Regulatory T cells more effectively suppress Th1-induced airway inflammation compared with Th2

Asthma is a syndrome with different inflammatory phenotypes. Animal models have shown that, after sensitization and allergen challenge, Th2 and Th1 cells contribute to the development of allergic airway disease. We have previously demonstrated that naturally occurring regulatory T cells (nTregs) can only marginally suppress Th2-induced airway inflammation and airway hyperresponsiveness. In this study, we investigated nTreg-mediated suppression of Th2-induced and Th1-induced acute allergic airway disease. We demonstrate in vivo that nTregs exert their suppressive potency via cAMP transfer on Th2- and Th1-induced airway disease. A comparison of both phenotypes revealed that, despite similar cAMP transfers, Th1-driven airway hyperresponsiveness and inflammation are more susceptible to nTreg-dependent suppression, suggesting that potential nTreg-based therapeutic strategies might be more effective in patients with predominantly neutrophilic airway inflammation based on deregulated Th1 response.     

Batatins III-VI, glycolipid ester-type dimers from Ipomoea batatas

Batatins III-VI (1-4), glycolipid ester-type dimers, were isolated from the tuberous roots of sweet potato (Ipomoea batatas) using recycle high performance liquid chromatography. Their structures were characterized by means of several high-resolution NMR and mass spectrometry techniques. These compounds are the first examples of ester-type dimers which consist of two units of the heterotetrasaccharide operculinic acid C. Each unit was esterified by a different amount and type of acid residues: (2S)-methylbutanoic, cinnamic, decanoic (capric) and dodecanoic (lauric) acids. Batatins III-VI (1-4) are an example of the presence of a large number of resin glycoside congeners in each morning glory species caused by partial acylation of their constitutive saccharide cores.     

MPK6, sphinganine and the LCB2a gene from serine palmitoyltransferase are required in the signaling pathway that mediates cell death induced by long chain bases in Arabidopsis

Long chain bases (LCBs) are sphingolipid intermediates acting as second messengers in programmed cell death (PCD) in plants. Most of the molecular and cellular features of this signaling function remain unknown. We induced PCD conditions in Arabidopsis thaliana seedlings and analyzed LCB accumulation kinetics, cell ultrastructure and phenotypes in serine palmitoyltransferase (spt), mitogen-activated protein kinase (mpk), mitogen-activated protein phosphatase (mkp1) and lcb-hydroxylase (sbh) mutants. The lcb2a-1 mutant was unable to mount an effective PCD in response to fumonisin B1 (FB1), revealing that the LCB2a gene is essential for the induction of PCD. The accumulation kinetics of LCBs in wild-type (WT) and lcb2a-1 plants and reconstitution experiments with sphinganine indicated that this LCB was primarily responsible for PCD elicitation. The resistance of the null mpk6 mutant to manifest PCD on FB1 and sphinganine addition and the failure to show resistance on pathogen infection and MPK6 activation by FB1 and LCBs indicated that MPK6 mediates PCD downstream of LCBs. This work describes MPK6 as a novel transducer in the pathway leading to LCB-induced PCD in Arabidopsis, and reveals that sphinganine and the LCB2a gene are required in a PCD process that operates as one of the more effective strategies used as defense against pathogens in plants.     

Engineering homooligomeric proteins to detect weak intersite allosteric communication: aminotransferases, a case study

The existence of low levels of intersubunit communication in homooligomeric enzymes is often difficult to discover, as the identical active sites cannot be probed individually to dissect their interdependent contributions. The homodimeric paralogs, E. coli aspartate- (AATase) and tyrosine aminotransferase (TATase), have not been demonstrated to show allostery. To address this question, we engineered a hybrid aminotransferase containing two distinct catalytic pockets: an AATase and a TATase site. The TATase/AATase hybrid was constructed by grafting an engineered TATase active site into one of the catalytic pockets of E. coli AATase. Each active site conserves its specific catalytic and inhibitor binding properties, and the hybrid catalyzes simultaneously each aminotransferase reaction at the respective site. Importantly, association of a selective inhibitor into one of the catalytic pockets decreases the activity of the second active site by up to 25%, thus proving unequivocally the existence of allosteric communication between active sites. The procedure may be applicable to other homologous sets of enzymes.     

