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Publisher Copyright: © 2021 The Authors. Published by American Chemical Society. Although extractives have been symbolized as major bioactive pharmacological compounds from Salix (Salicaceae) bark, we speculated that these pharmaceutical effects cannot be solely attributed to phenolic components and their derivatives, but the long-chain suberin acids also contribute to their therapeutic effects. Hence, isolation and deconstruction of suberin were conducted, for the first time, to enrich our knowledge about the macromolecular components at the cell wall of willow bark. Saponification was adopted to obtain suberin extracts at a yield of approximately 5 wt % based on the bark of the studied hybrids. Gas chromatography-mass spectrometry allowed qualification and quantification of 23 compounds from the released suberin monomers, from which fatty acids represented majority of the isolated suberin, namely, fatty acid methyl esters (C17-C19); mono-carboxylic acid (C7-C16); alpha, omega-dicarboxylic acid (C7-C16); and omega-hydroxy long-chain fatty acids (C16-C22). Additionally, the lipophilic extractive was dominated by piceol, heptacosane, β-sitosterol, and fatty acids (C16-C28) from the studied hybrids. These findings could boost our integrative approach toward full valorization of willow bark.

Background Primary diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is based on detection of virus RNA in nasopharyngeal swab samples. In addition, analysis of humoral immunity against SARS-CoV-2 has an important role in viral diagnostics and seroprevalence estimates. Methods We developed and optimized an enzyme immunoassays (EIA) using SARS-CoV-2 nucleoprotein (N), S1 and receptor binding domain (RBD) of the viral spike protein, and N proteins from SARS, Middle East respiratory syndrome (MERS), and 4 low-pathogenic human CoVs. Neutralizing antibody activity was compared with SARS-CoV-2 IgG, IgA, and IgM EIA results. Results The sensitivity of EIA for detecting immune response in COVID-19 patients (n = 101) was 77% in the acute phase and 100% in the convalescent phase of SARS-CoV-2 infection when N and RBD were used as antigens in IgG and IgA specific EIAs. SARS-CoV-2 infection significantly increased humoral immune responses against the 229E and NL63 N proteins. S1 and RBD-based EIA results had a strong correlation with microneutralization test results. Conclusions The data indicate a combination of SARS-CoV-2 S1 or RBD and N proteins and analysis of IgG and IgA immunoglobulin classes in sera provide an excellent basis for specific and sensitive serological diagnostics of COVID-19.

Recent studies suggest that professional athletes seem to experience significant sleeping problems. However, little is still known about the occurrence of sleeping challenges at different stages of an athletic career. This descriptive study aimed to compare the sleep of professional athletes with younger elite amateur athletes. A total of 401 sportsmen, 173 youth elite amateur athletes and 228 professional athletes fulfilled a validated questionnaire. The self-estimated quality of sleep (on a linear scale 0–10) was significantly better in youth, being 7.9 compared to 7.4 (p p p

In Arabidopsis, two leaf-type ferredoxin-NADP(+) oxidoreductase (LFNR) isoforms function in photosynthetic electron flow in reduction of NADP(+), while two root-type FNR (RFNR) isoforms catalyse reduction of ferredoxin in non-photosynthetic plastids. As the key to understanding, the function of RFNRs might lie in their spatial and temporal distribution in different plant tissues and cell types, we examined expression of RFNR1 and RFNR2 genes using beta-glucuronidase (GUS) reporter lines and investigated accumulation of distinct RFNR isoforms using a GFP approach and Western blotting upon various stresses. We show that while RFNR1 promoter is active in leaf veins, root tips and in the stele of roots, RFNR2 promoter activity is present in leaf tips and root stele, epidermis and cortex. RFNR1 protein accumulates as a soluble protein within the plastids of root stele cells, while RFNR2 is mainly present in the outer root layers. Ozone treatment of plants enhanced accumulation of RFNR1, whereas low temperature treatment specifically affected RFNR2 accumulation in roots. We further discuss the physiological roles of RFNR1 and RFNR2 based on characterization of rfnr1 and rfnr2 knock-out plants and show that although the function of these proteins is partly redundant, the RFNR proteins are essential for plant development and survival.

Salmonines in the Baltic Sea and North American lakes suffer from thiamine (vitamin B1) deficiency, which is connected to an abundant lipid-rich diet containing substantial amounts of polyunsaturated fatty acids (PUFAs). In the Baltic region, this is known as the M74 syndrome. It affects both adult salmon (Salmo salar) and especially their offspring, impairing recruitment. However, very little is known about the thiamine and lipid metabolism of salmon during feeding and spawning migrations in the Baltic Sea. In this study, salmon females were sampled along the spawning run from the southern Baltic Proper in four locations at sea and finally at spawning in a river at the Bothnian Bay in a year with insignificant M74 mortality. Changes in concentrations of thiamine and its components in muscle, ovaries, and the liver and other biochemical indices potentially relating to lipid and fatty acid metabolism were investigated. The results provide further evidence of the role of peroxidation of PUFAs in eliciting thiamine deficiency in salmon: During the entire spawning run, the muscle total lipid content decreased by 50%, palmitic acid (16:0) by 62%, and docosahexaenoic acid (DHA, 22:6n-3) by 45%. The concentration of total thiamine decreased significantly until the spawning in the liver and ovaries, 66 and 70% respectively. In the muscle, the proportion of thiamine pyrophosphate of total thiamine increased with the use of muscular lipid stores. There was no trend in the concentration of total carotenoids during the spawning run. The doubling of the concentration of hepatic malondialdehyde indicated peroxidation of PUFAs, and the mobilisation of body lipids suppressed the activity of glucose-6-phosphate dehydrogenase, as consumed dietary lipids would also have done.

Background B cells play a crucial role during rhinovirus (RV) infections by production of virus‐neutralizing antibodies. A main feature of common variable immunodeficiency (CVID) is hypogammaglobulinemia (HG). HG patients have severely reduced levels of antibody‐producing B cells and suffer from prolonged virus infections. Here, we addressed whether antiviral response of peripheral blood lymphocytes differs between HG patients and healthy individuals during natural RV infection. Methods Using fluorescence‐activated cell sorting, B‐cell subsets were analyzed. Simultaneously, CD19 + B cells, CD14 + monocytes, and CD3 + T cells were sorted from frozen peripheral blood mononuclear cells from 11 RV‐infected hypogammaglobulinemia patients, 7 RV‐infected control subjects, and 14 noninfected control subjects. Real‐time PCR was used to study expression of antiviral genes. A pan‐RV PCR was used to detect RV genome in all samples. Results In HG patients, total B‐cell numbers, as well as IgA + and IgG + switched memory B cells, were reduced while naive B cells and T cells were increased. STAT1 expression was increased in HG patients compared to controls in all lymphocyte subsets analyzed. The expression of antiviral genes IFITM1 and MX1 correlated with STAT1 expression in B cells and monocytes. RV RNA was found in 88.9% of monocytes from infected HG patients, 85.7% of monocytes from infected controls, and 7.1% of monocytes from uninfected controls. Conclusions We demonstrate an increased antiviral response in B cells and monocytes in HG patients and their correlation with STAT1 expression. Monocytes of infected HG patients and infected non‐HG controls carry RV RNA.

Polymerase chain reaction (PCR) detection has become the gold standard for diagnosis and typing of enterovirus (EV) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using the appropriate sample types and high assay sensitivity as viral loads in cerebrospinal fluid samples from meningitis and sepsis clinical presentation can be extremely low. This study evaluated the sensitivity and specificity of currently used commercial and in‐house diagnostic and typing assays. Accurately quantified RNA transcript controls were distributed to 27 diagnostic and 12 reference laboratories in 17 European countries for blinded testing. Transcripts represented the four human EV species (EV‐A71, echovirus 30, coxsackie A virus 21, and EV‐D68), HPeV3, and specificity controls. Reported results from 48 in‐house and 15 commercial assays showed 98% detection frequencies of high copy (1000 RNA copies/5 µL) transcripts. In‐house assays showed significantly greater detection frequencies of the low copy (10 copies/5 µL) EV and HPeV transcripts (81% and 86%, respectively) compared with commercial assays (56%, 50%; P = 7 × 10−5). EV‐specific PCRs showed low cross‐reactivity with human rhinovirus C (3 of 42 tests) and infrequent positivity in the negative control (2 of 63 tests). Most or all high copy EV and HPeV controls were successfully typed (88%, 100%) by reference laboratories, but showed reduced effectiveness for low copy controls (41%, 67%). Stabilized RNA transcripts provide an effective, logistically simple and inexpensive reagent for evaluation of diagnostic assay performance. The study provides reassurance of the performance of the many in‐house assay formats used across Europe. However, it identified often substantially reduced sensitivities of commercial assays often used as point‐of‐care tests. © 2019 The Authors. Journal of Medical Virology published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

