Pachybrachis sassii, a new species from the Mediterranean Giglio Island (Italy) (Coleoptera, Chrysomelidae, Cryptocephalinae)

Pachybrachis sassii, new species is described from Giglio Island, of the Tuscan Archipelago (Italy). The new species belongs to the nominotypical subgenus and is closely related to Pachybrachis salfii Burlini, 1957, from which it differs in the shape of the median lobe of the aedeagus and in the pattern of the yellow raised spots on the elytra and pronotum. Ecological observations are made. The neotype of Pachybrachis salfii from Colloreto, Monte Pollino (Italy) is designated.     

Single-cell analysis of ploidy and centrosomes underscores the peculiarity of normal hepatocytes

Polyploidization is the most well recognized feature of the liver. Yet, a quantitative and behavioral analysis of centrosomes and DNA content in normal hepatocytes has been limited by the technical challenges of methods available. By using a novel approach employing FISH for chromosomes 18, X and Y we provide, for the first time, a detailed analysis of DNA copies during physiological development in the liver at single cell level. We demonstrate that aneuploidy and unbalanced DNA content in binucleated hepatocytes are common features in normal adult liver. Despite the common belief that hepatocytes contain 1, 2 or no more than 4 centrosomes, our double staining for centrosome associated proteins reveals extranumerary centrosomes in a high percentage of cells as early as 15 days of age. We show that in murine liver the period between 15 days and 1.5 months marks the transition from a prevalence of mononucleated cells to up to 75% of binucleated cells. Our data demonstrate that this timing correlates with a switch in centrosomes number. At 15 days the expected 1 or 2 centrosomes converge with several hepatocytes that contain 3 centrosomes; at 1.5 months the percentage of cells with 3 centrosomes decreases concomitantly with the increase of cells with more than 4 centrosomes. Our analysis shows that the extranumerary centrosomes emerge in concomitance with the process of binucleation and polyploidization and maintain α-tubulin nucleation activity. Finally, by integrating interphase FISH and immunofluorescent approaches, we detected an imbalance between centrosome number and DNA content in liver cells that deviates from the equilibrium expected in normal cells. We speculate that these unique features are relevant to the peculiar biological function of liver cells which are continuously challenged by stress, a condition that could predispose to genomic instability.     

Spatial and bathymetric trends in Harpacticoida (Copepoda) community structure in the Northern Gulf of Mexico deep-sea

Harpacticoid copepod community structure was analyzed at 43 stations in the northern Gulf of Mexico deep-sea to test regional and bathymetric patterns of diversity in relation to environmental variables and topographic complexity of the continental slope. Depth, longitude, and proximity to the Florida Escarpment significantly affect average phylogenetic diversity, but basins and canyons do not. Multivariate analysis reveals a significant inverse relationship between diversity and POM flux, which is confirmed by significant region-scale depth and longitude differences. Although species richness declines linearly with increasing depth, the expected number of species (rarefraction) is maximized at approximately 1200 m, and average taxonomic and phylogenetic diversity continue to increase with depth, suggesting greater morphological or functional harpacticoid diversity with increasing depth. Most stations have unique species compositions, suggesting high regional (2200 species) and global (10⁵-10⁶ species) diversity by extrapolation. Therefore, processes maintaining harpacticoid diversity in the northern Gulf of Mexico deep-sea seem to rely on both small-scale dispersal and large-scale food supply mechanisms.     

RAD51 Mutants Cause Replication Defects and Chromosomal Instability

RAD51 is important for restarting stalled replication forks and for repairing DNA double-strand breaks (DSBs) through a pathway called homology-directed repair (HDR). However, analysis of the consequences of specific RAD51 mutants has been difficult since they are toxic. Here we report on the dominant effects of two human RAD51 mutants defective for ATP binding (K133A) or ATP hydrolysis (K133R) expressed in mouse embryonic stem (ES) cells that also expressed normal mouse RAD51 from the other chromosome. These cells were defective for restarting stalled replication forks and repairing breaks. They were also hypersensitive to camptothecin, a genotoxin that generates breaks specifically at the replication fork. In addition, these cells exhibited a wide range of structural chromosomal changes that included multiple breakpoints within the same chromosome. Thus, ATP binding and hydrolysis are essential for chromosomal maintenance. Fusion of RAD51 to a fluorescent tag (enhanced green fluorescent protein [eGFP]) allowed visualization of these proteins at sites of replication and repair. We found very low levels of mutant protein present at these sites compared to normal protein, suggesting that low levels of mutant protein were sufficient for disruption of RAD51 activity and generation of chromosomal rearrangements.     

