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An FHA phosphoprotein recognition domain mediates protein EmbR phosphorylation by PknH, a Ser/Thr protein kinase from Mycobacterium tuberculosis.

In bacteria, regulatory phosphorylation of proteins at serine and/or threonine residues by Ser/Thr protein kinase (STPK) is an emerging theme in prokaryotic signaling, particularly since the prediction of the occurrence of several STPKs from genome sequencing and sequence surveys. Here we show that protein PknH possesses an autokinase activity and belongs to the large STPK family found in Mycobacterium tuberculosis. Evidence is presented that PknH can also phosphorylate EmbR, a protein suspected to modulate the level of arabinosyltransferase activity involved in arabinan biosynthesis of arabinogalactan, a key molecule of the mycobacterial cell wall. Interestingly, EmbR possesses an FHA (forkhead-associated) domain, a newly described phosphoprotein recognition domain, which plays an essential role in PknH-EmbR interaction and phosphorylation of EmbR by PknH. It is demonstrated that mutation of each of three particular residues of this FHA domain, Arg312, Ser326, and Asn348, totally abolishes the PknH-mediated phosphorylation of EmbR, thus highlighting the critical role of this domain in the direct interaction between EmbR and PknH.

Bacterial Proteins↗

Retention of trace metals by solidified/stabilized wastes: assessment of long-term metal release.

Toxic elements found in wastes may have a negative impact on the environment, especially through the contamination of groundwater and plants. To reduce their mobility and availability, French regulations mandate the solidification and stabilization of toxic wastes. Many methods to stabilize and solidify wastes exist, among them the Ecofix process which employs low cost materials and consists of mixing wastes with lime, aluminum hydroxide, and silica. To evaluate the long-term behavior of solidified/stabilized (S/S) samples, their alteration under saturated conditions was studied in a water extractor, a Soxhlet-like device, used to follow the weathering of rocks. Kinetic measurements have shown that the release of Fe, Pb, Cd, Cr, and Cu was very slow, indicating a strong retention of these elements by the S/S materials prepared by the Ecofix process. To elucidate the mechanisms of retention of the trace metals, the mineral phases that existed in the S/S samples throughout and at the end of the extraction runs were studied by X-ray diffraction and by infrared and nuclear magnetic resonance spectroscopies. Scanning electron microscopic (SEM) examinations and electron microprobe analyses of the S/S samples were also performed at different stages of weathering. These observations revealed that assorted calcium silicate hydrates (C-S-H) were the predominant phases in the S/S preparations and that gradual alterations occurred in the structure of the investigated materials. The overall Ca/Si ratio of the C-S-H phases decreased as the enhanced alteration progressed. Although trace metals in oxide, hydroxide, and carbonate forms were found in the S/S materials, the bulk of the trace metals was incorporated in the matrix of the C-S-H phases.

Aluminum Hydroxide↗

High resolution electron microscopy structural studies of organo-clay nanocomposites.

Engineering of clay nanocomposite materials by modification of their surfaces can enable the control of retention, transport, and persistence of toxic chemicals in the geosystem. The properties and interactions of clay nanocomposites have been widely studied, but little information exists on their microstructure at a range of scale extending down to atomic dimensions. The pairing of Na-montmorillonite clay with organic cations as well as with the herbicide fluridone, chosen as a model for an organic pollutant, was studied. Three organic cations were selected: hexadecyltrimethylammonium, benzyltrimethylammonium, and benzyltriethylammonium at 0%, 60%, and 100% of cation exchange capacity (CEC) loadings. A detailed microstructural analysis of the organo-clay nanocomposites and of the fluridone nanocomposites was undertaken by high-resolution transmission electron microscopy (HRTEM) and X-ray energy-dispersive spectroscopy (EDS). Morphological observations and chemical analyses were performed simultaneously on the same sample. The combined HRTEM and EDS measurements strongly suggest (a) heterogeneous local intercalation of the organic cations manifested by a range in the measured d001 spacing, implying random expansion of the clay layered structure with increased loading of the organic cations; (b) intercalation within the external layers, which is thoroughly influenced by local defect microstructure and/or edge availability of the montmorillonite nanoparticles as well as by the molecular structure of the intercalating organic cation. Additional intercalation of fluridone molecules did not affect the structure (d001 spacing) of the organo-clay nanocomposites.

Aluminum Silicates↗

Evaluation of the representativeness of the odor of cooked mussel extracts and the relationship between sensory descriptors and potent odorants.

