Easy introduction of maleimides at different positions of oligonucleotide chains for conjugation purposes

[2,5-Dimethylfuran]-protected maleimides were placed at both internal positions and the 3'-end of oligonucleotides making use of solid-phase synthesis procedures. A new phosphoramidite derivative and a new solid support incorporating the protected maleimide moiety were prepared for this purpose. In all cases maleimide deprotection (retro-Diels-Alder reaction) followed by reaction with thiol-containing compounds afforded the target conjugate.

Ação e caracterização química de óleos essenciais no manejo da antracnose do maracujá

Objetivou-se avaliar dois métodos de inoculação de Colletotrichum gloeosporioides em maracujá, testar a patogenicidade de diferentes isolados, o efeito fungitóxico e a composição química dos óleos essenciais das espécies medicinais alecrim-pimenta (Lippia sidoides Cham.), capim-santo [Cymbopogon citratus (D. C.) Stapf.], alfavaca-cravo (Ocimum gratissimum L.), no controle da antracnose [Colletotrichum gloeosporioides (Penz.)], associado ao estádio de maturação de frutos de maracujazeiro-amarelo. Avaliaram-se três experimentos, onde se testou a patogenicidade de seis isolados do fungo em delineamento inteiramente casualizado, com seis repetições, outro com o mesmo delineamento em esquema fatorial 2x2 (suspensão de conídios e disco de micélio) e frutos (verdes e maduros), com seis repetições. No tratamento com frutos, utilizou-se o delineamento inteiramente casualizado, em esquema fatorial 5x3+1, sendo cinco concentrações (0; 2; 4; 6 e 8µL mL-1) e três espécies medicinais, mais o tebuconazol, com cinco repetições. Fez-se a caracterização química dos óleos por cromatografia gasosa, com espectrometria de massas. Todos os isolados foram patogênicos. Os frutos maduros apresentaram maior diâmetro das lesões, quando inoculados com suspensão de conídios. O óleo de C. citratus proporcionou o menor diâmetro das lesões nos frutos, até a concentração de 6 µL mL-1. Na concentração de 8 µL mL-1, todos os óleos inibiram o desenvolvimento do fungo. O timol (30,24%), o citral (77,74%) e o eugenol (92,89%) foram componentes majoritários em L. sidoides, C. citratus e O. gratissimum, respectivamente.

Microscopic-macroscopic approach for binding energies with the Wigner-Kirkwood method

The semiclassical Wigner-Kirkwood ̄h expansion method is used to calculate shell corrections for spherical and deformed nuclei. The expansion is carried out up to fourth order in ̄h. A systematic study of Wigner-Kirkwood averaged energies is presented as a function of the deformation degrees of freedom. The shell corrections, along with the pairing energies obtained by using the Lipkin-Nogami scheme, are used in the microscopic-macroscopic approach to calculate binding energies. The macroscopic part is obtained from a liquid drop formula with six adjustable parameters. Considering a set of 367 spherical nuclei, the liquid drop parameters are adjusted to reproduce the experimental binding energies, which yields a root mean square (rms) deviation of 630 keV. It is shown that the proposed approach is indeed promising for the prediction of nuclear masses.

Synthesis of Janus compounds for the recognition of G-U mismatched nucleobase pairs

The design and synthesis of two Janus-type heterocycles with the capacity to simultaneously recognize guanine and uracyl in G-U mismatched pairs through complementary hydrogen bond pairing is described. Both compounds were conveniently functionalized with a carboxylic function and efficiently attached to a tripeptide sequence by using solid-phase methodologies. Ligands based on the derivatization of such Janus compounds with a small aminoglycoside, neamine, and its guanidinylated analogue have been synthesized, and their interaction with Tau RNA has been investigated by using several biophysical techniques, including UV-monitored melting curves, fluorescence titration experiments, and 1H NMR. The overall results indicated that Janus-neamine/guanidinoneamine showed some preference for the +3 mutated RNA sequence associated with the development of some tauopathies, although preliminary NMR studies have not confirmed binding to G-U pairs. Moreover, a good correlation has been found between the RNA binding affinity of such Janus-containing ligands and their ability to stabilize this secondary structure upon complexation.

