Epitranscriptomic modifications in RNA can dramatically alter the way our genetic code is deciphered. Cells utilize these modifications not only to maintain physiological processes, but also to respond to extracellular cues and various stressors. Most often, adenosine residues in RNA are targeted, and result in modifications including methylation and deamination. Such modified residues as N-6-methyl-adenosine (m6A) and inosine, respectively, have been associated with cardiovascular diseases, and contribute to disease pathologies. The Ischemic Heart Disease Epitranscriptomics and Biomarkers (IHD-EPITRAN) study aims to provide a more comprehensive understanding to their nature and role in cardiovascular pathology. The study hypothesis is that pathological features of IHD are mirrored in the blood epitranscriptome. The IHD-EPITRAN study focuses on m6A and A-to-I modifications of RNA. Patients are recruited from four cohorts: (I) patients with IHD and myocardial infarction undergoing urgent revascularization; (II) patients with stable IHD undergoing coronary artery bypass grafting; (III) controls without coronary obstructions undergoing valve replacement due to aortic stenosis and (IV) controls with healthy coronaries verified by computed tomography. The abundance and distribution of m6A and A-to-I modifications in blood RNA are charted by quantitative and qualitative methods. Selected other modified nucleosides as well as IHD candidate protein and metabolic biomarkers are measured for reference. The results of the IHD-EPITRAN study can be expected to enable identification of epitranscriptomic IHD biomarker candidates and potential drug targets.

Chemical modifications of RNA provide an additional, epitranscriptomic, level of control over cellular functions. N-6-methylated adenosines (m6As) are found in several types of RNA, and their amounts are regulated by methyltransferases and demethylases. One of the most important enzymes catalyzing generation of m6A on mRNA is the trimer N-6-methyltransferase METTL3-14-WTAP complex. Its activity has been linked to such critical biological processes as cell differentiation, proliferation, and death. We used in silico-based discovery to identify small-molecule ligands that bind to METTL3-14-WTAP and determined experimentally their binding affinity and kinetics, as well as their effect on enzymatic function. We show that these ligands serve as activators of the METTL3-14-WTAP complex.

