Hepatoma cell density promotes claudin-1 and scavenger receptor BI expression and hepatitis C virus internalization.

Hepatitis C virus (HCV) entry occurs via a pH- and clathrin-dependent endocytic pathway and requires a number of cellular factors, including CD81, the tight-junction proteins claudin 1 (CLDN1) and occludin, and scavenger receptor class B member I (SR-BI). HCV tropism is restricted to the liver, where hepatocytes are tightly packed. Here, we demonstrate that SR-BI and CLDN1 expression is modulated in confluent human hepatoma cells, with both receptors being enriched at cell-cell junctions. Cellular contact increased HCV pseudoparticle (HCVpp) and HCV particle (HCVcc) infection and accelerated the internalization of cell-bound HCVcc, suggesting that the cell contact modulation of receptor levels may facilitate the assembly of receptor complexes required for virus internalization. CLDN1 overexpression in subconfluent cells was unable to recapitulate this effect, whereas increased SR-BI expression enhanced HCVpp entry and HCVcc internalization, demonstrating a rate-limiting role for SR-BI in HCV internalization

Emerging role of LRRK2 in human neural progenitor cell cycle progression, survival and differentiation

Despite a comprehensive mapping of the Parkinson's disease (PD)-related mRNA and protein leucine-rich repeat kinase 2 (LRRK2) in the mammalian brain, its physiological function in healthy individuals remains enigmatic. Based on its structural features and kinase properties, LRRK2 may interact with other proteins involved in signalling pathways. Here, we show a widespread LRRK2 mRNA and/or protein expression in expanded or differentiated human mesencephalic neural progenitor cells (hmNPCs) and in post-mortem substantia nigra PD patients. Using small interfering RNA duplexes targeting LRRK2 in hmNPCs following their differentiation into glia and neurons, we observed a reduced number of dopaminergic neurons due to apoptosis in LRRK2 knockdown samples. LRRK2-deficient hmNPCs exhibited elevated cell cycle- and cell death-related markers. In conclusion, a reduction of LRRK2 expression in hmNPCs severely impaired dopaminergic differentiation and/or survival of dopaminergic neurons most likely via preserving or reactivating the cell cycle

Endothelial-monocyte activating polypeptide II disrupts alveolar epithelial type II to type I cell transdifferentiation

<p>Abstract</p> <p>Background</p> <p>Distal alveolar morphogenesis is marked by differentiation of alveolar type (AT)-II to AT-I cells that give rise to the primary site of gas exchange, the alveolar/vascular interface. Endothelial-Monocyte Activating Polypeptide (EMAP) II, an endogenous protein with anti-angiogenic properties, profoundly disrupts distal lung neovascularization and alveolar formation during lung morphogenesis, and is robustly expressed in the dysplastic alveolar regions of infants with Bronchopulmonary dysplasia. Determination as to whether EMAP II has a direct or indirect affect on ATII→ATI trans-differentiation has not been explored.</p> <p>Method</p> <p>In a controlled nonvascular environment, an <it>in vitro </it>model of ATII→ATI cell trans-differentiation was utilized to demonstrate the contribution that one vascular mediator has on distal epithelial cell differentiation.</p> <p>Results</p> <p>Here, we show that EMAP II significantly blocked ATII→ATI cell transdifferentiation by increasing cellular apoptosis and inhibiting expression of ATI markers. Moreover, EMAP II-treated ATII cells displayed myofibroblast characteristics, including elevated cellular proliferation, increased actin cytoskeleton stress fibers and Rho-GTPase activity, and increased nuclear:cytoplasmic volume. However, EMAP II-treated cells did not express the myofibroblast markers desmin or αSMA.</p> <p>Conclusion</p> <p>Our findings demonstrate that EMAP II interferes with ATII → ATI transdifferentiation resulting in a proliferating non-myofibroblast cell. These data identify the transdifferentiating alveolar cell as a possible target for EMAP II's induction of alveolar dysplasia.</p

Cost evaluation of an innovative outreach clinic across social insurance codes in rural Northern Germany - A routine data analysis