Redução de incertezas em radioterapia utilizando simulação Monte Carlo: análise espectral aplicada à correção de dose absorvida

OBJETIVO: Determinar, por simulação Monte Carlo, os espectros de feixes de cobaltoterapia em profundidade na água e fatores de correção para doses absorvidas em dosímetros termoluminescentes de fluoreto de lítio. MATERIAIS E MÉTODOS: As simulações dos espectros secundários da fonte clínica de cobalto-60 foram realizadas com o código Monte Carlo PENELOPE, em diversas profundidades na água. Medidas experimentais de dose profunda foram obtidas com dosímetros termoluminescentes e câmara de ionização em condições de referência em radioterapia. Os fatores de correção para os dosímetros termoluminescentes foram obtidos através da razão entre as absorções relativas ao espectro de baixa energia e ao espectro total. RESULTADOS: A análise espectral em profundidade revelou a existência de espectros secundários de baixa energia responsáveis por uma parcela significativa da deposição de dose. Foram observadas discrepâncias de 3,2% nas doses medidas experimentalmente com a câmara de ionização e com os dosímetros termoluminescentes. O uso dos fatores de correção nessas medidas permitiu diminuir a discrepância entre as doses absorvidas para, no máximo, 0,3%. CONCLUSÃO: Os espectros simulados permitem o cálculo de fatores de correção para as leituras de dosímetros termoluminescentes utilizados em medidas de dose profunda, contribuindo para a redução das incertezas associadas ao controle de qualidade de feixes clínicos em radioterapia.

Identification of new transformation products during enzymatic treatment of tetracycline and erythromycin antibiotics at laboratory scale by an on-line turbulent flow liquid-chromatography coupled to a high resolution mass spectrometer LTQ-Orbitrap

This work describes the formation of transformation products (TPs) by the enzymatic degradation at laboratory scale of two highly consumed antibiotics: tetracycline (Tc) and erythromycin (ERY). The analysis of the samples was carried out by a fast and simple method based on the novel configuration of the on-line turbulent flow system coupled to a hybrid linear ion trap – high resolution mass spectrometer. The method was optimized and validated for the complete analysis of ERY, Tc and their transformation products within 10 min without any other sample manipulation. Furthermore, the applicability of the on-line procedure was evaluated for 25 additional antibiotics, covering a wide range of chemical classes in different environmental waters with satisfactory quality parameters. Degradation rates obtained for Tc by laccase enzyme and ERY by EreB esterase enzyme without the presence of mediators were ∼78% and ∼50%, respectively. Concerning the identification of TPs, three suspected compounds for Tc and five of ERY have been proposed. In the case of Tc, the tentative molecular formulas with errors mass within 2 ppm have been based on the hypothesis of dehydroxylation, (bi)demethylation and oxidation of the rings A and C as major reactions. In contrast, the major TP detected for ERY has been identified as the “dehydration ERY-A”, with the same molecular formula of its parent compound. In addition, the evaluation of the antibiotic activity of the samples along the enzymatic treatments showed a decrease around 100% in both cases

Identification of metabolites and thermal transformation products of quinolones in raw cow milk by liquid chromatography coupled to high resolution mass spectrometry

The presence of residues of antibiotics, metabolites, and thermal transformation products (TPs), produced during thermal treatment to eliminate pathogenic microorganisms in milk, could represent a risk for people. Cow"s milk samples spiked with enrofloxacin (ENR), ciprofloxacin (CIP), difloxacin (DIF), and sarafloxacin (SAR) and milk samples from cows medicated with ENR were submitted to several thermal treatments. The milk samples were analyzed by liquid chromatography-mass spectrometry (LC-MS) to find and identify TPs and metabolites. In this work, 27 TPs of 4 quinolones and 24 metabolites of ENR were found. Some of these compounds had been reported previously, but others were characterized for the first time, including lactose-conjugated CIP, the formamidation reaction for CIP and SAR, and hydroxylation or ketone formation to produce three different isomers for all quinolones studied.

Comparison of three analytical methods for the determination of quinolones in pig muscle samples

This work presents a comparison between three analytical methods developed for the simultaneous determination of eight quinolones regulated by the European Union (marbofloxacin, ciprofloxacin, danofloxacin, enrofloxacin, difloxacin, sarafloxacin, oxolinic acid and flumequine) in pig muscle, using liquid chromatography with fluorescence detection (LC-FD), liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The procedures involve an extraction of the quinolones from the tissues, a step for clean-up and preconcentration of the analytes by solid-phase extraction and a subsequent liquid chromatographic analysis. The limits of detection of the methods ranged from 0.1 to 2.1 ng g−1 using LC-FD, from 0.3 to 1.8 using LC-MS and from 0.2 to 0.3 using LC-MS/MS, while inter- and intra-day variability was under 15 % in all cases. Most of those data are notably lower than the maximum residue limits established by the European Union for quinolones in pig tissues. The methods have been applied for the determination of quinolones in six different commercial pig muscle samples purchased in different supermarkets located in the city of Granada (south-east Spain).

