Discovery of a New, Recurrent Enzyme in Bacterial Phosphonate Degradation: (R)‑1-Hydroxy-2-aminoethylphosphonate Ammonia-lyase

Phosphonates represent an important source of bioavailable phosphorus in certain environments. Accordingly, many microorganisms (particularly marine bacteria) possess catabolic pathways to degrade these molecules. One example is the widespread hydrolytic route for the breakdown of 2-aminoethylphosphonate (AEP, the most common biogenic phosphonate). In this pathway, the aminotransferase PhnW initially converts AEP into phosphonoacetaldehyde (PAA), which is then cleaved by the hydrolase PhnX to yield acetaldehyde and phosphate. This work focuses on a pyridoxal-5’-phosphate-dependent enzyme that is encoded in over 13 % of the bacterial gene clusters containing the phnW-phnX combination. This enzyme (which we termed PbfA) is annotated as a transaminase, but there is no obvious need for an additional transamination reaction in the established AEP degradation pathway. We report here that PbfA from the marine bacterium Vibrio splendidus catalyzes an elimination reaction on the naturally occurring compound (R)-1-hydroxy-2-aminoethylphosphonate (R-HAEP). The reaction releases ammonia and generates PAA, which can be then hydrolyzed by PhnX. In contrast, PbfA is not active towards the (S) enantiomer of HAEP or other HAEP-related compounds such as ethanolamine and D,L-isoserine, indicating a very high substrate specificity. We also show that R-HAEP (despite being structurally similar to AEP) is not processed efficiently by the PhnW-PhnX couple in the absence of PbfA. In sum, the reaction catalyzed by PbfA serves to funnel R-HAEP into the hydrolytic pathway for AEP degradation, expanding the scope and the usefulness of the pathway itself

Land use change in the high mountain belts of the central Apennines led to marked changes of the grassland mosaic

Aims High mountain pastures are hotspots of biodiversity, but grazing cessation and climate change are causing tall-grass encroachment and expansion of scrublands and forests. As part of biodiversity conservation efforts, grassland variation needs to be investigated at different spatial scales. We aimed to assess the landscape mosaic variation that occurred between 1988 and 2015 in the higher Mediterranean mountains. We investigated the recovery or land-degradation processes related to land use change, the effects of site condition, the impacts on grassland mosaic heterogeneity, and the threats to biodiversity. Location Sibillini Mountains (central Italy), over 1,650 m a.s.l. Methods We used two-step object-based supervised classification on Landsat 5 and 8 satellite images to analyze changes in landscape patterns and vegetation cover on formerly low-intensity pastures, by assessing the Normalized Difference Vegetation Index variation between 1988 and 2015. Twenty percent of the polygons obtained from segmentation were visually interpreted and assigned to five land cover classes. We generated a land use transition matrix and used Fourier Transforms to detect trends in variation of landscape mosaics and fragmentation. Results We observed prominent dynamics of the grassland mosaic leading to the homogenization of its structure through decreasing patch heterogeneity, especially on south-facing slopes. Grasslands shifted from open communities to dense pastures, with a reduction of scree and spread of tall grasses. The former trend could be understood as a recovery process reverting screes to conditions in equilibrium with local landform and climatic features, while the invasion of tall grasses is a land-degradation process that might lead to local species extinction and loss of habitat connectivity. Conclusions Pronounced changes in the large-scale landscape characteristics, mainly due to land use changes, of which scientists and managers of protected areas are not fully aware, are underway in the top mountain sectors of the study area

Diaminobenzidine photoconversion is a suitable tool for tracking the intracellular location of fluorescently labelled nanoparticles at transmission electron microscopy.

Chitosan-based nanoparticles (NPs) deserve particular attention as suitable drug carriers in the field of pharmaceutics, since they are able to protect the encapsulated drugs and/or improve their efficacy by making them able to cross biological barriers (such as the blood-brain barrier) and reach their intracellular target sites. Understanding the intracellular location of NPs is crucial for designing drug delivery strategies. In this study, fluorescently-labelled chitosan NPs were administered in vitro to a neuronal cell line, and diaminobenzidine (DAB) photoconversion was applied to correlate fluorescence and transmission electron microscopy to precisely describe the NPs intracellular fate. This technique allowed to demonstrate that chitosan NPs easily enter neuronal cells, predominantly by endocytosis; they were found both inside membrane-bounded vesicles and free in the cytosol, and were observed to accumulate around the cell nucleus

