Gb3 Glycosphingolipids with Fluorescent Oligoene Fatty Acids: Synthesis and Phase Behavior in Model Membranes

Glycosphingolipids are involved in a number of physiological and pathophysiological processes, and they serve as receptors for a variety of bacterial toxins and viruses. To investigate their function in lipid membranes, fluorescently labeled glycosphingolipids are highly desirable. Herein, a synthetic route to access Gb₃ glycosphingolipids with fluorescently labeled fatty acids, consisting of pentaene and hexaene moieties either at the terminus or in the middle of the acyl chain, has been developed. The fluorescent properties of the Gb₃ derivatives were investigated in small unilamellar vesicles composed of a raft-like mixture. Phase-separated giant unilamellar vesicles (GUVs) allowed the quantification of the apparent partitioning coefficients of the Gb₃ compounds by means of confocal fluorescence laser scanning microscopy. The determined partition coefficients demonstrate that the Gb₃ derivatives are preferentially localized in the liquid-disordered (l_d) phase. To analyze whether the compounds behave like their physiological counterparts, Cy3-labeled (Cy: cyanine) Shiga toxin B subunits (STxB) were specifically bound to Gb₃-doped GUVs. However, the protein was favorably localized in the l_d phase, in contrast to results reported for STxB bound to naturally occurring Gb₃, which is discussed in terms of the packing density of the lipids in the liquid-ordered (l_o) phase.     

Materials Surgery – Reactivity Differences of Organic Groups in Hybrids

Synergistic properties in hybrid materials can emerge if the inorganic matrix has an electronic influence on the organic constituents and vice versa. This paper describes the drastic effect of SiO₂ in periodically ordered mesoporous organosilica materials (PMOs) on ethylene groups. A sophisticated, in situ solid‐state NMR spectroscopy study showed that the ozonolysis of ethylene groups follows an entirely different mechanism than is normal for organic, molecular groups. Ultimately, this leads to the topotactic transformation of the PMO material. Only if silicon is not in the alpha position to the double bond does it became possible to establish a new method to functionalize PMOs materials: the targeted scission of the ethylene group and the creation of functionalized pockets inside the pore walls of the mesoporous solid.     

High-precision diode-laser-based temperature measurement for air refractive index compensation

We present a laser-based system to measure the refractive index of air over a long path length. In optical distance measurements, it is essential to know the refractive index of air with high accuracy. Commonly, the refractive index of air is calculated from the properties of the ambient air using either Ciddor or Edlén equations, where the dominant uncertainty component is in most cases the air temperature. The method developed in this work utilizes direct absorption spectroscopy of oxygen to measure the average temperature of air and of water vapor to measure relative humidity. The method allows measurement of temperature and humidity over the same beam path as in optical distance measurement, providing spatially well-matching data. Indoor and outdoor measurements demonstrate the effectiveness of the method. In particular, we demonstrate an effective compensation of the refractive index of air in an interferometric length measurement at a time-variant and spatially nonhomogeneous temperature over a long time period. Further, we were able to demonstrate 7 mK RMS noise over a 67 m path length using a 120 s sample time. To our knowledge, this is the best temperature precision reported for a spectroscopic temperature measurement.     

Life margin assessment with Physics of Failure Tools application to BGA packages

Designers of electronic equipment are faced with the contractual obligation of guaranteeing that their products will fulfil the specifications on life duration. Classically, empirical methods based on databases and handbooks are used and features such as Mean Time Between Failure (MTBF) are provided. This paper presents how to use Physics of Failure (PoF) tools and results in order to provide the customer with a reliability feature, called life margin, based on Cycles To Failure (CTF) figures. The method defines and computes a life margin for 99% of the population in the given environmental life conditions. This feature is used to evaluate the different technological possibilities and decide the best trade-off to satisfy the customer’s requirement on life duration. The method has been applied to a given board and a given application, where the component, which limits reliability, has been identified as being packaged in a Ball Grid Array (BGA) package.     