The representativeness of the odor of mussel extract was assessed after each step of the distillation-extraction-concentration process. Results showed that the whole process was convenient for cooked mussels, but the extract was representative only when it was reincorporated into a suitable matrix such as water. Sensory and gas chromatography-olfactometry (GC-O) analyses were then performed on representative extracts of wild and bouchot mussels. Most of the sensory attributes were related to odors detected during olfactometry. Methional and (Z)-4-heptenal were two of the most potent odorants of mussels and, thus, were identified as the major contributors to the characteristic boiled potato-like odor of cooked mussels distinguished during sensory analysis. The sulfury note, highlighted for wild mussels during sensory analysis, could be linked to dimethyl disulfide, which was significantly more perceived in wild mussels by GC-O. Dimethyl disulfide could then be considered to be a characteristic compound of wild mussels.

Animals↗

Identification of the main odor-active compounds in musts from French and Romanian hybrids by three olfactometric methods.

Three olfactometric methods (frequency of detection, time--intensity method, and aroma extract dilution analysis) were used to evaluate the main odorants of three musts obtained from French--Romanian hybrids (Valerien, Admira, and Brumariu). The three methods allow detection of the same odor-active compounds. The results obtained from these methods were closely related. Nineteen odor-active compounds were detected, and 13 were identified. The three methods showed the importance of an unidentified compound with a grape and grape juice aroma note in the three musts. Among the other compounds, 3-hexen-1-al, (E,Z)-2,6-nonadien-1-ol, and 1-ccten-3-one seemed to contribute actively to the odor of Valerien must. 3-(Methylthio)propanal and hexanal were contributors to the Admira and Brumariu odor. Phenylacetaldehyde was one of the main odor-active compounds in must from Admira.

Chromatography, Gas↗

Aroma extracts from oyster Crassostrea gigas: comparison of two extraction methods.

The study of the aroma of oysters is of great economic interest in France because it enables their organoleptic quality to be verified. The aim of this study is to optimize the extraction methods of the volatile compounds of oysters Crassostrea gigas in order to obtain an extract with an odor as close as possible to that of the original oysters'. Oyster aroma is rarely studied, and its sensory profile has not been investigated to date. Two extraction methods were studied: vacuum hydrodistillation carried out at 20 degrees C with noncrushed oyster using ultrapure water and dynamic headspace carried out using noncrushed oyster during a 30 min purge. They were compared with regard to their sensory characteristics by a panel of seven judges, all trained in seafood aroma recognition. This study has shown that vacuum hydrodistillation is the better method to obtain an extract closest in aroma to the oyster reference.

Animals↗

Comparison of two microalgal diets. 1. Influence on the biochemical and fatty acid compositions of raw oysters (Crassostrea gigas).

Oyster farming in France is a traditional activity. Each year, 149 000 tons of oysters are fattened before being sold. More and more oyster farmers supplement the diet of oysters by microalgae to optimize the fattening process and to improve both the growth and flesh quality of oysters. In the present study, oysters were supplemented by two microalgae: Skeletonema costatum and Tahitian isochrysis clone. The ash, protein, carbohydrate (including glycogen), and lipid contents were analyzed. The growth of oysters was improved when they were fattened with microalgae, leading to an increase in the condition index. The chemical composition of oysters was influenced by the chemical composition of the microalgae, especially the level of glycogen, which was significantly increased for oysters supplemented by microalgae. The fatty acid profile of oysters fattened by microalgae was positively correlated with the fatty acid profiles of S. costatum and T. isochrysis. These results show the effectiveness of supplementation by microalgae on the growth and on the biochemical composition (glycogen and fatty acids especially) of oysters.

Animals↗

Optimization of dynamic headspace extraction of the edible red algae Palmaria palmata and identification of the volatile components.

A new extraction method was applied to the volatile compounds of Palmaria palmata. Dynamic headspace was optimized according to an experimental design, and descriptive sensory analysis and intensity and similarity tests were performed for each extract to assess their respective representativeness. Results showed that extract obtained with crushed algae after a 30 min purge was the most representative. GC-MS analysis was then performed on this extract to identify the volatile components. Seven halogenated compounds, seven aldehydes, two ketones, three alcohols, and four miscellaneous compounds were identified. Among them, halogenated compounds were the most characteristic of red algae, and more particularly, iodoethane and iodopentane, which had yet been found in other seaweeds.

Aldehydes↗

Diffusion coefficient of CO(2) molecules as determined by (13)C NMR in various carbonated beverages.

In this paper, the NMR technique was used, for the first time, to accurately determine the diffusion coefficient D of CO(2)-dissolved molecules in various carbonated beverages, including champagne and sparkling wines. This parameter plays an important role concerning the bubble growth during its rise through the liquid (see ref 3). The diffusion coefficient of CO(2)-dissolved molecules D was compared with that deduced from the well-known Stokes-Einstein equation and found to significantly deviate from the general trend expected from Stokes-Einstein theory, i.e, D(SE) proportional, variant 1/eta, where D(SE) is the Stokes-Einstein diffusion coefficient and eta the viscosity of the liquid medium.