Finite mixture model-based classification of a complex vegetation system

To propose a Finite Mixture Model (FMM) as an additional approach for classifying large datasets of georeferenced vegetation plots from complex vegetation systems. Study area: The Italian peninsula including the two main islands (Sicily and Sardinia), but excluding the Alps and the Po plain. Methods: We used a database of 5,593 georeferenced plots and 1,586 vascular species of forest vegetation, created in TURBOVEG by storing published and unpublished phytosociological plots collected over the last 30 years. The plots were classified according to species composition and environmental variables using a FMM. Classification results were compared with those obtained by TWINSPAN algorithm. Groups were characterized in terms of ecological parameters, dominant and diagnostic species using the fidelity coefficient. Interpretation of resulting forest vegetation types was supported by a predictive map, produced using discriminant functions on environmental predictors, and by a non\u2010metric multidimensional scaling ordination. Results: FMM clustering obtained 24 groups that were compared with those from TWINSPAN, and similarities were found only at a higher classification level corresponding to the main orders of the Italian broadleaf forest vegetation: Fagetalia sylvaticae, Carpinetalia betuli, Quercetalia pubescenti-petraeae and Quercetalia ilicis. At lower syntaxonomic level, these 24 groups were referred to alliances and sub-alliances. Conclusions: Despite a greater computational complexity, FMM appears to be an effective alternative to the traditional classification methods through the incorporation of modelling in the classificatory process. This allows classification of both the co-occurrence of species and environmental factors so that groups are identified not only on their species composition, as in the case of TWINSPAN, but also on their specific environmental niche

Blood volume is improved by forced mild physical training in the motor and hippocampal cortex of old mice

The effect of mild forced physical training [1] (treadmill running 30 min a day, five days a week for 30 days at belt speed = 8 m/min, 0% incline) on the motor and hippocampal brain cortex was investigated in old (>24mo) mice by means of magnetic resonance imaging. The possible additive effect of physical training and testosterone [2] administration was also examined. Cortical thickness, quantitative transverse relaxation time (T2) maps, and regional cerebral blood volume (rCBV) were evaluated at baseline and after training. Results show that physical training alone induced significant increase of rCBV in both motor and hippocampal cortex. Cortex thickness and T2 maps were similar before and after training. Similar results were obtained in testosterone treated mice suggesting that testosterone does not add to physical training effect. This work provides first quantitative evidence that exercise initiated at old age is able to improve the hemodynamic status of the brain cortex in key regions for movement and cognition without inducing edema

A eutherian-specific microRNA controls the translation of Satb2 in a model of cortical differentiation

Cerebral cortical development is controlled by key transcription factors that specify the neuronal identities in the different layers. The mechanisms controlling their expression in distinct cells are only partially known. We investigated the expression and stability of Tbr1, Bcl11b, Fezf2, Satb2, and Cux1 mRNAs in single developing mouse cortical cells. We observe that Satb2 mRNA appears much earlier than its protein and in a set of cells broader than expected, suggesting an initial inhibition of its translation, subsequently released during development. Mechanistically, Satb2 30UTR modulates protein translation of GFP reporters during mouse corticogenesis. We select miR541, a eutherian-specific miRNA, and miR-92a/b as the best candidates responsible for SATB2 inhibition, being strongly expressed in early and reduced in late progenitor cells. Their inactivation triggers robust and premature SATB2 translation in both mouse and human cortical cells. Our findings indicate RNA interference as a major mechanism in timing cortical cell identities

An Eutherian-Specific microRNA Controls the Translation of Satb2 in a Model of Cortical Differentiation

Cerebral cortical development is controlled by key transcription factors that specify the neuronal identities in the different cortical layers. These transcription factors are crucial for the identity of the different neurons, but the mechanisms controlling their expression in distinct cells are only partially known. Here we investigate the expression and stability of the mRNAs of Tbr1, Bcl11b, Fezf2, Satb2 and Cux1 in single developing mouse cortical cells. We focus on Satb2 and find that its mRNA expression occurs much earlier than its protein synthesis and in a set of cells broader than expected, suggesting an initially tight control of its translation, which is subsequently de-repressed at late developmental stages. Mechanistically, Satb2 3\u2019UTR modulates protein translation of GFP reporters during mouse corticogenesis. By in vitro pull-down of Satb2 3\u2019UTR-associated miRNAs, we select putative miRNAs responsible for SATB2 inhibition, focusing on those strongly expressed in early progenitor cells and reduced in late cells. miR-541, an Eutherian-specific miRNA, and miR-92a/b are the best candidates and their inactivation triggers robust and premature SATB2 translation in both mouse and human cortical cells. Our findings indicate that RNA interference plays a major role in the timing of cortical cell identity and may be part of the toolkit involved in specifying supra-granular projection neurons