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In October 2014, several specimens of Gobioninae were caught in the river Mur in Styria (Austria), later described as a new species of the genus Romanogobio. The species Romanogobio skywalkeri is to current knowledge endemic to the river Mur and of high conservation value. This study aims to investigate, for the first time, the microhabitat of R. skywalkeri in the upper river Mur. In winter 2016 and spring 2017 a sampling campaign was carried out to investigate the microhabitat of this species. In total, 60 adult and subadult individuals were caught. The microhabitat of these 60 individuals was assessed regarding water depth, flow velocity (v40 and v0) and substrate composition. R. skywalkeri at this point seems to be a highly rheophilic species with a narrow habitat preference. The preferred habitat found during our study consisted solely of riffle sections with fast flow velocity (0.45–1.3 m/s) and coarse substrate fractions, mostly meso‐ and macrolithal (6.3–40 cm). The current distribution range only covers about 100 km of the Mur between Fisching and Frohnleiten, which makes this endemic species highly vulnerable to habitat loss and alteration by anthropogenic impacts. [ABSTRACT FROM AUTHOR]

Concurrent in-situ four-point probe resistivity and high resolution synchrotron x-ray diffraction measurements were used to characterize room temperature recrystallization in electroplated Cu thin films. The x-ray data were used to obtain the variation with time of the integrated intensities and the peak-breadth from the Cu 111 and 200 reflections of the transforming grains. The variation of the integrated intensity and resistivity data with time was analyzed using the Johnson-Mehl-Avrami-Kolmogorov (JMAK) model. For both 111-textured and non-textured electroplated Cu films, four-point probe resistivity measurements yielded shorter transformation times than the values obtained from the integrated intensities of the corresponding Cu 111 reflections. In addition, the JMAK exponents fitted to the resistivity data were significantly smaller. These discrepancies could be explained by considering the different material volumes from which resistivity and diffraction signals originated, and the physical processes which linked these signals to the changes in the evolving microstructure. Based on these issues, calibration of the resistivity analysis with direct structural characterization techniques is recommended. [ABSTRACT FROM AUTHOR]

Time-resolved in situ x-ray diffraction measurements were used to study the room-temperature recrystallization kinetics of electroplated copper thin films with thicknesses between 400 and 1000 nm. The thinnest films exhibited limited recrystallization and subsequent growth of grains, while recrystallized grains in the thicker films grew until all as-plated microstructure was consumed. For all films, recrystallized grains that belonged to the majority texture component, <111>, started growing after the shortest incubation time. These grains exhibited volumetric growth until they achieved the film thickness. After this point the growth mode became planar, with the <111>-type grains growing in the plane of the film. Grains with the <100> direction normal to the film surface started growing after the <111>-type grains switched to planar growth. However, the planar growth of this texture component finished at the same time as the growth of the <111> grains. Profile fitting of the 111 peak permitted the separation of the diffraction signals from recrystallized and as-plated grain populations. The average strains in these two populations, calculated from the peak position of the corresponding {111} reflections, were different, indicating a heterogeneous stress state within this texture component. The increasing volume fraction of recrystallized <111> grains with time was monitored via the variation in the diffracted intensity. This variation could be represented by the Johnson-Mehl-Avrami-Kolmogorov model. [ABSTRACT FROM AUTHOR]

The current COVID-19 pandemia affects health care systems worldwide, however, to a variable extent depending on the caseload in each country. We aimed to provide a cross-sectional overview of current limitations or adaptions in lung transplant programs in Germany in from January to May 2020 due to the COVID-19 pandemia caused by severe acute respiratory syndrome coronavirus 2. A cross-sectional survey assessing various aspects of lung transplant activity was sent to all active lung transplant programs (n = 12) in Germany. Eight centers (66%) responded to the survey within the requested time frame. Four centers (50%) reported their activity is not restricted at all and four centers (50%) reported on moderate general limitations. The overall lung transplant activity in Germany from January to May 2020 contains 128 bilateral and 11 single lung transplantations, which is similar to the same period in the year 2019 (126 bilateral transplantations and 12 single lung transplantations). The results suggest that the influence of the COVID-19 pandemia on lung transplantation activity in Germany has been moderate so far. Nevertheless, adaptions such as extensive testing of donors and recipients were introduced to reduce the likelihood of infections and increase patient safety. Alertness to changes in COVID-19 reproduction rates might be required until effective antiviral therapy or vaccination is available. [ABSTRACT FROM AUTHOR]

Room temperature recrystallization responses of blanket electroplated Cu thin films deposited under various conditions were monitored in real-time using synchrotron X-ray diffraction. Nominal control of electroplating parameters such as plating current, bath chemistry, and plating time was found to be insufficient to ensure repeatability of the 50% recrystallization time, τ 50 , from sample to sample even though the thickness variations between samples were insignificant. Real-time X-ray analysis of samples from numerous electroplating baths showed that, for a given seed deposition process, a reliable estimation of τ 50 at room temperature can be obtained from the ratio of the integrated intensities of the 111 and 200 Cu reflections, I − 111 / I − 200 , of the electroplated film at time zero (immediately after plating). Among the plating parameters investigated seed-layer texture most influenced this ratio and, hence, the subsequent room temperature recrystallization behavior of the plated film. [ABSTRACT FROM AUTHOR]

Cardiac myosin binding protein C (cMyBP-C) is an essential regulator of cross bridge cycling. Through mechanisms that are incompletely understood the N-terminal domains (NTDs) of cMyBP-C can activate contraction even in the absence of calcium and can also inhibit cross bridge kinetics in the presence of calcium. In vitro studies indicated that the proline–alanine rich (p/a) region and C1 domain are involved in these processes, although effects were greater using human proteins compared to murine proteins (Shaffer et al. J Biomed Biotechnol 2010, 2010: 789798). We hypothesized that the p/a and C1 region are critical for the timing of contraction. In this study we tested this hypothesis using a mouse model lacking the p/a and C1 region (p/a-C1 −/− mice) to investigate the in vivo relevance of these regions on cardiac performance. Surprisingly, hearts of adult p/a-C1 −/− mice functioned normally both on a cellular and whole organ level. Force measurements in permeabilized cardiomyocytes from adult p/a-C1 −/− mice and wild type (Wt) littermate controls demonstrated similar rates of force redevelopment both at submaximal and maximal activation. Maximal and passive force and calcium sensitivity of force were comparable between groups as well. Echocardiograms showed normal isovolumetric contraction times, fractional shortening and ejection fraction, indicating proper systolic function in p/a-C1 −/− mouse hearts. p/a-C1 −/− mice showed a slight but significant reduction in isovolumetric relaxation time compared to Wt littermates, yet this difference disappeared in older mice (7–8 months of age). Moreover, stroke volume was preserved in p/a-C1 −/− mice, corroborating sufficient time for normal filling of the heart. Overall, the hearts of p/a-C1 −/− mice showed no signs of dysfunction even after chronic stress with an adrenergic agonist. Together, these results indicate that the p/a region and the C1 domain of cMyBP-C are not critical for normal cardiac contraction in mice and that these domains have little if any impact on cross bridge kinetics in mice. These results thus contrast with in vitro studies utilizing proteins encoding the human p/a region and C1 domain. More detailed insight in how individual domains of cMyBP-C function and interact, across species and over the wide spectrum of conditions in which the heart has to function, will be essential to a better understanding of how cMyBP-C tunes cardiac contraction. [ABSTRACT FROM AUTHOR]

