Activation of the pyrin inflammasome by intracellular Burkholderia cenocepacia

Burkholderia cenocepacia is an opportunistic pathogen that causes chronic infection and induces progressive respiratory inflammation in cystic fibrosis patients. Recognition of bacteria by mononuclear cells generally results in the activation of caspase-1 and processing of IL-1β, a major proinflammatory cytokine. In this study, we report that human pyrin is required to detect intracellular B. cenocepacia leading to IL-1β processing and release. This inflammatory response involves the host adapter molecule ASC and the bacterial type VI secretion system (T6SS). Human monocytes and THP-1 cells stably expressing either small interfering RNA against pyrin or YFP-pyrin and ASC (YFP-ASC) were infected with B. cenocepacia and analyzed for inflammasome activation. B. cenocepacia efficiently activates the inflammasome and IL-1β release in monocytes and THP-1. Suppression of pyrin levels in monocytes and THP-1 cells reduced caspase-1 activation and IL-1β release in response to B. cenocepacia challenge. In contrast, overexpression of pyrin or ASC induced a robust IL-1β response to B. cenocepacia, which correlated with enhanced host cell death. Inflammasome activation was significantly reduced in cells infected with T6SS-defective mutants of B. cenocepacia, suggesting that the inflammatory reaction is likely induced by an as yet uncharacterized effector(s) of the T6SS. Together, we show for the first time, to our knowledge, that in human mononuclear cells infected with B. cenocepacia, pyrin associates with caspase-1 and ASC forming an inflammasome that upregulates mononuclear cell IL-1β processing and release.     

Stochastic multi-attribute analysis (SMAA) as an interpretation method for comparative life-cycle assessment (LCA)

Purpose

Comparative life-cycle assessments (LCAs) today lack robust methods of interpretation that help decision makers understand and identify tradeoffs in the selection process. Truncating the analysis at characterization is misleading and existing practices for normalization and weighting may unwittingly oversimplify important aspects of a comparison. This paper introduces a novel approach based on a multi-criteria decision analytic method known as stochastic multi-attribute analysis for life-cycle impact assessment (SMAA-LCIA) that uses internal normalization by means of outranking and exploration of feasible weight spaces.

Methods

To contrast different valuation methods, this study performs a comparative LCA of liquid and powder laundry detergents using three approaches to normalization and weighting: (1) characterization with internal normalization and equal weighting, (2) typical valuation consisting of external normalization and weights, and (3) SMAA-LCIA using outranking normalization and stochastic weighting. Characterized results are often represented by LCA software with respect to their relative impacts normalized to 100 %. Typical valuation approaches rely on normalization references, single value weights, and utilizes discrete numbers throughout the calculation process to generate single scores. Alternatively, SMAA-LCIA is capable of exploring high uncertainty in the input parameters, normalizes internally by pair-wise comparisons (outranking) and allows for the stochastic exploration of weights. SMAA-LCIA yields probabilistic, rather than discrete comparisons that reflect uncertainty in the relative performance of alternatives.

Results and discussion

All methods favored liquid over powder detergent. However, each method results in different conclusions regarding the environmental tradeoffs. Graphical outputs at characterization of comparative assessments portray results in a way that is insensitive to magnitude and thus can be easily misinterpreted. Typical valuation generates results that are oversimplified and unintentionally biased towards a few impact categories due to the use of normalization references. Alternatively, SMAA-LCIA avoids the bias introduced by external normalization references, includes uncertainty in the performance of alternatives and weights, and focuses the analysis on identifying the mutual differences most important to the eventual rank ordering.

Conclusions

SMAA-LCIA is particularly appropriate for comparative LCAs because it evaluates mutual differences and weights stochastically. This allows for tradeoff identification and the ability to sample multiple perspectives simultaneously. SMAA-LCIA is a robust tool that can improve understanding of comparative LCA by decision or policy makers.     

Epidemiology of Visceral Leishmaniasis in Georgia

This study investigated the transmission and prevalence of Leishmania parasite infection of humans in two foci of Visceral Leishmaniasis (VL) in Georgia, the well known focus in Tbilisi in the East, and in Kutaisi, a new focus in the West of the country. The seroprevalence of canine leishmaniasis was investigated in order to understand the zoonotic transmission. Blood samples of 1575 dogs (stray and pet) and 77 wild canids were tested for VL by Kalazar Detect rK39 rapid diagnostic tests. Three districts were investigated in Tbilisi and one in Kutaisi. The highest proportions of seropositive pet dogs were present in District #2 (28.1%, 82/292) and District #1 (26.9%, 24/89) in Tbilisi, compared to 17.3% (26/150) of pet dogs in Kutaisi. The percentage of seropositive stray dogs was also twice as high in Tbilisi (16.1%, n = 670) than in Kutaisi (8%, n = 50); only 2/58 wild animals screened were seropositive (2. 6%). A total of 873 Phlebotomine sand flies were collected, with 5 different species identified in Tbilisi and 3 species in Kutaisi; 2.3% of the females were positive for Leishmania parasites. The Leishmanin Skin Test (LST) was performed on 981 human subjects in VL foci in urban areas in Tbilisi and Kutaisi. A particularly high prevalence of LST positives was observed in Tbilisi District #1 (22.2%, 37.5% and 19.5% for ages 5–9, 15–24 and 25–59, respectively); lower prevalence was observed in Kutaisi (0%, 3.2% and 5.2%, respectively; P<0.05). This study shows that Tbilisi is an active focus for leishmaniasis and that the infection prevalence is very high in dogs and in humans. Although exposure is as yet not as high in Kutaisi, this is a new VL focus. The overall situation in the country is alarming and new control measures are urgently needed.     

