Challenges with Point-Of-Care Tests (POCT) for Celiac Disease

Current screening test for celiac disease involves blood test in centralized pathology laboratories, typically performing enzyme-linked immune-sorbent assays (ELISA) to detect specific celiac disease antibodies. Most of the current available celiac disease antibody tests detect anti-gliadin (AGA), anti-endomysial (EMA), anti-transglutaminase (tTG), or deamidated gluten peptide (DGP) antibodies from serum or whole blood samples. It requires blood collection from untreated celiac patients, which is often invasive and inconvenient. There is a rapid growth in demand for noninvasive celiac tests for the early and fast diagnosis of celiac disease to help potential celiac patients obtain results and take corresponding actions. Over the last decade, several point-of-care tests (POCT) have been introduced to the market, but these tests have not been widely accepted by clinicians. Moreover, the 2009 NICE guideline CG 86 recommended that self-tests and/or POCT for celiac disease should not be used as a substitute for laboratory-based tests. Here, we provide a background on the evolution of POCT for celiac disease. We discuss general principle of operation for the known commercial kits as well as the use of various antigens and antibodies in different tests developed over the years. Finally, we discuss challenges for future research directions in celiac disease POCTs

Facile Self-Assembly of Quantum Plasmonic Circuit Components

Efficient coupling between solid state quantum emitters and plasmonic waveguides is important for the realization of integrated circuits for quantum information, communication and sensing. However, realization of plasmonic circuits is still scarce, particularly due to challenges associated with accurate positioning of quantum emitters near plasmonic resonators. Current pathways for the construction of plasmonic circuits involve cumbersome and costly methods such as scanning atomic force microscopy or mechanical manipulation, where individual elements are physically relocated using the scanning tip. Here, we introduce a simple, fast and cost effective chemical self-assembly method for the attachment of two primary components of a practical plasmonic circuit: a single photon emitter and a waveguide. Our method enables coupling of nanodiamonds with a single quantum emitter (the nitrogen-vacancy (NV) center) onto the terminal of a silver nanowire, by simply varying the concentration of ascorbic acid (AA) in a reaction solution. The AA concentration is used to control the extent of agglomeration, and can be optimised so as to cause preferential, selective activation of the tips of the nanowires. The nanowire-nanodiamond structures show efficient plasmonic coupling of fluorescence emission from single NV centers into surface plasmon polariton (SPP) modes, evidenced by a more than two-fold reduction in fluorescence lifetime and an increase in fluorescence intensity.Comment: Published in Advanced Materials on 2 June 201

Ultra-bright emission from hexagonal boron nitride defects as a new platform for bio-imaging and bio-labelling

Bio-imaging requires robust ultra-bright probes without causing any toxicity to the cellular environment, maintain their stability and are chemically inert. In this work we present hexagonal boron nitride (hBN) nanoflakes which exhibit narrowband ultra-bright single photon emitters1. The emitters are optically stable at room temperature and under ambient environment. hBN has also been noted to be noncytotoxic and seen significant advances in functionalization with biomolecules2,3. We further demonstrate two methods of engineering this new range of extremely robust multicolour emitters across the visible and near infrared spectral ranges for large scale sensing and biolabeling applications

Characterization of Cyclin E Expression in Multiple Myeloma and Its Functional Role in Seliciclib-Induced Apoptotic Cell Death

Multiple Myeloma (MM) is a lymphatic neoplasm characterized by clonal proliferation of malignant plasma cell that eventually develops resistance to chemotherapy. Drug resistance, differentiation block and increased survival of the MM tumor cells result from high genomic instability. Chromosomal translocations, the most common genomic alterations in MM, lead to dysregulation of cyclin D, a regulatory protein that governs the activation of key cell cycle regulator – cyclin dependent kinase (CDK). Genomic instability was reported to be affected by over expression of another CDK regulator - cyclin E (CCNE). This occurs early in tumorigenesis in various lymphatic malignancies including CLL, NHL and HL. We therefore sought to investigate the role of cyclin E in MM. CCNE1 expression was found to be heterogeneous in various MM cell lines (hMMCLs). Incubation of hMMCLs with seliciclib, a selective CDK-inhibitor, results in apoptosis which is accompanied by down regulation of MCL1 and p27. Ectopic over expression of CCNE1 resulted in reduced sensitivity of the MM tumor cells in comparison to the paternal cell line, whereas CCNE1 silencing with siRNA increased the cell sensitivity to seliciclib. Adhesion to FN of hMMCLs was prevented by seliciclib, eliminating adhesion–mediated drug resistance of MM cells. Combination of seliciclib with flavopiridol effectively reduced CCNE1 and CCND1 protein levels, increased subG1 apoptotic fraction and promoted MM cell death in BMSCs co-culture conditions, therefore over-coming stroma-mediated protection. We suggest that seliciclib may be considered as essential component of modern anti MM drug combination therapy

Poly(methacrylic acid) hydrogel capsules as a platform for biomedical applications

