Circadian control of histone turnover during cardiac development and growth.

During postnatal cardiac hypertrophy, cardiomyocytes undergo mitotic exit, relying on DNA replication-independent mechanisms of histone turnover to maintain chromatin organization and gene transcription. In other tissues, circadian oscillations in nucleosome occupancy influence clock-controlled gene expression, suggesting a role for the circadian clock in temporal control of histone turnover and coordinated cardiomyocyte gene expression. We sought to elucidate roles for the master circadian transcription factor, Bmal1, in histone turnover, chromatin organization, and myocyte-specific gene expression and cell growth in the neonatal period. Bmal1 knockdown in neonatal rat ventricular myocytes decreased myocyte size, total cellular protein synthesis, and transcription of the fetal hypertrophic gene Nppb after treatment with serum or the α-adrenergic agonist phenylephrine. Depletion of Bmal1 decreased the expression of clock-controlled genes Per2 and Tcap, as well as Sik1, a Bmal1 target upregulated in adult versus embryonic hearts. Bmal1 knockdown impaired Per2 and Sik1 promoter accessibility as measured by micrococcal nuclease-quantitative PCR and impaired histone turnover as measured by metabolic labeling of acid-soluble chromatin fractions. Sik1 knockdown in turn decreased myocyte size, while simultaneously inhibiting natriuretic peptide B transcription and activating Per2 transcription. Linking these changes to chromatin remodeling, depletion of the replication-independent histone variant H3.3a inhibited myocyte hypertrophy and prevented phenylephrine-induced changes in clock-controlled gene transcription. Bmal1 is required for neonatal myocyte growth, replication-independent histone turnover, and chromatin organization at the Sik1 promoter. Sik1 represents a novel clock-controlled gene that coordinates myocyte growth with hypertrophic and clock-controlled gene transcription. Replication-independent histone turnover is required for transcriptional remodeling of clock-controlled genes in cardiac myocytes in response to growth stimuli.

Circadian Control of Histone Turnover During Cardiac Development and Growth.

Rationale: During postnatal cardiac hypertrophy, cardiomyocytes undergo mitotic exit, relying on DNA replication-independent mechanisms of histone turnover to maintain chromatin organization and gene transcription. In other tissues, circadian oscillations in nucleosome occupancy influence clock-controlled gene expression, suggesting an unrecognized role for the circadian clock in temporal control of histone turnover and coordinate cardiomyocyte gene expression. Objective: To elucidate roles for the master circadian transcription factor, Bmal1, in histone turnover, chromatin organization, and myocyte-specific gene expression and cell growth in the neonatal period. Methods and Results: Bmal1 knockdown in neonatal rat ventricular myocytes (NRVM) decreased myocyte size, total cellular protein, and transcription of the fetal hypertrophic gene Nppb following treatment with increasing serum concentrations or the α-adrenergic agonist phenylephrine (PE). Bmal1 knockdown decreased expression of clock-controlled genes Per2 and Tcap, and salt-inducible kinase 1 (Sik1) which was identified via gene ontology analysis of Bmal1 targets upregulated in adult versus embryonic hearts. Epigenomic analyses revealed co-localized chromatin accessibility and Bmal1 localization in the Sik1 promoter. Bmal1 knockdown impaired Per2 and Sik1 promoter accessibility as measured by MNase-qPCR and impaired histone turnover indicated by metabolic labeling of acid-soluble chromatin fractions and immunoblots of total and chromatin-associated core histones. Sik1 knockdown basally increased myocyte size, while simultaneously impairing and driving Nppb and Per2 transcription, respectively. Conclusions: Bmal1 is required for neonatal myocyte growth, replication-independent histone turnover, and chromatin organization at the Sik1 promoter. Sik1 represents a novel clock-controlled gene that coordinates myocyte growth with hypertrophic and clock-controlled gene transcription.

MBNL1 drives dynamic transitions between fibroblasts and myofibroblasts in cardiac wound healing.

Dynamic fibroblast to myofibroblast state transitions underlie the heart's fibrotic response. Because transcriptome maturation by muscleblind-like 1 (MBNL1) promotes differentiated cell states, this study investigated whether tactical control of MBNL1 activity could alter myofibroblast activity and fibrotic outcomes. In healthy mice, cardiac fibroblast-specific overexpression of MBNL1 transitioned the fibroblast transcriptome to that of a myofibroblast and after injury promoted myocyte remodeling and scar maturation. Both fibroblast- and myofibroblast-specific loss of MBNL1 limited scar production and stabilization, which was ascribed to negligible myofibroblast activity. The combination of MBNL1 deletion and injury caused quiescent fibroblasts to expand and adopt features of cardiac mesenchymal stem cells, whereas transgenic MBNL1 expression blocked fibroblast proliferation and drove the population into a mature myofibroblast state. These data suggest MBNL1 is a post-transcriptional switch, controlling fibroblast state plasticity during cardiac wound healing.

