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Background: The mast cell-specific metalloprotease CPA3 has been given important roles in lung tissue homeostasis and disease pathogenesis. However, the dynamics and spatial distribution of mast cell CPA3 expression in lung diseases remain unknown., Methods: Using a histology-based approach for quantitative spatial decoding of mRNA and protein single cell, this study investigates the dynamics of CPA3 expression across mast cells residing in lungs from control subjects and patients with severe chronic obstructive pulmonary disease (COPD) or idiopathic lung fibrosis (IPF)., Results: Mast cells in COPD lungs had an anatomically widespread increase of CPA3 mRNA (bronchioles p < 0.001, pulmonary vessels p < 0.01, and alveolar parenchyma p < 0.01) compared to controls, while granule-stored CPA3 protein was unaltered. IPF lungs had a significant upregulation of both mast cell density, CPA3 mRNA ( p < 0.001) and protein ( p < 0.05), in the fibrotic alveolar tissue. Spatial expression maps revealed altered mast cell mRNA/protein quotients in lung areas subjected to disease-relevant histopathological alterations. Elevated CPA3 mRNA also correlated to lung tissue eosinophils, CD3 T cells, and declined lung function. Single-cell RNA sequencing of bronchial mast cells confirmed CPA3 as a top expressed gene with potential links to both inflammatory and protective markers., Conclusion: This study shows that lung tissue mast cell populations in COPD and IPF lungs have spatially complex and markedly upregulated CPA3 expression profiles that correlate with immunopathological alterations and lung function. Given the proposed roles of CPA3 in tissue homeostasis, remodeling, and inflammation, these alterations are likely to have clinical consequences., Competing Interests: Jimmie Jönsson is an employee of Medetect AB. Jonas S Erjefält is founder of Medetect AB. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be constructed as a potential conflict of interest., (Copyright © 2022 Siddhuraj, Jönsson, Alyamani, Prabhala, Magnusson, Lindstedt and Erjefält.)

Noninvasive optical imaging through dynamic scattering media has numerous important biomedical applications but still remains a challenging task. While standard diffuse imaging methods measure optical absorption or fluorescent emission, it is also well-established that the temporal correlation of scattered coherent light diffuses through tissue much like optical intensity. Few works to date, however, have aimed to experimentally measure and process such temporal correlation data to demonstrate deep-tissue video reconstruction of decorrelation dynamics. In this work, a single-photon avalanche diode array camera is utilized to simultaneously monitor the temporal dynamics of speckle fluctuations at the single-photon level from 12 different phantom tissue surface locations delivered via a customized fiber bundle array. Then a deep neural network is applied to convert the acquired single-photon measurements into video of scattering dynamics beneath rapidly decorrelating tissue phantoms. The ability to reconstruct images of transient (0.1-0.4 s) dynamic events occurring up to 8 mm beneath a decorrelating tissue phantom with millimeter-scale resolution is demonstrated, and it is highlighted how the model can flexibly extend to monitor flow speed within buried phantom vessels., (© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Fast noninvasive probing of spatially varying decorrelating events, such as cerebral blood flow beneath the human skull, is an essential task in various scientific and clinical settings. One of the primary optical techniques used is diffuse correlation spectroscopy (DCS), whose classical implementation uses a single or few single-photon detectors, resulting in poor spatial localization accuracy and relatively low temporal resolution. Here, we propose a technique termed C lassifying R apid decorrelation E vents via P arallelized single photon d E tection (CREPE) , a new form of DCS that can probe and classify different decorrelating movements hidden underneath turbid volume with high sensitivity using parallelized speckle detection from a 32 × 32 pixel SPAD array. We evaluate our setup by classifying different spatiotemporal-decorrelating patterns hidden beneath a 5 mm tissue-like phantom made with rapidly decorrelating dynamic scattering media. Twelve multi-mode fibers are used to collect scattered light from different positions on the surface of the tissue phantom. To validate our setup, we generate perturbed decorrelation patterns by both a digital micromirror device (DMD) modulated at multi-kilo-hertz rates, as well as a vessel phantom containing flowing fluid. Along with a deep contrastive learning algorithm that outperforms classic unsupervised learning methods, we demonstrate our approach can accurately detect and classify different transient decorrelation events (happening in 0.1-0.4 s) underneath turbid scattering media, without any data labeling. This has the potential to be applied to non-invasively monitor deep tissue motion patterns, for example identifying normal or abnormal cerebral blood flow events, at multi-Hertz rates within a compact and static detection probe., Competing Interests: SX and RH have submitted a patent application related to this work, assigned to Duke University. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xu, Liu, Yang, Jönsson, Qian, McKee, Kim, Konda, Zhou, Kreiß, Wang, Berrocal, Huettel and Horstmeyer.)

Human papillomavirus type 16 (HPV16) E2 is an essential HPV16 protein. We have investigated how HPV16 E2 expression is regulated and have identifed a splicing enhancer that is required for production of HPV16 E2 mRNAs. This uridine-less splicing enhancer sequence (ACGAGGACGAGGACAAGGA) contains 84% adenosine and guanosine and 16% cytosine and consists of three 'AC(A/G)AGG'-repeats. Mutational inactivation of the splicing enhancer reduced splicing to E2-mRNA specific splice site SA2709 and resulted in increased levels of unspliced E1-encoding mRNAs. The splicing enhancer sequence interacted with cellular RNA binding protein hnRNP G that promoted splicing to SA2709 and enhanced E2 mRNA production. The splicing-enhancing function of hnRNP G mapped to amino acids 236-286 of hnRNP G that were also shown to interact with splicing factor U2AF65. The interactions between hnRNP G and HPV16 E2 mRNAs and U2AF65 increased in response to keratinocyte differentiation as well as by the induction of the DNA damage response (DDR). The DDR reduced sumoylation of hnRNP G and pharmacological inhibition of sumoylation enhanced HPV16 E2 mRNA splicing and interactions between hnRNP G and E2 mRNAs and U2AF65. Intriguingly, hnRNP G also promoted intron retention of the HPV16 E6 coding region thereby inhibiting production of spliced E7 oncogene mRNAs., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

There is a growing awareness that the large number of environmental pollutants we are exposed to on a daily basis are causing major health problems. Compared to traditional studies that focus on individual pollutants, there are relatively few studies on how pollutants mixtures interact. Several studies have reported a relationship between environmental pollutants and the development of cancer, even when pollutant levels are below toxicity reference values. The possibility of synergistic interactions between different pollutants could explain how even low concentrations can cause major health problems. These intricate that molecular interactions can occur through a wide variety of mechanisms, and our understanding of the physiological effects of mixtures is still limited. The purpose of this paper is to discuss recent reports that address possible synergistic interactions between different types of environmental pollutants that could promote cancer development. Our literature studies suggest that key biological pathways are frequently implicated in such processes. These include increased production of reactive oxygen species, activation by cytochrome P450, and aryl hydrocarbon receptor signaling, among others. We discuss the need to understand individual pathological vulnerability not only in relation to basic genetics and gene expression, but also in terms of measurable exposure to contaminants. We also mention the need for significant improvements in future studies using a multitude of disciplines, such as the development of high-throughput study models, better tools for quantifying pollutants in cancer patients, innovative pharmacological and toxicological studies, and high-efficiency computer analysis, which allow us to analyze the molecular mechanisms of mixtures., Competing Interests: The authors declare no conflicts of interest relevant to this study., (© 2022 The Authors. GeoHealth published by Wiley Periodicals LLC on behalf of American Geophysical Union.)

