Structural characterization of a new binding motif and a novel binding mode in group 2 WW domains

Formin homology 1 (FH1), is a long proline-rich region of formins, shown to bind to five WW containing proteins named formin binding proteins (FBPs). FH1 has several potential binding regions but only the PPLPx motif and its interaction with FBP11WW1 has been characterized structurally. To detect whether additional motifs exist in FH1, we synthesized five peptides and investigated their interaction with FBP28WW2, FBP11WW1 and FBP11WW2 domains. Peptides of sequence PTPPPLPP (positive control), PPPLIPPPP and PPLIPPPP (new motifs) interact with the domains with micromolar affinity. We observed that FBP28WW2 and FBP11WW2 behave differently from FBP11WW1 in terms of motif selection and affinity, since they prefer a doubly interrupted proline stretch of sequence PPLIPP. We determined the NMR structure of three complexes involving the FBP28WW2 domain and the three ligands. Depending on the peptide under study, the domain interacts with two proline residues accommodated in either the XP or the XP2 groove. This difference represents a one-turn displacement of the domain along the ligand sequence. To understand what drives this behavior, we performed further structural studies with the FBP11WW1 and a mutant of FBP28WW2 mimicking the XP2 groove of FBP11WW1. Our observations suggest that the nature of the XP2 groove and the balance of flexibility/rigidity around loop 1 of the domain contribute to the selection of the final ligand positioning in fully independent domains. Additionally, we analyzed the binding of a double WW domain region, FBP11WW1-2, to a long stretch of FH1 using fluorescence spectroscopy and NMR titrations. With this we show that the presence of two consecutive WW domains may also influence the selection of the binding mode, particularly if both domains can interact with consecutive motifs in the ligand. Our results represent the first observation of protein-ligand recognition where a pair of WW and two consecutive motifs in a ligand participate simultaneously.     

A putative gene cluster for aminoarabinose biosynthesis is essential for Burkholderia cenocepacia viability

Using a conditional mutagenesis strategy we demonstrate here that a gene cluster encoding putative aminoarabinose (Ara4N) biosynthesis enzymes is essential for the viability of Burkholderia cenocepacia. Loss of viability is associated with dramatic changes in bacterial cell morphology and ultrastructure, increased permeability to propidium iodide, and sensitivity to sodium dodecyl sulfate, suggesting a general cell envelope defect caused by the lack of Ara4N.     

Feeding location affects demographic performance of cabbage aphids on winter canola

The cabbage aphid, Brevicoryne brassicae L. (Hemiptera: Aphididae), is a perennial pest that specializes on plants of the Brassicaceae family, attacking winter canola (Brassica napus L.) mainly during and after flowering. Under field conditions, cabbage aphid colonizes the upper flowering canopy. Population dynamics of aphids in the flowering canopy could be regulated by differences in either plant quality (bottom‐up) or predatory (top‐down) forces. The goal of our study was to determine the effect of feeding location on cabbage aphid demography. A stage‐structured matrix population model was constructed for aphids restricted to reproductive or vegetative plant tissues of canola. We found that feeding location had a large impact on demography of cabbage aphid; the finite rate of increase (λ ± SEM) was higher when aphids were restricted to reproductive tissues, compared to aphids feeding on vegetative tissues: 1.25 ± 0.01 vs. 1.17 ± 0.01 (leaves). Aphids confined to reproductive tissues with higher λ exhibited shorter generation times (T = 14.2 ± 0.2 days) and 53–75% higher net reproductive rates (R₀ = 23.3 ± 1.7) than aphids feeding on vegetative tissues. Prospective analyses showed that there was a nymph‐skewed stable stage distribution, and elasticity values revealed that λ is most sensitive to changes in stasis of adults staying in the adult stage and to adult survival. Retrospective analyses indicated that variation in adult fecundity (value of 0.05) had the largest effect on population dynamics but collectively, growth of nymphal stage 2–3, 3–4, and 4 to adult accounted for most of the difference in λ between the treatments. Monitoring programs should target adults and penultimate instars colonizing reproductive tissues of canola plants in the field as aphids on these plant structures contribute most to population growth.     