A community outreach clinic was established in 2013 in a rural part of Germany to counsel and better integrate patients with mental health disorders or clients with psychosocial problems into the healthcare and complementary social assistance system. In a pilot study, we aimed to evaluate the costs of the integration assistance system after the outreach clinic was opened, the number of visits and the trend in the costs of the social assistance system of the federal state. Anonymised secondary cost data were used to evaluate the costs associated with the integration assistance receivers before (2010–2012) and after the establishment of the outreach clinic (2013–2015). Total costs were descriptively compared between the intervention group (consultation in the outreach clinic), the non-referral group, and a propensity score-matched control group for the years 2013–2015. To monitor the counselling activity, we used anonymised data on visits to the outreach clinic between 2013 and 2015. Data from 50 clients in the outreach clinic and 678 non-referral clients were analysed. The total costs of the integration assistance for the years 2013–2015 amounted to EUR 21,516 (95% CI 14,513–28,518) and EUR 28,464 (25,789–31,140) respectively. Propensity score matching of the controls resulted in equalised total costs for the years 2013 through 2015 for clients (n = 50, EUR 21,516 (14,513–28,518)) and controls (n = 250, EUR 21,725 (18,214–25,234)). The total number of integration assistance receivers in the district was lower than the average for the federal state. The number of consultations at the outpatient clinic steadily increased from 146 in 2013 to 1,090 in 2015. Counselling in the outreach clinic might help reduce the placement of clients into integration assistance, including supported housing, and slow the expected cost trend. However, counselling failed to lower total costs in the integration assistance service, possibly due to the selection of more severe cases

Probing the mass function of halo dark matter via microlensing

The simplest interpretation of the microlensing events observed towards the Large Magellanic Clouds is that approximately half of the mass of the Milky Way halo is in the form of MAssive Compact Halo Objects with $M \sim 0.5 M_{\odot}$. It is not possible, due to limits from star counts and chemical abundance arguments, for faint stars or white dwarves to comprise such a large fraction of the halo mass. This leads to the consideration of more exotic lens candidates, such as primordial black holes, or alternative lens locations. If the lenses are located in the halo of the Milky Way, then constraining their mass function will shed light on their nature. Using the current microlensing data we find, for four halo models, the best fit parameters for delta-function, primordial black hole and various power law mass functions. The best fit primordial black hole mass functions, despite having significant finite width, have likelihoods which are similar to, and for one particular halo model greater than, those of the best fit delta functions . We then use Monte Carlo simulations to investigate the number of microlensing events necessary to determine whether the MACHO mass function has significant finite width. If the correct halo model is known, then $\sim$ 500 microlensing events will be sufficient, and will also allow determination of the mass function parameters to $\sim 5$.Comment: 28 pages including 14 figures, version to appear in ApJ, minor changes to discussio

Three dimensional in vitro models of cancer: Bioprinting multilineage glioblastoma models

International audienceThree dimensional (3D) bioprinting of multiple cell types within optimised extracellular matrices has the potential to more closely model the 3D environment of human physiology and disease than current alternatives. In this study, we used a multi-nozzle extrusion bioprinter to establish models of glioblastoma made up of cancer and stromal cells printed within matrices comprised of alginate modified with RGDS cell adhesion peptides, hyaluronic acid and collagen-1. Methods were developed using U87MG glioblastoma cells and MM6 monocyte/macrophages, whilst more disease relevant constructs contained glioblastoma stem cells (GSCs), co-printed with glioma associated stromal cells (GASCs) and microglia. Printing parameters were optimised to promote cell-cell interaction, avoiding the 'caging in' of cells due to overly dense cross-linking. Such printing had a negligible effect on cell viability, and cells retained robust metabolic activity and proliferation. Alginate gels allowed the rapid recovery of printed cell protein and RNA, and fluorescent reporters provided analysis of protein kinase activation at the single cell level within printed constructs. GSCs showed more resistance to chemotherapeutic drugs in 3D printed tumour constructs compared to 2D monolayer cultures, reflecting the clinical situation. In summary, a novel 3D bioprinting strategy is developed which allows control over the spatial organisation of tumour constructs for pre-clinical drug sensitivity testing and studies of the tumour microenvironment

