Listeria monocytogenes induces host DNA damage and delays the host cell cycle to promote infection

Listeria monocytogenes (Lm) is a human intracellular pathogen widely used to uncover the mechanisms evolved by pathogens to establish infection. However, its capacity to perturb the host cell cycle was never reported. We show that Lm infection affects the host cell cycle progression, increasing its overall duration but allowing consecutive rounds of division. A complete Lm infectious cycle induces a S-phase delay accompanied by a slower rate of DNA synthesis and increased levels of host DNA strand breaks. Additionally, DNA damage/replication checkpoint responses are triggered in an Lm dose-dependent manner through the phosphorylation of DNA-PK, H2A.X, and CDC25A and independently from ATM/ATR. While host DNA damage induced exogenously favors Lm dissemination, the override of checkpoint pathways limits infection. We propose that host DNA replication disturbed by Lm infection culminates in DNA strand breaks, triggering DNA damage/replication responses, and ensuring a cell cycle delay that favors Lm propagation.     

A spontaneous mutation in the nicotinamide nucleotide transhydrogenase gene of C57BL/6J mice results in mitochondrial redox abnormalities

NADPH is the reducing agent for mitochondrial H₂O₂ detoxification systems. Nicotinamide nucleotide transhydrogenase (NNT), an integral protein located in the inner mitochondrial membrane, contributes to an elevated mitochondrial NADPH/NADP⁺ ratio. This enzyme catalyzes the reduction of NADP⁺ at the expense of NADH oxidation and H⁺ reentry to the mitochondrial matrix. A spontaneous Nnt mutation in C57BL/6J (B6J-Nnt^MUT) mice arose nearly 3 decades ago but was only discovered in 2005. Here, we characterize the consequences of the Nnt mutation on the mitochondrial redox functions of B6J-Nnt^MUT mice. Liver mitochondria were isolated both from an Nnt wild-type C57BL/6 substrain (B6JUnib-Nnt^W) and from B6J-Nnt^MUT mice. The functional evaluation of respiring mitochondria revealed major redox alterations in B6J-Nnt^MUT mice, including an absence of transhydrogenation between NAD and NADP, higher rates of H₂O₂ release, the spontaneous oxidation of NADPH, the poor ability to metabolize organic peroxide, and a higher susceptibility to undergo Ca²⁺-induced mitochondrial permeability transition. In addition, the mitochondria of B6J-Nnt^MUT mice exhibited increased oxidized/reduced glutathione ratios as compared to B6JUnib-Nnt^W mice. Nonetheless, the maximal activity of NADP-dependent isocitrate dehydrogenase, which is a coexisting source of mitochondrial NADPH, was similar between both groups. Altogether, our data suggest that NNT functions as a high-capacity source of mitochondrial NADPH and that its functional loss due to the Nnt mutation results in mitochondrial redox abnormalities, most notably a poor ability to sustain NADP and glutathione in their reduced states. In light of these alterations, the potential drawbacks of using B6J-Nnt^MUT mice in biomedical research should not be overlooked.     

An integrated network analysis reveals that nitric oxide reductase prevents metabolic cycling of nitric oxide by Pseudomonas aeruginosa

Nitric oxide (NO) is a chemical weapon within the arsenal of immune cells, but is also generated endogenously by different bacteria. Pseudomonas aeruginosa are pathogens that contain an NO-generating nitrite (NO₂⁻) reductase (NirS), and NO has been shown to influence their virulence. Interestingly, P. aeruginosa also contain NO dioxygenase (Fhp) and nitrate (NO₃⁻) reductases, which together with NirS provide the potential for NO to be metabolically cycled (NO→NO₃⁻→NO₂⁻→NO). Deeper understanding of NO metabolism in P. aeruginosa will increase knowledge of its pathogenesis, and computational models have proven to be useful tools for the quantitative dissection of NO biochemical networks. Here we developed such a model for P. aeruginosa and confirmed its predictive accuracy with measurements of NO, O₂, NO₂⁻, and NO₃⁻ in mutant cultures devoid of Fhp or NorCB (NO reductase) activity. Using the model, we assessed whether NO was metabolically cycled in aerobic P. aeruginosa cultures. Calculated fluxes indicated a bottleneck at NO₃⁻, which was relieved upon O₂ depletion. As cell growth depleted dissolved O₂ levels, NO₃⁻ was converted to NO₂⁻ at near-stoichiometric levels, whereas NO₂⁻ consumption did not coincide with NO or NO₃⁻ accumulation. Assimilatory NO₂⁻ reductase (NirBD) or NorCB activity could have prevented NO cycling, and experiments with ΔnirB, ΔnirS, and ΔnorC showed that NorCB was responsible for loss of flux from the cycle. Collectively, this work provides a computational tool to analyze NO metabolism in P. aeruginosa, and establishes that P. aeruginosa use NorCB to prevent metabolic cycling of NO.     

Life cycle ofPseudodiamesa branickii (Chironomidae) in a small upland stream

The generation time ofP. branickii was studied using larval samples in conjunction with rearing experiments and continuous collection of egg masses across one year. This species produced three generations per year in a central German stream (280 m a.s.l., 50° 40 N). Its generation time was variable and obviously influenced by the photoperiod to which eggs and larvulae were subjected. It is thus concluded that two strains ofP. branickii were present in a single population, one bivoltine and the other trivoltine.     