The study outlines Martin Luther's theological concept of marriage as presented in his first extensive depiction concerning marriage, his Sermon „Vom ehelichen Leben" from 1522. According to Luther, the union of woman and man called marriage is based on divine creation and has its purpose in procreation to proclaim the gospel to the offspring. Thus marriage is the only place in where sexual intercourse is permitted in the order God in his grace gave to his human creatures. The union can be dissolved if certain circumstances hinder its purpose mentioned before. The guideline paper „Zwischen Autonomie und Angewiesenheit", published by the Evangelical Church in Germany (EKD) in 2013, also refers to Martin Luther and his concept of marriage intending to legitimate its own view about marriage, its place in contemporary society, and its raison d'être. By comparing Luthers concept with the paper of the EKD this study determines the relation between both voices. [ABSTRACT FROM AUTHOR]

Abstract: Meeting the demand for more food in the next 20–30years requires intensifying cereal cropping systems and increasing current yields to about 70–80% of the genetic yield potential. A dynamic and robust nutrient management approach such as site-specific nutrient management (SSNM) will be essential to increase yields and optimize profits while maintaining the productivity of these intensive cropping systems. SSNM has increased yield and profit in rice, maize, and wheat in major cropping systems in Asia; but, crop advisors have found it complex and difficult to implement in the field. Nutrient Expert (NE) was developed to provide crop advisors with a simpler and faster way to use SSNM. NE enables crop advisors to develop SSNM recommendations using existing site information. Nutrient Expert for Hybrid Maize (NEHM) increased yield and profit of farmers in Indonesia and the Philippines. In Indonesia, NEHM increased yield by 0.9tha−1, which increased profit by US$ 270ha−1 over farmer’s fertilizer practice (FFP). Compared with FFP, NEHM recommendations reduced fertilizer P (−4kgha−1), increased fertilizer K (+11kgha−1), and did not significantly change fertilizer N. In the Philippines, NEHM increased yield by 1.6tha−1 and profit by US$ 379ha−1 compared with FFP. Compared with FFP, NEHM gave higher rates of all three nutrients (+25kgNha−1, +4kgPha−1, and +11kgKha−1), which substantially increased fertilizer costs (US$ 64ha−1) but still increased profit by about six times the additional investment in fertilizer. NE accounts for the important factors affecting site-specific recommendations, which makes it a suitable starting point for developing nutrient management tools to reach more users. [Copyright &y& Elsevier]

The muscle-specific RING finger proteins MuRF1 and MuRF2 have been proposed to regulate protein degradation and gene expression in muscle tissues. We have tested the in vivo roles of MuRF1 and MuRF2 for muscle metabolism by using knockout (KO) mouse models. Single MuRF1 and MuRF2 KO mice are healthy and have normal muscles. Double knockout (dKO) mice obtained by the inactivation of all four MuRF1 and MuRF2 alleles developed extreme cardiac and milder skeletal muscle hypertrophy. Muscle hypertrophy in dKO mice was maintained throughout the murine life span and was associated with chronically activated muscle protein synthesis. During ageing (months 4–18), skeletal muscle mass remained stable, whereas body fat content did not increase in dKO mice as compared with wild-type controls. Other catabolic factors such as MAFbox/atrogin1 were expressed at normal levels and did not respond to or prevent muscle hypertrophy in dKO mice. Thus, combined inhibition of MuRF1/MuRF2 could provide a potent strategy to stimulate striated muscles anabolically and to protect muscles from sarcopenia during ageing. [ABSTRACT FROM AUTHOR]

The precise assembly of the highly organized filament systems found in muscle is critically important for its function. It has been hypothesized that nebulin, a giant filamentous protein extending along the entire length of the thin filament, provides a blueprint for muscle thin filament assembly. To test this hypothesis, we generated a KO mouse model to investigate nebulin functions in vivo. Nebulin KO mice assemble thin filaments of reduced lengths and ∼15% of their Z-disks are abnormally wide. Our data demonstrate that nebulin functions in vivo as a molecular ruler by specifying pointed- and barbed-end thin filament capping. Consistent with the shorter thin filament length of nebulin deficient mice, maximal active tension was significantly reduced in KO animals. Phenotypically, the murine model recapitulates human nemaline myopathy (NM), that is, the formation of nemaline rods combined with severe skeletal muscle weakness. The myopathic changes in the nebulin KO model include depressed contractility, loss of myopalladin from the Z-disk, and dysregulation of genes involved in calcium homeostasis and glycogen metabolism; features potentially relevant for understanding human NM. [ABSTRACT FROM AUTHOR]

Muscular dystrophy with myositis (mdm) is a recessive mouse mutation that is caused by a small deletion in the giant elastic muscle protein titin. Homozygous mdm/mdm mice develop a progressive muscular dystrophy, leading to death at ∼2 months of age. We surveyed the transcriptomes of skeletal muscles from 24 day old homozygous mdm/mdm and +/+ wild-type mice, an age when MDM animals have normal passive and active tensions and sarcomeric structure. Of the 12,488 genes surveyed (U74 affymetrix array), 75 genes were twofold to 30-fold differentially expressed, including CARP (cardiac ankyrin repeat protein), ankrd2/Arpp (a CARP-like protein) and MLP (muscle LIM protein), all of which associate with the titin filament system. The four genes most strongly affected (eightfold to 30-fold change) were all members of the CARP-regulated Nkx-2.5-dependent signal pathway, and CARP mRNA level was 30-fold elevated in MDM skeletal muscle tissues. The CARP protein overexpressed in MDM became associated with the I-band region of the sarcomere. The mdm mutation excises the C-terminal portion of titin''s N2A region, abolishing its interaction with p94/calpain-3 protease. Thus, the composition of the titin N2A protein complex is altered in MDM by incorporation of CARP and loss of p94/calpain-3. These changes were absent from the following control tissues (1) cardiac muscles from homozygous mdm/mdm animals, (2) skeletal and cardiac muscle from heterozygous mdm/+ animals, and (3) dystrophic muscles from MDX mice. Thus, the altered composition of the titin N2A complex is specific for the titin-based skeletal muscular dystrophy in MDM. [Copyright &y& Elsevier]

Recent advances in sensor technology for air pollution monitoring open new possibilities in the field of environmental epidemiology. The low spatial resolution of fixed outdoor measurement stations and modelling uncertainties currently limit the understanding of personal exposure. In this context, air quality sensor systems (AQSSs) offer significant potential to enhance personal exposure assessment. A pilot study was conducted to investigate the feasibility of the NO2 sensor model B43F and the particulate matter (PM) sensor model OPC-R1, both from Alphasense (UK), for use in epidemiological studies. Seven patients with chronic obstructive pulmonary disease (COPD) or asthma had built-for-purpose sensor systems placed inside and outside of their homes at fixed locations for one month. Participants documented their indoor activities, presence in the house, window status, and symptom severity and performed a peak expiratory flow test. The potential inhaled doses of PM2.5 and NO2 were calculated using different data sources such as outdoor data from air quality monitoring stations, indoor data from AQSSs, and generic inhalation rates (IR) or activity-specific IR. Moreover, the relation between indoor and outdoor air quality obtained with AQSSs, an indoor source apportionment study, and an evaluation of the suitability of the AQSS data for studying the relationship between air quality and health were investigated. The results highlight the value of the sensor data and the importance of monitoring indoor air quality and activity patterns to avoid exposure misclassification. The use of AQSSs at fixed locations shows promise for larger-scale and/or long-term epidemiological studies. [ABSTRACT FROM AUTHOR]

