Tough Adhesive Hydrogel for Intraoral Adhesion and Drug Delivery.

Oral lichen planus (OLP) and recurrent aphthous stomatitis (RAS) are common chronic inflammatory conditions, manifesting as painful oral lesions that negatively affect patients' quality of life. Current treatment approaches are mainly palliative and often ineffective due to inadequate contact time of the therapeutic agent with the lesions. Here, we developed the Dental Tough Adhesive (DenTAl), a bioinspired adhesive patch with robust mechanical properties, capable of strong adhesion against diverse wet and dynamically moving intraoral tissues, and extended drug delivery of clobetasol-17-propionate, a first-line drug for treating OLP and RAS. DenTAl was found to have superior physical and adhesive properties compared to existing oral technologies, with ~2 to 100× adhesion to porcine keratinized gingiva and ~3 to 15× stretchability. Clobetasol-17-propionate incorporated into the DenTAl was released in a tunable sustained manner for at least 3 wk and demonstrated immunomodulatory capabilities in vitro, evidenced by reductions in several cytokines, including TNF-α, IL-6, IL-10, MCP-5, MIP-2, and TIMP-1. Our findings suggest that DenTAl may be a promising device for intraoral delivery of small-molecule drugs applicable to the management of painful oral lesions associated with chronic inflammatory conditions.

Safety of the use of gold nanoparticles conjugated with proinsulin peptide and administered by hollow microneedles as an immunotherapy in type 1 diabetes.

Antigen-specific immunotherapy is an immunomodulatory strategy for autoimmune diseases, such as type 1 diabetes, in which patients are treated with autoantigens to promote immune tolerance, stop autoimmune ß-cell destruction and prevent permanent dependence on exogenous insulin. In this study, human proinsulin peptide C19-A3 (known for its positive safety profile) was conjugated to ultrasmall gold nanoparticles (GNPs), an attractive drug delivery platform due to the potential anti-inflammatory properties of gold. We hypothesised that microneedle intradermal delivery of C19-A3 GNP may improve peptide pharmacokinetics and induce tolerogenic immunomodulation and proceeded to evaluate its safety and feasibility in a first-in-human trial. Allowing for the limitation of the small number of participants, intradermal administration of C19-A3 GNP appears safe and well tolerated in participants with type 1 diabetes. The associated prolonged skin retention of C19-A3 GNP after intradermal administration offers a number of possibilities to enhance its tolerogenic potential, which should be explored in future studies.

Proteomic analysis reveals greater abundance of complement and inflammatory proteins in subcutaneous adipose tissue from postpartum cows treated with sodium salicylate.

This study aimed to investigate sodium salicylate (SS) treatment effects on the proteome of adipose tissue (AT) in postpartum cows. Twenty Holstein cows were assigned to control (CON, n = 10) or SS (n = 10) provided via drinking water (2.3 g/L) during the first 7 d of lactation. Subcutaneous AT was collected on d 7 of treatment and label-free quantitative shotgun proteomics and immunoblotting were analyzed in a subset of 5 AT per group. Eighty out of 1422 proteins (5.6%) were differentially abundant between CON and SS [fold change ±1.5, P < 0.05]. Top canonical pathways differing between CON and SS (Ingenuity) were complement system, interleukin-10 signaling, and acute phase response signaling. The abundances of complement C1r, C1qC, C1qB and C6 were greater in SS than CON. Regarding IL-10 signaling, the abundances of BLVRB, STAT3, and lipopolysaccharide binding protein (LBP) were greater in SS AT compared to CON. Immunoblots revealed increased abundance of paraoxanase-1 and tumor necrosis factor-alpha, as well as a tendency for greater abundance of cluster differentiation 172a in SS AT, which may indicate of increased macrophage infiltration. SS treatment postpartum likely promotes inflammatory signaling in AT of dairy cows, perhaps due to immune cell recruitment. SIGNIFICANCE: This work demonstrates that treating early lactating cows with sodium salicylate, an anti-inflammatory agent that has been shown to have metabolic effects and increase milk production in dairy cows, affects the proteome of subcutaneous adipose tissue in early lactating dairy cows. Unexpectedly, sodium salicylate treatment enriched inflammatory pathways of the complement system, cytokine signaling, and acute phase response, as revealed by proteomic analysis of subcutaneous adipose tissues from cows at 7 d postpartum. These findings imply that SS treatment during the first 7 d of lactation likely promotes inflammatory signaling in AT of the dairy cow, perhaps due to immune cell recruitment. Tissue-specific impacts of systemic sodium salicylate requires further scrutiny.