Projecting future grassland productivity to assess the sustainability of potential biofuel feedstock areas in the Greater Platte River Basin

This study projects future (e.g., 2050 and 2099) grassland productivities in the Greater Platte River Basin (GPRB) using ecosystem performance (EP, a surrogate for measuring ecosystem productivity) models and future climate projections. The EP models developed from a previous study were based on the satellite vegetation index, site geophysical and biophysical features, and weather and climate drivers. The future climate data used in this study were derived from the National Center for Atmospheric Research Community Climate System Model 3.0 ‘SRES A1B’ (a ‘middle’ emissions path). The main objective of this study is to assess the future sustainability of the potential biofuel feedstock areas identified in a previous study. Results show that the potential biofuel feedstock areas (the more mesic eastern part of the GPRB) will remain productive (i.e., aboveground grassland biomass productivity >2750 kg ha^?1 year^?1) with a slight increasing trend in the future. The spatially averaged EPs for these areas are 3519, 3432, 3557, 3605, 3752, and 3583 kg ha^?1 year^?1 for current site potential (2000–2008 average), 2020, 2030, 2040, 2050, and 2099, respectively. Therefore, the identified potential biofuel feedstock areas will likely continue to be sustainable for future biofuel development. On the other hand, grasslands identified as having no biofuel potential in the drier western part of the GPRB would be expected to stay unproductive in the future (spatially averaged EPs are 1822, 1691, 1896, 2306, 1994, and 2169 kg ha^?1 year^?1 for site potential, 2020, 2030, 2040, 2050, and 2099). These areas should continue to be unsuitable for biofuel feedstock development in the future. These future grassland productivity estimation maps can help land managers to understand and adapt to the expected changes in future EP in the GPRB and to assess the future sustainability and feasibility of potential biofuel feedstock areas.     

Impact of Urban Pollution from the Hanoi Area on Benthic Diatom Communities Collected from the Red, Nhue and Tolich Rivers (Vietnam)

The effects of urban pollution from Hanoi city on the benthic diatom communities of the Nhue–Tolich river system were studied during the 2003 dry season. Benthic diatoms were allowed to grow on glass slides suspended in the water flow for 4 weeks. To reveal the relationship between water quality and diatom communities, Canonical correspondence analysis (CCA) was used on data concerning relative abundances of diatom species and environmental variables. Two diatom indices, IPS and DAIpo, were applied to evaluate water quality in the three rivers. A total of 291 diatom taxa were found in the Red, Nhue and Tolich Rivers. These were mainly cosmopolitan taxa, with some tropical, subtropical and endemic taxa. The most abundant taxa at the Red site were Aulacoseira granulata, Achnanthidium minutissimum, Encyonema minutum, Navicula recens and other halophilous taxa such as Nitzschia kurzii, Seminavis strigosa, Entomoneis paludosa, Bacillaria paradoxa. Diatom assemblages at the Tolich site consisted mainly of Nitzschia umbonata, Nitzschia palea and Eolimna minima. Diatom density ranged from 660 to 30,000 cells/cm². Environmental variables and diatom assemblage composition at all sites were significantly correlated. Two diatom indices gave similar results and indicate the Tolich River with the lowest values as a highly polluted site.     

Vulnerability assessment of wetland landscape ecosystem services using driver-pressure-state-impact-response (DPSIR) model

Wetlands exist in complex ecological conditions that are changeable in time and space in terms of function and structural diversity. In recent decades, wetlands have been exposed to a wide range of threats. Assessment of these threats is essential to develop an understanding of the state of a wetland ecosystem and to develop a suitable management strategy. This paper discusses wetland vulnerability in terms of analysis of human and environmental systems from application of the driver-pressure-state-impact-response (DPSIR) framework. This assessment presented a systematic methodology for assessment of wetland vulnerability in a social-ecological approach applying broad-scale ecosystem services and vulnerability functions. The method combined the hydro-geomorphic approach with estimations of vulnerability indicators and DPSIR analysis. The aim of this paper was to assess vulnerability of wetland ecosystem services and to characterize the threat indicators according to importance, severity, and probability of occurrence. Quantitative and qualitative methods were applied to characterize values for these three indicators. The Multi Criteria Decision Making (MCDM) method was used to prioritize threats and impacts of the wetland on the basis of experts’ opinions. The proposed methodology was applied to the Choghakhor international wetland landscape in south-western Iran. Vulnerability assessment revealed that water requirement of the lowland and the water transfer system were the most important factors threatening the wetland. Agricultural activities, settlements and urban areas, drought, tourism, population growth, and mining activities in the upland were the next most important priorities, in that order. Hydrological balance was determined as the most vulnerable function and was considered as the most important function in the Choghakhor wetland. The DPSIR model was used to determine a management strategy to reduce vulnerability of ecosystem services in response to drivers, pressures, states and impacts indicated by modelling.     

A monoclonal antibody specific for prophase phosphorylation of histone deacetylase 1: a readout for early mitotic cells

Histone deacetylases (HDACs) are modification enzymes that regulate a plethora of biological processes. HDAC1, a crucial epigenetic modifier, is deregulated in cancer and subjected to a variety of post-translational modifications. Here, we describe the generation of a new monoclonal antibody that specifically recognizes a novel highly dynamic prophase phosphorylation of serine 406-HDAC1, providing a powerful tool for detecting early mitotic cells.