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Population declines and demographic changes of Chinook salmon (Oncorhynchus tshawytscha), have been documented throughout this species’ range, though information on natural and anthropogenic mechanisms related to these changes are not fully understood. To provide insights into marine behaviors and survival of Chinook salmon, 40 pop-up satellite archival tags (PSATs), that collected environmental data, were attached to large (69–100 cm FL) Chinook salmon caught in the marine waters of Cook Inlet, Alaska. PSATs provided evidence of predation on tagged Chinook salmon by ectothermic and unconfirmed predators, and provided valuable information about the migratory characteristics and occupied depths and temperatures of this species while occupying Cook Inlet and the Gulf of Alaska. The results from this study suggest that late-marine mortality of Chinook salmon of a variety of stock-origins by apex predators is more common in Cook Inlet than previously thought, and may be used to improve our understanding this species’ population dynamics. Furthermore, results from this study adds to the existing knowledge of marine habitat use by Chinook Salmon and may be useful in assessing the vulnerability and interactions between this species and anthropogenic activities.

open access article Cells are continually exposed to agents arising from the internal and external environments, which may damage DNA. This damage can cause aberrant cell function, and therefore DNA damage may play a critical role in the development of, conceivably, all major human diseases, e.g., cancer, neurodegenerative and cardiovascular disease, and aging. Single-cell gel electrophoresis (i.e., the comet assay) is one of the most common and sensitive methods to study the formation and repair of a wide range of types of DNA damage (e.g., single- and double-strand breaks, alkali-labile sites, DNA-DNA crosslinks, and, in combination with certain repair enzymes, oxidized purines, and pyrimidines), in both in vitro and in vivo systems. However, the low sample throughput of the conventional assay and laborious sample workup are limiting factors to its widest possible application. With the "scoring" of comets increasingly automated, the limitation is now the ability to process significant numbers of comet slides. Here, a high-throughput (HTP) variant of the comet assay (HTP comet assay) has been developed, which significantly increases the number of samples analyzed, decreases assay run time, the number of individual slide manipulations, reagent requirements, and risk of physical damage to the gels. Furthermore, the footprint of the electrophoresis tank is significantly decreased due to the vertical orientation of the slides and integral cooling. Also reported here is a novel approach to chilling comet assay slides, which conveniently and efficiently facilitates the solidification of the comet gels. Here, the application of these devices to representative comet assay methods has been described. These simple innovations greatly support the use of the comet assay and its application to areas of study such as exposure biology, ecotoxicology, biomonitoring, toxicity screening/testing, together with understanding pathogenesis.

A previously unrecognized specimen ofProtoichthyosaurus prostaxalis, LEICT G142.1991, from the Lower Jurassic of Barrow-upon-Soar, Leicestershire, UK, includes an almost complete three-dimensional skull that provides new information on the configuration of the skull roof. The position of the pineal foramen (between the frontals and the parietals) and an elongated internasal foramen in a depression along the midline of the nasals are clearly shown. The maxilla makes up a significant portion of the external naris ventral margin, an unusual character for the genus/species. This reflects intraspecific variation, not evidence of a new taxon. The specimen enables comparisons of skull roof morphology withIchthyosaurusandStenopterygius, two common Early Jurassic taxa. In particular, the position of the pineal foramen is similar toStenopterygius, but distinguishesProtoichthyosaurusfromIchthyosaurus. The lack of a frontal–prefrontal contact and the posteriorly wide nasals distinguishesProtoichthyosaurusfromStenopterygius. We also present a revised reconstruction of the skull roof morphology ofIchthyosaurus. Three additional specimens ofProtoichthyosaurusare referred to the genus: another partial skull, referred toP. prostaxalis, and two isolated forefins, identified by their unique morphology.

Throughout the northern extent of its range, Chinook salmon (Oncorhynchus tshawytscha) adult returns have been in decline for over a decade, leading to severe harvest restrictions on subsistence, commercial, and recreational fisheries. In addition to these overall declines in abundance, changes in size structure and age structure, including a proportional decrease of older age classes returning to spawn, suggest that late-stage marine mortality for this species may be more frequent than currently assumed. To examine this late-stage mortality hypothesis, we examined diagnostic evidence of predation on large (57–100 cm fork length) Chinook salmon (n = 33) from depth, temperature, and light records collected during recent satellite tagging research. Satellite tags provided evidence of predation on tagged Chinook salmon by salmon sharks (Lamna ditropis) (n = 14), marine mammals (n = 2), ectothermic fish(es) (n = 3), and unidentified predators (n = 5) in the Bering Sea and Gulf of Alaska. High mortality rates in this study suggest that fisheries scientists should consider that late-stage mortality by marine apex predators may be shaping this species’ abundance and demographics.

