The unsolved puzzle of neuropathogenesis in glutaric aciduria type I.

Glutaric aciduria type I (GA-I) is a cerebral organic aciduria caused by deficiency of glutaryl-Co-A dehydrogenase (GCDH). GCDH deficiency leads to accumulation of glutaric acid (GA) and 3-hydroxyglutaric acid (3-OHGA), two metabolites that are believed to be neurotoxic, in brain and body fluids. The disorder usually becomes clinically manifest during a catabolic state (e.g. intercurrent illness) with an acute encephalopathic crisis that results in striatal necrosis and in a permanent dystonic-dyskinetic movement disorder. The results of numerous in vitro and in vivo studies have pointed to three main mechanisms involved in the metabolite-mediated neuronal damage: excitotoxicity, impairment of energy metabolism and oxidative stress. There is evidence that during a metabolic crisis GA and its metabolites are produced endogenously in the CNS and accumulate because of limiting transport mechanisms across the blood-brain barrier. Despite extensive experimental work, the relative contribution of the proposed pathogenic mechanisms remains unclear and specific therapeutic approaches have yet to be developed. Here, we review the experimental evidence and try to delineate possible pathogenetic models and approaches for future studies.

Sublimation of new matrix candidates for high spatial resolution imaging mass spectrometry of lipids: enhanced information in both positive and negative polarities after 1,5-diaminonapthalene deposition.

Matrix sublimation has demonstrated to be a powerful approach for high-resolution matrix-assisted laser desorption ionization (MALDI) imaging of lipids, providing very homogeneous solvent-free deposition. This work presents a comprehensive study aiming to evaluate current and novel matrix candidates for high spatial resolution MALDI imaging mass spectrometry of lipids from tissue section after deposition by sublimation. For this purpose, 12 matrices including 2,5-dihydroxybenzoic acid (DHB), sinapinic acid (SA), α-cyano-4-hydroxycinnamic acid (CHCA), 2,6-dihydroxyacetphenone (DHA), 2',4',6'-trihydroxyacetophenone (THAP), 3-hydroxypicolinic acid (3-HPA), 1,8-bis(dimethylamino)naphthalene (DMAN), 1,8,9-anthracentriol (DIT), 1,5-diaminonapthalene (DAN), p-nitroaniline (NIT), 9-aminoacridine (9-AA), and 2-mercaptobenzothiazole (MBT) were investigated for lipid detection efficiency in both positive and negative ionization modes, matrix interferences, and stability under vacuum. For the most relevant matrices, ion maps of the different lipid species were obtained from tissue sections at high spatial resolution and the detected peaks were characterized by matrix-assisted laser desorption ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry. First proposed for imaging mass spectrometry (IMS) after sublimation, DAN has demonstrated to be of high efficiency providing rich lipid signatures in both positive and negative polarities with high vacuum stability and sub-20 μm resolution capacity. Ion images from adult mouse brain were generated with a 10 μm scanning resolution. Furthermore, ion images from adult mouse brain and whole-body fish tissue sections were also acquired in both polarity modes from the same tissue section at 100 μm spatial resolution. Sublimation of DAN represents an interesting approach to improve information with respect to currently employed matrices providing a deeper analysis of the lipidome by IMS.

Meat lipid profile of steers finished in pearl millet pasture with different rates of concentrate

The objective of this work was to evaluate the meat lipid profile from Devon beef steers finished in pearl millet (Pennisetum americanum) pasture and fed at different rates of concentrate supplementary diet. Twelve steers weighing 270 kg, at 12‑month‑average initial age, were randomly distributed into three treatments: pearl millet pasture; and pearl millet pasture plus a concentrate equivalent at 0.5 or 1.0% of body weight, with two replicates. Total contents of saturated and unsaturated fatty acids, the polyunsaturated:saturated ratio and other relevant fatty acids as the vaccenic acid, conjugated linoleic acid, omega‑3, and omega‑6 were not affected by the consumption of a concentrate supplement at 0.5 or 1.0% live weight. However, the 0.5% supplementation level reduced the concentration of dihomo‑γ‑linolenic fatty acid (C20: 3 n‑6), while the 1.0% supplementation level elevated the content of docosahexaenoic (DHA) (C22: 6 n‑3) fatty acid, and the omega‑6:omega‑3 ratio in meat. Consumption of up to 1.0% energy supplementation increases the omega‑6:omega‑3 ratio in meat from Devon steers grazing on pearl millet pasture.

