Metabolomic Derangements Are Associated with Mortality in Critically Ill Adult Patients

Objective

To identify metabolomic biomarkers predictive of Intensive Care Unit (ICU) mortality in adults.

Rationale

Comprehensive metabolomic profiling of plasma at ICU admission to identify biomarkers associated with mortality has recently become feasible.

Methods

We performed metabolomic profiling of plasma from 90 ICU subjects enrolled in the BWH Registry of Critical Illness (RoCI). We tested individual metabolites and a Bayesian Network of metabolites for association with 28-day mortality, using logistic regression in R, and the CGBayesNets Package in MATLAB. Both individual metabolites and the network were tested for replication in an independent cohort of 149 adults enrolled in the Community Acquired Pneumonia and Sepsis Outcome Diagnostics (CAPSOD) study.

Results

We tested variable metabolites for association with 28-day mortality. In RoCI, nearly one third of metabolites differed among ICU survivors versus those who died by day 28 (N = 57 metabolites, p<.05). Associations with 28-day mortality replicated for 31 of these metabolites (with p<.05) in the CAPSOD population. Replicating metabolites included lipids (N = 14), amino acids or amino acid breakdown products (N = 12), carbohydrates (N = 1), nucleotides (N = 3), and 1 peptide. Among 31 replicated metabolites, 25 were higher in subjects who progressed to die; all 6 metabolites that are lower in those who die are lipids. We used Bayesian modeling to form a metabolomic network of 7 metabolites associated with death (gamma-glutamylphenylalanine, gamma-glutamyltyrosine, 1-arachidonoylGPC(20:4), taurochenodeoxycholate, 3-(4-hydroxyphenyl) lactate, sucrose, kynurenine). This network achieved a 91% AUC predicting 28-day mortality in RoCI, and 74% of the AUC in CAPSOD (p<.001 in both populations).

Conclusion

Both individual metabolites and a metabolomic network were associated with 28-day mortality in two independent cohorts. Metabolomic profiling represents a valuable new approach for identifying novel biomarkers in critically ill patients.     

Fluctuations of an introduced population of Svalbard reindeer: the effects of density dependence and climatic variation

The relative contribution of density-dependent and density-independent factors on variation in the population growth rate of an introduced population Svalbard reindeer was studied by time series analysis. No significant effects of either direct or delayed density-dependence were found. Annual variation in population growth rate was strongly negatively related to amount of precipitation during winter (i.e. high growth rates occurred when winters were dry). There was no significant relationship between the NAO-index and the population growth rate. However, there was an interaction between population density and the climatic variables, i.e. the effect of climate was stronger at high densities. These results support the view that population fluctuations of arctic ungulates are strongly influenced by stochastic variation in climate.     

Phylogeny of calcareous dinoflagellates as inferred from ITS and ribosomal sequence data

The phylogenetic relationships of calcareous dinoflagellates (i.e., Calciodinellaceae and Thoracosphaera) are investigated. Molecular data from the ribosomal 5.8S rRNA and highly conserved motifs of the ITS1 show Calciodinellaceae s.l. to be monophyletic when few non-calcareous taxa are included. They segregate into three monophyletic assemblages in a molecular analysis that considers the 5.8S rRNA and both the Internal Transcribed Spacer regions ITS1 and ITS2: a clade comprising species of Ensiculifera and Pentapharsodinium (E/P-clade), Scrippsiella s.l. (including fossil-based taxa such as Calciodinellum and Calcigonellum), and a heterogeneous group (T/P-clade) of calcareous (e.g., Thoracosphaera) and non-calcareous taxa (e.g., the highly toxic Pfiesteria). The potential to produce calcareous structures is considered as apomorphic within alveolates, and non-calcareous taxa nesting with calcareous dinoflagellates may have reduced calcification secondarily. Molecular results do not contradict general evolutionary scenarios provided by previous morphological (mainly paleontological) investigations.     

Increasing activity and thermal resistance of Bacillus gibsonii alkaline protease (BgAP) by directed evolution

Bacillus gibsonii Alkaline Protease (BgAP) is a recently reported subtilisin protease exhibiting activity and stability properties suitable for applications in laundry and dish washing detergents. However, BgAP suffers from a significant decrease of activity at low temperatures. In order to increase BgAP activity at 15°C, a directed evolution campaign based on the SeSaM random mutagenesis method was performed. An optimized microtiter plate expression system in B. subtilis was established and classical proteolytic detection methods were adapted for high throughput screening. In parallel, the libraries were screened for increased residual proteolytic activity after incubation at 58°C. Three iterative rounds of directed BgAP evolution yielded a set of BgAP variants with increased specific activity (K_cat) at 15°C and increased thermal resistance. Recombination of both sets of amino acid substitutions resulted finally in variant MF1 with a 1.5‐fold increased specific activity (15°C) and over 100 times prolonged half‐life at 60°C (224 min compared to 2 min of the WT BgAP). None of the introduced amino acid substitutions were close to the active site of BgAP. Activity‐altering amino acid substitutions were from non‐charged to non‐charged or from sterically demanding to less demanding. Thermal stability improvements were achieved by substitutions to negatively charged amino acids in loop areas of the BgAP surface which probably fostered ionic and hydrogen bonds interactions. Biotechnol. Bioeng. 2013; 110: 711–720. © 2012 Wiley Periodicals, Inc.     