Oxidatively generated damage to DNA has been implicated in the pathogenesis of a wide variety of diseases. Increasingly, interest is also focusing upon the effects of damage to the other nucleic acids, RNA and the (2′-deoxy-)ribonucleotide pools, and evidence is growing that these too may have an important role in disease. LC-MS/MS has the ability to provide absolute quantification of specific biomarkers, such as 8-oxo-7,8-dihydro-2′-deoxyGuo (8-oxodG), in both nuclear and mitochondrial DNA, and 8-oxoGuo in RNA. However, significant quantities of tissue are needed, limiting its use in human biomonitoring studies. In contrast, the comet assay requires much less material, and as little as 5 μL of blood may be used, offering a minimally invasive means of assessing oxidative stress in vivo, but this is restricted to nuclear DNA damage only. Urine is an ideal matrix in which to non-invasively study nucleic acid-derived biomarkers of oxidative stress, and considerable progress has been made towards robustly validating these measurements, not least through the efforts of the European Standards Committee on Urinary (DNA) Lesion Analysis. For urine, LC-MS/MS is considered the gold standard approach, and although there have been improvements to the ELISA methodology, this is largely limited to 8-oxodG. Emerging DNA adductomics approaches, which either comprehensively assess the totality of adducts in DNA, or map DNA damage across the nuclear and mitochondrial genomes, offer the potential to considerably advance our understanding of the mechanistic role of oxidatively damaged nucleic acids in disease., Graphical abstract Image 1, Highlights • Oxidatively damaged nucleic acids are implicated in the pathogenesis of disease. • LC-MS/MS, comet assay and ELISA are often used to study oxidatively damaged DNA. • Urinary oxidatively damaged nucleic acids non-invasively reflect oxidative stress. • DNA adductomics will aid understanding the role of ROS damaged DNA in disease.

Pulp canal calcification is 1 of the possible outcomes after certain types of dental trauma. This can make endodontic treatment more challenging should it become necessary. Because of the increased degree of difficulty, sometimes procedural incidents do occur during root canal treatment. This case report demonstrates an unusual clinical presentation of a root perforation and missed canal, which had undergone calcification as a result of trauma some years earlier. A contemporary approach to treatment involved a combination of treating the biological complication of the calcified canal combined with surgical repair of the iatrogenic complication of a perforation using modern imaging techniques and materials.

SUMMARY The protein tau has been implicated in many brain disorders. In animal models, tau reduction suppresses epileptogenesis of diverse causes and ameliorates synaptic and behavioral abnormalities in various conditions associated with excessive excitation-inhibition (E/I) ratios. However, the underlying mechanisms are unknown. Global genetic ablation of tau in mice reduces the action potential (AP) firing and E/I ratio of pyramidal cells in acute cortical slices without affecting the excitability of these cells. Tau ablation reduces the excitatory inputs to inhibitory neurons, increases the excitability of these cells, and structurally alters their axon initial segments (AISs). In primary neuronal cultures subjected to prolonged overstimulation, tau ablation diminishes the homeostatic response of AISs in inhibitory neurons, promotes inhibition, and suppresses hypersynchrony. Together, these differential alterations in excitatory and inhibitory neurons help explain how tau reduction prevents network hypersynchrony and counteracts brain disorders causing abnormally increased E/I ratios., Graphical Abstract, In brief Tau reduction prevents epileptogenesis, but underlying mechanisms are uncertain. Chang et al. show that tau ablation decreases the baseline activity of excitatory neurons and modulates the intrinsic excitability and axon initial segments of inhibitory neurons, promoting network inhibition. In combination, these effects counteract network hypersynchrony and diseases causing excitation/inhibition imbalance.

