NMR assignments of the GacS histidine-kinase periplasmic detection domain from <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> PAO1

Pseudomonas aeruginosa is a highly adaptable opportunistic pathogen. It can infect vulnerable patients such as those with cystic fibrosis or hospitalized in intensive care units where it is responsible for both acute and chronic infection. The switch between these infections is controlled by a complex regulatory system involving the central GacS/GacA two-component system that activates the production of two small non-coding RNAs. GacS is a histidine kinase harboring one periplasmic detection domain, two inner-membrane helices and three H1/D1/H2 cytoplasmic domains. By detecting a yet unknown signal, the GacS histidine-kinase periplasmic detection domain (GacSp) is predicted to play a key role in activating the GacS/GacA pathway. Here, we present the chemical shift assignment of 96 % of backbone atoms (HN, N, C, Cα, Cβ and Hα), 88 % aliphatic hydrogen atoms and 90 % of aliphatic carbon atoms of this domain. The NMR-chemical shift data, on the basis of Talos server secondary structure predictions, reveal that GacSp consists of 3 β-strands, 3 α-helices and a major loop devoid of secondary structures.     

Calcium Currents Are Enhanced by ��2��-1 Lacking Its Membrane Anchor

The accessory α₂δ subunits of voltage-gated calcium channels are membrane-anchored proteins, which are highly glycosylated, possess multiple disulfide bonds, and are post-translationally cleaved into α₂ and δ. All α₂δ subunits have a C-terminal hydrophobic, potentially trans-membrane domain and were described as type I transmembrane proteins, but we found evidence that they can be glycosylphosphatidylinositol-anchored. To probe further the function of membrane anchoring in α₂δ subunits, we have now examined the properties of α₂δ-1 constructs truncated at their putative glycosylphosphatidylinositol anchor site, located before the C-terminal hydrophobic domain (α₂δ-1ΔC-term). We find that the majority of α₂δ-1ΔC-term is soluble and secreted into the medium, but unexpectedly, some of the protein remains associated with detergent-resistant membranes, also termed lipid rafts, and is extrinsically bound to the plasma membrane. Furthermore, heterologous co-expression of α₂δ-1ΔC-term with Ca_V2.1/β1b results in a substantial enhancement of the calcium channel currents, albeit less than that produced by wild-type α₂δ-1. These results call into question the role of membrane anchoring of α₂δ subunits for calcium current enhancement.     

Differential requirements for the CENP-O complex reveal parallel PLK1 kinetochore recruitment pathways

Similar to other core biological processes, the vast majority of cell division components are essential for viability across human cell lines. However, recent genome-wide screens have identified a number of proteins that exhibit cell line–specific essentiality. Defining the behaviors of these proteins is critical to our understanding of complex biological processes. Here, we harness differential essentiality to reveal the contributions of the four-subunit centromere-localized CENP-O complex, whose precise function has been difficult to define. Our results support a model in which the CENP-O complex and BUB1 act in parallel pathways to recruit a threshold level of PLK1 to mitotic kinetochores, ensuring accurate chromosome segregation. We demonstrate that targeted changes to either pathway sensitizes cells to the loss of the other component, resulting in cell-state dependent requirements. This approach also highlights the advantage of comparing phenotypes across diverse cell lines to define critical functional contributions and behaviors that could be exploited for the targeted treatment of disease.     

Protein Topology Prediction Algorithms Systematically Investigated in the Yeast Saccharomyces cerevisiae

Membrane proteins perform a variety of functions, all crucially dependent on their orientation in the membrane. However, neither the exact number of transmembrane domains (TMDs) nor the topology of most proteins have been experimentally determined. Due to this, most scientists rely primarily on prediction algorithms to determine topology and TMD assignments. Since these can give contradictory results, single‐algorithm‐based predictions are unreliable. To map the extent of potential misanalysis, the predictions of nine algorithms on the yeast proteome are compared and it is found that they have little agreement when predicting TMD number and termini orientation. To view all predictions in parallel, a webpage called TopologYeast: http://www.weizmann.ac.il/molgen/TopologYeast was created. Each algorithm is compared with experimental data and a poor agreement is found. The analysis suggests that more systematic data on protein topology are required to increase the training sets for prediction algorithms and to have accurate knowledge of membrane protein topology.     

Self-perceived oral health and access to care in diabetic and non-diabetic women: A qualitative study

BackgroundAccess to dental care is of growing importance to individuals with chronic illnesses and policymakers alike.ObjectivesTo explore personal views of diabetic and non-diabetic women regarding their oral health and access to dental care.MethodsA qualitative study was performed involving 6 diabetic and 6 non-diabetic female hospital attendees. A thematic framework approach was used to analyze the interviews.ResultsResponses yield three emerging central themes: oral health impact, self-maintained oral health and dental service delivery and costs. Personal views varied particularly in oral health self-perception and visiting the dentist. Certain drawbacks shadowed the quality of care in governmental and private sectors. Response variations were observed among diabetics and non-diabetics.ConclusionWithin the study limits, female hospital attendees had variable responses on how they looked upon their oral health. A variation in oral hygiene practices and visiting the dentist was observed. Participants believed that governmental and private clinics offered high quality care, with high treatment costs in private clinics and long waiting times in the public service being the main drawbacks. Practical Significance: Importance of supporting patients, educating practitioners and alarming policymakers about the impact of oral health problems and access to dental care.     

Radiation effects on the structure property of CA 80 for measuring personnel radiation doses

The effects of fast and albedo neutrons and alpha particles on the molecular fractions of cellulose nitrate type CA 80, as a tissue equivalent material have been measured through the use of gel permeation chromatography. The samples in the form of thin sheets were exposed to fission neutron fluences in the range 10⁵–10¹⁰ n/cm² and to different energies of alpha particles in the range 1–5 MeV. The effects of irradiation on paraffin phantom, application of lithium borate radiator to the sample and storage of irradiated specimens for different periods were measured. The results indicated that CA80 eluted in one main fraction. Irradiation with neutrons or alpha particles caused fragmentation of this fraction to smaller molecules. Small and acceptable fade of the induced changes by radiation in the polymer due to storage for more than one month were noticed. The possibility of applying this technique in radiation dosimetry was discussed.