Retinal regions shape human and murine Müller cell proteome profile and functionality.

The human macula is a highly specialized retinal region with pit-like morphology and rich in cones. How Müller cells, the principal glial cell type in the retina, are adapted to this environment is still poorly understood. We compared proteomic data from cone- and rod-rich retinae from human and mice and identified different expression profiles of cone- and rod-associated Müller cells that converged on pathways representing extracellular matrix and cell adhesion. In particular, epiplakin (EPPK1), which is thought to play a role in intermediate filament organization, was highly expressed in macular Müller cells. Furthermore, EPPK1 knockout in a human Müller cell-derived cell line led to a decrease in traction forces as well as to changes in cell size, shape, and filopodia characteristics. We here identified EPPK1 as a central molecular player in the region-specific architecture of the human retina, which likely enables specific functions under the immense mechanical loads in vivo.

A tree of life? Multivariate logistic outcome-prediction in disorders of consciousness.

Background: Clinical outcome of patients with disorders of consciousness (DOC) is seen as generally very poor. Here, we specify individual outcome chances for patients with DOC on the basis of clinical and event-related-potentials (ERPs) data and identify subgroups, who vary substantially regarding their outcome chances.Methods: We employed data from 102 patients and used standard clinical protocol data (age, etiology, diagnosis, gender), sensory (N100, Mismatch-Negativity) and cognitive (P300, N400) ERPs to predict patients' recovery rates.Results: Two significant prediction models emerged: In both, subgroups of patients with good (51%, tree 1) to very good recovery chances (97%, tree 2) could be identified. The first model was obtained from standard clinical data. The second model included cognitive ERPs and resulted in considerably better patient classification. Moreover, when taking cognitive ERPs into account, the standard protocol data did not add further significant information, neither did sensory ERPs.Conclusion: The presented information about outcome chances of individual patients with DOC will be vital for these patients and critical for clinical professionals who have to direct specialized treatments and council relatives. Legal guardians and families, in turn, need to know what to expect in the future in order to prepare for the challenges ahead.

Sensing hydrocarbons with interband cascade lasers and substrate-integrated hollow waveguides.

Tunable diode laser absorption spectroscopy (TDLAS) is an excellent analytical technique for gas sensing applications. In situ sensing of relevant hydrocarbon gases is of substantial interest for a variety of in-field scenarios including environmental monitoring and process analysis, ideally providing accurate, molecule specific, and rapid information with minimal sampling requirements. Substrate-integrated hollow waveguides (iHWGs) have demonstrated superior properties for gas sensing applications owing to minimal sample volumes required while simultaneously serving as efficient photon conduits. Interband cascade lasers (ICLs) are recently emerging as mid-infrared light sources operating at room temperature, with low power consumption, and providing excellent potential for integration. Thereby, portable and handheld mid-infrared sensing devices are facilitated. Methane (CH4) is among the most frequently occurring, and thus, highly relevant hydrocarbons requiring in situ emission monitoring by taking advantage of its distinct molecular absorption around 3 µm. Here, an efficient combination of iHWGs with ICLs is presented providing a methane sensor calibrated in the range of 100 to 2000 ppmv with a limit of detection at 38 ppmv at the current stage of development. Furthermore, a measurement precision of 0.62 ppbv during only 1 s of averaging time has been demonstrated, thereby rendering this sensor concept useful for in-line and on-site emission monitoring and process control applications.

Epiplakin attenuates experimental mouse liver injury by chaperoning keratin reorganization.

