The Crystal Structure of the Extracellular 11-heme Cytochrome UndA Reveals a Conserved 10-heme Motif and Defined Binding Site for Soluble Iron Chelates

Members of the genus Shewanella translocate deca- or undeca-heme cytochromes to the external cell surface thus enabling respiration using extracellular minerals and polynuclear Fe(III) chelates. The high resolution structure of the first undeca-heme outer membrane cytochrome, UndA, reveals a crossed heme chain with four potential electron ingress/egress sites arranged within four domains. Sequence and structural alignment of UndA and the deca-heme MtrF reveals the extra heme of UndA is inserted between MtrF hemes 6 and 7. The remaining UndA hemes can be superposed over the heme chain of the decaheme MtrF, suggesting that a ten heme core is conserved between outer membrane cytochromes. The UndA structure has also been crystallographically resolved in complex with substrates, an Fe(III)-nitrilotriacetate dimer or an Fe(III)-citrate trimer. The structural resolution of these UndA-Fe(III)-chelate complexes provides a rationale for previous kinetic measurements on UndA and other outer membrane cytochromes

Development and evaluation of a diagnostic cytokine-release assay for Mycobacterium suricattae infection in meerkats (Suricata suricatta)

CITATION: Clarke, C., et al. 2017. Development and evaluation of a diagnostic cytokine-release assay for mycobacterium suricattae infection in meerkats (Suricata suricatta). BMC Veterinary Research, 13:2, doi:10.1186/s12917-016-0927-x.The original publication is available at http://bmcvetres.biomedcentral.comBackground: Sensitive diagnostic tools are necessary for the detection of Mycobacterium suricattae infection in meerkats (Suricata suricatta) in order to more clearly understand the epidemiology of tuberculosis and the ecological consequences of the disease in this species. We therefore aimed to develop a cytokine release assay to measure antigen-specific cell-mediated immune responses of meerkats. Results: Enzyme-linked immunosorbent assays (ELISAs) were evaluated for the detection of interferon-gamma (IFN-γ) and IFN-γ inducible protein 10 (IP-10) in meerkat plasma. An IP-10 ELISA was selected to measure the release of this cytokine in whole blood in response to Bovigam® PC-HP Stimulating Antigen, a commercial peptide pool of M. bovis antigens. Using this protocol, captive meerkats with no known M. suricattae exposure (n = 10) were tested and results were used to define a diagnostic cut off value (mean plus 2 standard deviations). This IP-10 release assay (IPRA) was then evaluated in free-living meerkats with known M. suricattae exposure, categorized as having either a low, moderate or high risk of infection with this pathogen. In each category, respectively, 24.7%, 27.3% and 82.4% of animals tested IPRA-positive. The odds of an animal testing positive was 14.0 times greater for animals with a high risk of M. suricattae infection compared to animals with a low risk. Conclusion: These results support the use of this assay as a measure of M. suricattae exposure in meerkat populations. Ongoing longitudinal studies aim to evaluate the value of the IPRA as a diagnostic test of M. suricattae infection in individual animals.http://bmcvetres.biomedcentral.com/articles/10.1186/s12917-016-0927-xPublisher's versio

Automated electroencephalographic discontinuity in cooled newborns predicts cerebral MRI and neurodevelopmental outcome

BACKGROUND AND HYPOTHESIS: Prolonged electroencephalographic (EEG) discontinuity has been associated with poor neurodevelopmental outcomes after perinatal asphyxia but its predictive value in the era of therapeutic hypothermia (TH) is unknown. In infants undergoing TH for hypoxic-ischaemic encephalopathy (HIE) prolonged EEG discontinuity is associated with cerebral tissue injury on MRI and adverse neurodevelopmental outcome. METHOD: Retrospective study of term neonates from three UK centres who received TH for perinatal asphyxia, had continuous two channel amplitude-integrated EEG with EEG for a minimum of 48 h, brain MRI within 6 weeks of birth and neurodevelopmental outcome data at a median age of 24 months. Mean discontinuity was calculated using a novel automated algorithm designed for analysis of the raw EEG signal. RESULTS: Of 49 eligible infants, 17 (35%) had MR images predictive of death or severe neurodisability (unfavourable outcome) and 29 (59%) infants had electrographic seizures. In multivariable logistic regression, mean discontinuity at 24 h and 48 h (both p=0.01), and high seizure burden (p=0.05) were associated with severe cerebral tissue injury on MRI. A mean discontinuity >30 s/min-long epoch, had a specificity and positive predictive value of 100%, sensitivity of 71% and a negative predictive value of 88% for unfavourable neurodevelopmental outcome at a 10 µV threshold. CONCLUSIONS: In addition to seizure burden, excessive EEG discontinuity is associated with increased cerebral tissue injury on MRI and is predictive of abnormal neurodevelopmental outcome in infants treated with TH. The high positive predictive value of EEG discontinuity at 24 h may be valuable in selecting newborns with HIE for adjunctive treatments

Draft Genome Sequence of a Preterm Infant-Derived Isolate of <i>Candida parapsilosis</i>

Candida parapsilosis is a human fungal pathogen of increasing incidence and causes invasive candidiasis, notably in preterm or low-birthweight neonates. Here, we present the genome sequence of C. parapsilosis NCYC 4289, a fecal isolate from a preterm male infant.</p

Linking modern pollen accumulation rates to biomass: Quantitative vegetation reconstruction in the western Klamath Mountains, NW California, USA

Quantitative reconstructions of vegetation abundance from sediment-derived pollen systems provide unique insights into past ecological conditions. Recently, the use of pollen accumulation rates (PAR, grains cm−2 year−1) has shown promise as a bioproxy for plant abundance. However, successfully reconstructing region-specific vegetation dynamics using PAR requires that accurate assessments of pollen deposition processes be quantitatively linked to spatially-explicit measures of plant abundance. Our study addressed these methodological challenges. Modern PAR and vegetation data were obtained from seven lakes in the western Klamath Mountains, California. To determine how to best calibrate our PAR-biomass model, we first calculated the spatial area of vegetation where vegetation composition and patterning is recorded by changes in the pollen signal using two metrics. These metrics were an assemblage-level relevant source area of pollen (aRSAP) derived from extended R-value analysis (sensu Sugita, 1993) and a taxon-specific relevant source area of pollen (tRSAP) derived from PAR regression (sensu Jackson, 1990). To the best of our knowledge, aRSAP and tRSAP have not been directly compared. We found that the tRSAP estimated a smaller area for some taxa (e.g. a circular area with a 225 m radius for Pinus) than the aRSAP (a circular area with a 625 m radius). We fit linear models to relate PAR values from modern lake sediments with empirical, distance-weighted estimates of aboveground live biomass (AGLdw) for both the aRSAP and tRSAP distances. In both cases, we found that the PARs of major tree taxa – Pseudotsuga, Pinus, Notholithocarpus, and TCT (Taxodiaceae, Cupressaceae, and Taxaceae families) – were statistically significant and reasonably precise estimators of contemporary AGLdw. However, predictions weighted by the distance defined by aRSAP tended to be more precise. The relative root-mean squared error for the aRSAP biomass estimates was 9% compared to 12% for tRSAP. Our results demonstrate that calibrated PAR-biomass relationships provide a robust method to infer changes in past plant biomass