The teaching of medical parasitology is facing important challenges including the need to reverse the current downward trend in the teaching status of this science reported in developed countries, despite increasing food and water parasitic outbreaks in these countries. Moreover, the teaching of this science should be adapted to the rapidly increasing biomedical and technological achievements in our societies, so we can meet future students’ interests and expectations as well as being able to supply future work placement needs. Thus, parasitologists from different European Universities [De Montfort University, DMU, UK; and the Spanish University of San Pablo CEU (USP-CEU) and Miguel Hernández de Elche], are developing a complete on-line package for teaching and learning medical parasitology, named DMU e-Parasitology (http://parasitology.dmu.ac.uk). This novel package includes a virtual laboratory and microscope with a complete library of digitised 2D slides of parasites in clinical samples. Recently, we have been successful in using 3D super-resolution microscopy (3D Cell Explorer; Nanolive), to incorporate 3D microscopic images (multiple-viewpoint-holographic images, 96 z-stacks) of important protozoan (e.g. http://parasitology.dmu.ac.uk /learn/3D_Parasitology/Acanthamoeba_cyst_1.htm) and fungi human parasites fixed on slides, of. In contrast to images created from pre-stained clinical samples, in which structures of the parasites were indistinguishable from the background, although insight of the morphological structure of the infective forms of the parasites could be seen in the 3D z-stack images in each fixed culture samples provided. However, we believe that such images will have little applicability as a potential diagnostic tool, requiring further development. We have also created an e-learning unit on parasite cell culture (http://parasitology.dmu.ac.uk /learn/lab/parasite_cell_cultures/story_flash.html), which show all the practices and procedures to work in a parasite culture unit in conjunction with detailed information and videos of parasitologists/technicians working in real conditions with parasite cultures. In order to validate this unit, we will use a blended learning approach with final year BSc Biomedical Science students and MSc Advanced Biomedical Science at DMU that voluntarily enrol to receive formative training in these topics. This training consists of two sessions, which will be delivered in the first week of April 2019, when these volunteer students have completed the DMU e-Parasitology’s Parasite Cell Unit. The first session, mostly theoretical, will provide an overall description of how to work in a parasite culture unit in conjunction with explanatory mini-videos, in which students will be able to observe different parasites in culture conditions and specific 2D (clinical samples) and 3D (fixed culture samples) slides. Thus, students will be able to observe the morphological structures of the infectious forms of these pathogens in three formats: as culture (live and fixed) and in a human tissue sample. In the second session students will use a class II biological safety cabinet to manage human cells and perform routinely tasks such as grow, culture and count these cells. This paper will provide an overall description of these novel resources for teaching/learning parasite culture and their effectiveness for teaching these important laboratory skills to future healthcare professionals.

The maintenance of genomic integrity is essential for normal cellular functions. However, it is difficult to maintain over a lifetime in postmitotic cells such as neurons, in which DNA damage increases with age and is exacerbated by multiple neurological disorders, including Alzheimer’s disease (AD). Here we used immunohistochemical staining to detect DNA double strand breaks (DSBs), the most severe form of DNA damage, in postmortem brain tissues from patients with mild cognitive impairment (MCI) or AD and from cognitively unimpaired controls. Immunostaining for γH2AX—a post-translational histone modification that is widely used as a marker of DSBs—revealed increased proportions of γH2AX-labeled neurons and astrocytes in the hippocampus and frontal cortex of MCI and AD patients, as compared to age-matched controls. In contrast to the focal pattern associated with DSBs, some neurons and glia in humans and mice showed diffuse pan-nuclear patterns of γH2AX immunoreactivity. In mouse brains and primary neuronal cultures, such pan-nuclear γH2AX labeling could be elicited by increasing neuronal activity. To assess whether pan-nuclear γH2AX represents DSBs, we used a recently developed technology, DNA damage in situ ligation followed by proximity ligation assay, to detect close associations between γH2AX sites and free DSB ends. This assay revealed no evidence of DSBs in neurons or astrocytes with prominent pan-nuclear γH2AX labeling. These findings suggest that focal, but not pan-nuclear, increases in γH2AX immunoreactivity are associated with DSBs in brain tissue and that these distinct patterns of γH2AX formation may have different causes and consequences. We conclude that AD is associated with an accumulation of DSBs in vulnerable neuronal and glial cell populations from early stages onward. Because of the severe adverse effects this type of DNA damage can have on gene expression, chromatin stability and cellular functions, DSBs could be an important causal driver of neurodegeneration and cognitive decline in this disease. Electronic supplementary material The online version of this article (10.1186/s40478-019-0723-5) contains supplementary material, which is available to authorized users.

