Inhibition of thiol isomerase activity diminishes endothelial activation of plasminogen, but not of protein C

Highlights •A range of thiol isomerase enzymes were expressed by HMEC-1 endothelial cells. •Inhibition of thiol isomerases reduced plasminogen activation on HMEC-1 cells. •Exogenous bovine protein disulphide isomerase increased plasminogen activation in HMEC-1 cells. •Inhibition of thiol isomerases also reduced fibrin clot lysis, but not via direct effects on tPA. •HMEC-1 mediated protein C activation was unaffected by thiol isomerase inhibition. Abstract Introduction Cell surface thiol isomerase enzymes, principally protein disulphide isomerase (PDI), have emerged as important regulators of platelet function and tissue factor activation via their action on allosteric disulphide bonds. Allosteric disulphides are present in other haemostasis-related proteins, and we have therefore investigated whether thiol isomerase inhibition has any influence on two endothelial activities relevant to haemostatic regulation, namely activation of protein C and activation of plasminogen, with subsequent fibrinolysis. Materials and Methods The study was performed using the human microvascular endothelial cell line HMEC-1. Thiol isomerase gene expression was measured by RT-PCR and activation of protein C and plasminogen by cell-based assays using chromogenic substrates S2366 and S2251, respectively. Cell mediated fibrinolysis was measured by monitoring absorbance at 405 nm following fibrin clot formation on the surface of HMEC-1 monolayers. Results and Conclusions A variety of thiol isomerase enzymes, including PDI, were expressed by HMEC-1 cells and thiol reductase activity detectable on the cell surface was inhibited by both RL90 anti-PDI antibody and by the PDI inhibitor quercetin-3-rutinoside (rutin). In cell-based assays, activation of plasminogen, but not of protein C, was inhibited by RL90 antibody and, to a lesser extent, by rutin. Fibrin clot lysis occurring on a HMEC-1 monolayer was also significantly slowed by RL90 antibody and by rutin, but RL90-mediated inhibition was abolished in the presence of exogenous tissue plasminogen activator (tPA). We conclude that thiol isomerases, including PDI, are involved in fibrinolytic regulation at the endothelial surface, although not via a direct action on tPA. These findings broaden understanding of haemostatic regulation by PDI, and may aid in development of novel anti-thrombotic therapeutic strategies targeted via the fibrinolysis system

Security and privacy challenges in smart cities

© 2018 Elsevier Ltd The construction of smart cities will bring about a higher quality of life to the masses through digital interconnectivity, leading to increased efficiency and accessibility in cities. Smart cities must ensure individual privacy and security in order to ensure that its citizens will participate. If citizens are reluctant to participate, the core advantages of a smart city will dissolve. This article will identify and offer possible solutions to five smart city challenges, in hopes of anticipating destabilizing and costly disruptions. The challenges include privacy preservation with high dimensional data, securing a network with a large attack surface, establishing trustworthy data sharing practices, properly utilizing artificial intelligence, and mitigating failures cascading through the smart network. Finally, further research directions are provided to encourage further exploration of smart city challenges before their construction

The structure and dynamics of young star clusters: King 16, NGC 1931, NGC 637 and NGC 189

In this paper, using 2MASS photometry, we study the structural and dynamical properties of four young star clusters viz. King 16, NGC 1931, NGC 637 and NGC 189. For the clusters King 16, NGC 1931, NGC 637 and NGC 189, we obtain the limiting radii of 7', 12', 6' and 5' which correspond to linear radii of 3.6 pc, 8.85 pc, 3.96 pc and 2.8 pc respectively. The reddening values $E(B-V)$ obtained for the clusters are 0.85, 0.65--0.85, 0.6 and 0.53 and their true distances are 1786 pc, 3062 pc, 2270 pc and 912 pc respectively. Ages of the clusters are 6 Myr, 4 Myr, 4 Myr and 10 Myr respectively. We compare their structures, luminosity functions and mass functions ($\phi(M) = dN/dM \propto M^{-(1+\chi)}$) to the parameter $\tau = t_{age}/t_{relax}$ to study the star formation process and the dynamical evolution of these clusters. We find that, for our sample, mass seggregation is observed in clusters or their cores only when the ages of the clusters are comparable to their relaxation times ($\tau \geq 1$). These results suggest mass seggregation due to dynamical effects. The values of $\chi$, which characterise the overall mass functions for the clusters are 0.96 $\pm$ 0.11, 1.16 $\pm$ 0.18, 0.55 $\pm$ 0.14 and 0.66 $\pm$ 0.31 respectively. The change in $\chi$ as a function of radius is a good indicator of the dynamical state of clusters.Comment: Accepted for publication in Astrophysics & Space Scienc

Critical current density and flux pinning in La2-xPrxCa2xBa2Cu4+2xOz (x = 0.1 - 0.5) superconductors

Polycrystalline La2-xPrxCa2xBa2Cu4+2xOz (LPCaBCO) compounds with x = 0.1 - 0.5 were synthesized by solid-state reaction method and studied by room temperature X-ray diffraction, dc resistivity, dc magnetization and iodometry. The superconducting transition temperatures in these tetragonal triple perovskite compounds increases from 32 to 62 K (Tconset values) with increasing dopant concentration. The mixing of rare earth La3+ and Pr3+/4+ ions at rare earth site (La3+) along with substitution of divalent Ca2+ results in the shrinkage of unit cell volume. The contraction of unit cell volume due to larger ion being substituted by smaller ions, gives rise to creation of pinning centers in the unit cell leading to increase in critical current density and flux pinningComment: To be published in Solid State Communications (2004