Mono(2-ethylhexyl) phthalate (MEHP) induces transcriptomic alterations in oocytes and their derived blastocysts.

Mono(2-ethylhexyl) phthalate (MEHP), the main di(2-ethylhexyl) phthalate (DEHP) metabolite, is a known reproductive toxicant. Residual levels of 20 nM MEHP have been found in follicular fluid aspirated from IVF-treated women and DEHP-treated animals. The current study examined whether these residual MEHP levels have any effect on the follicle-enclosed oocyte or developing embryo. Bovine oocytes were matured with or without 20 nM MEHP for 22 h. Microarray analysis was performed for both mature oocytes and 7-day blastocysts. A proteomic analysis was performed on mature oocytes (n = 200/group) to reveal a possible direct effect on the oocyte proteomic profile. Transcriptome analysis revealed MEHP-induced alterations in the expression of 456 and 290 genes in oocytes and blastocysts, respectively. The differentially expressed genes are known to be involved in various biological pathways, such as transcription process, cytoskeleton regulation and metabolic pathway. Among these, the expression of 9 genes was impaired in both oocytes exposed to MEHP (i.e., direct effect) and blastocysts developed from those oocytes (i.e., carryover effect). In addition, 191 proteins were found to be affected by MEHP in mature oocytes (Data are available via ProteomeXchange with identifier PXD012092). The study explores, for the first time, the risk associated with exposing oocytes to low concentration (i.e., environmentally relevant concentration) of MEHP to the maternal transcripts. Although it was the oocytes that were exposed to MEHP, alterations carried over to the blastocyst stage, following embryonic genome activation, implying that these embryos are of low quality.

Conjugation of a peptide autoantigen to gold nanoparticles for intradermally administered antigen specific immunotherapy.

Antigen specific immunotherapy aims to tolerise patients to specific autoantigens that are responsible for the pathology of an autoimmune disease. Immune tolerance is generated in conditions where the immune response is suppressed and thus gold nanoparticles (AuNPs) are an attractive drug delivery platform due to their anti-inflammatory effects and their potential to facilitate temporal and spatial delivery of a peptide autoantigen in conjunction with pro-tolerogenic elements. In this study we have covalently attached an autoantigen, currently under clinical evaluation for the treatment of type 1 diabetes (PIC19-A3 peptide), to AuNPs to create nanoscale (<5 nm), negatively charged (-40 to -60 mV) AuNP-peptide complexes for immunotherapy. We also employ a clinically approved microneedle delivery system, MicronJet600, to facilitate minimally-invasive intradermal delivery of the nanoparticle constructs to target skin-resident antigen presenting cells, which are known to be apposite target cells for immunotherapy. The AuNP-peptide complexes remain physically stable upon extrusion through microneedles and when delivered into ex vivo human skin they are able to diffuse rapidly and widely throughout the dermis (their site of deposition) and, perhaps more surprisingly, the overlying epidermal layer. Intracellular uptake was extensive, with Langerhans cells proving to be the most efficient cells at internalising the AuNP-peptide complex (94% of the local population within the treated region of skin). In vitro studies showed that uptake of the AuNP-peptide complexes by dendritic cells reduced the capacity of these cells to activate naïve T cells. This indicator of biological functionality encourages further development of the AuNP-peptide formulation, which is now being evaluated in clinical trials.

Proteome dataset of subcutaneous adipose tissue obtained from late pregnant dairy cows during summer heat stress and winter seasons.

