Metaproteomics of complex microbial communities in biogas plants

Production of biogas from agricultural biomass or organic wastes is an important source of renewable energy. Although thousands of biogas plants (BGPs) are operating in Germany, there is still a significant potential to improve yields, e.g. from fibrous substrates. In addition, process stability should be optimized. Besides evaluating technical measures, improving our understanding of microbial communities involved into the biogas process is considered as key issue to achieve both goals. Microscopic and genetic approaches to analyse community composition provide valuable experimental data, but fail to detect presence of enzymes and overall metabolic activity of microbial communities. Therefore, metaproteomics can significantly contribute to elucidate critical steps in the conversion of biomass to methane as it delivers combined functional and phylogenetic data. Although metaproteomics analyses are challenged by sample impurities, sample complexity and redundant protein identification, and are still limited by the availability of genome sequences, recent studies have shown promising results. In the following, the workflow and potential pitfalls for metaproteomics of samples from full-scale BGP are discussed. In addition, the value of metaproteomics to contribute to the further advancement of microbial ecology is evaluated. Finally, synergistic effects expected when metaproteomics is combined with advanced imaging techniques, metagenomics, metatranscriptomics and metabolomics are addressed

A Robust and Universal Metaproteomics Workflow for Research Studies and Routine Diagnostics Within 24 h Using Phenol Extraction, FASP Digest, and the MetaProteomeAnalyzer

The investigation of microbial proteins by mass spectrometry (metaproteomics) is a key technology for simultaneously assessing the taxonomic composition and the functionality of microbial communities in medical, environmental, and biotechnological applications. We present an improved metaproteomics workflow using an updated sample preparation and a new version of the MetaProteomeAnalyzer software for data analysis. High resolution by multidimensional separation (GeLC, MudPIT) was sacrificed to aim at fast analysis of a broad range of different samples in less than 24 h. The improved workflow generated at least two times as many protein identifications than our previous workflow, and a drastic increase of taxonomic and functional annotations. Improvements of all aspects of the workflow, particularly the speed, are first steps toward potential routine clinical diagnostics (i.e., fecal samples) and analysis of technical and environmental samples. The MetaProteomeAnalyzer is provided to the scientific community as a central remote server solution at www.mpa.ovgu.de.Peer Reviewe

Tyrosine phosphorylation of the N-Methyl-D-Aspartate receptor 2B subunit in spinal cord contributes to remifentanil-induced postoperative hyperalgesia: the preventive effect of ketamine

<p>Abstract</p> <p>Background</p> <p>Experimental and clinical studies showed that intraoperative infusionof remifentanil has been associated with postoperative hyperalgesia. Previous reports suggested that spinal N-methyl-D-aspartate (NMDA) receptors may contribute to the development and maintenance of opioid-induced hyperalgesia. In the present study, we used a rat model of postoperative pain to investigate the role of tyrosine phosphorylation of NMDA receptor 2B (NR2B) subunit in spinal cord in the postoperative hyperalgesia induced by remifentanil and the intervention of pretreatment with ketamine.</p> <p>Results</p> <p>Intraoperative infusion of remifentanil (0.04 mg/kg, subcutaneous) significantly enhanced mechanical allodynia and thermal hyperalgesia induced by the plantar incision during the postoperative period (each lasting between 2 h and 48 h), which was attenuated by pretreatment with ketamine (10 mg/kg, subcutaneous). Correlated with the pain behavior changes, immunocytochemical and western blotting experiments in our study revealed that there was a marked increase in NR2B phosphorylation at Tyr1472 in the superficial dorsal horn after intraoperative infusion of remifentanil, which was attenuated by pretreatment with ketamine.</p> <p>Conclusions</p> <p>This study provides direct evidence that tyrosine phosphorylation of the NR2B at Tyr1472 in spinal dosal horn contributes to postoperative hyperalgesia induced by remifentanil and supports the potential therapeutic value of ketamine for improving postoperative hyperalgesia induced by remifentanil.</p

Over-Ocean Validation of the Global Convective Diagnostic

The global convective diagnostic (GCD) is a bispectral (infrared and water vapor), day–night scheme for operationally mapping deep convection by means of geostationary satellite images. This article describes a test of GCD performance over tropical and subtropical waters near North America. The test consists of six cases, each involving a convective cloud complex. A seventh case treats convection over land. For each case, a map of deep convection was constructed from image pairs from Geostationary Operational Environmental Satellite-12 (GOES-12). Case by case and for all maritime cases together, the GCD map was compared with a convective parameter derived from the radar on the Tropical Rainfall Measuring Mission (TRMM), a polar-orbiting satellite. In general, each GCD map showed a bloblike feature. In each case, the radar convective pixels typically fell within the GCD blob. However, (except for the land case) the GCD predicted far too many convective pixels. In the maritime cases overprediction was reduced (without correspondingly impairing other measures of performance) by lowering the nominal GCD threshold. With this adjustment in place, for the six maritime cases taken individually, the GCD tended to yield more consistent results than did a monospectral (infrared) convective scheme. With the cases combined, at the lower threshold the GCD performed somewhat better than one of the more stable versions of the infrared scheme. Comparison with lightning events (also observed by TRMM) suggests the possibility of future improvement to the GCD through the incorporation of geostationary satellite observations of lightning

