Do Small Headgroups of Phosphatidylethanolamine and Phosphatidic Acid Lead to a Similar Folding Pattern of the K+ Channel?

Phospholipid headgroups act as major determinants in proper folding of oligomeric membrane proteins. The K+-channel KcsA is the most popular model protein among these complexes. The presence of zwitterionic nonbilayer lipid phosphatidylethanolamine (PE) is crucial for efficient tetramerization and stabilization of KcsA in a lipid bilayer. In this study, the influence of PE on KcsA folding properties was analyzed by tryptophan fluorescence and acrylamide quenching experiments and compared with the effect of anionic phosphatidic acid (PA). The preliminary studies suggest that the small size and hydrogen bonding capability of the PE headgroup influences KcsA folding via a mechanism quite similar to that observed for anionic PA

Determination of nitrogen dioxide, sulfur dioxide, ozone, and ammonia in ambient air using the passive sampling method associated with ion chromatographic and potentiometric analyses

Concentrations of nitrogen dioxide (NO2), sulfur dioxide (SO2), ozone (O3), and ammonia (NH3) were determined in the ambient air of Al-Ain city over a year using the passive sampling method associated with ion chromatographic and potentiometric detections. IVL samplers were used for collecting nitrogen and sulfur dioxides whereas Ogawa samplers were used for collecting ozone and ammonia. Five sites representing the industrial, traffic, commercial, residential, and background regions of the city were monitored in the course of this investigation. Year average concentrations of ≤59.26, 15.15, 17.03, and 11.88 μg/m3 were obtained for NO2, SO2, O3, and NH3, respectively. These values are lower than the maxima recommended for ambient air quality standards by the local environmental agency and the world health organization. Results obtained were correlated with the three meteorological parameters: humidity, wind speed, and temperature recorded during the same period of time using the paired t test, probability p values, and correlation coefficients. Humidity and wind speed showed insignificant effects on NO2, SO2, O3, and NH3 concentrations at 95% confidence level. Temperature showed insignificant effects on the concentrations of NO2 and NH3 while significant effects on SO2 and O3 were observed. Nonlinear correlations (R2 ≤ 0.722) were obtained for the changes in measured concentrations with changes in the three meteorological parameters. Passive samplers were shown to be not only precise (RSD ≤ 13.57) but also of low cost, low technical demand, and expediency in monitoring different locations

Orientation and dynamics of transmembrane peptides: the power of simple models

In this review we discuss recent insights obtained from well-characterized model systems into the factors that determine the orientation and tilt angles of transmembrane peptides in lipid bilayers. We will compare tilt angles of synthetic peptides with those of natural peptides and proteins, and we will discuss how tilt can be modulated by hydrophobic mismatch between the thickness of the bilayer and the length of the membrane spanning part of the peptide or protein. In particular, we will focus on results obtained on tryptophan-flanked model peptides (WALP peptides) as a case study to illustrate possible consequences of hydrophobic mismatch in molecular detail and to highlight the importance of peptide dynamics for the experimental determination of tilt angles. We will conclude with discussing some future prospects and challenges concerning the use of simple peptide/lipid model systems as a tool to understand membrane structure and function

Non-Japanese Residents and the Earthquake : Reflections on our work thus far (Research Group on Non-Japanese Residents and the Earthquake)

BACKGROUND: Renal tubular epithelial cells (TECs) are one of the main targets of inflammatory insults during interstitial nephritis and kidney transplant rejection. While Th1 cells are know to be essential in the pathogenesis of rejection, the role of Th17 is still under debate. We hypothesize that TECs modulate the outcome of rejection process by production of distinct chemokines and cytokines that determine the attraction of different T-cell subsets. Therefore, we studied differential effects of activated human renal epithelial cells on T-cell migration. METHODS: Human primary TECs were stimulated by IFN-γ and TNF-α in vitro. Chemokines and cytokines produced by activated TECs were measured using Luminex or ELISA. Chemotaxis assay was performed using activated peripheral blood mononuclear cells composed of CD4+CXCR3+ and CD4+CCR6+ T cells migrating towards stimulated and unstimulated TECs. RESULTS: While activated TECs secreted abundant amounts of the pro-inflammatory cytokines IL-6 and IL-8, the T helper cell differentiation cytokines IL-1β, IL-12p70, IL-23 or TGF-β1 were not produced. The production of Th1 chemokines CXCL9, CXCL10 and CCL5 were significantly upregulated after TEC stimulation. In contrast, Th17 chemokine CCL20 could not be detected. Finally, activated TECs attracted significantly higher numbers of CD4+CXCR3+ T cells as compared to unstimulated TECs. No migration of CD4+CCR6+ T cells could be observed. CONCLUSION: Activated primary renal tubular epithelial cells do not attract Th17 cells nor produce cytokines promoting Th17 cell differentiation in our experimental system mimicking the proinflammatory microenvironment of rejection

