Aerobic biotransformation of alkyl branched aromatic alkanoic naphthenic acids via two different pathways by a new isolate of Mycobacterium

Naphthenic acids (NAs) are complex mixtures of carboxylic acids found in weathered crude oils and oil sands, and are toxic, corrosive and persistent. However, little is known about the microorganisms and mechanisms involved in NA degradation. We isolated a sediment bacterium (designated strain IS2.3), with 100% 16S rRNA gene sequence identity to Mycobacterium aurum, which degraded synthetic NAs (4'-n-butylphenyl)-4-butanoic acid (n-BPBA) and (4'-t-butylphenyl)-4-butanoic acid (t-BPBA). n-BPBA was readily oxidized with almost complete degradation (96.8% ± 0.3) compared with t-BPBA (77.8% ± 3.7 degraded) by day 49. Cell counts increased fourfold by day 14 but decreased after day 14 for both n- and t-BPBA. At day 14, (4'-butylphenyl)ethanoic acid (BPEA) metabolites were detected. Additional metabolites produced during t-BPBA degradation were identified by mass spectrometry of derivatives as (4'-carboxy-t-butylphenyl)-4-butanoic acid and (4'-carboxy-t-butylphenyl)ethanoic acid; suggesting that strain IS2.3 used omega oxidation of t-BPEA to oxidize the tert-butyl side-chain to produce (4'-carboxy-t-butylphenyl)ethanoic acid, as the primary route for biodegradation. However, strain IS2.3 also produced this metabolite through initial omega oxidation of the tert-butyl side-chain of t-BPBA, followed by beta-oxidation of the alkanoic acid side-chain. In conclusion, an isolate belonging to the genus Mycobacterium degraded highly branched aromatic NAs via two different pathways.     

Aerobic,phenol-induced TCE degradation in completely mixed,continuous-culture reactors

BothPseudomonas putida F1 and a mixed culture were used to study TCE degradation in continuous culture under aerobic, non-methanotrophic conditions. TCE mass balance studies were performed with continuous culture reactors to determine the total percent removed in the reactors, and to quantify the percent removed by air stripping and biodegradation. Adsorption of TCE to biomass was assumed to be negligible. This research demonstrated the feasibility of treating TCE-contaminated water under aerobic, non-methanotrophic conditions with a mixed-culture, continuous-flow system.Initially glucose and acetate were fed as primary substrates. Pnenol, which has been shown to induce TCE-degrading enzymes, was fed at a much lower concentration (20mg/L). Little degradation of TCE was observed when acetate and glucose were the primary substrates. After omitting glucose and acetate from the feed and increasing the phenol concentration to 50mg/L, TCE biotransformation was observed at a significant level (46%). When the phenol concentration in the feed was increased to 420mg/L, 85% of the incoming TCE was estimated to have been biodegraded. Under the same conditions, phenol utilization by the mixed culture was greater than that ofP. putida F1, and TCE degradation by the mixed culture (85%) exceeded that ofP. putida F1 (55%). The estimated percent-of-TCE biodegraded by the mixed culture was consistently greater than 80% when phenol was fed at 420mg/L. Biodegradation of TCE was also observed in mixed-culture, batch experiments.     

GM-CSF priming drives bone marrow-derived macrophages to a pro-inflammatory pattern and downmodulates PGE2 in response to TLR2 ligands

In response to pathogen recognition by Toll-like receptors (TLRs) on their cell surface, macrophages release lipid mediators and cytokines that are widely distributed throughout the body and play essential roles in host responses. Granulocyte macrophage colony-stimulating factor (GM-CSF) is important for the immune response during infections to improve the clearance of microorganisms. In this study, we examined the release of mediators in response to TLR2 ligands by bone marrow-derived macrophages (BMDMs) primed with GM-CSF. We demonstrated that when stimulated with TLR2 ligands, non-primed BMDMs preferentially produced PGE(2) in greater amounts than LTB(4). However, GM-CSF priming shifted the release of lipid mediators by BMDMs, resulting in a significant decrease of PGE(2) production in response to the same stimuli. The decrease of PGE(2) production from primed BMDMs was accompanied by a decrease in PGE-synthase mRNA expression and an increase in TNF-α and nitric oxide (NO) production. Moreover, some GM-CSF effects were potentiated by the addition of IFN-γ. Using a variety of TLR2 ligands, we established that PGE(2) release by GM-CSF-primed BMDMs was dependent on TLR2 co-receptors (TLR1, TLR6), CD14, MyD88 and the nuclear translocation of NFκB but was not dependent on peroxisome proliferator-activated receptor-γ (PPAR-γ) activation. Indeed, GM-CSF priming enhanced TLR2, TLR4 and MyD88 mRNA expression and phospho-IκBα formation. These findings demonstrate that GM-CSF drives BMDMs to present a profile relevant to the host during infections.     

