Unbalanced Charge Distribution as a Determinant for Dependence of a Subset of Escherichia coli Membrane Proteins on the Membrane Insertase YidC

Membrane proteins are involved in numerous essential cell processes, including transport, gene regulation, motility, and metabolism. To function properly, they must be inserted into the membrane and folded correctly. YidC, an essential protein in Escherichia coli with homologues in other bacteria, Archaea, mitochondria, and chloroplasts, functions by incompletely understood mechanisms in the insertion and folding of certain membrane proteins. Using a genome-scale approach, we identified 69 E. coli membrane proteins that, in the absence of YidC, exhibited aberrant localization by microscopy. Further examination of a subset revealed biochemical defects in membrane insertion in the absence of YidC, indicating their dependence on YidC for proper membrane insertion or folding. Membrane proteins possessing an unfavorable distribution of positively charged residues were significantly more likely to depend on YidC for membrane insertion. Correcting the charge distribution of a charge-unbalanced YidC-dependent membrane protein abrogated its requirement for YidC, while perturbing the charge distribution of a charge-balanced YidC-independent membrane protein rendered it YidC dependent, demonstrating that charge distribution can be a necessary and sufficient determinant of YidC dependence. These findings provide insights into a mechanism by which YidC promotes proper membrane protein biogenesis and suggest a critical function of YidC in all organisms and organelles that express it

The MH1 domain of Smad3 interacts with Pax6 and represses autoregulation of the Pax6 P1 promoter

Pax6 transcription is under the control of two main promoters (P0 and P1), and these are autoregulated by Pax6. Additionally, Pax6 expression is under the control of the TGFβ superfamily, although the precise mechanisms of such regulation are not understood. The effect of TGFβ on Pax6 expression was studied in the FHL124 lens epithelial cell line and was found to cause up to a 50% reduction in Pax6 mRNA levels within 24 h. Analysis of luciferase reporters showed that Pax6 autoregulation of the P1 promoter, and its induction of a synthetic promoter encoding six paired domain-binding sites, were significantly repressed by both an activated TGFβ receptor and TGFβ ligand stimulation. Subsequently, a novel Pax6 binding site in P1 was shown to be necessary for autoregulation, indicating a direct influence of Pax6 protein on P1. In transfected cells, and endogenously in FHL124 cells, Pax6 co-immunoprecipitated with Smad3 following TGFβ receptor activation, while in GST pull-down experiments, the MH1 domain of Smad3 was observed binding the RED sub-domain of the Pax6 paired domain. Finally, in DNA adsorption assays, activated Smad3 inhibited Pax6 from binding the consensus paired domain recognition sequence. We hypothesize that the Pax6 autoregulatory loop is targeted for repression by the TGFβ/Smad pathway, and conclude that this involves diminished paired domain DNA-binding function resulting from a ligand-dependant interaction between Pax6 and Smad3

Catalytic meta-selective C–H functionalization to construct quaternary carbon centres

A catalytic meta-selective C-H functionalization of 2-phenylpyridines using a range of tertiary halides is described. The protocol is simple to perform and usees commercially available reagents to construct challenging quaternary carbon centres in a regioselective manner. Preliminary studies suggest the C-H functionalization proceeds trhough a radical process directed via a remote sigma activation

Remote sensing of vegetation fires and its contribution to a fire management information system

In the last decade, research has proven that remote sensing can provide very useful support to fire managers. This chapter provides an overview of the type of information remote sensing can provide to the fire community. It considers first fire management information needs in the context of fire management information system. An introduction to remote sensing then precedes the description of fire information obtainable from remote sensing data (such as vegetation status, active fire detection and burned areas assessment). Finally, operational examples in five African countries illustrate how the information can be used in practice

Isothiourea-catalyzed enantioselective addition of 4-nitrophenyl esters to iminium ions