A restoration suitability index model for the eastern oyster (Crassostrea virginica) in the Mission-Aransas Estuary, TX, USA

Oyster reefs are one of the most threatened marine habitats on earth, with habitat loss resulting from water quality degradation, coastal development, destructive fishing practices, overfishing, and storm impacts. For successful and sustainable oyster reef restoration efforts, it is necessary to choose sites that support long-term growth and survival of oysters. Selection of suitable sites is critically important as it can greatly influence mortality factors and may largely determine the ultimate success of the restoration project. The application of Geographic Information Systems (GIS) provides an effective methodology for identifying suitable sites for oyster reef restoration and removes much of the uncertainty involved in the sometimes trial and error selection process. This approach also provides an objective and quantitative tool for planning future oyster reef restoration efforts. The aim of this study was to develop a restoration suitability index model and reef quality index model to characterize locations based on their potential for successful reef restoration within the Mission-Aransas Estuary, Texas, USA. The restoration suitability index model focuses on salinity, temperature, turbidity, dissolved oxygen, and depth, while the reef quality index model focuses on abundance of live oysters, dead shell, and spat. Size-specific Perkinsus marinus infection levels were mapped to illustrate general disease trends. This application was effective in identifying suitable sites for oyster reef restoration, is flexible in its use, and provides a mechanism for considering alternative approaches. The end product is a practical decision-support tool that can be used by coastal resource managers to improve oyster restoration efforts. As oyster reef restoration activities continue at small and large-scales, site selection criteria are critical for assisting stakeholders and managers and for maximizing long-term sustainability of oyster resources.     

AMPKα loss promotes KRAS-mediated lung tumorigenesis

AMP-activated protein kinase (AMPK) is a critical sensor of energy status that coordinates cell growth with energy balance. In non-small cell lung cancer (NSCLC) the role of AMPKα is controversial and its contribution to lung carcinogenesis is not well-defined. Furthermore, it remains largely unknown whether long non-coding RNAs (lncRNAs) are involved in the regulation of AMPK-mediated pathways. Here, we found that loss of AMPKα in combination with activation of mutant KRAS^G12D increased lung tumour burden and reduced survival in Kras^LSLG12D/+/AMPKα^fl/fl mice. In agreement, functional in vitro studies revealed that AMPKα silencing increased growth and migration of NSCLC cells. In addition, we identified an AMPKα-modulated lncRNA, KIMAT1 (ENSG00000228709), which in turn regulates AMPKα activation by stabilizing the lactate dehydrogenase B (LDHB). Collectively, our study indicates that AMPKα loss promotes KRAS-mediated lung tumorigenesis and proposes a novel KRAS/KIMAT1/LDHB/AMPKα axis that could be exploited for therapeutic purposes.Subject terms: Cancer models, Non-small-cell lung cancer     

Role and Value of Nitrogen Regulation Provided by Oysters (Crassostrea virginica) in the Mission-Aransas Estuary,Texas, USA

Suspension-feeding activities of oysters impart a potentially significant benefit to estuarine ecosystems via reduction of water column nutrients, plankton and seston biomass, and primary productivity which can have a significant impact on human well-being. This study considered nitrogen regulation by eastern oysters Crassostrea virginica in the Mission-Aransas Estuary, Texas, USA, as a function of denitrification, burial, and physical transport from the system via harvest. Oyster reefs were estimated to remove 502.5 kg N km⁻² through denitrification of biodeposits and 251.3 kg N km⁻² in burial of biodeposits to sediments. Nitrogen is also physically transported out of the estuary via harvest of oysters. Commercial harvest of oysters in the Mission-Aransas Estuary can remove approximately 21,665 kg N per year via physical transport from the system. We developed a transferable method to value the service of nitrogen regulation by oysters, where the potential cost equivalent value of nitrogen regulation is quantified via cost estimates for a constructed biological nutrient removal (BNR) supplement to a wastewater treatment plant. The potential annual engineered cost equivalent of the service of nitrogen regulation and removal provided by reefs in the Mission-Aransas Estuary is $293,993 yr⁻¹. Monetizing ecosystem services can help increase awareness at the stakeholder level of the importance of oysters beyond commercial fishery values alone.