BACKGROUND & AIMS: Epiplakin is a member of the plakin protein family and exclusively expressed in epithelial tissues where it binds to keratins. Epiplakin-deficient (Eppk1(-/-)) mice displayed no obvious spontaneous phenotype, but their keratinocytes showed a faster keratin network breakdown in response to stress. The role of epiplakin in the stressed liver remained to be elucidated. METHODS: Wild-type (WT) and Eppk1(-/-) mice were subjected to common bile duct ligation (CBDL) or fed with a 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-containing diet. The importance of epiplakin during keratin reorganization was assessed in primary hepatocytes. RESULTS: Our experiments revealed that epiplakin is expressed in hepatocytes and cholangiocytes, and binds to keratin 8 (K8) and K18 via multiple domains. In several liver stress models epiplakin and K8 genes displayed identical expression patterns and transgenic K8 overexpression resulted in elevated hepatic epiplakin levels. After CBDL and DDC treatment, Eppk1(-/-) mice developed a more pronounced liver injury and their livers contained larger amounts of hepatocellular keratin granules, indicating impaired disease-induced keratin network reorganization. In line with these findings, primary Eppk1(-/-) hepatocytes showed increased formation of keratin aggregates after treatment with the phosphatase inhibitor okadaic acid, a phenotype which was rescued by the chemical chaperone trimethylamine N-oxide (TMAO). Finally, transfection experiments revealed that Eppk1(-/-) primary hepatocytes were less able to tolerate forced K8 overexpression and that TMAO treatment rescued this phenotype. CONCLUSION: Our data indicate that epiplakin plays a protective role during experimental liver injuries by chaperoning disease-induced keratin reorganization.

Targeted proteolysis of plectin isoform 1a accounts for hemidesmosome dysfunction in mice mimicking the dominant skin blistering disease EBS-Ogna.

Autosomal recessive mutations in the cytolinker protein plectin account for the multisystem disorders epidermolysis bullosa simplex (EBS) associated with muscular dystrophy (EBS-MD), pyloric atresia (EBS-PA), and congenital myasthenia (EBS-CMS). In contrast, a dominant missense mutation leads to the disease EBS-Ogna, manifesting exclusively as skin fragility. We have exploited this trait to study the molecular basis of hemidesmosome failure in EBS-Ogna and to reveal the contribution of plectin to hemidesmosome homeostasis. We generated EBS-Ogna knock-in mice mimicking the human phenotype and show that blistering reflects insufficient protein levels of the hemidesmosome-associated plectin isoform 1a. We found that plectin 1a, in contrast to plectin 1c, the major isoform expressed in epidermal keratinocytes, is proteolytically degraded, supporting the notion that degradation of hemidesmosome-anchored plectin is spatially controlled. Using recombinant proteins, we show that the mutation renders plectin's 190-nm-long coiled-coil rod domain more vulnerable to cleavage by calpains and other proteases activated in the epidermis but not in skeletal muscle. Accordingly, treatment of cultured EBS-Ogna keratinocytes as well as of EBS-Ogna mouse skin with calpain inhibitors resulted in increased plectin 1a protein expression levels. Moreover, we report that plectin's rod domain forms dimeric structures that can further associate laterally into remarkably stable (paracrystalline) polymers. We propose focal self-association of plectin molecules as a novel mechanism contributing to hemidesmosome homeostasis and stabilization.

Single mode quantum cascade lasers with shallow-etched distributed Bragg reflector.

We report the fabrication of single mode quantum cascade lasers using a shallow-etched distributed Bragg reflector as frequency selective element. Quasi-continuous single mode tuning over 15 cm-1 at room temperature and 25 cm-1 via temperature tuning at Peltier temperatures is demonstrated. The behavior of both electro-optic and spectral characteristics under variation of the segment currents is analyzed, showing a maximum peak output power at room temperature of 600 mW. Thermal crosstalk between the laser segments is investigated. The spectral resolution of a gas absorption experiment is determined to be better than 0.0078 cm-1.

Comparative genomics reveals evolutionary loss of epiplakin in cetaceans.