We have determined the effect of seven serine- and arginine-rich (SR) proteins and 15 heterogenous nuclear ribonucleoproteins (hnRNPs) on human papillomavirus type 16 (HPV16) late gene expression. Of the seven SR proteins analyzed here, SRSF1, SRSF3, and SRSF9 induced HPV16 late gene expression, and five of the SR proteins affected HPV16 L1 mRNA splicing. Of the 15 hnRNP proteins analyzed here, hnRNP A2, hnRNP F, and hnRNP H efficiently induced HPV16 late gene expression, and all of the hnRNPs affected HPV16 L1 mRNA levels or mRNA splicing. Thus, the majority of SR proteins and hnRNPs have the potential to regulate HPV16 L1 mRNA splicing. Strict control of the expression of the immunogenic L1 and L2 capsid proteins may contribute to the ability of HPV16 to cause persistence., (© 2021. The Author(s).)

This article, Part II of an article series on GPU-accelerated Monte Carlo simulation of photon transport through turbid media, focuses on the validation of the online software Multi-Scattering. While Part I detailed the implementation of the computational model, simulated and experimental results are now compared for the distribution of the scattered light intensity. The scattering phantoms prepared here are aqueous dispersions of polystyrene microspheres of diameter D = 0.5, 2 and 5 μm and at various concentrations, resulting in optical depth ranging from OD = 1 to 17.5. The Lorenz-Mie scattering phase functions used in the simulations have been verified experimentally at low particle concentrations by analyzing the angular light intensity distribution at the Fourier plane of a collecting lens. The validation approach herein accounts for the specific light collection and image formation by the camera. The front and side surfaces of the medium are imaged and the corresponding light intensity distributions are compared qualitatively and quantitatively. It is concluded that the model enables reliable simulations over the tested parameters, offering predictive simulations of transmitted intensities with a mean relative error ≤~19% over the full range. The online software is available at: https://multi-scattering.com/.

C-reactive protein (CRP) is an inflammation biomarker that should be quantified accurately during infections and healing processes. Nanobodies are good candidates to replace conventional antibodies in immunodiagnostics due to their inexpensive production, simple engineering, and the possibility to obtain higher binder density on capture surfaces. Starting from the same pre-immune library, we compared the selection output resulting from two independent panning strategies, one exclusively exploiting the phage display and another in which a first round of phage display was followed by a second round of yeast display. There was a partial output convergence between the two methods, since two clones were identified using both panning protocols but the first provided several further different sequences, whereas the second favored the recovery of many copies of few clones. The isolated anti-CRP nanobodies had affinity in the low nanomolar range and were suitable for ELISA and immunoprecipitation. One of them was fused to SpyTag and exploited in combination with SpyCatcher as the immunocapture element to quantify CRP using electrochemical impedance spectroscopy. The sensitivity of the biosensor was calculated as low as 0.21 μg/mL.

This paper reports the synthesis of high-quality upconverting nanoparticles (UCNPs) - sodium yttrium tetrafluoride doped with ytterbium and erbium (NaYF 4 :Yb,Er) with a silica shell and capped with phenyl functional groups. The main goal of this research was to design tailor-made UCNPs for fingermark detection, to test and validate a nanoparticle-based detection technique and to compare their performance against a benchmark method to assess potential implementation in routine practice by law enforcement agencies. The water-based UCNPs solution was applied to natural fingermarks on a number of substrates. This is the first ever systematic comparative study between UCNPs and a benchmark fingermark detection technique - cyanoacrylate fuming (CAF) followed by luminescent dye staining. Fingermark detection effectiveness was studied by treating 300 latent fingermark specimens on aluminium foil, polyethylene, polypropylene and glass slides. It was concluded that, on average, CAF performed better across the substrates tested. Nevertheless, UCNPs can be advantageous for fingermark detection on multicoloured, patterned or luminescent substrates due to their unique optical properties. There are, however, shortfalls associated with their synthesis and use that need to be addressed before they can be considered for operational purposes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objective: To combine prospective cohort studies, by including HLA harmonization, and estimate risk of islet autoimmunity and progression to clinical diabetes., Research Design and Methods: For prospective cohorts in Finland, Germany, Sweden, and the U.S., 24,662 children at increased genetic risk for development of islet autoantibodies and type 1 diabetes have been followed. Following harmonization, the outcomes were analyzed in 16,709 infants-toddlers enrolled by age 2.5 years., Results: In the infant-toddler cohort, 1,413 (8.5%) developed at least one autoantibody confirmed at two or more consecutive visits (seroconversion), 865 (5%) developed multiple autoantibodies, and 655 (4%) progressed to diabetes. The 15-year cumulative incidence of diabetes varied in children with one, two, or three autoantibodies at seroconversion: 45% (95% CI 40-52), 85% (78-90), and 92% (85-97), respectively. Among those with a single autoantibody, status 2 years after seroconversion predicted diabetes risk: 12% (10-25) if reverting to autoantibody negative, 30% (20-40) if retaining a single autoantibody, and 82% (80-95) if developing multiple autoantibodies. HLA-DR-DQ affected the risk of confirmed seroconversion and progression to diabetes in children with stable single-autoantibody status. Their 15-year diabetes incidence for higher- versus lower-risk genotypes was 40% (28-50) vs. 12% (5-38). The rate of progression to diabetes was inversely related to age at development of multiple autoantibodies, ranging from 20% per year to 6% per year in children developing multipositivity in ≤2 years or >7.4 years, respectively., Conclusions: The number of islet autoantibodies at seroconversion reliably predicts 15-year type 1 diabetes risk. In children retaining a single autoantibody, HLA-DR-DQ genotypes can further refine risk of progression., (© 2021 by the American Diabetes Association.)

Alkyl glycoside surfactants with elongated carbohydrate chains are useful in different applications due to their improved biocompatibility. Cyclodextrin glucanotransferases can catalyze the elongation process through the coupling reaction. However, due to the presence of a hydrophobic tail, the interaction between an alkyl glycoside acceptor and the active site residues is weaker than the interaction with maltooligosaccharides at the corresponding site. Here we report the mutations of F197, G263 and E266 near the acceptor subsites in the CGTase CspCGT13 from Carboxydocella sp. The results showed that substitutions of both F197 and G263 were important for the binding of acceptor substrate dodecyl maltoside during coupling reaction. The double mutant F197Y/G263A showed enhanced coupling activity and displayed a 2-fold increase of the primary coupling product using γ-cyclodextrin as donor when compared to wildtype CspCGT13. Disproportionation activity was also reduced, which was also the case for another double mutant (F197Y/E266A) that however not showed the corresponding increase in coupling. A triple mutant F197Y/G263A/E266A maintained the increase in primary coupling product (1.8-fold increase) using dodecyl maltoside as acceptor, but disproportionation was approximately at the same level as in the double mutants. In addition, hydrolysis of starch was slightly increased by the F197Y and G263A substitutions, indicating that interactions at both positions influenced the selectivity between glycosyl and alkyl moieties., (© The Author(s) 2020. Published by Oxford University Press.)

Objectives: To investigate whether the association between pain intensity and willingness to undergo surgery is explained by walking difficulties, in patients with knee or hip osteoarthritis (OA)., Methods: This is an observational study using data from the Better management of patients with Osteoarthritis (BOA) register, which collects data from a publicly financed self-management programme for people with OA in Sweden. We included all patients with knee or hip OA who attended the baseline visit between 2008 and 2016. We conducted separate mediation analyses within a counterfactual framework to estimate the mediation effect of walking difficulties (yes/no) on willingness to undergo surgery (yes/no) for each one-point increase in pain (0-10 on a numeric rating scale), adjusted for relevant confounders., Results: We included 72,131 patients (69% women, mean age 66, mean pain 5.4, 81% had walking difficulties, 27% was willing to undergo surgery). A one-point increase in pain intensity was associated with 1.53 (95% CI: 1.51; 1.55) higher odds of being willing to undergo surgery. Walking difficulties mediated 10%-25% of the effect of one-point increase in pain when pain was <8/10, while at pain ≥8/10 this percentage decreased to 3%., Conclusions: More than 80% of the BOA patients have mild to moderate pain (<8/10) and walking difficulties can mediate up to a quarter of the total effect of pain on the willingness to undergo surgery in these patients. Trials to evaluate the potential to lower surgery demand by reducing walking difficulties in people with these characteristics are needed., Competing Interests: This manuscript describes original work and is not under consideration by any other journal. All authors approved the manuscript and this submission. Thérése Jönsson is part of the steering committee of the BOA registry, for which she does not receive any compensation. Leif Dahlberg is the Co-founder and Chief Medical Officer of Joint Academy, a company which provides web-based non-surgical interventions for patients with hip and knee osteoarthritis. All other authors report no competing interests., (© 2021 The Author(s).)