PYCNOCOCCUS PROVASOLII GEN. ET SP. NOV., A COCCOID PRASINOXANTHIN-CONTAINING PHYTOPLANKTER FROM THE WESTERN NORTH ATLANTIC AND GULF OF MEXICO1

The new genus Pycnococcus Guillard is based on several clones from the western North Atlantic and Gulf of Mexico. The type and only described species, Pycnococcus provasolii Guillard, sp. nov., is typified by clone Ω48-23 from the North Atlantic. Cells of Pycnococcus provasolii are solitary, spherical, 1.5–4.0 μm in diameter, have a resistant cell wall lacking sporopollenin, and have the ultrastructural characteristics of green algae. With the light microscope they are scarcely distinguishable from cells of other coccoid planktonic organisms. In pigmentation P. provasolii resembles Micromonas pusilla, Mantoniella squamata, and Mamiella gilva in having chl a, much chl b, Mg 2,4-divinylphaeoporphyrin a₅ monomethyl ester (presumably), and prasinoxanthin as a major xanthophyll. The pyrenoid of P. provasolii has a cytoplasmic channel, which is unique among species closely related to it. Flagellates, occurring rarely in culture, are similar to but distinguishable from known Pedinomonas species by size and shape. Pycnococcus provasolii is referred to the new family Pycnococcaceae Guillard, in the order Mamiellales of the class Micromonadophyceae (Chlorophyta). Clones of Pycnococcus provasolii are oceanic in nutritional characteristics, require only vitamin B₁₂ in culture, and are well adapted to growth under blue or blue-violet light of low intensity.     

Socioeconomic and Environmental Risk Factors among Rheumatic Heart Disease Patients in Uganda

Background

Although low socioeconomic status, and environmental factors are known risk factors for rheumatic heart disease in other societies, risk factors for rheumatic heart disease remain less well described in Uganda.

Aims and Objective

The objective of this study was to investigate the role of socio-economic and environmental factors in the pathogenesis of rheumatic heart disease in Ugandan patients.

Methods

This was a case control study in which rheumatic heart disease cases and normal controls aged 5–60 years were recruited and investigated for socioeconomic and environmental risk factors such as income status, employment status, distance from the nearest health centre, number of people per house and space area per person.

Results

486 participants (243 cases and 243 controls) took part in the study. Average age was 32.37+/−14.6 years for cases and 35.75+/−12.6 years for controls. At univariate level, Cases tended to be more overcrowded than controls; 8.0+/−3.0 versus 6.0+/−3.0 persons per house. Controls were better spaced at 25.2 square feet versus 16.9 for cases. More controls than cases were employed; 45.3% versus 21.1%. Controls lived closer to health centers than the cases; 4.8+/−3.8 versus 3.3+/−12.9 kilometers. At multivariate level, the odds of rheumatic heart disease was 1.7 times higher for unemployment status (OR = 1.7, 95% CI = 1.05–8.19) and 1.3 times higher for overcrowding (OR = 1.35, 95% CI = 1.1–1.56). There was interaction between overcrowding and longer distance from the nearest health centre (OR = 1.20, 95% CI = 1.05–1.42).

Conclusion

The major findings of this study were that there was a trend towards increased risk of rheumatic heart disease in association with overcrowding and unemployment. There was interaction between overcrowding and distance from the nearest health center, suggesting that the effect of overcrowding on the risk of acquiring rheumatic heart disease increases with every kilometer increase from the nearest health center.     