A metrologically traceable protocol for the quantification of trace metals in different types of microplastic.

The presence of microplastic (MP) particles in aquatic environments raised concern about possible enrichment of organic and inorganic pollutants due to their specific surface and chemical properties. In particular the role of metals within this context is still poorly understood. Therefore, the aim of this work was to develop a fully validated acid digestion protocol for metal analysis in different polymers, which is a prerequisite to study such interactions. The proposed digestion protocol was validated using six different certified reference materials in the microplastic size range consisting of polyethylene, polypropylene, acrylonitrile butadiene styrene and polyvinyl chloride. As ICP-MS/MS enabled time-efficient, sensitive and robust analysis of 56 metals in one measurement, the method was suitable to provide mass fractions for a multitude of other elements beside the certified ones (As, Cd, Cr, Hg, Pb, Sb, Sn and Zn). Three different microwaves, different acid mixtures as well as different temperatures in combination with different hold times were tested for optimization purposes. With the exception of Cr in acrylonitrile butadiene styrene, recovery rates obtained using the optimized protocol for all six certified reference materials fell within a range from 95.9% ± 2.7% to 112% ± 7%. Subsequent optimization further enhanced both precision and recoveries ranging from 103% ± 5% to 107 ± 4% (U; k = 2 (n = 3)) for all certified metals (incl. Cr) in acrylonitrile butadiene styrene. The results clearly show the analytical challenges that come along with metal analysis in chemically resistant plastics. Addressing specific analysis tools for different sorption scenarios and processes as well as the underlying kinetics was beyond this study's scope. However, the future application of the two recommended thoroughly validated total acid digestion protocols as a first step in the direction of harmonization of metal analysis in/on MP will enhance the significance and comparability of the generated data. It will contribute to a better understanding of the role of MP as vector for trace metals in the environment.

Substituting HF by HBF4- an optimized digestion method for multi-elemental sediment analysis via ICP-MS/MS.

Determination of elemental mass fractions in sediments plays a major role in evaluating the environmental status of aquatic ecosystems. Herewith, the optimization of a new total digestion protocol and the subsequent analysis of 48 elements in different sediment reference materials (NIST SRM 2702, GBW 07313, GBW 07311 and JMC-2) based on ICP-MS/MS detection is presented. The developed method applies microwave acid digestion and utilizes HBF4 as fluoride source for silicate decomposition. Similar to established protocols based on HF, HBF4 ensures the dissolution of the silicate matrix, as well as other refractory oxides. As HBF4 is not acutely toxic; no special precautions have to be made and digests can be directly measured via ICP-MS without specific sample inlet systems, evaporation steps or the addition of e.g. H3BO3, in order to mask excess HF. Different acid mixtures with and without HBF4 were evaluated in terms of digestion efficiency based on the trace metal recovery. The optimized protocol (5 mL HNO3, 2 mL HCL, 1 mL HBF4) allows a complete dissolution of the analyzed reference materials, as well as quantitative recoveries for a wide variety of certified analytes. Low recoveries for e.g. Sr, Ba and rare earth elements due to fluoride precipitation of HF-based digestions protocols, can be avoided by the usage of HBF4 instead. Based on the usage of high purity HBF4 all relevant trace, as well as matrix elements can be analyzed with sufficiently low LOQs (0.002 µg L-1 for U up to 6.7 µg L-1 for Al). In total, 34 elements were within a recovery range of 80%-120% for all three analyzed reference materials GBW 07313, GBW 07311 and JMC-2. 14 elements were outside a recovery range of 80%-120% for at least one of the analyzed reference materials.

Characterization of alloying components in galvanic anodes as potential environmental tracers for heavy metal emissions from offshore wind structures.

The impact of offshore constructions on the marine environment is unknown in many aspects. The application of Al- and Zn-based galvanic anodes as corrosion protection results in the continuous emission of inorganic matter (e.g. >80 kg Al-anode material per monopile foundation and year) into the marine environment. To identify tracers for emissions from offshore wind structures, anode materials (Al-based and Zn-based) were characterized for their elemental and isotopic composition. An acid digestion and analysis method for Al and Zn alloys was adapted and validated using the alloy CRMs ERM®-EB317 (AlZn6CuMgZr) and ERM®-EB602 (ZnAl4Cu1). Digests were measured for their elemental composition by ICP-MS/MS and for their Pb isotope ratios by MC ICP-MS. Ga and In were identified as potential tracers. Moreover, a combined tracer approach of the elements Al, Zn, Ga, Cd, In and Pb together with Pb isotope ratios is suggested for a reliable identification of offshore-wind-farm-induced emissions. In the Al anodes, the mass fractions were found to be >94.4% of Al, >26200 mg kg-1 of Zn, >78.5 mg kg-1 of Ga, >0.255 mg kg-1 of Cd, >143 mg kg-1 of In and >6.7 mg kg-1 of Pb. The Zn anodes showed mass fractions of >2160 mg kg-1 of Al, >94.5% of Zn, >1.31 mg kg-1 of Ga, >254 mg kg-1 of Cd, >0.019 mg kg-1 of In and >14.1 mg kg-1 of Pb. The n(208Pb)/n(206Pb) isotope ratios in Al anodes range from 2.0619 to 2.0723, whereas Zn anodes feature n(208Pb)/n(206Pb) isotope ratios ranging from 2.0927 to 2.1263.