The authors thank AstraZeneca and the EPSRC (grant codes EP/M506631/1; J.N.A. and EP/J018139/1; A.B.F.) for funding. The European Research Council under the European Union’s Seventh Framework Programme (FP7/2007-2013) ERC Grant Agreement No. 279850 is also acknowledged. A.D.S. thanks the Royal Society for a Wolfson Research Merit Award.Isothioureas catalyze the enantioselective addition of 4-nitrophenyl esters to tetrahydroisoquinoline-derived iminium ions. 4-Nitrophenoxide, generated in situ from initial N-acylation of the isothiourea by the 4-nitrophenyl ester, is used to facilitate catalyst turnover in this reaction process. Optimization showed that 4-nitrophenyl esters give the best reactivity in this protocol over a range of alternative aryl esters, with the observed enantioselectivity markedly dependent upon the nature of the iminium counteri-on. Highest yields and enantioselectivity were obtained using iminium bromide ions generated in situ via photoredox catalysis using BrCCl3 and Ru(bpy)3Cl2 (0.5 mol%) and commercially available tetramisole (5 mol%) as the Lewis base catalyst. The scope and limitations of this procedure was developed, giving the desired β-amino amide products in up to 96% yield, 79:21 dr and ermajor (2R,1′S) 99.5:0.5.PostprintPeer reviewe

8. Remote Sensing Of Vegetation Fires And Its Contribution To A Fire Management Information System

In the last decade, research has proven that remote sensing can provide very useful support to fire managers. This chapter provides an overview of the types of information remote sensing can provide to the fire community. First, it considers fire management information needs in the context of a fire management information system. An introduction to remote sensing then precedes a description of fire information obtainable from remote sensing data (such as vegetation status, active fire detection and burned areas assessment). Finally, operational examples in five African countries illustrate the practical use of remotely sensed fire information. As indicated in previous chapters, fire management usually comprises activities designed to control the frequency, area, intensity or impact of fire. These activities are undertaken in different institutional, economic, social, environmental and geographical contexts, as well as at different scales, from local to national. The range of fire management activities also varies considerably according to the management issues at stake, as well as the available means and capacity to act. Whatever the level, effective fire management requires reliable information upon which to base appropriate decisions and actions. Information will be required at many different stages of this fire management system. To illustrate this, we consider a typical and generic description of a fire management loop , as provided in Figure 8.1. Fire management objectives result from fire related knowledge . For example, they may relate to sound ecological reasons for prescribed burning in a particular land management context, or to frequent, uncontrolled fires threatening valuable natural or human resources. Whatever the issues, appropriate objectives require scientific knowledge (such as fire impact on ecosystems components, such as soil and vegetation), as well as up-to date monitoring information (such as vegetation status, fire locations, land use, socioeconomic context, etc.). Policies, generally at a national and governmental level, provide the official or legal long term framework (e.g. five to ten years) to undertake actions. A proper documentation of different fire issues, and their evolution, will allow their integration into appropriate policies, whether specific to fire management, or complementary to other policies in areas such as forestry, rangeland, biodiversity, land tenure, etc. Strategies are found at all levels of fire management. They provide a shorter-term framework (e.g. one to five years) to prioritise fire management activities. They involve the development of a clear set of objectives and a clear set of activities to achieve these objectives. They may also include research and training inputs required, in order to build capacity and to answer specific questions needed to improve fire management. The chosen strategy will result from a trade-off between priority fire management objectives and the available capacity to act (e.g. institutional framework, budget, staff, etc.), and will lead towards a better allocation of resources for fire management operations to achieve specific objectives. One example in achieving an objective of conserving biotic diversity may be the implementation of a patch-mosaic burning system (Brockett et al., 200 1 ) instead of a prescribed block burning system, based on an assumption that the former should better promote biodiversity in the long-term than the latter (Parr & Brockett, 1999). This strategy requires the implementation of early season fires to reduce the size of later season fires. The knowledge of population movements, new settlements or a coming El Nino season, should help focus the resources usage, as these factors might influence the proportion as well as the locations of area burned. Another strategy may be to prioritise the grading of fire lines earlier than usual based on information on high biomass accumulation. However, whatever the strategies, they need to be based on reliable information