Adipose tissue has a central role in the regulation of metabolism in dairy cows, and many proteins expressed in this tissue are involved in metabolic responses to stress (Peinado et al., 2012) [1]. Environmental heat stress is one of the main stressors limiting production in dairy cattle (Fuquay, 1981; West, 2003) [2], [3], and there is a complex interaction between heat stress and the transition period from late pregnancy to onset of lactation, which is manifested in heat-stressed late-gestation cows (Tao and Dahl, 2013) [4]. We recently defined the proteome of adipose tissue in peripartum dairy cows, identifying 586 proteins of which 18.9% were differentially abundant in insulin-resistant compared to insulin-sensitive adipose tissue (Zachut, 2015) [5]. That study showed that proteomic techniques constitute a valuable tool for identifying novel biomarkers in adipose tissue that are related to metabolic adaptation to stress in dairy cows. The objective of the present work was to examine the adipose tissue proteome under thermo-neutral or seasonal heat stress conditions in late pregnant dairy cows. We have collected subcutaneous adipose tissue biopsies from 10 late pregnant dairy cows during summer heat stress and from 8 late pregnant dairy cows during winter season, and identified and quantified 1495 proteins in the adipose tissues. This dataset of adipose tissue proteome from dairy cows adds novel information on the variety of proteins that are abundant in this tissue during late pregnancy under thermo-neutral as well as heat stress conditions. Differential abundance of 107 (7.1%) proteins was found between summer and winter adipose. These results are discussed in our recent research article (Zachut et al., 2017) [6].

Hydrodynamic gene delivery in human skin using a hollow microneedle device.

Microneedle devices have been proposed as a minimally invasive delivery system for the intradermal administration of nucleic acids, both plasmid DNA (pDNA) and siRNA, to treat localised disease or provide vaccination. Different microneedle types and application methods have been investigated in the laboratory, but limited and irreproducible levels of gene expression have proven to be significant challenges to pre-clinical to clinical progression. This study is the first to explore the potential of a hollow microneedle device for the delivery and subsequent expression of pDNA in human skin. The regulatory approved MicronJet600® (MicronJet hereafter) device was used to deliver reporter plasmids (pCMVß and pEGFP-N1) into viable excised human skin. Exogenous gene expression was subsequently detected at multiple locations that were distant from the injection site but within the confines of the bleb created by the intradermal bolus. The observed levels of gene expression in the tissue are at least comparable to that achieved by the most invasive microneedle application methods e.g. lateral application of a microneedle. Gene expression was predominantly located in the epidermis, although also evident in the papillary dermis. Optical coherence tomography permitted real time visualisation of the sub-surface skin architecture and, unlike a conventional intradermal injection, MicronJet administration of a 50µL bolus appears to create multiple superficial microdisruptions in the papillary dermis and epidermis. These were co-localised with expression of the pCMVß reporter plasmid. We have therefore shown, for the first time, that a hollow microneedle device can facilitate efficient and reproducible gene expression of exogenous naked pDNA in human skin using volumes that are considered to be standard for intradermal administration, and postulate a hydrodynamic effect as the mechanism of gene delivery.

Seasonal heat stress affects adipose tissue proteome toward enrichment of the Nrf2-mediated oxidative stress response in late-pregnant dairy cows.