This first pilot trial on external quality assessment (EQA) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) whole-genome sequencing, initiated by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) Study Group for Genomic and Molecular Diagnostics (ESGMD) and the Swiss Society for Microbiology (SSM), aims to build a framework between laboratories in order to improve pathogen surveillance sequencing. Ten samples with various viral loads were sent out to 15 clinical laboratories that had free choice of sequencing methods and bioinformatic analyses. The key aspects on which the individual centers were compared were the identification of (i) single nucleotide polymorphisms (SNPs) and indels, (ii) Pango lineages, and (iii) clusters between samples. The participating laboratories used a wide array of methods and analysis pipelines. Most were able to generate whole genomes for all samples. Genomes were sequenced to various depths (up to a 100-fold difference across centers). There was a very good consensus regarding the majority of reporting criteria, but there were a few discrepancies in lineage and cluster assignments. Additionally, there were inconsistencies in variant calling. The main reasons for discrepancies were missing data, bioinformatic choices, and interpretation of data. The pilot EQA was overall a success. It was able to show the high quality of participating laboratories and provide valuable feedback in cases where problems occurred, thereby improving the sequencing setup of laboratories. A larger follow-up EQA should, however, improve on defining the variables and format of the report. Additionally, contamination and/or minority variants should be a further aspect of assessment.

Objective Cytokine release syndrome is suggested to be the most important mechanism triggering acute respiratory distress syndrome and end organ damage in COVID-19. The severity of disease may be measured by different biomarkers. Methods We studied markers of inflammation and coagulation as recorded in 29 patients on admission to the hospital in order to identify markers of severe COVID-19 and need of ICU. Results Patients who were eventually admitted to ICU displayed significantly higher serum levels of interleukin-6 (IL-6), C-reactive protein (CRP), and procalcitonin. No statistical differences were found between the groups in median levels of lymphocytes, D-dimer or ferritin. Conclusions IL-6 and CRP were the strongest predictors of severity in hospitalized patients with COVID-19.

The majority of asthma exacerbations in children are caused by Rhinovirus (RV), a positive sense single stranded RNA virus of the Picornavirus family. The host has developed virus defense mechanisms that are mediated by the upregulation of interferon-activated signaling. However, the virus evades the immune system by inducing immunosuppressive cytokines and surface molecules like programmed cell death protein 1 (PD-1) and its ligand (PD-L1) on immunocompetent cells. Initially, RV infects epithelial cells, which constitute a physiologic mucosal barrier. Upon virus entrance, the host cell immediately recognizes viral components like dsRNA, ssRNA, viral glycoproteins or CpG-DNA by host pattern recognition receptors (PRRs). Activation of toll like receptors (TLR) 3, 7 and 8 within the endosome and through MDA-5 and RIG-I in the cytosol leads to the production of interferon (IFN) type I and other antiviral agents. Every cell type expresses IFNAR1/IFNAR2 receptors thus allowing a generalized antiviral activity of IFN type I resulting in the inhibition of viral replication in infected cells and preventing viral spread to non-infected cells. Among immune evasion mechanisms of the virus, there is downregulation of IFN type I and its receptor as well as induction of the immunosuppressive cytokine TGF-β. TGF-β promotes viral replication and is associated with induction of the immunosuppression signature markers LAP3, IDO and PD-L1. This article reviews the recent advances on the regulation of interferon type I expression in association with RV infection in asthmatics and the immunosuppression induced by the virus.

Funding Information: This work made use of the Aalto University Nanomicroscopy Center (Aalto-NMC) premises. The authors thank Rita Hatakka from Aalto University for her skillful assistance in HPAEC measurement. This work was a part of the Academy of Finland's Flagship Programme under Projects No. 318890 and 318891 (Competence Center for Materials Bioeconomy, FinnCERES). Authors thank RAIZ (Portuguese Research Institute on Forestry and Paper) for supplying the eucalyptus bark sample used within the scope of the project Inpactus – innovative products and technologies from eucalyptus, Project N.º 21874 funded by Portugal 2020 through European Regional Development Fund (ERDF) in the frame of COMPETE 2020 nº246/AXIS II/2017. Publisher Copyright: © 2021 The Author(s) Copyright: Copyright 2021 Elsevier B.V., All rights reserved. High surface lignin content (i.e. 34.6 %) sclerenchyma fiber bundles were successfully isolated with a yield of 71 % by a mild alkali (NaOH dosage of 5 wt%) extraction of eucalyptus (Eucalyptus globulus) bark under 100 °C for 60 min. The mechanical properties of the composites prepared by hot pressing of cellulose acetate butyrate (CAB) sheets with the fiber bundles were evaluated. The fiber bundles exhibited good compatibility with CAB due to their hydrophobic fiber surfaces. The mechanical properties of the fiber bundle/ cellulose acetate butyrate composites revealed the maximum at a weight ratio of 25:75, which demonstrates the promise of utilizing these isotropic aligned fiber bundles as the reinforcement to the cellulose acetate butyrate without the addition of plasticizers for composite uses.

Highlights • Fish caught in the Baltic Sea and Finland's freshwaters were analysed for 13 PFAAs. • ∑PFAA concentrations were highest in the Baltic smelt. • PFOS was most frequently detected PFAA in fish from freshwaters and the Baltic Sea. • PFOS concentration in all species except in smelt was below the EQSbiota. • TWI derived by EFSA for ∑PFAS-4 was exceeded in 10 of 13 Baltic fish species. Occurrence and distribution of perfluoroalkyl acids (PFAAs), a sub-category of per- and polyfluoroalkyl substances (PFASs), is widespread in the environment. Food, especially fish meat, is a major pathway via which humans are exposed to PFAAs. As fish is an integral part of Nordic diet, therefore, in this study, several fish species, caught in selected Baltic Sea basins and freshwater bodies of Finland, were analysed for PFAAs. Perfluorooctane sulfonate (PFOS) was detected in all Baltic Sea fish samples and in >80% fish samples from freshwaters. PFOS contributed between 46 and 100% to the total PFAA concentration in Baltic Sea fish samples and between 19 and 28% in fish samples from freshwaters. Geographically, concentration ratios of PFOS to other PFAAs differed between fish from the Baltic Sea and Finnish lakes suggesting that distribution of PFAAs differ in these environments. Results were compared with current safety thresholds – environmental quality standard for biota (EQSbiota) set by the European Commission and a group tolerable weekly intake (TWI) for the sum of four PFASs (∑PFAS-4) i.e. perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorohexane sulfonate (PFHxS) and PFOS, recommended by the European Food Authority (EFSA). EQSbiota compliance was observed for PFOS in all species except smelt caught in the Baltic Sea and also in the River Aurajoki, where smelt had migrated from the Baltic Sea for spawning. Moderate consumption of most Baltic fishes (200 g week−1) results in an exceedance of the new TWI (4.4 ng kg−1 body weight week−1) for ∑PFAS-4.

Acute Myeloid Leukaemia (AML) continues to have a poor prognosis, especially in the elderly. One reason for this is that many treatment regimens are not well tolerated by elderly patients. Much current focus is on the development of therapies that can target specific vulnerabilities of AML while having fewer toxic side effects. However, despite much recent progress in developing better drugs, many patients with AML still die within a year of diagnosis, partly due to the fact that it is difficult to identify therapeutic targets that are effective across multiple AML subtypes. One common factor across AML subtypes is the presence of a block in differentiation. Thus screening for compounds that can overcome this block in genetically diverse AML models should allow for the identification of agents that are not dependent on a specific mutation for their efficacy. Here, we used a phenotypic screen to identify novel compounds that stimulate differentiation in several AML cell lines. Lead compounds were shown to decrease tumour burden and to increase survival in vivo. Using multiple complementary target deconvolution approaches, these compounds were revealed to be anti-mitotic tubulin disruptors that cause differentiation by inducing a G2-M mitotic arrest. Together, these results reveal a novel function for tubulin disruptors in causing differentiation of AML cells.

The development of a mild and green method for separating natural fiber bundles from willow bark is an essential step in exploring and preserving their natural functions. The isolation of well-oriented fiber bundles from the bark of a fast-growing willow hybrid solely using sodium bicarbonate under mild conditions was successfully demonstrated. Additionally, Lyocell fibers were mixed with an equal amount of the willow bark fiber bundles and proved their ability to convert into spun yarns, which provided excellent protection for ultraviolet radiation (UPF ≥ 140). Moreover, these yarns demonstrated strong antibacterial activity (A ≥ 8) against the Gram-positive pathogen Staphylococcus aureus, resulting in complete eradication of viable bacteria after 24 -h incubation with the material. A laundering treatment had no effect on the UV protection or the antibacterial performance. Utilizing these inherent properties from natural fibers for technical textile applications is very promising.