Carbon Dioxide↗

Bound ferulic acid from bran is more bioavailable than the free compound in rat.

Ferulic acid (FA) is reported as a good antioxidant absorbed by human or rat but only few data deal with the influence of the food matrix on its bioavailability and with its potential protection against cardiovascular diseases and cancer. Wheat bran is used as a source of ferulic acid, the compound being mainly bound to arabinoxylans of the plant cell walls. Pharmacokinetic profiles of FA and its metabolites are established in rats. Free and conjugated FA quickly appear in plasma, reach a plateau 1 h after intake and remain approximately constant at 1 microM up to 24 h. 2.3% of FA are eliminated in urine. Compared with results obtained after intake of free FA, the presence of FA-arabinoxylans bonds in the food matrix increases the occurrence time of FA in the organism and decreases the level of urinary excretion in 24 h. Nevertheless, sulfated FA is still the main plasmatic form. The antioxidant activity of plasmas of rats fed with a standard diet (containing no FA), pure ferulic acid (5.15 mg FA/kg bw) or bran (4.04 mg FA/kg bw) are measured in an ex vivo test using AAPH as free radical inducer. Plasmas of rats fed with bran show a better antioxidant activity than the control group and the pure FA supplemented group, increasing the resistance of erythrocytes to hemolysis by factors of 2 and 1.5, respectively. These results show the good bioavailability of FA from bran and its potential efficiency to protect organism against pathology involving radical steps of development.

Animals↗

Optimization of extraction of apple aroma by dynamic headspace and influence of saliva on extraction of volatiles.

The dynamic headspace procedure of aroma extraction was optimized on Gala apples (Malus domestica). Two parameters affecting the extractability of compounds were studied: temperature and purge time. The influence of artificial saliva was also included. An increase in purge time and temperature caused an increase in the extraction of volatiles from the apple matrix. The optimum point of extraction was 40 degrees C and 70 min of purge. The study also showed that the addition of saliva influenced the extraction of volatile compounds, but this effect was different from one compound to another. To verify that the headspace extracts presented a global odor representativeness of fresh apple under these conditions of extraction, eight assessors compared the odor of extracts with fresh fruit odor for three different cultivars. With regard to the sensory profiles of extracts, the optimal conditions of extraction were suitable for extraction of volatile compounds, even if cooked apple odor appeared in some extracts. The similarity marks of extracts were low but acceptable.

Fruit↗

Influence of rearing conditions on the volatile compounds of cooked fillets of Silurus glanis (European catfish).

Volatile compounds of cooked fillets of Silurus glanis reared under two conditions occurring in France were studied. They were extracted by dynamic headspace, identified by gas chromatography/mass spectrometry, and quantified by gas chromatography-flame ionization detection. Odor active volatile compounds were characterized by gas chromatography-olfactometry. Sixty volatile compounds were detected in dynamic headspace extracts, among which 33 were odor active. Rearing conditions affected their estimated concentrations and their odor intensities, but very few qualitative differences were exhibited (only seven volatile compounds were concerned). A good correlation between quantitative and olfactometric results is shown. 2-Methylisoborneol and (E)-2-hexenal were less represented in OUTDOOR extracts, while 2-butanone was less represented in INDOOR extracts. In addition, olfactometric results can be closely related to those previously obtained by sensory analysis. Boiled potato sensory odor of the silurus cooked fillets can be related to (Z)-4-heptenal and methional, and buttery odor can be related to 2,3-butanedione, an unknown compound (RI = 1010), and 2,3-pentadione.

Animals↗

Characterization of odor-active volatiles in apples: influence of cultivars and maturity stage.

The aroma and texture of three different apple cultivars, harvested at three maturity stages, were analyzed by sensory and instrumental analysis. The emphasis was on the identification of the most potent odorant volatiles, and the challenge was to separate the few most important flavor compounds, which may be trace chemicals, from the vast number of nonodorant compounds present in apple aroma extracts. Thirty-six odorant compounds were detected, 24 of which were common to all extracts. A significant correlation coefficient was found between the aroma intensity scores and overall quantity of the odorant volatiles, which shows that the development of sensory aroma is similar to that of odorant volatiles. This study also showed that the parameters measured by penetrometry and compression were highly correlated with sensory textural attributes. The determination of the optimal maturity stage for different apple cultivars by the usual parameters, such as color, diameter, total soluble solids, and titrable acidity, may not be sufficient to determine the optimal sensory quality for consumers. Moreover, the sensory quality of fruits changes during maturation in a different way from one cultivar to another, and this should be taken into account.

Fruit↗

Comparison of odor-active volatile compounds of fresh and smoked salmon.

