Tyrosine 331 and phenylalanine 334 in Clostridium perfringens alpha-toxin are essential for cytotoxic activity

Differences in the biological properties of the Clostridium perfringens phospholipase C (alpha-toxin) and the C. bifermentans phospholipase C (Cbp) have been attributed to differences in their carboxy-terminal domains. Three residues in the carboxy-terminal domain of alpha-toxin, which have been proposed to play a role in membrane recognition (D269, Y331 and F334), are not conserved in Cbp (Y, L and I respectively). We have characterised D269Y, Y331L and F334I variant forms of alpha-toxin. Variant D269Y had reduced phospholipase C activity towards aggregated egg yolk phospholipid but increased haemolytic and cytotoxic activity. Variants Y331L and F334I showed a reduction in phospholipase C, haemolytic and cytotoxic activities indicating that these substitutions contribute to the reduced haemolytic and cytotoxic activity of Cbp.     

A selective insecticidal protein from Pseudomonas mosselii for corn rootworm control

The coleopteran insect western corn rootworm (WCR, Diabrotica virgifera virgifera) is an economically important pest in North America and Europe. Transgenic corn plants producing Bacillus thuringiensis (Bt) insecticidal proteins have been useful against this devastating pest, but evolution of resistance has reduced their efficacy. Here, we report the discovery of a novel insecticidal protein, PIP‐47Aa, from an isolate of Pseudomonas mosselii. PIP‐47Aa sequence shows no shared motifs, domains or signatures with other known proteins. Recombinant PIP‐47Aa kills WCR, two other corn rootworm pests (Diabrotica barberi and Diabrotica undecimpunctata howardi) and two other beetle species (Diabrotica speciosa and Phyllotreta cruciferae), but it was not toxic to the spotted lady beetle (Coleomegilla maculata) or seven species of Lepidoptera and Hemiptera. Transgenic corn plants expressing PIP‐47Aa show significant protection from root damage by WCR. PIP‐47Aa kills a WCR strain resistant to mCry3A and does not share rootworm midgut binding sites with mCry3A or AfIP‐1A/1B from Alcaligenes that acts like Cry34Ab1/Cry35Ab1. Our results indicate that PIP‐47Aa is a novel insecticidal protein for controlling the corn rootworm pests.     

How Accurate are Recalls of Self‐Weighing Frequency? Data from a 24‐Month Randomized Trial

Introduction

Self‐weighing is an important component of self‐monitoring during weight loss. However, methods of measuring self‐weighing frequency need to be validated. This analysis compared self‐reported and objective weighing frequency.

Methods

Data came from a 24‐month randomized controlled trial. Participants received 12 months of a behavioral weight‐loss program and were randomly assigned to (1) daily self‐weighing, (2) weekly weighing, or (3) no weighing (excluded from analysis). Objective weighing frequency was measured by Wi‐Fi enabled scales, and self‐reported weighing frequency was assessed every 6 months by questionnaire. Objective weights were categorized to match the scale of the self‐report measure.

Results

At 12 months, there was 80.8% agreement between self‐reported and objective weighing frequency (weighted kappa = 0.67; P < 0.001). At 24 months, agreement decreased to 48.5% (kappa = 0.27; P < 0.001). At both time points in which disagreements occurred, self‐reported frequencies were generally greater than objectively assessed weighing. Both self‐reported and objectively assessed weighing frequency was associated with weight loss at 12 and 24 months (P < 0.001).

Conclusions

Self‐reported weighing frequency is modestly correlated with objective weighing frequency; however, both are associated with weight change over time. Objective assessment of weighing frequency should be used to avoid overestimating actual frequency.

     

The Aer2 receptor from Vibrio cholerae is a dual PAS‐heme oxygen sensor

The diarrheal pathogen Vibrio cholerae navigates complex environments using three chemosensory systems and 44–45 chemoreceptors. Chemosensory cluster II modulates chemotaxis, whereas clusters I and III have unknown functions. Ligands have been identified for only five V. cholerae chemoreceptors. Here, we report that the cluster III receptor, VcAer2, binds and responds to O₂. VcAer2 is an ortholog of Pseudomonas aeruginosa Aer2 (PaAer2) but differs in that VcAer2 has two, rather than one, N‐terminal PAS domain. We have determined that both PAS1 and PAS2 form homodimers and bind penta‐coordinate b‐type heme via an Eη‐His residue. Heme binding to PAS1 required the entire PAS core, but receptor function also required the N‐terminal cap. PAS2 functioned as an O₂‐sensor [ , 19 μM], utilizing the same Iβ Trp (W276) as PaAer2 to stabilize O₂. The crystal structure of PAS2‐W276L was similar to that of PaAer2‐PAS but resided in an active conformation mimicking the ligand‐bound state, consistent with its signal‐on phenotype. PAS1 also bound O₂ [ , 12 μM], although O₂ binding was stabilized by either a Trp residue or Tyr residue. Moreover, PAS1 appeared to function as a signal modulator, regulating O₂‐mediated signaling from PAS2 and resulting in activation of the cluster III chemosensory pathway.     