Mikrobilääkkeille resistenttien bakteerikantojen yleistyminen on kasvava maailmanlaajuinen ongelma. Suomessa antibioottien käyttö tuotantoeläimillä on useisiin muihin maihin nähden vähäistä, mutta silti potentiaalinen riski. Lääkittyjen eläinten lantaan erittyy mikrobilääkejäämiä ja resistenttejä bakteerikantoja, ja lannan lannoitekäyttö voikin olla riski niiden leviämiseen peltoon ja edelleen luontoympäristöön. Lannan tehokas käyttö edistää ravinnekiertoa maataloudessa ja Suomikin tavoittelee lisää lannan prosessointia. Keskitetyistä laitoksista lääkejäämät ja resistentit bakteerikannat voivat levitä entistä laajemmalle. Toisaalta prosessointi voi mahdollistaa lääkejäämien ja resistenttien kantojen tuhoutumisen. Biokaasuprosessi mahdollistaa yhtäaikaisen lannan ravinteiden ja energian hyödyntämisen. Mikrobilääkkeet voivat kuitenkin häiritä sen mikrobiologiaa. Toisaalta lääkeaineet voivat myös hajota täysin tai osin prosessin aikana. Kahden Suomessa yleisesti eläinlääkityksessä käytetyn antibiootin, oksitetrasykliinin ja siprofloksasiinin, vaikutusta lypsykarjan lietelannan metaanintuottoon tutkittiin eri pitoisuuksina ja seoksina. Kokeella sel-vitettiin, miten ja millä pitoisuuksilla antibiootit vaikuttavat metaanintuottoa alentavasti. Tutkimus toteutettiin 30 vrk panostoimisella kokeella käyttäen automatisoitua metaanipotentiaalin mit-tauslaitteistoa AMPTS II (Bioprocess Control AB, Ruotsi). Kokeessa seurattiin lypsykarjan lietelannan hajoamista ja ymppinä oli maatilan biokaasulaitoksenmädäte. Antibiootit lisättiin lannan ja ympin seokseen vesiliuoksina. Kokeessa oli 14 koejäsentä. Lietelantakontrollin ja pelkän ympin lisäksi oli 12 erilaisia antibioottikäsittelyjä: 4 oksitetrasykliinipitoisuutta (10–500 mg kg-1), 4 siprofloksasiinipitoisuutta (2–50 mg kg-1) sekä 4 seosta, joissa siprofloksasiinipitoisuus oli 10 ja 20 mg kg-1 ja oksitetrasykliinipitoisuus 50 ja 100 mg kg-1. Lietelannan metaanipotentiaali oli 218 m3CH4 MgVS-1 (kontrolli). Eniten lannan hajoamista esti oksitetra-sykliinipitoisuus 500 mg kg-1, joka tuotti metaania 48% vähemmän, 105 m3CH4 MgVS-1. Oksitetrasykliinin pitoisuudella 10 mg kg-1 ei havaittu vaikutusta metaanintuottoon, mutta seuraavaksi suuremmalla annoksella 50 mg kg-1 metaanintuotto heikkeni selvästi (170 m3CH4 MgVS-1). Samoin siprofloksasiinin pienimmällä annoksella 2 mg kg-1 ei vaikutusta havaittu, mutta 10 mg kg-1 vähensi metaanintuottoa (197 m3CH4 MgVS-1). Seoskäsittelyiden pienimmällä tutkitulla pitoisuusyhdistelmällä (oksitetrasykliini 50 mg kg-1 + siprofloksasiini 10 mg kg-1) metaanintuotto oli 161 m3CH4 MgVS-1. Antibioottien vaikutus metaanintuottoon oli selkeä tiettyjen pitoisuuksien ylittyessä; mitä suurempi antibioottiannostus oli, sitä vähemmän metaania muodostui. Tutkitut antibioottipitoisuudet olivat kuitenkin selvästi korkeampia kuin suomalaisessa lypsykarjan lietelannassa samassa tutkimuksessa havaittiin (oksi-tetrasykliini 5 μg kg-1, siprofloksasiini alle määritysrajan 3 μg kg-1). Näillä pitoisuuksilla metaanintuoton ei havaittu häiriintyvän.

The most common rodent control method worldwide is anticoagulant rodenticides (ARs), which cause death by internal bleeding. ARs can transfer to non-target predators via secondary exposure, i.e. by consuming contaminated rodents. Here we quantify the prevalence of seven AR substances in the liver tissues of altogether 17 mammalian or avian predator or scavenger species in Finland. In addition, we identify the environmental and biological factors potentially linked to secondary AR poisoning. No previous AR screenings have been conducted in the country, despite the widespread use of ARs and their potential impacts on the high levels of the ecosystem food chain. ARs were detected (≥0.3 μg/kg) in 82% of the 131 samples. The most prevalent and the AR with highest concentrations was bromadiolone (65% of samples). In 77% of the positive samples more than one (2-5) different ARs were detected. Of the environmental variables, we only found a weakly positive relationship between the coumatetralyl concentration and the livestock farm density. Conversely, overall AR concentration and number, as well as the concentration of three separate ARs (coumatetralyl, difenacoum and bromadiolone) differed among the three species groups tested, with the group "other mammals" (largely represented by red fox and raccoon dog) having higher values than the groups presented by mustelids or by birds. ARs are authorized only as biocides in Finland and a national strategy on risk management (e.g. for minimising secondary poisoning of non-target species) of ARs was adopted in 2011. Based on these results it appears that the risk mitigation measures (RMMs) either have not been followed or have not been effective in preventing wide scale secondary exposure. Continued monitoring of AR residues in non-target species is needed in order to evaluate the effectiveness of current RMMs and a need for new ones to reduce the risk of secondary poisoning.