Cancer-Associated Fibroblasts Regulate Kinase Activity in Mesothelioma Cell Lines via Paracrine Signaling and Thereby Dictate Cell Faith and Behavior

Background: Malignant pleural mesothelioma (MPM) has an infaust prognosis due to resistance to systemic treatment with platin-analoga. MPM cells modulate the immune response to their benefit. They release proinflammatory cytokines, such as TGF-ß, awakening resting fibrocytes that switch their phenotype into activated fibroblasts. Signaling interactions between cancer cells and cancer-associated fibroblasts (CAFs) play an integral part in tumor progression. This study aimed to investigate the role CAFs play in MPM progression, analyzing the impact this complex, symbiotic interaction has on kinase-related cell signaling in vitro. Methods: We simulated paracrine signaling in vitro by treating MPM cell lines with conditioned medium (CM) from fibroblasts (FB) and vice versa. NCI-H2052, MSTO-211H, and NCI-H2452 cell lines representing the three mayor MPM subtypes, while embryonal myofibroblast cell lines, IMR-90 and MRC-5, provide a CAFs-like phenotype. Subsequently, differences in proliferation rates, migratory behavior, apoptosis, necrosis, and viability were used as covariates for data analysis. Kinase activity of treated samples and corresponding controls were then analyzed using the PamStation12 platform (PamGene); Results: Treatment with myofibroblast-derived CM revealed significant changes in phosphorylation patterns in MPM cell lines. The observed effect differs strongly between the analyzed MPM cell lines and depends on the origin of CM. Overall, a much stronger effect was observed using CM derived from IMR-90 than MRC-5. The phosphorylation changes mainly affected the MAPK signaling pathway.; Conclusions: The factors secreted by myofibroblasts in fibroblasts CM significantly influence the phosphorylation of kinases, mainly affecting the MAPK signaling cascade in tested MPM cell lines. Our in vitro results indicate promising therapeutic effects by the use of MEK or ERK inhibitors and might have synergistic effects in combination with cisplatin-based treatment, improving clinical outcomes for MPM patients.     

Exploring the Potential of Norbornene‐Modified Mannosamine Derivatives for Metabolic Glycoengineering

Metabolic glycoengineering (MGE) allows the introduction of unnaturally modified carbohydrates into cellular glycans and their visualization through bioorthogonal ligation. Alkenes, for example, have been used as reporters that can react through inverse‐electron‐demand Diels–Alder cycloaddition with tetrazines. Earlier, norbornenes were shown to be suitable dienophiles; however, they had not previously been applied for MGE. We synthesized two norbornene‐modified mannosamine derivatives that differ in the stereochemistry at the norbornene (exo/endo linkage). Kinetic investigations revealed that the exo derivative reacts more than twice as rapidly as the endo derivative. Through derivatization with 1,2‐diamino‐4,5‐methylenedioxybenzene (DMB) we confirmed that both derivatives are accepted by cells and incorporated after conversion to a sialic acid. In further MGE experiments the incorporated sugars were ligated to a fluorophore and visualized through confocal fluorescence microscopy and flow cytometry.     

Inequalities for the Number of Walks in Graphs

We investigate the growth of the number w _k of walks of length k in undirected graphs as well as related inequalities. In the first part, we deduce the inequality w _2a+c ?w _2(a+b)+c ≤w _2a ?w _2(a+b+c), which we call the Sandwich Theorem. It unifies and generalizes an inequality by Lagarias et al. and an inequality by Dress and Gutman. In the same way, we derive the inequality w _2a+c (v,v)?w _2(a+b)+c (v,v)≤w _2a (v,v)?w _2(a+b+c)(v,v) for the number w _k (v,v) of closed walks of length k starting at a given vertex v. We then use a theorem of Blakley and Dixon to show $w_{2\ell+p}^{k}\leq w_{2\ell+pk}\cdot w_{2\ell}^{k-1}$ , which unifies and generalizes an inequality by Erd?s and Simonovits and, again, the inequality by Dress and Gutman. Both results can be translated directly into the corresponding forms using the higher order densities, which extends former results. In the second part, we provide a new family of lower bounds for the largest eigenvalue λ ₁ of the adjacency matrix based on closed walks. We apply the Sandwich Theorem to show monotonicity in this and a related family of lower bounds of Nikiforov. This leads to generalized upper bounds for the energy of graphs. In the third part, we demonstrate that a further natural generalization of the Sandwich Theorem is not valid for general graphs. We show that the inequality w _a+b ?w _a+b+c ≤w _a ?w _a+2b+c does not hold even in very restricted cases like w ₁?w ₂≤w ₀?w ₃ (i.e., $\bar{d}\cdot w_{2}\leq w_{3}$ ) in the context of bipartite or cycle free graphs. In contrast, we show that surprisingly this inequality is always satisfied for trees and we show how to construct worst-case instances (regarding the difference of both sides of the inequality) for a given degree sequence. We also prove the inequality w ₁?w ₄≤w ₀?w ₅ (i.e., $\bar{d}\cdot w_{4}\leq w_{5}$ ) for trees and conclude with a corresponding conjecture for longer walks.