Environmental heat stress and metabolic stress during transition from late gestation to lactation are main factors limiting production in dairy cattle, and there is a complex interaction between them. Many proteins expressed in adipose tissue are involved in metabolic responses to stress. We aimed to investigate the effects of seasonal heat stress on adipose proteome in late-pregnant cows, and to identify biomarkers of heat stress. Late pregnant cows during summer heat stress (S, n=18), or during the winter season (W, n=12) were used. Subcutaneous adipose tissue biopsies sampled 14days prepartum from S (n=10) and W (n=8) were analyzed by intensity-based, label-free, quantitative shotgun proteomics (nano-LC-MS/MS). Plasma concentrations of malondialdehyde and cortisol were higher in S than in W cows. Proteomic analysis revealed that 107/1495 proteins were differentially abundant in S compared to W (P<0.05 and fold change of at least ±1.5). Top canonical pathways in S vs. W adipose were Nrf2-mediated oxidative stress response, acute-phase response, and FXR/RXR and LXR/RXR activation. Novel biomarkers of heat stress in adipose tissue were found. These findings indicate that seasonal heat stress has a unique effect on adipose tissue in late-pregnant cows. SIGNIFICANCE: This work shows that seasonal heat stress increases plasma concentrations of the oxidative stress marker malondialdehyde and cortisol in transition dairy cows. As many proteins expressed in the adipose tissue are involved in metabolic responses to stress, we investigated the effects of heat stress on the proteome of adipose tissue from late-pregnant cows during summer or winter seasons. We demonstrated that heat stress enriches several stress-related pathways, such as the Nrf2-mediated oxidative stress response and the acute-phase response in adipose tissues. Thus, environmental heat stress has a unique effect on adipose tissue in late-pregnant cows, as part of the regulatory adaptations to chronic heat load during the summer season. In addition, this study presents the widest available dataset of adipose tissue proteome in dairy cows, and revealed several novel biomarkers of heat stress in adipose tissue of dairy cows, the use of which awaits further validation.

Influence of network topology on the swelling of polyelectrolyte nanogels.

It is well-known that the swelling behavior of ionic nanogels depends on their cross-link density; however, it is unclear how different topologies should affect the response of the polyelectrolyte network. Here we perform Monte Carlo simulations to obtain the equilibrium properties of ionic nanogels as a function of salt concentration Cs and the fraction f of ionizable groups in a polyelectrolyte network formed by cross-links of functionality z. Our results indicate that the network with cross-links of low connectivity result in nanogel particles with higher swelling ratios. We also confirm a de-swelling effect of salt on nanogel particles.

Gravitational waves from binary supermassive black holes missing in pulsar observations.

Gravitational waves are expected to be radiated by supermassive black hole binaries formed during galaxy mergers. A stochastic superposition of gravitational waves from all such binary systems would modulate the arrival times of pulses from radio pulsars. Using observations of millisecond pulsars obtained with the Parkes radio telescope, we constrained the characteristic amplitude of this background, A(c,yr), to be <1.0 × 10(-15) with 95% confidence. This limit excludes predicted ranges for A(c,yr) from current models with 91 to 99.7% probability. We conclude that binary evolution is either stalled or dramatically accelerated by galactic-center environments and that higher-cadence and shorter-wavelength observations would be more sensitive to gravitational waves.

Gravitational-wave limits from pulsar timing constrain supermassive black hole evolution.

The formation and growth processes of supermassive black holes (SMBHs) are not well constrained. SMBH population models, however, provide specific predictions for the properties of the gravitational-wave background (GWB) from binary SMBHs in merging galaxies throughout the universe. Using observations from the Parkes Pulsar Timing Array, we constrain the fractional GWB energy density (Ω(GW)) with 95% confidence to be Ω(GW)(H0/73 kilometers per second per megaparsec)(2) < 1.3 × 10(-9) (where H0 is the Hubble constant) at a frequency of 2.8 nanohertz, which is approximately a factor of 6 more stringent than previous limits. We compare our limit to models of the SMBH population and find inconsistencies at confidence levels between 46 and 91%. For example, the standard galaxy formation model implemented in the Millennium Simulation Project is inconsistent with our limit with 50% probability.

Ion fluxes through nanopores and transmembrane channels.

We introduce an implicit solvent Molecular Dynamics approach for calculating ionic fluxes through narrow nanopores and transmembrane channels. The method relies on a dual-control-volume grand-canonical molecular dynamics (DCV-GCMD) simulation and the analytical solution for the electrostatic potential inside a cylindrical nanopore recently obtained by Levin [Europhys. Lett. 76, 163 (2006)]. The theory is used to calculate the ionic fluxes through an artificial transmembrane channel which mimics the antibacterial gramicidin A channel. Both current-voltage and current-concentration relations are calculated under various experimental conditions. We show that our results are comparable to the characteristics associated to the gramicidin A pore, especially the existence of two binding sites inside the pore and the observed saturation in the current-concentration profiles.

Novel therapeutic approaches in pulmonary arterial hypertension: focus on tadalafil.