8. Remote Sensing Of Vegetation Fires And Its Contribution To A Fire Management Information System

In the last decade, research has proven that remote sensing can provide very useful support to fire managers. This chapter provides an overview of the types of information remote sensing can provide to the fire community. First, it considers fire management information needs in the context of a fire management information system. An introduction to remote sensing then precedes a description of fire information obtainable from remote sensing data (such as vegetation status, active fire detection and burned areas assessment). Finally, operational examples in five African countries illustrate the practical use of remotely sensed fire information. As indicated in previous chapters, fire management usually comprises activities designed to control the frequency, area, intensity or impact of fire. These activities are undertaken in different institutional, economic, social, environmental and geographical contexts, as well as at different scales, from local to national. The range of fire management activities also varies considerably according to the management issues at stake, as well as the available means and capacity to act. Whatever the level, effective fire management requires reliable information upon which to base appropriate decisions and actions. Information will be required at many different stages of this fire management system. To illustrate this, we consider a typical and generic description of a fire management loop , as provided in Figure 8.1. Fire management objectives result from fire related knowledge . For example, they may relate to sound ecological reasons for prescribed burning in a particular land management context, or to frequent, uncontrolled fires threatening valuable natural or human resources. Whatever the issues, appropriate objectives require scientific knowledge (such as fire impact on ecosystems components, such as soil and vegetation), as well as up-to date monitoring information (such as vegetation status, fire locations, land use, socioeconomic context, etc.). Policies, generally at a national and governmental level, provide the official or legal long term framework (e.g. five to ten years) to undertake actions. A proper documentation of different fire issues, and their evolution, will allow their integration into appropriate policies, whether specific to fire management, or complementary to other policies in areas such as forestry, rangeland, biodiversity, land tenure, etc. Strategies are found at all levels of fire management. They provide a shorter-term framework (e.g. one to five years) to prioritise fire management activities. They involve the development of a clear set of objectives and a clear set of activities to achieve these objectives. They may also include research and training inputs required, in order to build capacity and to answer specific questions needed to improve fire management. The chosen strategy will result from a trade-off between priority fire management objectives and the available capacity to act (e.g. institutional framework, budget, staff, etc.), and will lead towards a better allocation of resources for fire management operations to achieve specific objectives. One example in achieving an objective of conserving biotic diversity may be the implementation of a patch-mosaic burning system (Brockett et al., 200 1 ) instead of a prescribed block burning system, based on an assumption that the former should better promote biodiversity in the long-term than the latter (Parr & Brockett, 1999). This strategy requires the implementation of early season fires to reduce the size of later season fires. The knowledge of population movements, new settlements or a coming El Nino season, should help focus the resources usage, as these factors might influence the proportion as well as the locations of area burned. Another strategy may be to prioritise the grading of fire lines earlier than usual based on information on high biomass accumulation. However, whatever the strategies, they need to be based on reliable information

Cellular iron governs the host response to malaria

Malaria and iron deficiency are major global health problems with extensive epidemiological overlap. Iron deficiency-induced anaemia can protect the host from malaria by limiting parasite growth. On the other hand, iron deficiency can significantly disrupt immune cell function. However, the impact of host cell iron scarcity beyond anaemia remains elusive in malaria. To address this, we employed a transgenic mouse model carrying a mutation in the transferrin receptor (TfrcY20H/Y20H), which limits the ability of cells to internalise iron from plasma. At homeostasis TfrcY20H/Y20H mice appear healthy and are not anaemic. However, TfrcY20H/Y20H mice infected with Plasmodium chabaudi chabaudi AS showed significantly higher peak parasitaemia and body weight loss. We found that TfrcY20H/Y20H mice displayed a similar trajectory of malaria-induced anaemia as wild-type mice, and elevated circulating iron did not increase peak parasitaemia. Instead, P. chabaudi infected TfrcY20H/Y20H mice had an impaired innate and adaptive immune response, marked by decreased cell proliferation and cytokine production. Moreover, we demonstrated that these immune cell impairments were cell-intrinsic, as ex vivo iron supplementation fully recovered CD4+ T cell and B cell function. Despite the inhibited immune response and increased parasitaemia, TfrcY20H/Y20H mice displayed mitigated liver damage, characterised by decreased parasite sequestration in the liver and an attenuated hepatic immune response. Together, these results show that host cell iron scarcity inhibits the immune response but prevents excessive hepatic tissue damage during malaria infection. These divergent effects shed light on the role of iron in the complex balance between protection and pathology in malaria

