Design, synthesis, and biological evaluation of new (2E,6E)-10-(dimethylamino)-3,7-dimethyl-2,6-decadien-1-ol ethers as inhibitors of human and Trypanosoma cruzi oxidosqualene cyclase

New dimethylamino truncated squalene ether derivatives containing a different aromatic moiety (phenyl, naphthyl, and biphenyl) or a simple alkyl (n-hexylic) group were synthesized as inhibitors of the oxidosqualene cyclase (OSC) and of the sterol biosynthetic pathway. The activity against human OSC was compared with the activity against the OSCs of pathogenic organisms such as Pneumocystis carinii and Trypanosoma cruzi. The phenyl derivative was the most potent inhibitor of T. cruzi OSC.     

Endogenous microRNA can be broadly exploited to regulate transgene expression according to tissue, lineage and differentiation state

We have shown previously that transgene expression can be suppressed in hematopoietic cells using vectors that are responsive to microRNA (miRNA) regulation. Here we investigate the potential of this approach for more sophisticated control of transgene expression. Analysis of the relationship between miRNA expression levels and target mRNA suppression suggested that suppression depends on a threshold miRNA concentration. Using this information, we generated vectors that rapidly adjust transgene expression in response to changes in miRNA expression. These vectors sharply segregated transgene expression between closely related states of therapeutically relevant cells, including dendritic cells, hematopoietic and embryonic stem cells, and their progeny, allowing positive/negative selection according to the cells' differentiation state. Moreover, two miRNA target sites were combined to restrict transgene expression to a specific cell type in the liver. Notably, the vectors did not detectably perturb endogenous miRNA expression or regulation of natural targets. The properties of miRNA-regulated vectors should allow for safer and more effective therapeutic applications.     

Constitutive presence of antibiotic resistance genes within the bacterial community of a large subalpine lake

The fate of antibiotic resistance genes (ARGs) in environmental microbial communities is of primary concern as prodromal of a potential transfer to pathogenic bacteria. Although of diverse origin, the persistence of ARGs in aquatic environments is highly influenced by anthropic activities, allowing potential control actions in well‐studied environments. However, knowledge of abundance and space–time distribution of ARGs in ecosystems is still scarce. Using quantitative real‐time PCR, we investigated the presence and the abundance of twelve ARGs (against tetracyclines, β‐lactams, aminoglycosides, quinolones and sulphonamides) at different sampling sites, depths and seasons, in Lake Maggiore, a large subalpine lake, and in the area of its watershed. We then evaluated the correlation between each ARG and a number of ecological parameters in the water column in the deepest part of the lake. Our results suggest the constitutive presence of at least four ARGs within the bacterial community with a high proportion of bacteria potentially resistant to tetracyclines and sulphonamides. The presence of these ARGs was independent of the total bacterial density and temperature. The dynamics of tet(A) and sulII genes were, however, positively correlated with dissolved oxygen and negatively to chlorophyll a, suggesting that the resistant microbes inhabit specific niches. These observations indicate that the lake is a reservoir of antibiotic resistances, highlighting the need of a deeper understanding of the sources of ARGs and the factors allowing their persistence in waters.     

A novel autoregulatory loop between the Gcn2-Atf4 pathway and L-Proline metabolism controls stem cell identity

Increasing evidence indicates that metabolism is implicated in the control of stem cell identity. Here, we demonstrate that embryonic stem cell (ESC) behaviour relies on a feedback loop that involves the non-essential amino acid L-Proline (L-Pro) in the modulation of the Gcn2-Eif2α-Atf4 amino acid starvation response (AAR) pathway that in turn regulates L-Pro biosynthesis. This regulatory loop generates a highly specific intrinsic shortage of L-Pro that restricts proliferation of tightly packed domed-like ESC colonies and safeguards ESC identity. Indeed, alleviation of this nutrient stress condition by exogenously provided L-Pro induces proliferation and modifies the ESC phenotypic and molecular identity towards that of mesenchymal-like, invasive pluripotent stem cells. Either pharmacological inhibition of the prolyl-tRNA synthetase by halofuginone or forced expression of Atf4 antagonises the effects of exogenous L-Pro. Our data provide unprecedented evidence that L-Pro metabolism and the nutrient stress response are functionally integrated to maintain ESC identity.Naturally occurring amino acids are emerging as key players in the regulation of the phenotypic plasticity of stem cells.^{1, 2, 3, 4, 5} Indeed, exogenously provided threonine and methionine, two essential amino acids (EAAs), regulate self-renewal and differentiation of pluripotent stem cells.² Moreover, exogenously provided L-Proline (L-Pro), a non-essential amino acid (NEAA), induces mouse ESCs towards an embryonic stem cell-to-mesenchymal-like transition (esMT) that converts compact, adherent ESCs into mesenchymal-like spindle-shaped, highly invasive and metastatic pluripotent stem cells.⁴ This fully reversible process resembles the epithelial-to-mesenchymal transition (EMT), which is essential for normal development and contributes to pathological cancer progression.^{6, 7, 8} Interestingly, the Aldh18a1 gene is specifically induced in and marks the Primitive Endoderm (PrE) in the time window when the pluripotent epiblast precursors are specified within the inner cell mass (ICM) of the blastocyst.⁹ Since the Aldh18a1 enzyme catalyses the first and rate-limiting step of L-Pro biosynthesis, these findings suggest that L-Pro metabolism may regulate cell lineage segregation in early mammalian embryos. Despite its relevance, the molecular mechanisms underlying L-Pro control of stem cell identity remain largely unknown. This prompted us to investigate the early molecular events regulated by exogenously provided L-Pro in mouse ESCs.     

