Biosynthesis of the proteasome inhibitor syringolin A: the ureido group joining two amino acids originates from bicarbonate

<p>Abstract</p> <p>Background</p> <p>Syringolin A, an important virulence factor in the interaction of the phytopathogenic bacterium <it>Pseudomonas syringae </it>pv. <it>syringae </it>B728a with its host plant <it>Phaseolus vulgaris </it>(bean), was recently shown to irreversibly inhibit eukaryotic proteasomes by a novel mechanism. Syringolin A is synthesized by a mixed non-ribosomal peptide synthetase/polyketide synthetase and consists of a tripeptide part including a twelve-membered ring with an N-terminal valine that is joined to a second valine via a very unusual ureido group. Analysis of sequence and architecture of the syringolin A synthetase gene cluster with the five open reading frames <it>sylA-sylE </it>allowed to formulate a biosynthesis model that explained all structural features of the tripeptide part of syringolin A but left the biosynthesis of the unusual ureido group unaccounted for.</p> <p>Results</p> <p>We have cloned a 22 kb genomic fragment containing the <it>sylA-sylE </it>gene cluster but no other complete gene into the broad host range cosmid pLAFR3. Transfer of the recombinant cosmid into <it>Pseudomonas putida </it>and <it>P. syringae </it>pv. <it>syringae </it>SM was sufficient to direct the biosynthesis of <it>bona fide </it>syringolin A in these heterologous organisms whose genomes do not contain homologous genes. NMR analysis of syringolin A isolated from cultures grown in the presence of NaH¹³CO₃revealed preferential ¹³C-labeling at the ureido carbonyl position.</p> <p>Conclusion</p> <p>The results show that no additional syringolin A-specific genes were needed for the biosynthesis of the enigmatic ureido group joining two amino acids. They reveal the source of the ureido carbonyl group to be bicarbonate/carbon dioxide, which we hypothesize is incorporated by carbamylation of valine mediated by the <it>sylC </it>gene product(s). A similar mechanism may also play a role in the biosynthesis of other ureido-group-containing NRPS products known largely from cyanobacteria.</p

A New Species of Boubou (Malaconotidae: Laniarius) from the Albertine Rift

We describe Laniarius willardi, a new species of boubou shrike (Malaconotidae) from the Albertine Rift of Africa. The most conspicuous, distinguishing morphological feature of the species is a gray to blue-gray iris. This and external morphometric data indicate that L. willardi is diagnosable from other black or sooty boubous. Further, L. willardi is genetically diagnosable, and its closest relative is the Mountain Sooty Boubou (L. poensis camerunensis) from Cameroon. The Crimson-breasted Bush-shrike (L. atrococcineus) and the Lowland Sooty Boubou (L. leucorhynchus) are together the sister clade to L. willardi—L.p. camerunensis. Laniarius willardi and the geographically codistributed L. p. holomelas differ by 11.5% in uncorrected sequence divergence, and elevational data taken from museum specimens suggest the possibility of elevational segregation of the species at ∼2,000 m, withL. willardi occurring at lower elevations. Our broad sampling of black and sooty boubou taxa indicate that (1) races of Mountain Sooty Boubou (L. poensis) do not form a monophyletic clade; (2) L. p. camerunensismay represent multiple, nonsister lineages; and (3) at least one race of Fülleborn\u27s Black Boubou (L. fuelleborni usambaricus) is genetically distinct from other races of that species

Solstice: An Electronic Journal of Geography and Mathematics, Volume XXII, Number 2.

