High concentrations of the melatonin metabolite, N-1-acetyl-N (2)-formyl-5-methoxykynuramine, in cerebrospinal fluid of patients with meningitis: a possible immunomodulatory mechanism

We evaluated the presence of the melatonin metabolite N-1-acetyl-N-2-formyl-5-methoxykynuramine (AFMK), in cerebrospinal fluid (CSF) of patients with viral meningitis (n = 20) and control samples (n = 8) and correlate AFMK levels with inflammatory markers such as cellularity, protein, tumor necrosis factor (TNF)-alpha, interleukin (IL)-8 and IL-1 beta levels. A portion of the CSF was extracted with dichloromethane (1:5) and analyzed by high-performance liquid chromatography (HPLC) under standardized conditions for AFMK. AFMK was detected in 16 of 20 CSF samples of patients with viral meningitis; the concentration of AFMK was found to be above the quantification limit (50 nmol/L) in six of these samples. AFMK was not detected in any of the eight control samples. The samples were classified into groups according to AFMK levels: undetectable (< 10 nmol/L, group I), detectable but below the quantification limit (< 50 nmol/L, group II), and quantified (> 50 nmol/L, group III). Group II presented the highest levels of proteins and IL-8, whereas group III showed the lowest levels of the inflammatory parameters. This study supports our hypothesis that inflammation favors the formation of AFMK and that this compound has immunomodulatory activity in vivo.

Optimized synthesis of the melatonin metabolite N(1)-acetyl-N(2)-formyl-5-methoxykynuramine (AFMK)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Metabolite fingerprint of "capim dourado" (Syngonanthus nitens), a basis of Brazilian handcrafts

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Determination of herbicides and a metabolite in human urine by liquid chromatography-electrospray ionization mass spectrometry

A method was developed to determine simazine, atrazine and their metabolite, 2-chloro-4,6-diamino-1,3,5-triazine, in urine. The presence of these herbicides in urine may reflect possible exposure to pesticides. Sample preparation involved protein precipitation and solid-phase extraction. The samples were analyzed by high-performance liquid chromatography-mass spectrometry. The detection limits were 0.4 mug/l and the analytes have a linear response in the interval 6-800 mug/l. The precision of the method was reflected in the RSD of <2.4% for the herbicides studied. Based on the detectable herbicide levels from spiked urine samples collected from unexposed volunteers, this method can be used to determine the low levels necessary for establishing reference values of the selected herbicides and the metabolite. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Dietary macronutrients and performance and plasma hormone and metabolite levels of broiler chickens fat by carbohydrate substitution

The present experiment was aimed to study the effects of an isocaloric substitution of fat by carbohydrate calories while maintaining the same CP content - on some endocrine parameters and key metabolites of the lipid (L), protein (CP) and carbohydrate (CHO) metabolism and muscle cell membrane integrity of broiler chickens. Ninety male broiler chicks (Cobb) were obtained at 1-day old and raised in an environmentally controlled room. Food and water was provided ad libitum. At day 7, the chicks were provided with the experimental diets: Control diet (CON: 20% CP; 9.1% L; 39.6% CHO), Low lipid diet (LowL: 22% CP; 3.36% L; 59.7% CHO), and low carbohydrate diet (LowCHO: 22% CP; 8.09% L; 46.7% CHO). T e chickens fed the CON manifested the best growth rate feed conversion compared to both other groups. The CO chickens showed the highest plasma T-3 Concentrations at 14 days of age as compared to both LowL and LowCHO chickens. At 35 days of age however, LowL chickens were characterized by the highest plasma T-3 levels. Overall, CON birds had the highest plasma T-4 levels (P < 0.0001) compared to LowL and LowCHO chickens. Plasma corticosterone levels were not affected by diet composition. Glucose levels increased towards the end of the experimental period, but there was no overall effect of diet composition on plasma glucose levels. Chickens fed the CON diet were consistently characterized by the highest plasma uric acid levels compared to LowL and LowCHO chickens. Irrespectively of diet, plasma triglyceride levels were high at 14 days of age, then decreased and finally increased again during the last week. This latter increase was most pronounced for the CON chickens and is congruous with their high fat deposition. At 28 days of age, a significant effect of diet was observed, as CON birds had lower plasma free fatty acid (FFA) levels compared to LowL but not to LowCHO chickens. However there was no overall effect of diet composition on plasma FFA levels. Creatine kinase (CK) activities increased significantly with age and were consistently higher in CON chickens. In conclusion, fast growth in broiler chickens is positively correlated with catabolism of protein from dietary or endogenous origin and (muscle) cell membrane disruption as reflected in high plasma uric acid levels and CK activity, respectively. The currently investigated substitution of fat by CHO had no marked effects on endocrine functioning and intermediary metabolism, but this does not exclude that larger substitutions between both nutrients can have an effect.