Enzymatic modification of bleached softwood kraft fibres for improved fibre reactivity was studied at high (20% w/w) and low (1% w/w) dry matter content. The role of enzyme family and structure in fibre modification was assessed using endoglucanases from three structurally different glycoside hydrolase (GH) families (5, 7 and 45) with and without a carbohydrate binding module (CBM). Based on the amount of dissolved sugars, enzyme action at high consistency was about sixfold higher compared to a fibre treatment at low consistency. The GH45 endoglucanase was found to be most specific in acting on pulp cellulose whereas the family 5 and 7 endoglucanases had activity also on pulp hemicelluloses. The GH45 endoglucanase was found to be most efficient in reducing molecular weight and viscosity of the pulp. In addition, treatment with the GH45 endoglucanase resulted in the highest micropore volume in fibres and thus an increase in cellulose accessibility. The increased accessibility could be seen as decreased dissolution time in cupriethylenediamine using recently developed analytical techniques: viscometric analysis and microscopic video analysis. At high consistency, CBM was not promoting enzyme action, although CBMs are known to be beneficial at low dry matter conditions. Graphical abstract: [Figure not available: see fulltext.].

Global climate change can affect the energy content of fish by altering their lipid physiology and consumption. We investigated the effects of different environmental stressors on the lipid content of the Baltic herring (Clupea harengus membras) from spawning ground samples that were collected annually in the northern Baltic Sea. During 1987–2014, the average lipid content of herring muscle decreased from 5%–6% (wet mass) to 1.5% (wet mass). Generalized linear mixed models indicated that sea water salinity and the size of the herring stock explained best the declining trend of lipid content. We estimated that the amount of the lipid storage incorporated in the spawning stock decreased by approximately 45% during the study, with respective energy content decreases. Fatty acid composition analysis revealed that herring lipids contained a high proportion of EPA (eicosapentaenoic acid; 20:5n-3) and DHA (docosahexaenoic acid; 22:6n-3), which likely originated from its main summertime prey, Limnocalanus macrurus. The results illustrate various climate change-induced processes leading to changes in the lipid content of the Baltic herring and, consequently, to changes in the energy flows of the northern Baltic ecosystem.

The bark from fast-growing willow crops contains long and strong sclerenchyma fiber bundles, which could potentially replace the pulp and annual plant fibers currently used to reinforce green polymer composites. Here we successfully demonstrate the isolation of fiber bundles of willow bark with a simple alkali treatment under much milder conditions than what pulp fiber separation requires. The fiber bundles separated had hydrophobic surfaces, which made them compatible with polymers, in this case with polylactic acid, without using any additives. The most hydrophobic fiber bundles of willow bark provided the strongest and toughest composites, superior to the corresponding isotropic composites of pulp and flax fibers. Integration of the fiber bundle isolation with a prior recovery of hot water extractable aromatics from the bark and further processing of the debarked willow stems into bioethanol and lignin, for example, could make the full valorization of the willow biomass feasible.

Prostate cancer is heterogeneous and patients would benefit from methods that stratify those who are likely to respond to systemic therapy. We employed single-cell assays for transposase-accessible chromatin (ATAC) and RNA sequencing in models of early treatment response and resistance to enzalutamide. In doing so, we identified pre-existing and treatment-persistent cell subpopulations that possess transcriptional stem-like features and regenerative potential when subjected to treatment. We found distinct chromatin landscapes associated with enzalutamide treatment and resistance that were linked to alternative transcriptional programs. Transcriptional profiles characteristic of persistent stem-like cells were able to stratify the treatment response of patients. Ultimately, we show that defining changes in chromatin and gene expression in single-cell populations from pre-clinical models can reveal hitherto unrecognized molecular predictors of treatment response. This suggests that the high analytical resolution of pre-clinical models enabled by single-cell methods may powerfully inform clinical decision-making.

Overexploitation of natural resources is often viewed as a problem characteristic of only the human species. However, any species could evolve a capacity to overexploit its essential resources through natural selection and competition, even to the point of resource collapse. Here, we describe the processes that potentially lead to overexploitation and synthesize what is known about overexploitation limiters in other species. We propose that there are five pathways that counteract the evolutionary drive towards overexploitation and/or mitigate its consequences: top–down trophic control, interference, cost-efficiency tradeoffs, resource trait evolution, and spatial heterogeneity. These mechanisms constrain the number of exploiters and/or lower the rate of the resource usage at the individual level. We hypothesize that in ecosystems with reasonable functional diversity, coevolution strengthens this limiter network, preventing overexploitation, and thus argue that diversity begets stability via evolution. Violent population cycles in species-poor northern ecosystems and eruptions of invading alien species are exceptions that confirm this rule, because these ecosystems either lack functional diversity or there has not been enough time for coevolution to play out its stabilizing role. We propose that the overexploitation by our own species could be prevented via a network of socio-economical limiters that act in an analogous way.

Plants require interaction between signaling pathways to differentiate and integrate stress responses and deploy appropriate defenses. The hormones ethylene, salicylic acid (SA), and jasmonic acid (JA) are important regulators of plant defenses. Numerous interactions between these signaling pathways are the cornerstone of robust plant immunity. Additionally, during the early response to pathogens, reactive oxygen species (ROS) act as signaling molecules. Here, we examined the extent of signal interaction in the early stages of Botrytis cinerea infection. To enable a comparison between B. cinerea infection with ROS signaling, we subjected plants to ozone treatment, which stimulates an apoplastic ROS burst. We used a collection of single, double, and triple signaling mutants defective in hormone signaling and biosynthesis and subjected them to B. cinerea infection and ozone treatment at different timepoints. We examined lesion size, cell death, and gene expression (both quantitatively and spatially). The two treatments shared many similarities, especially in JA-insensitive mutants, which were sensitive to both treatments. Unexpectedly, a B. cinerea–susceptible JA-insensitive mutant (coi1), became tolerant when both SA biosynthesis and signaling was impaired (coi1 npr1 sid2), demonstrating that JA responses may be under the control of SA. Extensive marker gene analysis indicated JA as the main regulator of both B. cinerea and ozone defenses. In addition, we identified the transcription factor SR1 as a crucial regulator of PLANT DEFENSIN expression and cell-death regulation, which contributes to resistance to B. cinerea. Overall, our work further defines the context of ROS in plant defense signaling. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Thiamine (vitamin B1) deficiency of salmonines, caused by an abundant lipid-rich fish diet and consequently, the abundance of polyunsaturated fatty acids, is called the M74 syndrome in the Baltic Sea. Because of its deleterious effects on wild Atlantic salmon (Salmo salar) stocks and progeny production in fish cultivation, a model was developed to derive the annual female-specific mortality percentages of yolk-sac fry (YSFM) from the free thiamine concentrations of unfertilized eggs. In years with a high M74 incidence, thiamine-deficient females were larger, with a larger condition factor (CF) than non-M74 females. Otherwise, M74 females were generally smaller. The mean CF of M74 females was in most years higher than that of non-M74 females. The model compiled enables the cost-effective estimation of YSFM of individual female salmon, without the incubation of eggs and hatched yolk-sac fry for several months, thus benefitting the management of salmon stocks and their efficient utilization.

Duchenne muscular dystrophy is a fatal disease with no cure, caused by lack of the cytoskeletal protein dystrophin. Upregulation of utrophin, a dystrophin paralogue, offers a potential therapy independent of mutation type. The failure of first-in-class utrophin modulator ezutromid/SMT C1100 in Phase II clinical trials necessitates development of compounds with better efficacy, physicochemical and ADME properties and/or complementary mechanisms. We have discovered and performed a preliminary optimisation of a novel class of utrophin modulators using an improved phenotypic screen, where reporter expression is derived from the full genomic context of the utrophin promoter. We further demonstrate through target deconvolution studies, including expression analysis and chemical proteomics, that this compound series operates via a novel mechanism of action, distinct from that of ezutromid.