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. Urinary 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-oxodGuo) is a widely measured biomarker of oxidative stress. It has been commonly assumed to be a product of DNA repair, and therefore reflective of DNA oxidation. However, the source of urinary 8-oxodGuo is not understood, although potential confounding contributions from cell turnover and diet have been ruled out. Clearly it is critical to understand the precise biological origins of this important biomarker, so that the target molecule that is oxidised can be identified, and the significance of its excretion can be interpreted fully. In the present study we aimed to assess the contributions of nucleotide excision repair (NER), by both the global genome NER (GG-NER) and transcription-coupled NER (TC-NER) pathways, and sanitisation of the dGTP pool (e.g. via the activity of the MTH1 protein), on the production of 8-oxodGuo, using selected genetically-modified mice. In xeroderma pigmentosum A (XPA) mice, in which GG-NER and TC-NER are both defective, the urinary 8-oxodGuo data were unequivocal in ruling out a contribution from NER. In line with the XPA data, the production of urinary 8-oxodGuo was not affected in the xeroderma pigmentosum C mice, specifically excluding a role of the GG-NER pathway. The bulk of the literature supports the mechanism that the NER proteins are responsible for removing damage to the transcribed strand of DNA via TC-NER, and on this basis we also examined Cockayne Syndrome mice, which have a functional loss of TC-NER. These mice showed no difference in urinary 8-oxodGuo excretion, compared to wild type, demonstrating that TC-NER does not contribute to urinary 8-oxodGuo levels. These findings call into question whether genomic DNA is the primary source of urinary 8-oxodGuo, which would largely exclude it as a biomarker of DNA oxidation. The urinary 8-oxodGuo levels from the MTH1 mice (both knock-out and hMTH1-Tg) were not significantly different to the wild-type mice. We suggest that these findings are due to redundancy in the process, and that other enzymes substitute for the lack of MTH1, however the present study cannot determine whether or not the 2’-deoxyribonucleotide pool is the source of urinary 8-oxodGuo. On the basis of the above, urinary 8-oxodGuo is most accurately defined as a non-invasive biomarker of oxidative stress, derived from oxidatively generated damage to 2’-deoxyguanosine.

This is a post-peer-review, pre-copyedit version of an article published in Environmental Biology of Fishes. The final authenticated version is available online at: https://doi.org/10.1007/s10641-019-00889-0 . Chinook salmon are widely distributed in offshore waters of the North Pacific Ocean, and of great economical and subsistence importance; however, little is known about their oceanic ecology. To address this, we tagged 43 Chinook salmon Oncorhynchus tshawytscha (57–100 cm) with pop-up satellite archival tags (PSATs) in the eastern (October– December) and central Bering Sea (August) to provide insights into the oceanic movements, behavior, and thermal environment of this species. The tags retrieved data for up to 260 days and end locations of tagged Chinook salmon spanned from the central Bering Sea (n = 6), eastern Bering Sea/Aleutian Islands (n = 20), and the Gulf of Alaska (n = 6). While at liberty, Chinook salmon occupied depths ranging from 0 to 538 m and experienced a thermal environment ranging from −0.6 to 13.5 °C. Overall, mean depths of individual fish ranged from 4.5 to 127.9 m, while median depths ranged from 1.3 to 99.5 m. Although sample sizes were not even among months of the year, Chinook salmon occupied the shallowest and warmest water in May–September and the deepest and coolest water in December–March. Diel depth-specific diving behaviors of Chinook salmon were found in some tag records, but these behaviors appeared to be variable among individuals and plastic in nature within individuals. Results from this study provide insights into movement, diving behavior and the thermal environment of individual Chinook salmon which may have future application in understanding its ecology and developing strategies to further reduce incidental catch of this species.