Airborne transmission by droplets and aerosols is important for the spread of viruses. Face masks are a well-established preventive measure, but their effectiveness for mitigating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission is still under debate. We show that variations in mask efficacy can be explained by different regimes of virus abundance and are related to population-average infection probability and reproduction number. For SARS-CoV-2, the viral load of infectious individuals can vary by orders of magnitude. We find that most environments and contacts are under conditions of low virus abundance (virus-limited), where surgical masks are effective at preventing virus spread. More-advanced masks and other protective equipment are required in potentially virus-rich indoor environments, including medical centers and hospitals. Masks are particularly effective in combination with other preventive measures like ventilation and distancing. [ABSTRACT FROM AUTHOR]

Implications for the academic and interpersonal development of children and adolescents underpin a global political consensus to maintain in‐classroom teaching during the ongoing COVID‐19 pandemic. In support of this aim, the WHO and UNICEF have called for schools around the globe to be made safer from the risk of COVID‐19 transmission. Detailed guidance is needed on how this goal can be successfully implemented in a wide variety of educational settings in order to effectively mitigate impacts on the health of students, staff, their families, and society. This review provides a comprehensive synthesis of current scientific evidence and emerging standards in relation to the use of layered prevention strategies (involving masks, distancing, and ventilation), setting out the basis for their implementation in the school environment. In the presence of increasingly infectious SARS‐Cov‐2 variants, in‐classroom teaching can only be safely maintained through a layered strategy combining multiple protective measures. The precise measures that are needed at any point in time depend upon a number of dynamic factors, including the specific threat‐level posed by the circulating variant, the level of community infection, and the political acceptability of the resultant risk. By consistently implementing appropriate prophylaxis measures, evidence shows that the risk of infection from in‐classroom teaching can be dramatically reduced. Current studies indicate that wearing high‐quality masks and regular testing are amongst the most important measures in preventing infection transmission; whilst effective natural and mechanical ventilation systems have been shown to reduce infection risks in classrooms by over 80%. [ABSTRACT FROM AUTHOR]

We introduce a novel time-resolved photoemission-based near-field illumination method, referred to as femtosecond normal-incidence photoemission microscopy (NI-PEEM). The change from the commonly used grazing-incidence to normal-incidence illumination geometry has a major impact on the achievable contrast and, hence, on the imaging potential of transient local near fields. By imaging surface plasmon polaritons in normal light incidence geometry, the observed fringe spacing directly resembles the wavelength of the plasmon wave. Our novel approach provides a direct descriptive visualization of SPP wave packets propagating across a metal surface. [ABSTRACT FROM AUTHOR]

Background: Ambient air pollution poses a major risk for the development and aggravation of respiratory diseases. Evidence suggests that even in low-level air pollution environments there is a risk for an increase in adverse respiratory symptoms. We examined whether variations in daily air pollution levels of nitrogen dioxide, ozone, or particulate matter in Berlin, Germany were associated with hospital admissions of chronic obstructive pulmonary disease (COPD) and asthma patients in a time series analysis.Methods: We calculated single and multi-pollutant models, investigated possible lags in effect, and analysed the influence of meteorological variables on the results. Data from January 2005 through December 2015 were used to quantify the concentration-response.Results: The risk ratio for asthma patients to be hospitalised on the same day of NO2 exposure was 1.101 per 10 µg/m3 NO2 increase (95% CI: 1.013 to 1.195), for COPD patients 1.123 (95% CI: 1.081 to 1.168). Neither the exposure to ozone (95% CI: 0.904 to 1.020), PM10 (95% CI: 0.990 to 1.127), nor PM2.5 (95% CI: 0.981 to 1.148) was associated with an increased risk ratio for asthma patients to be hospitalised. Risk ratios for the hospital admission of COPD patients were also not increased due to ozone (95% CI: 0.981 to 1.033), PM10 (95% CI: 0.988 to 1.032), or PM2.5 (95% CI: 0.966 to 1.019) exposure. The presented risk ratios and confidence intervals relate to the day of exposure. We found no increased hospitalisation risks with a delayed occurrence on subsequent days.Conclusions: A quantifiable, statistically significant increase in risk for asthma and COPD exacerbations owing to NO2 exposure at levels well below European regulatory limit values was observed. [ABSTRACT FROM AUTHOR]

Fertilizer K and P requirements for rice ( Oryza sativa L.) can be determined with site-specific nutrient management (SSNM) using estimated target yield, nutrient balances, and yield gains from added nutrient. We used the QUEFTS (QUantitative Evaluation of the Fertility of Tropical Soils) model with >8000 plot-level observations to estimate the relationship between grain yield and nutrient accumulation in above-ground dry matter of irrigated rice with harvest index ≥ 0.4. Predicted reciprocal internal efficiencies (RIEs) at 60–70% of yield potential corresponded to plant accumulation of 14.6 kg N, 2.7 kg P, and 15.9 kg K per tonne of grain yield. These RIEs enable determination of plant requirements for K and P and net output of K and P in harvested grain and removed crop residues at a target yield. Yield gains for nutrient applied to irrigated rice averaged 12% for K and 9% for P for 525 to 531 observations. For fields without certain yield gain, fertilizer K and P requirements can be determined by a partial maintenance approach (i.e., fertilizer input < output in nutrient balance), which considers nutrient supply mediated through soil processes and balances trade-offs between financial loss with full maintenance rates and risk of excessive nutrient depletion without nutrient application. When yield gains to an added nutrient are certain, partial maintenance plus yield gain can be used to determine fertilizer requirements. The SSNM-based approach and algorithms enable rapid development of field-specific K and P management. [ABSTRACT FROM AUTHOR]

MuRF1 is a member of the TRIM/RBCC superfamily, a gene family that encompasses a large variety of proteins, all sharing the conserved TRIM (Tripartite Motive) sequential array of RING, B-box, and coiled-coil domains. Within this family, MuRF1(also named TRIM63) is a specialized member that contributes to the development of muscle atrophy and sarcopenia. Here we studied MuRF1's role in muscle atrophy during muscle unloading induced by hindlimb suspension. Consistent with previous studies, we found that MuRF1 inactivation leads to an attenuated muscle atrophy response. The amount of protection was higher as compared to the denervation model, and within the 10 day-suspension period the soleus muscle was spared from atrophy in MuRF1-KO mice. Contractility studies on hindlimb suspended muscle tissues suggested that MuRF1's functions extend beyond muscle trophicity and implicate MuRF1 in muscle fatigue and MLC phosphorylation control: soleus muscle from MuRF1-KO mice fatigued significantly faster and in addition showed a reduced posttetanic twitch potentiation. Thus the present work further established the role of MuRF1 in muscle atrophy and for the first time shows that MuRF1 plays a role in muscle fatigue and twitch potentiation. [ABSTRACT FROM AUTHOR]

In several papers an increased quantity of cell-free plasma DNA as well as the presence of long DNA fragments in cell-free plasma and serum has been described. We isolated cell-free DNA from plasma, serum, and bronchial lavage supernatants from 33 lung cancer patients and 27 patients with a benign lung disease. The DNA was amplified by real-time PCR, and the quantity as well as the DNA integrity was determined. We did not find significant differences between the patient populations. Our results led us to conclude that this method is not useful in a diagnostic setting and is not able to differentiate between lung cancer patients and patients with a benign lung disease. [ABSTRACT FROM AUTHOR]

