Balancing crosstalk between 20-hydroxyecdysone-induced autophagy and caspase activity in the fat body during Drosophila larval-prepupal transition

In the fruitfly, Drosophila melanogaster, autophagy and caspase activity function in parallel in the salivary gland during metamorphosis and in a common regulatory hierarchy during oogenesis. Both autophagy and caspase activity progressively increase in the remodeling fat body, and they are induced by a pulse of the molting hormone (20-hydroxyecdysone, 20E) during the larval-prepupal transition. Inhibition of autophagy and/or caspase activity in the remodeling fat body results in 25–40% pupal lethality, depending on the genotypes. Interestingly, a balancing crosstalk occurs between autophagy and caspase activity in this tissue: the inhibition of autophagy induces caspase activity and the inhibition of caspases induces autophagy. The Drosophila remodeling fat body provides an in vivo model for understanding the molecular mechanism of the balancing crosstalk between autophagy and caspase activity, which oppose with each other and are induced by the common stimulus 20E, and blockage of either path reinforces the other path.     

Investigating the role of active site residues of Rhodotorula gracilis D-amino acid oxidase on its substrate specificity

D-amino acid oxidase (DAAO) is a flavoprotein that catalyzes stereospecifically the oxidative deamination of D-amino acids. The wild-type DAAO is mainly active on neutral D-amino acids, while basic D-amino acids are poor substrates and the acidic ones are virtually not oxidized. To present a comprehensive picture of how the active site residues can modulate the substrate specificity a number of mutants at position M213, Y223, Y238, R285, S335, and Q339 were prepared in the enzyme from the yeast Rhodotorula gracilis. All DAAO mutants have spectral properties similar to those of the wild-type enzyme and are catalytically active, thus excluding an essential role in catalysis; a lower activity on neutral and basic amino acids was observed. Interestingly, an increase in activity and (k(cat)/K(m))(app) ratio on D-aspartate was observed for all the mutants containing an additional charged residue in the active site. The active site of yeast DAAO appears to be a highly evolved scaffold built up through evolution to optimize the oxidative deamination of neutral D-amino acids without limiting its substrate specificity. It is noteworthy, that introduction of a sole, additional, positively charged residue in the active site is sufficient to optimize the reactivity on acidic D-amino acids, giving rise to kinetic properties similar to those of D-aspartate oxidase.     

New biotech applications from evolved D-amino acid oxidases

D-Amino acid oxidase (DAAO) is a well-known flavoenzyme that catalyzes the oxygen-dependent oxidative deamination of amino acid D-isomers with absolute stereospecificity, which results in α-keto acids, ammonia and hydrogen peroxide. Recently, the extraordinary functional plasticity of DAAO has become evident; in turn, boosting research on this flavoprotein. Protein engineering has allowed for a redesign of DAAO substrate specificity, oxygen affinity, cofactor binding, stability, and oligomeric state. We review recent developments in utilizing DAAO, including as a biocatalyst for resolving racemic amino acid mixtures, as a tool for biosensing, and as a new mechanism of herbicide resistance. Perspectives for future biotechnological applications of this oxidative biocatalyst are also outlined.     

Monitoring tropical forests under a functional perspective with satellite‐based vegetation optical depth

Monitoring ecosystem functions in forests is a priority in a climate change scenario, as climate‐induced events may initially alter the functions more than slow‐changing attributes, such as biomass. The ecosystem functional properties (EFPs) are quantities that characterize key ecosystem processes. They can be derived by point observations of gas and energy exchanges between the ecosystems and the atmosphere that are collected globally at FLUXNET flux tower sites and upscaled at ecosystem level. The properties here considered describe the ability of ecosystems to optimize the use of resources for carbon uptake. They represent functional forest information, are dependent on environmental drivers, linked to leaf traits and forest structure, and influenced by climate change effects. The ability of vegetation optical depth (VOD) to provide forest functional information is investigated using 2011–2014 satellite data collected by the Soil Moisture and Ocean Salinity mission and using the EFPs as reference dataset. Tropical forests in Africa and South America were analyzed, also according to ecological homogeneous units. VOD jointly with water deficit information explained 93% and 87% of the yearly variability in both flux upscaled maximum gross primary productivity and light use efficiency functional properties, in Africa and South America forests respectively. Maps of the retrieved properties evidenced changes in forest functional responses linked to anomalous climate‐induced events during the study period. The findings indicate that VOD can support the flux upscaling process in the tropical range, affected by high uncertainty, and the detection of forest anomalous functional responses. Preliminary temporal analysis of VOD and EFP signals showed fine‐grained variability in periodicity, in signal dephasing, and in the strength of the relationships. In selected drier forest types, these satellite data could also support the monitoring of functional dynamics.     