Female perch (Perca fluviatilis) were sampled annually in late summer from 2006 to 2009 from the open sea of the eastern Gulf of Finland off Haapasaari island to monitor baseline biliary PAH metabolite concentrations. In addition, two coastal locations were sampled in 2008. PAH metabolite concentrations were compared between the open sea and coastal samples and between the sampling years and examined in relation to the body characteristics of perch. Of the PAH metabolites, only 1-hydroxypyrene (1-OH pyrene) was detected at quantifiable levels in the bile of nearly all perch individuals. There were some annual differences but no temporal trend in the concentration of biliary 1-OH pyrene in perch from Haapasaari. At the coastal locations, 1-OH pyrene concentrations in the bile of perch were significantly higher than in the open sea Haapasaari area, probably due to greater contamination of the coastal sites and differences in feeding behaviour. No correlations between the body characteristics of perch and 1-OH pyrene concentrations were detected. It is concluded that PAH metabolites in the bile of fish could be measured in the Gulf of Finland to detect oil spills in the open sea, and the cost-effective total fluorescence method could be used in such monitoring programmes.

The mycotoxin enniatin B, a cyclic hexadepsipeptide produced by the plant pathogen Fusarium, is prevalent in grains and grain-based products in different geographical areas. Although enniatins have not been associated with toxic outbreaks, they have caused toxicity in vitro in several cell lines. In this study, the cytotoxic effects of enniatin B were assessed in relation to cellular energy metabolism, cell proliferation, and the induction of apoptosis in Balb 3T3 and HepG2 cells. The mechanism of toxicity was examined by means of whole genome expression profiling of exposed rat primary hepatocytes. Enniatin B altered cellular energy metabolism and reduced cell proliferation in Balb 3T3 and HepG2 cell lines. Furthermore, the proportion of apoptotic cell populations of Balb 3T3 cells slightly increased. On the other hand, enniatin B caused necrotic cell death in primary hepatocytes. Gene expression studies revealed the alteration of energy metabolism due to effects on mitochondrial organization and function and the assembly of complex I of the electron transport chain.

Moniliformin is a Fusarium mycotoxin mainly produced by several species infecting grains in different climatic conditions. According to our previous studies, it is acutely toxic to rats, with an LD50 cut-off value of 25 mg/kg b.w. To further assess the possible health risks of low dose exposure to moniliformin, a subacute oral toxicity study was conducted in Sprague-Dawley rats, adapting OECD guideline 407. Five dose groups and two satellite groups, each consisting of five male rats, were daily exposed to moniliformin by gavage. Two rats in the highest dose group, showed decreased activity followed by acute heart failure and death. The rats of the lower doses (<9 mg/kg b.w.) showed no signs of toxicity. The daily intake of moniliformin strongly reduced the phagocytic activity of neutrophils in all dose groups. The decrease continued in the satellite group during the follow-up period, indicating a severe impact on the immune system and a LOAEL value of 3 mg/kg b.w. for moniliformin. Moniliformin was rapidly excreted into urine, ranging between 20.2 and 31.5% daily and showed no signs of accumulation. The concentration of moniliformin in faeces was less than 2%, which suggests efficient absorption from the gastrointestinal tract.

Copine VI (CPNE6) is a member of copines family, a calcium-dependent phospholipids-binding protein group found in many diverse eukaryotic organisms. Although earlier studies have shown that CPNE6 is almost exclusively expressed in brain, the exact biological functions remain unclear. The purpose of this study is to explore the relationship between epilepsy and CPNE6 expression. In present study, we investigated the expression pattern and distribution of CPNE6 in patients with refractory epilepsy and in a chronic pilocarpine-induced epileptic rat model by quantitative real-time PCR, Western blot and immunofluorescence. The results showed that the expression of CPNE6 increased remarkably in epileptic patients and in experimental epileptic rats. Double immunofluorescence labeling studies have revealed that CPNE6 protein is mainly expressed in neurons, demonstrated by co-localization with the dendritic marker, MAP2. Our results are the first to indicate that the abnormal expression of the CPNE6 in epileptic brain tissue may play an important role in epilepsy, especially refractory epilepsy.