Síntese do butirato de n-butila empregando lipase microbiana imobilizada em copolímero de estireno-divinilbenzeno

This work investigates the reaction parameters of an immobilized lipase in the esterification reaction of n-butanol and butyric acid. Microbial lipase from Candida rugosa was immobilized onto styrene-divinylbenzene copolymer (STY-DVB) and subsequently introduced in an organic medium containing substrates in appropriate concentrations. Heptane was selected as solvent on the basis of its compatibility with the resin and the enzyme. The influence of molar ratio of acid to alcohol, amount of immobilized lipase and temperature on the butyl butyrate formation was determined. The results were compared with those achieved with free lipase and Lipozyme (commercially immobilized lipase) under the same operational conditions.

Flavonóide e triterpenos de Stigmaphyllom paralias

Stigmaphyllom paralias is a herb belonging to the family Malpighiaceae that occurs in sand soil of brazilian " restinga". This is the first report regarding phytochemical study with this species. The hexane extract of the aerial parts of plant afforded the triterpenes friedelin, lupenone, 3-oxo-alpha-amirin and 3-oxo-beta-amirin, the mixture of alpha-amirinyl palmitate and stearate, lupeol and 3,4-seco-friedelan-3-oic acid. The AcOEt extract yielded the flavonoid luteolin-7-rutinoside. All compounds were characterized by analysis of spectrometric data and the fatty acids esterified with alpha-amirine were identified by GC/MS of methyl derivatives of transesterified products . This is the first natural occurence of 3,4-seco-friedelan-3-oic acid and the 13C NMR spectral data were inequivocally assigned by two-dimensional techniques. This work also permitted to correct the 13C NMR resonances attributed to methyl groups C-26 and C-27 of fridelin.

Constituintes químicos de Melaleuca alternifolia (Myrtaceae)

The first chemical study of non-volatile constituents from the bark and stem of Melaleuca alternifolia (Myrtaceae) led to the isolation and identification of 3,3'-O-dimethylellagic acid (1) and five pentacyclic triterpenes: 2alpha,3beta,23-trihydroxyolean-12-en-28-oic acid (arjunolic acid, 2), 3beta-hydroxylup-20(29)-en-27,28-dioic acid (melaleucic acid, 3), betulinic acid (4), betuline (5), 3beta-O-acetylurs-12-en-28-oic acid (6), a mixture of fatty acids and esters, and several hydrocarbons. For 2alpha,3beta,23-trihydroxyolean-12-en-28-oic acid (2) and 3beta-O-acetylurs-12-en-28-oic acid (6) a first detailed assignment of ¹H NMR is presented.

Comparação do desempenho da lipase de candida rugosa imobilizada em suporte híbrido de polissiloxano-polivinilálcool empregando diferentes metodologias

The efficiency for immobilizing microbial Candida rugosa lipase on a hybrid matrix of polysiloxane polyvinyl alcohol, by adsorption, covalent coupling and encapsulation was compared. The activities of immobilized derivatives were evaluated using p-nitrophenylpalmitate (hydrolysis) and butyric acid and butanol (esterification) as substrates. Operational stability and storage tests were also performed. Among the procedures tested, the proposed matrix was efficient for immobilizing C. rugosa lipase by adsorption and covalent coupling techniques and unsuitable for encapsulation purposes. The results reveal that better catalytic properties in both aqueous and organic media were demonstrated by the covalent coupling POS-PVA immobilized lipase, including also satisfactory half-life and good storage stability.

