Cathepsin D non-proteolytically induces proliferation and migration in human omental microvascular endothelial cells via activation of the ERK1/2 and PI3K/AKT pathways

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this recordEpithelial ovarian cancer (EOC) frequently metastasises to the omentum, a process that requires pro-angiogenic activation of human omental microvascular endothelial cells (HOMECs) by tumour-secreted factors. We have previously shown that ovarian cancer cells secrete a range of factors that induce pro-angiogenic responses e.g. migration, in HOMECs including the lysosomal protease cathepsin D (CathD). However, the cellular mechanism by which CathD induces these cellular responses is not understood. The aim of this study was to further examine the pro-angiogenic effects of CathD in HOMECs i.e. proliferation and migration, to investigate whether these effects are dependent on CathD catalytic activity and to delineate the intracellular signalling kinases activated by CathD. We report, for the first time, that CathD significantly increases HOMEC proliferation and migration via a non-proteolytic mechanism resulting in activation of ERK1/2 and AKT. These data suggest that EOC cancer secreted CathD acts as an extracellular ligand and may play an important pro-angiogenic, and thus pro-metastatic, role by activating the omental microvasculature during EOC metastasis to the omentum.FORCE Cancer Charit

High Expression of Galectin-1, VEGF and Increased Microvessel Density are Associated with MELF Pattern in Stage I-III Endometrioid Endometrial Adenocarcinoma

This is the final version. Available on open access via the DOI in this recordBackground: In this study, we investigate the expression of markers of angiogenesis and microvessel density (MVD) in cases of microcystic, elongated and fragmented (MELF) pattern, with its prognostic role in the survival of endometrioid endometrial cancer (EA) patients. Materials and Methods: In this study, 100 cases of EA, 49 cases with MELF pattern and 51 without, were immunohistochemically stained for galectin-1, vascular endothelial growth factor (VEGF), and MVD. Morphometry and statistical (univariate and multivariate) analyses were performed to assess overall survival (OS) and disease-free survival. Results: The expression of VEGF (p<.001) and galectin-1 (p<.001), as well asMVD area (p=.0003) and number of vessels/mm2 (p=.0043), were significantly higher in the +MELF pattern group compared to the -MELF group. A low negative correlation between MELF-pattern and the number of days of survival (p<.001, r=-0.47) was also found. A low positive correlation of MELF-pattern with galectin-1 expression (p<.001, r=0.39), area of vessels/mm2 (p<.001, r=0.36), outcome of EA (p<.001, r = 0.42) and VEGF expression (p<.001, r=0.39) suggests potential pathological relevance of these factors in the prognosis of EA. A univariate survival analysis indicated a role for all parameters of survival. Multivariate Cox proportional hazard regression analysis revealed that only area of vessels/mm2 (hazard ratio [HR], 1.018; 95% confidence interval [CI], 1.002 to 1.033), galectin-1 (HR, 1.049; 95% CI, 1.025 to 1.074) and VEGF (HR, 1.049; 95% CI, 1.022 to 1.077) play key roles in OS. Conclusion: This study reports an increase in MVD, VEGF and galectin-1 expression in EA with MELF pattern and suggests that MELF pattern, along with the angiogenic profile, may be a prognostic factor in EA.FORCE Cancer CharityGomel State Medical Universit

Microstructural Characterisation of Resistance Artery Remodelling in Diabetes Mellitus

This is the final version. Available on open access from Karger Publishers via the DOI in this recordIntroduction: Microvascular remodelling is a symptom of cardiovascular disease. Despite the mechanical environment being recognised as a major contributor to the remodelling process, it is currently only understood in a rudimentary way. Objective: Amorphological and mechanicalevaluation of the resistance vasculature in health and diabetes mellitus.Methods: The cells and extracellular matrix of human subcutaneous resistance arteriesfrom abdominal fat biopsieswere imagedusing two-photon fluorescence and second harmonic generationat varying transmural pressure.The results informed a two-layer mechanical model.Results: Diabetic resistance arteries reducedin wall area as pressure was increased. This was attributed to the presence of thick, straight collagen fibre bundles that bracedthe outer wall.The abnormal mechanical environment caused theinternal elastic lamina and endothelial and vascular smooth muscle cellarrangementsto twist. Conclusions: Our resultssuggest diabetic microvascular remodelling is likely to be stress-driven, comprisingat least two stages: 1. Laying down of adventitial bracing fibres that limit outward distension, and 2. Deposition of additional collagen in the media, likely due to the significantly altered mechanical environment. This work represents a step towards elucidating the local stress environment of cells, which iscrucial to build accurate models of mechanotransduction in disease.British Heart FoundationMedical Research Council (MRC)National Institute for Health Research (NIHR