Maternal obesity may lead to epigenetic alterations in the offspring and might thereby contribute to disease later in life. We investigated whether a lifestyle intervention in pregnant women with obesity is associated with epigenetic variation in cord blood and body composition in the offspring. Genome-wide DNA methylation was analyzed in cord blood from 208 offspring from the Treatment of Obese Pregnant women (TOP)-study, which includes pregnant women with obesity randomized to lifestyle interventions comprised of physical activity with or without dietary advice versus control subjects (standard of care). DNA methylation was altered at 379 sites, annotated to 370 genes, in cord blood from offspring of mothers following a lifestyle intervention versus control subjects (false discovery rate [FDR] <5%) when using the Houseman reference-free method to correct for cell composition, and three of these sites were significant based on Bonferroni correction. These 370 genes are overrepresented in gene ontology terms, including response to fatty acids and adipose tissue development. Offspring of mothers included in a lifestyle intervention were born with more lean mass compared with control subjects. Methylation at 17 sites, annotated to, for example, DISC1 , GBX2 , HERC2 , and HUWE1 , partially mediates the effect of the lifestyle intervention on lean mass in the offspring (FDR <5%). Moreover, 22 methylation sites were associated with offspring BMI z scores during the first 3 years of life ( P < 0.05). Overall, lifestyle interventions in pregnant women with obesity are associated with epigenetic changes in offspring, potentially influencing the offspring's lean mass and early growth., (© 2021 by the American Diabetes Association.)

Hydrophobicity is one of the most critical factors governing the adsorption of molecules and objects, such as virions, on surfaces. Even moderate change of wetting angle of plastic surfaces causes a drastic decrease ranging from 2 to 5 logs of the viruses (e.g., T4 phage) in the suspension due to adsorption on polymer vials' walls. The effect varies immensely in seemingly identical containers but purchased from different vendors. Comparison of glass, polyethylene, polypropylene, and polystyrene containers revealed a threshold in the wetting angle of around 95°: virions adsorb on the surface of more hydrophobic containers, while in more hydrophilic vials, phage suspensions are stable. The polypropylene surface of the Eppendorf-type and Falcon-type can accommodate from around 10 8 PFU/ml to around 10 10 PFU/ml from the suspension. The adsorption onto the container's wall might result in complete scavenging of virions from the bulk. We developed two methods to overcome this issue. The addition of surfactant Tween20 and/or plasma treatment provides a remedy by modulating surface wettability and inhibiting virions' adsorption. Plastic containers are essential consumables in the daily use of many bio-laboratories. Thus, this is important not only for phage-related research (e.g., the use of phage therapies as an alternative for antibiotics) but also for data comparison and reproducibility in the field of biochemistry and virology.

Silver nanowires with varying diameters and submillimeter lengths were obtained by changing a reducing agent used during hydrothermal synthesis. The control over the nanowire diameter turns out to play a critical role in determining their plasmonic properties, including fluorescence enhancement and surface plasmon polariton propagation. Advanced fluorescence imaging of hybrid nanostructures assembled of silver nanowires and photoactive proteins indicates longer propagation lengths for nanowires featuring larger diameters. At the same time, with increasing diameter of the nanowires, we measure a substantial reduction of fluorescence enhancement. The results point at possible ways to control the influence of plasmon excitations in silver nanowires by tuning their morphology.

In this article we present and describe an online freely accessible software called Multi-Scattering for the modeling of light propagation in scattering and absorbing media. Part II of this article series focuses on the validation of the model by rigorously comparing the simulated results with experimental data. The model is based on the use of the Monte Carlo method, where billions of photon packets are being tracked through simulated cubic volumes. Simulations are accelerated by the use of general-purpose computing on graphics processing units, reducing the computation time by a factor up to 200x in comparison with a single central processing unit thread. By using four graphic cards on a single computer, the simulation speed increases by a factor of 800x. For an anisotropy factor g = 0.86, this enables the transport path of one billion photons to be computed in 10 seconds for optical depth OD = 10 and in 20 minutes for OD = 500. Another feature of Multi-Scattering is the integration and implementation of the Lorenz-Mie theory in the software to generate the scattering phase functions from spherical particles. The simulations are run from a computer server at Lund University, allowing researchers to log in and use it freely without any prior need for programming skills or specific software/hardware installations. There are countless types of scattering media in which this model can be used to predict light transport, including medical tissues, blood samples, clouds, smoke, fog, turbid liquids, spray systems, etc. An example of simulation results is given here for photon propagation through a piece of human head. The software also includes features for modeling image formation by inserting a virtual collecting lens and a detection matrix which simulate a camera objective and a sensor array respectively. The user interface for setting-up simulations and for displaying the corresponding results is found at: https://multi-scattering.com/.

We introduce a new latent fingermark (LFM) development method, where compounds showing long lifetime luminescence are generated in situ by the reactions of Eu(TTA) 3 (H 2 O) 2 with LFM components. Until now, time-gated imaging could not be used to develop LFM on porous surfaces due to the difficulties with selective binding of the developing agents to the fingermark ridges. The nature of the interactions of Eu(TTA) 3 (H 2 O) 2 with the LFM material has been investigated for three model compounds commonly found in the LFM composition-oleic acid, l-serine, and squalene. The LFMs developed with the europium β-diketonate complex have been successfully photographed using a time-gated imaging scheme. The presented new approach has been demonstrated to give similar or better results than developing agents commonly used for paper samples (ninhydrin and 1,2-indanedione). Moreover, contrary to the methods mentioned above, the new approach allows for the development of amino acid-poor LFM on paper.

Recently, bacteriophage particles have started to be applied as a new biomaterial for developing sensing platforms. They can be used as both a recognition element or/and as building blocks, template/scaffold. In this paper, we studied a bacteriophage selected through phage-display technology. The chosen bacteriophage acted as a building block for creating a carbon nanofiber-based electrode and as a new receptor/binding element that recognizes C-reactive protein (CRP) - one of the markers of inflammatory processes in the human body. The binding efficiency of the selected phage towards CRP is two orders of magnitude higher than in the wild type. We demonstrate that the phage-based sensor is selective against other proteins. Finally, we show that layer-by-layer methods are suitable for deposition of negatively charged phages (wild or CRP-binding) with positively charged carbon nanofibers for electrode surface modification. A three-layered electrode was successfully used for molecular recognition of CRP, and the molecular interactions were studied using electrochemical, biological, and optical methods, including microscopic and spectroscopic analyses., Competing Interests: Declaration of Competing Interest The authors declared that there is no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We demonstrate that single functionalized silver nanowires form a geometric platform suitable for efficient real-time detection of single photoactive proteins. By collecting series of images using wide-field fluorescence microscopy, events of single protein attachment can be distinguished with the signal to noise ratio further improved by fluorescence enhancement due to plasmon excitations in the nanowires. The enhancement is evidenced by strong shortening of the fluorescence decay of single photoactive proteins conjugated to the silver nanowires.