Pollen Organs and Seeds with Eucommiidites Pollen

The characteristic Mesozoic pollen genus Eucommiidites is described from pollen organs and seeds recovered in Cretaceous strata of North America and Europe. The pollen organs are from the lowermost Upper Cretaceous (Cenomanian) of Texas and are referred to Erdtmanitheca texensis gen. et sp. nov. They are spherical heads, composed of numerous, densely crowded, radiating pollen sacs that contain abundant well-preserved pollen. Combined LM, SEM and TEM investigations show that the pollen grains each have a distinct distal colpus flanked by two lateral colpi in an equatorial position. Pollen wall ultrastructure is gymnospermous with a thick lamellate inner layer (endexine) and an outer layer (ektexine) composed of a granular inner part and a homogeneous outer part. The endexine is thickened in the region of the colpi. Small seeds from the Lower Cretaceous (upper Berriasian to Valanginian) of Bornholm, Denmark contain abundant Eucommiidites pollen in their micropyles. The seeds are referred to Erdtmanispermum balticum gen. et sp. nov. They are ovoid, and weakly triangular in transverse section and gradually taper at the apex into an elongated tube. The megaspore membrane is granular and well developed, and apparently surrounded by three separate tissues interpreted as nucellus, a thin inner integument and a sclerified outer envelope. Eucommiidites pollen in the micropyles of the seeds has a laminated endexine and an ektexine comprising two homogeneous parts separated by a granular layer. Réévaluation of other seeds known to contain Eucommiidites pollen indicates that they share significant similarities with Erdlmanispermum and that they may have been produced by closely related plants. Comparison of “Eucommiidites plants” with other seed plants suggests that they are probably most closely related to the anthophytes comprising Bennettitales, Pentoxylales, Gnetales and angiosperms.     

Upatoia barnardii gen. et sp. nov., an araucarian pollen cone with in situ pollen from the Late Cretaceous (Santonian) of Georgia,USA

Upatoia barnardii gen. et sp. nov., a conifer pollen cone from the Late Cretaceous (Santonian) Eutaw Formation of Upatoi Creek, Georgia, USA, is known from lignified and fusainised mesofossils that preserve its three-dimensional structure. The cone consists of numerous helically arranged microsporophylls, each composed of a thin stalk and distal lamina. Three elongate pollen sacs are attached to the base of the lamina. Pollen grains isolated from the pollen sacs are relatively large (52 – 75 μm), spheroidal to ellipsoidal in outline, lack sacci, and have a thickened equatorial exine that is often strongly folded. Pollen of Upatoia barnardii indicates a close relationship to extant Araucariaceae. Microsporophylls of U. barnardii confirm suggestions from previous studies of fossil material that some Mesozoic Araucariaceae had only three pollen sacs per microsporophyll, in contrast to extant species that often have more than ten pollen sacs per microsporophyll.     

Explaining the distribution of breeding and dispersal syndromes in conifers

The evolution of plants exhibiting different sexes, or dioecy, is correlated with a number of ecological and life-history traits such as woody growth form and animal-dispersed seeds, but the underlying causes of these associations are unclear. Previous work in seed plants has suggested that the evolution of fleshy cones or seeds may favour dioecy. In this study, we use a well-sampled molecular phylogeny of conifers to show that although dioecy and fleshiness strongly co-occur at the species level, this relationship has not resulted from numerous separate origins of this trait combination or from differential rates of diversification. Instead, we suggest that two character combinations—the ancestral dry-monoecious condition and the derived fleshy-dioecious condition—have persisted in conifers longer than other combinations over evolutionary time. The persistence of these trait combinations appears to reflect differences in the rate of successful transition into and out of these character states over time, as well as the geographical restriction of species with rare combinations and their consequent vulnerability to extinction. In general, we argue that such persistence explanations should be considered alongside ‘key innovation’ hypotheses in explaining the phylogenetic distribution of traits.     

Alzheimer’s Disease: Analyzing the Missing Heritability

Alzheimer’s disease (AD) is a complex disorder influenced by environmental and genetic factors. Recent work has identified 11 AD markers in 10 loci. We used Genome-wide Complex Trait Analysis to analyze >2 million SNPs for 10,922 individuals from the Alzheimer’s Disease Genetics Consortium to assess the phenotypic variance explained first by known late-onset AD loci, and then by all SNPs in the Alzheimer’s Disease Genetics Consortium dataset. In all, 33% of total phenotypic variance is explained by all common SNPs. APOE alone explained 6% and other known markers 2%, meaning more than 25% of phenotypic variance remains unexplained by known markers, but is tagged by common SNPs included on genotyping arrays or imputed with HapMap genotypes. Novel AD markers that explain large amounts of phenotypic variance are likely to be rare and unidentifiable using genome-wide association studies. Based on our findings and the current direction of human genetics research, we suggest specific study designs for future studies to identify the remaining heritability of Alzheimer’s disease.