Loss of GdpP function in Staphylococcus aureus leads to β-lactam tolerance and enhanced evolution of β-lactam resistance

Infections caused by Staphylococcus aureus are a leading cause of mortality. Treating infections caused by S. aureus is difficult due to resistance against most traditional antibiotics, including β-lactams. We previously reported the presence of mutations in gdpP among S. aureus strains that were obtained by serial passaging in β-lactam drugs. Similar mutations have recently been reported in natural S. aureus isolates that are either non-susceptible or resistant to β-lactam antibiotics. gdpP codes for a phosphodiesterase that cleaves cyclic-di-AMP (CDA), a newly discovered second messenger. In this study, we sought to identify the role of gdpP in β-lactam resistance in S. aureus . Our results showed that gdpP associated mutations caused loss of phosphodiesterase function, leading to increased CDA accumulation in the bacterial cytosol. Deletion of gdpP led to an enhanced ability of the bacteria to withstand a β-lactam challenge (two to three log increase in bacterial colony forming units) by promoting tolerance without enhancing MICs of β-lactam antibiotics. Our results demonstrated that increased drug tolerance due to loss of GdpP function can provide a selective advantage in acquisition of high-level β-lactam resistance. Loss of GdpP function thus increases tolerance to β-lactams that can lead to its therapy failure and can permit β-lactam resistance to occur more readily.

Proposal of a New Method for Controlling the Thaw of Permafrost around the China–Russia Crude Oil Pipeline and a Preliminary Study of Its Ventilation Capacity

The China–Russia crude oil pipeline (CRCOP) has been in operation for over ten years. Field observation results have shown that a thaw bulb has developed around the CRCOP which expands at a rate of more than 0.8 m∙a−1 in depth. In view of the deficits of existing measures in mitigating permafrost thaw, a new control method is proposed based on active cooling. According to the relationship between total pressure loss and the driving force of natural ventilation, the wind speed in a U-shaped air-ventilation pipe around the CRCOP is calculated. By analyzing the theoretical calculation and numerical analysis results, it is found that the influence of thermal pressure difference on the natural ventilation of the structure can be negligible, and the influences of resistance loss along the pipe and local resistance loss in the pipe are similarly negligible. Exhaust elbows greatly improve the ventilation performance of the U-shaped air-ventilated pipe. This study developed a novel structure around warm-oil pipelines in permafrost for mitigating thaw settlement along the CRCOP and other similar projects across the world.

Rheological Parameters and Transport Characteristics of Fresh Cement Tailings Backfill Slurry in an Underground Iron Mine

Pipeline transportation is the key component of the mine filling system. In this study, fresh cement tailing backfill (CTB) slurry made by unclassified tailings from the Daye iron mine is taken as the research object, and its rheological parameters and transport characteristics are studied via laboratory test and FLUENT software. It was found that the relationship curve of the dynamic yield stress, viscosity, and solid content (SC) of CTB slurry fits the law of the H-B model when SC varies between 60% and 68%. However, the relationship curve gradually changes to fit the Bingham mode when SC reaches up to 70%. Numerical simulation results demonstrate that when the SC of CTB slurry exceeds 65%, the static pressure at the pipeline’s outlet begins to distribute symmetrically. At this point, the slurry flow state is relatively stable, and the pipeline resistance loss is positively correlated with SC and flow rate. When SC exceeds 70%, resistance loss begins to increase significantly. The findings of this study can be used to identify the suitable transportation conditions of CTB slurry and provide the theoretical direction for the pipeline transportation design of filling systems in mines.

Influence of Solid Content, Cement/Tailings Ratio, and Curing Time on Rheology and Strength of Cemented Tailings Backfill

Understanding the flow process of cemented tailings backfill (CTB) is important for successful pumping into underground stopes. This study examines the effects of solid content (SC), cement/tailings (c/t) ratio, and curing time (CT) on rheological and mechanical properties of CTB mixes. The slurry concentration of the mixes was 65, 67, and 69 wt. %, with c/t ratios ranging from 1:4 to 1:20. Unconfined compressive strength (UCS) tests were performed on hardened CTB mixes after curing 3, 7, and 28 days. The rheological properties of CTB slurries are mainly related to SC. The yield stress and viscosity of fresh mixes increase with increasing SC, but the pipeline resistance loss (PRL) also increases with increasing SC. According to the analysis of variance, the SC and flow rate are the most significant parameters which greatly affect the PRL performance. The c/t and CT parameters are the most significant parameters for affecting the shrinkage rate. The findings offer a reference for theoretical optimization for mine filling systems of similar type.

The MEK/ERK Module Is Reprogrammed in Remodeling Adult Cardiomyocytes

Fetal and hypertrophic remodeling are hallmarks of cardiac restructuring leading chronically to heart failure. Since the Ras/Raf/MEK/ERK cascade (MAPK) is involved in the development of heart failure, we hypothesized, first, that fetal remodeling is different from hypertrophy and, second, that remodeling of the MAPK occurs. To test our hypothesis, we analyzed models of cultured adult rat cardiomyocytes as well as investigated myocytes in the failing human myocardium by western blot and confocal microscopy. Fetal remodeling was induced through endothelial morphogens and monitored by the reexpression of Acta2, Actn1, and Actb. Serum-induced hypertrophy was determined by increased surface size and protein content of cardiomyocytes. Serum and morphogens caused reprogramming of Ras/Raf/MEK/ERK. In both models H-Ras, N-Ras, Rap2, B- and C-Raf, MEK1/2 as well as ERK1/2 increased while K-Ras was downregulated. Atrophy, MAPK-dependent ischemic resistance, loss of A-Raf, and reexpression of Rap1 and Erk3 highlighted fetal remodeling, while A-Raf accumulation marked hypertrophy. The knock-down of B-Raf by siRNA reduced MAPK activation and fetal reprogramming. In conclusion, we demonstrate that fetal and hypertrophic remodeling are independent processes and involve reprogramming of the MAPK.

Structural basis of nanobody recognition of grapevine fanleaf virus and of virus resistance loss

Grapevine fanleaf virus (GFLV) is a picorna-like plant virus transmitted by nematodes that affects vineyards worldwide. Nanobody (Nb)-mediated resistance against GFLV has been created recently, and shown to be highly effective in plants, including grapevine, but the underlying mechanism is unknown. Here we present the high-resolution cryo electron microscopy structure of the GFLV–Nb23 complex, which provides the basis for molecular recognition by the Nb. The structure reveals a composite binding site bridging over three domains of one capsid protein (CP) monomer. The structure provides a precise mapping of the Nb23 epitope on the GFLV capsid in which the antigen loop is accommodated through an induced-fit mechanism. Moreover, we uncover and characterize several resistance-breaking GFLV isolates with amino acids mapping within this epitope, including C-terminal extensions of the CP, which would sterically interfere with Nb binding. Escape variants with such extended CP fail to be transmitted by nematodes linking Nb-mediated resistance to vector transmission. Together, these data provide insights into the molecular mechanism of Nb23-mediated recognition of GFLV and of virus resistance loss.