Asthma development and persistence is tightly linked to respiratory viruses. Viral presence is usually interrogated with targeted approaches during periods of disease activity and/or infections, thus neglecting viral occurrence during steady states. We investigate the virome in the upper respiratory system of healthy and asthmatic preschool children during asymptomatic/non-infection periods using metagenomics. Children with asthma have a characteristically dysbiotic virome that correlates to disease severity and control. The major component of dysbiosis is bacteriophage deficiency, while eukaryotic viral presence is increased. At the metacommunity level, differential virus species co-occurrence patterns suggest a decrease of the microbiota community resilience in asthma. Viral dysbiosis is therefore a key characteristic of asthma pathophysiology.

By adulthood, almost all humans become infected by at least one herpesvirus (HHV). The maladies inflicted by these microbes extend beyond the initial infection, as they remain inside our cells for life and can reactivate, causing severe diseases. The diagnosis of active infection by these ubiquitous pathogens includes the detection of DNA with sensitive and specific assays. We developed the first quantitative PCR assay (HERQ-9) designed to identify and quantify each of the nine human herpesviruses. The simultaneous detection of HHVs in the same sample is important since they may act together to induce life-threatening conditions. Moreover, the high sensitivity of our method is of extreme value for assessment of the effects of these viruses persisting in our body and their long-term consequences on our health., Infections with the nine human herpesviruses (HHVs) are globally prevalent and characterized by lifelong persistence. Reactivations can potentially manifest as life-threatening conditions for which the demonstration of viral DNA is essential. In the present study, we developed HERQ-9, a pan-HHV quantitative PCR designed in triplex reactions to differentiate and quantify each of the HHV-DNAs: (i) herpes simplex viruses 1 and 2 and varicella-zoster virus; (ii) Epstein-Barr virus, human cytomegalovirus, and Kaposi’s sarcoma-associated herpesvirus; and (iii) HHV-6A, -6B, and -7. The method was validated with prequantified reference standards as well as with mucocutaneous swabs and cerebrospinal fluid, plasma, and tonsillar tissue samples. Our findings highlight the value of multiplexing in the diagnosis of many unsuspected, yet clinically relevant, herpesviruses. In addition, we report here frequent HHV-DNA co-occurrences in clinical samples, including some previously unknown. HERQ-9 exhibited high specificity and sensitivity (LOD95s of ∼10 to ∼17 copies/reaction), with a dynamic range of 101 to 106 copies/μl. Moreover, it performed accurately in the coamplification of both high- and low-abundance targets in the same reaction. In conclusion, we demonstrated that HERQ-9 is suitable for the diagnosis of a plethora of herpesvirus-related diseases. Besides its significance to clinical management, the method is valuable for the assessment of hitherto-unexplored synergistic effects of herpesvirus coinfections. Furthermore, its high sensitivity enables studies on the human virome, often dealing with minute quantities of persisting HHVs. IMPORTANCE By adulthood, almost all humans become infected by at least one herpesvirus (HHV). The maladies inflicted by these microbes extend beyond the initial infection, as they remain inside our cells for life and can reactivate, causing severe diseases. The diagnosis of active infection by these ubiquitous pathogens includes the detection of DNA with sensitive and specific assays. We developed the first quantitative PCR assay (HERQ-9) designed to identify and quantify each of the nine human herpesviruses. The simultaneous detection of HHVs in the same sample is important since they may act together to induce life-threatening conditions. Moreover, the high sensitivity of our method is of extreme value for assessment of the effects of these viruses persisting in our body and their long-term consequences on our health.

Palatine tonsils have been observed to harbor several distinct respiratory and herpesviruses in separate studies. In this study, the presence of these viruses in palatine tonsils was comprehensively studied in both children and adults. A cross‐sectional analysis of 181 patients (median age 22 years; range, 2.6‐66) operated for a benign tonsillar disease was conducted. Real‐time polymerase chain reaction was performed to detect 27 distinct viruses in all: eight human herpesviruses, 16 respiratory viruses, parvo B19, and polyoma BK/JC viruses. Clinical characteristics of the patients and underlying conditions were evaluated. In total, 92% of patients had virus detected in tonsils (Epstein‐Barr virus 72%, human herpesvirus 7, and 6B 54% and 16%, respectively, enterovirus 18%, parvovirus B19 7% and the rest, Highlights This study comprehensively reports a cross‐sectional view of intratonsillar virus infections in 181 elective tonsillectomy patients (median age 22 years, range 2.6–66). Enterovirus was more common in children and was frequently observed in the presence of HHV6B. Tonsils are a major virus reservoir for distinct herpes and respiratory viruses without a positive association with tonsillar disease or respiratory symptoms.

Vegetation at ecotone transitions between open and forested areas is often heavily affected by two key processes: climate change and management of large herbivore densities. These both drive woody plant state shifts, determining the location and the nature of the limit between open and tree or shrub‐dominated landscapes. In order to adapt management to prevailing and future climate, we need to understand how browsing and climatic factors together affect the growth of plants at biome borders. To disentangle herbivory and climate effects, we combined long‐term tree growth monitoring and dendroecology to investigate woody plant growth under different temperatures and red deer (Cervus elaphus) herbivory pressures at forest–moorland ecotones in the Scottish highlands. Reforestation and deer densities are core and conflicting management concerns in the area, and there is an urgent need for additional knowledge. We found that deer herbivory and climate had significant and interactive effects on tree growth: in the presence of red deer, pine (Pinus sylvestris) growth responded more strongly to annual temperature than in the absence of deer, possibly reflecting differing plant–plant competition and facilitation conditions. As expected, pine growth was negatively related to deer density and positively to temperature. However, at the tree population level, warming decreased growth when more than 60% of shoots were browsed. Heather (Calluna vulgaris) growth was negatively related to temperature and the direction of the response to deer switched from negative to positive when mean annual temperatures fell below 6.0°C. In addition, our models allow estimates to be made of how woody plant growth responds under specific combinations of temperature and herbivory, and show how deer management can be adapted to predicted climatic changes in order to more effectively achieve reforestation goals. Our results support the hypothesis that temperature and herbivory have interactive effects on woody plant growth, and thus accounting for just one of these two factors is insufficient for understanding plant growth mechanics at biome transitions. Furthermore, we show that climate‐driven woody plant growth increases can be negated by herbivory. © 2020 The Authors. Ecological Applications published by Wiley Periodicals LLC on behalf of Ecological Society of America This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

International audience; Plants optimize their growth and survival through highly integrated regulatory networks that coordinate defensive measures and developmental transitions in response to environmental cues. Protein phosphatase 2A (PP2A) is a key signaling component that controls stress reactions and growth at different stages of plant development, and the PP2A regulatory subunit PP2A-B'gamma is required for negative regulation of pathogenesis responses and for maintenance of cell homeostasis in short-day conditions. Here, we report molecular mechanisms by which PP2A-B'gamma regulates Botrytis cinerea resistance and leaf senescence in Arabidopsis (Arabidopsis thaliana). We extend the molecular functionality of PP2A-B'gamma to a protein kinase-phosphatase interaction with the defense-associated calcium-dependent protein kinase CPK1 and present indications this interaction may function to control CPK1 activity. In presenescent leaf tissues, PP2A-B'gamma is also required to negatively control the expression of salicylic acid-related defense genes, which have recently proven vital in plant resistance to necrotrophic fungal pathogens. In addition, we find the premature leaf yellowing of pp2a-b'gamma depends on salicylic acid biosynthesis via SALICYLIC ACID INDUCTION DEFICIENT2 and bears the hallmarks of developmental leaf senescence. We propose PP2A-B'gamma age-dependently controls salicylic acid-related signaling in plant immunity and developmental leaf senescence.

Duchenne muscular dystrophy (DMD) is a fatal muscle‐wasting disease arising from mutations in the dystrophin gene. Upregulation of utrophin to compensate for the missing dystrophin offers a potential therapy independent of patient genotype. The first‐in‐class utrophin modulator ezutromid/SMT C1100 was developed from a phenotypic screen through to a Phase 2 clinical trial. Promising efficacy and evidence of target engagement was observed in DMD patients after 24 weeks of treatment, however trial endpoints were not met after 48 weeks. The objective of this study was to understand the mechanism of action of ezutromid which could explain the lack of sustained efficacy and help development of new generations of utrophin modulators. Using chemical proteomics and phenotypic profiling we show that the aryl hydrocarbon receptor (AhR) is a target of ezutromid. Several lines of evidence demonstrate that ezutromid binds AhR with an apparent KD of 50 nm and behaves as an AhR antagonist. Furthermore, other reported AhR antagonists also upregulate utrophin, showing that this pathway, which is currently being explored in other clinical applications including oncology and rheumatoid arthritis, could also be exploited in future DMD therapies., Chemical proteomics and phenotypic profiling illustrate the aryl hydrocarbon receptor (AhR) is a target of ezutromid, the first‐in‐class utrophin modulator for the treatment of Duchenne muscular dystrophy. Ezutromid binds to AhR with an apparent K D of 50 nm and functions as an AhR antagonist. Other reported AhR antagonists also upregulate utrophin, showing that this pathway, which is currently being explored in other clinical applications, could also be exploited in future DMD therapies.

