Orthogonal Peptide-Templated Labeling Elucidates Lateral ETAR/ETBR Proximity and Reveals Altered Downstream Signaling

Fine-tuning of G protein-coupled receptor (GPCR) signaling is important to maintain cellular homeostasis. Recent studies demonstrated that lateral GPCR interactions in the cell membrane can impact signaling profiles. Here, we report on a one-step labeling method of multiple membrane-embedded GPCRs. Based on short peptide tags, complementary probes transfer the cargo (e. g. a fluorescent dye) by an acyl transfer reaction with high spatial and temporal resolution within 5 min. We applied this approach to four receptors of the cardiovascular system: the endothelin receptor A and B (ET_AR and ET_BR), angiotensin II receptor type 1, and apelin. Wild type-like G protein activation after N-terminal modification was demonstrated for all receptor species. Using FRET-competent dyes, a constitutive proximity between hetero-receptors was limited to ET_AR/ET_BR. Further, we demonstrate, that ET_AR expression regulates the signaling of co-expressed ET_BR. Our orthogonal peptide-templated labeling of different GPCRs provides novel insight into the regulation of GPCR signaling.     

Antioxidant properties,radical scavenging activity and biomolecule protection capacity of flavonoid naringenin and its glycoside naringin: a comparative study

BACKGROUND: This study was designed to evaluate and compare antioxidant capacity and radical scavenging activity of naringin and its aglycone by different in vitro assays. The effects of flavanones on lipid peroxidation, glutathione (GSH) oxidation and DNA cleavage were also assessed. RESULTS: The results showed that naringenin exhibited higher antioxidant capacity and hydroxyl and superoxide radical scavenger efficiency than naringin. Our results evidenced that glycosylation attenuated the efficiency in inhibiting the enzyme xanthine oxidase and the aglycone could act like a more active chelator of metallic ions than the glycoside. Additionally, naringenin showed a greater effectiveness in the protection against oxidative damage to lipids in a dose‐dependent manner. Both flavanones were equally effective in reducing DNA damage. However, they show no protective effect on oxidation of GSH. CONCLUSION: The data obtained support the importance of characterizing the ratio naringin/naringenin in foods when they are evaluated for their health benefits. Copyright © 2010 Society of Chemical Industry     

Analysis of bacterial community during the fermentation of pulque, a traditional Mexican alcoholic beverage, using a polyphasic approach

In this study, the characterization of the bacterial community present during the fermentation of pulque, a traditional Mexican alcoholic beverage from maguey (Agave), was determined for the first time by a polyphasic approach in which both culture and non-culture dependent methods were utilized. The work included the isolation of lactic acid bacteria (LAB), aerobic mesophiles, and 16S rDNA clone libraries from total DNA extracted from the maguey sap (aguamiel) used as substrate, after inoculation with a sample of previously produced pulque and followed by 6-h fermentation. Microbiological diversity results were correlated with fermentation process parameters such as sucrose, glucose, fructose and fermentation product concentrations. In addition, medium rheological behavior analysis and scanning electron microscopy in aguamiel and during pulque fermentation were also performed. Our results showed that both culture and non-culture dependent approaches allowed the detection of several new and previously reported species within the alpha-, gamma-Proteobacteria and Firmicutes. Bacteria diversity in aguamiel was composed by the heterofermentative Leuconostoc citreum, L. mesenteroides, L. kimchi, the gamma-Proteobacteria Erwinia rhapontici, Enterobacter spp. and Acinetobacter radioresistens. Inoculation with previously fermented pulque incorporated to the system microbiota, homofermentative lactobacilli related to Lactobacillus acidophilus, several alpha-Proteobacteria such as Zymomonas mobilis and Acetobacter malorum, other gamma-Proteobacteria and an important amount of yeasts, creating a starting metabolic diversity composed by homofermentative and heterofermentative LAB, acetic and ethanol producing microorganisms. At the end of the fermentation process, the bacterial diversity was mainly composed by the homofermentative Lactobacillus acidophilus, the heterofermentative L. mesenteroides, Lactococcus lactis subsp. lactis and the alpha-Proteobacteria A. malorum. After a 6-h fermentation, 83.27% of total sugars detected after inoculation were consumed (228.4 mM hexose equivalents) and a carbon (C) recovery of 66.18% in fermentation products was estimated. They were produced 284.4 mM C as ethanol, 71.5 mM C as acetic acid and 19 mM C as lactic acid, demonstrating the presence of homo- and heterofermentative, acetic and alcoholic metabolisms in the final product. It was also found, after hydrolysis, that the exopolysaccharide produced during the fermentation was mainly composed by fructose residues, probably inulin or levan.     

Fatty Acids and Metabolomic Composition of Follicular Fluid Collected from Environments Associated with Good and Poor Oocyte Competence in Goats

In goats, embryo oocyte competence is affected by follicle size regardless the age of the females. In previous studies we have found differences in blastocyst development between oocytes coming of small (<3 mm) and large follicles (>3 mm) in prepubertal (1–2 months-old) goats. Oocyte competence and Follicular Fluid (FF) composition changes throughout follicle growth. The aim of this study was to analyze Fatty Acids (FAs) composition and metabolomic profiles of FF recovered from small and large follicles of prepubertal goats and follicles of adult goats. FAs were analyzed by chromatography and metabolites by 1H-Nuclear Magnetic Resonance (1H-NMR) Spectrometry. The results showed important differences between adult and prepubertal follicles: (a) the presence of α,β-glucose in adult and no detection in prepubertal; (b) lactate, -N-(CH3)3 groups and inositol were higher in prepubertal (c) the percentage of Linolenic Acid, Total Saturated Fatty Acids and n-3 PUFAs were higher in adults; and (d) the percentage of Linoleic Acid, total MUFAs, PUFAs, n-6 PUFAs and n-6 PUFAs: n-3 PUFAs ratio were higher in prepubertal goats. Not significant differences were found in follicle size of prepubertal goats, despite the differences in oocyte competence for in vitro embryo production.     

