Bacterial Communities of Frozen Quaternary Sediments of Marine Origin on the Coast of Western Spitsbergen

The bacterial composition of permafrost samples taken during drilling of frozen marine sediments in the area of Barentsburg coal mine on the east coast of Grønfjord Bay of Western Spitsbergen has been studied. The study was based on the analysis of the V4 region of the 16S rRNA gene, carried out using next generation sequencing, as well as using classical microbiological methods (direct luminescence microscopy and aerobic cultivation).The total cell number in permafrost samples ranges from 6.73 ± 0.73 × 106 to 3.37 ± 0.19 107 cells per g. The number of cultivable aerobic bacteria in frozen samples on 1/5 TSA and R2A media ranges from 0 to 6.20 ± 0.45 × 104 CFU/g. Isolates of aerobic bacteria were identified by 16S rRNA gene analysis as representatives of the genera Arthrobacter, Pseudarthrobacter, Psychrobacter, and Rhodoferax. The dominant phyla of the domain Bacteria were Actinobacteria, Proteobacteria, Chloroflexi, Nitrospirae and Firmicutes. As a result of phylogenetic analysis of the dominant operational taxonomic units, representatives of methane oxidizing, sulfate reducing bacteria, as well as heterotrophic bacteria involved in the transformation of organic matter were found

Archaeal Communities of Frozen Quaternary Sediments of Marine Origin on the Coast of Western Spitsbergen

The archaeal composition of permafrost samples taken during the drilling of frozen marine sediments in the area of the Barentsburg coal mine on the east coast of Grønfjord Bay of Western Spitsbergen has been studied. This study is based on an analysis of the V4 region of the 16S rRNA gene, carried out using next-generation sequencing. The general phyla of the Archaea domain are Euryarchaeota, Bathyarchaeota, Thaumarchaeota, and Asgardarchaea. As a result of a phylogenetic analysis of the dominant operational taxonomic units, representatives of methanogenic and methane- and ammonium-oxidizing archaea, as well as heterotrophic archaea, are found. The methanogenic archaea of Euryarchaeota phylum, Methanobacteria class, are found in permafrost with controversial genesis, while the methane-oxidizing archaea of Methanomicrobia class Methanosarcinales order are found in the marine permafrost at Cape Finneset: the ANME-2a, -2b group in layers of 8.6 and 11.7 m and the ANME-2d group (Candidatus Methanoperedens) in a layer of 6.5 m. Ammonium-oxidizing archaea of phylum Thaumarchaeota is present in all types of permafrost, while the order of Nitrososphaerales is found in permafrost with controversial genesis and the order Nitrosopumilales is in permafrost with marine and controversial genesis. Representatives of phylum Bathyarchaeota are found stratigraphically in the most ancient samples under study. Asgardarchaeota superfylum is excluded in the layers of permafrost with marine genesis and is represented by the phyla Lokiarchaeota, Thorarchaeota, and an unclassified group belonging to this superphylum. The presence of methane, ethylene, and ethane in the permafrost of the first sea terrace of Cape Finneset at a depth of 11.7 m, as well as the composition of the archaeal community, give us reason to assume that, before freezing, microbiological processes of anaerobic methane oxidation took place in it, probably received from Tertiary rocks. The results of both this and previous works present the Spitsbergen permafrost as a rich archive of genetic information of little-studied prokaryotic groups

Calibration of photomultiplier arrays

A method is described that allows calibration and assessment of the linearity of response of an array of photomultiplier tubes. The method does not require knowledge of the photomultiplier single photoelectron response model and uses science data directly, thus eliminating the need for dedicated data sets. In this manner all photomultiplier working conditions (e.g. temperature, external fields, etc.) are exactly matched between calibration and science acquisitions. This is of particular importance in low background experiments such as ZEPLIN-III, where methods involving the use of external light sources for calibration are severely constrained