Human papillomavirus 16 (HPV16) 5'-splice site SD226 and 3'-splice site SA409 are required for production of the HPV16 E7 mRNAs, whereas unspliced mRNAs produce E6 mRNAs. The E6 and E7 proteins are essential in the HPV16 replication cycle but are also the major HPV16 proteins required for induction and maintenance of malignancy caused by HPV16 infection. Thus, a balanced expression of unspliced and spliced mRNAs is required for production of sufficient quantities of E6 and E7 proteins under physiological and pathophysiological conditions. If splicing becomes too efficient, the levels of unspliced E6 mRNAs will decrease below a threshold level that is no longer able to produce E6 protein quantities high enough to significantly reduce p53 protein levels. Similarly, if splicing becomes too inefficient, the levels of spliced E7 mRNAs will decrease below a threshold level that is no longer able to produce E7 protein quantities high enough to significantly reduce pRb protein levels. To determine how splicing between SD226 and SA409 is regulated, we have investigated how SA409 is controlled by the cellular proteins hnRNP A1 and hnRNP A2, two proteins that have been shown previously to control HPV16 gene expression. We found that hnRNP A1 and A2 interacted directly and specifically with a C-less RNA element located between HPV16 nucleotide positions 594 and 604 downstream of SA409. Overexpression of hnRNP A1 inhibited SA409 and promoted production of unspliced E6 mRNAs at the expense of the E7 mRNAs, whereas overexpression of hnRNP A2 inhibited SA409 to redirect splicing to SA742, a downstream 3'-splice site that is used for generation of HPV16 E6̂E7, E1, and E4 mRNAs. Thus, high levels of either hnRNP A1 or hnRNP A2 inhibited production of the promitotic HPV16 E7 protein. We show that the hnRNP A1 and A2 proteins control the relative levels of the HPV16 unspliced and spliced HPV16 E6 and E7 mRNAs and function as inhibitors of HPV16 E7 expression. IMPORTANCE Human papillomavirus type 16 (HPV16) belongs to the high-risk-group of HPVs and is causing a variety of anogenital cancers and head and neck cancer. The two HPV16 oncoproteins E6 and E7 prevent apoptosis and promote mitosis and are essential for completion of the HPV16 life cycle and for transformation of the infected cell and maintenance of malignancy. E6 and E7 are produced from two mRNAs that are generated in a mutually exclusive manner by alternative splicing. While E6 protein is made from the unspliced mRNA, E7 is made from the spliced version of the same pre-mRNA. Since sufficient quantities of both E6 and E7 are required for malignant transformation, this intricate arrangement of gene expression renders E6 and E7 expression vulnerable to external interference. Since antiviral drugs to HPV16 are not available, a detailed knowledge of the regulation of HPV16 E6 and E7 mRNA splicing may uncover novel targets for therapy., (Copyright © 2020 Zheng et al.)

Infection patterns of avian haemosporidians result from the evolution of their associations with hosts, and can be shaped by multiple biotic factors. However, at the level of parasite species, few studies have investigated the details of the temporal dynamics of infection patterns in wild bird communities. We hereby studied a wild bird community in southern Sweden to investigate two generalist parasites (cyt b lineages PARUS1 and WW2) of the morphological species Haemoproteus majoris in their main host species (tits and warblers, respectively) to look for seasonal (spring to autumn) and age class related variation in infection patterns. For both lineages, we detected a similar temporal pattern in prevalence and infection intensity, with peak levels during the main nesting season in adults and a few weeks later in juveniles. Infections in juveniles were detected as soon as they started to be caught by mist nets, implying that they became infected when still in the nest or during the first weeks post-fledging. The initially high intensities in juveniles were followed by a significant decrease during the hatching year, emphasising the importance of studying haemosporidian infections in nestlings and fledglings. Both prevalence and infection intensity in adults increased from spring to early summer, either due to spring relapses or new infections. Both prevalence and infection intensity declined in adults at the time when independent juveniles of the respective species started to appear, suggesting that the rate of parasite withdrawal from blood exceeded the rates of new infections gained and relapses of previous infections. Prevalence in both juveniles and adults approached zero towards the end of the summer., (Copyright © 2020 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Although elevated blood or sputum eosinophils are present in many patients with COPD, uncertainties remain regarding the anatomical distribution pattern of lung-infiltrating eosinophils. Basophils have remained virtually unexplored in COPD. This study mapped tissue-infiltrating eosinophils, basophils and eosinophil-promoting immune mechanisms in COPD-affected lungs.Surgical lung tissue and biopsies from major anatomical compartments were obtained from COPD patients with severity grades Global Initiative for Chronic Obstructive Lung Disease stages I-IV; never-smokers/smokers served as controls. Automated immunohistochemistry and in situ hybridisation identified immune cells, the type 2 immunity marker GATA3 and eotaxins (CCL11, CCL24).Eosinophils and basophils were present in all anatomical compartments of COPD-affected lungs and increased significantly in very severe COPD. The eosinophilia was strikingly patchy, and focal eosinophil-rich microenvironments were spatially linked with GATA3 + cells, including type 2 helper T-cell lymphocytes and type 2 innate lymphoid cells. A similarly localised and interleukin-33/ST2-dependent eosinophilia was demonstrated in influenza-infected mice. Both mice and patients displayed spatially confined eotaxin signatures with CCL11 + fibroblasts and CCL24 + macrophages.In addition to identifying tissue basophilia as a novel feature of advanced COPD, the identification of spatially confined eosinophil-rich type 2 microenvironments represents a novel type of heterogeneity in the immunopathology of COPD that is likely to have implications for personalised treatment., Competing Interests: Conflict of interest: P. Jogdand has nothing to disclose. Conflict of interest: P. Siddhuraj has nothing to disclose. Conflict of interest: M. Mori has nothing to disclose. Conflict of interest: C. Sanden has nothing to disclose. Conflict of interest: J. Jönsson has nothing to disclose. Conflict of interest: A.F. Walls has nothing to disclose. Conflict of interest: J. Kearley is an employee of AstraZeneca (formerly MedImmune LLC) and has stock options in AstraZeneca. Conflict of interest: A.A. Humbles was an employee of AstraZeneca (formerly MedImmune LLC) at the time these analyses were conducted. Conflict of interest: R. Kolbeck was an employee of AstraZeneca (formerly MedImmune LLC) at the time these analyses were conducted. Conflict of interest: L. Bjermer has nothing to disclose. Conflict of interest: P. Newbold is an employee of AstraZeneca (formerly MedImmune LLC) and has stock options in AstraZeneca. Conflict of interest: J.S. Erjefält is founder (and stock owner) of Medetect AB, who received funding from AstraZeneca for conducting parts of the present study., (Copyright ©ERS 2020.)

In this work we discuss a new label-free biosensing device based on indium tin oxide (ITO) overlaid section of a multimode optical fiber fused silica core. The sensor has been used to optical measurements also simultaneously interrogated electrochemically (EC). Due to optimized thickness and optical properties of ITO film, a lossy-mode resonance (LMR) could be observed in the optical domain, where electrical properties of the film allowed for application of the sensor as a working electrode in an EC setup. It has been confirmed that the LMR response depends on optical properties of the external medium, as well as potential applied to the electrode during cyclic voltammetry. After the ITO surface functionalization with amine groups and covalently attached biotin, the device has been applied for label-free biosensing of avidin in both the domains simultaneously. On the example of biotin-avidin detection system it was demonstrated that when avidin concentration increases a decrease in current and increase in LMR wavelength shift were recorded in EC and optical domain, respectively. Both optical and EC responses follow the protein interaction process, and thus can be used as cross-verification of the readouts. Moreover, an extended information has been achieved comparing to solely EC interrogation, i.e., the grafting process of biotin and avidin was directly monitored optically displaying individual steps of an incubation procedure., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this paper, we demonstrate plasmonic substrates prepared on demand, using a straightforward technique, based on laser-induced photochemical reduction of silver compounds on a glass substrate. Importantly, the presented technique does not impose any restrictions regarding the shape and length of the metallic pattern. Plasmonic interactions have been probed using both Stokes and anti-Stokes types of emitters that served as photoluminescence probes. For both cases, we observed a pronounced increase of the photoluminescence intensity for emitters deposited on silver patterns. By studying the absorption and emission dynamics, we identified the mechanisms responsible for emission enhancement and the position of the plasmonic resonance.