Genome-wide association studies have identified breast cancer risk variants in over 150 genomic regions, but the mechanisms underlying risk remain largely unknown. These regions were explored by combining association analysis with in silico genomic feature annotations. We defined 205 independent risk-associated signals with the set of credible causal variants in each one. In parallel, we used a Bayesian approach (PAINTOR) that combines genetic association, linkage disequilibrium and enriched genomic features to determine variants with high posterior probabilities of being causal. Potentially causal variants were significantly over-represented in active gene regulatory regions and transcription factor binding sites. We applied our INQUSIT pipeline for prioritizing genes as targets of those potentially causal variants, using gene expression (expression quantitative trait loci), chromatin interaction and functional annotations. Known cancer drivers, transcription factors and genes in the developmental, apoptosis, immune system and DNA integrity checkpoint gene ontology pathways were over-represented among the highest-confidence target genes., This work was supported by the European Union’s Horizon 2020 Research and Innovation Programme under Marie Sklodowska-Curie grant agreement number 656144. Genotyping of the OncoArray was principally funded from three sources: the PERSPECTIVE project (funded by the Government of Canada through Genome Canada and the Canadian Institutes of Health Research, the ‘Ministère de l’Économie de la Science et de l’Innovation du Québec’ (through Genome Québec) and the Quebec Breast Cancer Foundation); the NCI Genetic Associations and Mechanisms in Oncology (GAME-ON) initiative and the Discovery, Biology and Risk of Inherited Variants in Breast Cancer (DRIVE) project (NIH grants U19 CA148065 and X01HG007492); and Cancer Research UK (C1287/A10118, C8197/A16565 and C1287/A16563). BCAC is funded by Cancer Research UK (C1287/A16563), by the European Community’s Seventh Framework Programme under grant agreement 223175 (HEALTH-F2-2009-223175) (COGS) and by the European Union’s Horizon 2020 Research and Innovation Programme under grant agreements 633784 (B-CAST) and 634935 (BRIDGES). Genotyping of the iCOGS array was funded by the European Union (HEALTH-F2-2009-223175), Cancer Research UK (C1287/A10710), the Canadian Institutes of Health Research for the ‘CIHR Team in Familial Risks of Breast Cancer’ program, and the Ministry of Economic Development, Innovation and Export Trade of Quebec (grant PSR-SIIRI-701). Combining of the GWAS data was supported in part by NIH Cancer Post-Cancer GWAS initiative grant U19 CA 148065 (DRIVE; part of the GAME-ON initiative).

Various biotic and abiotic stresses are threatening forests. Modern remote sensing technologies provide powerful means for monitoring forest health, and provide a sustainable basis for forest management and protection. The objective of this study was to develop unmanned aerial vehicle (UAV) based spectral remote sensing technologies for tree health assessment, particularly, for detecting the European spruce bark beetle (Ips typographus L.) attacks. Our focus was to study the early detection of bark beetle attack, i.e. the “green attack” phase. This is a difficult remote sensing task as there does not exist distinct symptoms that can be observed by the human eye. A test site in a Norway spruce (Picea abies (L.) Karst.) dominated forest was established in Southern-Finland in summer 2019. It had an emergent bark beetle outbreak and it was also suffering from other stress factors, especially the root and butt rot (Heterobasidion annosum (Fr.) Bref. s. lato). Altogether seven multitemporal hyper- and multispectral UAV remote sensing datasets were captured from the area in August to October 2019. Firstly, we explored deterioration of tree health and development of spectral symptoms using a time series of UAV hyperspectral imagery. Secondly, we trained assessed a machine learning model for classification of spruce health into classes of “bark beetle green attack”, “root-rot”, and “healthy”. Finally, we demonstrated the use of the model in tree health mapping in a test area. Our preliminary results were promising and indicated that the green attack phase could be detected using the accurately calibrated spectral image data.

Scots pine (Pinus sylvestris L.) is an important source of biogenic volatile organic compounds (BVOCs) in the boreal zone. BVOC emission rate and profile affect air quality, climate forcing, plant stress tolerance, and thus the growing conditions of forests. BVOC emission profile of shoots and forest floor, as well as emission rates from forest floor, were studied in a latitudinal provenance experiment with 19-year-old Scots pine common garden in Central Finland. The provenances studied were Saaremaa (SAA, 58°22′), Korpilahti (KOR, 62°0′), Suomussalmi (SUO, 65°10′), and Muonio (MUO, 67°56′). A chemotype with high proportion of Δ-3-carene, terpinolene, sabinene, γ-terpinene, and α-terpinene was significantly more common for the southern SAA than the northern SUO and MUO provenances. A chemotype with high proportion of α-pinene, β-pinene, limonene, and myrcene was more common in the three northernmost provenances. The main compounds emitted by forest floor were α-pinene, Δ-3-carene, and camphene. Similarly to shoot emissions, forest floor emissions from SAA had highest proportion of Δ-3-carene. Average total VOC emission rate from forest floor was 50 μg·m−2·h−1 at the end of August. Total emission rates were 65% higher in KOR than in MUO. High emission rates were explained by the high amount of decomposing needle litter and low moss coverage.

The immunosuppressive microenvironment in glioblastoma (GBM) prevents an efficient antitumoral immune response and enables tumor formation and growth. Although an understanding of the nature of immunosuppression is still largely lacking, it is important for successful cancer treatment through immune system modulation. To gain insight into immunosuppression in GBM, we performed a computational analysis to model relative immune cell content and type of immune response in each GBM tumor sample from The Cancer Genome Atlas RNA-seq data set. We uncovered high variability in immune system–related responses and in the composition of the microenvironment across the cohort, suggesting immunologic diversity. Immune cell compositions were associated with typical alterations such as IDH mutation or inactivating NF1 mutation/deletion. Furthermore, our analysis identified three GBM subgroups presenting different adaptive immune responses: negative, humoral, and cellular-like. These subgroups were linked to transcriptional GBM subtypes and typical genetic alterations. All G-CIMP and IDH-mutated samples were in the negative group, which was also enriched by cases with focal amplification of CDK4 and MARCH9. IDH1-mutated samples showed lower expression and higher DNA methylation of MHC-I–type HLA genes. Overall, our analysis reveals heterogeneity in the immune microenvironment of GBM and identifies new markers for immunosuppression. Characterization of diverse immune responses will facilitate patient stratification and improve personalized immunotherapy in the future. Significance: This study utilizes a computational approach to characterize the immune environments in glioblastoma and shows that glioblastoma immune microenvironments can be classified into three major subgroups, which are linked to typical glioblastoma alterations such as IDH mutation, NF1 inactivation, and CDK4-MARCH9 locus amplification. Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/78/19/5574/F1.large.jpg. Cancer Res; 78(19); 5574–85. ©2018 AACR.

We tested if root colonisation by ectomycorrhizal fungi (EMF) could alter the susceptibility of Norway spruce (Picea abies) seedlings to root rot infection or necrotic foliar pathogens. Firstly, spruce seedlings were inoculated by various EMF and challenged with Heterobasidion isolates in triaxenix tubes. The ascomycete EMF Meliniomyces bicolor, that had showed strong antagonistic properties towards root rot causing Heterobasidion in vitro, protected spruce seedlings effectively against root rot. Secondly, spruce seedlings, inoculated with M. bicolor or the forest humus, were subjected to necrotrophic foliar pathogens in conventional forest nursery conditions on peat substrates. Botrytis cinerea infection after winter was mild and the level of needle damage was independent of substrate and EMF colonisation. Needle damage severity caused by Gremminiella abietina was high in seedlings grown in substrates with high nutrient availability as well as in seedlings with well-established EMF communities. These results show that albeit M. bicolor is able to protect spruce seedlings against Heterobasidion root rot in axenic cultures it fails to induce systemic protection against foliar pathogens. We also point out that unsterile inoculum sources, such as the forest humus, should not be considered for use in greenhouse conditions as they might predispose seedlings to unintended needle damages.