A large-scale forest fragmentation experiment: the Stability of Altered Forest Ecosystems Project

Opportunities to conduct large-scale field experiments are rare, but provide a unique opportunity to reveal the complex processes that operate within natural ecosystems. Here, we review the design of existing, large-scale forest fragmentation experiments. Based on this review, we develop a design for the Stability of Altered Forest Ecosystems (SAFE) Project, a new forest fragmentation experiment to be located in the lowland tropical forests of Borneo (Sabah, Malaysia). The SAFE Project represents an advance on existing experiments in that it: (i) allows discrimination of the effects of landscape-level forest cover from patch-level processes; (ii) is designed to facilitate the unification of a wide range of data types on ecological patterns and processes that operate over a wide range of spatial scales; (iii) has greater replication than existing experiments; (iv) incorporates an experimental manipulation of riparian corridors; and (v) embeds the experimentally fragmented landscape within a wider gradient of land-use intensity than do existing projects. The SAFE Project represents an opportunity for ecologists across disciplines to participate in a large initiative designed to generate a broad understanding of the ecological impacts of tropical forest modification.     

An evaluation of exposure metrics in an epidemiologic study on radio and television broadcast transmitters and the risk of childhood leukemia

Electric field strength values calculated by wave propagation modeling were applied as an exposure metric in a case–control study conducted in Germany to investigate a possible association between radio frequency electromagnetic fields (RF‐EMF) emitted from television and radio broadcast transmitters and the risk of childhood leukemia. To validate this approach it was examined at 850 measurement sites whether calculated RF‐EMF are an improvement to an exposure proxy based on distance from the place of residence to a transmitter. Further, the agreement between measured and calculated RF‐EMF was explored. For dichotomization at the 90% quantiles of the exposure distributions it was found that distance agreed less with measured RF‐EMF (Kappa coefficient: 0.55) than did calculated RF‐EMF (Kappa coefficient: 0.74). Distance was a good exposure proxy for a single transmitter only which uses the frequency bands of amplitude modulated radio, whereas it appeared to be of limited informative value in studies involving several transmitters, particularly if these are operating in different frequency bands. The analysis of the agreement between calculated RF‐EMF and measured RF‐EMF showed a sensitivity of 76.6% and a specificity of 97.4%, leading to an exposure misclassification that still allows one to detect a true odds ratio as low as 1.4 with a statistical power of >80% at a two‐sided significance level of 5% in a study with 2,000 cases and 6,000 controls. Thus, calculated RF‐EMF is confirmed to be an appropriate exposure metric in large‐scale epidemiological studies on broadcast transmitters. Bioelectromagnetics 30:81–91, 2009. © 2008 Wiley‐Liss, Inc.     

The role of N-glycosylation on the enzymatic activity of a Pycnoporus sanguineus laccase

Protein glycosylation, a major post-translational modification, plays essential roles in eukaryotic cells. The glycosylation of fungal laccases has been proposed to be the bottleneck for the heterologous production of the enzyme, so it is important to determine its structure and function. We describe here the detailed N-glycosylation profile of Pycnoporus sanguineus laccase and its influence on some of its enzymatic properties. In this enzyme only high mannose structures were found, being those with 5- and 8-mannose units the most abundant. No other type of sugars was found in contrast to other fungal laccases. Enzymatic cleavage of the N-glycans present in the laccase provoked slight changes in the kinetic parameters, in the thermal stability and in the pH optimum of the enzyme.     

38 The sysytematics of ochromonas (chrysophyceae)

Ochromonas sensu lato is the largest genus in the Chrysophyceae, containing over 100 names. Ochromonas species are biflagellate, naked, plastid‐bearing single cells, distinguished from loricate, scaled, colonial and colorless genera. Most, if not all, species of Ochromonas are mixotrophic, i.e., they photosynthesize but they also engulf bacteria and other small prey. Preliminary evidence from SSU rRNA sequences show that Ochromonas is a polyphyletic genus. Ochromonas tuberculata is the most distinct from all other Ochromonas species. The other Ochromonas species (examined thus far) are scattered in three clades. For example, O. danica and O. sphaerocystis are sister to Poterioochromonas stipitata and P. malhamensis. Four additional species (identified by light microscopy as O. elegans Doflein, O. globosa Skuja, O. ovalis Dolfein, O. sociabilis Pringheim) have SSU rRNA sequences identical to P. malhamensis. Of these, only O. sociabilis has been transferred to Poterioochromonas. Thus, at least some species may be synonymous with others. Two clades of marine species are also known, one containing coastal species and the other containing open ocean species. A number of genera (some also polyphyletic) are interspersed amongst the Ochromonas species (e.g., Chrysolepidomonas, Chrysonephele, Chrysoxys, Cyclonexis, Dinobryon, Epipyxis, Uroglena, Uroglenopsis). The goal of this research (just beginning) is to establish a monophyletic Ochromonas, probably by assigning some species to other genera (existing or new). One major problem is that the type species, O. triangulata Vysotskii, hasn't been observed in over 100 years, and it is unclear which of several clades of Ochromonas contains the type. Results will be discussed.