Molecular organization of cytokinesis node predicts the constriction rate of the contractile ring

The molecular organization of cytokinesis proteins governs contractile ring function. We used single molecule localization microscopy in live cells to elucidate the molecular organization of cytokinesis proteins and relate it to the constriction rate of the contractile ring. Wild-type fission yeast cells assemble contractile rings by the coalescence of cortical proteins complexes called nodes whereas cells without Anillin/Mid1p (Δmid1) lack visible nodes yet assemble contractile rings competent for constriction from the looping of strands. We leveraged the Δmid1 contractile ring assembly mechanism to determine how two distinct molecular organizations, nodes versus strands, can yield functional contractile rings. Contrary to previous interpretations, nodes assemble in Δmid1 cells. Our results suggest that Myo2p heads condense upon interaction with actin filaments and an excess number of Myo2p heads bound to actin filaments hinders constriction thus reducing the constriction rate. Our work establishes a predictive correlation between the molecular organization of nodes and the behavior of the contractile ring.     

Ecosystem Service Provision by Secondary Forests in Shifting Cultivation Areas Remains Poorly Understood

Human Ecology - It is often asserted that secondary forests (SF) provide inferior forest-based ecosystem services (ES), but there is limited research to generalize this claim. Here, we review...     

Rosetta gen. nov. (Chlorophyta): Resolving the identity of red snow algal rosettes

Thick-walled rosette-like snow algae were long thought to be a life stage of various other species of snow algae. Rosette-like cells have not been cultured, but by manually isolating cells from 38 field samples in southern British Columbia, we assigned a variety of rosette morphologies to DNA sequence. Phylogenetic analysis of Rubisco large-subunit (rbcL) gene, ribosomal internal transcribed spacer 2 (ITS2) rRNA region, and 18S rRNA gene revealed that the rosette-like cells form a new clade within the phylogroup Chloromonadinia. Based on these data, we designate a new genus, Rosetta, which comprises five novel species: R. castellata, R. floranivea, R. stellaria, R. rubriterra, and R. papavera. In a survey of 762 snow samples from British Columbia, we observed R. floranivea exclusively on snow overlying high-elevation glaciers, whereas R. castellata was observed at lower elevations, near the tree line. The other three species were rarely observed. Spherical red cells enveloped in a thin translucent sac were conspecific with Rosetta, possibly a developmental stage. These results highlight the unexplored diversity among snow algae and emphasize the utility of single-cell isolation to advance the centuries-old problem of disentangling life stages and cryptic species.     

Behavioral strategies for the reduction of pain and anxiety associated with orthopedic trauma

This study examined the efficacy of behavioral strategies in alleviating pain and anxiety associated with severe orthopedic trauma. Sixty-four patients with multiple fractures were divided into four groups: (1) control, (2) attention only, (3) EMG biofeedback-assisted relaxation, and (4) audiotaped relaxation training. All were measured over at least six sessions, or as long as hospital stay permitted. Significant between group differences were found on the following: systolic blood pressure, peripheral temperature, subjective units of discomfort, state anxiety, with a trend for use of sleep medications. No differences were found on other vital signs, EMG recordings, or other medications. EMG-biofeedback relaxation and relaxation training were relatively equivalent for all measures, and little or no change was observed for those patients who received attention only or served as controls.     

Life history characteristics of Orius insidiosus (Say) fed diets of soybean aphid, Aphis glycines Matsumura and soybean thrips, Neohydatothrips variabilis (Beach)

Field studies in soybeans have demonstrated that the endemic predator, Orius insidiosus (Say), is an important natural enemy of the soybean aphid, Aphis glycines Matsumura. Soybean thrips, Neohydatothrips variabilis (Beach), serve as an important prey resource for O. insidiosus in soybeans and may be important in sustaining O. insidiosus populations before the arrival of soybean aphid. Because soybean aphid is new to the US soybean system, the effects of a mixed diet of soybean aphid and soybean thrips on O. insidiosus life history is not known. We measured the survival, development, and reproduction of O. insidiosus when fed soybean thrips, and a mixed prey diet of soybean aphids and soybean thrips, and compared these results to a previous study of O. insidiosus life history fed soybean aphid alone. Nymphal development to adulthood (15.9 days) and fecundity (68.8 eggs per female) was improved for O. insidiosus fed ad libitum soybean thrips daily compared to O. insidiosus fed ad libitum soybean aphids daily. The contribution of alternative prey to O. insidiosus life history characteristics can be complex depending on the amount and quality of a particular prey item. At low levels of prey, the addition of prey appears to enhance O. insidiosus survival, development, and fecundity. However, as predators are fed more often, the predator’s response depends on the type of prey that predominates in the mixed prey diet. We discuss soybean thrips impact on O. insidiosus population ecology and soybean aphid dynamics.     