Several known breast cancer susceptibility genes with moderate-to-high risk alleles encode proteins involved in DNA damage response (DDR). As these explain less than half of the hereditary breast cancer cases, additional predisposing alleles are likely to be discovered. Many of the previous studies utilizing massive parallel sequencing have focused on the protein-truncating variants, and the role of rare missense mutations has remained poorly addressed. To identify novel susceptibility factors, we have systematically analyzed the data from our parallel sequencing of 796 DDR genes in 189 Northern Finnish hereditary breast cancer patients for rare missense variants, predicted as deleterious. Thirty-five variants were studied here for the disease association using Finnish breast cancer case (n = 492-2,035) and control (n = 277-1,539) cohorts. As a result, two missense variants in genes involved in DNA replication, RECQL p.I156M and POLG p.L392V, the former involving genomic and the latter mitochondrial DNA replication, showed significant association with risk of breast cancer. Rare RECQL p.I156M allele was observed in breast cancer cases only (6/1,946, 0.3%, p = 0.043), whereas POLG p.L392V was two times more frequent in breast cancer cases (53/2,238, 2.4%) compared to controls (18/1,539, 1.2%, OR = 2.1, 95% CI 1.2-3.5, p = 0.010). Based on the current genetic data, both RECQL p.I156M and POLG p.L392V represent novel breast cancer predisposing alleles.

Understanding the chemical structure of lignin in willow bark is an indispensable step to design how to separate its fiber bundles. The whole cell wall and enzyme lignin preparations sequentially isolated from ball-milled bark, inner bark, and wood were comparatively investigated by nuclear magnetic resonance (NMR) spectroscopy and three classical degradative methods, i.e., alkaline nitrobenzene oxidation, derivatization followed by reductive cleavage, and analytical thioacidolysis. All results demonstrated that the guaiacyl (G) units were predominant in the willow bark lignin over syringyl (S) and minor p-hydroxyphenyl (H) units. Moreover, the monomer yields and S/G ratio rose progressively from bark to inner bark and wood, indicating that lignin may be more condensed in bark than in other tissues. Additionally, major interunit linkage substructures (β-aryl ethers, phenylcoumarans, and resinols) together with cinnamyl alcohol end groups were relatively quantitated by two-dimensional NMR spectroscopy. Bark and inner bark were rich in pectins and proteins, which were present in large quantities and also in the enzyme lignin preparations.

In recent treatments of Fissidens subgenus Fissidens, the species delimitation of F. bryoides, the type species of the subgenus, has been rather variable. In Finland, F. bryoides has either been split into F. bryoides, F. gymnandrus and F. viridulus, or treated as a single species, F. bryoides s. lato, including two or more intraspecific taxa. The herbarium material of Finnish F. bryoides, F. gymnandrus and F. viridulus was studied to clarify the distribution and ecological requirements of the three taxa and to find out which morphological and ecological characters are best for their delimitation. The phylogeny lends support to recognizing F. bryoides s. lato including F. gymnandrus and F. viridulus as F. bryoides var. gymnandrus and F. bryoides var. viridulus. The phylogeny reconstruction based on 18 morphological characters that are commonly used to distinguish the taxa could not resolve F. bryoides, F. gymnandrus and F. viridulus as monophyletic species. However, F. bryoides s. lato including all herbarium material of the three species was resolved as a monophyletic group morphologically distinct from the closely related F. pusillus. The synapomorphic characters that distinguish F. bryoides s. lato are the percurrent costa, limbidia that reach the leaf acumen, the acuminate leaf acumen, and a preference for substrates other than rock. We found differences in habitat requirements between the varieties, while sexual systems sometimes vary even within a single specimen. Fissidens bryoides var. bryoides and F. bryoides var. gymnandrus appear to have wider distribution in Finland than expected. Unlike F. bryoides var. viridulus the number of localities of F. bryoides var. bryoides and F. bryoides var. gymnandrus is low. Information on their conservation status is also provided.

Aim To assess the role of human bocavirus 1 (HBoV1) as a causative agent of non‐severe community‐acquired pneumonia (CAP) in children. Methods Patients aged 2‐59 months with non‐severe CAP (respiratory complaints and radiographic pulmonary infiltrate/consolidation) attending a University Hospital in Salvador, Brazil were enrolled in a prospective cohort. From 820 recruited children in a clinical trial (http://ClinicalTrials.gov NCT01200706), nasopharyngeal aspirate (NPA), and acute and convalescent serum samples were obtained from 759 (92.6%) patients. NPAs were tested for 16 respiratory viruses by PCR. Acute HBoV1 infection was confirmed by measuring specific IgM and IgG responses in paired serum samples. Results Respiratory viruses were detected in 693 (91.3%; 95%CI: 89.1‐93.2) CAP cases by PCR. HBoV1‐DNA was detected in 159 (20.9%; 95%CI: 18.2‐24.0) cases. Of these 159 PCR positive cases, acute HBoV1 infection was confirmed serologically in 38 cases (23.9%; 95%CI: 17.8‐31.0). Overall, acute HBoV1 infection was confirmed in 5.0% (38/759) of non‐severe CAP patients. HBoV1 was detected in 151 cases with at least one other virus making 31.7% of all multiple virus (n = 477) detections. Among all 759 cases, 216 had one respiratory virus detected, and sole HBoV1 was detected in only 8 (3.7%). Acute HBoV1 infection was serologically diagnosed in 34 (22.5%) HBoV1‐DNA‐positive cases with another virus, compared to 4 (50.0%) cases with sole virus detection (p = 0.09). Conclusion HBoV1 was detected by PCR in one fifth of the children with non‐severe CAP and acute HBoV1 infection was serologically confirmed in one quarter of these cases.

Funding Information: The authors would like to thank Pitk?nen Leena, Hatakka Rita and Linnekoski Juha for their skilful assistance in HP-SEC, HPAEC-PAD and GC-FID measurements. This work made use of the Aalto University NMR premises. This work was a part of the Academy of Finland's Flagship Programme under Projects No. 318890 and 318891 (Competence Center for Materials Bioeconomy, FinnCERES). Publisher Copyright: © 2021 The Author(s) Aiming to understand the importance of debarking on the controlled utilization of phenolic-rich willow biomass, biobutanol was produced from it by using Clostridium acetobutylicum. Acid-catalysed steam explosion and enzymatic hydrolysis (EH) were investigated before the acetone-butanol-ethanol (ABE) fermentation. The hydrolysable sugar yield and ABE fermentation efficiency were found to decline progressively from willow wood (WW) to HWE WB (hot water extracted willow biomass), WB (willow biomass) and the WW + HWE (willow wood plus the artificial willow bark water extracts), indicating that the pre-removal of water extracts or the bark can significantly improve ABE yield. Notably, the ABE productivity of WW achieved 12.7 g/L at the solvent yield of 31%, and the butanol concentration (i.e. 8.5 g/L) generated by WW is relatively high among the reported lignocellulosic-derived biomass. Additionally, it is hypothesized that under acidic conditions and high temperatures the fructose present in willow water extractsform hydroxymethylfurfural during steam explosion, which then spontaneously condenses with phenolic substances of willow bark to form a solid furanic precipitate. The formed furanic precipitates play inhibitory role in the enzymatic hydrolysis and are thereby deleterious to the ABE fermentation.

A literature survey identified 30 articles providing data on the concentrations of organohalogen compounds in fish caught from the Gulf of Finland: polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs), the chlorinated pesticide DDT and polybrominated diphenylethers (PBDEs). Results were found for 14 fish species in total, but mainly for herring (Clupea harengus), salmon (Salmo salar) and sprat (Sprattus sprattus), and the oldest data were for PCBs, from 1975. PCBs and PCDDs were the most prevalent organohalogens in the Gulf of Finland compared to other areas of the Baltic Sea. In particular, more local fish species and benthic feeders caught from the eastern parts of the Gulf of Finland appear to contain higher concentrations of organochlorines than those from the western parts. The concentration of dioxins plus dioxin-like PCBs as WHO-TEQPCDD/F + PCB in herring has demonstrated a decreasing trend since the late 1970s. However, the maximum allowable concentration of WHO-TEQPCDD/F + PCB in fish set by the EC was exceeded in all salmon and older herring specimens and in some whitefish (Coregonus lavaretus), bream (Abramis brama), sea trout (Salmo trutta) and flounder (Platichthys flesus). The respective limit concentration for fish used as animal feed was exceeded in most of the fish samples.