Abstract: During pathophysiological muscle wasting, a family of ubiquitin ligases, including muscle RING-finger protein-1 (MuRF1), has been proposed to trigger muscle protein degradation via ubiquitination. Here, we characterized skeletal muscles from wild-type (WT) and MuRF1 knockout (KO) mice under amino acid (AA) deprivation as a model for physiological protein degradation, where skeletal muscles altruistically waste themselves to provide AAs to other organs. When WT and MuRF1 KO mice were fed a diet lacking AA, MuRF1 KO mice were less susceptible to muscle wasting, for both myocardium and skeletal muscles. Under AA depletion, WT mice had reduced muscle protein synthesis, while MuRF1 KO mice maintained nonphysiologically elevated levels of skeletal muscle protein de novo synthesis. Consistent with a role of MuRF1 for muscle protein turnover during starvation, the concentrations of essential AAs, especially branched-chain AAs, in the blood plasma significantly decreased in MuRF1 KO mice under AA deprivation. To clarify the molecular roles of MuRF1 for muscle metabolism during wasting, we searched for MuRF1-associated proteins using pull-down assays and mass spectrometry. Muscle-type creatine kinase (M-CK), an essential enzyme for energy metabolism, was identified among the interacting proteins. Coexpression studies revealed that M-CK interacts with the central regions of MuRF1 including its B-box domain and that MuRF1 ubiquitinates M-CK, which triggers the degradation of M-CK via proteasomes. Consistent with MuRF1''s role of adjusting CK activities in skeletal muscles by regulating its turnover in vivo, we found that CK levels were significantly higher in the MuRF1 KO mice than in WT mice. Glucocorticoid modulatory element binding protein-1 and 3-hydroxyisobutyrate dehydrogenase, previously identified as potential MuRF1-interacting proteins, were also ubiquitinated MuRF1-dependently. Taken together, these data suggest that, in a multifaceted manner, MuRF1 participates in the regulation of AA metabolism, including the control of free AAs and their supply to other organs under catabolic conditions, and in the regulation of ATP synthesis under metabolic-stress conditions where MuRF1 expression is induced. [Copyright &y& Elsevier]

MURF-1, MURF-2 and MURF-3 are a specific class of RING finger proteins that are expressed in striated muscle tissues. MURF-1 has been suggested to act as an ubiquitin ligase, thereby controlling proteasome-dependent degradation of muscle proteins. Here, we performed yeast two-hybrid (YTH) screens of skeletal muscle cDNA libraries with MURF-1 baits to identify potential myocellular targets of MURF-1-dependent ubiquitination. This identified eight myofibrillar proteins as binding partners of MURF-1: titin, nebulin, the nebulin-related protein NRAP, troponin-I (TnI), troponin-T (TnT), myosin light chain 2 (MLC-2), myotilin and T-cap. YTH mating studies with MURF-1,2,3 baits indicated that these eight myofibrillar proteins are all targeted redundantly by both MURF-1 and MURF-2. Western blot studies on cardiac tissues from wild-type and MURF-1-deficient mice suggested that titin and nebulin were ubiquitinated at similar levels, and MLC-2 and TnI at reduced levels in MURF-1 KO mice. Mapping of the TnI and titin binding sites on MURF-1 peptide scans demonstrated their binding to motifs highly conserved between MURF-1 and MURF-2. Our data are consistent with a model in which MURF-1 and MURF-2 together target a specific set of myofibrillar proteins redundantly, most likely to control their ubiquitination-dependent degradation. Finally, our YTH screens identified the interaction of MURF-1 with 11 enzymes required for ATP/energy production in muscle including the mitochondrial ATP synthase and cytoplasmic creatine kinase. These data raise the possibility that MURF-1 may coordinately regulate the energy metabolism of mitochondrial and cytoplasmic compartments. [Copyright &y& Elsevier]

Titin (also known as connectin) is a giant protein with a wide range of cellular functions, including providing muscle cells with elasticity. Its physiological extension is largely derived from the PEVK segment, rich in proline (P), glutamate (E), valine (V), and lysine (K) residues. We studied recombinant PEVK molecules containing the two conserved elements: &ap;28-residue PEVK repeats and E-rich motifs. Single molecule experiments revealed that calcium-induced conformational changes reduce the bending rigidity of the PEVK fragments, and site-directed mutagenesis identified four glutamate residues in the E-rich motif that was studied (exon 129), as critical for this process. Experiments with muscle fibers showed that titin-based tension is calcium responsive. We propose that the PEVK segment contains E-rich motifs that render titin a calciumdependent molecular spring that adapts to the physiological state of the cell. [ABSTRACT FROM AUTHOR]

CARP, ankrd-2/Arpp, and DARP, are three members of a conserved gene family, referred to here as MARPs (muscle ankyrin repeat proteins). The expression of MARPs is induced upon injury and hypertrophy (CARP), stretch or denervation (ankrd2/Arpp), and during recovery following starvation (DARP), suggesting that they are involved in muscle stress response pathways. Here, we show that MARP family members contain within their ankyrin repeat region a binding site for the myofibrillar elastic protein titin. Within the myofibril, MARPs, myopalladin, and the calpain protease p94 appear to be components of a titin N2A-based signaling complex. Ultrastructural studies demonstrated that all three endogenous MARP proteins co-localize with I-band titin N2A epitopes in adult heart muscle tissues. In cultured fetal rat cardiac myocytes, passive stretch induced differential distribution patterns of CARP and DARP: staining for both proteins was increased in the nucleus and at the I-band region of myofibrils, while DARP staining also increased at intercalated discs. We speculate that the myofibrillar MARPs are regulated by stretch, and that this links titin-N2A-based myofibrillar stress/strain signals to a MARP-based regulation of muscle gene expression. [Copyright &y& Elsevier]

Nebulin is a giant (Mr 750–850 kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin (thin) filaments in vertebrate skeletal muscles. Nebulin''s potential role as a molecular template is based on its structural and biochemical properties. Its central ∼700 kDa portion associates with actin along the entire length of the thin filament, its N-terminal region extends to thin filament pointed ends, and ∼80 kDa of its C-terminal region integrates within the Z-line lattice. Here, we determined the exon/intron organization of the entire mouse nebulin gene, which contains 165 exons in a 202 kb segment. We identified 16 novel exons, 15 of which encode nebulin-repeat motifs (12 from its central region and 3 from its Z-line region). One novel exon shares high sequence homology to the 20 residue repeats of the tight-junction protein, ZO-1. RT-PCR analyses revealed that all 16 novel exons are expressed in mouse skeletal muscle. Surprisingly, we also amplified mRNA transcripts from mouse and human heart cDNA using primers designed along the entire length of nebulin. The expression of cardiac-specific nebulin transcripts was confirmed by in situ hybridization in fetal rat cardiomyocytes and in embryonic Xenopus laevis (frog) heart. On the protein level, antibodies specific for skeletal muscle nebulin''s N and C-terminal regions stained isolated rat cardiac myofibrils at the pointed and barbed ends of thin filaments, respectively. These data indicate a conserved molecular layout of the nebulin filament systems in both cardiac and skeletal myofibrils. We propose that thin filament length regulation in cardiac and skeletal muscles may share conserved nebulin-based mechanisms, and that nebulin isoform diversity may contribute to thin filament length differences in cardiac and skeletal muscle. [Copyright &y& Elsevier]

