12 research outputs found
Gene probing reveals the widespread distribution, diversity and abundance of isoprene-degrading bacteria in the environment
Background: Approximately 500 Tg of isoprene are emitted to the atmosphere annually, an amount similar to that of methane, and despite its significant effects on the climate, very little is known about the biological degradation of isoprene in the environment. Isolation and characterisation of isoprene degraders at the molecular level has allowed the development of probes targeting isoA encoding the α-subunit of the isoprene monooxygenase. This enzyme belongs to the soluble diiron centre monooxygenase family and catalyses the first step in the isoprene degradation pathway. The use of probes targeting key metabolic genes is a successful approach in molecular ecology to study specific groups of bacteria in complex environments. Here, we developed and tested a novel isoA PCR primer set to study the distribution, abundance, and diversity of isoprene degraders in a wide range of environments. Results: The new isoA probes specifically amplified isoA genes from taxonomically diverse isoprene-degrading bacteria including members of the genera Rhodococcus, Variovorax, and Sphingopyxis. There was no cross-reactivity with genes encoding related oxygenases from non-isoprene degraders. Sequencing of isoA amplicons from DNA extracted from environmental samples enriched with isoprene revealed that most environments tested harboured a considerable variety of isoA sequences, with poplar leaf enrichments containing more phylogenetically diverse isoA genes. Quantification by qPCR using these isoA probes revealed that isoprene degraders are widespread in the phyllosphere, terrestrial, freshwater and marine environments. Specifically, soils in the vicinity of high isoprene-emitting trees contained the highest number of isoprene-degrading bacteria. Conclusion: This study provides the molecular ecology tools to broaden our knowledge of the distribution, abundance and diversity of isoprene degraders in the environment, which is a fundamental step necessary to assess the impact that microbes have in mitigating the effects of this important climate-active gas
Novel insights into the mechanism of retinol binding by cellular retinol-binding protein
Au-Nanomaterials as a Superior Choice for Near-Infrared Photothermal Therapy
Photothermal therapy (PPT) is a platform to fight cancer by using multiplexed interactive plasmonic nanomaterials as probes in combination with the excellent therapeutic performance of near-infrared (NIR) light. With recent rapid developments in optics and nanotechnology, plasmonic materials have potential in cancer diagnosis and treatment, but there are some concerns regarding their clinical use. The primary concerns include the design of plasmonic nanomaterials which are taken up by the tissues, perform their function and then clear out from the body. Gold nanoparticles (Au NPs) can be developed in different morphologies and functionalized to assist the photothermal therapy in a way that they have clinical value. This review outlines the diverse Au morphologies, their distinctive characteristics, concerns and limitations to provide an idea of the requirements in the field of NIR-based therapeutics
In-Tip Lanthanum Oxide Monolith for the Enrichment of Phosphorylated Biomolecules
Polymeric monoliths
fabricated in tips with embedded materials
of choice are important in separation science. Polymeric backbone
however interferes in the enrichment and thus affects efficiency.
