Roles and regulation of membrane-associated serine proteases

Pericellular proteolytic activity affects many aspects of cellular behaviour, via mechanisms involving processing of the extracellular matrix, growth factors and receptors. The serine proteases have exquisitely sensitive regulatory mechanisms in this setting, involving both receptor-bound and transmembrane proteases. Receptor-bound proteases are exemplified by the uPA (urokinase plasminogen activator)/uPAR (uPAR receptor) plasminogen activation system. The mechanisms initiating the activity of this proteolytic system on the cell surface, a critical regulatory point, are poorly understood. We have found that the expression of the TTSP (type II transmembrane serine protease) matriptase is highly regulated in leucocytes, and correlates with the presence of active uPA on their surface. Using siRNA (small interfering RNA), we have demonstrated that matriptase specifically activates uPAR-associated pro-uPA. The uPA/uPAR system has been implicated in the activation of the plasminogen-related growth factor HGF (hepatocyte growth factor). However, we find no evidence for this, but instead that HGF can be activated by both matriptase and the related TTSP hepsin in purified systems. Hepsin is of particular interest, as the proteolytic cleavage sequence of HGF is an ‘ideal substrate’ for hepsin and membrane-associated hepsin activates HGF with high efficiency. Both of these TTSPs can be activated autocatalytically at the cell surface, an unusual mechanism among the serine proteases. Therefore these TTSPs have the capacity to be true upstream initiators of proteolytic activity with subsequent downstream effects on cell behaviour

IGF-1 receptor activity in the Golgi of migratory cancer cells depends on adhesion-dependent phosphorylation of Tyr1250 and Tyr1251

Although insulin-like growth factor 1 (IGF-1) signaling promotes tumor growth and cancer progression, therapies that target the IGF-1 receptor (IGF-1R) have shown poor clinical efficacy. To address IGF-1R activity in cancer cells and how it differs from that of the closely related insulin receptor (IR), we focused on two tyrosines in the IGF-1R C-terminal tail that are not present in the IR and are essential for IGF-1–mediated cancer cell survival, migration, and tumorigenic growth. We found that Tyr1250 and Tyr1251 (Tyr1250/1251) were autophosphorylated in a cell adhesion–dependent manner. To investigate the consequences of this phosphorylation, we generated phosphomimetic Y1250E/Y1251E (EE) and nonphosphorylatable Y1250F/Y1251F (FF) mutant forms of IGF-1R. Although fully competent in kinase activity and signaling, the EE mutant was more rapidly internalized and degraded than either the wild-type or FF receptor. IGF-1 promoted the accumulation of wild-type and EE IGF-1R within the Golgi apparatus, whereas the FF mutant remained at the plasma membrane. Golgi-associated IGF-1R signaling was a feature of migratory cancer cells, and Golgi disruption impaired IGF-1–induced signaling and cell migration. Upon the formation of new cell adhesions, IGF-1R transiently relocalized to the plasma membrane from the Golgi. Thus, phosphorylation at Tyr1250/1251 promoted IGF-1R translocation to and signaling from the Golgi to support an aggressive cancer phenotype. This process distinguishes IGF-1R from IR signaling and could contribute to the poor clinical efficacy of antibodies that target IGF-1R on the cell surface

Checklists and illustrative financial statements for life insurance companies : a financial accounting and reporting practice aid, April 1992 edition

https://egrove.olemiss.edu/aicpa_indev/1878/thumbnail.jp

Ozone profile retrievals from the ESA GOME instrument

The potential of the ESA Global Ozone Monitoring Experiment (GOME) to produce ozone profile information has been examined by carrying out two sample retrievals using simulated GOME data. The first retrieval examines the potential of the GOME instrument to produce stratospheric ozone profiles using the traditional back-scatter ultraviolet technique, while the second examines the possibility of obtaining tropospheric profile information, and improving the quality of the stratospheric profile retrievals, by exploiting the temperature dependence of the ozone Huggins bands

Co-research with older people:perspectives on impact

Although research in partnership with older people has been slower to develop than that with some other service user ‘groups’, there are a growing number of health and social care studies that have involved older people as co-researchers. We build on this existing evidence by addressing two key areas. First, despite the growth of participatory research with older people generally, some groups tend to be excluded. We focus particularly on a project in which older people with dementia and older people from a black and minority ethnic community were involved as co-researchers. They worked with academic researchers in all stages of the research process, exploring other older people’s experiences of transitions between care services. Second, recent literature suggests a lack of critical evaluation of involvement, arguing that researchers tend to emphasise the positives on the basis of retrospective narrative accounts of the process, rather than critically appraising the impact of involvement. This article offers a critical account of the impact of a participatory approach at different stages of a research project, evaluating this from the perspective of different stakeholders. In so doing, we engage with incisive critiques which claim that, far from empowering service users, much service user involvement activity contributes to their oppression. We conclude that participatory research with marginalised older people has the potential to achieve meaningful change at both individual and social levels. However, in view of its dangers and limitations, we argue the need for the impact of participatory research to be carefully evaluated from the perspectives of all parties in the process. </jats:p