In this work we demonstrate a composite material based on silica particles. The particles have been doped with zinc oxide quantum dots which possess long living luminescence. The surface of the particles has been functionalized with phenyl groups using sol-gel process. The new material has been successfully applied for visualization of natural latent fingermarks on several surfaces, in particular, those showing their own luminescence and intensive background staining while using powder dusting, what is of the vital interest of forensic science. The time-gated imaging allows to overcome the background luminescence problem and surface functionalization increases the affinity of the particles to the fingermarks, what improves the selectivity of a new developing agent. The main novelty of the presented approach is the use of composite material that combines two main features-long lifetime luminescence and the ability to preferentially attach to the fingermark, due to hydrophobic interactions. Moreover,the utilization of deposition from the suspension instead of simple powder dusting allows for development of latent fingermarks on the surfaces that are difficult to work with powders (e.g. sticky side of the adhesive tape).

Since the norovirus is the main cause of acute gastroenteritis all over the world, its fast detection is crucial in medical diagnostics. In this work, a rapid, sensitive, and selective optical fiber biosensor for the detection of norovirus virus-like particles (VLPs) is reported. The sensor is based on highly sensitive long-period fiber gratings (LPFGs) coated with antibodies against the main coat protein of the norovirus. Several modification methods were verified to obtain reliable immobilization of protein receptors on the LPFG surface. We were able to detect 1 ng/mL norovirus VLPs in a 40-min assay in a label-free manner. Thanks to the application of an optical fiber as the sensor, there is a possibility to increase the user's safety by separating the measurement point from the signal processing setup. Moreover, our sensor is small and light, and the proposed assay is straightforward. The designed LPFG-based biosensor could be applied in both fast norovirus detection and in vaccine testing.

Type 2 diabetes (T2D) is characterized by insufficient insulin secretion and elevated glucose levels, often in combination with high levels of circulating fatty acids. Long-term exposure to high levels of glucose or fatty acids impair insulin secretion in pancreatic islets, which could partly be due to epigenetic alterations. We studied the effects of high concentrations of glucose and palmitate combined for 48 h (glucolipotoxicity) on the transcriptome, the epigenome, and cell function in human islets. Glucolipotoxicity impaired insulin secretion, increased apoptosis, and significantly (false discovery rate <5%) altered the expression of 1,855 genes, including 35 genes previously implicated in T2D by genome-wide association studies (e.g., TCF7L2 and CDKN2B ). Additionally, metabolic pathways were enriched for downregulated genes. Of the differentially expressed genes, 1,469 also exhibited altered DNA methylation (e.g., CDK1 , FICD , TPX2 , and TYMS ). A luciferase assay showed that increased methylation of CDK1 directly reduces its transcription in pancreatic β-cells, supporting the idea that DNA methylation underlies altered expression after glucolipotoxicity. Follow-up experiments in clonal β-cells showed that knockdown of FICD and TPX2 alters insulin secretion. Together, our novel data demonstrate that glucolipotoxicity changes the epigenome in human islets, thereby altering gene expression and possibly exacerbating the secretory defect in T2D., (© 2019 by the American Diabetes Association.)

Interactions with the microbiota influence many aspects of immunity, including immune cell development, differentiation, and function. Here, we examined the impact of the microbiota on CD8 + T cell memory. Antigen-activated CD8 + T cells transferred into germ-free mice failed to transition into long-lived memory cells and had transcriptional impairments in core genes associated with oxidative metabolism. The microbiota-derived short-chain fatty acid (SCFA) butyrate promoted cellular metabolism, enhanced memory potential of activated CD8 + T cells, and SCFAs were required for optimal recall responses upon antigen re-encounter. Mechanistic experiments revealed that butyrate uncoupled the tricarboxylic acid cycle from glycolytic input in CD8 + T cells, which allowed preferential fueling of oxidative phosphorylation through sustained glutamine utilization and fatty acid catabolism. Our findings reveal a role for the microbiota in promoting CD8 + T cell long-term survival as memory cells and suggest that microbial metabolites guide the metabolic rewiring of activated CD8 + T cells to enable this transition., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The aim was to describe school nurses' experiences working with students with mental health problems. In this inductive qualitative study, interviews were conducted with 14 school nurses in Sweden. The content analysis revealed three themes: (1) sense of worriedness about working with students with mental health problems, (2) taking care of students with mental health issues was an opportunity for personal and professional development, and (3) the experience of making a difference for young people with mental health problems. The school nurses working with students who have mental health problems had to cope with their own emotions, worries, and feelings of insufficiency. However, the school nurses also found the work to be meaningful and rewarding because they appreciated the opportunity for personal and professional development while taking care of students with mental health problems. They felt grateful for having a profession that had a huge impact on children's/adolescents' lives.

The purpose of this review is to introduce and present the concept of metallic nanowires as building-blocks of plasmonically active structures. In addition to concise description of both the basic physical properties associated with the electron oscillations as well as energy propagation in metallic nanostructures, and methods of fabrication of metallic nanowires, we will demonstrate several key ideas that involve interactions between plasmon excitations and electronic states in surrounding molecules or other emitters. Particular emphasis will be placed on the effects that involve not only plasmonic enhancement or quenching of fluorescence, but also propagation of energy on lengths that exceed the wavelength of light.

We report on the synthesis of long silver nanowires using the hydrothermal method, with H₂O₂ as the reducing agent. Our approach yields nanowires with an average diameter and length of about 100 nm and 160 µm, respectively, reaching the maximum length of 800 µm. Scanning electron microscopy (SEM) measurements revealed the presence of a thick, inhomogeneous poly(vinylpyrrolidone) (PVP) layer covering the nanowires, which with time becomes much more uniform, leading to well-defined extinction peaks in the ultraviolet-visible (UV-Vis) spectra. This change in morphology is evidenced also by the fluorescence enhancement behavior probed using protein complexes. Wide-field and confocal fluorescence microscopy measurements demonstrate strong, 10-fold enhancement of the protein emission intensity, accompanied by a reduction of the fluorescence decay time. In addition, for the aged, one-month-old nanowires, the uniformity of the intensity profile along them was substantially improved as compared with the as-synthesized ones. The results point towards the importance of the morphology of plasmonically active silver nanowires when considering their application in enhancing optical properties or achieving energy propagation over submillimeter distances.

Parasites that can infect multiple host species are considered to be host generalists with low host specificity. However, whether generalist parasites are better adapted to a subset of their host species remains unknown. To elucidate this possibility, we compared the variation in prevalence and infection intensity among host species of three generalist parasite lineages belonging to the morphological species Haemoproteus majoris, in a natural bird community in southern Sweden. Prevalence in each host species was confirmed by nested PCR and DNA sequencing, and infection intensities were quantified using lineage-specific real-time qPCR. For two of the three lineages, we detected positive correlations between prevalence and infection intensity, indicating that these generalist parasites are better adapted to a subset of host species, which may have been more frequently encountered during the evolution of the parasite; we refer to these as main host species. For both lineages, the main host species were more phylogenetically related than expected by chance as revealed by strong phylogenetic signal in prevalence among hosts. By comparing our results with previous records of these parasites, we found that the host range of a generalist parasite can vary among different communities and may partly be shaped by the presence of other parasites. Our study reveals that generalist parasites may be specialized on a subset of their host species and it highlights the importance of considering infection intensity and host phylogeny when determining the host specificity of a parasite., (© 2018 John Wiley & Sons Ltd.)