Enhanced transduction of dendritic cells by FcgammaRI-targeted adenovirus vectors

BACKGROUND: The high affinity Fcgamma receptor I (FcgammaRI; aka CD64) is expressed by dendritic cells (DC) and antigens targeted to this receptor elicit enhanced immune responses. This study was designed to test the hypothesis that targeting an adenoviral (Ad) vector to FcgammaRI would lead to enhanced transduction of DC and an improved immune response to vector-encoded antigens. METHODS: A bispecific adaptor molecule consisting of a trimeric adenovirus fiber-binding moiety fused to a single-chain antibody specific for human FcgammaRI was generated. Transduction of cultured cells, including human DC, by the FcgammaRI-targeted Ad was then evaluated using reporter genes (GFP, luciferase). Immunophenotypic and functional characteristics of vector-transduced DC were also measured by flow cytometry, cytokine ELISA and mixed lymphocyte reaction (MLR); antigen-specific stimulation of autologous CD8(+) T cells was evaluated using vectors encoding cytomegalovirus (CMV) pp65. RESULTS: FcgammaRI-targeted Ad transduced primary DC with 10-15-fold greater efficiency than unmodified Ad or Ad vectors complexed to an adaptor protein that targeted an irrelevant receptor. However, FcgammaRI-targeting had no effect of Ad-induced activation of DC, as measured by cytokine release or expression of cell surface activation markers. Finally, FcgammaRI-targeting of vectors encoding CMV pp65 resulted in an increase in the activation of antigen-specific autologous human CD8(+) T cells. CONCLUSIONS: FcgammaRI-targeting significantly enhances the efficiency of Ad vector-mediated gene transfer in primary human DC, and results in an improved immune response to a vector-encoded antigen.     

Discovering and detecting transposable elements in genome sequences

The contribution of transposable elements (TEs) to genome structure and evolution as well as their impact on genome sequencing, assembly, annotation and alignment has generated increasing interest in developing new methods for their computational analysis. Here we review the diversity of innovative approaches to identify and annotate TEs in the post-genomic era, covering both the discovery of new TE families and the detection of individual TE copies in genome sequences. These approaches span a broad spectrum in computational biology including de novo, homology-based, structure-based and comparative genomic methods. We conclude that the integration and visualization of multiple approaches and the development of new conceptual representations for TE annotation will further advance the computational analysis of this dynamic component of the genome.     

Subcellular localisation of <Emphasis Type="Italic">Medicago truncatula</Emphasis>9/13-hydroperoxide lyase reveals a new localisation pattern and activation mechanism for CYP74C enzymes

Background  

Hydroperoxide lyase (HPL) is a key enzyme in plant oxylipin metabolism that catalyses the cleavage of polyunsaturated fatty acid hydroperoxides produced by the action of lipoxygenase (LOX) to volatile aldehydes and oxo acids. The synthesis of these volatile aldehydes is rapidly induced in plant tissues upon mechanical wounding and insect or pathogen attack. Together with their direct defence role towards different pathogens, these compounds are believed to play an important role in signalling within and between plants, and in the molecular cross-talk between plants and other organisms surrounding them. We have recently described the targeting of a seed 9-HPL to microsomes and putative lipid bodies and were interested to compare the localisation patterns of both a 13-HPL and a 9/13-HPL from Medicago truncatula, which were known to be expressed in leaves and roots, respectively.     

Inactivation of the DnaB helicase leads to the collapse and degradation of the replication fork: a comparison to UV-induced arrest

Replication forks face a variety of structurally diverse impediments that can prevent them from completing their task. The mechanism by which cells overcome these hurdles is likely to vary depending on the nature of the obstacle and the strand in which the impediment is encountered. Both UV-induced DNA damage and thermosensitive replication proteins have been used in model systems to inhibit DNA replication and characterize the mechanism by which it recovers. In this study, we examined the molecular events that occur at replication forks following inactivation of a thermosensitive DnaB helicase and found that they are distinct from those that occur following arrest at UV-induced DNA damage. Following UV-induced DNA damage, the integrity of replication forks is maintained and protected from extensive degradation by RecA, RecF, RecO, and RecR until replication can resume. By contrast, inactivation of DnaB results in extensive degradation of the nascent and leading-strand template DNA and a loss of replication fork integrity as monitored by two-dimensional agarose gel analysis. The degradation that occurs following DnaB inactivation partially depends on several genes, including recF, recO, recR, recJ, recG, and xonA. Furthermore, the thermosensitive DnaB allele prevents UV-induced DNA degradation from occurring following arrest even at the permissive temperature, suggesting a role for DnaB prior to loading of the RecFOR proteins. We discuss these observations in relation to potential models for both UV-induced and DnaB(Ts)-mediated replication inhibition.