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Atherosclerosis is commonly known as an inflammatory disease that is characterized by lipid deposition in the arterial wall, causing gradual restriction or complete blockade of blood flow, which can cause complications such as myocardial infarction, stroke, or peripheral artery disease. Several factors contribute to initiation and progression of atherosclerotic plaque formation. The role of macrophages and leukocytes in atherosclerosis has been well explored. Here, we provide an overview of what has been reported on the role and impact of the arterial cells on plaque formation, and vice versa. The atherogenic environment can trigger transformation and dedifferentiation of the endothelial cells (ECs), smooth muscle cells, and fibroblasts (FBs) whereby they can either directly contribute to plaque formation or influence its composition. Recent studies have demonstrated the plasticity in the identity of the arterial cells, the formation of intermediate cell types that share the characteristics of multiple cell types, and have revealed novel roles and functions for these cells in atherosclerosis. The potential for all vascular cells to cross-transdifferentiate, and detection of cells with mosaic characteristics in the atherosclerotic plaques reveal that the plaque environment is a complex and dynamic environment that could regulate the disease progression independent from the circulating lipid levels. We will also provide an overview on the interplay between sex and atherosclerosis, which has remained an underexplored area. [ABSTRACT FROM AUTHOR]

Sampling of environmental DNA (eDNA) and high‐throughput sequencing is emerging as an effective biodiversity assessment method in aquatic systems and may have utility for biodiversity inventory in terrestrial environments. Rock pools (gnammas) on granite inselbergs support a rich community of aquatic organisms and are culturally important to Indigenous peoples worldwide. However, the application of eDNA to survey rock pool biodiversity are undocumented. In a collaborative study with traditional owners, we explore the application of eDNA metabarcoding and traditional sampling methods to document and compare species richness and composition of eukaryotes from 15 rock pools (pits and pans) on granite inselbergs in southeastern Australia. We detected 116 taxonomic units, 81 assignments from eDNA sequencing and 35 species (23 rotifers and 12 microcrustaceans) using microscopic analysis. eDNA detected a broad range of taxa not previously documented from rock pools in Australia, although significantly more zooplankton (rotifers and microcrustaceans) were detected under a microscope, including several rare species and two undescribed species of rotifer. Brachionus calyciflorus and B. angularis were the only rotifer species assigned to species level through eDNA sequencing and were detected using both methods. We found no significant difference in mean species richness between rock pool types; however, species composition differed significantly between pits and pans. This study highlights the value of using eDNA to document biodiversity of ephemeral aquatic habitats in terrestrial ecosystems but reveals the general lack of reference sequence data for microorganisms, underscoring the value of using traditional sampling and taxonomic assignment methods. [ABSTRACT FROM AUTHOR]

Sustained, indolent immune injury of the vasculature of a heart transplant limits long-term graft and recipient survival. This injury is mitigated by a poorly characterized, maladaptive repair response. Vascular endothelial cells respond to proangiogenic cues in the embryo by differentiation to specialized phenotypes, associated with expression of apelin. In the adult, the role of developmental proangiogenic cues in repair of the established vasculature is largely unknown. We found that human and minor histocompatibility-mismatched donor mouse heart allografts with alloimmune-mediated vasculopathy upregulated expression of apelin in arteries and myocardial microvessels. In vivo, loss of donor heart expression of apelin facilitated graft immune cell infiltration, blunted vascular repair, and worsened occlusive vasculopathy in mice. In vitro, an apelin receptor agonist analog elicited endothelial nitric oxide synthase activation to promote endothelial monolayer wound repair and reduce immune cell adhesion. Thus, apelin acted as an autocrine growth cue to sustain vascular repair and mitigate the effects of immune injury. Treatment with an apelin receptor agonist after vasculopathy was established markedly reduced progression of arterial occlusion in mice. Together, these initial data identify proangiogenic apelin as a key mediator of coronary vascular repair and a pharmacotherapeutic target for immune-mediated injury of the coronary vasculature., Introduction Heart transplantation is the most effective treatment of end-stage heart failure to prolong life. Modern immune-suppression regimens blunt alloreactive immune responses against the transplant, but are nevertheless associated with [...]