Multiple sources of evidence for density dependence in the endangered Hawaiian stilt (Himantopus mexicanus knudseni)

Hawaiian stilts (Himantopus mexicanus knudseni) are an endangered subspecies of the Black-necked stilt endemic to the Hawaiian Islands. Despite long-term study, the main drivers of Hawaiian stilt population dynamics are poorly understood. We tested for density dependence using two sources of evidence: a 30-year time series of annual estimated range-wide abundance, and two 15+ year time series of reproductive success. Using separate methods with independent data, sources allowed us to make up for the potentially positive bias of one approach with the more conservative nature of the second. We compared nonlinear density-dependent and density-independent population model fits to our time-series data, using both frequentist and Bayesian state-space approaches. Across both approaches, density-dependent models best fit observed population dynamics, with lower AICc and cross-validation statistics compared to density-independent models. Among density-dependent models, a conditional model in which density-independent dynamics occur below a population size threshold (~850–1,000 birds), and then density-dependent dynamics occur above that threshold, performed best across Bayesian and frequentist model comparisons, with the Ricker model ranked next or equivalently. Our analysis of reproduction data revealed a strong negative effect of local adult density on nest success (proportion of nests hatching at least one chick) at Kealia National Wildlife Refuge on Maui, where few alternative breeding habitats are available, but no such effect at another site where many nearby alternative wetlands are available. These congruent results across independent datasets and analytical approaches support the hypothesis that Hawaiian stilts exhibit density dependence across their range.     

Prime-boost vaccination with rBCG/rAd35 enhances CD8⁺ cytolytic T-cell responses in lesions from Mycobacterium tuberculosis-infected primates

To prevent the global spread of tuberculosis (TB) infection, a novel vaccine that triggers potent and long-lived immunity is urgently required. A plasmid-based vaccine has been developed to enhance activation of major histocompatibility complex (MHC) class I–restricted CD8+ cytolytic T cells using a recombinant Bacille Calmette-Guérin (rBCG) expressing a pore-forming toxin and the Mycobacterium tuberculosis (Mtb) antigens Ag85A, 85B and TB10.4 followed by a booster with a nonreplicating adenovirus 35 (rAd35) vaccine vector encoding the same Mtb antigens. Here, the capacity of the rBCG/rAd35 vaccine to induce protective and biologically relevant CD8⁺ T-cell responses in a nonhuman primate model of TB was investigated. After prime/boost immunizations and challenge with virulent Mtb in rhesus macaques, quantification of immune responses at the single-cell level in cryopreserved tissue specimen from infected organs was performed using in situ computerized image analysis as a technological platform. Significantly elevated levels of CD3⁺ and CD8⁺ T cells as well as cells expressing interleukin (IL)-7, perforin and granulysin were found in TB lung lesions and spleen from rBCG/rAd35-vaccinated animals compared with BCG/rAd35-vaccinated or unvaccinated animals. The local increase in CD8⁺ cytolytic T cells correlated with reduced expression of the Mtb antigen MPT64 and also with prolonged survival after the challenge. Our observations suggest that a protective immune response in rBCG/rAd35-vaccinated nonhuman primates was associated with enhanced MHC class I antigen presentation and activation of CD8+ effector T-cell responses at the local site of infection in Mtb-challenged animals.     

A triple arg motif mediates α(2B)-adrenergic receptor interaction with Sec24C/D and export

Recent studies have demonstrated that cargo exit from the endoplasmic reticulum (ER) may be directed by ER export motifs recognized by components of the coat protein II (COPII) vesicles. However, little is known about ER export motifs and vesicle targeting of the G protein-coupled receptor (GPCR) superfamily. Here, we have demonstrated that a triple Arg (3R) motif in the third intracellular loop functions as a novel ER export signal for α(2B)-adrenergic receptor (α(2B)-AR). The 3R motif mediates α(2B)-AR interaction with Sec24C/D and modulates ER exit, cell surface transport and function of α(2B)-AR. Furthermore, export function of the 3R motif is independent of its position within α(2B)-AR and can be conferred to CD8 glycoprotein. These data provide the first evidence implicating that export of GPCRs is controlled by code-directed interactions with selective components of the COPII transport machinery.