Combgap contributes to recruitment of Polycomb group proteins in Drosophila

Polycomb group (PcG) proteins are responsible for maintaining the silenced transcriptional state of many developmentally regulated genes. PcG proteins are organized into multiprotein complexes that are recruited to DNA via cis-acting elements known as "Polycomb response elements" (PREs). In Drosophila, PREs consist of binding sites for many different DNA-binding proteins, some known and others unknown. Identification of these DNA-binding proteins is crucial to understanding the mechanism of PcG recruitment to PREs. We report here the identification of Combgap (Cg), a sequence-specific DNA-binding protein that is involved in recruitment of PcG proteins. Cg can bind directly to PREs via GTGT motifs and colocalizes with the PcG proteins Pleiohomeotic (Pho) and Polyhomeotic (Ph) at the majority of PREs in the genome. In addition, Cg colocalizes with Ph at a number of targets independent of Pho. Loss of Cg leads to decreased recruitment of Ph at only a subset of sites; some of these sites are binding sites for other Polycomb repressive complex 1 (PRC1) components, others are not. Our data suggest that Cg can recruit Ph in the absence of PRC1 and illustrate the diversity and redundancy of PcG protein recruitment mechanisms

Physiological and behavioural responses of broilers to controlled atmosphere stunning: implications for welfare

AbstractControlled atmosphere (gas) stunning (CAS) has the potential to improve the welfare of poultry at slaughter but there is a lack of consensus about which gas mixtures are most humane. The aim of this study was to evaluate the welfare consequences of different gas stunning approaches. Individual broilers were exposed to gas mixtures capable of inducing unconsciousness and euthanasia while their behavioural, cardiac, respiratory and neurophysiological responses were measured simultaneously. The approaches investigated included anoxia (N2or Ar with &lt; 2% residual O2), hypercapnic anoxia (30% CO2in Ar, 40% CO2in N2) and a biphasic method (40% CO2, 30% O2, 30% N2for 60 s followed by 80% CO2in air). Evaluation of the welfare implications of each approach centred on the likelihood of them inducing negative states or experiences during the conscious phase. Hypercapnic mixtures were associated with strong respiratory responses, while anoxic mixtures induced vigorous wing flapping. Electroencephalogram analysis using the correlation dimension (a non-linear measure of complexity) suggested that anoxic wing flapping occurred during periods in which a form of consciousness could not be excluded. Hypercapnic hyperoxygenation (biphasic approach) exacerbated respiratory responses but eliminated the possibility of vigorous behavioural responses occurring during a conscious phase. The relative importance of respiratory discomfort versus the potential to induce significant distress due to convulsive wing flapping and associated trauma is a matter for debate. We argue that respiratory discomfort is unpleasant but may be preferable to the risk of vigorous wing flapping and associated injury while conscious in poultry during CAS.</jats:p

An evolutionarily ancient mechanism for regulation of hemoglobin expression in vertebrate red cells

The oxygen transport function of hemoglobin (HB) is thought to have arisen ∼500 million years ago, roughly coinciding with the divergence between jawless (Agnatha) and jawed (Gnathostomata) vertebrates. Intriguingly, extant HBs of jawless and jawed vertebrates were shown to have evolved twice, and independently, from different ancestral globin proteins. This raises the question of whether erythroid-specific expression of HB also evolved twice independently. In all jawed vertebrates studied to date, one of the HB gene clusters is linked to the widely expressed NPRL3 gene. Here we show that the nprl3-linked hb locus of a jawless vertebrate, the river lamprey (Lampetra fluviatilis), shares a range of structural and functional properties with the equivalent jawed vertebrate HB locus. Functional analysis demonstrates that an erythroid-specific enhancer is located in intron 7 of lamprey nprl3, which corresponds to the NPRL3 intron 7 MCS-R1 enhancer of jawed vertebrates. Collectively, our findings signify the presence of an nprl3-linked multiglobin gene locus, which contains a remote enhancer that drives globin expression in erythroid cells, before the divergence of jawless and jawed vertebrates. Different globin genes from this ancestral cluster evolved in the current NPRL3-linked HB genes in jawless and jawed vertebrates. This provides an explanation of the enigma of how, in different species, globin genes linked to the same adjacent gene could undergo convergent evolution

SMARCAD1-mediated active replication fork stability maintains genome integrity.

The stalled fork protection pathway mediated by breast cancer 1/2 (BRCA1/2) proteins is critical for replication fork stability. However, it is unclear whether additional mechanisms are required to maintain replication fork stability. We describe a hitherto unknown mechanism, by which the SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily-A containing DEAD/H box-1 (SMARCAD1) stabilizes active replication forks, that is essential to maintaining resistance towards replication poisons. We find that SMARCAD1 prevents accumulation of 53BP1-associated nucleosomes to preclude toxic enrichment of 53BP1 at the forks. In the absence of SMARCAD1, 53BP1 mediates untimely dissociation of PCNA via the PCNA-unloader ATAD5, causing frequent fork stalling, inefficient fork restart, and accumulation of single-stranded DNA. Although loss of 53BP1 in SMARCAD1 mutants rescues these defects and restores genome stability, this rescued stabilization also requires BRCA1-mediated fork protection. Notably, fork protection-challenged BRCA1-deficient naïve- or chemoresistant tumors require SMARCAD1-mediated active fork stabilization to maintain unperturbed fork progression and cellular proliferation