The odorant volatile compounds of raw salmon and smoked salmon have been investigated by two gas chromatography-olfactometry methods (frequency detection and odorant intensity) and gas chromatography-mass spectrometry. After simultaneous steam distillation-solvent extraction with diethyl ether and the recovery of the aromatic extract in ethanol, qualitative olfactometric characterization and identification followed by a quantitative assessment of the odorant volatile compounds were carried out. The origin of many odorant compounds of smoked salmon can be attributed to wood smoke. Another part of smoked salmon aroma is due either to the odorant compounds of the raw fish flesh or to an evolution of fish flesh aroma thanks to the smoking process conditions. Forty-nine odorant compounds have been identified in fresh salmon and 74 in smoked salmon. Carbonyl compounds, such as heptanal or (E,Z)-2,6-nonadienal, show a high detection frequency and odorant intensity in unsmoked fish, giving the flesh its typical fishy odor. For smoked salmon, phenolic compounds, such as cresol or guaiacol, and furanic compounds seem to be responsible for the smoked odor.

Animals↗

Characterization of aroma-active compounds in rainbow trout (Oncorhynchus mykiss) eliciting an off-odor.

The aroma-active and off-flavor compounds of cooked rainbow trout (Oncorhynchus mykiss) were analyzed by sensory and instrumental analyses. Sensory analysis shows that the aromatic extract obtained by vacuum steam distillation was representative of rainbow trout odor. To obtain more information on odorants of volatile compounds, analyses were conducted on two gas chromatography columns of different polarities (DB-5 and DB-Wax). The results of the gas chromatography-olfactometry analysis showed that 38 odorous compounds were perceived when the DB-5 column was used and 36 with the DB-Wax column. Of these, 31 with the DB-5 and 28 with the DB-Wax were identified. (E)-2-Nonenal, 2-ethyl-1-hexanol, 2-methylisoborneol, geosmin, 2-methylnaphthalene, and 8-heptadecene were described as off-flavor compounds by the sniffing assessors. The most powerful off-flavor compounds identified in the extract were 2-methylisoborneol and geosmin, which were described as strong musty and earthy odors, respectively.

Adult↗

Critical comparison of three olfactometric methods for the identification of the most potent odorants in cooked mussels (Mytilus edulis).

Three olfactometric methods (olfactometry global analysis, OSME, and AEDA) were compared to evaluate the main impact odorants of cooked mussels. The results obtained from these methods were very similar and well correlated. On the basis of the three techniques, 42 odor-active compounds were detected and 28 were identified. Among these compounds, 6 odorants seem to contribute actively to the aroma of mussels: 2,3-butanedione (4) (buttery, caramel-like odor), (Z)-4-heptenal (14) (boiled potato-like odor), (E)-2-penten-1-ol (17) (mushroom-like odor), 2-ethylpyrazine (19) (nutty odor), methional (25) (boiled potato-like odor), and (E,E)-2,4-octadienal (32) (cucumber-like odor).

Aldehydes↗

Aroma of fresh oysters Crassostrea gigas: composition and aroma notes.

In contrast to many foods, very little is known about the aroma of fresh oysters. This study deals with the relationship between extracted volatiles of oysters and their olfactory properties. Nearly 50 volatiles were identified: most of them were principally related to fatty acid oxidation (86%) and particularly to n-3 polyunstaturated fatty acid oxidation (66%). Only one volatile arose from amino acid degradation. Panelists detected 42 odors by sniffing. Among them, only 12 odors were definitely attributed to identified volatile. These odors were green/sulfur/crustacean, mushroom/citrus, and marine/cucumber notes and were attributed to dimethyl sulfide, 1-penten-3-one, hexanal, (2,4)-E,E-heptadienal, 1-octen-3-one, 1-octen-3-ol, 6-methyl-5-hepten-3-one, octanal, (E,Z)-2,6-nonadienal, (E)-2-octenal, and decanal, respectively.

Animals↗

New N alpha-guanidinobenzoyl derivatives of hirudin-54-65 containing stabilized carboxyl or phosphoryl groups on the side chain of phenylalanine-63.

We report on the synthesis and pharmacological properties of a new series of thrombin inhibitors derived from hirudin carboxyl-terminal fragments. Two (arylphosphono)phenylalanines, p-PO3H2-L-Phe1 and m-PO3H2-L-Tyr, and one (carboxymethyl)phenylalanine, p-CH2COOH-L-Phe, were prepared and incorporated into position 63 of the modified hirudin's C-terminal dodecapeptide using the Fmoc solid-phase synthesis strategy. Substitution by any one of the residues led to very active analogs which doubled the thrombin time at low micromolar concentration (Ctt2) in vitro (1 microM < Ctt2 < 3 microM) and potently increased the activated partial thromboplastin time (APTT) ex vivo. These compounds displayed a higher potency in vitro and a longer duration of action in vivo than both the corresponding sulfated or phosphorylated tyrosine counterparts.

Amino Acid Sequence↗