Size-biased allocation of prey from male to offspring via female: family conflicts, prey selection, and evolution of sexual size dimorphism in raptors

In birds with bi-parental care, the provisioning link between prey capture and delivery to dependent offspring is regarded as often symmetric between the mates. However, in raptors, the larger female usually broods and feeds the nestlings, while the smaller male provides food for the family, assisted by the female in the latter part of the nestling period, if at all. Prey items are relatively large and often impossible for nestlings to handle without extended maternal assistance. We video-recorded prey delivery and handling in nests of a raptor with a wide diet, the Eurasian kestrel Falco tinnunculus, and simultaneously observed prey transfer from male to female outside the nest. The male selectively allocated larger items, in particular birds and larger mammals, to the female for further processing and feeding of nestlings, and smaller items, in particular lizards and smaller mammals, directly to the nestlings for unassisted feeding. Hence, from the video, the female appeared to have captured larger prey than the male, while in reality no difference existed. The female’s size-biased interception of the male’s prey provisioning line would maximize the male’s foraging time, and maximize the female’s control of the allocation of food between her own need and that of the offspring. The male would maximize his control of food allocation by capturing smaller prey. This conflict would select for larger dominant females and smaller energy-efficient males, and induce stronger selection the longer the female depends on the male for self-feeding, as a proportion of the offspring dependence period.     

Sensitivity and Robustness in Covalent Modification Cycles with a Bifunctional Converter Enzyme

Regulation by covalent modification is a common mechanism to transmit signals in biological systems. The modifying reactions are catalyzed either by two distinct converter enzymes or by a single bifunctional enzyme (which may employ either one or two catalytic sites for its opposing activities). The reason for this diversification is unclear, but contemporary theoretical models predict that systems with distinct converter enzymes can exhibit enhanced sensitivity to input signals whereas bifunctional enzymes with two catalytic sites are believed to generate robustness against variations in system’s components. However, experiments indicate that bifunctional enzymes can also exhibit enhanced sensitivity due to the zero-order effect, raising the question whether both phenomena could be understood within a common mechanistic model. Here, I argue that this is, indeed, the case. Specifically, I show that bifunctional enzymes with two catalytic sites can exhibit both ultrasensitivity and concentration robustness, depending on the kinetic operating regime of the enzyme’s opposing activities. The model predictions are discussed in the context of experimental observations of ultrasensitivity and concentration robustness in the uridylylation cycle of the PII protein, and in the phosphorylation cycle of the isocitrate dehydrogenase, respectively.     

The Quest for Comparability: Studying the Invariance of the Teachers’ Sense of Self-Efficacy (TSES) Measure across Countries

Teachers’ self-efficacy is an important motivational construct that is positively related to a variety of outcomes for both the teachers and their students. This study addresses challenges associated with the commonly used ‘Teachers’ Sense of Self-Efficacy (TSES)’ measure across countries and provides a synergism between substantive research on teachers’ self-efficacy and the novel methodological approach of exploratory structural equation modeling (ESEM). These challenges include adequately representing the conceptual overlap between the facets of self-efficacy in a measurement model (cross-loadings) and comparing means and factor structures across countries (measurement invariance). On the basis of the OECD Teaching and Learning International Survey (TALIS) 2013 data set comprising 32 countries (N = 164,687), we investigate the effects of cross-loadings in the TSES measurement model on the results of measurement invariance testing and the estimation of relations to external constructs (i.e., working experience, job satisfaction). To further test the robustness of our results, we replicate the 32-countries analyses for three selected sub-groups of countries (i.e., Nordic, East and South-East Asian, and Anglo-Saxon country clusters). For each of the TALIS 2013 participating countries, we found that the factor structure of the self-efficacy measure is better represented by ESEM than by confirmatory factor analysis (CFA) models that do not allow for cross-loadings. For both ESEM and CFA, only metric invariance could be achieved. Nevertheless, invariance levels beyond metric invariance are better achieved with ESEM within selected country clusters. Moreover, the existence of cross-loadings did not affect the relations between the dimensions of teachers’ self-efficacy and external constructs. Overall, this study shows that a conceptual overlap between the facets of self-efficacy exists and can be well-represented by ESEM. We further argue for the cross-cultural generalizability of the corresponding measurement model.     

Development of a novel expression,ZIMAX/KZI,for determination of the counter-anion effect on the antimicrobial activity of tetrabutylammonium salts

Due to the increasing number of strains of drug-resistant bacteria, the development of new antibiotics has become increasingly important. The antibacterial properties of quaternary amines and their derivatives on both Gram-positive and Gram-negative bacteria are well known. However, an encompassing study with specific emphasis on the role of the counter-anion has not been reported in the literature. By monitoring the Zone of Inhibition of various concentrations of tetrabutylammonium (TBA) salts, we observed that the counter anion plays a significant role in activity. We developed a novel method of reporting activity using zone of inhibition tests (ZI_MAX/K_ZI) and found it to be strongly correlated with the minimum inhibitory concentration (MIC).