Influência de métodos de digestão e massa de amostra na recuperação de nutrientes em resíduos orgânicos

The influence of nitric-perchloric, aqua regia, dry ashing and microwave digestion methods, in combination with 100 and 200 mg of sample, on the characterization and recovery of nutrients in samples of sludge sewage, poultry, swine, quail and bovine manures, organic compost, organic substrate and humic material were studied. Nitric-perchloric digestion with 200 mg samples recovered the higher nutrient contents. The nitric-perchloric method recovered also low levels of K. Dry ashing caused S volatilization and microwave digestion produced dark color extracts and this impaired S determination. Aqua regia recovered the lowest contents of nutrients in the organic residues evaluated.

Bioreduction of substituted a-tetralones promoted by Daucus carota root

The bioreduction of a series of substituted a-tetralones, carried out using Daucus carota root (carrot), afforded the corresponding homochiral a-tetralols in variable conversions (9 to 90%) and excellent enantiomeric excesses. Two of the assayed a-tetralones were resistant to the bioreduction conditions. The absolute configurations of four a-tetralols were assigned as being (S), by comparison to the (S)-enantiomers obtained by kinetic resolution promoted by CALB-catalysed acetylation. Additionally, the new 5-methoxy-6-methyl-1-tetralone was synthesized in seven steps from 3-methylsalicylic acid.

Otimização de método para determinação de flavonóis e flavonas em frutas por cromatografia líquida de alta eficiência utilizando delineamento estatístico e análise de superfície de resposta

This work optimized the HPLC conditions for the simultaneous determination of luteolin, apigenin, myricetin, quercetin and kaempferol in aglycone form, as well defined the best conditions for hydrolysis/extraction of these flavonoids in fruits, using the statistical central composite design and response surface analysis. A reverse phase method was developed using a gradient of methanol/water acidified with 0.3% formic acid as mobile phase and a photodiode array detector. The samples were extracted with methanol/water (50:50 v/v) at 90 ºC. The optimum time and HCl concentration varied for the different fruits investigated, demonstrating the necessity of optimizing these conditions for each fruit analyzed. Good recovery (87.1 to 96.3%), repeatability and linearity were obtained.

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.

Atividade biológica do lapachol e de alguns derivados sobre o desenvolvimento fúngico e em germinação de sementes

The natural quinones lapachol, α-lapachone and β-lapachone, and the synthetic derivative β-lapachone-3-sulfonic-acid were assayed for inhibition of fungal growth (Fusarium oxysporum) and germination of lettuce seeds (Lactuca sativa L.). β-Lapachone has the strongest activity as a germination inhibitor and lapachol shows no effect. β-Lapachone, followed by lapachol, are the most active in reducing fungal growth.

Steroidal and phenolic compounds from Sidastrum paniculatum (L.) Fryxell and evaluation of cytotoxic and anti-inflammatory activities

Sidastrum paniculatum (L.) Fryxell belongs to the family Malvaceae and is popularly known as "malva roxa" or "malvavisco". The phytochemical study of the hexane, CHCl3 and EtOAc phases from the crude ethanol extract of S. paniculatum led to the isolation of six compounds: a mixture of β-sitosterol and stigmasterol, 4-methoxy-3-hydroxybenzoic acid, 4-methoxy-3-hydroxybenzaldehyde, N-trans-feruloyltyramine and kaempferol-3-O-β-D-(6''-E-p -coumaroyl) glucoside. The structural identification of the compounds was made on the basis of spectroscopic methods such as IR, ¹H and 13C NMR with the aid of including two-dimensional techniques, besides comparison with literature data. The β-sitosterol and stigmasterol mixture showed a significant anti-inflammatory activity.

Determinação simultânea de ácido kójico e hidroquinona por espectrofotometria visível e calibração multivariada

A multivariate spectrophotometric method was developed for analysis of kojic acid/hydroquinone associations in skin whitening cosmetics. The method is based on the reaction between kojic acid and Fe3+ and on the reduction of Fe3+ by hydroquinone and further complexation of Fe2+ with 1,10-phenanthroline. The multivariate model was developed by Partial Least Squares Regression (PLSR), using 25 synthetic mixtures and mean-centered spectral data (350-380 nm). The use of 3 (kojic acid) and 2 (hydroquinone) latent variables permits the observation of mean errors of about 5% in the external validation phase.