Pulmonary arterial hypertension (PAH) is an enigmatic, often fatal disease of the lung. Excess vasoconstriction and progressive obliteration of the precapillary arterioles combine to reduce the cross-sectional area for blood flow and thus cause chronic elevation in the pulmonary arterial pressures with progressive right heart dysfunction, heart failure and death. In 1995, the FDA approved the first therapy for PAH: epoprostenol, a highly efficacious drug but one that was difficult to use for patients and clinicians alike. Since then, there have been eight additional drugs approved, each offering advantages in terms of convenience over previously available drugs. In 2009, tadalafil (Adcirca®) was approved for PAH. The 405 patients enrolled in the single pivotal trial give this drug the largest initial placebo-controlled dataset of any of the oral PAH therapies; its once-daily dosing and excellent safety profile make it the most convenient of the therapies by a significant margin. After introducing the PAH disease state with references for more interested readers, this paper discusses the nitric oxide pathway as it relates to the pulmonary circulation, provides an overview of clinically available phosphodiesterase inhibitors and discusses tadalafil in relationship to sildenafil (Revatio®), the first phosphodiesterase inhibitor approved for PAH.

Sex-specific serum biomarker patterns in adults with Asperger's syndrome.

Autism spectrum conditions have been hypothesized to be an exaggeration of normal male low-empathizing and high-systemizing behaviors. We tested this hypothesis at the molecular level by performing comprehensive multi-analyte profiling of blood serum from adult subjects with Asperger's syndrome (AS) compared with controls. This led to identification of distinct sex-specific biomarker fingerprints for male and female subjects. Males with AS showed altered levels of 24 biomarkers including increased levels of cytokines and other inflammatory molecules. Multivariate statistical classification of males using this panel of 24 biomarkers revealed a marked separation between AS and controls with a sensitivity of 0.86 and specificity of 0.88. Testing this same panel in females did not result in a separation between the AS and control groups. In contrast, AS females showed altered levels of 17 biomarkers including growth factors and hormones such as androgens, growth hormone and insulin-related molecules. Classification of females using this biomarker panel resulted in a separation between AS and controls with sensitivities and specificities of 0.96 and 0.83, respectively, and testing this same panel in the male group did not result in a separation between the AS and control groups. The finding of elevated testosterone in AS females confirmed predictions from the 'extreme male brain' and androgen theories of autism spectrum conditions. We conclude that to understand the etiology and development of autism spectrum conditions, stratification by sex is essential.

Expression profiling of fibroblasts identifies cell cycle abnormalities in schizophrenia.

Many previous studies have attempted to gain insight into the underlying pathophysiology of schizophrenia by studying postmortem brain tissues of schizophrenia patients. However, such analyses can be confounded by artifactual features of this approach such as lengthy agonal state and postmortem interval times. As several aspects of schizophrenia are also manifested at the peripheral level in proliferating cell types, we have studied the disorder through systematic transcriptomic and proteomic analyses of skin fibroblasts biopsied from living patients. We performed comparative transcriptomic and proteomic profiling to characterize skin fibroblasts from schizophrenia patients compared to healthy controls. Transcriptomic profiling using cDNA array technology showed that pathways associated with cell cycle regulation and RNA processing were altered in the schizophrenia subjects (n = 12) relative to controls (n = 12). LC-MS(E) proteomic profiling led to identification of 16 proteins that showed significant differences in expression between schizophrenia (n = 11) and control (n = 11) subjects. Analysis in silico revealed that these proteins were also associated with proliferation and cell growth pathways. To validate these findings at the protein level, fibroblast protein extracts were analyzed by Western blotting which confirmed the differential expression of three key proteins associated with these pathways. At the functional level, we confirmed the decreased proliferation phenotype by showing that cultured fibroblasts from schizophrenia subjects (n = 5) incorporated less (3)H-thymidine into their nuclei compared to those from controls (n = 6) by day 4 over an 8 day time course study. Similar abnormalities in cell cycle and growth pathways have been reported to occur in the central nervous system in schizophrenia. These studies demonstrate that fibroblasts obtained from living schizophrenia subjects show alterations in cellular proliferation and growth pathways. Future studies aimed at characterizing such pathways in fibroblasts and other proliferating cell types from schizophrenia patients could elucidate the molecular mechanisms associated with the pathophysiology of schizophrenia and provide a useful model to support drug discovery efforts.