Impaired 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2)-dependent cortisol inactivation can lead to electrolyte dysbalance, hypertension and cardiometabolic disease. Furthermore, placental 11β-HSD2 essentially protects the fetus from high maternal glucocorticoid levels, and its impaired function has been associated with altered fetal growth and a higher risk for cardio-metabolic diseases in later life. Despite its important role, 11β-HSD2 is not included in current off-target screening approaches. To identify potential 11β-HSD inhibitors amongst approved drugs, a pharmacophore model was used for virtual screening, followed by biological assessment of selected hits. This led to the identification of several azole fungicides as 11β-HSD inhibitors, showing a significant structure-activity relationship between azole scaffold size, 11β-HSD enzyme selectivity and inhibitory potency. A hydrophobic linker connecting the azole ring to the other, more polar end of the molecule was observed to be favorable for 11β-HSD2 inhibition and selectivity over 11β-HSD1. The most potent 11β-HSD2 inhibition, using cell lysates expressing recombinant human 11β-HSD2, was obtained for itraconazole (IC(50) 139 ± 14 nM), its active metabolite hydroxyitraconazole (IC(50) 223 ± 31 nM) and posaconazole (IC(50) 460 ± 98 nM). Interestingly, experiments with mouse and rat kidney homogenates showed considerably lower inhibitory activity of these compounds toward 11β-HSD2, indicating important species-specific differences. Thus, 11β-HSD2 inhibition by these compounds are likely to be overlooked in preclinical rodent studies. Inhibition of placental 11β-HSD2 by these compounds, in addition to the known inhibition of cytochrome P450 enzymes and P-glycoprotein efflux transport, might contribute to elevated local cortisol levels, thereby affecting fetal programming.

Summary Objectives Besides group A streptococcus (GAS), microbial causes of pharyngitis in children are not well known. We aimed to document the viral and bacterial aetiology of pharyngitis and to assess the pathogenic role of viruses by determining the myxovirus resistance protein A (MxA) in the blood as a marker of interferon response. Methods In this prospective observational study, throat swabs and blood samples were collected from children (age 1–16 years) presenting to the emergency department with febrile pharyngitis. Microbial cause was sought by bacterial culture, polymerase chain reaction, and serology. Blood MxA level was determined. Results A potential pathogen was detected in 88% of 83 patients: GAS alone in 10%, GAS and viruses in 13%, group C or G streptococci alone in 2% and together with viruses in 3%, and viruses alone in 59% of cases. Enteroviruses, rhinoviruses, and adenoviruses were the most frequently detected viruses. Blood MxA levels were higher in children with viral (880 [245–1250] μg/L; median [IQR]) or concomitant GAS-viral (340 [150–710] μg/L) than in those with sole GAS (105 [80–160] μg/L) infections. Conclusions Detection of respiratory viruses simultaneously with elevated blood MxA levels supports the causative role of viruses in the majority of children with pharyngitis., Highlights • We evaluated the microbiological aetiology of febrile pharyngitis in 83 children. • A potential pathogen could be detected in 88% and virus in 76% of patients. • Blood myxovirus resistance protein A (MxA) levels were elevated in most of the patients with virus finding. • MxA is a promising biomarker of virus infection.

Fatty acids in muscle tissue and eggs of female Atlantic salmon Salmo salar spawners were analysed to evaluate the dietary quality of their final feeding areas in the Baltic Sea. The final likely feeding area was identified by comparing stable carbon and nitrogen isotope composition of the outermost growth region (final annulus) of scales of returned S. salar with that of reference S. salar caught from different feeding areas. Some overlap of stable-isotope reference values among the three areas, in addition to prespawning fasting, decreased the ability of muscle tri-acylglycerols to discriminate the final likely feeding area and the area's dietary quality. Among three long-chained polyunsaturated fatty acids, docosahexaenoic acid (DHA; 22:6n-3), eicosapentaenoic acid (EPA; 20:5n-3) and arachidonic acid (ARA; 20:4n-6), the proportions of ARA in total lipids of spawning S. salar muscle and eggs showed a significant negative correlation with increasing probability of S. salar having returned from the Baltic Sea main basin (i.e. the Baltic Sea proper). The results suggest that ARA in muscle and eggs is the best dietary indicator for dietary characteristics of final marine feeding area dietary characteristics among S. salar in the Baltic Sea.

Karyopherin alpha 7 (KPNA7) belongs to a family of nuclear import proteins that recognize and bind nuclear localization signals (NLSs) in proteins to be transported to the nucleus. Previously we found that KPNA7 is overexpressed in a subset of pancreatic cancer cell lines and acts as a critical regulator of growth in these cells. This characteristic of KPNA7 is likely to be mediated by its cargo proteins that are still mainly unknown. Here, we used protein affinity chromatography in Hs700T and MIA PaCa-2 pancreatic cancer cell lines and identified 377 putative KPNA7 cargo proteins, most of which were known or predicted to localize to the nucleus. The interaction was confirmed for two of the candidates, MVP and ZNF414, using co-immunoprecipitation, and their transport to the nucleus was hindered by siRNA based KPNA7 silencing. Most importantly, silencing of MVP and ZNF414 resulted in marked reduction in Hs700T cell growth. In conclusion, these data uncover two previously unknown human KPNA7 cargo proteins with distinct roles as novel regulators of pancreatic cancer cell growth, thus deepening our understanding on the contribution of nuclear transport in cancer pathogenesis.

Background The prognosis of glioblastoma remains poor, related to its diffuse spread within the brain. There is an ongoing search for molecular regulators of this particularly invasive behavior. One approach is to look for actin regulating proteins that might be targeted by future anti-cancer therapy. The formin family of proteins orchestrates rearrangement of the actin cytoskeleton in multiple cellular processes. Recently, the formin proteins mDia1 and mDia2 were shown to be expressed in glioblastoma in vitro, and their function could be modified by small molecule agonists. This finding implies that the formins could be future therapeutic targets in glioblastoma. Methods In cell studies, we investigated the changes in expression of the 15 human formins in primary glioblastoma cells and commercially available glioblastoma cell lines during differentiation from spheroids to migrating cells using transcriptomic analysis and qRT-PCR. siRNA mediated knockdown of selected formins was performed to investigate whether their expression affects glioblastoma migration. Using immunohistochemistry, we studied the expression of two formins, FHOD1 and INF2, in tissue samples from 93 IDH-wildtype glioblastomas. Associated clinicopathological parameters and follow-up data were utilized to test whether formin expression correlates with survival or has prognostic value. Results We found that multiple formins were upregulated during migration. Knockdown of individual formins mDia1, mDia2, FHOD1 and INF2 significantly reduced migration in most studied cell lines. Among the studied formins, knockdown of INF2 generated the greatest reduction in motility in vitro. Using immunohistochemistry, we demonstrated expression of formin proteins FHOD1 and INF2 in glioblastoma tissues. Importantly, we found that moderate/high expression of INF2 was associated with significantly impaired prognosis. Conclusions Formins FHOD1 and INF2 participate in glioblastoma cell migration. Moderate/high expression of INF2 in glioblastoma tissue is associated with worse outcome. Taken together, our in vitro and tissue studies suggest a pivotal role for INF2 in glioblastoma. When specific inhibiting compounds become available, INF2 could be a target in the search for novel glioblastoma therapies.

Firefly luciferase (FLuc) is a powerful tool for molecular and cellular biology, and popular in high-throughput screening and drug discovery. However, FLuc assays have been plagued with positive and negative artefacts due to stabilisation and inhibition by small molecules from a range of chemical classes. Here we disclose Phase II clinical compound SMT C1100 for the treatment of Duchenne muscular dystrophy as an FLuc inhibitor (KD of 0.40 ± 0.15 µM). Enzyme kinetic studies using SMT C1100 and other non-competitive inhibitors including resveratrol and NFκBAI4 identified previously undescribed modes of inhibition with respect to FLuc’s luciferyl adenylate intermediate. Employing a photoaffinity strategy to identify SMT C1100′s binding site, a photolabelled SMT C1100 probe instead underwent FLuc-dependent photooxidation. Our findings support novel binding sites on FLuc for non-competitive inhibitors.

In this study, we present willow wood as a new low-cost, renewable, and sustainable biomass source for the production of a highly porous activated carbon for application in energy storage devices. The obtained activated carbon showed favorable features required for excellent electrochemical performance such as high surface area (∼2 800 m2 g-1) and pore volume (1.45 cm3 g-1), with coexistence of micropores and mesopores. This carbon material was tested as an electrode for supercapacitor application and showed a high specific capacitance of 394 F g-1 at a current density of 1 A g-1 and good cycling stability, retaining ∼94% capacitance after 5000 cycles (at a current density of 5 A g-1) in 6 M KOH electrolyte. The prepared carbon material also showed an excellent rate performance in a symmetrical two-electrode full cell configuration using 1 M Na2SO4 electrolyte, in a high working voltage of 1.8 V. The maximum energy density and power density of the fabricated symmetric cell reach 23 W h kg-1 and 10 000 W kg-1,respectively. These results demonstrate that willow wood can serve as a low-cost carbon feedstock for production of high-performance electrode material for supercapacitors.