Synthesis and structural characterization of an alternating double aquo-bridged and single cyano-bridged polymeric barium–iron complex: {[Ba2(phen)4(H2O)6Fe(CN)6]·Cl·2(phen)·3H2O}n

A new alternating double aquo-bridged and single cyano-bridged polymeric complex {[Ba₂(phen)₄(H₂O)₆Fe(CN)₆]·Cl·2(phen)·3H₂O}_n (1) (phen = 1,10-phenanthroline) has been synthesized and structurally characterized. In the crystal structure the two centrosymmetric [Ba₂(phen)₄(H₂O)₆] units are bridged through two trans CN groups of [Fe(CN)₆]³⁻ ion, which results in the formation of a zig-zag polymeric chain. In each [Ba₂(phen)₄(H₂O)₆] unit, the two Ba centers are joined by double aquo bridges. Both the Ba atoms are 9-coordinated with distorted mono-capped square antiprismatic geometry. An elaborate hydrogen bonding system holds the parallel polymeric chains together.     

Experimental evidence of an upper limit for hydrogen storage at 77 K on activated carbons

An upper limit for hydrogen storage at 77 K on activated carbons was clearly observed in the present experimental work. Such a limit is around 6.4 wt.%, i.e., close to the theoretical limit of 6.8 wt.%. Results of hydrogen storage were obtained in three independent laboratories using volumetric and gravimetric devices. Lab-made activated carbons (ACs) were found to have higher capacities than those of the commercial material AX-21. A maximum excess hydrogen storage capacity of 6.0 wt.% at 77 K and 4 MPa was obtained. This maximum was reduced to 0.6 wt.% at 298 K and 5 MPa. ACs with surface areas (S_BET) as high as 3220 m² g⁻¹ were prepared from chemical activation of anthracites with alkali (Na and K) hydroxides. At 77 K and 4 MPa, excess hydrogen storage capacity was directly correlated with S_BET for ACs having S_BET values lower than 2630 m²/g. Hydrogen uptake at 77 K also correlated with micropore volume and strongly depended on average pore diameter.     

Effect of glucose oxidase and mixing time on soluble and insoluble wheat flour protein fractions: Changes on SH groups and H2O2 consumption

Glucose oxidase (GOX) has been recognized as an alternative to substitute chemical oxidants in bread making, however the action exerted on wheat flour proteins by the H₂O₂ produced is still unclear. Looking for a better understanding of the role of GOX and H₂O₂ in the bread making process, the effect of varying GOX levels and mixing time on soluble and insoluble wheat flour protein fractions was studied. Results showed that GOX promoted a decrement in free SH groups in both proteins, being the most evident effect on the soluble fraction at the beginning of the mixing process, remaining almost constant afterwards. H₂O₂ was continuously produced during mixing, showing its largest consumption during the first 6 min without significant SH changes afterwards. Results suggest that H₂O₂ must be involved in other reactions as it remains in the sample; this also suggests that oxygen is not a limiting factor in this reaction.     

The crystal structures of three azonaphtharylamide pigments

The single crystal X-ray structures of three structurally related azonaphtharylamide pigments are reported. Two azo pigments derived from 4-amino-3-nitrotoluene as diazo component (1a and b) crystallize in the centrosymmetric space group P2₁/c while the third (1c), an analogue of 1b but derived from 2,5-dichloroaniline as diazo component, crystallizes in the chiral space group P2₁2₁2₁. The compounds adopt the ketohydrazone tautomeric forms with intramolecular, but no intermolecular, hydrogen bonding. The application performance of the products is discussed in relation to the molecular and crystal structures. X-ray powder diffraction, supported by FTIR spectroscopic and DSC analysis, demonstrate that pigment 1a shows polymorphism. The application performances of the two polymorphs of this pigment are compared.     

Interference of EDTA in the treatment of metal plating wastewater by biosorption

BACKGROUND: The applicability of biosorption for the treatment of metal plating wastewater is adversely affected by the presence of complexing agents. To investigate this limitation on the removal of copper(II) onto peat, batch and column experiments were carried out using EDTA as the model complexing agent. The influence of pH and copper(II):EDTA mass ratios were evaluated for copper(II) concentrations between 5 and 100 mg Cu(II) dm^?3. RESULTS: EDTA negatively affected the copper(II) uptake of peat for pH > 5. Batch and column experiments showed that copper(II)‐EDTA complexes were not sorbed by peat. The leaks of copper(II) detected from the beginning of the column operation matched the copper(II)‐EDTA concentration in the feed solutions. To overcome the interference of EDTA, a novel approach based on the combination of peat + activated carbon was proposed. Nearly complete removal of copper(II) was maintained over 70 h in the treatment of a solution containing 20 mg Cu(II) dm^?3 with 11% of copper(II)‐EDTA complexes. CONCLUSION: A new mass transport model coupling copper(II) speciation in the feed and mass transfer rate‐controlled process simulated the copper(II) breakthrough curves in the presence of EDTA and could be used to successfully predict the breakthrough point. This work demonstrated that biosorption can also be applied for the treatment of wastewater containing complexing agents with the proper combination of sorbent materials. © 2012 Society of Chemical Industry