Aims The risk of mortality from the coronavirus disease that emerged in 2019 (COVID-19) is increased by comorbidity from cardiovascular and pulmonary diseases. Air pollution also causes excess mortality from these conditions. Analysis of the first severe acute respiratory syndrome coronavirus (SARS-CoV-1) outcomes in 2003, and preliminary investigations of those for SARS-CoV-2 since 2019, provide evidence that the incidence and severity are related to ambient air pollution. We estimated the fraction of COVID-19 mortality that is attributable to the long-term exposure to ambient fine particulate air pollution. Methods and results We characterized global exposure to fine particulates based on satellite data, and calculated the anthropogenic fraction with an atmospheric chemistry model. The degree to which air pollution influences COVID-19 mortality was derived from epidemiological data in the USA and China. We estimate that particulate air pollution contributed ∼15% (95% confidence interval 7–33%) to COVID-19 mortality worldwide, 27% (13 – 46%) in East Asia, 19% (8–41%) in Europe, and 17% (6–39%) in North America. Globally, ∼50–60% of the attributable, anthropogenic fraction is related to fossil fuel use, up to 70–80% in Europe, West Asia, and North America. Conclusion Our results suggest that air pollution is an important cofactor increasing the risk of mortality from COVID-19. This provides extra motivation for combining ambitious policies to reduce air pollution with measures to control the transmission of COVID-19. [ABSTRACT FROM AUTHOR]

Depth-dependent stress distributions within copper films possessing capping layers were measured during in situ thermal anneals. Glancing-incidence x-ray diffraction measurements of SiCxNyHz capped Cu films revealed that a strain gradient near the cap/Cu interface, created by constraint imposed by the cap during its deposition process, decreased as the sample temperature increased to 350 °C. Although the increase in sample temperature allowed Cu to approach its equilibrium lattice spacing at the cap deposition temperature and minimize the corresponding stress gradient, both the gradient and concomitant increase in residual bulk stress of the Cu film reappeared after cooling to room temperature. [ABSTRACT FROM AUTHOR]

The presence of capping layers deposited on copper films can create strain gradients through the Cu film thickness. Grazing-incidence and conventional x-ray diffraction measurements determined the in-plane stress of the Cu films as a function of depth. Cu films possessing a SiCxNyHz capping layer exhibited greater tensile stress near the cap than in the bulk, whereas Cu films possessing a CoWP film did not show a gradient. The constraint imposed by the SiCxNyHz cap during the cooling process from the cap deposition temperature (350 °C) leads to an increase in the in-plane stress of 180 MPa from the bulk value. [ABSTRACT FROM AUTHOR]

Aggregation of amyloid‐β (Aβ) that leads to the formation of plaques in Alzheimer's disease (AD) occurs through the stepwise formation of oligomers and fibrils. An earlier onset of aggregation is obtained upon intracerebral injection of Aβ‐containing brain homogenate into human APP transgenic mice that follows a prion‐like seeding mechanism. Immunoprecipitation of these brain extracts with anti‐Aβ oligomer antibodies or passive immunization of the recipient animals abrogated the observed seeding activity, although induced Aβ deposition was still evident. Here, we establish that, together with Aβ monomers, Aβ oligomers trigger the initial phase of Aβ seeding and that the depletion of oligomeric Aβ delays the aggregation process, leading to a transient reduction of seed‐induced Aβ deposits. This work extends the current knowledge about the role of Aβ oligomers beyond its cytotoxic nature by pointing to a role in the initiation of Aβ aggregation in vivo. We conclude that Aβ oligomers are important for the early initiation phase of the seeding process. [ABSTRACT FROM AUTHOR]

Time-resolved imaging of the propagation and interference of isolated ultrashort surface plasmon polariton wave packets is demonstrated using two photon photoemission microscopy. The group- and phase velocity of individual wave packets are determined experimentally. Using two counter-propagating surface plasmon polariton pulses, the transient formation of a standing surface plasmon polariton wave is imaged in time and space. We demonstrate that using a normal incidence geometry in time-resolved photoemission microscopy provides great advantages for in-situ imaging of surface plasmon polaritons in arbitrary plasmonic structures. A simple 1D wave-simulation is used to confirm the experimental results. [ABSTRACT FROM AUTHOR]

Reducing radiation damage is important and dietary antioxidants that can protect cells from such damage are of value. Dietary lycopene, a carotenoid found in tomatoes, protects human lymphoid cell membranes from damage by γ-radiation. We report that such protective effects are remarkably reduced as the oxygen concentration increases - near zero at 100% oxygen from fivefold protection at 20% oxygen and, dramatically, from 50-fold protection at 0% oxygen. Such huge differences imply that under higher oxygen concentrations lycopene could lead to improved cancer therapy using γ-radiation. The cells are not efficiently protected from the superoxide radical by lycopene. Noncellular studies suggest molecular mechanisms for the oxygen effect. [ABSTRACT FROM AUTHOR]

Cryogenic aerosol cleaning is a dry cleaning method used in the back end of line (BEOL) semiconductor manufacturing to remove defects from planar hydrophobic surfaces such as SiCOH and SiCxNyHz. Cryogenic aerosol cleaning is preferred over conventional wet cleaning methods as it is a non-contact cleaning method, which uses inert gases to generate sub-micrometer-sized solid aerosol particles that physically remove nanometer-sized contaminants on wafer surfaces. Particle removal mechanism involves detachment of the particles upon impact with aerosol, diffusion, and finally entrainment away from the wafer. In BEOL metal line patterning, particles on the dielectric isolation surfaces translate through the subsequent lithography and copper fill steps in to single or multiple metal line open defects that are yield killers. In this study, we show that the particle removal performance of the standard aerosol cleaning can be enhanced by pre-heating the wafer and use of a higher molecular weight inert gas, namely Ar, for aerosol generation. Both the addition of a Pre-heat step and the use of Ar as the aerosol source showed 47–52% reduction in single and multiple line opens detected through wafer electrical tests during high volume semiconductor manufacturing process. [ABSTRACT FROM AUTHOR]