Hydrogen Sulfide Is a Novel Protector of the Retinal Glycocalyx and Endothelial Permeability Barrier

This is the final version. Available on open access from Frontiers Media via the DOI in this recordData Availability Statement: The original contributions presented in the study are included in the article/supplementary material. The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Significantly reduced levels of the anti-inflammatory gaseous transmitter hydrogen sulfide (H2S) are observed in diabetic patients and correlate with microvascular dysfunction. H2S may protect the microvasculature by preventing loss of the endothelial glycocalyx. We tested the hypothesis that H2S could prevent or treat retinal microvascular endothelial dysfunction in diabetes. Bovine retinal endothelial cells (BRECs) were exposed to normal (NG, 5.5 mmol/L) or high glucose (HG, 25 mmol/L) ± the slow-release H2S donor NaGYY4137 in vitro. Glycocalyx coverage (stained with WGA-FITC) and calcein-labeled monocyte adherence were measured. In vivo, fundus fluorescein angiography (FFA) was performed in normal and streptozotocin-induced (STZ) diabetic rats. Animals received intraocular injection of NaGYY4137 (1 μM) or the mitochondrial-targeted H2S donor AP39 (100 nM) simultaneously with STZ (prevention) or on day 6 after STZ (treatment), and the ratio of interstitial to vascular fluorescence was used to estimate apparent permeability. NaGYY4137 prevented HG-induced loss of BREC glycocalyx, increased monocyte binding to BRECs (p ≤ 0.001), and increased overall glycocalyx coverage (p ≤ 0.001). In rats, the STZ-induced increase in apparent retinal vascular permeability (p ≤ 0.01) was significantly prevented by pre-treatment with NaGYY4137 and AP39 (p < 0.05) and stabilized by their post-STZ administration. NaGYY4137 also reduced the number of acellular capillaries (collagen IV + /IB4-) in the diabetic retina in both groups (p ≤ 0.05). We conclude that NaGYY4137 and AP39 protected the retinal glycocalyx and endothelial permeability barrier from diabetes-associated loss of integrity and reduced the progression of diabetic retinopathy (DR). Hydrogen sulfide donors that target the glycocalyx may therefore be a therapeutic candidate for DR.Medical Research Council (MRC)British Heart FoundationRoyal SocietyBrian Ridge ScholarshipNational Eye Research CentreMasonic Charitable Foundatio

Listening geographies: Landscape, affect and geotechnologies

This paper argues for expanded listening in geography. Expanded listening addresses how bodies of all kinds, human and more-than-human, respond to sound. We show how listening can contribute to research on a wide range of topics, beyond enquiry where sound itself is the primary substantive interest. This is demonstrated through close discussion of what an amplified sonic sensibility can bring to three areas of contemporary geographical interest: geographies of landscape, of affect, and of geotechnologies

RADIUS Authentication in wireless lab environment

The aim of the thesis was to learn and research the development of the Wi-Fi access control methods. This topic will be covered in a chronological order starting from old open-access networks ending to modern methods used today. The practical aim of the study is to implement network access control with an external RADIUS server and to create a RADIUS lab for students. This method has two main advantages. The use of the external server provides a possibility to use one database for many devices like access points and routers. Resources used for this thesis are MB316 classroom access points, hubs, computers and virtual machines of Mikkeli University of Applied Sciences. In the beginning of the study, the theoretical foundation for the study is created with a literature review on authentication, authorization and accounting in Wireless Local Area Networks (WLANs). In the practical part of the study, I implement three different forms of wireless centralized authentication. These forms were autonomous access point RADIUS, Medium business RADIUS setup in Windows server environment and Enterprise RADIUS setup in Windows server environment The findings of the study suggest that autonomous access point solution is no longer a valid method even in small environments because it is not supported in Windows 10 operating system. The server-based solutions either implemented with one or multiple servers are the best for centralized user au-thentication management and access control