FYVE domains are highly conserved protein modules that typically bind phosphatidylinositol 3-phosphate (PI3P) on the surface of early endosomes. Along with pleckstrin homology (PH) and phox homology (PX) domains, FYVE domains are the principal readers of the phosphoinositide (PI) code that mediate specific recognition of eukaryotic organelles. Of all the human FYVE domain containing proteins, those within the faciogenital dysplasia (Fgd) subfamily are particularly divergent and couple with GTPases to exert unique cellular functions. The subcellular distributions and functions of these evolutionarily conserved signal transducers, which also include Dbl homology (DH) and two PH domains, are discussed here to better understand the biological range of processes that such multidomain proteins engage in. Determinants of their various functions include specific multidomain architectures, posttranslational modifications including PIP stops that have been discovered in sorting nexins, PI recognition motifs, and phospholipid-binding surfaces as defined by the Membrane Optimal Docking Area (MODA) program. How these orchestrate Fgd function remains unclear but has implications for developmental diseases including Aarskog-Scott syndrome, which is also known as faciogenital dysplasia, and forms of cancer that are associated with mutations and amplifications of Fgd genes. Key words: Cdc42, Dbl, FYVE, Fgd, GEF, GTPase, PH, phosphoinositide, pleckstrin, cancer, faciogenital dysplasia, Aarskog-Scott syndrome, lipid signaling, membrane trafficking, MODA, PIP, Rho. Les domaines FYVE consistent en modules proteiques hautement conserves qui lient typiquement le phosphatidylinositol 3-phosphate (PI3P) a la surface des endosomes precoces. Avec les domaines homologues de la pleckstrine (PH) et de phox (PX), les domaines FYVE sont les principaux lecteurs du code des phosphoinositides (PI) qui medient la reconnaissance specifique des organites eucaryotes. De toutes les proteines humaines a domaine FYVE, celles qui appartiennent a la sous-famille Fgd (<< faciogenital dysplasia >>) sont particulierement divergentes et s'associent a des GTPases pour exercer des fonctions cellulaires uniques. Les distributions subcellulaires et les fonctions de ces transducteurs de signaux conserves dans l'evolution, qui comprennent aussi les proteines a domaines d'homologie a Dbl (DH) et a deux domaines PH, sont discutees ici afin de mieux comprendre le spectre biologique des processus dans lesquels de telles proteines a domaines multiples sont engagees. Les determinants de leurs diverses fonctions comprennent des architectures multidomaines specifiques, des modifications post-traductionnelles dont les << PIP stops >> decouverts chez les sorting nexins, des motifs de reconnaissance du PI et des surfaces de liaison des phospholipides tels que definis par le programme << Membrane Optimal Docking Area >> (MODA). L'on ignore encore comment ils orchestrent les fonctions des Fgd, mais ils sont impliques dans des maladies developpementales comme le syndrome Aarskog-Scott, connu aussi sous l'appellation de dysplasie faciogenitale, et dans des cancers associes a des mutations et des amplifications des genes Fgd. [Traduit par la Redaction] Mots-cles : Cdc42, Dbl, FYVE, Fgd, GEF, GTPase, PH, phosphoinositide, pleckstrine, cancer, dysplasie faciogenitale, syndrome Aarskog-Scott, signalisation lipidique, trafic membranaire, MODA, PIP, Rho., Phosphoinositide code Stimulation and differentiation of cells involve dramatic alterations of phosphorylation patterns in proteins, lipids, and nucleotides. Defining how these molecular events converge to change cellular behaviour remains a [...]

Lymphocyte recruitment to sites of inflammation involves a bidirectional series of cues between the endothelial cell (EC) and the leukocyte that culminate in lymphocyte migration into the tissue. Remodeling of the EC F-actin cytoskeleton has been observed after leukocyte adhesion, but the signals to the EC remain poorly defined. We studied the dependence of peripheral blood lymphocyte transendothelial migration (TEM) through an EC monolayer in vitro on EC phosphatidylinositol 3-kinase (PI 3-kinase) activity. Lymphocytes were perfused over cytokine-activated EC using a parallel-plate laminar flow chamber. Inhibition of EC PI 3-kinase activity using LY-294002 or wortmannin decreased lymphocyte TEM (48 [+ or -] 6 or 34 [+ or -] 7%, respectively, vs. control; mean [+ or -] SE; P < 0.05). Similarly, EC knockdown of the p85[alpha] regulatory subunit of PI 3-kinase decreased lymphocyte transmigration. Treatment of EC with jasplakinolide to inhibit EC F-actin remodeling also decreased lymphocyte TEM to 24 [+ or -] 10% vs. control (P < 0.05). EC PI 3-kinase inhibition did not change the strength of lymphocyte adhesion to the EC or formation of the EC 'docking structure' after intercellular adhesion molecule-1 ligation, whereas this was inhibited by jasplakinolide treatment. A similar fraction of lymphocytes migrated on control or LY-294002-treated EC and localized to interendothelial junctions. However, lymphocytes failed to extend processes below the level of vascular endothelial (VE)-cadherin on LY-294002-treated EC. Together these observations indicate that EC PI 3-kinase activity and F-actin remodeling are required during lymphocyte diapedesis and identify a PI 3-kinase-dependent step following initial separation of the VE-cadherin barrier. T lymphocyte; cell motility; adherens junctions; cytoskeleton; F-actin

