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Two or more attitudinal states are incoherent if and only if, necessarily, under conditions of full transparency, you are disposed to give up one of them. This is roughly the account of incoherence that has recently been put forward by Alex Worsnip (2018). In this article it is argued that Worsnip’s account of incoherence is incorrect. Not only does it fail to be fully general (i.e., it does not allow us to assess the coherence of all combinations of attitudes), but it also implies that an agent is necessarily incoherent just in virtue of being in one attitudinal state.

Here were report the combination of biocompatible click chemistry of ω-azidosphinganine with fluorescence microscopy and mass spectrometry as a powerful tool to elaborate the sphingolipid metabolism. The azide probe was efficiently synthesized over 13 steps starting from L-serine in an overall yield of 20% and was used for live-cell fluorescence imaging of the endoplasmic reticulum in living cells by bioorthogonal click reaction with a DBCO-labeled fluorophore revealing that the incorporated analogue is mainly localized in the endoplasmic membrane like the endogenous species. A LC-MS(/MS)-based microsomal in vitro assay confirmed that ω-azidosphinganine mimics the natural species enabling the identification and analysis of metabolic breakdown products of sphinganine as a key starting intermediate in the complex sphingolipid biosynthetic pathways. Furthermore, the sphinganine-fluorophore conjugate after click reaction was enzymatically tolerated to form its dihydroceramide and ceramide metabolites. Thus, ω-azidosphinganine represents a useful biofunctional tool for metabolic investigations both by in vivo fluorescence imaging of the sphingolipid subcellular localization in the ER and by in vitro high-resolution mass spectrometry analysis. This should reveal novel insights of the molecular mechanisms sphingolipids and their processing enzymes have e.g. in infection.

Expansion microscopy (ExM) enables super-resolution imaging of proteins and nucleic acids on conventional microscopes. However, imaging of details of the organization of lipid bilayers by light microscopy remains challenging. We introduce an unnatural short-chain azide- and amino-modified sphingolipid ceramide, which upon incorporation into membranes can be labeled by click chemistry and linked into hydrogels, followed by 4× to 10× expansion. Confocal and structured illumination microscopy (SIM) enable imaging of sphingolipids and their interactions with proteins in the plasma membrane and membrane of intracellular organelles with a spatial resolution of 10–20 nm. As our functionalized sphingolipids accumulate efficiently in pathogens, we use sphingolipid ExM to investigate bacterial infections of human HeLa229 cells by Neisseria gonorrhoeae, Chlamydia trachomatis and Simkania negevensis with a resolution so far only provided by electron microscopy. In particular, sphingolipid ExM allows us to visualize the inner and outer membrane of intracellular bacteria and determine their distance to 27.6 ± 7.7 nm., Imaging of lipid bilayers using light microscopy is challenging. Here the authors label cells using a short chain click-compatible ceramide to visualize mammalian and bacterial membranes with expansion microscopy.

Via regioselective Ir-catalyzed C–H borylation and subsequent reactions (i.e., viaBr4-Per or (BF3K)4-Per intermediates), we have introduced strong π-donors and acceptors at the 2,5,8,11-positions of perylene leading to unusual properties. Thus, incorporation of four donor diphenylamine (DPA) or four acceptor Bmes2 (mes = 2,4,6-Me3C6H2) moieties yields novel compounds which can be reversibly oxidized or reduced four times, respectively, an unprecedented behavior for monomeric perylene derivatives. Spectroelectrochemical measurements show NIR absorptions up to 3000 nm for the mono-cation radical of (DPA)4-Per and a strong electronic coupling over the perylene bridge was observed indicative of fully delocalized Robin-Day Class III behavior. Both (DPA)4-Per and (Bmes2)4-Per derivatives possess unusually long intrinsic singlet lifetimes (τ0), e.g., 94 ns for the former one. The compounds are emissive in solution, thin films, and the solid state, with apparent Stokes shifts that are exceptionally large for perylene derivatives. Transient absorption measurements on (DPA)4-Per reveal an additional excited state, with a long lifetime of 500 μs, which sensitizes singlet oxygen effectively.

