DMT1 Protects Macrophages from Salmonella Infection by Controlling Cellular Iron Turnover and Lipocalin 2 Expression

Macrophages are at the center of innate pathogen control and iron recycling. Divalent metal transporter 1 (DMT1) is essential for the uptake of non-transferrin-bound iron (NTBI) into macrophages and for the transfer of transferrin-bound iron from the endosome to the cytoplasm. As the control of cellular iron trafficking is central for the control of infection with siderophilic pathogens such as Salmonella Typhimurium, a Gram-negative bacterium residing within the phagosome of macrophages, we examined the potential role of DMT1 for infection control. Bone marrow derived macrophages lacking DMT1 (DMT1fl/fl^LysMCre(+)) present with reduced NTBI uptake and reduced levels of the iron storage protein ferritin, the iron exporter ferroportin and, surprisingly, of the iron uptake protein transferrin receptor. Further, DMT1-deficient macrophages have an impaired control of Salmonella Typhimurium infection, paralleled by reduced levels of the peptide lipocalin-2 (LCN2). LCN2 exerts anti-bacterial activity upon binding of microbial siderophores but also facilitates systemic and cellular hypoferremia. Remarkably, nifedipine, a pharmacological DMT1 activator, stimulates LCN2 expression in RAW264.7 macrophages, confirming its DMT1-dependent regulation. In addition, the absence of DMT1 increases the availability of iron for Salmonella upon infection and leads to increased bacterial proliferation and persistence within macrophages. Accordingly, mice harboring a macrophage-selective DMT1 disruption demonstrate reduced survival following Salmonella infection. This study highlights the importance of DMT1 in nutritional immunity and the significance of iron delivery for the control of infection with siderophilic bacteria.     

Pre-Omicron Vaccine Breakthrough Infection Induces Superior Cross-Neutralization against SARS-CoV-2 Omicron BA.1 Compared to Infection Alone

SARS-CoV-2 variants raise concern because of their high transmissibility and their ability to evade neutralizing antibodies elicited by prior infection or by vaccination. Here, we compared the neutralizing abilities of sera from 70 unvaccinated COVID-19 patients infected before the emergence of variants of concern (VOCs) and of 16 vaccine breakthrough infection (BTI) cases infected with Gamma or Delta against the ancestral B.1 strain, the Gamma, Delta and Omicron BA.1 VOCs using live virus. We further determined antibody levels against the Nucleocapsid (N) and full Spike proteins, the receptor-binding domain (RBD) and the N-terminal domain (NTD) of the Spike protein. Convalescent sera featured considerable variability in the neutralization of B.1 and in the cross-neutralization of different strains. Their neutralizing capacity moderately correlated with antibody levels against the Spike protein and the RBD. All but one convalescent serum failed to neutralize Omicron BA.1. Overall, convalescent sera from patients with moderate disease had higher antibody levels and displayed a higher neutralizing ability against all strains than patients with mild or severe forms of the disease. The sera from BTI cases fell into one of two categories: half the sera had a high neutralizing activity against the ancestral B.1 strain as well as against the infecting strain, while the other half had no or a very low neutralizing activity against all strains. Although antibody levels against the spike protein and the RBD were lower in BTI sera than in unvaccinated convalescent sera, most neutralizing sera also retained partial neutralizing activity against Omicron BA.1, suggestive of a better cross-neutralization and higher affinity of vaccine-elicited antibodies over virus-induced antibodies. Accordingly, the IC50: antibody level ratios were comparable for BTI and convalescent sera, but remained lower in the neutralizing convalescent sera from patients with moderate disease than in BTI sera. The neutralizing activity of BTI sera was strongly correlated with antibodies against the Spike protein and the RBD. Together, these findings highlight qualitative differences in antibody responses elicited by infection in vaccinated and unvaccinated individuals. They further indicate that breakthrough infection with a pre-Omicron variant boosts immunity and induces cross-neutralizing antibodies against different strains, including Omicron BA.1.     

Structural Analysis of Mitochondrial Dynamics—From Cardiomyocytes to Osteoblasts: A Critical Review

Mitochondria play a crucial role in cell physiology and pathophysiology. In this context, mitochondrial dynamics and, subsequently, mitochondrial ultrastructure have increasingly become hot topics in modern research, with a focus on mitochondrial fission and fusion. Thus, the dynamics of mitochondria in several diseases have been intensively investigated, especially with a view to developing new promising treatment options. However, the majority of recent studies are performed in highly energy-dependent tissues, such as cardiac, hepatic, and neuronal tissues. In contrast, publications on mitochondrial dynamics from the orthopedic or trauma fields are quite rare, even if there are common cellular mechanisms in cardiovascular and bone tissue, especially regarding bone infection. The present report summarizes the spectrum of mitochondrial alterations in the cardiovascular system and compares it to the state of knowledge in the musculoskeletal system. The present paper summarizes recent knowledge regarding mitochondrial dynamics and gives a short, but not exhaustive, overview of its regulation via fission and fusion. Furthermore, the article highlights hypoxia and its accompanying increased mitochondrial fission as a possible link between cardiac ischemia and inflammatory diseases of the bone, such as osteomyelitis. This opens new innovative perspectives not only for the understanding of cellular pathomechanisms in osteomyelitis but also for potential new treatment options.     