Measurement and simulation of the muon-induced neutron yield in lead

A measurement is presented of the neutron production rate in lead by high energy cosmic-ray muons at a depth of 2850 m water equivalent (w.e.) and a mean muon energy of 260 GeV. The measurement exploits the delayed coincidences between muons and the radiative capture of induced neutrons in a highly segmented tonne scale plastic scintillator detector. Detailed Monte Carlo simulations reproduce well the measured capture times and multiplicities and, within the dynamic range of the instrumentation, the spectrum of energy deposits. By comparing measurements with simulations of neutron capture rates a neutron yield in lead of (View the MathML source) ×10-3 neutrons/muon/(g/cm2) has been obtained. Absolute agreement between simulation and data is of order 25%. Consequences for deep underground rare event searches are discussed

Discriminating lymphomas and reactive lymphadenopathy in lymph node biopsies by gene expression profiling

<p>Abstract</p> <p>Background</p> <p>Diagnostic accuracy of lymphoma, a heterogeneous cancer, is essential for patient management. Several ancillary tests including immunophenotyping, and sometimes cytogenetics and PCR are required to aid histological diagnosis. In this proof of principle study, gene expression microarray was evaluated as a single platform test in the differential diagnosis of common lymphoma subtypes and reactive lymphadenopathy (RL) in lymph node biopsies.</p> <p>Methods</p> <p>116 lymph node biopsies diagnosed as RL, classical Hodgkin lymphoma (cHL), diffuse large B cell lymphoma (DLBCL) or follicular lymphoma (FL) were assayed by mRNA microarray. Three supervised classification strategies (global multi-class, local binary-class and global binary-class classifications) using diagonal linear discriminant analysis was performed on training sets of array data and the classification error rates calculated by leave one out cross-validation. The independent error rate was then evaluated by testing the identified gene classifiers on an independent (test) set of array data.</p> <p>Results</p> <p>The binary classifications provided prediction accuracies, between a subtype of interest and the remaining samples, of 88.5%, 82.8%, 82.8% and 80.0% for FL, cHL, DLBCL, and RL respectively. Identified gene classifiers include LIM domain only-2 (<it>LMO2</it>), Chemokine (C-C motif) ligand 22 (<it>CCL22</it>) and Cyclin-dependent kinase inhibitor-3 (<it>CDK3</it>) specifically for FL, cHL and DLBCL subtypes respectively.</p> <p>Conclusions</p> <p>This study highlights the ability of gene expression profiling to distinguish lymphoma from reactive conditions and classify the major subtypes of lymphoma in a diagnostic setting. A cost-effective single platform "mini-chip" assay could, in principle, be developed to aid the quick diagnosis of lymph node biopsies with the potential to incorporate other pathological entities into such an assay.</p

Genomic Restructuring in the Tasmanian Devil Facial Tumour: Chromosome Painting and Gene Mapping Provide Clues to Evolution of a Transmissible Tumour

Devil facial tumour disease (DFTD) is a fatal, transmissible malignancy that threatens the world's largest marsupial carnivore, the Tasmanian devil, with extinction. First recognised in 1996, DFTD has had a catastrophic effect on wild devil numbers, and intense research efforts to understand and contain the disease have since demonstrated that the tumour is a clonal cell line transmitted by allograft. We used chromosome painting and gene mapping to deconstruct the DFTD karyotype and determine the chromosome and gene rearrangements involved in carcinogenesis. Chromosome painting on three different DFTD tumour strains determined the origins of marker chromosomes and provided a general overview of the rearrangement in DFTD karyotypes. Mapping of 105 BAC clones by fluorescence in situ hybridisation provided a finer level of resolution of genome rearrangements in DFTD strains. Our findings demonstrate that only limited regions of the genome, mainly chromosomes 1 and X, are rearranged in DFTD. Regions rearranged in DFTD are also highly rearranged between different marsupials. Differences between strains are limited, reflecting the unusually stable nature of DFTD. Finally, our detailed maps of both the devil and tumour karyotypes provide a physical framework for future genomic investigations into DFTD

Extensive rewiring of the EGFR network in colorectal cancer cells expressing transforming levels of KRASG13D