Comparative effects of lantadene A and its reduced metabolite on mitochondrial bioenergetics

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

High resolution ultra high pressure liquid chromatography-time-of-flight mass spectrometry dereplication strategy for the metabolite profiling of Brazilian Lippia species

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

Fiber fermentability effects on energy and macronutrient digestibility, fecal traits, postprandial metabolite responses, and colon histology of overweight cats

Considering the different potential benefits of divergent fiber ingredients, the effect of 3 fiber sources on energy and macronutrient digestibility, fermentation product formation, postprandial metabolite responses, and colon histology of overweight cats (Felis catus) fed kibble diets was compared. Twenty-four healthy adult cats were assigned in a complete randomized block design to 2 groups of 12 animals, and 3 animals from each group were fed 1 of 4 of the following kibble diets: control (CO; 11.5% dietary fiber), beet pulp (BP; 26% dietary fiber), wheat bran (WB; 24% dietary fiber), and sugarcane fiber (SF; 28% dietary fiber). Digestibility was measured by the total collection of feces. After 16 d of diet adaptation and an overnight period without food, blood glucose, cholesterol, and triglyceride postprandial responses were evaluated for 16 h after continued exposure to food. on d 20, colon biopsies of the cats were collected under general anesthesia. Fiber addition reduced food energy and nutrient digestibility. of all the fiber sources, SF had the least dietary fiber digestibility (P < 0.05), causing the largest reduction of dietary energy digestibility (P < 0.05). The greater fermentability of BP resulted in reduced fecal DM and pH, greater fecal production [g/(cat x d); as-is], and greater fecal concentration of acetate, propionate, and lactate (P < 0.05). For most fecal variables, WB was intermediate between BP and SF, and SF was similar to the control diet except for an increased fecal DM and firmer feces production for the SF diet (P < 0.05). Postprandial evaluations indicated reduced mean glucose concentration and area under the glucose curve in cats fed the SF diet (P < 0.05). Colon mucosa thickness, crypt area, lamina propria area, goblet cell area, crypt mean size, and crypt in bifurcation did not vary among the diets. According to the fiber solubility and fermentation rates, fiber sources can induce different physiological responses in cats, reduce energy digestibility, and favor glucose metabolism (SF), or improve gut health (BP).

Metabolite profiles of essential oils in citrus peels and their taxonomic implications

China is an important center of origin for the genus Citrus L. of the family Rutaceae and is rich in wild Citrus species. The taxonomy of Citrus has been a subject of controversy for more than a half century. We propose that the metabolite profiles of Chinese native Citrus species can be used for classification and understanding of the taxonomic relationships within the Citrus germplasm. In this study, triplicate gas chromatography-mass spectrometry (GC-MS) metabolite profiles of 20 Citrus species/varieties were acquired, including 10 native varieties originating in China. R-(+)-limonene, alpha-pinene, sabinene and alpha-terpinene were found to be major characteristic components of the essential oils analyzed in this study, and these compounds contributed greatly to the metabolic classification. The three basic species of the subgenus Eucitrus (Swingle's system), i.e., C. reticulata Blanco, C. medica L. and C. grandis Osb., were clearly differentiated based upon their metabolite profiles using hierarchical cluster analysis (HCA) and partial least square-discriminant analysis (PLS-DA). All the presumed hybrid genotypes, including sweet orange (C. sinensis Osb.), sour orange (C. aurantium L.), lemon (C. limon Burm.f.), rough lemon (C. jambhiri Lush.), rangpur lime (C. limonia Osb.) and grapefruit (C. paradisi Macf.), were grouped closely together with one of their suggested parent species in the HCA-dendrogram and the PLS-DA score plot. These results clearly demonstrated that the metabolite profiles of Citrus species could be utilized for the taxonomic classification of the genus and are complementary to the existing taxonomic evidence, especially for the identification and differentiation of hybrid species.

