The tetraheme cytochrome from Shewanella oneidensis MR-1 shows thermodynamic bias for functional specificity of the hemes

Bacteria of the genus Shewanella contain an abundant small tetraheme cytochrome in their periplasm when growing anaerobically. Data collected for the protein isolated from S. oneidensis MR-1 and S. frigidimarina indicate differences in the order of oxidation of the hemes. A detailed thermodynamic characterization of the cytochrome from S. oneidensis MR-1 in the physiological pH range was performed, with data collected in the pH range 5.5–9.0 from NMR experiments using partially oxidized samples and from redox titrations followed by visible spectroscopy. These data allow the parsing of the redox and redox–protonation interactions that occur during the titration of hemes. The results show that electrostatic effects dominate the heme–heme interactions, in agreement with modest redox-linked structural modifications, and protonation has a considerable influence on the redox properties of the hemes in the physiological pH range. Theoretical calculations using the oxidized and reduced structures of this protein reveal that the bulk redox–Bohr effect arises from the aggregate fractional titration of several of the heme propionates. This detailed characterization of the thermodynamic properties of the cytochrome shows that only a few of the multiple microscopic redox states that the protein can access are significantly populated at physiological pH. On this basis a functional pathway for the redox activity of the small tetraheme cytochrome from S. oneidensis MR-1 is proposed, where reduction and protonation are thermodynamically coupled in the physiological range. The differences between the small tetraheme cytochromes from the two organisms are discussed in the context of their biological role.     

Bioavailability of Stabilised Ferrous Gluconate with Glycine in Fresh Cheese Matrix: a Novel Iron Compound for Food Fortification

Iron fortification of foods continues to be one of the preferred ways of improving the iron status of the population. Dairy product is a common product in the diet; therefore, it is a plausible vehicle for iron fortification. This study aims to investigate the bioavailability of ferrous gluconate stabilised with glycine (FGSG) in a fresh cheese fortified with zinc. The iron bioavailability of fresh cheese fortified with either FGSG and with or without zinc and FGSG in aqueous solution and a water solution of ferrous ascorbate (reference dose) was studied using double radio iron (⁵⁵Fe and ⁵⁹Fe) erythrocyte incorporation in 15 male subjects. All subjects presented with normal values for iron status parameters. The geometric mean of iron bioavailability for the water solution of FGSG was 38.2 %, adjusted to 40 % from reference doses (N.S.). Iron bioavailability in fresh cheese fortified with Ca and Zn was 15.4 % and was 23.1 % without Zn, adjusted to 40 % from reference doses (N.S.). The results of the present study show that the novel iron compound ferrous gluconate stabilised with glycine in a fresh cheese matrix is a good source of iron and can be used in iron fortification programmes.     

Modification of the ferrozine technique to analyze iron contents in different foods

A methodology for the determination of iron in foods fortified with this element or in nutritional products is important and has to be sensitive and rapid. In developing countries, an inexpensive and reliable methodology is also required. For this purpose, the Gordon’s Ferrozine technique was slightly modified and assayed with yogurt, dry powdered milk, and cereal mixtures, all of them fortified with iron, using an internal standard as the reference methodology. The obtained results demonstrate a close correlation between the standard curve interpolation method and the internal standard reference method (correlation coefficient r² = 0.9950) in a wide range of concentrations. The slope (0.9998 ± 0.0040) demonstrates that both procedures measure equal amounts of iron. The conclusion is that the proposed technique is a reliable, practical, and inexpensive methodology for iron determination in different foods fortified with iron.     

Genetic differentiation among three neighboring Brazil-cherry (Eugenia uniflora L.) populations within the Brazilian Atlantic rain forest

Eugenia uniflora L. (pitanga) is widely distributed in tropical areas. It is present in coastal vegetation from Ceara, in northeastern Brazil, to Rio Grande do Sul, at the southern tip of the country. Eugenia uniflora is of ecological importance, both as colonizing species on disturbed land and as food supplier for a wide variety of insects, birds and mammals. Pitanga plays a role in the maintenance of shrubby coastal ecosystems, especially at disturbed sites, and in 'restinga' ecosystems, at the interface between low forest and strand vegetation. To investigate the genetic diversity residing within the species, three neighboring populations at a distance of less than 24 km from each other, with varying degrees of human impact, were studied. The level of genetic diversity within and between populations was assessed with amplified fragment length polymorphism (AFLP) methodology. A total of 532 AFLP markers were analyzed in 66 individual trees. The polymorphism level varied from 61.2 to 96% depending on the primer combination used. Intra- and inter-population genetic diversity analysis showed that more than 88% of the variation resided within the populations, with a G_st of 0.123. Nevertheless, using neighbor joining (NJ) and principal component analysis (PCA), on the genetic distance (GD) data, permitted the three analyzed populations to be differentiated.     