The adaptation of vertebrates to different environments was associated with changes in the molecular composition and regulation of epithelia. Whales and dolphins, together forming the clade cetaceans, have lost multiple epithelial keratins during or after their evolutionary transition from life on land to life in water. It is unknown whether the changes in keratins were accompanied by gain or loss of cytoskeletal adapter proteins of the plakin family. Here we investigated whether plakin proteins are conserved in cetaceans and other vertebrates. Comparative analysis of genome sequences showed conservation of dystonin, microtubule actin crosslinking factor 1 (MACF1), plectin, desmoplakin, periplakin and envoplakin in cetaceans. By contrast, EPPK1 (epiplakin) was disrupted by inactivating mutations in all cetaceans investigated. Orthologs of EPPK1 are present in bony and cartilaginous fishes and tetrapods, indicating an evolutionary origin of EPPK1 in a common ancestor of jawed vertebrates (Gnathostomes). In many vertebrates, EPPK1 is flanked by an as-yet uncharacterized gene that encodes protein domains homologous to the carboxy-terminal segment of MACF1. We conclude that epiplakin, unlike other plakins, was lost in cetaceans.

A Review on Modeling Cure Kinetics and Mechanisms of Photopolymerization.

Photopolymerizations, in which the initiation of a chemical-physical reaction occurs by the exposure of photosensitive monomers to a high-intensity light source, have become a well-accepted technology for manufacturing polymers. Providing significant advantages over thermal-initiated polymerizations, including fast and controllable reaction rates, as well as spatial and temporal control over the formation of material, this technology has found a large variety of industrial applications. The reaction mechanisms and kinetics are quite complex as the system moves quickly from a liquid monomer mixture to a solid polymer. Therefore, the study of curing kinetics is of utmost importance for industrial applications, providing both the understanding of the process development and the improvement of the quality of parts manufactured via photopolymerization. Consequently, this review aims at presenting the materials and curing chemistry of such ultrafast crosslinking polymerization reactions as well as the research efforts on theoretical models to reproduce cure kinetics and mechanisms for free-radical and cationic photopolymerizations including diffusion-controlled phenomena and oxygen inhibition reactions in free-radical systems.

A Ca2+-Mediated Switch of Epiplakin from a Diffuse to Keratin-Bound State Affects Keratin Dynamics.

Keratins exert important structural but also cytoprotective functions. They have to be adaptable to support cellular homeostasis. Epiplakin (EPPK1) has been shown to decorate keratin filaments in epithelial cells and to play a protective role under stress, but the mechanism is still unclear. Using live-cell imaging of epithelial cells expressing fluorescently tagged EPPK1 and keratin, we report here an unexpected dynamic behavior of EPPK1 upon stress. EPPK1 was diffusely distributed throughout the cytoplasm and not associated with keratin filaments in living cells under standard culture conditions. However, ER-, oxidative and UV-stress, as well as cell fixation, induced a rapid association of EPPK1 with keratin filaments. This re-localization of EPPK1 was reversible and dependent on the elevation of cytoplasmic Ca2+ levels. Moreover, keratin filament association of EPPK1 led to significantly reduced keratin dynamics. Thus, we propose that EPPK1 stabilizes the keratin network in stress conditions, which involve increased cytoplasmic Ca2+.

Targeted inactivation of a developmentally regulated neural plectin isoform (plectin 1c) in mice leads to reduced motor nerve conduction velocity.

Cytolinker proteins stabilize cells mechanically, regulate cytoskeleton dynamics, and provide scaffolds for signaling molecules. For plectin, the prototype of these proteins, an unusual diversity of isoforms has been reported, which show distinct expression patterns, subcellular localizations, and functions. Plectin has been shown to have important functions in skin and muscle, but little is known about its role in neural cells. To address this issue, we generated two knock-out mouse lines, one which was selectively lacking plectin 1c (P1c), the major isoform expressed in neural cells, and another in which plectin was conditionally deleted in neuronal precursor cells. Using isoform-specific antibodies, we found P1c to be expressed late in development and to associate with postsynaptic dendrites of central nervous system neurons, motorneurons of spinal cord, sciatic nerve axons, and Schwann cells. Motor nerve conduction velocity was found significantly reduced in sciatic nerve from P1c-deficient as well as from conditional knock-out mice. This defect was traceable to an increased number of motor nerve fibers with small cross-sectional areas; the thicknesses of axons and of myelin sheaths were unaffected. This is the first report demonstrating an important role of plectin in a major nerve function.