Influence of luminescent graphene quantum dots on trypsin activity

Tanveer A Tabish,1,2 Md Zahidul I Pranjol,2 Ilayda Karadag,1 David W Horsell,1 Jacqueline L Whatmore,2 Shaowei Zhang1 1College of Engineering, Mathematics and Physical Sciences, University of Exeter, Exeter, UK; 2Institute of Biomedical and Clinical Science, University of Exeter Medical School, Exeter, UK Background: Protein&ndash;graphene interactions have the potential to play a pivotal role in the future directions of nanomedicine. These interactions lead to diverse processes such as generation of protein coronas, nano&ndash;bio interfaces, particle wrapping, and biocatalytic processes that could determine the ultimate fate of graphene nanocomposites in biologic systems. However, such interactions and their effects on the bioavailability of graphene have not yet been widely appreciated, despite the fact that this is the primary surface in contact with cells.Methods: This paper reports on the integrative physiochemical interaction between trypsin and graphene quantum dots (GQDs) to determine their potential biologic identity in enzyme engineering. This interaction was measured by a wide range of analytical methods.Results: Definitive binding and modulation of trypsin&ndash;GQDs was demonstrated for the first time by use of vibrational spectroscopy and wetting transparency, which revealed that trypsin was absorbed on GQDs&rsquo; surface through its cationic and hydrophilic residues. Our findings suggested that trypsin&rsquo;s active sites were stabilized and protected by the GQDs, which were likely to be responsible for the high bioavailability of GQDs in enzymes.Conclusion: Our work demonstrates the efficacy of GQDs as an enzyme modulator with high specificity, and their great application potential in enzyme engineering as well as enzyme-based therapies. Keywords: graphene, enzyme, luminescence, bioavailability, surface energy&nbsp

Az ASS Berendezési Rendszerek Ipari Bt. vagyoni, pénzügyi és jövedelemi helyzetének elemzése

This is the final version. Available from Dove Medical Press via the DOI in this record.Background: Protein–graphene interactions have the potential to play a pivotal role in the future directions of nanomedicine. These interactions lead to diverse processes such as genera-tion of protein coronas, nano–bio interfaces, particle wrapping, and biocatalytic processes that could determine the ultimate fate of graphene nanocomposites in biologic systems. However, such interactions and their effects on the bioavailability of graphene have not yet been widely appreciated, despite the fact that this is the primary surface in contact with cells. Methods: This paper reports on the integrative physiochemical interaction between trypsin and graphene quantum dots (GQDs) to determine their potential biologic identity in enzyme engineering. This interaction was measured by a wide range of analytical methods. Results: Definitive binding and modulation of trypsin–GQDs was demonstrated for the first time by use of vibrational spectroscopy and wetting transparency, which revealed that trypsin was absorbed on GQDs’ surface through its cationic and hydrophilic residues. Our findings suggested that trypsin’s active sites were stabilized and protected by the GQDs, which were likely to be responsible for the high bioavailability of GQDs in enzymes. Conclusion: Our work demonstrates the efficacy of GQDs as an enzyme modulator with high specificity, and their great application potential in enzyme engineering as well as enzyme-based therapies.EPSRCFORCE Cancer Charit

In vitrotoxic effects of reduced graphene oxide nanosheets on lung cancer cells

journal_title: Nanotechnology article_type: paper article_title: toxic effects of reduced graphene oxide nanosheets on lung cancer cells copyright_information: © 2017 IOP Publishing Ltd date_received: 2017-08-30 date_accepted: 2017-10-24 date_epub: 2017-11-2