Does Ownership Matter? An Overview of Systematic Reviews of the Performance of Private For-Profit,Private Not-For-Profit and Public Healthcare Providers

Introduction

Ownership of healthcare providers has been considered as one factor that might influence their health and healthcare related performance. The aim of this article was to provide an overview of what is known about the effects on economic, administrative and health related outcomes of different types of ownership of healthcare providers -namely public, private non-for-profit (PNFP) and private for-profit (PFP)- based on the findings of systematic reviews (SR).

Methods and Findings

An overview of systematic reviews was performed. Different databases were searched in order to select SRs according to an explicit comprehensive criterion. Included SRs were assessed to determine their methodological quality. Of the 5918 references reviewed, fifteen SR were included, but six of them were rated as having major limitations, so they weren''t incorporated in the analyses. According to the nine analyzed SR, ownership does seem to have an effect on health and healthcare related outcomes. In the comparison of PFP and PNFP providers, significant differences in terms of mortality of patients and payments to facilities have been found, both being higher in PFP facilities. In terms of quality and economic indicators such as efficiency, there are no concluding results. When comparing PNFP and public providers, as well as for PFP and public providers, no clear differences were found.

Conclusion

PFP providers seem to have worst results than their PNFP counterparts, but there are still important evidence gaps in the literature that needs to be covered, including the comparison between public and both PFP and PNFP providers. More research is needed in low and middle income countries to understand the impact on and development of healthcare delivery systems.     

Over-expression of potato virus X TGBp1 movement protein in transgenic tobacco plants causes developmental and metabolic alterations.

Transgenic Nicotiana tabacum plants expressing the TGBp1 movement protein of potato virus X (PVX) were studied to investigate the effects caused by this protein on plant physiology and development. TGBp1 caused consistent reductions of size and weight in different organs of these plants; however shoot-to-root ratios were similar to those of control plants. Transgenic seedlings showed smaller root meristems and calli derived from TGBp1 leaves grew at a slower rate through successive subcultures. Microscopic observations of TGBp1 plants revealed flattened chloroplasts containing plastoglobuli-like bodies. Further analyses showed a considerable reduction in photosynthetic rate, lower starch levels in leaves and roots, higher nitrate accumulation in leaves and induction of pathogenesis-related (PR) protein genes. Since these changes were not observed when other PVX sequences were expressed in tobacco, we postulate that TGBp1 is an important symptom contributor in PVX infections.     

Syndecan-2 is a novel target of insulin-like growth factor binding protein-3 and is over-expressed in fibrosis

Extracellular matrix deposition and tissue scarring characterize the process of fibrosis. Transforming growth factor beta (TGFβ) and Insulin-like growth factor binding protein-3 (IGFBP-3) have been implicated in the pathogenesis of fibrosis in various tissues by inducing mesenchymal cell proliferation and extracellular matrix deposition. We identified Syndecan-2 (SDC2) as a gene induced by TGFβ in an IGFBP-3-dependent manner. TGFβ induction of SDC2 mRNA and protein required IGFBP-3. IGFBP-3 independently induced production of SDC2 in primary fibroblasts. Using an ex-vivo model of human skin in organ culture expressing IGFBP-3, we demonstrate that IGFBP-3 induces SDC2 ex vivo in human tissue. We also identified Mitogen-activated protein kinase-interacting kinase (Mknk2) as a gene induced by IGFBP-3. IGFBP-3 triggered Mknk2 phosphorylation resulting in its activation. Mknk2 independently induced SDC2 in human skin. Since IGFBP-3 is over-expressed in fibrotic tissues, we examined SDC2 levels in skin and lung tissues of patients with systemic sclerosis (SSc) and lung tissues of patients with idiopathic pulmonary fibrosis (IPF). SDC2 levels were increased in fibrotic dermal and lung tissues of patients with SSc and in lung tissues of patients with IPF. This is the first report describing elevated levels of SDC2 in fibrosis. Increased SDC2 expression is due, at least in part, to the activity of two pro-fibrotic factors, TGFβ and IGFBP-3.