Curcumin pharmacokinetic and pharmacodynamic evidences in streptozotocin-diabetic rats support the antidiabetic activity to be via metabolite(s)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Automated analysis of lidocaine and its metabolite in plasma by in-tube solid-phase microextraction coupled with LC-UV for pharmacokinetic study

A sensitive, selective, and reproducible in-tube solid-phase microextraction and liquid chromatographic (in-tube SPME/LC-UV) method for determination of lidocaine and its metabolite monoethylglycinexylidide (MEGX) in human plasma has been developed, validated, and further applied to pharmacokinetic study in pregnant women with gestational diabetes mellitus (GDM) subjected to epidural anesthesia. Important factors in the optimization of in-tube SPME performance are discussed, including the draw/eject sample volume, draw/eject cycle number, draw/eject flow rate, sample pH, and influence of plasma proteins. The limits of quantification of the in-tube SPME/LC method were 50 ng/mL for both metabolite and lidocaine. The interday and intraday precision had coefficients of variation lower than 8%, and accuracy ranged from 95 to 117%. The response of the in-tube SPME/LC method for analytes was linear over a dynamic range from 50 to 5000 ng/mL, with correlation coefficients higher than 0.9976. The developed in-tube SPME/LC method was successfully used to analyze lidocaine and its metabolite in plasma samples from pregnant women with GDM subjected to epidural anesthesia for pharmacokinetic study.

Fiber fermentability effects on energy and macronutrient digestibility, fecal traits, postprandial metabolite responses, and colon histology of overweight cats

Considering the different potential benefits of divergent fiber ingredients, the effect of 3 fiber sources on energy and macronutrient digestibility, fermentation product formation, postprandial metabolite responses, and colon histology of overweight cats (Felis catus) fed kibble diets was compared. Twenty-four healthy adult cats were assigned in a complete randomized block design to 2 groups of 12 animals, and 3 animals from each group were fed 1 of 4 of the following kibble diets: control (CO; 11.5% dietary fiber), beet pulp (BP; 26% dietary fiber), wheat bran (WB; 24% dietary fiber), and sugarcane fiber (SF; 28% dietary fiber). Digestibility was measured by the total collection of feces. After 16 d of diet adaptation and an overnight period without food, blood glucose, cholesterol, and triglyceride postprandial responses were evaluated for 16 h after continued exposure to food. On d 20, colon biopsies of the cats were collected under general anesthesia. Fiber addition reduced food energy and nutrient digestibility. Of all the fiber sources, SF had the least dietary fiber digestibility (P < 0.05), causing the largest reduction of dietary energy digestibility (P < 0.05). The greater fermentability of BP resulted in reduced fecal DM and pH, greater fecal production [g/(cat x d); as-is], and greater fecal concentration of acetate, propionate, and lactate (P < 0.05). For most fecal variables, WB was intermediate between BP and SF, and SF was similar to the control diet except for an increased fecal DM and firmer feces production for the SF diet (P < 0.05). Postprandial evaluations indicated reduced mean glucose concentration and area under the glucose curve in cats fed the SF diet (P < 0.05). Colon mucosa thickness, crypt area, lamina propria area, goblet cell area, crypt mean size, and crypt in bifurcation did not vary among the diets. According to the fiber solubility and fermentation rates, fiber sources can induce different physiological responses in cats, reduce energy digestibility, and favor glucose metabolism (SF), or improve gut health (BP).

The fungal metabolite eugenitin as additive for Aspergillus niveus glucoamylase activation

Endophytic microorganisms live inside tissues of host plants apparently do not causing warning to them, and area promising source of bioactive molecules as antimicrobial and antitumoral drugs. In this work, we report the isolation of eugenitin from cultures of the endophyte Mycoleptodiscus indicus and its potential as additive for Aspergillus niveus glucoamylase activation. The glucoamylase hydrolytic activity increased twofold using 5 mM of eugenitin and this activation could be explained by the binding mode of eugenitin with the three-dimensional structure of glucoamylase. The in silica prediction of ligand binding sites revealed at least 9 possible interaction sites able to accommodate eugenitin on glucoamylase from Hypocrea jecorina. Besides, we evaluated the effect of pH and temperature on activity and stability, as well as in the hydrolysis of different substrates and kinetic parameters either in presence or absence of eugenitin. The results displayed by eugenitin as additive to glucoamylase activation are promising and provide novel perspectives for applications of fungal metabolites. (C) 2011 Elsevier B.V. All rights reserved.

Endo-xylanase GH11 activation by the fungal metabolite eugenitin

Eugenitin, a chromone derivative and a metabolite of the endophyte Mycoleptodiscus indicus, at 5 mM activated a recombinant GH11 endo-xylanase by 40 %. The in silico prediction of ligand-binding sites on the three-dimensional structure of the endo-xylanase revealed that eugenitin interacts mainly by a hydrogen bond with a serine residue and a stacking interaction of the heterocyclic aromatic ring system with a tryptophan residue. Eugenitin improved the GH11 endo-xylanase activity on different substrates, modified the optimal pH and temperature activities and slightly affected the kinetic parameters of the enzyme.