Background: Bronchoscopic coil treatment has been shown to improve pulmonary function, exercise capacity, and quality of life in patients with severe emphysema. Objectives: To perform a meta-analysis of the results of four independent European clinical trials investigating this coil therapy for emphysema. Methods: Data on all patients included in the four European clinical trials were analyzed for efficacy and safety outcomes. Results: A total of 2,536 coils were placed during 259 procedures in 140 patients. A total of 37 chronic obstructive pulmonary disease exacerbations and 27 pneumonias were recorded as serious adverse events up to 1 year after treatment. The pneumothorax rate was 6.4%. Both 6 and 12 months after treatment, significant (all p < 0.001) improvements were observed for: forced expiratory volume in 1 s [+0.08 liters (±0.19) and +0.08 liters (±0.21)], residual volume [RV; -510 ml (±850) and -430 ml (±720)], 6-min walking distance [6MWD; +44.1 m (±69.8) and +38.1 m (±71.9)], and St. George's Respiratory Questionnaire score [SGRQ; -9.5 points (±14.3) and -7.7 points (±14.2)]. No differences in any outcome measures were observed between heterogeneous and homogeneous emphysema patients. Only a high baseline RV was found to be an independent predictor of successful treatment. Conclusions: Bronchoscopic coil treatment improves pulmonary function, 6MWD, and quality of life in patients with severe emphysema up to 1 year after treatment, independent of the distribution of the disease. © 2015 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Progression of idiopathic dilated cardiomyopathy (IDCM) is marked with extensive left ventricular remodeling whose clinical manifestations and molecular basis are poorly understood. We aimed to evaluate the clinical potential of titin ligands in monitoring progression of cardiac remodeling associated with end-stage IDCM. Expression patterns of 8 mechanoptotic machinery-associated titin ligands (ANKRD1, ANKRD2, TRIM63, TRIM55, NBR1, MLP, FHL2, and TCAP) were quantitated in endomyocardial biopsies from 25 patients with advanced IDCM. When comparing NYHA disease stages, elevated ANKRD1 expression levels marked transition from NYHA < IV to NYHA IV. ANKRD1 expression levels closely correlated with systolic strain depression and short E wave deceleration time, as determined by echocardiography. On molecular level, myocardial ANKRD1 and serum adiponectin correlated with low BAX/BCL-2 ratios, indicative of antiapoptotic tissue propensity observed during the worsening of heart failure. ANKRD1 is a potential marker for cardiac remodeling and disease progression in IDCM. ANKRD1 expression correlated with reduced cardiac contractility and compliance. The association of ANKRD1 with antiapoptotic response suggests its role as myocyte survival factor during late stage heart disease, warranting further studies on ANKRD1 during end-stage heart failure. [ABSTRACT FROM AUTHOR]

The intercalation of Eu underneath Gr on Ir(111) is comprehensively investigated by microscopic, magnetic, and spectroscopic measurements, as well as by density functional theory. Depending on the coverage, the intercalated Eu atoms form either a (2×2) or a (3√×3√)R30∘ superstructure with respect to Gr. We investigate the mechanisms of Eu penetration through a nominally closed Gr sheet and measure the electronic structures and magnetic properties of the two intercalation systems. Their electronic structures are rather similar. Compared to Gr on Ir(111), the Gr bands in both systems are essentially rigidly shifted to larger binding energies resulting in n doping. The hybridization of the Ir surface state S1 with Gr states is lifted, and the moiré superperiodic potential is strongly reduced. In contrast, the magnetic behavior of the two intercalation systems differs substantially, as found by x-ray magnetic circular dichroism. The (2×2) Eu structure displays plain paramagnetic behavior, whereas for the (3√×3√)R30∘ structure the large zero-field susceptibility indicates ferromagnetic coupling, despite the absence of hysteresis at 10 K. For the latter structure, a considerable easy-plane magnetic anisotropy is observed and interpreted as shape anisotropy. [ABSTRACT FROM AUTHOR]

Background The lung volume reduction (LVR) coil is a minimally invasive bronchoscopic nitinol device designed to reduce hyperinflation and improve elastic recoil in severe emphysema. We investigated the feasibility, safety and efficacy of LVR coil treatment in a prospective multicentre cohort trial in patients with severe emphysema. Methods Patients were treated in 11 centres. Safety was evaluated by recording all adverse events, efficacy by the St George's Respiratory Questionnaire (SGRQ) as primary endpoint, and pulmonary function testing, modified Medical Research Council dyspnoea score (mMRC) and 6-min walk distance (6MWD) up to 12 months after the final treatment. Results Sixty patients (60.9 ± 7.5 years, forced expiratory volume in 1 s (FEV1) 30.2 ± 6 . 3% pred) were bronchoscopically treated with coils (55 bilateral, 5 unilateral), with a median of 10 (range 5-15) coils per lobe. Within 30 days post-treatment, seven chronic obstructive pulmonary disease exacerbations (6.1%), six pneumonias (5.2%), four pneumothoraces (3.5%) and one haemoptysis (0.9%) occurred as serious adverse events. At 6 and 12 months, respectively, ASGRQ was --12.1±12.9 and -11.1±13.3 points, A6MWD was +29.7±74.1 m and +51.4±76 m, AFEV1 was +0.11 ±0.20 L and +0.11±0.30 L, and ARV (residual volume) was --0.65±0.90 L and --0.71±0.81 L (all p<0.01). Post hoc analyses showed significant responses for SGRQ, 6MWD and RV in patients with both heterogeneous and homogeneous emphysema. Conclusions LVR coil treatment results in significant clinical improvements in patients with severe emphysema, with a good safety profile and sustained results for up to 1 year. Trial registration number: NCT01328899. [ABSTRACT FROM AUTHOR]

Background: Summer heat waves with temperature extremes are becoming more frequent with growing numbers in morbidity and mortality in patients with respiratory diseases. The aim of this study was to evaluate the ramifications of heat stress (temperature >25 °C) on the health status of patients with pulmonary arterial hypertension (PAH). Methods: Fifteen patients with PAH (mean age = 66.7 ± 5.2 years) continuously wore an accelerometer from April 1 to September 30, 2011, and their daily step count was recorded. In addition, patients kept a diary to record data on seven standardized questions regarding their daily symptoms. Echocardiography, 6-minute walk test, NTproBNP, and Modified Medical Research Council Scale (MMRC) were assessed at baseline and at the end of the study after 6 months. Results: On heat-stress days, patients showed significantly more symptoms and lower total steps/day compared to thermal comfort days (3,995 ± 2,013 steps/day vs. 5,567 ± 2,434 steps/day, respectively; P < 0.001). There was a significant negative correlation between total steps/day and Temp ( R = −0.47; P < 0.001) and humidity ( R = −0.34; P < 0.001). A significant positive correlation was found between daily symptoms and Temp ( R = +0.79; P < 0.001) and humidity ( R = +0.23; P < 0.001). Conclusions: Heat stress is associated with a compromised clinical status in patients with PAH. Adaptation strategies must be implemented to prevent heart-related morbidity, including therapeutic adjustments and adequate room cooling in the patient's home and at the hospital. [ABSTRACT FROM AUTHOR]

Background: A home based tele-monitoring system was developed to assess the effects of heat stress (days > 25°C) on clinical and functional status in patients with chronic obstructive pulmonary disease (COPD).Methods: Sixty-two COPD patients (GOLD II-IV) were randomized into a tele-monitoring Group (TG, N = 32) or Control Group (CG, N = 30). Tele-monitoring included 1) daily clinical status (COPD Assessment Test-CAT), 2) daily lung function and 3) weekly 6-minute walk test (6MWT). Duration of monitoring lasted a total of nine months (9 M).Results: From June 1st-August 31st 2012, 32 days with heat stress (29.0 ± 2.5°C) were recorded and matched with 32 thermal comfort days (21.0 ± 2.9°C). During heat stress, the TG showed a significant reduction in lung function and exercise capacity (FEV1% predicted: 51.1 ± 7.2 vs. 57.7 ± 5.0%; P <0.001 and 6MWT performance: 452 ± 85 vs. 600 ± 76 steps; P <0.001) and increase in CAT scores (19.2 ± 7.9 vs. 16.2 ± 7.2; P <0.001).Over summer, significantly fewer TG patients suffered exacerbation of COPD compared to CG patients (3 vs. 14; P = 0.006). Over entire 9 M follow-up, the TG group had fewer exacerbations compared to CG (7 vs. 22; P = 0.012), shorter cumulative hospital stay (34 vs. 97 days) and 43% fewer specialist consultations (24. vs. 42; P = 0.04).Conclusion: Heat stress affects clinical and functional status in COPD. Tele-monitoring reduces exacerbation frequency and health care utilization during heat stress and other periods of the year.Trial Registration Drks-id: DRK00000705. [ABSTRACT FROM AUTHOR]