Land‐atmosphere exchange of methane from soil thawing to soil freezing in a high‐Arctic wet tundra ecosystem

The land‐atmosphere exchange of methane (CH₄) and carbon dioxide (CO₂) in a high‐Arctic wet tundra ecosystem (Rylekærene) in Zackenberg, north‐eastern Greenland, was studied over the full growing season and until early winter in 2008 and from before snow melt until early winter in 2009. The eddy covariance technique was used to estimate CO₂ fluxes and a combination of the gradient and eddy covariance methods was used to estimate CH₄ fluxes. Small CH₄ bursts were observed during spring thawing 2009, but these existed during short periods and would not have any significant effect on the annual budget. Growing season CH₄ fluxes were well correlated with soil temperature, gross primary production, and active layer thickness. The CH₄ fluxes remained low during the entire autumn, and until early winter. No increase in CH₄ fluxes were seen as the soil started to freeze. However, in autumn 2008 there were two CH₄ burst events that were highly correlated with atmospheric turbulence. They were likely associated with the release of stored CH₄ from soil and vegetation cavities. Over the measurement period, 7.6 and 6.5 g C m^?2 was emitted as CH₄ in 2008 and in 2009, respectively. Rylekærene acted as a C source during the warmer and wetter measurement period 2008, whereas it was a C sink for the colder and drier period of 2009. Wet tundra ecosystems, such as Rylekærene may thus play a more significant role for the climate in the future, as temperature and precipitation are predicted to increase in the high‐Arctic.     

Estimating the potential for coral adaptation to global warming across the Indo‐West Pacific

The potential of reef‐building corals to adapt to increasing sea‐surface temperatures is often debated but has rarely been comprehensively modeled on a region‐wide scale. We used individual‐based simulations to model adaptation to warming in a coral metapopulation comprising 680 reefs and representing the whole of the Central Indo‐West Pacific. Encouragingly, some reefs—most notably Vietnam, Japan, Taiwan, New Caledonia and the southern half of the Great Barrier Reef—exhibited high capacity for adaptation and, in our model, maintained coral cover even under a rapid “business‐as‐usual” warming scenario throughout the modeled period (200 years). Higher resilience of these reefs was observed under all tested parameter settings except the models prohibiting selection and/or migration during warming. At the same time, the majority of reefs in the region tended to collapse within the first 100 years of warming. The adaptive potential (odds of maintaining high coral cover) of a given reef could be predicted based on two metrics: the reef's present‐day temperature, and the proportion of recruits immigrating from warmer locations. The latter metric explains the most variation in adaptive potential, and significantly correlates with actual coral cover changes observed throughout the region between the 1970s and the early 2000s. These findings will help prioritize coral conservation efforts and plan assisted gene flow interventions to boost the adaptive potential of specific coral populations.     

Information-driven self-organization: the dynamical system approach to autonomous robot behavior

In recent years, information theory has come into the focus of researchers interested in the sensorimotor dynamics of both robots and living beings. One root for these approaches is the idea that living beings are information processing systems and that the optimization of these processes should be an evolutionary advantage. Apart from these more fundamental questions, there is much interest recently in the question how a robot can be equipped with an internal drive for innovation or curiosity that may serve as a drive for an open-ended, self-determined development of the robot. The success of these approaches depends essentially on the choice of a convenient measure for the information. This article studies in some detail the use of the predictive information (PI), also called excess entropy or effective measure complexity, of the sensorimotor process. The PI of a process quantifies the total information of past experience that can be used for predicting future events. However, the application of information theoretic measures in robotics mostly is restricted to the case of a finite, discrete state-action space. This article aims at applying the PI in the dynamical systems approach to robot control. We study linear systems as a first step and derive exact results for the PI together with explicit learning rules for the parameters of the controller. Interestingly, these learning rules are of Hebbian nature and local in the sense that the synaptic update is given by the product of activities available directly at the pertinent synaptic ports. The general findings are exemplified by a number of case studies. In particular, in a two-dimensional system, designed at mimicking embodied systems with latent oscillatory locomotion patterns, it is shown that maximizing the PI means to recognize and amplify the latent modes of the robotic system. This and many other examples show that the learning rules derived from the maximum PI principle are a versatile tool for the self-organization of behavior in complex robotic systems.     

