Guided mode resonance sensor for the parallel detection of multiple protein biomarkers in human urine with high sensitivity

The rising cost of global healthcare provision and new approaches to managing disease are driving the development of low-cost biosensing modalities, such as label-free photonic methods based on dielectric resonances. Here, we use the combined sensing and imaging capability of a guided mode resonance (GMR) sensor to detect multiple biomarkers (troponin, procalcitonin and C-Reactive Protein) in parallel in undiluted urine samples. A key requirement of such a biosensor is the simple and direct functionalization with suitable antibodies to ensure the disease-specific detection of protein biomarkers. Here, antibodies were immobilized using a succinimidyl-[(N-maleimidopropionamido)-hexaethyleneglycol] ester (SM(PEG)6) spacer. The polyethylene glycol (PEG) chemistry enables low detection limits of 10 pg mL-1 or better for all protein biomarkers, while minimizing non-specific binding compared to more commonly used strategies such as (3-Aminopropyl)triethoxysilane (APTES) or dextran. Our approach supports the vision of a simple yet highly sensitive diagnostic platform that could be used for pre-screening patients for a wide range of diseases at point-of-care, thereby relieving the pressure on overstretched healthcare services

Site-selectively grown SnO2 NWs networks on micromembranes for efficient ammonia sensing in humid conditions

SnO2 NWs networks on heated micromembranes have been characterized as ammonia sensors. The approach allows achieving reproducible growth and stable and long-lasting ammonia sensors with site-specific grown SnO2 NWs. The devices have been tested both in dry and humid conditions showing response time down to two minutes. Sensors have been tested up to 1 month, only presenting variation of the base resistance with full retention of the response towards the gaseous analytes. Different concurrent sensing mechanisms have been identified relating the determined sensing kinetics with previous theoretical calculations. Specifically, oxygen dissociation seems to play a key role in the overall ammonia sensing sequence. In humid conditions, moisture reduces the response to ammonia but also lowers the activation energy of the reaction process

EST-derived SSR markers used as anchor loci for the construction of a consensus linkage map in ryegrass (Lolium spp.)

BACKGROUND: Genetic markers and linkage mapping are basic prerequisites for marker-assisted selection and map-based cloning. In the case of the key grassland species Lolium spp., numerous mapping populations have been developed and characterised for various traits. Although some genetic linkage maps of these populations have been aligned with each other using publicly available DNA markers, the number of common markers among genetic maps is still low, limiting the ability to compare candidate gene and QTL locations across germplasm. RESULTS: A set of 204 expressed sequence tag (EST)-derived simple sequence repeat (SSR) markers has been assigned to map positions using eight different ryegrass mapping populations. Marker properties of a subset of 64 EST-SSRs were assessed in six to eight individuals of each mapping population and revealed 83% of the markers to be polymorphic in at least one population and an average number of alleles of 4.88. EST-SSR markers polymorphic in multiple populations served as anchor markers and allowed the construction of the first comprehensive consensus map for ryegrass. The integrated map was complemented with 97 SSRs from previously published linkage maps and finally contained 284 EST-derived and genomic SSR markers. The total map length was 742 centiMorgan (cM), ranging for individual chromosomes from 70 cM of linkage group (LG) 6 to 171 cM of LG 2. CONCLUSIONS: The consensus linkage map for ryegrass based on eight mapping populations and constructed using a large set of publicly available Lolium EST-SSRs mapped for the first time together with previously mapped SSR markers will allow for consolidating existing mapping and QTL information in ryegrass. Map and markers presented here will prove to be an asset in the development for both molecular breeding of ryegrass as well as comparative genetics and genomics within grass species

Bio-inspired polydopamine layer as a versatile functionalisation protocol for silicon-based photonic biosensors

Photonic biosensors have made major advances in recent years, achieving very high sensitivity, and progressing towards point-of-care deployment. By using photonic resonances, sensors can be label-free, which is particularly attractive for a low-cost technological realisation. A key remaining issue is the biological interface and the efficient and reliable immobilisation of binder molecules such as antibodies; many protocols are currently in use that have led to widely varying sensor performance. Here, we study a very simple and robust surface functionalisation protocol for silicon photonics, which is based on polydopamine, and we demonstrate both its simplicity and its high performance. The use of polydopamine (PDA) is inspired by molluscs, especially mussels, that employ dopamine to adhere to virtually any surface, especially in an aqueous environment. We studied the versatility of the PDA protocol by showing compatibility with 5 different disease biomarkers (Immunoglobulin (IgG), C-reactive protein (CRP), Tumour Necrosis factor-α (TNF-α), Interleukin-6 (IL-6), Matrix metalloproteinase (MMP-9) and show that the protocol is resistant to hydrolysis during incubation; the loss of functionality due to hydrolysis is a major issue for many of the functionalisation protocols commonly used for silicon-based sensors. The study using guided mode resonance-based sensors highlights the wide dynamic range of the protocol (0.01 ng/mL to 1 μg/mL), using IgG, CRP and MMP-9 protein biomarkers as exemplars. In addition, we show that the surface chemistry allows performing measurements in 10% human serum with a sensitivity as low as 10 ng/mL for IgG. We suggest that adopting this protocol will make it easier for researchers to achieve biofunctionalisation and that the biosensor community will be able to achieve more consistent results

Extended Kalman Filtering Projection Method to Reduce the 3σ Noise Value of Optical Biosensors