Background: Bronchoscopic thermal vapor ablation (BTVA) reduces lung volumes in emphysema patients by inducing a localized inflammatory response (LIR) leading to a healing process of fibrosis, but may also increase symptoms. Objectives: We sought to evaluate whether the clinical manifestation of LIR correlated with patient outcome. Methods: Respiratory adverse events and inflammatory markers were analyzed from a multicenter trial of BTVA in patients with upper-lobe-predominant emphysema. End points including changes in forced expiratory flow (FEV1), lobar volume, St. George's Respiratory Questionnaire (SGRQ), modified Medical Research Council (mMRC) and 6-minute-walk distance (6-MWD) were analyzed according to the presence or absence of a respiratory adverse event requiring treatment with an antibiotic or steroid. Results: Forty-four patients received BTVA. Increases of inflammatory markers were observed with a peak between the second and fourth week. Eighteen respiratory adverse events occurred in 16 patients within 30 days of BTVA, requiring antibiotics and/or steroids. These patients had significantly greater lobar volume reduction (65.3 vs. 33.4%, p = 0.007) and a change in residual volume at 12 months (-933 vs. 13 ml, p < 0.001) associated with a greater improvement of exercise capacity and health-related quality of life than patients without respiratory adverse events. Conclusion: Patients with more prominent respiratory symptoms in the first 30 days following BTVA experience greater efficacy. The clinical manifestations of the LIR are predictive of long-term clinical benefits. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: The prevalence of cardiovascular mortality is high in Chronic Obstructive Pulmonary Disease (COPD) and the identification of clinical parameters to improve risk stratification is of great interest. Objectives: This study aims to assess the predictive strength of daily walking activity on expression of cardiac biomarkers in patients with COPD. Methods: One hundred and five patients with COPD (66.1 ± 8.7 years of age) were prospectively analyzed. Daily walking activity was measured by means of accelerometry. Stepwise multivariate regression analyses were employed with either midregional proatrial natriuretic peptide (MRproANP) or plasma proadrenomedullin (MRproADM) as dependent variables, and age, age-adjusted Charlson score, Modified Medical Research Council Dyspnea Scale (MMRC), Saint Georges Respiratory Questionnaire total score and either total walk, steps per day or fast walk as covariates. Results: Independent predictors of MRproANP included age (p = 0.015) and either total walk or steps per day (both p < 0.0001). Total walk or steps per day were the only independent predictors of MRproADM (p < 0.0001). There was a significant negative correlation between fast walk and MMRC (R = -0.70; p < 0.001) and fast walk was only independently predictive of MRproANP but not MRproADM once MMRC was excluded from the list of covariates (p = 0.023 and p = 0.057, respectively). Conclusions: Daily walking activity independently predicts levels of circulating MRproANP and MRproADM in stable COPD patients, two prognostic biomarkers of cardiac distress associated with long-term survival upon exacerbation of COPD. Employing activity monitors in the stable state might simplify risk stratification in daily living. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Bronchoscopic thermal vapor ablation (BTVA) ablates emphysematous tissue through a localized inflammatory response followed by contractive fibrosis and tissue shrinkage leading to lung volume reduction that should not be influenced by collateral ventilation. Objectives: To determine the correlation of clinical data from a trial of BTVA with fissure integrity visually assessed by computed tomography (CT). Methods: We conducted a single-arm study of patients with upper lobe-predominant emphysema (n = 44). Patients received BTVA either to the right upper lobe or left upper lobe, excluding the lingula. Primary efficacy outcomes were forced expiratory volume in 1 s (FEV1) and St. George's Respiratory Questionnaire (SGRQ) at 6 months. Lobar volume reduction from CT was another efficacy outcome measurement. The fissure of the treated lobe was analyzed visually on preinterventional CT. Incompleteness of the small fissure, the upper half of the right large fissure and the whole left large fissure were estimated visually in 5% increments, and the relative amount of fissure incompleteness was calculated. Pearson correlation coefficients were calculated for the association between fissure incompleteness and change in efficacy outcomes (baseline to 6 months) of BTVA. Results: A total of 38 out of 44 patients (86%) had incompleteness in the relevant fissure. Calculated relevant fissure incompleteness was a mean of 13% of fissure integrity (range 0-63). Correlation coefficients for the association of incompleteness with outcomes were as follows: FEV1 = 0.17; lung volume reduction = -0.27; SGRQ score = -0.10; 6-min walk distance = 0.0; residual volume (RV) = -0.18, and RV/total lung capacity = -0.14. Conclusions: Lobar fissure integrity has no or minimal influence on BTVA-induced lung volume reduction and improvements in clinical outcomes. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Limb-girdle muscular dystrophy type 2A (LGMD2A) is a genetic disease that is caused by mutations in the calpain 3 gene (CAPN3), which encodes the skeletal muscle-specific calpain, calpain 3 (also known as p94). However, the precise mechanism by which p94 functions in the pathogenesis of this disease remains unclear. Here, using p94 knockin mice (termed herein p94KI mice) in which endogenous p94 was replaced with a proteolytically inactive but structurally intact p94:C129S mutant protein, we have demonstrated that stretch-dependent p94 distribution in sarcomeres plays a crucial role in the pathogenesis of LGMD2A. The p94KI mice developed a progressive muscular dystrophy, which was exacerbated by exercise. The exercise-induced muscle degeneration in p94KI mice was associated with an inefficient redistribution of p94:C129S in stretched sarcomeres. Furthermore, the p94KI mice showed impaired adaptation to physical stress, which was accompanied by compromised upregulation of muscle ankyrin-repeat protein-2 and hsp upon exercise. These findings indicate that the stretch-induced dynamic redistribution of p94 is dependent on its protease activity and essential to protect muscle from degeneration, particularly under conditions of physical stress. Furthermore, our data provide direct evidence that loss of p94 protease activity can result in LGMD2A and molecular insight into how this could occur. [ABSTRACT FROM AUTHOR]

Abstract: MuRF1 is a member of the RBCC (RING, B-box, coiled-coil) superfamily that has been proposed to act as an atrogin during muscle wasting. Here, we show that MuRF1 is preferentially induced in type-II muscle fibers after denervation. Fourteen days after denervation, MuRF1 protein was further elevated but remained preferentially expressed in type-II muscle fibers. Consistent with a fiber-type dependent function of MuRF1, the tibialis anterior muscle (rich in type-II muscle fibers) was considerably more protected in MuRF1-KO mice from muscle wasting when compared to soleus muscle with mixed fiber-types. We also determined fiber-type distributions in MuRF1/MuRF2 double-deficient KO (dKO) mice, because MuRF2 is a close homolog of MuRF1. MuRF1/MuRF2 dKO mice showed a profound loss of type-II fibers in soleus muscle. As a potential mechanism we identified the interaction of MuRF1/MuRF2 with myozenin-1, a calcineurin/NFAT regulator and a factor required for maintenance of type-II muscle fibers. MuRF1/MuRF2 dKO mice had lost myozenin-1 expression in tibialis anterior muscle, implicating MuRF1/MuRF2 as regulators of the calcineurin/NFAT pathway. In summary, our data suggest that expression of MuRF1 is required for remodeling of type-II fibers under pathophysiological stress states, whereas MuRF1 and MuRF2 together are required for maintenance of type-II fibers, possibly via the regulation of myozenin-1. [Copyright &y& Elsevier]