Cosmological Creation of D-branes and anti-D-branes

We argue that the early universe may be described by an initial state of space-filling branes and anti-branes. At high temperature this system is stable. At low temperature tachyons appear and lead to a phase transition, dynamics, and the creation of D-branes. These branes are cosmologically produced in a generic fashion by the Kibble mechanism. From an entropic point of view, the formation of lower dimensional branes is preferred and $D3$ brane-worlds are exponentially more likely to form than higher dimensional branes. Virtually any brane configuration can be created from such phase transitions by adjusting the tachyon profile. A lower bound on the number defects produced is: one D-brane per Hubble volume.Comment: 30 pages, 5 eps figures; v2 more references added; v3 section 4 slightly improve

Targeted genetic testing for familial hypercholesterolaemia using next generation sequencing:a population-based study

Background&lt;p&gt;&lt;/p&gt; Familial hypercholesterolaemia (FH) is a common Mendelian condition which, untreated, results in premature coronary heart disease. An estimated 88% of FH cases are undiagnosed in the UK. We previously validated a method for FH mutation detection in a lipid clinic population using next generation sequencing (NGS), but this did not address the challenge of identifying index cases in primary care where most undiagnosed patients receive healthcare. Here, we evaluate the targeted use of NGS as a potential route to diagnosis of FH in a primary care population subset selected for hypercholesterolaemia.&lt;p&gt;&lt;/p&gt; Methods&lt;p&gt;&lt;/p&gt; We used microfluidics-based PCR amplification coupled with NGS and multiplex ligation-dependent probe amplification (MLPA) to detect mutations in LDLR, APOB and PCSK9 in three phenotypic groups within the Generation Scotland: Scottish Family Health Study including 193 individuals with high total cholesterol, 232 with moderately high total cholesterol despite cholesterol-lowering therapy, and 192 normocholesterolaemic controls.&lt;p&gt;&lt;/p&gt; Results&lt;p&gt;&lt;/p&gt; Pathogenic mutations were found in 2.1% of hypercholesterolaemic individuals, in 2.2% of subjects on cholesterol-lowering therapy and in 42% of their available first-degree relatives. In addition, variants of uncertain clinical significance (VUCS) were detected in 1.4% of the hypercholesterolaemic and cholesterol-lowering therapy groups. No pathogenic variants or VUCS were detected in controls.&lt;p&gt;&lt;/p&gt; Conclusions&lt;p&gt;&lt;/p&gt; We demonstrated that population-based genetic testing using these protocols is able to deliver definitive molecular diagnoses of FH in individuals with high cholesterol or on cholesterol-lowering therapy. The lower cost and labour associated with NGS-based testing may increase the attractiveness of a population-based approach to FH detection compared to genetic testing with conventional sequencing. This could provide one route to increasing the present low percentage of FH cases with a genetic diagnosis

U-Limb: A multi-modal, multi-center database on arm motion control in healthy and post-stroke conditions

BACKGROUND: Shedding light on the neuroscientific mechanisms of human upper limb motor control, in both healthy and disease conditions (e.g., after a stroke), can help to devise effective tools for a quantitative evaluation of the impaired conditions, and to properly inform the rehabilitative process. Furthermore, the design and control of mechatronic devices can also benefit from such neuroscientific outcomes, with important implications for assistive and rehabilitation robotics and advanced human-machine interaction. To reach these goals, we believe that an exhaustive data collection on human behavior is a mandatory step. For this reason, we release U-Limb, a large, multi-modal, multi-center data collection on human upper limb movements, with the aim of fostering trans-disciplinary cross-fertilization. CONTRIBUTION: This collection of signals consists of data from 91 able-bodied and 65 post-stroke participants and is organized at 3 levels: (i) upper limb daily living activities, during which kinematic and physiological signals (electromyography, electro-encephalography, and electrocardiography) were recorded; (ii) force-kinematic behavior during precise manipulation tasks with a haptic device; and (iii) brain activity during hand control using functional magnetic resonance imaging

The Threat of Capital Drain: A Rationale for Public Banks?

This paper yields a rationale for why subsidized public banks may be desirable from a regional perspective in a financially integrated economy. We present a model with credit rationing and heterogeneous regions in which public banks prevent a capital drain from poorer to richer regions by subsidizing local depositors, for example, through a public guarantee. Under some conditions, cooperative banks can perform the same function without any subsidization; however, they may be crowded out by public banks. We also discuss the impact of the political structure on the emergence of public banks in a political-economy setting and the role of interregional mobility