The interaction between the T4 bacteriophage gp37 adhesin and the bacterial lipopolysaccharide (LPS) is a well-studied system, however, the affinity and strength of the interaction haven't been analyzed so far. Here, we use atomic force microscopy to determine the strength of the interaction between the adhesin and its receptor, namely LPS taken from a wild strain of E. coli B. As negative controls we used LPSs of E. coli O111:B and Hafnia alvei. To study the interaction an AFM tip modified with the gp37 adhesin was used to scan surfaces of mica covered with one of the three different LPSs. Using the correlation between the surface topography images and the tip-surface interaction we could verify the binding between the specific LPS and the tip in contrast to the very weak interaction between the tip and the non-binding LPSs. Using force spectroscopy we could then measure the binding strength by pulling on the AFM tip until it lifted off from the surface. The force necessary to break the interaction between gp37 and LPS from E. coli B, LPS from E. coli O111:B and LPS from H. alvei were measured to be 70 ± 29 pN, 46 ± 13 pN and 45 ± 14 pN, respectively. The latter values are likely partially due to non-specific interaction between the gp37 and the solid surface, as LPS from E. coli O111:B and LPS from H. alvei have been shown to not bind to gp37, which is confirmed by the low correlation between binding and topography for these samples.

Solstice ® Performance Fluid (PF), trans-1-chloro-3,3,3-trifluoropropene, is presented as an alternative to HFE7100, methoxy-nonafluorobutane, as a carrier solvent in a number of chemical formulations used for the visualisation of latent fingermarks. The supply of HFE7100 may be at risk due to a recent European Union regulation to control global warming. Laboratory trials using split depletions and a pseudo-operational trial of 1000 porous samples have shown that Solstice ® PF is a viable alternative to HFE7100 for the chemical formulations of ninhydrin and 1,2-indanedione. Other preliminary trials have also indicated that Solstice ® PF can be used as a carrier solvent for the zinc toning of marks found using ninhydrin as well as the α-naphtholflavone fixative solution for iodine developed marks. Results from the pseudo-operational trial demonstrate that the number of marks detected by ninhydrin and 1,2-indanedione formulations for each carrier solvent is comparable. When compared to HFE7100, advantages of Solstice ® PF include a very low global warming potential and atmospheric lifetime in addition to a higher wetting index and lower costs. This study also provides a validation study that supports the potential replacement of DFO with 1,2-indanedione., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Fast and reliable bacteria detection is crucial for lowering the socioeconomic burden related to bacterial infections (e.g., in healthcare, industry or security). Bacteriophages (i.e., viruses with bacterial hosts) pose advantages such as great specificity, robustness, toughness and cheap preparation, making them popular biorecognition elements in biosensors and other assays for bacteria detection. There are several possible designs of bacteriophage-based biosensors. Here, we focus on developments based on whole virions as recognition agents. We divide the review into sections dealing with phage lysis as an analytical signal, phages as capturing elements in assays and phage-based sensing layers, putting the main focus on development reported within the past three years but without omitting the fundamentals., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

We apply wide-field fluorescence microscopy to measure real-time attachment of photosynthetic proteins to plasmonically active silver nanowires. The observation of this effect is enabled, on the one hand, by sensitive detection of fluorescence and, on the other hand, by plasmonic enhancement of protein fluorescence. We examined two sample configurations with substrates being a bare glass coverslip and a coverslip functionalized with a monolayer of streptavidin. The different preparation of the substrate changes the observed behavior as far as attachment of the protein is concerned as well as its subsequent photobleaching. For the latter substrate the conjugation process is measurably slower. The described method can be universally applied in studying protein-nanostructure interactions for real-time fluorescence-based sensing., Competing Interests: The authors declare no conflict of interest.

The demand for electrochemical sensors with high sensitivity and reliability, fast response, and excellent selectivity has stimulated intensive research on developing highly active nanomaterials. In this work, freestanding 3D/Co 3 O 4 thorn-like and wire-like (nanowires) nanostructures are directly grown on a flexible carbon fiber paper (CFP) substrate by a single-step hydrothermal process without using surfactants or templates. The 3D/Co 3 O 4 thorn-like nanostructures show higher electrochemical activity than wire-like because of their high conductivity, large specific surface areas, and mesopores on their surface. The characterization of 3D/Co 3 O 4 nanostructures is performed by using high resolution transmission electron microscopy (HRTEM), field emission scanning electron microscopy (FESEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction analysis (XRD), and electrochemical methods. The 3D/Co 3 O 4 thorn-like nanostructures displayed non-enzymatic higher catalytic activity towards the electrochemical detection of glucose, compared to the 3D/Co 3 O 4 wire-like morphology. The 3D/Co 3 O 4 thorn-like nanostructures show a wide linear range response of glucose concentration ranging from 1 to 1000 μM with a detection limit of 0.046 μM (S/N = 3). The 3D/Co 3 O 4 thorn-like nanostructure-modified CFP electrode selectively detects glucose in the presence of 100-fold excess of interfering compounds. The 3D/Co 3 O 4 thorn-like nanostructure-modified CFP electrode is tested with human blood serum samples and validated with commercial glucose sensors. The newly developed sensor material shows potential for glucose monitoring in clinical and food samples.

Background: Th2 cytokines like interleukin-4, -5, and -13 are regarded as important drivers of the immunopathology underlying allergic rhinitis (AR) and asthma. The present study explores the capacity of pentosan polysulfate sodium (PPS), a semi-synthetic heparin-like macromolecular carbohydrate, to bind Th2 cytokines and exert biological neutralization in vitro, as well as anti-inflammatory actions in vivo., Methodology: The capacity of PPS to bind recombinant Th2 cytokines was tested with surface plasmon resonance (SPR) technology and biological Th2 neutralization was assessed by Th2-dependent proliferation assays. The in vivo anti-inflammatory action of PPS was studied using a validated Guinea-pig model of AR., Results: Binding studies revealed a strong and specific binding of PPS to IL-4, IL-5, and IL-13 with IC values suggesting as stronger cytokine binding than for heparin. Cytokine binding translated to a biological neutralization as PPS dose dependently inhibited Th2-dependent cell proliferation. Topical administration of PPS 30 min prior to nasal allergen challenge of sensitized animals significantly reduced late phase plasma extravasation, luminal influx of eosinophils, neutrophils, and total lavage leukocytes. Similar, albeit not statistically secured, effects were found for tissue leukocytes and mucus hyper-secretion. The anti-inflammatory effects of PPS compared favorably with established topical nasal steroid treatment., Conclusion: This study points out PPS as a potent Th2 cytokine-binding molecule with biological neutralization capacity and broad anti-inflammatory effects in vivo. As such PPS fulfills the role as a potential candidate molecule for the treatment of AR and further studies of clinical efficacy seems highly warranted., (© 2017 The Authors. Immunity, Inflammation and Disease Published by John Wiley & Sons Ltd.)