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. Human health courses at universities are facing different challenges to provide students with real laboratory experiences due to the combination of large course cohorts with a shortage of academic staff, resources and time. Future health professionals are encouraged to have a complete understanding of human anatomy and histology as well as to have some pathology and diagnostic skills including the necessary skills to use a microscope. However, students often find learning histology challenging as they usually need to learn how to use a microscope in a limited time during a practical timetabled with several peers, in which they are also required to undertake other activities such as tissue embedding, cutting and staining. To address these factors, different web-based resources have been made available to enhance learning of anatomy and histology, however they are directed to medical students so their use by non-medical students (e.g. pharmacy, biomedical science, nursing, etc.) is limited due to their complexity. As a result, De Montfort University (DMU, UK) is leading an international project to develop an on-line package for teaching and learning biology, named DMU e-Biology, which will cover not only the foundation but also the latest scientific knowledge on human biology. This novel resource is also equipped with a Virtual Microscope and a Virtual Laboratory. The Virtual Laboratory will present different subsections with a range of units regarding biomedical techniques and equipment, which will be developed following previous successful experience from the team. Units will be highly engaging and will contain short videos of academics and/or technicians working hands-on with equipment and/or techniques shown to bring the laboratory to the student’s house. Videos will be enhanced with audio and subtitles in English and the user will be able to complete a series of voluntary self-assessments throughout each unit to enhance engagement and self-assessment by providing the user with tools to evaluate their acquisition of knowledge. A subsection will cover all the elements to perform routine histological techniques in a biomedical laboratory, including the use and practicalities of the microtome, how to perform paraffin embedding and tissue sampling, and common staining techniques such as haematoxylin & eosin (H&E) and periodic acid–Schiff stain (PAS). The histology section will be publicly available from the DMU website in 2018 here http://parasitology.dmu.ac.uk/ebiology/biologyLaboratory_units.htm. Additionally, this resource is supported by a virtual microscope in which the user will be able to explore a library of virtual histological slides from different human tissues and organs; the virtual microscope will transfer the practicalities of a microscope to study human histology. These resources will be tested with first year BSc Biomedical Science and BMedSci Medical Science students enrolled in the module Basic Anatomy and Physiology at DMU by implementing blended learning, i.e. a pedagogy that integrates e-learning resources and materials with formal teaching (lectures, workshops and practicals), as different studies have pointed out that this pedagogy can enhance self-learning and facilitate acquisition of knowledge and long-term retention of information. This paper will provide a description of these novel resources and explore their practicalities with non-medical science students.

An international innovative teaching group from different EU Universities (De Montfort University, Leicester, UK; University of San Pablo CEU, Madrid, Spain; University of Miguel Hernandez, Elche, Spain) and biomedical scientists registered by the Health and Care Professions Council (HCPC, UK) are developing a complete e-learning package in medical parasitology for undergraduate and postgraduate students that study Health Sciences. This package, named DMU e-Parasitology, is accessible through the DMU website (http://parasitology.dmu.ac.uk) and will present different modules including a virtual laboratory module for the study of traditional and novel biomedical laboratory techniques and equipment for detecting, identifying and studying human pathogens, specifically parasites. These techniques could also be potentially used to study other pathogens such as bacteria or viruses. The virtual biomedical laboratory is under development, but is available in the DMU website here: http://parasitology.dmu.ac.uk/learn/laboratory.htm. To develop this new module of the DMU e-Parasitology, we are using Storyline 360 software and the scaffolding and methods used to build the theoretical module (Peña-Fernández et al., 2017) [1]. To facilitate the navigation, study and comprehension of the final user, we have divided the virtual laboratory into a series of sub-sections that include different units; the sub-sections so far are: microscopes (with units such as the electron microscope); molecular biology (e.g. polymerase chain reaction and gel electrophoresis); biological safety cabinets and cell/parasite culture; biochemical and immunological techniques (e.g. magnetic immunoseparation); histology (e.g. microtome) and staining techniques (e.g. Kinyoun staining). The virtual laboratory units are highly interactive and present short videos of academics and/or technicians working in real conditions with the different laboratory equipment such as a thermocycler, a microtome, or a biological safety cabinet, as well as performing a specific technique such as a staining to determine pathogens. Therefore, the user of this virtual environment will receive a complete and “real” experience of the work in a biomedical laboratory. The DMU e-Parasitology package, and specifically its virtual laboratory environment, could help technicians and students across the world to learn how to work in a biomedical laboratory as well as to perform techniques to identify and diagnose human pathogens such as microsporidia or Plasmodium spp. Thus, the virtual resource is supported by a virtual library that includes a real collection of clinical slides that will provide the user with the functionality of a light and/or an immunofluorescence microscope. In conclusion, the virtual laboratory may serve as a high quality and reliable on-line environment for the learning of techniques and equipment. These resources can be used to improve the learning of undergraduate and postgraduate students of human health sciences as well as to develop CPD training. Moreover, the virtual laboratory module may impact in the teaching of laboratory techniques and skills in developing countries due to their limited resources. This communication will explore the design and development of the virtual laboratory environment that will be publicly accessible by the end of 2018.