Identification of metabolites and thermal transformation products of quinolones in raw cow milk by liquid chromatography coupled to high resolution mass spectrometry.

The presence of residues of antibiotics, metabolites, and thermal transformation products (TPs), produced during thermal treatment to eliminate pathogenic microorganisms in milk, could represent a risk for people. Cow"s milk samples spiked with enrofloxacin (ENR), ciprofloxacin (CIP), difloxacin (DIF), and sarafloxacin (SAR) and milk samples from cows medicated with ENR were submitted to several thermal treatments. The milk samples were analyzed by liquid chromatography-mass spectrometry (LC-MS) to find and identify TPs and metabolites. In this work, 27 TPs of 4 quinolones and 24 metabolites of ENR were found. Some of these compounds had been reported previously, but others were characterized for the first time, including lactose-conjugated CIP, the formamidation reaction for CIP and SAR, and hydroxylation or ketone formation to produce three different isomers for all quinolones studied.

Obtención de láminas delgadas de Y-Ba-Cu-O sobre Si (100)

Se ha utilizado la evaporación secuencial de Cu, Y2O3 y BaF2 para la obtención de láminas delgadas superconductoras de YBa-CuO sobre substratos de Si monocristalino con orientación [100], recubiertos con una lámina barrera de Zr02. Se han estudiado los efectos de la variación de los espesores relativos de las láminas constituyentes y del espesor total de la lámina resultante. Las láminas se han caracterizado mediante medidas de la variación de la resistencia con la temperatura, microscopía electrónoca de barrido, difractometría de rayos X, microson-da electrónica y espectometría de masas de iones secundarios. Las láminas presentan un ligero carácter semiconductor en el estado normal, con temperaturas de inicio de la transición su-perconductora alrededor de 90 K, y resistencia nula, en el mejor de los casos, a 45 K.

Nutrimetabolomics fingerprinting to identify biomarkers of bread exposure in a free-living population from the PREDIMED study cohort

Bread is one of the most widely consumed foods. Its impact on human health is currently of special interest for researchers. We aimed to identify biomarkers of bread consumption by applying a nutrimetabolomic approach to a free-living population. An untargeted HPLC q-TOF-MS and multivariate analysis was applied to human urine from 155 subjects stratified by habitual bread consumption in three groups: non-consumers of bread (n = 56), white-bread consumers (n = 48) and whole-grain bread consumers (n = 51). The most differential metabolites (variable importance for projection ≥1.5) included compounds originating from cereal plant phytochemicals such as benzoxazinoids and alkylresorcinol metabolites, and compounds produced by gut microbiota (such as enterolactones, hydroxybenzoic and dihydroferulic acid metabolites). Pyrraline, riboflavin, 3-indolecarboxylic acid glucuronide, 2,8-dihydroxyquinoline glucuronide and N-α-acetylcitrulline were also tentatively identified. In order to combine multiple metabolites in a model to predict bread consumption, a stepwise logistic regression analysis was used. Receiver operating curves were constructed to evaluate the global performance of individual metabolites and their combination. The area under the curve values [AUC (95 % CI)] of combined models ranged from 77.8 % (69.1 86.4 %) to 93.7 % (89.4 98.1 %), whereas the AUC for the metabolites included in the models had weak values when they were evaluated individually: from 58.1 % (46.6 69.7 %) to 78.4 % (69.8 87.1 %). Our study showed that a daily bread intake significantly impacted on the urinary metabolome, despite being examined under uncontrolled free-living conditions. We further concluded that a combination of several biomarkers of exposure is better than a single biomarker for the predictive ability of discriminative analysis.

