme catalyzed the conversion of 1 mol of benzoate to 1 mol of p-hydroxybenzoate with the consumption of slightly more than 1 mol of NADPH and oxygen.

Effect of clofibrate on the adenosine triphosphatase activity of rat liver mitochondria

The antihypercholesterolemic drug clofibrate (ethyl-α-p-chlorophenoxyisobutyrate) stimulated the latent ATPase activity and “superstimulated” the uncoupler-induced ATPase activity of rat-liver mitochondria. Addition of clofibrate decreased the turbidity of mitochondrial suspensions and released considerable amount of mitochondrial protein into solution. In these properties it closely resembled detergents like Triton X-100 and deoxycholate. However, unlike the detergents, clofibrate required the presence of a permeant cation for its disruptive action. Also, it was without any such effect on sonic submitochondrial particles. The drug enhanced the uptake of both Mg2 and Cl− by mitochondria suggesting that osmotic swelling precedes lysis. Sonic submitochondrial particles prepared in the presence of clofibrate showed a greater yield and comparable ATPase activity.

Exfoliation of alpha-hydroxides of nickel and cobalt in water

p-Aminobenzoate ion intercalated alpha-hydroxides of nickel/cobalt were synthesized by precipitation using ammonia (pH = similar to 12). Aqueous colloidal suspension of alpha-hydroxide of nickel/cobalt was obtained on washing the precipitate as the pH was reduced to similar to 7. The development of partial positive charge on the amine end of the intercalated anion causes repulsion between the layers leading to exfoliation and colloidal suspension of monolayers in water. While theb layers could be restacked from the colloidal suspension in the presenceof other anions in the case of alpha-cobalt hydroxide, the exfoliation could not be reversed easily in the case of the nickel analog. (C) 2010 Elsevier Inc. All rights reserved.

A series of transition metal-azido extended complexes with various anionic and neutral co-ligands: synthesis, structure and their distinct magnetic behavior

The crystal structures and magnetic properties of five new transition metal-azido complexes with two anionic [pyrazine-2-carboxylate (pyzc) and p-aminobenzoate (paba)] and two neutral [pyrazine (pyz) and pyridine (py)] coligands are reported All five complexes were synthesized bysolvothermal methods The complex [Co-2(pyzc)(2)(N-3)(2)(H2O)(2)](n) (1) is 1D and exhibit canted antiferromagnetism, while the 3D complex [MnNa(pyzc)(N-3)(2)(H2O)(2)](n) (2) has a complicated structure and is weakly ferromagnetic in nature [Mn-2(paba)(2)(N-3)(2)(H2O)(2)](n) (3). is a 2D sheet and the Mn-II ions are found to be antiferromagnetically coupled The isostructural 2D complexes [Cu-3(pyz)(2)(N-3)(6)](n) (4) and [Cu-3(py)(2)(N-3)(6)](n) (5) resemble remarkably in their magnetic properties exhibiting moderately strong ferromagnetism. Density functional theory calculations (B3LYP functional) have been performed to provide a qualitative theoretical interpietation of the overall magnetic behavior shown by these complexes.