DFB lasers between 760 nm and 16 µm for sensing applications.

Recent years have shown the importance of tunable semiconductor lasers in optical sensing. We describe the status quo concerning DFB laser diodes between 760 nm and 3,000 nm as well as new developments aiming for up to 80 nm tuning range in this spectral region. Furthermore we report on QCL between 3 µm and 16 µm and present new developments. An overview of the most interesting applications using such devices is given at the end of this paper.

Comparison and Impact of Different Fiber Debond Techniques on Fiber Reinforced Flexible Composites.

The focus of this paper is the realization and verification of a modified fiber bundle pull-out test setup to estimate the adhesion properties between threads and elastic matrix materials with a more realistic failure mode than single fiber debond techniques. This testing device including a modified specimen holder provides the basis for an adequate estimation of the interlaminar adhesion of fiber bundles including the opportunity of a faster, easier, and more economic handling compared to single fiber tests. The verification was done with the single-fiber and microbond test. Overall, the modified test setup showed the typical pull-out behavior, and the relative comparability between different test scales is given.

Viscoelastic Behavior of Glass-Fiber-Reinforced Silicone Composites Exposed to Cyclic Loading.

The aim of this work was to analyze the influence of fibers on the mechanical behavior of fiber-reinforced elastomers under cyclic loading. Thus, the focus was on the characterization of structure-property interactions, in particular the dynamic mechanical and viscoelastic behavior. Endless twill-woven glass fibers were chosen as the reinforcement, along with silicone as the matrix material. For the characterization of the flexible composites, a novel testing device was developed. Apart from the conventional dynamic mechanical analysis, in which the effect of the fiber orientation was also considered, modified step cycle tests were conducted under tensile loading. The material viscoelastic behavior was studied, evaluating both the stress relaxation response and the capability of the material to dissipate energy under straining. The effects of the displacement rate of the strain level, the amplitude of the strain applied in the loading-unloading step cycle test, and the number of the applied cycles were evaluated. The results revealed that an optimized fiber orientation leads to 30-fold enhanced stiffness, along with 10 times higher bearable stress. The findings demonstrated that tailored reinforced elastomers with endless fibers have a strong influence on the mechanical performance, affecting the structural properties significantly.

Tailored Interfaces in Fiber-Reinforced Elastomers: A Surface Treatment Study on Optimized Load Coupling via the Modified Fiber Bundle Debond Technique.

The interface between the reinforcement and surrounding matrix in a fibrous composite is decisive and critical for maintaining component performance, durability, and mechanical structure properties for load coupling assessment, especially for highly flexible composite materials. The clear trend towards tailored solutions reveals that an in-depth knowledge on surface treating methods to enhance the fiber-matrix interfacial interaction and adhesion properties for an optimized load transfer needs to be ensured. This research aims to quantify the effect of several surface treatments for glass fibers applied in endless fiber-reinforced elastomers with pronounced high deformations. Due to this, the glass fiber surface is directly modified with selected sizings, using a wet chemical treatment, and characterized according to chemical and mechanical aspects. For this purpose, the interfacial adhesion performance between fibers and the surrounding matrix material is investigated by a modified fiber pull-out device. The results clearly show that an optimized surface treatment improves the interface strength and chemical bonding significantly. The fiber pull-out test confirms that an optimized fiber-matrix interface can be enhanced up to 85% compared to standard surface modifications, which distinctly provides the basis of enhanced performances on the component level. These findings were validated by chemical analysis methods and corresponding optical damage analysis.

