Cloning, sequencing, and mutation of a gene for azurin in Methylobacillus flagellatum KT

The gene cluster for methylamine utilization (mau genes) has been cloned from the obligate methylotrophic bacterium Methylobacillus flagellatum KT. Partial sequence data showed that the organization of these genes was similar to that found in Methylophilus methylotrophus W3A1-NS, including the lack of a gene for amicyanin, which had been thought to be the electron acceptor for methylamine dehydrogenase in M. flagellatum KT. However, a gene encoding azurin was discovered at the 3' end of the mau gene cluster, transcribed in the opposite orientation. A mutant with a defect in this gene showed impaired growth on methylamine, suggesting that azurin is involved in methylamine oxidation in M. flagellatum KT

Clastic matrix in EH3 chondrites

Patches of clastic matrix (15 to 730 μm in size) constitute 4.9 vol% of EH3 Yamato (Y-) 691 and 11.7 vol% of EH3 Allan Hills (ALH) 81189. Individual patches in Y-691 consist of 1) ~25 vol% relatively coarse opaque grain fragments and polycrystalline assemblages of kamacite, schreibersite, perryite, troilite (some grains with daubréelite exsolution lamellae), niningerite, oldhamite, and caswellsilverite; 2) ~30 vol% relatively coarse silicate grains including enstatite, albitic plagioclase, silica and diopside; and 3) an inferred fine nebular component (~45 vol%) comprised of submicrometer-size grains. Clastic matrix patches in ALH 81189 contain relatively coarse grains of opaques (~20 vol%; kamacite, schreibersite, perryite and troilite) and silicates (~30 vol%; enstatite, silica and forsterite) as well as an inferred fine nebular component (~50 vol%). The O-isotopic composition of clastic matrix in Y-691 is indistinguishable from that of olivine and pyroxene grains in adjacent chondrules; both sets of objects lie on the terrestrial mass-fractionation line on the standard three-isotope graph. Some patches of fine-grained matrix in Y-691 have distinguishable bulk concentrations of Na and K, inferred to be inherited from the solar nebula. Some patches in ALH 81189 differ in their bulk concentrations of Ca, Cr, Mn, and Ni. The average compositions of matrix material in Y-691 and ALH 81189 are similar but not identical—matrix in ALH 81189 is much richer in Mn (0.23 ± 0.05 versus 0.07 ± 0.02 wt%) and appreciably richer in Ni (0.36 ± 0.10 versus 0.18 ± 0.05 wt%) than matrix in Y-691. Each of the two whole-rocks exhibits a petrofabric, probably produced by shock processes on their parent asteroid

Biomarkers Associated with Organ-Specific Involvement in Juvenile Systemic Lupus Erythematosus

Juvenile systemic lupus erythematosus (JSLE) is characterised by onset before 18 years of age and more severe disease phenotype, increased morbidity and mortality compared to adult-onset SLE. Management strategies in JSLE rely heavily on evidence derived from adult-onset SLE studies; therefore, identifying biomarkers associated with the disease pathogenesis and reflecting particularities of JSLE clinical phenotype holds promise for better patient management and improved outcomes. This narrative review summarises the evidence related to various traditional and novel biomarkers that have shown a promising role in identifying and predicting specific organ involvement in JSLE and appraises the evidence regarding their clinical utility, focusing in particular on renal biomarkers, while also emphasising the research into cardiovascular, haematological, neurological, skin and joint disease-related JSLE biomarkers, as well as genetic biomarkers with potential clinical applications

Agent-based Modelling of Socio-Ecological Systems: Models, Projects and Ontologies

Socio-Ecological Systems (SESs) are the systems in which our everyday lives are embedded, so understanding them is important. The complex properties of such systems make modelling an indispensable tool for their description and analysis. Human actors play a pivotal role in SESs, but their interactions with each other and their environment are often underrepresented in SES modelling. We argue that more attention should be given to social aspects in models of SESs, but this entails additional kinds of complexity. Modelling choices need to be as transparent as possible, and to be based on analysis of the purposes and limitations of modelling. We recommend thinking in terms of modelling projects rather than single models. Such a project may involve multiple models adopting different modelling methods. We argue that agent-based models (ABMs) are an essential tool in an SES modelling project, but their expressivity, which is their major advantage, also produces problems with model transparency and validation. We propose the use of formal ontologies to make the structure and meaning of models as explicit as possible, facilitating model design, implementation, assessment, comparison and extension