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Prelamin A mediates myocardial inflammation in dilated and HIV-associated cardiomyopathies

Cardiomyopathies are complex heart muscle diseases that can be inherited or acquired. Dilated cardiomyopathy can result from mutations in LMNA, encoding the nuclear intermediate filament proteins lamin A/C. Some LMNA mutations lead to accumulation of the lamin A precursor, prelamin A, which is disease causing in a number of tissues, yet its impact upon the heart is unknown. Here, we discovered myocardial prelamin A accumulation occurred in a case of dilated cardiomyopathy, and we show that a potentially novel mouse model of cardiac-specific prelamin A accumulation exhibited a phenotype consistent with inflammatory cardiomyopathy, which we observed to be similar to HIV-associated cardiomyopathy, an acquired disease state. Numerous HIV protease therapies are known to inhibit ZMPSTE24, the enzyme responsible for prelamin A processing, and we confirmed that accumulation of prelamin A occurred in HIV+ patient cardiac biopsies. These findings (a) confirm a unifying pathological role for prelamin A common to genetic and acquired cardiomyopathies; (b) have implications for the management of HIV patients with cardiac disease, suggesting protease inhibitors should be replaced with alternative therapies (i.e., nonnucleoside reverse transcriptase inhibitors); and (c) suggest that targeting inflammation may be a useful treatment strategy for certain forms of inherited cardiomyopathy

Anesthesia considerations for cataract surgery in patients with schizophrenia: A narrative review

Schizophrenia is ranked among the top 10 global burdens of disease. About 1 of people meet the diagnostic criteria for this disorder over their lifetime. Schizophrenic patients can develop cataract, particularly related to age and medications, requiring surgery and anesthesia. Many concerning factors, including cognitive function, anxiety, behavioral issues, poor cooperation and paroxysmal movements, may lead to general anesthesia as the default method. Antipsychotic agents should be continued during the periop-erative period if possible. Topical/regional anesthesia is suitable in most schizophrenic patients undergoing cataract surgery. It reduces potential drug interactions and many postoperative complications; however, appropriate patient selection is paramount to its success. General anesthesia remains the primary technique for patients who are considered unsuitable for the topical/regional technique. Early involvement of a psychiatrist in the perioperative period, especially for patients requiring general anesthesia, is beneficial but often under-utilized. This narrative review summarizes the anesthetic considerations for cataract surgery in patients with schizophrenia. Keywords. © 2021, Author(s)

Tomato: a crop species amenable to improvement by cellular and molecular methods

Tomato is a crop plant with a relatively small DNA content per haploid genome and a well developed genetics. Plant regeneration from explants and protoplasts is feasable which led to the development of efficient transformation procedures. In view of the current data, the isolation of useful mutants at the cellular level probably will be of limited value in the genetic improvement of tomato. Protoplast fusion may lead to novel combinations of organelle and nuclear DNA (cybrids), whereas this technique also provides a means of introducing genetic information from alien species into tomato. Important developments have come from molecular approaches. Following the construction of an RFLP map, these RFLP markers can be used in tomato to tag quantitative traits bred in from related species. Both RFLP's and transposons are in the process of being used to clone desired genes for which no gene products are known. Cloned genes can be introduced and potentially improve specific properties of tomato especially those controlled by single genes. Recent results suggest that, in principle, phenotypic mutants can be created for cloned and characterized genes and will prove their value in further improving the cultivated tomato.

Interactions between cell surface protein disulphide isomerase and S-nitrosoglutathione during nitric oxide delivery

In this study, we investigated the role of protein disulphide isomerase (PDI) in rapid metabolism of S-nitrosoglutathione (GSNO) and S-nitrosoalbumin (albSNO) and in NO delivery from these compounds into cells. Incubation of GSNO or albSNO (1 μM) with the megakaryocyte cell line MEG-01 resulted in a cell-mediated removal of each compound which was inhibited by blocking cell surface thiols with 5,5′-dithiobis 2-nitrobenzoic acid (DTNB) (100 μM) or inhibiting PDI with bacitracin (5 mM). GSNO, but not albSNO, rapidly inhibited platelet aggregation and stimulated cyclic GMP (cGMP) accumulation (used as a measure of intracellular NO entry). cGMP accumulation in response to GSNO (1 μM) was inhibited by MEG-01 treatment with bacitracin or DTNB, suggesting a role for PDI and surface thiols in NO delivery. PDI activity was present in MEG-01 conditioned medium, and was inhibited by high concentrations of GSNO (500 μM). A number of cell surface thiol-containing proteins were labelled using the impermeable thiol specific probe 3-(N-maleimido-propionyl) biocytin (MPB). Pretreatment of cells with GSNO resulted in a loss of thiol reactivity on some but not all proteins, suggesting selective cell surface thiol modification. Immunoprecipitation experiments showed that GSNO caused a concentration-dependent loss of thiol reactivity of PDI. Our data indicate that PDI is involved in both rapid metabolism of GSNO and intracellular NO delivery and that during this process PDI is itself altered by thiol modification. In contrast, the relevance of PDI-mediated albSNO metabolism to NO signalling is uncertain