The Tension-Twist Coupling Mechanism in Flexible Composites: A Systematic Study Based on Tailored Laminate Structures Using a Novel Test Device.

The focus of this research is to quantify the effect of load-coupling mechanisms in anisotropic composites with distinct flexibility. In this context, the study aims to realize a novel testing device to investigate tension-twist coupling effects. This test setup includes a modified gripping system to handle composites with stiff fibers but hyperelastic elastomeric matrices. The verification was done with a special test plan considering a glass textile as reinforcing with different lay-ups to analyze the number of layers and the influence of various fiber orientations onto the load-coupled properties. The results demonstrated that the tension-twist coupling effect strongly depends on both the fiber orientation and the considered reinforcing structure. This enables twisting angles up to 25° with corresponding torque of about 82.3 Nmm, which is even achievable for small lay-ups with 30°/60° oriented composites with distinct asymmetric deformation. For lay-ups with ±45° oriented composites revealing a symmetric deformation lead, as expected, no tension-twist coupling effect was seen. Overall, these findings reveal that the described novel test device provides the basis for an adequate and reliable determination of the load-coupled material properties between stiff fibers and hyperelastic matrices.

Epiplakin is dispensable for skin barrier function and for integrity of keratin network cytoarchitecture in simple and stratified epithelia.

Epiplakin, a giant epithelial protein of >700 kDa, belongs to the plakin family of cytolinker proteins. It represents an atypical family member, however, as it consists entirely of plakin repeat domains but lacks any of the other domains commonly shared by plakins. Hence, its putative function as a cytolinker protein remains to be shown. To investigate epiplakin's biological role, we generated epiplakin-deficient mice by gene targeting in embryonic stem cells. Epiplakin-deficient mice were viable and fertile, without developing any discernible phenotype. Ultrastructurally, their epidermis revealed no differences compared to wild-type littermates, and cornified envelopes isolated from skin showed no alterations in shape or stability. Furthermore, neither embryonal formation nor later function of the epithelial barrier was affected. In primary cultures of epiplakin-deficient keratinocytes, the organization of actin filaments, microtubules, and keratin networks was found to be normal. Similarly, no alterations in keratin network organization were observed in simple epithelia of small intestine and liver or in primary hepatocytes. We conclude that, despite epiplakin's abundant and highly specific expression in stratified and simple epithelia, its absence in mice does not lead to severe skin dysfunctions, nor has it detectable consequences for keratin filament organization and cytoarchitecture of cells.

Heat Dissipation in Epoxy/Amine-Based Gradient Composites with Alumina Particles: A Critical Evaluation of Thermal Conductivity Measurements.

For the design of the next generation of microelectronic packages, thermal management is one of the key aspects and must be met by the development of polymers with enhanced thermal conductivity. While all polymer classes show a very low thermal conductivity, this shortcoming can be compensated for by the addition of fillers, yielding polymer-based composite materials with high thermal conductivity. The inorganic fillers, however, are often available only in submicron- and micron-scaled dimensions and, consequently, can sediment during the curing reaction of the polymer matrix. In this study, an epoxy/amine resin was filled with nano- and submicron-scaled alumina particles, yielding a gradient composite. It was found that the thermal conductivity according to laser flash analysis of a sliced specimen ranged from 0.25 to 0.45 W·m-1·K-1 at room temperature. If the thermal conductivity of an uncut specimen was measured with a guarded heat flow meter, the 'averaged' thermal conductivity was measured to be only 0.25 W·m-1·K-1. Finite element analysis revealed that the heat dissipation through a gradient composite was of intermediate speed in comparison with homogeneous composites exhibiting a non-gradient thermal conductivity of 0.25 and 0.45 W·m-1·K-1.

Compositionally Graded Absorber for Efficient and Stable Near-Infrared-Transparent Perovskite Solar Cells.