Early Contrast Enhancement: a novel Magnetic Resonance Imaging biomarker of pleural malignancy

Introduction: Pleural Malignancy (PM) is often occult on subjective radiological assessment. We sought to define a novel, semi-objective Magnetic Resonance Imaging (MRI) biomarker of PM, targeted to increased tumour microvessel density (MVD) and applicable to minimal pleural thickening. Materials and methods: 60 consecutive patients with suspected PM underwent contrast-enhanced 3-T MRI then pleural biopsy. In 58/60, parietal pleura signal intensity (SI) was measured in multiple regions of interest (ROI) at multiple time-points, generating ROI SI/time curves and Mean SI gradient (MSIG: SI increment/time). The diagnostic performance of Early Contrast Enhancement (ECE; which was defined as a SI peak in at least one ROI at or before 4.5 min) was compared with subjective MRI and Computed Tomography (CT) morphology results. MSIG was correlated against tumour MVD (based on Factor VIII immunostain) in 31 patients with Mesothelioma. Results: 71% (41/58) patients had PM. Pleural thickening was &#60;10 mm in 49/58 (84%). ECE sensitivity was 83% (95% CI 61–94%), specificity 83% (95% CI 68–91%), positive predictive value 68% (95% CI 47–84%), negative predictive value 92% (78–97%). ECE performance was similar or superior to subjective CT and MRI. MSIG correlated with MVD (r = 0.4258, p = .02). Discussion: ECE is a semi-objective, perfusion-based biomarker of PM, measurable in minimal pleural thickening. Further studies are warranted

Multiple mechanisms for enhanced plasmodesmata density in disparate subtypes of C4 grasses

Proliferation of plasmodesmata (PD) connections between bundle sheath (BS) and mesophyll (M) cells has been proposed as a key step in the evolution of two-cell C4 photosynthesis; However, a lack of quantitative data has hampered further exploration and validation of this hypothesis. In this study, we quantified leaf anatomical traits associated with metabolite transport in 18 species of BEP and PACMAD grasses encompassing four origins of C4 photosynthesis and all three C4 subtypes (NADP-ME, NAD-ME, and PCK). We demonstrate that C4 leaves have greater PD density between M and BS cells than C3 leaves. We show that this greater PD density is achieved by increasing either the pit field (cluster of PD) area or the number of PD per pit field area. NAD-ME species had greater pit field area per M–BS interface than NADP-ME or PCK species. In contrast, NADP-ME and PCK species had lower pit field area with increased number of PD per pit field area than NAD-ME species. Overall, PD density per M–BS cell interface was greatest in NAD-ME species while PD density in PCK species exhibited the largest variability. Finally, the only other anatomical characteristic that clearly distinguished C4 from C3 species was their greater Sb value, the BS surface area to subtending leaf area ratio. In contrast, BS cell volume was comparable between the C3 and C4 grass species examined.FRD is supported by scholarship awards from the Lee Foundation (IRRI) and the Australian Research Council Centre of Excellence for Translational Photosynthesis (CE140100015). SK is a Royal Society University Research Fellow. Work in SK’s lab is supported by the European Union’s Horizon 2020 research and innovation programme under grant agreement number 637765

Micro-enterprises: small enough to care?

This report presents findings of an evaluation of micro-enterprises in social care in England, which ran from 2013 to 2015. Organisations are here classed as micro if they employ five or fewer full-time equivalent staff. The aim of the project was to test the extent to which micro-enterprises deliver services that are personalised, valued, innovative and cost-effective, and how they compare with small, medium and large providers. Working in three parts of the country, researchers compared 27 organisations providing care and support, of which 17 were microenterprises, 2 were small, 4 were medium and 4 were large. The project team interviewed and surveyed 143 people (staff, older people, people with disabilities and carers) who received support from the 27 providers. The findings presented are relevant to people who use services and their families; social care commissioners; regulators and policy makers at a local and national level; people who provide care services; and social entrepreneurs who are considering setting up micro forms of support. The research was based at the University of Birmingham. It was funded by the Economic and Social Research Council (ESRC), as part of a project entitled Does Smaller mean Better? Evaluating Micro-enterprises in Adult Social Care (ESRC Standard Grant ES/ K002317/1)