Development of ligands at γ-aminobutyrric acid type A (GABAA) receptor subtype as new agents for pain relief

The identification of compounds with selective anxiolytic-like effects, exerted through the benzodiazepine site on γ-aminobutyric acid type A (GABA_A) receptors, and that show pronounced antihyperalgesia in several pain models, has oriented research towards the development of new agents for the relief of pain. Starting from our previously reported ligands at the benzodiazepine site on GABA_A receptors showing selective anxiolytic-like effects, we have designed new compounds with the aim of identifying those devoid of the typical side effects of the classical benzodiazepines. Our preliminary results indicate that compounds 4, 10(±) and 11 have a very promising antihyperalgesic profile in different animal pain models (peripheral mono-neuropathy, STZ-induced hyperalgesia). In particular 11 exhibits high potency since it exerted its protective effect starting from the dose of 3 mg/kg po, after single injection.     

An aggregation framework to link indicators associated with multifunctional land use to the stakeholder evaluation of policy options

In the last decade efforts have been carried out by the scientific community aimed at building integrated frameworks to support the decision-making process when sustainability issues are addressed. This paper proposes a further advancement in integrated assessment procedures by setting up an operational multi-scale and transparent framework, which comprises the assessment of European regions in terms of sustainability, and the identification of the impact that policy options might have on the sustainability of these regions. The framework is designed for use in ex ante sustainability impact assessment of policy scenarios on multifunctionality of land use and integrates economic, environmental and social issues across a variety of sectors (agriculture, forestry, transport, tourism and energy). The proposed method provides a conceptual framework applicable at different scales (European, regional), and takes into account the great variability of European regions. The described methodology is based on linear additive models to weight and aggregate selected indicators to a set of land use functions identified to describe the goods and services provided by the different land uses that summarise the most relevant economic, environmental and social issues of a region. The framework is designed to allow the evaluation of impacts at an international scale (e.g. the European Union), or on selected regions.The aggregation framework can be used to evaluate the impact that policy options have on the sustainability of multifunctional land use systems with competing demands. A conceptual envelope, called the “trade-off evaluation space”, delineates all possible developments in the functions of the land. The sustainability limits identify the subset of ‘acceptable’ policy options within the trade-off evaluation space, so that the distance of each land use function from sustainability limits can be estimated and trade-offs between the different functions of the multifunctional land use system can be identified. The proposed methodology is adaptable to different contexts: if the assumption is taken that all land use functions are equally weighted the framework can be used to analyse policy cases and take decisions on policy options at the European or regional level. However, at the local-scale the framework can also be applied through a participatory approach and the distribution of weights can be rediscussed with local stakeholders. In both cases the proposed system can be used as a tool for discussion among all interested parties.     

Estimation de la distensibilité aortique à partir de séquences d’images de résonance magnétique « steady-state free-precession » et « contraste de phase »

Objective

To investigate the accuracy in the estimation of the aortic lumen area and distensibility obtained from Steady-State Free-Precession (SSFP) and Phase Contrast (PC) MR sequences.

Subjects and methods

Systolic and diastolic aortic lumen of the ascending and descending aorta of 50 subjects, collected with both acquisition sequences, were extracted using a 2D + t automated segmentation method.

Results

While the statistical study indicated very similar lumen areas for SSFP and PC data (slope = 1, r = 0.99), the corresponding distensibility values were less correlated (r = 0.54). Comparison between distensibility values in the ascending and descending aorta and study of these values with aging indicated a greater coherence with expected physiological behavior of the aorta when using SSFP images.

Discussion

Flow artifacts were more severe in PC images and could explain the better results obtained when using SSFP sequences.

Conclusion

A more accurate local assessment of the aortic stiffness is obtained from SSFP image sequences than from PC image sequences.