Global proteomic profiling reveals altered proteomic signature in schizophrenia serum.

Schizophrenia is one of the most severe psychiatric disorders affecting 1% of the world population. There is yet no empirical method to validate the diagnosis of the disease. The identification of an underlying molecular alteration could lead to an improved disease understanding and may yield an objective panel of biomarkers to aid in the diagnosis of this devastating disease. Presented is the largest reported liquid chromatography-mass spectrometry-based proteomic profiling study investigating serum samples taken from first-onset drug-naive patients compared with samples collected from healthy volunteers. The results of this large-scale study are presented along with enzyme-linked immunosorbent assay-based validation data.

Optimization of fast spiral chemical shift imaging using least squares reconstruction: application for hyperpolarized (13)C metabolic imaging.

A least-squares-based optimization and reconstruction algorithm has been developed for rapid metabolic imaging in the context of hyperpolarized (13)C. The algorithm uses a priori knowledge of resonance frequencies, J-coupling constants, and T(2)* values to enable acquisition of high-quality metabolic images with imaging times of approximately 100 ms for an 8-cm field of view (FOV) and 0.5 cm isotropic resolution. A root-mean-square error (rMSE) analysis is introduced to optimize metabolic image quality by appropriate choice of pulse sequence parameters, echo times, and signal model. By performing the reconstruction in k-space, the algorithm also allows the inclusion of the effect of chemical shift evolution during the readout period. Single-interleaf multiecho spiral chemical shift imaging (spCSI) is analyzed in detail as an illustrative example for the use of the new reconstruction and optimization algorithm. Simulation of the in vivo spectrum following the bolus injection of hyperpolarized (13)C(1) pyruvate shows that single-interleaf spiral spectroscopic imaging can achieve image quality in 100 ms, comparable to the performance of a 13-s phase-encoded chemical shift imaging (FIDCSI) experiment. Single-interleaf spCSI was also tested at a 3-T MR scanner using a phantom containing approximately 0.5-M solutions of alanine, lactate, and a pyruvate-pyruvate hydrate C(1)-C(2) ester at thermal equilibrium polarization, all enriched to 99% (13)C in the C(1) carbonyl positions. Upon reconstruction using the k-space-based least-squares technique, metabolite ratios obtained using the spCSI method were comparable to those obtained using a reference FIDCSI acquisition.

Two-stage evaluation and intervention program for control of methicillin-resistant Staphylococcus aureus in the hospital setting.

The aim of this study was to develop a 2-stage evaluation and intervention program for control of methicillin-resistant Staphylococcus aureus (MRSA) in the hospital setting. The first stage included evaluation of MRSA prevalence throughout the entire hospital; the presence of MRSA was determined in patients or medical staff who had a high risk of carrying it (i.e. as a result of contact with surgical wounds). In the second stage, "contact isolation" (which included the use of gloves, hand washing before and after treatment of a patient and isolation of patients' personal belongings) was carried out in every patient from whom MRSA was isolated in 4 intervention departments-Surgery, Orthopaedics, General ICU and Neonatal ICU-while the same policy of attempting to isolate MRSA was maintained. Both stages lasted 7 months. A comparison between MRSA prevalence in the evaluation and intervention stages disclosed a decrease in MRSA isolates from 91 to 56 in the entire hospital (p = 0.2) and from 45 to 24 in the intervention departments (p = 0.05), respectively; while the number of patients with MRSA decreased from 87 to 55 in the entire hospital (p = 0.2) and from 45 to 18 in the intervention departments (p = 0.007). The number of patients treated with vancomycin decreased from 48 before intervention to 23 after "contact isolation" was started in the entire hospital (p = 0.02) and from 31 to 5 in the intervention departments (p = 0.001). These results provide additional evidence in favor of establishing a program to control MRSA spread.