•The subacute oral toxicity of synthetic moniliformin in rats was assessed according to an adaption of OECD guideline 407.•The clinical signs included death and acute cardiac distress in 2 out of 5 rats in the highest dose group (15mg/kg b.w.).•Moniliformin reduced the phagocytic activity of the rat neutrophils indicating an adverse effect on innate immunity.•Excretion of moniliformin into urine was rapid, with no indication of accumulation post-exposure.•Based on our findings, we suggest a LOAEL of 3mg/kg b.w.

We evaluated total and inorganic arsenic levels in long grain rice and rice based baby foods on Finnish market. Inorganic arsenic was analysed with an HPLC-ICP-MS system. The total arsenic concentration was determined with an ICP-MS method. In this study, the inorganic arsenic levels in long grain rice varied from 0.09 to 0.28mg/kg (n=8) and the total arsenic levels from 0.11 to 0.65mg/kg. There was a good correlation between the total and inorganic arsenic levels in long grain rice at a confidence level of 95%. The total arsenic levels of rice-based baby foods were in the range 0.02 - 0.29mg/kg (n=10), however, the level of inorganic arsenic could only be quantitated in four samples, on average they were 0.11mg/kg. Our estimation of inorganic arsenic intake from long grain rice and rice-based baby food in Finland indicate that in every age group the intake is close to the lowest BMDL0.1 value 0.3μg/kg bw/day set by EFSA. According to our data, the intake of inorganic arsenic should be more extensively evaluated.

The determination of PAH metabolites in fish bile is performed to assess the PAH contamination in fish for environmental monitoring. A growing number of laboratories in Europe use this parameter for national monitoring. However, an international intercomparison between laboratories has not taken place in the last years. Therefore the determination of the PAH metabolite 1-hydroxypyrene was tested in a collaborative trial performed by 10 laboratories from eight countries. Five samples of naturally contaminated fish bile covering different concentration levels were distributed among the participants. The present study was open for different methods: GS/MS, HPLC-fluorescence (HPLC-F), fixed wavelength fluorescence (FWF), and synchronous fluorescence spectrometry (SFS), respectively. The results suggest that all four methods under investigation are suitable for screening purposes, but only three methods have produced comparable results which could be used for a common monitoring database: GC/MS, HPLC-F, and SFS (with conversion factor). Most z-scores were within the acceptance criteria of ±2. The comparability of GC/MS, HPLC-F, and SFS results should be further improved because SFS is a widely used method with a great potential for monitoring. The present study contributes to the quality assurance of results for European marine monitoring.

Moniliformin is a Fusarium mycotoxin highly prevalent in grains and grain-based products worldwide. In this study, the acute oral toxicity of moniliformin was assessed in Sprague–Dawley male rats according to OECD Guideline 423 with a single-dose exposure. Clinical observations and histopathological changes were recorded together with the excretion of moniliformin via urine and feces, utilizing a novel liquid chromatography–mass spectrometry method.According to our study, moniliformin is acutely toxic to rats with a rather narrow range of toxicity. Moniliformin can be classified into category 2 (LD50 cut-off value 25mg/kg b.w.), according to the Globally Harmonized System for the classification of chemicals. The clinical observations included muscular weakness, respiratory distress and heart muscle damage. Pathological findings confirmed that heart is the main target tissue of acute moniliformin toxicity. A significant proportion (about 38%) of the administered moniliformin was rapidly excreted in urine in less than 6h. However, the toxicokinetics of the majority of the administered dose still requires clarification, as the total excretion was only close to 42%. Considering the worldwide occurrence of moniliformin together with its high acute toxicity, research into the subchronic toxicity is of vital importance to identify the possible risk in human/animal health.