Ex Vivo Treatment with a Novel Synthetic Aminoglycoside NB54 in Primary Fibroblasts from Rett Syndrome Patients Suppresses MECP2 Nonsense Mutations

BACKGROUND: Nonsense mutations in the X-linked methyl CpG-binding protein 2 (MECP2) comprise a significant proportion of causative MECP2 mutations in Rett syndrome (RTT). Naturally occurring aminoglycosides, such as gentamicin, have been shown to enable partial suppression of nonsense mutations related to several human genetic disorders, however, their clinical applicability has been compromised by parallel findings of severe toxic effects. Recently developed synthetic NB aminoglycosides have demonstrated significantly improved effects compared to gentamicin evident in substantially higher suppression and reduced acute toxicity in vitro. RESULTS: We performed comparative study of suppression effects of the novel NB54 and gentamicin on three MECP2 nonsense mutations (R294X, R270X and R168X) common in RTT, using ex vivo treatment of primary fibroblasts from RTT patients harboring these mutations and testing for the C-terminal containing full-length MeCP2. We observed that NB54 induces dose-dependent suppression of MECP2 nonsense mutations more efficiently than gentamicin, which was evident at concentrations as low as 50 µg/ml. NB54 read-through activity was mutation specific, with maximal full-length MeCP2 recovery in R168X (38%), R270X (27%) and R294X (18%). In addition, the recovered MeCP2 was translocated to the cell nucleus and moreover led to parallel increase in one of the most important MeCP2 downstream effectors, the brain derived neurotrophic factor (BDNF). CONCLUSION: Our findings suggest that NB54 may induce restoration of the potentially functional MeCP2 in primary RTT fibroblasts and encourage further studies of NB54 and other rationally designed aminoglycoside derivatives as potential therapeutic agents for nonsense MECP2 mutations in RTT

Stressed out symbiotes:hypotheses for the influence of abiotic stress on arbuscular mycorrhizal fungi

Abiotic stress is a widespread threat to both plant and soil communities. Arbuscular mycorrhizal (AM) fungi can alleviate effects of abiotic stress by improving host plant stress tolerance, but the direct effects of abiotic stress on AM fungi are less well understood. We propose two hypotheses predicting how AM fungi will respond to abiotic stress. The stress exclusion hypothesis predicts that AM fungal abundance and diversity will decrease with persistent abiotic stress. The mycorrhizal stress adaptation hypothesis predicts that AM fungi will evolve in response to abiotic stress to maintain their fitness. We conclude that abiotic stress can have effects on AM fungi independent of the effects on the host plant. AM fungal communities will change in composition in response to abiotic stress, which may mean the loss of important individual species. This could alter feedbacks to the plant community and beyond. AM fungi will adapt to abiotic stress independent of their host plant. The adaptation of AM fungi to abiotic stress should allow the maintenance of the plant-AM fungal mutualism in the face of changing climates. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00442-016-3673-7) contains supplementary material, which is available to authorized users

Application of the bacteriophage Mu-driven system for the integration/amplification of target genes in the chromosomes of engineered Gram-negative bacteria—mini review

The advantages of phage Mu transposition-based systems for the chromosomal editing of plasmid-less strains are reviewed. The cis and trans requirements for Mu phage-mediated transposition, which include the L/R ends of the Mu DNA, the transposition factors MuA and MuB, and the cis/trans functioning of the E element as an enhancer, are presented. Mini-Mu(LR)/(LER) units are Mu derivatives that lack most of the Mu genes but contain the L/R ends or a properly arranged E element in cis to the L/R ends. The dual-component system, which consists of an integrative plasmid with a mini-Mu and an easily eliminated helper plasmid encoding inducible transposition factors, is described in detail as a tool for the integration/amplification of recombinant DNAs. This chromosomal editing method is based on replicative transposition through the formation of a cointegrate that can be resolved in a recombination-dependent manner. (E-plus)- or (E-minus)-helpers that differ in the presence of the trans-acting E element are used to achieve the proper mini-Mu transposition intensity. The systems that have been developed for the construction of stably maintained mini-Mu multi-integrant strains of Escherichia coli and Methylophilus methylotrophus are described. A novel integration/amplification/fixation strategy is proposed for consecutive independent replicative transpositions of different mini-Mu(LER) units with “excisable” E elements in methylotrophic cells