This work focuses on the in-tip fabrication of lanthanum oxide porous
monolith and its application in the enrichment of phosphorylated peptides
and lipids. Polycondensation reaction uses an aqueous solution of
LaCl<sub>3</sub>·7H<sub>2</sub>O with <i>N</i>-methyl
formamide as porogen and propylene oxide as initiator. The aging time
of monolith and temperature condition for the reaction are optimized
to attain porous monolithic tip. A comparison of (i) solid phase batch
extraction using La<sub>2</sub>O<sub>3</sub>, (ii) La<sub>2</sub>O<sub>3</sub> embedded in polyÂ(glycidyl methacrylate (GMA)/divinylbenzene
(DVB)) tip, and (iii) pure La<sub>2</sub>O<sub>3</sub> monolithic
tip shows improved enrichment efficiency in the case of pure La<sub>2</sub>O<sub>3</sub> monolithic tip. The monolithic tip achieves
selectivity of 1:4500 as compared to solid phase extraction (SPE)(1:3500)
and limit of detection down to 0.25 fmol. The in-tip La<sub>2</sub>O<sub>3</sub> monolith strategy has better batch to batch reproducibility,
reduced time of enrichment, and ease of operation in comparison to
solid phase batch extraction. The developed strategy enriches phospho-
content from biological samples like phosvitin and lipovitellin from
egg yolk and phospholipids/phosphopeptides from human serum. The enriched
phospho- moieties are analyzed by matrix-assisted laser desorption
ionization-mass spectrometry (MALDI-MS) except the phospholipids where
laser desorption ionization (LDI)-MS is employed
Newly Fabricated Magnetic Lanthanide Oxides Core–Shell Nanoparticles in Phosphoproteomics
Metal oxides show
high selectivity and sensitivity toward mass
spectrometry based enrichment strategies. Phosphopeptides/phosphoproteins
enrichment from biological samples is cumbersome because of their
low abundance. Phosphopeptides are of interest in enzymes and phosphorylation
pathways which lead to the clinical links of a disease. Magnetic core–shell
lanthanide oxide nanoparticles (Fe<sub>3</sub>O<sub>4</sub>@SiO<sub>2</sub>–La<sub>2</sub>O<sub>3</sub> and Fe<sub>3</sub>O<sub>4</sub>@SiO<sub>2</sub>–Sm<sub>2</sub>O<sub>3</sub>) are fabricated,
characterized by SEM, FTIR, and EDX and employed in the enrichment
of phosphopeptides. The nanoparticles enrich phosphopeptides from
casein variants, nonfat milk, egg yolk, human serum and HeLa cell
extract. The materials and enrichment protocols are designed in a
way that there are almost no nonspecific bindings. The selectivity
is achieved up to 1:8500 using β-casein/BSA mixture and sensitivity
down to 1 atto-mole. Batch-to-batch reproducibility is high with the
reuse of core–shell nanoparticles up to four cycles. The enrichment
followed by MALDI-MS analyses is carried out for the identification
of phosphopeptides from serum digest and HeLa cell extract. Characteristic
phosphopeptides of phosphoproteins are identified from human serum
after the enrichment, which have the diagnostic potential toward prostate
cancer. Thus, the lanthanide based magnetic core–shell materials
offer a highly selective and sensitive workflow in phosphoproteomics
Erratum to: COordination of Standards in MetabOlomicS (COSMOS): facilitating integrated metabolomics data access
Metabolomics has become a crucial phenotyping technique in a range of research fields including medicine, the life sciences, biotechnology and the environmental sciences. This necessitates the transfer of experimental information between research groups, as well as potentially to publishers and funders. After the initial efforts of the metabolomics standards initiative, minimum reporting standards were proposed which included the concepts for metabolomics databases. Built by the community, standards and infrastructure for metabolomics are still needed to allow storage, exchange, comparison and re-utilization of metabolomics data. The Framework Programme 7 EU Initiative ‘coordination of standards in metabolomics’ (COSMOS) is developing a robust data infrastructure and exchange standards for metabolomics data and metadata. This is to support workflows for a broad range of metabolomics applications within the European metabolomics community and the wider metabolomics and biomedical communities’ participation. Here we announce our concepts and efforts asking for re-engagement of the metabolomics community, academics and industry, journal publishers, software and hardware vendors, as well as those interested in standardisation worldwide (addressing missing metabolomics ontologies, complex-metadata capturing and XML based open source data exchange format), to join and work towards updating and implementing metabolomics standards
Automated solvent artifact removal and base plane correction of multidimensional NMR protein spectra by AUREMOL-SSA
Strong solvent signals lead to a disappearance of weak protein signals close to the solvent resonance frequency and to base plane variations all over the spectrum. AUREMOL-SSA provides an automated approach for solvent artifact removal from multidimensional NMR protein spectra. Its core algorithm is based on singular spectrum analysis (SSA) in the time domain and is combined with an automated base plane correction in the frequency domain. The performance of the method has been tested on synthetic and experimental spectra including two-dimensional NOESY and TOCSY spectra and a three-dimensional 1H,13C-HCCH-TOCSY spectrum. It can also be applied to frequency domain spectra since an optional inverse Fourier transformation is included in the algorithm.Bundesministerium für Forschung (BMBF)Deutsche Forschungsgemeinschaft (DFG)European UnionFonds der Chemischen Industrie (FCI