Here were report the combination of biocompatible click chemistry of ω-azidosphinganine with fluorescence microscopy and mass spectrometry as a powerful tool to elaborate the sphingolipid metabolism. The azide probe was efficiently synthesized over 13 steps starting from l-serine in an overall yield of 20% and was used for live-cell fluorescence imaging of the endoplasmic reticulum in living cells by bioorthogonal click reaction with a DBCO-labeled fluorophore revealing that the incorporated analogue is mainly localized in the endoplasmic membrane like the endogenous species. A LC-MS(/MS)-based microsomal in vitro assay confirmed that ω-azidosphinganine mimics the natural species enabling the identification and analysis of metabolic breakdown products of sphinganine as a key starting intermediate in the complex sphingolipid biosynthetic pathways. Furthermore, the sphinganine-fluorophore conjugate after click reaction was enzymatically tolerated to form its dihydroceramide and ceramide metabolites. Thus, ω-azidosphinganine represents a useful biofunctional tool for metabolic investigations both by in vivo fluorescence imaging of the sphingolipid subcellular localization in the ER and by in vitro high-resolution mass spectrometry analysis. This should reveal novel insights of the molecular mechanisms sphingolipids and their processing enzymes have e.g. in infection.

Due to the lack of primary amino groups, lipids cannot be fixed by formaldehyde, glutaraldehyde and other chemical fixatives nor expanded using expansion microscopy (ExM). We present a protocol that uses unnatural short-chain azide- and amino-functionalized ceramides, which upon incorporation into cellular and bacterial membranes can be labeled by click chemistry and linked into hydrogels, followed by 4x to 10x expansion.

The analysis of protein enrichment in the detergent-resistant membranes (DRMs) isolated from immune cells enables us to analyze a link between the membrane lipid dynamics and cell activation. Here, we describe the fractionation of detergent-resistant membranes and the correlative analysis of the enrichment of T cell receptor (TCR) and ω-azido-modified synthetic ceramide in those fractions upon TCR stimulation.

Obligate intracellular bacteria like Chlamydia trachomatis undergo a complex developmental cycle between infectious non-replicative (EBs) and non-infectious replicative (RBs) forms. EBs shortly after entering a host cell transform to RBs, a crucial process in infection, initiating chlamydial replication. As Chlamydia fail to replicate outside the host cell it is currently unknown how the transition from EBs to RBs is initiated. Here we show in a cell-free approach in axenic media that uptake of glutamine by the bacteria is critical to initiate EB-RB transition. These bacteria utilize glutamine to synthesize cell wall peptidoglycan which has recently been detected in the septa of replicating intracellular Chlamydia. The increased requirement for glutamine in infected cells is achieved by reprogramming the glutamine metabolism in a c-Myc-dependent manner. Glutamine was effectively taken up by the glutamine transporter SLC1A5 and metabolized via glutaminase. Interference with this metabolic reprogramming limited growth of Chlamydia. Intriguingly, Chlamydia failed to produce progeny in SLC1A5 knockout mice. Thus, we report on the central role of glutamine for the development of an obligate intracellular pathogenic bacterium and the reprogramming of host glutamine metabolism, which may provide a basis for innovative anti-infective strategies.

Obligate intracellular bacteria such as Chlamydia trachomatis undergo a complex developmental cycle between infectious, non-replicative elementary-body and non-infectious, replicative reticulate-body forms. Elementary bodies transform to reticulate bodies shortly after entering a host cell, a crucial process in infection, initiating chlamydial replication. As Chlamydia fail to replicate outside the host cell, it is unknown how the replicative part of the developmental cycle is initiated. Here we show, using a cell-free approach in axenic media, that the uptake of glutamine by the bacteria is crucial for peptidoglycan synthesis, which has a role in Chlamydia replication. The increased requirement for glutamine in infected cells is satisfied by reprogramming the glutamine metabolism in a c-Myc-dependent manner. Glutamine is effectively taken up by the glutamine transporter SLC1A5 and metabolized via glutaminase. Interference with this metabolic reprogramming limits the growth of Chlamydia. Intriguingly, Chlamydia failed to produce progeny in SLC1A5-knockout organoids and mice. Thus, we report on the central role of glutamine for the development of an obligate intracellular pathogenic bacterium and the reprogramming of host glutamine metabolism, which may provide a basis for innovative anti-infection strategies.

Hussain claims that ethical consumers are subject to democratic requirements of morality, whereas ordinary price/quality consumers are exempt from these requirements. In this paper, we demonstrate that Hussain’s position is incoherent, does not follow from the arguments he offers for it, and entails a number of counterintuitive consequences.