This work discusses an application of titanium oxide (TiO x ) thin films deposited using physical (reactive magnetron sputtering, RMS) and chemical (atomic layer deposition, ALD) vapour deposition methods as a functional coating for label-free optical biosensors. The films were applied as a coating for two types of sensors based on the localised surface plasmon resonance (LSPR) of gold nanoparticles deposited on a glass plate and on a long-period grating (LPG) induced in an optical fibre. Optical and structural properties of the TiO x thin films were investigated and discussed. It has been found that deposition method has a significant influence on optical properties and composition of the films, but negligible impact on TiO x surface silanization effectiveness. A higher content of oxygen with lower Ti content in the ALD films leads to the formation of layers with higher refractive index and slightly higher extinction coefficient than for the RMS TiO x . Moreover, application of the TiO x film independently on deposition method enables not only for tuning of the spectral response of the investigated biosensors, but also in case of LSPR for enhancing the ability for biofunctionalization, i.e., TiO x film mechanically protects the nanoparticles and induces change in the biofunctionalization procedure to the one typical for oxides. TiO x coated LSPR and LPG sensors with refractive index sensitivity of close to 30 and 3400nm/RIU, respectively, were investigated. The ability for molecular recognition was evaluated with the well-known complex formation between avidin and biotin as a model system. The shift in resonance wavelength reached 3 and 13.2nm in case of LSPR and LPG sensors, respectively. Any modification in TiO x properties resulting from the biofunctionalization process can be also clearly detected., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Robust detection of bacteria can significantly reduce risks of nosocomial infections, which are a serious problem even in developed countries (4.1 million cases each year in Europe). Here we demonstrate utilization of novel multifunctional bioconjugates as specific probes for bacteria detection. Bifunctional magnetic-fluorescent microparticles are coupled with bacteriophages. The T4 bacteriophage, due to its natural affinity to bacterial receptors, namely, OmpC and LPS, enables specific and efficient detection of Escherichia coli bacteria. Prepared probes are cheap, accessible (even in nonbiological laboratories), as well as versatile and easily tunable for different bacteria species. The magnetic properties of the bioconjugates facilitate the separation of captured target bacteria from other components of complex samples and other bacteria strains. Fluorescence enables simple analysis. We chose flow cytometry as the detection method as it is fast and widely used for biotests. The capture efficiency of the prepared bioconjugates is close to 100% in the range of bacteria concentrations from tens to around 10 5 CFU/mL. The limit of detection is restricted by flow cytometry capabilities and in our case was around 10 4 CFU/mL.

We show that surface-enhanced Raman spectroscopy (SERS) coupled with principal component analysis (PCA) can serve as a fast, reliable, and easy method for detection and identification of food-borne bacteria, namely Salmonella spp., Listeria monocytogenes, and Cronobacter spp., in different types of food matrices (salmon, eggs, powdered infant formula milk, mixed herbs, respectively). The main aim of this work was to introduce the SERS technique into three ISO (6579:2002; 11290-1:1996/A1:2004; 22964:2006) standard procedures required for detection of these bacteria in food. Our study demonstrates that the SERS technique is effective in distinguishing very closely related bacteria within a genus grown on solid and liquid media. The advantages of the proposed ISO-SERS method for bacteria identification include simplicity and reduced time of analysis, from almost 144 h required by standard methods to 48 h for the SERS-based approach. Additionally, PCA allows one to perform statistical classification of studied bacteria and to identify the spectrum of an unknown sample. Calculated first and second principal components (PC-1, PC-2) account for 96, 98, and 90% of total variance in the spectra and enable one to identify the Salmonella spp., L. monocytogenes, and Cronobacter spp., respectively. Moreover, the presented study demonstrates the excellent possibility for simultaneous detection of analyzed food-borne bacteria in one sample test (98% of PC-1 and PC-2) with a goal of splitting the data set into three separated clusters corresponding to the three studied bacteria species. The studies described in this paper suggest that SERS represents an alternative to standard microorganism diagnostic procedures. Graphical Abstract New approach of the SERS strategy for detection and identification of food-borne bacteria, namely S. enterica, L. monocytogenes, and C. sakazakii in selected food matrices., Competing Interests: The authors declare that they have no conflict of interests.

Uniform growth is a desirable trait in all large-scale animal production systems because it simplifies animal management and increases profitability. In parental broiler flocks, so-called broiler breeders, low growth uniformity is largely attributed to the feed competition that arises from quantitatively restricted feeding. As feed restriction is crucial to maintaining healthy and fertile breeders, several practices for reducing feed competition and the associated growth heterogeneity have been suggested and range from nutrient dilution by increasing fiber content in feed to intermittent fasting with increased portion size ("skip a day"), but no practice appears to be entirely effective. The fact that a large part of the heterogeneity remains even when feed competition is minimized suggests that some growth variation is caused by other factors. We investigated whether this variation arises during embryonic development (as measured by size at hatch) or during posthatch development by following the growth and body composition of birds of varying hatch sizes. Our results support the posthatch alternative, with animals that later grow to be small or large (here defined as >1 SD lighter or heavier than mean BW of the flock) being significantly different in size as early as 1 d after gaining access to feed ( < 0.05). We then investigated 2 possible causes for different postnatal growth: that high growth performance is linked 1) to interindividual variations in metabolism (as measured by cloacal temperature and verified by respirometry) or 2) to higher levels of social motivation (as measured in a social reinstatement T-maze), which should reduce the stress of being reared in large-scale commercial flocks. Neither of these follow-up hypotheses could account for the observed heterogeneity in growth. We suggest that the basis of growth heterogeneity in broiler breeder pullets may already be determined at the time of hatch in the form of qualitatively different maternal investments or immediately thereafter as an indirect result of differences in incubation conditions, hatching time, and resulting fasting time. Although this potential difference in maternal investment is not seen in body mass, tarsometatarsal length, or full body length of day-old chicks arriving at the farm, it may influence the development of differential feed and water intake during the first day of feeding, which in turn has direct effects on growth heterogeneity.

With the advent of nanotechnology, carbon nanomaterials such as carbon nanofibers (CNF) have aroused substantial interest in various research fields, including energy storage and sensing. Further improvement of their properties might be achieved via the application of viral particles such as bacteriophages. In this report, we present a filamentous M13 bacteriophage with a point mutation in gene VII (pVII-mutant-M13) that selectively binds to the carbon nanofibers to form 3D structures. The phage-display technique was utilized for the selection of the pVII-mutant-M13 phage from the phage display peptide library. The properties of this phage make it a prospective candidate for a scaffold material for CNFs. The results for binding of CNF by mutant phage were compared with those for maternal bacteriophage (pVII-M13). The efficiency of binding between pVII-mutant-M13 and CNF is about 2 orders of magnitude higher compared to that of the pVII-M13. Binding affinity between pVII-mutant-M13 and CNF was also characterized using atomic force microscopy, scanning electron microscopy, and transmission electron microscopy, which confirmed the specificity of the interaction of the phage pVII-mutant-M13 and the CNF; the binding occurs via the phage's ending, where the mutated pVII protein is located. No similar behavior has been observed for other carbon nanomaterials such as graphite, reduced graphene oxide, single-walled carbon nanotubes, and multiwalled carbon nanotubes. Infrared spectra confirmed differences in the interaction with CNF between the pVII-mutant-M13 and the pVII-M13. Basing on conducted research, we hypothesize that the interactions are noncovalent in nature, with π-π interactions playing the dominant role. Herein, the new bioconjugate material is introduced.