Modelling the potential spread of Hyalomma marginatum ticks in Europe by migratory birds

This study modelled the probability of introduction of Hyalomma marginatum into Europe by predicting the potential migratory routes of 28 bird species and the probability to carry immatures of the tick. Flyways were modelled as a spatio-temporal feature, at weekly intervals, using satellite-derived data of temperature and vegetal phenology, together with cost surfaces derived from speed and direction of the wind (years 2002–2018). The expected period of activity of tick immatures defined the probability of ticks being carried by birds along the modelled flyways. The probability of moulting of the engorged nymphs was modelled as a linear relationship of the daily sum of temperatures after tick introduction by birds. Positive probabilities of tick introduction extend the known northern range of permanent populations to central and western France, and large portions of central Europe. The flight of birds into an area and thence the risk of introduction of H. marginatum is very heterogeneous, with sites receiving “waves” of different bird species at diverse times of the year. Therefore, there is not a clear period of time for introduction, as it depends on the modelled behaviour of the bird species. The probability of introduction into Baltic and Nordic countries is small. We hypothesise that conditions of a warmer climate might support permanent populations of H. marginatum if a high number of immatures is introduced. Active surveys in risky territories, where the tick is not yet established, are advisable for rapid intervention.     

Dinosaur Tracks as Paleogeographic Constraints: New Scenarios for the Cretaceous Geography of the Periadriatic Region

A really unexpected finding of sauropod and theropod footprints in southern Latium raises to four the number of the trampled levels recognized in central and southern Italy. After the recent findings in Latest Jurassic and Early, mid and Late Cretaceous carbonate platform deposits of the Periadriatic region, dinosaur footprints seem to provide very important paleogeographic constraints for reconstructing the geodynamic history of the Mediterranean area. The presence of a varied ichnoassociation makes acceptance of the current paleogeographic models concerning the relative and absolute position of the Laziale-Abruzzese-Campano and of Apulian-Dinaric domains during the Late Cretaceous more and more problematic. Dinosaur footprints, combined with other paleontological data, demonstrate that these areas were never completely pulled apart by deep seaways, while frequent or continuous links between them, and to southern and northern mainlands, probably persisted. These data also allowed us to improve our understanding of the timing of the Mesozoic plate motion in this segment of the Western Tethys.     

Direct inhibition of hexokinase activity by metformin at least partially impairs glucose metabolism and tumor growth in experimental breast cancer

Emerging evidence suggests that metformin, a widely used anti-diabetic drug, may be useful in the prevention and treatment of different cancers. In the present study, we demonstrate that metformin directly inhibits the enzymatic function of hexokinase (HK) I and II in a cell line of triple-negative breast cancer (MDA-MB-231). The inhibition is selective for these isoforms, as documented by experiments with purified HK I and II as well as with cell lysates. Measurements of ¹⁸F-fluoro-deoxyglycose uptake document that it is dose- and time-dependent and powerful enough to virtually abolish glucose consumption despite unchanged availability of membrane glucose transporters. The profound energetic imbalance activates phosphorylation and is subsequently followed by cell death. More importantly, the “in vivo” relevance of this effect is confirmed by studies of orthotopic xenografts of MDA-MB-231 cells in athymic (nu/nu) mice. Administration of high drug doses after tumor development caused an evident tumor necrosis in a time as short as 48 h. On the other hand, 1 mo metformin treatment markedly reduced cancer glucose consumption and growth. Taken together, our results strongly suggest that HK inhibition contributes to metformin therapeutic and preventive potential in breast cancer.     

EV20, a Novel Anti-ErbB-3 Humanized Antibody,Promotes ErbB-3 Down-Regulation and Inhibits Tumor Growth In Vivo

ErbB-3 (HER-3) receptor is involved in tumor progression and resistance to therapy. Development of specific inhibitors impairing the activity of ErbB-3 is an attractive tool for cancer therapeutics. MP-RM-1, a murine monoclonal antibody targeting human ErbB-3, has shown anticancer activity in preclinical models. With the aim to provide novel candidates for clinical use, we have successfully generated a humanized version of MP-RM-1. The humanized antibody, named EV20, abrogates both ligand-dependent and ligand-independent receptor signaling of several tumor cell types, strongly promotes ErbB-3 down-regulation, and efficiently and rapidly internalizes into tumor cells. Furthermore, treatment with EV20 significantly inhibits growth of xenografts originating from prostatic, ovarian, and pancreatic cancers as well as melanoma in nude mice. In conclusion, we provide a novel candidate for ErbB-3-targeted cancer therapy.