Background In children suffering from severe lower airway illnesses, respiratory virus detection has given good prognostic information, but such reports in the elderly are scarce. Therefore, our aim was to study whether the detection of nasopharyngeal viral pathogens and conventional inflammatory markers in the frail elderly correlate to the presence, signs and symptoms or prognosis of radiographically-verified pneumonia. Methods Consecutive episodes of hospital care of patients 65 years and older with respiratory symptoms (N = 382) were prospectively studied as a cohort. Standard clinical questionnaire was filled by the study physician. Laboratory analyses included PCR diagnostics of nasopharyngeal swab samples for 14 respiratory viruses, C-reactive protein (CRP) and white blood cell count (WBC). Chest radiographs were systematically analysed by a study radiologist. The length of hospital stay, hospital revisit and death at ward were used as clinical endpoints. Results Median age of the patients was 83 years (range 76–90). Pneumonia was diagnosed in 112/382 (29%) of the studied episodes. One or more respiratory viruses were detected in 141/382 (37%) episodes and in 34/112 (30%) episodes also diagnosed with pneumonia. Pneumonia was associated with a WBC over 15 × 109/L (P = .006) and a CRP value over 80 mg/l (P

Red band needle blight of pines caused by Mycosphaerella pini (anamorph Dothistroma septosporum) has recently been recorded on Scots pine (Pinus sylvestris) at 14 rural districts in southern and central Finland. Scots pine is the most common and commercially most important tree species in Finland. Red bands with aggregations of conidial stromata on otherwise brown attached needles were frequently encountered on saplings and young trees in dense stands and sporadically on lower twigs of mature trees within 2 m of the ground. These symptoms and signs, typical for M. pini (1), were also observed on needles of P. contorta and P. cembra, which occur in Finland in low frequency. Symptoms of red band needle blight and abundant conidial stromata were found in March and April of 2008 after a mild and rainy winter. After this time, the frequency at which fresh acervuli were observed decreased. Conidia were isolated after squeezing conidial stromata into a drop of sterile water and rinsing out the drop onto water agar from where single conidia were picked up from under the microscope with a modified Pasteur pipette. The conidia were hyaline, smooth, thin walled, filiform, 2.0 to 2.7 (2.4) μm wide, and 15 to 37 (29.4) μm long. Germination of the conidia on water agar was 100%. The cultures grew slowly and reached a diameter of 4 to 10 mm within 3 weeks on modified orange serum agar (2) at 20°C and abundantly produced conidia. Complete sequences of the internal transcribed spacer (ITS) region including the 5.8S rRNA gene were obtained for three isolates from different rural districts: Hartola (61°34′N, 26°01′E), Kangasniemi (61°59′N, 26°39′E), and Suonenjoki (62°37′N, 27°07′E). These sequences are deposited in GenBank with Accession Nos. EU834294, EU834295, and EU834296 and are identical to each other and to more than 50 M. pini sequences in GenBank, including those of several Estonian and Austrian isolates. These isolates are deposited in the culture collection of the Finnish Forest Research Institute with identifiers Dot1, Dot2, and Dot8. Symptoms were reproduced after inoculation of 1-year-old Scots pine seedlings growing in seedling trays with 115 cm3 cavities. A conidial suspension (5 to 6·× 103 conidia ml–1) was prepared from two single-conidial cultures (Dot10 and Dot15), combined, and sprayed during July 2008 onto 192 seedlings until needles looked completely wet. Sixty-four seedlings were sprayed with distilled water as a control treatment. Seedlings were incubated outdoors in Suonenjoki and covered with a transparent plastic hood for the first 5 days after inoculation. The first symptoms (brown segments and red bands on needles) appeared on inoculated seedlings 1 month later, and conidial stromata appeared after another 2 to 4 weeks. M. pini was reisolated from the acervuli of 10 sample needles. Three months after inoculation, all inoculated seedlings showed symptoms while all noninoculated seedlings were healthy. It is possible that M. pini has spread recently from the south since it was identified for the first time in Estonia 2 years earlier (3). Although the Gulf of Finland separates Finnish pine forests from Estonian pine forests by approximately 50 to 100 km, spores may have been aerially disseminated over this distance. Alternatively, introduction of M. pini to Finland may have occurred on imported seedlings. References: (1) Anonymous. OEPP/EPPO Bull. 35, 303, 2005. (2) M. Hanso and R. Drenkhan. Plant Pathol. 57:170, 2008. (3) M. M. Müller et al. Mycol. Res. 98:593, 1994.

During a survey conducted in August 2009 in northern Norway, symptoms typical for red band needle blight (1) were observed in four young Scots pine (Pinus sylvestris) stands. The stands, less than 15 years old, were located in humid sites near rivers in Bardu and Målselv municipalities, Troms County. Many of the oldest needles (2- to 3-yearsold) in the lower part of young trees were partially or completely brown, but still attached, and red bands could be observed. Aggregations of conidial stromata were often seen in the red bands. Conidia were hyaline, smooth, thin walled, and filiform, 1.9 to 2.6 μm wide and 12 to 36 μm long. Isolations were made from conidiomata and the identity of Mycosphaerella pini was confirmed by partial sequencing of the internal transcribed spacer (ITS) region of a sample from Målselv (69°00′N, 18°51′E) (GenBank Accession No. JF796109). In June 2010, a survey was done in southeastern Norway where nearly 100 stands with young Scots pine, up to 20 years old, were inspected and typical symptoms of red band needle blight were observed in 10 stands in three municipalities in Hedmark County (Eidskog, Kongsvinger, and Trysil), less than 25 km from the Swedish border, and in one stand in Buskerud County (Nedre Eiker). Lower branches of up to 5 m tall trees had symptoms of red band needle blight, mainly on the oldest needles. In the red bands, acervuli of Dothistroma septosporum, the anamorph of M. pini, appeared. Typical conidia from acervuli were germinated in water agar and resulting mycelium was transferred to modified orange serum agar. Two isolates were sequenced as above and both herbarium samples and isolates (Kongsvinger, 60°06′N, 12°04′E; GenBank Accession No. JF796108 and Trysil, 61°14′N, 12°22′E; GenBank Accession No. JF796107) were deposited at the Norwegian Forest and Landscape Institute. To fulfill Koch's postulates, symptoms were reproduced by artificial inoculation onto 1-year-old Scots pine seedlings. In June 2010, a pooled conidial suspension (2.4 to 3.8 × 103 conidia per ml) from two single-conidium cultures was sprayed to runoff onto 128 seedlings and 64 seedlings were mock sprayed with distilled water. First symptoms (brown segments and red bands) appeared on inoculated seedlings 1 month later and acervuli appeared after another 6 to 8 weeks. M. pini was reisolated from the acervuli. Three months after inoculation, 90% of inoculated seedlings showed symptoms while all uninoculated seedlings were healthy. Damage caused by M. pini has increased in the northern hemisphere during the last 15 to 20 years, possibly because of climate change (4). In 2006, the fungus was recorded in Estonia (2), in 2008 in Finland (3), and by 2009, the disease had spread over large areas in Finland including the region near the Norwegian border in the north. The disease has been in Sweden for a few years (J. Stenlid, personnel communication). In southern Norway, the disease has mainly been observed near the Swedish border so here the disease may originate from Sweden. So far, however, only small areas of Norway have been surveyed. Both natural spread and human transport dissemination may be occurring. References: (1) Anonymous. OEPP/EPPO Bull. 35, 303, 2005. (2) M. Hanso and R. Drenkhan. Plant Pathol. 57:170, 2008. (3) M. Müller et al. Plant Dis. 93:322, 2009. (4) A. Woods et al. BioScience 55:761, 2005.

Genome-wide association studies have identified breast cancer risk variants in over 150 genomic regions, but the mechanisms underlying risk remain largely unknown. These regions were explored by combining association analysis with in silico genomic feature annotations. We defined 205 independent risk-associated signals with the set of credible causal variants (CCVs) in each one. In parallel, we used a Bayesian approach (PAINTOR) that combines genetic association, linkage disequilibrium, and enriched genomic features to determine variants with high posterior probabilities (HPPs) of being causal. Potentially causal variants were significantly over-represented in active gene regulatory regions and transcription factor binding sites. We applied our INQUSIT pipeline for prioritizing genes as targets of potentially causal variants, using gene expression (eQTL), chromatin interaction and functional annotations. Known cancer drivers, transcription factors and genes in the developmental, apoptosis, immune system and DNA integrity checkpoint gene ontology pathways, were over-represented among the 178 highest confidence target genes.