Extreme spatial variation of Sr, Nd and Pb isotopic signatures and 48 element mass fractions in surface sediment of the Elbe River Estuary - Suitable tracers for processes in dynamic environments?

The Elbe River has been long considered as one of the most anthropogenically impacted rivers in Europe. Its estuary is characterized by strong tidal effects, continuous dredging and dumping of sediment, and intense ship traffic between the North Sea and the Port of Hamburg. The aim of this study was to elucidate if a combined multi-element fingerprinting and isotopic tracer approach represented a suitable tool to investigate transport and mixing processes of inorganic contaminants within a complex and highly dynamic estuarine environment. A total of 37 surface sediment samples from the tidal Elbe were characterized in a comprehensive survey by determining the mass fractions of 48 elements and the isotopic signatures of stable Sr, Nd and Pb. Statistical data analysis resolved four discrete clusters of sampling locations in the estuary: One cluster upstream of the city of Hamburg, two clusters within the mixing zone between Hamburg and the mouth of the Elbe Estuary and one cluster in the mouth of the Estuary. River sediment entering the estuary carry significantly higher loads of metals (e.g. Cu, Zn, Sb, Cd and Pb), which are rapidly "diluted" by lower elemental mass fractions in marine sediment on a remarkably small regional scale. The cluster within the mouth of the estuary is mainly characterized by extreme isotopic variations of n(208Pb)/n(204Pb) ranging from 38.67 ±â€¯0.15 to 73.86 ±â€¯0.29, beside high mass fractions of U, Th, and some rare-earth elements. Determined Pb isotope ratios are among the highest reported values for terrestrial materials. This study indicates the general potential of combined element fingerprinting and isotope tracer approaches to elucidate processes in complex river systems. Furthermore, it represents an initial characterization of the catchment area of the Elbe River as basis for future studies on river and harbor management.

How Far Does a Receptor Influence Vibrational Properties of an Odorant?

The biophysical mechanism of the sense of smell, or olfaction, is still highly debated. The mainstream explanation argues for a shape-based recognition of odorant molecules by olfactory receptors, while recent investigations suggest the primary olfactory event to be triggered by a vibrationally-assisted electron transfer reaction. We consider this controversy by studying the influence of a receptor on the vibrational properties of an odorant in atomistic details as the coupling between electronic degrees of freedom of the receptor and the vibrations of the odorant is the key parameter of the vibrationally-assisted electron transfer. Through molecular dynamics simulations we elucidate the binding specificity of a receptor towards acetophenone odorant. The vibrational properties of acetophenone inside the receptor are then studied by the polarizable embedding density functional theory approach, allowing to quantify protein-odorant interactions. Finally, we judge whether the effects of the protein provide any indications towards the existing theories of olfaction.

Providing multilingual access to health-related content.

Finding health-related content is not an easy task. People have to know what to search for, which medical terms to use, and where to find accurate information. This task becomes even harder when people such as immigrants wish to find information in their country of residence and do not speak the national language very well. In this paper, we present a new health information system that allows users to search for health information using natural language queries composed of multiple languages. We present the technical details of the system and outline the results of a preliminary user study to demonstrate the usability of the system.

Screening for potential prescription opioid misuse in a michigan medicaid population.

BACKGROUND AND OBJECTIVES: The study's objective was to identify patients at risk for ineffective pain management using patient-specific opioid prescription data. METHODS: We conducted a retrospective review of payer opioid prescription data and patient charts from a rural family medicine group with a residency program. Sixty-one patients were identified who had received three or more prescriptions from two or more providers over a 6-month period. We noted the number of prescriptions and providers, type of medication prescribed, evidence of a medication management agreement (MMA), early refills, increasing doses, frequent telephone calls, reports of lost or stolen medication, history of drug abuse, and proper chart documentation. RESULTS: Seventy percent of the patients were female, 35% had a history of drug abuse, 15% had MMAs, and only 55% had accurate office chart documentation. The number of prescriptions in a 6-month period averaged 8.4 (SD=5.5, range 3 to 28). The average number of prescribers was 3.7 (SD=1.8, range 2 to 10). Patients using non-opioid analgesics had 3.2 fewer prescriptions per 6 months and were less likely to have six or more prescriptions (OR=0.24, 95% CI=0.08--0.73) than those on opioids alone. Concurrent use of non-opioid analgesics, escalating opioid dosage, and number of providers best predicted the number of opioid prescriptions. CONCLUSIONS: Payer data was useful in identifying patients who receive larger-than-expected numbers of opioid prescriptions and factors associated with those larger prescription numbers.