© 2012 Dr. Olga ShimoniThe design and assembly of biocompatible nanoengineered carriers is of interest due to their potential applications in biotechnology as tools for catalysis and sensing, in biomedicine as systems for drug delivery, in diagnostics, and in vivo imaging. The layer-by-layer (LbL) technology is a prominent technique to design carrier systems for biomedical applications. In recent years, poly(methacrylic acid) hydrogel capsules (PMA HCs), based on disulfide-stabilized poly(methacrylic acid), have been fabricated from the LbL technique, and thoroughly studied to gain control over their stability, degradability and cargo release. These capsules are obtained by the sequential deposition of thiolated poly(methacrylic acid) (PMASH) and poly(N-vinylpyrrolidone) (PVPON) onto silica particles via hydrogen bonding. Upon controlled crosslinking and removal of the silica template, PVPON is released at physiological conditions due to the disrupted hydrogen bonding between PMASH and PVPON, which results in single-component PMA hydrogel capsules. This work provides an insight into designing a novel architecture and biofunctionalization of PMA HCs for enhanced and targeted drug delivery. Specifically, this research describes novel hydrogel capsule architectures, namely subcompartmentalized hydrogel capsules (SHCs), which are designed for potential applications in drug delivery and microencapsulated biocatalysis. Examples of SHCs with tens of subcompartments are demonstrated with their successful drug/cargo loading, as well as selective degradation of the SHC carrier and/or sub-units in response to multiple chemical stimuli. To develop a facile surface functionalization approach of the PMA hydrogel capsules, retention of PVPON was employed through modification of the polymer to obtain a bifunctional polymeric linker. The antibody-functionalized PMA/PVPON HCs demonstrate significantly enhanced cellular binding and internalization to specific cells, suggesting these capsules can specifically interact with cells through antibody/antigen recognition. To understand the impact of aspect ratio on cellular function, PMA HCs were prepared with various aspect ratios. Careful control over aspect ratio of the silica rods provided the ability to control the aspect ratio of the PMA HCs. Upon incubation of these capsules with living cells, varied behavior was observed, suggesting different mechanisms for their interactions with cells

Shape-Dependent Cellular Processing of Polyelectrolyte Capsules

Particle shape is emerging as a key design parameter for tailoring the interactions between particles and cells. Herein, we report the preparation of rod-shaped layer-by-layer (LbL)-assembled polymer hydrogel capsules with tunable aspect ratios (ARs). By templating spherical and rodlike silica particles, disulfide-stabilized poly(methacrylic acid) hydrogel capsules (PMA HCs) with different ARs (from 1 to 4) are generated. The influence of capsule AR on cellular internalization and intracellular fate was quantitatively investigated by flow cytometry, imaging flow cytometry, and fluorescence deconvolution microscopy. These experiments reveal that the cellular internalization kinetics of PMA HCs are dependent on the AR, with spherical capsules being internalized more rapidly and to a greater extent compared with rod-shaped capsules. In contrast, the capsules with different ARs are colocalized with the lysosomal marker LAMP1, suggesting that the lysosomal compartmentalization is independent of shape for these soft polymer capsules

CMV Seropositive Status Increases Heparanase SNPs Regulatory Activity, Risk of Acute GVHD and Yield of CD34+ Cell Mobilization

Heparanase is an endo-β-glucuronidase that is best known for its pro-cancerous effects but is also implicated in the pathogenesis of various viruses. Activation of heparanase is a common strategy to increase viral spread and trigger the subsequent inflammatory cascade. Using a Single Nucleotide Polymorphisms (SNP)-associated approach we identified enhancer and insulator regions that regulate HPSE expression. Although a role for heparanase in viral infection has been noticed, the impact of HPSE functional SNPs has not been determined. We investigated the effect of cytomegalovirus (CMV) serostatus on the involvement of HPSE enhancer and insulator functional SNPs in the risk of acute graft versus host disease (GVHD) and granulocyte-colony stimulating factor related CD34+ mobilization. A significant correlation between the C alleles of insulator rs4364254 and rs4426765 and CMV seropositivity was found in healthy donors and patients with hematological malignancies. The risk of developing acute GVHD after hematopoietic stem cell transplantation was identified only in CMV-seropositive patients. A significant correlation between the enhancer rs4693608 and insulator rs28649799 and CD34+ cell mobilization was demonstrated in the CMV-seropositive donors. It is thus conceivable that latent CMV infection modulates heparanase regulatory regions and enhances the effect of functional SNPs on heparanase function in normal and pathological processes

The HPSE Gene Insulator—A Novel Regulatory Element That Affects Heparanase Expression, Stem Cell Mobilization, and the Risk of Acute Graft versus Host Disease

The HPSE gene encodes heparanase (HPSE), a key player in cancer, inflammation, and autoimmunity. We have previously identified a strong HPSE gene enhancer involved in self-regulation of heparanase by negative feedback exerted in a functional rs4693608 single-nucleotide polymorphism (SNP) dependent manner. In the present study, we analyzed the HPSE gene insulator region, located in intron 9 and containing rs4426765, rs28649799, and rs4364254 SNPs. Our results indicate that this region exhibits HPSE regulatory activity. SNP substitutions lead to modulation of a unique DNA-protein complex that affects insulator activity. Analysis of interactions between enhancer and insulator SNPs revealed that rs4693608 has a major effect on HPSE expression and the risk of post-transplantation acute graft versus host disease (GVHD). The C alleles of insulator SNPs rs4364254 and rs4426765 modify the activity of the HPSE enhancer, resulting in altered HPSE expression and increased risk of acute GVHD. Moreover, rs4426765 correlated with HPSE expression in activated mononuclear cells, as well as with CD3 levels and lymphocyte counts following G-CSF mobilization. rs4363084 and rs28649799 were found to be associated with CD34+ levels. Our study provides new insight into the mechanism of HPSE gene regulation and its impact on normal and pathological processes in the hematopoietic system