Some mutations in mitochondrial DNA (mtDNA) causing a number of neuromuscular diseases are suggested to arise spontaneously during the life of an individual. To substantiate the extent and the rate of these somatic mutations, mtDNA specimens from post-mortem human heart muscles of subjects in differing age groups were hydrolyzed. 8-Hydroxy-deoxyguanosine (8-OH-dG), a hydroxyl-radical adduct of deoxyguanosine, in mtDNA, was quantitatively determined using a micro high-performance liquid chromatography/mass spectrometry system. In each specimen, the mtDNA with a 7.4 kilo base-pair deletion was quantified by the kinetic polymerase chain reaction method. In association with age, the 8-OH-dG content accumulated exponentially up to 1.5 % with a correlative increase in the content of the deleted mtDNA up to 7 %. Clear correlation between the 8-OH-dG content in mtDNA and the population of mtDNA with a deletion (r = 0.93, P < 0.01) gives insight into the mechanism for the generation of a large deletion. These results indicate that accumulation of somatically acquired oxygen damage together with age-associated mutations in mtDNA which lead to bioenergetic deficiency and the heart muscle weakness are inevitable in human life.

DNA or protein adducts are reaction products of endogenous or exogenous chemicals and cellular macromolecules. Adducts are useful in toxicological studies and/or human biomonitoring exercises. In particular, DNA damage provides invaluable information for risk analysis. Second, metabolites or conjugates can be regarded as markers of phase II reactions though they may not give accurate information about the levels of reactive and damage-provoking reactive compounds or intermediates. Electrophiles are often short-lived molecules and therefore difficult to monitor. In contrast, adducts are often chemically stable, though their levels in biological samples are low, which makes their detection challenging. The assay of adducts is similar to the analysis of any other trace organic molecule, i.e. problems with the matrix and small amounts of analytes in samples. The (32)P-postlabelling assay is a specific method for DNA adducts but immunochemical and fluorescence-based methods have been developed which can detect adducts linked to both DNA and protein. Tandem mass spectrometry, particularly if combined with ultrahigh-performance liquid chromatography, is currently the recommended detection technique; however investigators are striving to develop novel ways to achieve greater sensitivity. Standards are a prerequisite in adduct analysis, but unfortunately they are seldom commercially available.

The performance of vector (3D) filtering of color and multichannel images in discrete cosine transform (DCT) domain is studied. It is shown that the vector filtering offers some advantages compared to the component-wise (2D) filtering. Moreover, it is shown through experiments that the DCT based processing can be more efficient than advanced nonlinear vector filters. As noise models, both additive noise and mixed additive and impulsive noise are used. Finally, the applicability of the DCT based filtering is studied when the noise is multiplicative; and a convenient way to reduce multiplicative noise to additive noise is proposed.

The 2‐oxoglutarate (2OG)‐ and Fe2+‐dependent dioxygenase AlkB couples the demethylation of modified DNA to the decarboxylation of 2OG. Extensive crystallographic analyses have shown no evidence of significant structural differences between complexes binding either 2OG or succinate. By using nuclear magnetic resonance spectroscopy, we have shown that the AlkB–succinate and AlkB–2OG complexes have significantly different dynamic properties in solution. 2OG makes the necessary contacts between the metal site and the large β‐sheet to maintain a fully folded conformation. Oxidative decarboxylation of 2OG to succinate leads to weakening of a main contact with the large β‐sheet, resulting in an enhanced dynamic state. These conformational fluctuations allow for the replacement of succinate in the central core of the protein and probably contribute to the effective release of unmethylated DNA. We also propose that the inherent dynamics of the co‐product complex and the subsequent increased molecular ordering of the co‐substrate complex have a role in DNA damage recognition.

The performance of recursive and nonrecursive soft mor- phological filters is compared by studying their impulse removal capabilities. For this, the breakdown probabili- ties of the filters are applied. Formulas for the breakdown probabilities of basic nonrecursive operations are briefly reviewed, and a method for the calculation of the break- down probabilities of basic recursive soft morphological filters is given. Also, comparison of nonrecursive and re- cursive soft morphological filters is done and examples are given.

A common feature of multichannel images of different origin and spectral bands is that they exhibit a high degree of mutual correlation of their components. At the same time, information content and noise properties in component images are more or less different. For the noise removal from such images, two general approaches are possible. The first one assumes component-wise (2-D) filtering; the second approach presumes vector (3-D) processing. Component wise filtering is, in general, simpler whilst 3-D processing that is able to take into account the inter-channel (inter-band) correlation of data is potentially more efficient.