Hexokinase production fromS. cerevisiae

Applied Biochemistry and Biotechnology - The effects of pH (4.0, 4.5, or 5.0), temperature (T) (30, 35, or 40°C) and dissolved oxygen (DO) (0.2, 2.0, 4.0, or 6.0 mg O2/L) on hexokinase and...     

Screening of fluoroquinolones in environmental waters using disk-based solid-phase extraction combined to microplate fluorimetric determination and LC-MS/MS

Fluoroquinolones are in the order of the day concerning environmental contamination through anthropogenic activities, resulting in increased risk for antibiotic resistance dissemination. In this context, accessible, low-cost analytical methods are required for implementation of comprehensive surveillance and screening schemes. In this work, we propose a down-scaled disk-based solid-phase extraction system from which the eluate can be first screened by miniaturized fluorimetric reading, followed by individual determination of target fluoroquinolones (ciprofloxacin, norfloxacin, and enrofloxacin) by liquid chromatography combined to tandem mass spectrometry. The fluorimetric measurement is based on the intrinsic fluorescence of fluoroquinolones. Disk-based retention was performed after sample acidification (pH 4.0) by mixed-mode cation exchange using polystyrene divinylbenzene sulphonated sorbent. Sample loading was precisely controlled in a dedicated flow system operating at 4.0 mL min^?1. Different eluent compositions were tested, with elution performed by 1.00 mL of methanol-ammonium hydroxide (98:2, v/v), with subsequent reading of eluate in both detectors. Quantification was attained for 2–25 µg L^?1 range, with LOD values at 1 µg L^?1. The proposed approach was successfully applied to estuarine waters from the Douro River, with comparable results to a conventional SPE-LC-MS/MS procedure.     

Effect of low intensity helium-neon (HeNe) laser irradiation on experimental paracoccidioidomycotic wound healing dynamics

The effect of HeNe laser on the extracellular matrix deposition, chemokine expression and angiogenesis in experimental paracoccidioidomycotic lesions was investigated. At days 7, 8 and 9 postinfection the wound of each animal was treated with a 632.8 nm HeNe laser at a dose of 3 J cm(-2). At day 10 postinfection, the wounds were examined by using histologic and immunohistochemical methods. Results revealed that laser-treated lesions were lesser extensive than untreated ones, and composed mainly by macrophages and lymphocytes. High IL-1beta expression was shown in the untreated group whereas in laser-treated animals the expression was scarce. On the other hand, the expression of CXCL-10 was found to be reduced in untreated animals and quite intensive and well distributed in the laser-treated ones. Also, untreated lesions presented vascular endothelial growth factor (VEGF) in a small area near the center of the lesion and high immunoreactivity for hypoxia-inducible factor-1 (HIF-1), whereas laser-treated lesions expressed VEGF surrounding blood vessels and little immunoreactivity for HIF-1. Laser-treated lesions presented much more reticular fibers and collagen deposition when compared with the untreated lesion. Our results show that laser was efficient in minimizing the local effects observed in paracoccidioidomycosis and can be an efficient tool in the treatment of this infection, accelerating the healing process.     

Palladium-catalyzed Suzuki cross-coupling of 2-haloselenophenes: synthesis of 2-arylselenophenes, 2,5-diarylselenophenes, and 2-arylselenophenyl ketones

We present herein our results on the Suzuki coupling reaction of 2-haloselenophenes with boronic acids catalyzed by palladium salt and describe a new route established to prepare 2-arylselenophenes and 2,5-diarylselenophenes in good yields. The reaction proceeded cleanly under mild conditions and was performed with aryl boronic acids bearing electron-withdrawing, electron-donating, and neutral substituents, in the presence of Pd(OAc)2, K2CO3/H2O in DME. In addition, by this protocol unsymmetrical aryl ketones were also obtained from 2-iodoselenophene and boronic acids via a carbonylative process.     

A new minor dimmeric ester from seeds of Cassia fistula L. (Leguminosae)

Bioactivity-guided fractionation of n-hexane phase from MeOH extract of the seeds of Cassia fistula L. (Leguminosae) yielded two bioactive substances against Cladosporium cladosporioides and C. sphaerospermum. After spectroscopic analysis, these compounds were characterised as the known benzyl 2-hydroxy-3,6-dimethoxybenzoate and its dimer dibenzyl 2,2'-dihydroxy-3,6,3″,6″-tetramethoxy-biphenyl-1,1'-dicarboxylate, which showed a new structural arrangement.     

Crossover to the KPZ Equation

We characterize the crossover regime to the KPZ equation for a class of one-dimensional weakly asymmetric exclusion processes. The crossover depends on the strength asymmetry an ^2-γ (a, γ >  0) and it occurs at γ = 1/2. We show that the density field is a solution of an Ornstein-Uhlenbeck equation if \({\gamma\in(1/2,1]}\) , while for γ = 1/2 it is an energy solution of the KPZ equation. The corresponding crossover for the current of particles is readily obtained.     

Keto-Enolic Equilibria of an Isatin-Schiff Base Copper(II) Complex and its Reactivity toward Carbohydrate Oxidation

An interesting isatin-Schiff base copper(II) complex, [Cu(isapn)](ClO₄)₂ where isapn= N,N-[bis-(3,3-indolin-2-one)]-1,3-diiminepropane, was prepared and characterized by different techniques, both in the solid state and in solution, and its reactivity toward carbohydrate oxidation was verified. The positive ion electrospray mass spectrum detects the complex as an isotopologue cluster of singly charged intact isatin-copper(I) ions of m/z 395 (for ⁶⁵Cu) with an isotopic pattern identical to that calculated for C₁₉H₁₆CuN₄O₂ ⁺. Tandem mass spectrometry reveals an interesting and structurally diagnostic collision-induced dissociation behavior for this ionized complex, which is dominated by the cleavage of the N—(CH₂)₃—N propylene bridge. In aqueous solution, this complex undergoes a peculiar keto-enolic equilibrium, verified at different pH's by spectroscopic methods (u.v.–vis. and e.p.r.), with a corresponding pK _a determined as 9.5. The e.p.r. parameter ratio g /A for this complex, in frozen MeOH/H₂O (4:1, v/v) solution at 77 K, changes from 188 cm in acidic medium (pH 2.5–3.0) to 118 cm in basic medium (pH 11), indicating a significant structural change from a distorted tetrahedral to a more tetragonal geometry around the copper ion. This compound was shown to catalyze the oxidation of hexoses (glucose, fructose and galactose), in alkaline media, via reactive oxygen species, which were detected by using specific enzymes, and by e.p.r. spin trapping. The reaction was monitored at (25.0 ± 0.1)°C by the consumption of oxygen, and showed first-order dependence on catalyst, followed by a saturation effect. First-order kinetics with respect to [OH^–] concentration was also observed, indicating that enolization of the substrate as well as the metal-catalyzed enediol oxidation are the rate-determining steps.     

Analysis of phase transition and dehydration processes of nevirapine

Solid-state characterization of crystalline drugs is an important pre-formulation step for the development and design of solid dosage forms, such as pellets and tablets. In this study, phase transition and dehydration processes of nevirapine have been studied by differential scanning calorimetry and thermogravimetry differential thermal analysis to overcome the problems of drug formulation, namely poor solubility and poor content uniformity. Phase solubility studies elucidated the mechanism of enhanced nevirapine solubility.