Analysis of atrial rhythm is important in the treatment and management of patients with atrial fibrillation. Several algorithms exist for extracting the atrial signal from the electrocardiogram (ECG) in atrial fibrillation, but there are few reports on how well these techniques are able to recover the atrial signal. We assessed and compared three algorithms for extracting the atrial signal from the 12-lead ECG. The 12-lead ECGs of 30 patients in atrial fibrillation were analyzed. Atrial activity was extracted by three algorithms, Spatiotemporal QRST cancellation (STC), principal component analysis (PCA), and independent component analysis (ICA). The amplitude and frequency characteristics of the extracted atrial signals were compared between algorithms and against reference data. Mean (standard deviation) amplitude of QRST segments of VI was 0.99 (0.54) mV, compared to 0.18 (0.11) mV (STC), 0.19 (0.13) mV (PCA), and 0.29 (0.22) mV (ICA). Hence, for all algorithms there were significant reductions in the amplitude of the ventricular activity compared with that in VI. Reference atrial signal amplitude in VI was 0.18 (0.11) mV, compared to 0.17 (0.10) mV (STC), 0.12 (0.09) mV (PCA), and 0.18 (0.13) mV (ICA) in the extracted atrial signals. PCA tended to attenuate the atrial signal in these segments. There were no significant differences for any of the algorithms when comparing the amplitude of the reference atrial signal with that of the extracted atrial signals in segments in which ventricular activity had been removed. There were no significant differences between algorithms in the frequency characteristics of the extracted atrial signals. There were discrepancies in amplitude and frequency characteristics of the atrial signal in only a few cases resulting from notable residual ventricular activity for PCA and ICA algorithms. In conclusion, the extracted atrial signals from these algorithms exhibit very similar amplitude and frequency characteristics. Users of these algorithms should be observant of residual ventricular activities which can affect the analysis of the fibrillatory waveform in clinical practice. Index Terms--Atrial fibrillation, atrial signal, comparative study, independent component analysis, principal component analysis, spatiotemporal QRST cancellation.

Split-thickness pig skin was transplanted on severe combined immunodeficient mice so that pig dermal microvessels spontaneously inosculated with mouse microvessels and functioned to perfuse the grafts. Pig endothelial cells in the healed grafts constitutively expressed class I and class II major histocompatibility complex molecules. Major histocompatibility complex molecule expression could be further increased by intradermal injection of pig interferon-[Gamma] (IFN-[Gamma]) but not human IFN-[Gamma] or tumor necrosis factor. Grafts injected with pig IFN-[Gamma] also developed a sparse infiltrate of mouse neutrophils and eosinophils without evidence of injury. Introduction of human peripheral blood mononuclear cells into the animals by intraperitoneal inoculation resulted in sparse perivascular mononuclear cell infiltrates in the grafts confined to the pig dermis. Injection of pig skin grafts on mice that received human peripheral blood mononuclear cells with pig IFN-[Gamma] (but not human IFN-[Gamma] or heat-inactivated pig IFN-[Gamma]) induced human CD[4.sup.+] and CD[8.sup.+] T cells and macrophages to more extensively infiltrate the pig skin grafts and injure pig dermal microvessels. These findings suggest that human T cell-mediated rejection of xenotransplanted pig organs may be prevented if cellular sources of pig interferon (e.g., passenger lymphocytes) are eliminated from the graft.

Rho proteins are signalling molecules that control cellular dynamics, movement and morphological changes. They are activated by Rho guanine-nucleotide exchange factors (Rho GEFs) that transduce upstream signals into Rho-mediated activation of downstream processes. Fgd5 is a Rho GEF involved in angiogenesis and its target Rho protein for this process has been linked to Cdc42 activation. Here, we examined the function of purified Fgd5, specifically, which Rho proteins it activates and pinpoint the structural domains required for enzymatic activity. Using a GEF enzyme assay, we found that purified Fgd5 showed preferential activation of Rac1 and direct binding of Rac1 in pull-down and co-immunoprecipitation assays. Structural comparisons showed that the Fgd5 DH domain is highly similar to the Rac1 GEF, TrioN, supporting a role for Fgd5 as a Rac1 GEF. Compounds that bind to purified Fgd5 DH-PH protein were identified by screening a small molecule library via surface plasmon resonance. The effects of eleven ligands were further examined for their ability to inhibit the Fgd5 GEF enzymatic activity and Rac1 interaction. From these studies, we found that the compound aurintricarboxylic acid, and to a lesser extent mitoxantrone dihydrochloride, inhibited both Fgd5 GEF activation of Rac1 and their interaction. Aurintricarboxylic acid had no effect on the activity or binding of the Rac1 GEF, TrioN, thus demonstrating the feasibility of selectively disrupting Rho GEF activators. Abbreviations: a.a.: amino acid; ATA: aurintricarboxylic acid; DH: Dbl homology; DOCK: dictator of cytokinesis; Fgd: faciogenital dysplasia; GEF: guanine-nucleotide exchange factor; GST: glutathione S-transferase; LOPAC: library of pharmacologically active compounds; PH: pleckstrin homology; PDB: protein data bank; s.e.m.: standard error of the mean; SPR: surface plasmon resonance. [ABSTRACT FROM AUTHOR]