Aims: Familial hypertrophic cardiomyopathy (FHC) is frequently caused by cardiac myosin-binding protein C (cMyBP-C) gene mutations, which should result in C-terminal truncated mutants. However, truncated mutants were not detected in myocardial tissue of FHC patients and were rapidly degraded by the ubiquitin-proteasome system (UPS) after gene transfer in cardiac myocytes. Since the diversity and specificity of UPS regulation lie in E3 ubiquitin ligases, we investigated whether the muscle-specific E3 ligases atrogin-1 or muscle ring finger protein-1 (MuRF1) mediate degradation of truncated cMyBP-C. [ABSTRACT FROM PUBLISHER]

Background: This study aimed to show that SHOX2 DNA methylation is a tumor marker in patients with suspected lung cancer by using bronchial fluid aspirated during bronchoscopy. Such a biomarker would be clinically valuable, especially when, following the first bronchoscopy, a final diagnosis cannot be established by histology or cytology. A test with a low false positive rate can reduce the need for further invasive and costly procedures and ensure early treatment. Methods: Marker discovery was carried out by differential methylation hybridization (DMH) and real-time PCR. The real-time PCR based HeavyMethyl technology was used for quantitative analysis of DNA methylation of SHOX2 using bronchial aspirates from two clinical centres in a case-control study. Fresh-frozen and Saccomanno-fixed samples were used to show the tumor marker performance in different sample types of clinical relevance. Results: Valid measurements were obtained from a total of 523 patient samples (242 controls, 281 cases). DNA methylation of SHOX2 allowed to distinguish between malignant and benign lung disease, i.e. abscesses, infections, obstructive lung diseases, sarcoidosis, scleroderma, stenoses, at high specificity (68% sensitivity [95% CI 62-73%], 95% specificity [95% CI 91-97%]). Conclusions: Hypermethylation of SHOX2 in bronchial aspirates appears to be a clinically useful tumor marker for identifying subjects with lung carcinoma, especially if histological and cytological findings after bronchoscopy are ambiguous. [ABSTRACT FROM AUTHOR]

Nebulin is a giant filamentous F-actin-binding protein (∼800 kDa) that binds along the thin filament of the skeletal muscle sarcomere. Nebulin is one of the least well understood major muscle proteins. Although nebulin is usually viewed as a structural protein, here we investigated whether nebulin plays a role in muscle contraction by using skinned muscle fiber bundles from a nebulin knock-out (NEB KO) mouse model. We measured force-pCa (-log[Ca2+]) and force-ATPase relations, as well as the rate of tension re-development (ktr) in tibialis craniaus muscle fibers. To rule out any alterations in troponin (Tn) isoform expression and/or status of Tn phosphorylation, we studied fiber bundles that had been reconstituted with bacterially expressed fast skeletal muscle recombinant Tn. We also performed a detailed analysis of myosin heavy chain, myosin light chain, and myosin light chain 2 phosphorylation, which showed no significant differences between wild type and NEB KO. Our mechanical studies revealed that NEB 1(0 fibers had increased tension cost (5.9 versus 4.4 pmol millinewtons-1 mm-1s-1) and reductions in ktr (4.7 versus 7.3 s-1), calcium sensitivity (pCa50 5.74 versus 5.90), and cooperativity of activation (nH 3.64 versus 4.38). Our findings indicate the following: 1) in skeletal muscle nebulin increases thin filament activation, and 2) through altering cross-bridge cycling kinetics, nebulin increases force and efficiency of contraction. These novel properties of nebulin add a new level of understanding of skeletal muscle function and provide a mechanism for the severe muscle weakness in patients with nebulin-based nemaline myopathy. [ABSTRACT FROM AUTHOR]

Abstract: Background: Humoral circulating inflammatory cytokines such as tumor necrosis factor alpha (TNF-α) can impair skeletal muscle contractility. Furthermore, TNF-α expression correlates with elevated levels of atrogin-like muscle-specific ubiquitin E3 ligases, which are presumed to mediate muscle protein breakdown and atrophy. However, the casual relationships between MuRF1 and TNF-α and their relative contributions to muscle function impairment are not known. Methods: TNF-α or saline was injected into either C57Bl6 or MuRF1−/− mice. After 16–24 h, the expression of MuRF1 in skeletal muscle was quantified by quantitative reverse transcription–PCR and Western blot analysis. Muscle function was measured in an organ bath. To obtain a broader overview on potential alterations, two-dimensional gel electrophoresis was performed. Results: Wild-type animals injected with TNF-α had higher MuRF1 mRNA expression (saline versus TNF-α: 56.6±12.1 versus 133.6±30.3 arbitrary units; p <0.05) and protein expression (saline versus TNF-α: 0.38±0.11 versus 1.07±0.25 arbitrary units; p <0.05) as compared to saline-injected littermates. Furthermore, TNF-α reduced force development at 150 Hz by 25% in C57Bl6 animals (saline versus TNF-α: 2412±120 versus 1799±114 g/cm2; p <0.05), but not in MuRF1−/− mice (saline versus TNF-α: 2424±198 versus 2431±180 g/cm2; p =NS). Proteome analysis revealed a significant down-regulation of fast skeletal muscle troponin T in wild-type animals treated with TNF-α as compared to MuRF1−/− mice that received TNF-α. Conclusion: The results of this study demonstrate for the first time that TNF-α-induced reduction in skeletal muscle force development depends on the induction of the atrophy-related E3 ubiquitin ligase MuRF1. A link for the reduction in muscle force may be the TNF-α/MuRF1-mediated down-regulation of fast skeletal muscle troponin T. [Copyright &y& Elsevier]

Previous work suggested that altered Ca2+ homeostasis might contribute to dysfunction of nebulin-free muscle, as gene expression analysis revealed that the sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA)-inhibitor sarcolipin (SLN) is up-regulated >70-fold in nebulin knockout mice, and here we tested this proposal. We investigated SLN protein expression in nebulin-free and wild-type skeletal muscle, as well as expression of other Ca2+-handling proteins. Ca2+ uptake capacity was determined in isolated sarcoplasmic reticulum vesicles and in intact myofibers by measuring Ca2+ transients. Muscle contractile performance was determined in skinned muscle activated with exogenous Ca2+, as well as in electrically stimulated intact muscle. We found profound up-regulation of SLN protein in nebulin-free skeletal muscle, whereas expression of other Ca2+-handling proteins was not (calsequestrin and phospholamban) or was minimally (SERCA) affected. Speed of Ca2+ uptake was >3-fold decreased in sarcoplasmic reticulum vesicles isolated from nebulin-free muscle as well as in nebulin-free intact myofibers. Ca2+-activated stress in skinned muscle and stress produced by intact nebulin-free muscle were reduced to a similar extent compared with wild type. Half-relaxation time was significantly longer in nebulin-free compared with wild-type muscle. Thus, the present study demonstrates for the first time that nebulin might also be involved in physiological Ca2+ handling of the SR-myofibrillar system. [ABSTRACT FROM AUTHOR]