Optical biosensors have experienced a rapid growth over the past decade because of their high sensitivity and the fact that they are label-free. Many optical biosensors rely on tracking the change in a resonance signal or an interference pattern caused by the change in refractive index that occurs upon binding to a target biomarker. The most commonly used method for tracking such a signal is based on fitting the data with an appropriate mathematical function, such as a harmonic function or a Fano, Gaussian, or Lorentz function. However, these functions have limited fitting efficiency because of the deformation of data from noise. Here, we introduce an extended Kalman filter projection (EKFP) method to address the problem of resonance tracking and demonstrate that it improves the tolerance to noise, reduces the 3σ noise value, and lowers the limit of detection (LOD). We utilize the method to process the data of experiments for detecting the binding of C-reactive protein in a urine matrix with a chirped guided mode resonance sensor and are able to improve the LOD from 10 to 1 pg/mL. Our method reduces the 3σ noise value of this measurement compared to a simple Fano fit from 1.303 to 0.015 pixels. These results demonstrate the significant advantage of the EKFP method to resolving noisy data of optical biosensors

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

Hepatitis C virus cell-cell transmission and resistance to direct-acting antiviral agents

Hepatitis C virus (HCV) is transmitted between hepatocytes via classical cell entry but also uses direct cell-cell transfer to infect neighboring hepatocytes. Viral cell-cell transmission has been shown to play an important role in viral persistence allowing evasion from neutralizing antibodies. In contrast, the role of HCV cell-cell transmission for antiviral resistance is unknown. Aiming to address this question we investigated the phenotype of HCV strains exhibiting resistance to direct-acting antivirals (DAAs) in state-of-the-art model systems for cell-cell transmission and spread. Using HCV genotype 2 as a model virus, we show that cell-cell transmission is the main route of viral spread of DAA-resistant HCV. Cell-cell transmission of DAA-resistant viruses results in viral persistence and thus hampers viral eradication. We also show that blocking cell-cell transmission using host-targeting entry inhibitors (HTEIs) was highly effective in inhibiting viral dissemination of resistant genotype 2 viruses. Combining HTEIs with DAAs prevented antiviral resistance and led to rapid elimination of the virus in cell culture model. In conclusion, our work provides evidence that cell-cell transmission plays an important role in dissemination and maintenance of resistant variants in cell culture models. Blocking virus cell-cell transmission prevents emergence of drug resistance in persistent viral infection including resistance to HCV DAAs

Деякі проблеми використання тимчасово зайнятих земель

<div><p>Glucocorticoid induced-leucine zipper (GILZ) has been shown to be induced in cells by different stimuli such as glucocorticoids, IL-10 or deprivation of IL-2. GILZ has anti-inflammatory properties and may be involved in signalling modulating apoptosis. Herein we demonstrate that wildtype <em>Yersinia enterocolitica</em> which carry the pYV plasmid upregulated GILZ mRNA levels and protein expression in epithelial cells. Infection of HeLa cells with different <em>Yersinia</em> mutant strains revealed that the protease activity of YopT, which cleaves the membrane-bound form of Rho GTPases was sufficient to induce GILZ expression. Similarly, <em>Clostridium difficile</em> toxin B, another bacterial inhibitor of Rho GTPases induced GILZ expression. YopT and toxin B both increased transcriptional activity of the GILZ promoter in HeLa cells. GILZ expression could not be linked to the inactivation of an individual Rho GTPase by these toxins. However, forced expression of RhoA and RhoB decreased basal <em>GILZ</em> promoter activity. Furthermore, MAPK activation proved necessary for profound GILZ induction by toxin B. Promoter studies and gel shift analyses defined binding of upstream stimulatory factor (USF) 1 and 2 to a canonical c-Myc binding site (E-box) in the <em>GILZ</em> promoter as a crucial step of its trans-activation. In addition we could show that USF-1 and USF-2 are essential for basal as well as toxin B induced GILZ expression. These findings define a novel way of <em>GILZ</em> promoter trans-activation mediated by bacterial toxins and differentiate it from those mediated by dexamethasone or deprivation of IL-2.</p> </div

Locally grown SnO2 NWs as low power ammonia sensor

Localized growth of SnO2 nanowires on top of CMOS compatible micromembranes that incorporate a buried heater and prepatterned interdigitated electrodes has been achieved that presents the advantage that it allows to easily and directly integrate the advantageous properties of quasi-one dimensional structures in an advanced electronic device by a Vapor Liquid Solid (VLS) mechanism. A NWs based sensor of this type is characterized as a low power gas sensor towards NH3 at different temperatures. Stable and reproducible response is obtained, that allows detecting concentrations below the time-weighted average exposure limit for 8 h

Yarrowia lipolytica growth under increased air pressure: influence on enzymes production

Improvement of microbial cell cultures oxygenation can be achieved by the increase of total air pressure, which increases oxygen solubility in the medium. In this work, a pressurized bioreactor was used for Yarrowia lipolytica batch cultivation under increased air pressure from 1 to 6 bar. Cell growth was strongly enhanced by the pressure rise. Fivefold and 3.4-fold increases in the biomass production and in specific growth rate, respectively, were observed under 6 bar. The increase of oxygen availability caused the induction of the antioxidant enzyme superoxide dismutase, which indicates that the defensive mechanisms of the cells against oxidative stress were effective and cells could cope with increased pressure. The pregrowth of Y. lipolytica under increased pressure conditions did not affect the lipase production ability of the cells. Moreover, the extracellular lipase activity increased 96% using a 5-bar air pressure instead of air at 1- bar pressure during the enzyme production phase. Thus, air pressure increase in bioreactors is an effective mean of cell mass and enzyme productivity enhancement in bioprocess based in Y. lipolytica cultures