Evaluation of perturbed iron-homeostasis in a prospective cohort of patients with COVID-19

Background: Marked reductions in serum iron concentrations are commonly induced during the acute phase of infection. This phenomenon, termed hypoferremia of inflammation, leads to inflammatory anemia, but could also have broader pathophysiological implications. In patients with coronavirus disease 2019 (COVID-19), hypoferremia is associated with disease severity and poorer outcomes, although there are few reported cohorts. Methods: In this study, we leverage a well characterised prospective cohort of hospitalised COVID-19 patients and perform a set of analyses focussing on iron and related biomarkers and both acute severity of COVID-19 and longer-term symptomatology. Results: We observed no associations between acute serum iron and long-term outcomes (including fatigue, breathlessness or quality of life); however, lower haemoglobin was associated with poorer quality of life. We also quantified iron homeostasis associated parameters, demonstrating that among 50 circulating mediators of inflammation IL-6 concentrations were strongly associated with serum iron, consistent with its central role in inflammatory control of iron homeostasis. Surprisingly, we observed no association between serum hepcidin and serum iron concentrations. We also observed elevated erythroferrone concentrations in COVID-19 patients with anaemia of inflammation. Conclusions: These results enhance our understanding of the regulation and pathophysiological consequences of disturbed iron homeostasis during SARS-CoV-2 infection. </p

An Intraocular Pressure Polygenic Risk Score Stratifies Multiple Primary Open-Angle Glaucoma Parameters Including Treatment Intensity

Purpose: To examine the combined effects of common genetic variants associated with intraocular pressure (IOP) on primary open-angle glaucoma (POAG) phenotype using a polygenic risk score (PRS) stratification. Design: Cross-sectional study. Participants: For the primary analysis, we examined the glaucoma phenotype of 2154 POAG patients enrolled in the Australian and New Zealand Registry of Advanced Glaucoma, including patients recruited from the United Kingdom. For replication, we examined an independent cohort of 624 early POAG patients. Methods Using IOP genome-wide association study summary statistics, we developed a PRS derived solely from IOP-associated variants and stratified POAG patients into 3 risk tiers. The lowest and highest quintiles of the score were set as the low- and high-risk groups, respectively, and the other quintiles were set as the intermediate risk group. Main Outcome Measures: Clinical glaucoma phenotype including maximum recorded IOP, age at diagnosis, number of family members affected by glaucoma, cup-to-disc ratio, visual field mean deviation, and treatment intensity. Results: A dose–response relationship was found between the IOP PRS and the maximum recorded IOP, with the high genetic risk group having a higher maximum IOP by 1.7 mmHg (standard deviation [SD], 0.62 mmHg) than the low genetic risk group (P = 0.006). Compared with the low genetic risk group, the high genetic risk group had a younger age of diagnosis by 3.7 years (SD, 1.0 years; P < 0.001), more family members affected by 0.46 members (SD, 0.11 members; P < 0.001), and higher rates of incisional surgery (odds ratio, 1.5; 95% confidence interval, 1.1–2.0; P = 0.007). No statistically significant difference was found in mean deviation. We further replicated the maximum IOP, number of family members affected by glaucoma, and treatment intensity (number of medications) results in the early POAG cohort (P ≤ 0.01). Conclusions: The IOP PRS was correlated positively with maximum IOP, disease severity, need for surgery, and number of affected family members. Genes acting via IOP-mediated pathways, when considered in aggregate, have clinically important and reproducible implications for glaucoma patients and their close family members