Interactions Outside the Proteinase-binding Loop Contribute Significantly to the Inhibition of Activated Coagulation Factor XII by Its Canonical Inhibitor from Corn

Activated factor XII (FXIIa) is selectively inhibited by corn Hageman factor inhibitor (CHFI) among other plasma proteases. CHFI is considered a canonical serine protease inhibitor that interacts with FXIIa through its protease-binding loop. Here we examined whether the protease-binding loop alone is sufficient for the selective inhibition of serine proteases or whether other regions of a canonical inhibitor are involved. Six CHFI mutants lacking different N- and C-terminal portions were generated. CHFI-234, which lacks the first and fifth disulfide bonds and 11 and 19 amino acid residues at the N and C termini, respectively, exhibited no significant changes in FXIIa inhibition (K_i = 3.2 ± 0.4 nm). CHFI-123, which lacks 34 amino acid residues at the C terminus and the fourth and fifth disulfide bridges, inhibited FXIIa with a K_i of 116 ± 16 nm. To exclude interactions outside the FXIIa active site, a synthetic cyclic peptide was tested. The peptide contained residues 20–45 (Protein Data Bank code 1BEA), and a C29D substitution was included to avoid unwanted disulfide bond formation between unpaired cysteines. Surprisingly, the isolated protease-binding loop failed to inhibit FXIIa but retained partial inhibition of trypsin (K_i = 11.7 ± 1.2 μm) and activated factor XI (K_i = 94 ± 11 μm). Full-length CHFI inhibited trypsin with a K_i of 1.3 ± 0.2 nm and activated factor XI with a K_i of 5.4 ± 0.2 μm. Our results suggest that the protease-binding loop is not sufficient for the interaction between FXIIa and CHFI; other regions of the inhibitor also contribute to specific inhibition.     

M protein (M1) of influenza virus: antigenic analysis and intracellular localization with monoclonal antibodies.

A panel of 16 monoclonal antibodies recognizing M protein (M1) of influenza virus was generated. Competition analyses resulted in localization of 14 monoclonal antibodies to three antigenic sites. Three monoclonal antibodies localized to site 1B recognized a peptide synthesized to M1 (residues 220 to 236) with enzyme-linked immunosorbent assay titers equivalent to or greater than that seen with purified M1; therefore, site 1B is located near the C terminus of M1. Sites 2 and 3 localize to the N-terminal half of M1. Antigenic variation of M proteins was seen when the monoclonal antibodies were tested against 14 strains of type A influenza viruses. Several monoclonal antibodies showed specific recognition of A/PR/8/34 and A/USSR/90/77 M proteins and little or no reactivity for all other strains tested. Immunofluorescence analysis with the monoclonal antibodies showed migration of M protein to the nucleus during the replicative cycle and demonstrated association of M protein with actin filaments in the cytoplasm. Use of a vaccinia virus recombinant containing the M-protein gene demonstrated migration of M protein to the nucleus in the absence of synthesis of gene products from other influenza virus RNA segments.     

Breeding habitat and nest‐site selection by an obligatory “nest‐cleptoparasite”, the Amur Falcon Falco amurensis

The selection of a nest site is crucial for successful reproduction of birds. Animals which re‐use or occupy nest sites constructed by other species often have limited choice. Little is known about the criteria of nest‐stealing species to choose suitable nesting sites and habitats. Here, we analyze breeding‐site selection of an obligatory “nest‐cleptoparasite”, the Amur Falcon Falco amurensis. We collected data on nest sites at Muraviovka Park in the Russian Far East, where the species breeds exclusively in nests of the Eurasian Magpie Pica pica. We sampled 117 Eurasian Magpie nests, 38 of which were occupied by Amur Falcons. Nest‐specific variables were assessed, and a recently developed habitat classification map was used to derive landscape metrics. We found that Amur Falcons chose a wide range of nesting sites, but significantly preferred nests with a domed roof. Breeding pairs of Eurasian Hobby Falco subbuteo and Eurasian Magpie were often found to breed near the nest in about the same distance as neighboring Amur Falcon pairs. Additionally, the occurrence of the species was positively associated with bare soil cover, forest cover, and shrub patches within their home range and negatively with the distance to wetlands. Areas of wetlands and fallow land might be used for foraging since Amur Falcons mostly depend on an insect diet. Additionally, we found that rarely burned habitats were preferred. Overall, the effect of landscape variables on the choice of actual nest sites appeared to be rather small. We used different classification methods to predict the probability of occurrence, of which the Random forest method showed the highest accuracy. The areas determined as suitable habitat showed a high concordance with the actual nest locations. We conclude that Amur Falcons prefer to occupy newly built (domed) nests to ensure high nest quality, as well as nests surrounded by available feeding habitats.     

吉林省发现长尾鼠耳蝠

在吉林省通化市采集到长尾鼠耳蝠(Myotis frater)样本9只,为吉林省蝙蝠科新纪录,鉴定为长尾亚种M.f.longicaudatus.本文给出了该蝙蝠的特征描述和相关测量数据,并与文献记录进行了比较.