Protein-protein-interaction networks (PPINs) organize fundamental biological processes, but how oncogenic mutations impact these interactions and their functions at a network-level scale is poorly understood. Here, we analyze how a common oncogenic KRAS mutation (KRASG13D) affects PPIN structure and function of the Epidermal Growth Factor Receptor (EGFR) network in colorectal cancer (CRC) cells. Mapping >6000 PPIs shows that this network is extensively rewired in cells expressing transforming levels of KRASG13D (mtKRAS). The factors driving PPIN rewiring are multifactorial including changes in protein expression and phosphorylation. Mathematical modelling also suggests that the binding dynamics of low and high affinity KRAS interactors contribute to rewiring. PPIN rewiring substantially alters the composition of protein complexes, signal flow, transcriptional regulation, and cellular phenotype. These changes are validated by targeted and global experimental analysis. Importantly, genetic alterations in the most extensively rewired PPIN nodes occur frequently in CRC and are prognostic of poor patient outcomes.This work was supported by European Union FP7 Grant No. 278568 “PRIMES” and Science Foundation Ireland Investigator Program Grant 14/IA/2395 to W.K. B.K. is supported by SmartNanoTox (Grant no. 686098), NanoCommons (Grant no. 731032), O.R. by MSCA-IF-2016 SAMNets (Grant no. 750688). D.M. is supported by Science Foundation Ireland Career Development award 15-CDA-3495. I.J. is supported by the Canada Research Chair Program (CRC #225404), Krembil Foundation, Ontario Research Fund (GL2-01-030 and #34876), Natural Sciences Research Council (NSERC #203475), Canada Foundation for Innovation (CFI #225404, #30865), and IBM. O.S. is supported by ERC investigator Award ColonCan 311301 and CRUK. I.S. is supported by the Canadian Cancer Society Research Institute (#703889), Genome Canada via Ontario Genomics (#9427 & #9428), Ontario Research fund (ORF/ DIG-501411 & RE08-009), Consortium Québécois sur la Découverte du Médicament (CQDM Quantum Leap) & Brain Canada (Quantum Leap), and CQDM Explore and OCE (#23929). T.C. was supported by a Teagasc Walsh Fellowshi

Induction and processing of the radiation-induced gamma-H2AX signal and Its link to the underlying pattern of DSB: A combined experimental and modelling study

We present here an analysis of DSB induction and processing after irradiation with X-rays in an extended dose range based on the use of the γH2AX assay. The study was performed by quantitative flow cytometry measurements, since the use of foci counting would result in reasonable accuracy only in a limited dose range of a few Gy. The experimental data are complemented by a theoretical analysis based on the GLOBLE model. In fact, original aim of the study was to test GLOBLE predictions against new experimental data, in order to contribute to the validation of the model. Specifically, the γH2AX signal kinetics has been investigated up to 24 h after exposure to increasing photon doses between 2 and 500 Gy. The prolonged persistence of the signal at high doses strongly suggests dose dependence in DSB processing after low LET irradiation. Importantly, in the framework of our modelling analysis, this is related to a gradually increased fraction of DSB clustering at the micrometre scale. The parallel study of γH2AX dose response curves shows the onset of a pronounced saturation in two cell lines at a dose of about 20 Gy. This dose is much lower than expected according to model predictions based on the values usually adopted for the DSB induction yield (≈ 30 DSB/Gy) and for the γH2AX foci extension of approximately 2 Mbp around the DSB. We show and discuss how theoretical predictions and experimental findings can be in principle reconciled by combining an increased DSB induction yield with the assumption of a larger genomic extension for the single phosphorylated regions. As an alternative approach, we also considered in our model the possibility of a 3D spreading-mechanism of the H2AX phosphorylation around the induced DSB, and applied it to the analysis of both the aspects considered. Our results are found to be supportive for the basic assumptions on which GLOBLE is built. Apart from giving new insights into the H2AX phosphorylation process, experiments performed at high doses are of relevance in the context of radiation therapy, where hypo-fractionated schemes become increasingly popular