Infraspecific chemical variability of the leaf essential oil of Juniperus phoenicea var. turbinata from Portugal

The essential oil composition of 68 individual plants of Juniperus phoenicea from Portugal was investigated by GC, GC-MS and ¹³C NMR. -Pinene, β-phellandrene, -terpinyl acetate and myrcene were found to be the main constituents. Botanical and chemical data as well as phytogeographical distribution indicate J. phoenicea var. turbinata as the unique subspecies occurring in Portugal. Nevertheless, this taxon exhibits chemical polymorphism. The results of the oil compositions were processed by hierarchical clustering and principal component analysis allowing to establish three groups of essential oils differentiated by the content of -pinene, β-phellandrene and -terpinyl acetate.     

Acidianus ambivalens type-II NADH dehydrogenase: genetic characterisation and identification of the flavin moiety as FMN

The thermoacidophilic archaeon Acidianus ambivalens contains a monomeric 47 kDa type-II NADH dehydrogenase (NDH), which contains a covalently bound flavin. In this work, by a combination of several methods, namely (31)P-nuclear magnetic resonance and fluorescence spectroscopies, it is proven that this enzyme contains covalent FMN, a novelty among this family of enzymes, which were so far thought to mainly have the flavin dinucleotide form. Discrimination between several possible covalent flavin linkages was achieved by spectral and fluorescence experiments, which identified an 8alpha-N(1)-histidylflavin-type of linkage. Analysis of the gene-deduced amino acid sequence of type-II NDH showed no transmembranar helices and allowed the definition of putative dinucleotide and quinone binding motifs. Further, it is suggested that membrane anchoring can be achieved via amphipatic helices.     

Determination of the orientation of the axial ligands and of the magnetic properties of the haems in the tetrahaem ferricytochrome from Shewanella frigidimarina

The unambiguous assignment of the nuclear magnetic resonance (NMR) signals of the alpha-substituents of the haems in the tetrahaem cytochrome isolated from Shewanella frigidimarina NCIMB400, was made using a combination of homonuclear and heteronuclear experiments. The paramagnetic (13)C shifts of the nuclei directly bound to the porphyrin of each haem group were analysed in the framework of a model for the haem electronic structure. The analysis yields g-tensors for each haem, which allowed the assignment of some electron paramagnetic resonance (EPR) signals to specific haems, and the orientation of the magnetic axes relative to each haem to be established. The orientation of the axial ligands of the haems was determined semi-empirically from the NMR data, and the structural results were compared with those of the homologous tetrahaem cytochrome from Shewanella oneidensis MR-1 showing significant similarities between the two proteins.     

Microalgal biomass quantification from the non-invasive technique of image processing through red–green–blue (RGB) analysis

Journal of Applied Phycology - Continuous monitoring of biomass concentration in microalgae cultures is essential and one of the most important parameters to measure in this field. This study aims...     

Luminal iron levels govern intestinal tumorigenesis after Apc loss in vivo

It is clear from epidemiological studies that excess iron is associated with increased risk of colorectal cancer; however, questions regarding the mechanism of how iron increases cancer risk, the source of the excess iron (circulating or luminal), and whether iron reduction represents a potential therapeutic option remain unanswered. In this study, we show that after Apc deletion, the cellular iron acquisition proteins TfR1 and DMT1 are rapidly induced. Conversely, restoration of APC reduces cellular iron due to repression of these proteins. To test the functional importance of these findings, we performed in vivo investigations of the impact of iron levels on intestinal tumorigenesis. Strikingly, depletion of luminal (but not systemic) iron strongly suppressed murine intestinal tumorigenesis, whereas increased luminal iron strongly promoted tumorigenesis. Taken together, our data definitively delineate iron as a potent modifier of intestinal tumorigenesis and have important implications for dietary iron supplementation in patients at high risk of colorectal cancer.