This document was delivered over the internet. Look for animations and .kmz files for Google Earth as attachments within the pdf version of this journal. These appear naturally within the online html version. See the full html files, also located at: http://www.imagenet.org/The purpose of Solstice is to promote interaction between geography and mathematics. Articles in which elements of one discipline are used to shed light on the other are particularly sought. Also welcome, are original contributions that are purely geographical or purely mathematical. These may be prefaced (by editor or author) with commentary suggesting directions that might lead toward the desired interaction. Contributed articles will be refereed by geographers and/or mathematicians. Invited articles will be screened by suitable members of the editorial board. IMaGe is open to having authors suggest, and furnish material for, new regular features.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/1/SolsticeDecember2011.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/3/Koenig.pptxhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/4/Arlinghaus2011.pptxhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/5/BridgeOdyssey.kmzhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/6/BridgeOdysseyBlog.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/7/VarroaGlobal.kmzhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/8/Sammataro2011.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/9/AMOVirtualCemetery.kmzhttp://deepblue.lib.umich.edu/bitstream/2027.42/88213/10/ToblerLambertReview.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/88213/17/ArlinghausKoenig.kmzhttps://deepblue.lib.umich.edu/bitstream/2027.42/88213/18/SolsticeVolXXIINo2.pdf8Description of SolsticeDecember2011.pdf : Solstice, December 2011, journal coverDescription of Koenig.pptx : Contemporary Views Along the First Transcontinental Railroad, KoenigDescription of Arlinghaus2011.pptx : The Perimeter Project, Part 6. Arlinghaus and ArlinghausDescription of BridgeOdyssey.kmz : Bridge Odyssey of Craig Robinson, Google Earth file of S. ArlinghausDescription of BridgeOdysseyBlog.pdf : Bridge Odyssey, Blog. Robinson.Description of ArlinghausKoenig.kmz : 3D file with Koenig articleDescription of VarroaGlobal.kmz : Varroa Mite Project of Diana Sammataro. Google Earth file, S. Arlinghaus.Description of Sammataro2011.pdf : Varroa Mite Project, update. D. SammataroDescription of AMOVirtualCemetery.kmz : Virtual Cemetery project with W. E. Arlinghaus. Google Earth file. S. ArlinghausDescription of ToblerLambertReview.pdf : Pdf capture of animated review of Lambert/Tobler documentDescription of SolsticeVolXXIINo2.pdf : Cover of Journa

URBAN TERRAIN CLIMATOLOGY AND REMOTE SENSING *

. Urban areas have been conceived of as monolithic heat islands because traditional ground observation techniques do not lend themselves to more specific analyses. Observations of urban energy-exchange obtained from calibrated electro-optical scanners combined with energy budget simulation techniques provide tools to relate the urban land use mosaic to the heat island phenomenon. Maps of surface energy-related phenomena were made from airborne scanner outputs for selected flightpaths across the city of Baltimore, Maryland. Conditions for the flight time were simulated according to the various types of land use using an energy budget simulation model which lends itself to extrapolation of simulated grid-point conditions into a map form. Maps made by simulation compare sufficiently well with those made by aerial observation to encourage further refinement of the simulation approach.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72392/1/j.1467-8306.1976.tb01110.x.pd

Oxidative stress physiology in relation to life history traits of a free-living vertebrate:the spotted snow skink, Niveoscincus ocellatus

Recent research suggests that oxidative stress, via its links to metabolism and senescence, is a key mechanism linking life history traits such as fecundity and growth with survival; however, this has rarely been put under empirical scrutiny within free-living populations. Using a wild population of live-bearing skinks, we explored how plasma antioxidant activity (OXY), reactive oxidative metabolites (ROM), and the estimated oxidative stress index are associated with female and male life history. We found that male skinks have a significantly higher ROM and estimated oxidative stress index than female skinks, but this was not accompanied by a sex difference in mortality. Both sexes showed a non-linear association between OXY and age, indicating that the oldest and youngest individuals had the lowest OXY. Interestingly, female skinks with high OXY showed a decreased probability of survival to the following season. However, we found no significant associations between female reproductive investment (litter size or litter mass) or parturition date (i.e. metabolism) and oxidative status. Combined, our results offer mixed support for a role of oxidative stress in mediating life history traits and suggest that future studies need to explore oxidative stress during vitellogenesis in addition to using an intra-individual approach to understand the cost of reproduction and patterns of aging.