Nutrimetabolomics fingerprinting to identify biomarkers of bread exposure in a free-living population from the PREDIMED study cohort

Bread is one of the most widely consumed foods. Its impact on human health is currently of special interest for researchers. We aimed to identify biomarkers of bread consumption by applying a nutrimetabolomic approach to a free-living population. An untargeted HPLC q-TOF-MS and multivariate analysis was applied to human urine from 155 subjects stratified by habitual bread consumption in three groups: non-consumers of bread (n = 56), white-bread consumers (n = 48) and whole-grain bread consumers (n = 51). The most differential metabolites (variable importance for projection ≥1.5) included compounds originating from cereal plant phytochemicals such as benzoxazinoids and alkylresorcinol metabolites, and compounds produced by gut microbiota (such as enterolactones, hydroxybenzoic and dihydroferulic acid metabolites). Pyrraline, riboflavin, 3-indolecarboxylic acid glucuronide, 2,8-dihydroxyquinoline glucuronide and N-α-acetylcitrulline were also tentatively identified. In order to combine multiple metabolites in a model to predict bread consumption, a stepwise logistic regression analysis was used. Receiver operating curves were constructed to evaluate the global performance of individual metabolites and their combination. The area under the curve values [AUC (95 % CI)] of combined models ranged from 77.8 % (69.1 86.4 %) to 93.7 % (89.4 98.1 %), whereas the AUC for the metabolites included in the models had weak values when they were evaluated individually: from 58.1 % (46.6 69.7 %) to 78.4 % (69.8 87.1 %). Our study showed that a daily bread intake significantly impacted on the urinary metabolome, despite being examined under uncontrolled free-living conditions. We further concluded that a combination of several biomarkers of exposure is better than a single biomarker for the predictive ability of discriminative analysis.

Potencialidades e aplicações do ligante dietilditiofosfato de amônio em análises químicas

Este trabalho descreve uma revisão a respeito do uso do ligante dietilditiofosfato de amônio em química analítica e suas potencialidades no campo da análise química espectrofotométrica (UV/VIS) e espectrométrica (AAS, ICP-AES, ICP-MS) em diferentes matrizes e meios. A discriminação de espécies químicas por meio de complexações seletivas de elementos químicos com dietilditiofosfato e a hifenação de técnicas espectroscópicas a etapas de separação como extração em fase sólida, extração líquido-líquido, extração por "cloud point", são outros assuntos abordados neste trabalho.

Análise química inorgânica de leite esimulação do reagente CFA-C

Na análise de leite por espectrometria de emissão ótica com plasma induzido (ICP-OES) o produto comercial CFA-C, constituído por aminas terciárias, vêm sendo usado com sucesso como diluente em fase única. Devido ao custo do produto e possibilidades de contaminação durante o armazenamento, decidiu-se desenvolver um outro reagente de propriedades equivalentes ao referido produto. Testes qualitativos mostraram que o produto não contém ácido bórico nem EDTA. Em uma titulação potenciométrica de uma solução aquosa 10% v/v CFA-C com HCl obtiveram-se dois picos potenciométricos e deduziu-se que 90% do ácido foi consumido para neutralização da amina livre e 10% foi gasto para protonar um ânion de ácido fraco usado para formar um tampão de razão molar 10:1, do tipo R3N/R3NH+. Em uma simulação desse reagente preparou-se uma solução 6,06 mol L-1 de monoetanolamina (MEA) e 0,20 mol L-1 ácido cítrico, triprótico, para formar um tampão do mesmo tipo, 10:1, baseado na propriedade do íon citrato em complexar Ca(II). Outro tampão citrato mais diluído (1,52 mol L-1) foi preparado usando trishidroximetilaminometano (TRIS), que é uma amina menos solúvel. Essas misturas foram caracterizadas por análise potenciométrica e termoanalítica. O comportamento desses novos tampões amina/citrato foi adequado e comparável ao produto comercial.

Análise exploratória dos teores de constituintes inorgânicos em sucos e refrigerantes de uva

Os teores de Ca, Cu, K, Mg, Na, P e S foram determinados simultaneamente em sucos e refrigerantes de uva utilizando espectrometria de emissão ótica com plasma de argônio induzido (ICP-OES) como técnica analítica. Os dados obtidos foram submetidos a dois métodos de análise exploratória: análise de componentes principais (PCA) e análise hierárquica de agrupamentos (HCA). Os resultados mostraram que o Na foi o principal elemento para discriminação entre as amostras, diferenciando principalmente sucos e refrigerantes. A adição de conservantes e o teor de sais minerais também foram aspectos relevantes para a discriminação dos grupos. A análise exploratória mostrou-se útil para o tratamento dos dados em questão, produzindo informações que dificilmente seriam visualizadas diretamente na matriz de dados.