Compositional grading has been widely exploited in highly efficient Cu(In,Ga)Se2, CdTe, GaAs, quantum dot solar cells, and this strategy has the potential to improve the performance of emerging perovskite solar cells. However, realizing and maintaining compositionally graded perovskite absorber from solution processing is challenging. Moreover, the operational stability of graded perovskite solar cells under long-term heat/light soaking has not been demonstrated. In this study, a facile partial ion-exchange approach is reported to achieve compositionally graded perovskite absorber layers. Incorporating compositional grading improves charge collection and suppresses interface recombination, enabling to fabricate near-infrared-transparent perovskite solar cells with power conversion efficiency of 16.8% in substrate configuration, and demonstrate 22.7% tandem efficiency with 3.3% absolute gain when mechanically stacked on a Cu(In,Ga)Se2 bottom cell. Non-encapsulated graded perovskite device retains over 93% of its initial efficiency after 1000 h operation at maximum power point at 60 °C under equivalent 1 sun illumination. The results open an avenue in exploring partial ion-exchange to design graded perovskite solar cells with improved efficiency and stability.

Analyzing Dyadic Sequence Data-Research Questions and Implied Statistical Models.

The analysis of observational data is often seen as a key approach to understanding dynamics in romantic relationships but also in dyadic systems in general. Statistical models for the analysis of dyadic observational data are not commonly known or applied. In this contribution, selected approaches to dyadic sequence data will be presented with a focus on models that can be applied when sample sizes are of medium size (N = 100 couples or less). Each of the statistical models is motivated by an underlying potential research question, the most important model results are presented and linked to the research question. The following research questions and models are compared with respect to their applicability using a hands on approach: (I) Is there an association between a particular behavior by one and the reaction by the other partner? (Pearson Correlation); (II) Does the behavior of one member trigger an immediate reaction by the other? (aggregated logit models; multi-level approach; basic Markov model); (III) Is there an underlying dyadic process, which might account for the observed behavior? (hidden Markov model); and (IV) Are there latent groups of dyads, which might account for observing different reaction patterns? (mixture Markov; optimal matching). Finally, recommendations for researchers to choose among the different models, issues of data handling, and advises to apply the statistical models in empirical research properly are given (e.g., in a new r-package "DySeq").

Loss of organic cation transporter 3 (Oct3) leads to enhanced proliferation and hepatocarcinogenesis.

BACKGROUND: Organic cation transporters (OCT) are responsible for the uptake of a broad spectrum of endogenous and exogenous substrates. Downregulation of OCT is frequently observed in human hepatocellular carcinoma (HCC) and is associated with a poor outcome. The aim of our current study was to elucidate the impact of OCT3 on hepatocarcinogenesis. METHODS: Transcriptional and functional loss of OCT was investigated in primary murine hepatocytes, derived from Oct3-knockout (Oct3-/-; FVB.Slc22a3tm1Dpb ) and wildtype (WT) mice. Liver tumors were induced in Oct3-/- and WT mice with Diethylnitrosamine and Phenobarbital over 10 months and characterized macroscopically and microscopically. Key survival pathways were investigated by Western Blot analysis. RESULTS: Loss of Oct3-/- in primary hepatocytes resulted in significantly reduced OCT activity determined by [3H]MPP+ uptake in vivo. Furthermore, tumor size and quantity were markedly enhanced in Oct3-/- mice (p<0.0001). Oct3-/- tumors showed significant higher proliferation (p<0.0001). Ki-67 and Cyclin D expression were significantly increased in primary Oct3-/- hepatocytes after treatment with the OCT inhibitors quinine or verapamil (p<0.05). Functional inhibition of OCT by quinine resulted in an activation of c-Jun N-terminal kinase (Jnk), especially in Oct3-/- hepatocytes. CONCLUSION: Loss of Oct3 leads to enhanced proliferation and hepatocarcinogenesis in vivo.