Sphingolipids and glycosphingolipids can regulate cell recognition and signalling. Ceramide and sphingosine-1-phosphate are major players in the sphingolipid pathways and are involved in the initiation and regulation of signalling, apoptosis, stress responses and infection. Specific chemically synthesised sphingolipid derivatives containing small functionalities like azide or alkyne can mimic the biological properties of natural lipid species, which turns them into useful tools for the investigation of the highly complex sphingolipid metabolism by rapid and selective ‘click chemistry’ using sensitive tags like fluorophores. Subsequent analysis by various fluorescence microscopy techniques or mass spectrometry allows the identification and quantification of the corresponding sphingolipid metabolites as well as the research of associated enzymes. Here we present an overview of recent advances in the synthesis of ceramide and sphingosine analogues for bioorthogonal click reactions to study biosynthetic pathways and localization of sphingolipids for the development of novel therapeutics against lipid-dependent diseases.

We show that by judicious choice of substituents at the 2- and 7-positions of pyrene, the frontier orbital order of pyrene can be modified, giving enhanced control over the nature and properties of the photoexcited states and the redox potentials. Specifically, we introduced a julolidine-like moiety and Bmes2 (mes=2,4,6-Me3 C6 H2 ) as very strong donor (D) and acceptor (A), respectively, giving 2,7-D-π-D- and unsymmetric 2,7-D-π-A-pyrene derivatives, in which the donor destabilizes the HOMO-1 and the acceptor stabilizes the LUMO+1 of the pyrene core. Consequently, for 2,7-substituted pyrene derivatives, unusual properties are obtained. For example, very large bathochromic shifts were observed for all of our compounds, and unprecedented green light emission occurs for the D/D system. In addition, very high radiative rate constants in solution and in the solid state were recorded for the D-π-D- and D-π-A-substituted compounds. All compounds show reversible one-electron oxidations, and Jul2 Pyr exhibits a second oxidation, with the largest potential splitting (ΔE=440 mV) thus far reported for 2,7-substituted pyrenes. Spectroelectrochemical measurements confirm an unexpectedly strong coupling between the 2,7-substituents in our pyrene derivatives.

The requirements of rationality are fundamental in practical and theoretical philosophy. Nonetheless, there exists no correct account of what constitutes rational requirements. This paper attempts to provide a correct constitutive account of ‘rationality requires’. I argue that rational requirements are grounded in ‘necessary explanations of subjective incoherence’, as I shall put it. Rationality requires of you to X if and only if your rational capacities, in conjunction with the fact that you not-X, explain necessarily why you have a non-maximal degree of subjective coherence.

This paper argues that practical reasoning is a mental process which leads a person from a set of existent mental states to an intention. In Section 1, I defend this view against two other proposals according to which practical reasoning either concludes in an action itself or in a normative belief. Section 2 discusses the correctness of practical reasoning and explains how the correctness of instrumental reasoning can be explained by the logical relations that hold between the contents of the mental states. In Section 3, I explore the correctness of normative practical reasoning. I conclude with the sceptical view that correct practical reasoning cannot require us to intend to do what we believe we ought to do.

This paper discusses whether rationality, morality or prudence impose process-requirements upon us. It has been argued that process-requirements fulfil two essential functions within a system of rational, moral or prudential requirements. These functions are considered to prove the existence of process-requirements. First, process-requirements are deemed necessary to ensure that rationality, morality or prudence can guide our deliberations and actions. Second, their existence is regarded as essential for the correctness of our ordinary explanations of why a person possesses a certain degree of morality, rationality or prudence. However, I argue that these two functions are unable to show the existence of process-requirements. Instead, I propose a different essential function for process-requirements: they are necessary for attributing the correct degree of rationality, morality or prudence to a subject who is not entirely rational, moral or prudent. This function, I argue, necessitates the existence of process-requirements.

Davidson famously contended that all actions are movements of the agent's body. It has been objected, however, that Davidson's view is incompatible with his own definition of primitive actions. This paper argues that this objection is based on an incorrect reading of Davidson's argument. I will show that by reading "movements", in "all actions are bodily movements", transitively, Davidson's definition of primitive actions ceases to conflict with his thesis that all actions are bodily movements.

One popular line of argument put forward in support of the principle that the right is prior to the good is to show that teleological theories, which put the good prior to the right, lead to implausible normative results. There are situations, it is argued, in which putting the good prior to the right entails that we ought to do things that cannot be right for us to do. Consequently, goodness cannot (always) explain an action’s rightness. This indicates that what is right must be determined independently of the good.In this paper, I argue that these purported counterexamples to teleology fail to establish that the right must be prior to the good. In fact, putting the right prior to the good can lead to sets of ought statements which potentially conflict with the principle that ‘ought’ implies ‘can’. I argue that no plausible ethical theory can determine what is right independently of a notion of value or goodness. Every plausible ethical theory needs a mapping from goodness to rightness, which imp...