Using remotely sensed night-time light as a proxy for poverty in Africa

BACKGROUND: Population health is linked closely to poverty. To assess the effectiveness of health interventions it is critical to monitor the spatial and temporal changes in the health indicators of populations and outcomes across varying levels of poverty. Existing measures of poverty based on income, consumption or assets are difficult to compare across geographic settings and are expensive to construct. Remotely sensed data on artificial night time lights (NTL) have been shown to correlate with gross domestic product in developed countries. METHODS: Using national household survey data, principal component analysis was used to compute asset-based poverty indices from aggregated household asset variables at the Administrative 1 level (n = 338) in 37 countries in Africa. Using geographical information systems, mean brightness of and distance to NTL pixels and proportion of area covered by NTL were computed for each Administrative1 polygon. Correlations and agreement of asset-based indices and the three NTL metrics were then examined in both continuous and ordinal forms. RESULTS: At the Administrative 1 level all the NTL metrics distinguished between the most poor and least poor quintiles with greater precision compared to intermediate quintiles. The mean brightness of NTL, however, had the highest correlation coefficient with the asset-based wealth index in continuous (Pearson correlation = 0.64, p &lt; 0.01) and ordinal (Spearman correlation = 0.79, p &lt; 0.01; Kappa = 0.64) forms. CONCLUSION: Metrics of the brightness of NTL data offer a robust and inexpensive alternative to asset-based poverty indices derived from survey data at the Administrative 1 level in Africa. These could be used to explore economic inequity in health outcomes and access to health interventions at sub-national levels where household assets data are not available at the required resolution

Assessing the Quality of Goal Setting in Behavioural Support for Smoking Cessation and its Association with Outcomes

BACKGROUND: Smoking cessation behavioural support can be effective but practitioners differ markedly in effectiveness, possibly due to variation in the quality of delivery of key behaviour change techniques, such as goal setting (i.e. setting a quit date). OBJECTIVES: This study aimed to (i) develop a reliable method for assessing the quality of practitioners' support in setting quit dates and (ii) assess whether quality predicts initiation of abstinence as a first step to quitting. METHODS: A scale for scoring the quality of goal setting was developed from national guidance documents and applied to 85 transcribed behavioural support sessions. Inter-rater reliability was assessed. Associations between quality scores and quit attempts were assessed. RESULTS: The 10-item scale produced had good inter-rater reliability (Kappa = 0.68). Higher quality goal setting was associated with increased self-reported quit attempts (p < .001; OR = 2.60, 95 % CI 1.54-4.40). The scale components 'set a clear quit date' (χ (2) (2, N = 85) = 22.3, p < .001) and 'within an appropriate timeframe' (χ (2) (2, N = 85) = 15.5, p < .001) were independently associated with quit attempts. CONCLUSIONS: It is possible to reliably assess the quality of goal setting in smoking cessation behavioural support. Higher quality of goal setting is associated with greater likelihood of initiating quit attempts

Occasion setters determine responses of putative DA neurons to discriminative stimuli

Midbrain dopamine (DA) neurons are involved in the processing of rewards and reward-predicting stimuli, possibly analogous to reinforcement learning reward prediction errors. Here we studied the activity of putative DA neurons (n=37) recorded in the ventral tegmental area of rats (n=6) performing a behavioural task involving occasion setting. In this task an occasion setter (OS) indicated that the relationship between a discriminative stimulus (DS) and reinforcement is in effect, so that reinforcement of bar pressing occurred only after the OS (tone or houselight) was followed by the DS (houselight or tone). We found that responses of putative DA cells to the DS were enhanced when preceded by the OS, as were behavioural responses to obtain rewards. Surprisingly though, we did not find a homogeneous increase in the mean activity of the population of putative DA neurons to the OS, contrary to predictions of standard temporal-difference models of DA neurons. Instead, putative DA neurons exhibited a heterogeneous response on a single unit level, so that some units increased and others decreased their activity as a response to the OS. Similarly, putative non-DA cells did not show a homogeneous response to the DS on a population level, but also had heterogeneous responses on a single unit level. The heterogeneity in the responses of neurons in the ventral tegmental area may reflect how DA neurons encode context and point to local differences in DA signalling