Cocaprins, β-Trefoil Fold Inhibitors of Cysteine and Aspartic Proteases from Coprinopsis cinerea

We introduce a new family of fungal protease inhibitors with β-trefoil fold from the mushroom Coprinopsis cinerea, named cocaprins, which inhibit both cysteine and aspartic proteases. Two cocaprin-encoding genes are differentially expressed in fungal tissues. One is highly transcribed in vegetative mycelium and the other in the stipes of mature fruiting bodies. Cocaprins are small proteins (15 kDa) with acidic isoelectric points that form dimers. The three-dimensional structure of cocaprin 1 showed similarity to fungal β-trefoil lectins. Cocaprins inhibit plant C1 family cysteine proteases with K_i in the micromolar range, but do not inhibit the C13 family protease legumain, which distinguishes them from mycocypins. Cocaprins also inhibit the aspartic protease pepsin with K_i in the low micromolar range. Mutagenesis revealed that the β2-β3 loop is involved in the inhibition of cysteine proteases and that the inhibitory reactive sites for aspartic and cysteine proteases are located at different positions on the protein. Their biological function is thought to be the regulation of endogenous proteolytic activities or in defense against fungal antagonists. Cocaprins are the first characterized aspartic protease inhibitors with β-trefoil fold from fungi, and demonstrate the incredible plasticity of loop functionalization in fungal proteins with β-trefoil fold.     

Reconfigurable handling system

The demand for more versatile assembly and handling systems to facilitate customized production is gaining in importance, especially with regard to the constantly-increasing cost pressure, to expansion of the range of product versions and the shortening of innovation cycles. As a cost-effective approach for frequently changing assembly tasks, a novel manipulation concept has been developed by combining given robot technologies. This new handling system has a modular and adaptable layout, which consists of several mobile arms to manipulate the object in six-dimensional Cartesian space. After grasping, when the arms are attached to the object, the mechanical architecture is similar to parallel manipulators or cooperating robots. As the mounting and gripping points of the arms can easily be changed, the manipulator can be reconfigured so as to match the user’s preferences and needs. In addition to the kinematic adaption the regarding task, the hardware and new functions can be reconfigured as well. Contact elements, measurement and assembly devices as well as testing modules can easily be in integrated in the concept. A modular automatic control concept combined with a self-optimizing planning tool helps the user to find the optimal configuration and realize it in an economic way.     

Synthesis of crystalline metal nitride and metal carbide nanostructures by sol–gel chemistry

Attention toward nanosized metal nitrides and carbides is rapidly increasing thanks to their chemical characteristics that make them as valid and sustainable alternatives to noble metals in catalysis and to air-sensitive metals or oxides for applications under harsh conditions. They are mostly used as bulk phase or micron sized powders, due to an intrinsic difficulty to synthesize them as nanoparticles in a systematic and scalable fashion. However, nanosized metal nitrides and carbides could exhibit improved performances, e.g. in catalysis due to a higher surface area, and can be shaped more easily than corresponding larger grains for further specific applications. Recently, sol–gel chemistry has closed this gap and now enables the simple, cheap, and sustainable production of metal nitride and carbide nanoparticles.In the present review we give an overview on recent sol–gel based pathways for the synthesis of metal nitride and carbide nanoparticles, believing that a better knowledge of the potentialities of these still hardly touched materials stimulates research interest and applications.     

Comparison of different failure criteria for fiber-reinforced plastics in terms of fracture curves for arbitrary stress combinations

For fiber-reinforced plastics exists a big number of different criteria for the failure prediction. The intention of this paper is to compare the TSAI-HIL-, the LaRC04- and PUCK’s criterion in terms of their fracture curves for a unidirectional glass-fiber reinforced composite layer. Therefore after the implementation of these three criteria, the two-dimensional fracture curves for all possible stress combinations, which can be derived from a general spatial stress tensor, are computed. In this way, the characteristics of the criteria, similarities and differences and possible weak points become obvious.     

Characteristics of origin-destination pair traffic in Funet

In this paper we analyze measurements gathered from a 2.5~Gbps link in the Finnish university network (Funet) in 2004. The traffic is broken down into origin-destination (OD) pair components based on source and destination IP address. We study the traffic characteristics of these components, and identify four typical representative OD pairs. For these pairs we investigate the validity of a moving IID Gaussian model. We find that the statistical properties of these OD pairs differ significantly from each other, with only some of them close to Gaussian. The OD pairs are also found to have some cross-correlation between each other, contradicting an often made assumption about OD pair independence. Furthermore, the existence of a mean-variance relation between the OD pairs is studied. We find that there is a relation between mean and variance, but for some periods of time it is rather weak.