Background: An infection-immune association of periodontal disease with rheumatoid arthritis has been suggested. This study aimed to investigate the effect of pre-existing periodontitis on the development and the immune/inflammatory response of pristane-induced arthritis., Methods: We investigated the effect of periodontitis induced by ligature placement and Porphyromonas gingivalis (P. gingivalis) infection, in combination with Fusobacterium nucleatum to promote its colonization, on the development of pristane-induced arthritis (PIA) in rats (Dark Agouti). Disease progression and severity of periodontitis and arthritis was monitored using clinical assessment, micro-computed tomography (micro-CT)/intraoral radiographs, antibody response, the inflammatory markers such as α-1-acid glycoprotein (α-1-AGP) and c-reactive protein (CRP) as well as cytokine multiplex profiling at different time intervals after induction., Results: Experimentally induced periodontitis manifested clinically (P < 0.05) prior to pristane injection and progressed steadily until the end of experiments (15 weeks), as compared to the non-ligated arthritis group. Injection of pristane 8 weeks after periodontitis-induction led to severe arthritis in all rats demonstrating that the severity of arthritis was not affected by the pre-existence of periodontitis. Endpoint analysis showed that 89% of the periodontitis-affected animals were positive for antibodies against arginine gingipain B and furthermore, the plasma antibody levels to a citrullinated P. gingivalis peptidylarginine deiminase (PPAD) peptide (denoted CPP3) were significantly (P < 0.05) higher in periodontitis rats with PIA. Additionally, there was a trend towards increased pro-inflammatory and anti-inflammatory cytokine levels, and increased α-1-AGP levels in plasma from periodontitis-challenged PIA rats., Conclusions: Pre-existence of periodontitis induced antibodies against citrullinated peptide derived from PPAD in rats with PIA. However, there were no differences in the development or severity of PIA between periodontitis challenged and periodontitis free rats.

Treatment of high-risk childhood neuroblastoma is a clinical challenge which has been hampered by a lack of reliable neuroblastoma mouse models for preclinical drug testing. We have previously established invasive and metastasising patient-derived orthotopic xenografts (PDXs) from high-risk neuroblastomas that retained the genotypes and phenotypes of patient tumours. Given the important role of the tumour microenvironment in tumour progression, metastasis, and treatment responses, here we analysed the tumour microenvironment of five neuroblastoma PDXs in detail. The PDXs resembled their parent tumours and retained important stromal hallmarks of aggressive lesions including rich blood and lymphatic vascularisation, pericyte coverage, high numbers of cancer-associated fibroblasts, tumour-associated macrophages, and extracellular matrix components. Patient-derived tumour endothelial cells occasionally formed blood vessels in PDXs; however, tumour stroma was, overall, of murine origin. Lymphoid cells and lymphatic endothelial cells were found in athymic nude mice but not in NSG mice; thus, the choice of mouse strain dictates tumour microenvironmental components. The murine tumour microenvironment of orthotopic neuroblastoma PDXs reflects important hallmarks of aggressive and metastatic clinical neuroblastomas. Neuroblastoma PDXs are clinically relevant models for preclinical drug testing., (Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)

All aerobic organisms are to some degree affected by oxidative stress, which is an imbalance between pro-oxidants and antioxidants in favor of the former. Pro-oxidants can damage DNA, proteins and lipids, and as such oxidative stress can carry considerably fitness costs. In mammals excessive calorie intake is a known cause of oxidative stress. We investigated whether in migrant birds, which typically engage in over-eating in between flights (refueling), high food intake causes oxidative stress. In an experiment we compared levels of plasmatic total non-enzymatic antioxidant capacity (AOX) and oxidative damage (lipid peroxidation) between migrants repeatedly fasted and refed (simulating the flight-refuel cycle of wild migrants), and migrants on ad libitum food. We found that refueling increased AOX, an effect mainly attributable to an increase in uric acid level, an antioxidant that is produced during protein metabolism. Accordingly, variation in AOX was mainly explained by the refueling birds' food intake. However, food intake in migrants on ad libitum food did not explain any variation in AOX. Refueling did not affect lipid peroxidation, nor were its levels explained by food intake. We propose that over-eating migrants retain uric acid, which might be a very low cost mechanism to forego oxidative damage., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Single molecule surface-enhanced resonance Raman scattering (SERRS) spectra have been obtained for the parent porphycene (Pc-d0) and its deuterated isotopologue (Pc-d12), located on gold and silver nanoparticles. Equal populations of "hot spots" by the two isotopologues are observed for 1 : 1 mixtures in a higher concentration range of the single molecule regime (5 × 10(-9) M). For decreasing concentrations, hot spots are preferentially populated by undeuterated molecules. This is interpreted as an indication of a lower surface diffusion coefficient of Pc-d12. The photostability of single Pc molecules placed on nanoparticles is strongly increased in comparison with polymer environments. Trans tautomeric species dominate the spectra, but the analysis of time traces reveals transient intermediates, possibly due to rare cis tautomeric forms.

Three-dimensional nickel-iron (3-D/Ni-Fe) nanostructures are exciting candidates for various applications because they produce more reaction-active sites than 1-D and 2-D nanostructured materials and exhibit attractive optical, electrical and catalytic properties. In this work, freestanding 3-D/Ni-Fe interconnected hierarchical nanosheets, hierarchical nanospheres, and porous nanospheres are directly grown on a flexible carbon fiber paper (CFP) substrate by a single-step hydrothermal process. Among the nanostructures, 3-D/Ni-Fe interconnected hierarchical nanosheets show excellent electrochemical properties because of its high conductivity, large specific active surface area, and mesopores on its walls (vide infra). The 3-D/Ni-Fe hierarchical nanosheet array modified CFP substrate is further explored as a novel electrode for electrochemical non-enzymatic glucose sensor application. The 3-D/Ni-Fe hierarchical nanosheet arrays exhibit significant catalytic activity towards the electrochemical oxidation of glucose, as compared to the 3-D/Ni-Fe hierarchical nanospheres, and porous nanospheres. The 3-D/Ni-Fe hierarchical nanosheet arrays can access a large amount of glucose molecules on their surface (mesopore walls) for an efficient electrocatalytic oxidation process. Moreover, 3-D/Ni-Fe hierarchical nanosheet arrays showed higher sensitivity (7.90 μA μM(-1) cm(-2)) with wide linear glucose concentration ranging from 0.05 μM to 0.2 mM, and the low detection limit (LOD) of 0.031 μM (S/N = 3) is achieved by the amperometry method. Further, the 3-D/Ni-Fe hierarchical nanosheet array modified CFP electrode can be demonstrated to have excellent selectivity towards the detection of glucose in the presence of 500-fold excess of major important interferents. All these results indicate that 3-D/Ni-Fe hierarchical nanosheet arrays are promising candidates for non-enzymatic glucose sensing.

Syntheses of [Co(bpy)3](2+) yield racemic solutions because the Δ- and Λ-enantiomers are stereochemically labile. However, crystallization and attrition-enhanced deracemization can give homochiral crystal batches of either handedness in quantitative yield. Subsequently, solvent-free oxidation with bromine vapour fixes the chirality because [Co(bipy)3](3+) does not enantiomerize in solution at ambient temperature. This combination of Viedma ripening and the labile/inert Co(II)/Co(III) couple constitutes a convenient method of absolute asymmetric synthesis., (© 2015 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.)

Supported tungsten carbide is an efficient and vital nanomaterial for the development of high-performance, sensitive, and selective electrochemical sensors. In this work, tungsten carbide with tube-like nanostructures (WC NTs) supported platinum nanoparticles (PtNPs) are synthesized and explored as an efficient catalyst toward electrochemical oxidation of oxalic acid for the first the time. The WC NTs supported PtNPs modified glassy carbon (GC) electrode is highly sensitive toward the electrochemical oxidation of oxalic acid. A large decrease in the oxidation overpotential (220 mV) and significant enhancement in the peak current compared to unmodified and Pt/C modified GC electrodes have been observed without using any redox mediator. Moreover, WC NTs supported PtNPs modified electrode possessed wide linear concentration ranges from 0 to 125 nM and a higher sensitivity toward the oxidation of oxalic acid (80 nA/nM) achieved by the amperometry method. The present modified electrode showed an experimentally determined lowest detection limit (LOD) of 12 nM (S/N = 3). Further, WC NTs supported PtNPs electrode can be demonstrated to have an excellent selectivity toward the detection of oxalic acid in the presence of a 200-fold excess of major important interferents. The practical application of WC NTs supported PtNPs has also been demonstrated in the detection of oxalic acid in tomato fruit sample, by differential pulse voltammetry under optimized conditions.