Status of a DEPFET pixel system for the ILC vertex detector

We have developed a prototype system for the ILC vertex detector based on DEPFET pixels. The system operates a 128x64 matrix (with ~35x25 square micron large pixels) and uses two dedicated microchips, the SWITCHER II chip for matrix steering and the CURO II chip for readout. The system development has been driven by the final ILC requirements which above all demand a detector thinned to 50 micron and a row wise read out with line rates of 20MHz and more. The targeted noise performance for the DEPFET technology is in the range of ENC=100 e-. The functionality of the system has been demonstrated using different radioactive sources in an energy range from 6 to 40keV. In recent test beam experiments using 6GeV electrons, a signal-to-noise ratio of S/N~120 has been achieved with present sensors being 450 micron thick. For improved DEPFET systems using 50 micron thin sensors in future, a signal-to-noise of 40 is expected.Comment: Invited poster at the International Symposium on the Development of Detectors for Particle, AstroParticle and Synchrotron Radiation Experiments, Stanford CA (SNIC06) 6 pages, 12 eps figure

RUNX1/ETO blocks selectin-mediated adhesion via epigenetic silencing of PSGL-1

RUNX1/ETO (RE),the t(8;21)-derived leukemic transcription factor associated with acute myeloid leukemia (AML) development, deregulates genes involved in differentiation, self-renewal and proliferation. In addition, these cells show differences in cellular adhesion behavior whose molecular basis is not well understood. Here, we demonstrate that RE epigenetically silences the gene encoding P-Selectin Glycoprotein Ligand-1 (PSGL-1) and downregulates PSGL-1 expression in human CD34+ and murine lin-hematopoietic progenitor cells. Levels of PSGL-1 inversely and dose-dependently correlate with RE oncogene levels. However, a DNA-binding defective mutant fails to downregulate PSGL-1. We show by ChIP experiments that the PSGL-1 promoter is a direct target of RE and binding is accompanied by high levels of the repressive chromatin mark histone H3K27me3. In t(8;21)+ Kasumi-1 cells, PSGL-1 expression is completely restored at both the mRNA and cell surface protein levels following RE downregulation with short hairpin RNA (shRNA) or RE inhibition with tetramerization-blocking peptides, and at the promoter H3K27me3 is replaced by the activating chromatin mark H3K9ac as well as by RNA polymerase II. Upregulation of PSGL-1 restores the binding of cells to P- and E-selectin and re-establishes myeloid-specific cellular adhesion while it fails to bind to lymphocyte-specific L-selectin. Overall, our data suggest that the RE oncoprotein epigenetically represses PSGL-1 via binding to its promoter region and thus affects the adhesive behavior of t(8;21)+ AML cells

Metaproteome analysis reveals that syntrophy, competition, and phage-host interaction shape microbial communities in biogas plants

Background: In biogas plants, complex microbial communities produce methane and carbon dioxide by anaerobic digestion of biomass. For the characterization of the microbial functional networks, samples of 11 reactors were analyzed using a high-resolution metaproteomics pipeline. Results: Examined methanogenesis archaeal communities were either mixotrophic or strictly hydrogenotrophic in syntrophy with bacterial acetate oxidizers. Mapping of identified metaproteins with process steps described by the Anaerobic Digestion Model 1 confirmed its main assumptions and also proposed some extensions such as syntrophic acetate oxidation or fermentation of alcohols. Results indicate that the microbial communities were shaped by syntrophy as well as competition and phage-host interactions causing cell lysis. For the families Bacillaceae, Enterobacteriaceae, and Clostridiaceae, the number of phages exceeded up to 20-fold the number of host cells. Conclusion: Phage-induced cell lysis might slow down the conversion of substrates to biogas, though, it could support the growth of auxotrophic microbes by cycling of nutrients. © 2019 The Author(s)

Faktor Faktor Yang Berhubungan Dengan Perilaku Deteksi Dini Kanker Leher Rahim Metode Inspeksi Visual Asam Asetat Di Puskesmas Candiroto Kabupaten Temanggung

Perilaku deteksi dini kanker leher rahim dengan metode IVA adalah bentuk tindakan pemeriksaan deteksi dini kanker leher rahim dengan metode IVA. Kesadaran perempuan Indonesia untuk melakukan deteksi kanker leher rahim secara teratur masih rendah. Hal ini terlihat dari rendahnya cakupan deteksi dini kanker leher rahim dengan metode Inspeksi Visual Asam Asetat. Untuk tingkat Jawa Tengan hanya 1,08% dari jumlah wanita usia 30-50 tahun, Puskesmas Candiroto sekitar 5%. Tujuan penelitian ini adalah menganalisis faktor yang berhubungan dengan perilaku deteksi dini kanker leher rahim metode Inspeksi Visual Asam Asetat di Puskesmas Candiroto Kabupaten Temanggung. Penelitian ini merupakan penelitian dengan pendekatan cross sectional dengan analisis data uji chi-square dengan taraf signifikansi 95%. Subjek penelitian ini adalah wanita usia subur di wilayakh kerja Puskesmas Candiroto sejumlah 100 orang dengan menggunakan simple random sampling. Hasil uji chi-square ada hubungan antara keterpaparan informasi(p=0,1), dukungan suami (p=0,026), dan dukungan teman (p<0,0001) dengan perilaku deteksi dini kanker leher rahim metode IVA, serta tidak ada hubungan antara umur (p=0,07), pendidikan (p=0,17), pekerjaan (p=0,51), pengetahuan ((p=0,36), sikap (p=0,097), jarak (p=0,1), dukungan tenaga kesehatan (p=0,1) dengan perilaku deteksi dini kanker leher rahim metode IVA. dapat disimpulkan faktor yang berhubungan dengan perilaku deteksi dini kanker leher rahim metode Inspeksi Visual Asam Asetat di Puskesmas Candiroto Kabupaten Temanggung yaitu keterpaparan informasi, dukungan suami, dan dukungan teman. Kata Kunci: kanker leher rahim, deteksi dini, IV