Following the recent communication from the European Societies of Toxicology (EUROTOX) advising that toxicology training and expertise is being eroded in the European Union, we have reviewed the teaching status of this subject in all the bioscience undergraduate courses offered at De Montfort University (DMU, UK). The courses reviewed were: Biomedical Science, Health and Wellbeing in Society, Speech and Language Therapy, Medical Science, Pharmaceutical and Cosmetic Science, Forensic Science and Pharmacy. None of these courses dedicate a complete module to the study of toxicology although they teach some aspects of toxicology following the subject-specific threshold standards described by the UK Quality Assurance Agency for Higher Education. Similar results are found in other UK Universities, although a comprehensive study on the status of toxicology teaching is needed. We have not found any undergraduate courses currently offered in the UK that contained the word “toxicology” in their title. These results are in agreement with EUROTOX, which indicated that toxicology has been generally integrated into other bioscience disciplines and is mainly offered as part of a taught postgraduate degree programme in Europe. Owing to these observations, our teaching group is performing different strategies to enhance the teaching of toxicology at DMU as we consider that the learning of this science is critically important to enable future health professionals to protect human health. These strategies included the development of specialised teaching/workshop sessions in toxicology that can be easily included in any undergraduate bioscience module. Thus, during 2016/17 we collected comprehensive feedback (during an Erasmus+ mobility grant for academics) from human health students about their views on the teaching of toxicology and one of the specialised workshops in a programme that does not offer a module in toxicology (BMedSci Medical Science, DMU) and one that does (MPharm. Pharmacy, University of San Pablo CEU, Spain). A high proportion of the students consulted requested more teaching of toxicology or the introduction of more specialised toxicology in their programmes. Thus, 85% of second year BMedSci students indicated that they would like to receive more toxicology training. Also, 42.9% (57.1% neither agree nor disagree) of fourth year MPharm. students suggested the incorporation of specialised environmental toxicology workshops within their course and all of them considered the environmental toxicology training relevant to their general toxicology module. Other strategies implemented include the enhancement of research in toxicology in our university by offering final projects on these topics to undergraduate and postgraduate students, as well as completion of PhDs. Finally, DMU has recently recruited two toxicologists as academic staff, allowing us to promote the teaching/research of toxicology as well as exploring the possibility of developing postgraduate content for the teaching of toxicology. More efforts should be considered to enhance the teaching of this subject in any bioscience programme, as the current status of toxicology in the UK has been eroded.

BackgroundTargeted therapies are based on exploiting cancer-cell-specific genetic features or phenotypic traits to selectively kill cancer cells while leaving normal cells unaffected. Oxidative stress is a cancer hallmark phenotype. Given that free nucleotide pools are particularly vulnerable to oxidation, the nucleotide pool sanitising enzyme, MTH1, is potentially conditionally essential in cancer cells. However, findings from previous MTH1 studies have been contradictory, meaning the relevance of MTH1 in cancer is still to be determined. Here we ascertained the role of MTH1 specifically in lung cancer cell maintenance, and the potential of MTH1 inhibition as a targeted therapy strategy to improve lung cancer treatments.MethodUsing siRNA-mediated knockdown or small-molecule inhibition, we tested the genotoxic and cytotoxic effects of MTH1 deficiency on H23 (p53-mutated), H522 (p53-mutated) and A549 (wildtype p53) non-small cell lung cancer cell lines relative to normal MRC-5 lung fibroblasts. We also assessed if MTH1 inhibition augments current therapies.ResultsMTH1 knockdown increased levels of oxidatively damaged DNA and DNA damage signaling alterations in all lung cancer cell lines but not normal fibroblasts, despite no detectable differences in reactive oxygen species levels between any cell lines. Furthermore, MTH1 knockdown reduced H23 cell proliferation. However, unexpectedly, it did not induce apoptosis in any cell line or enhance the effects of gemcitabine, cisplatin or radiation in combination treatments. Contrastingly, TH287 and TH588 MTH1 inhibitors induced apoptosis in H23 and H522 cells, but only increased oxidative DNA damage levels in H23, indicating that they kill cells independently of DNA oxidation and seemingly via MTH1-distinct mechanisms.ConclusionsMTH1 has a NSCLC-specific p53-independent role for suppressing DNA oxidation and genomic instability, though surprisingly the basis of this may not be reactive-oxygen-species-associated oxidative stress. Despite this, overall our cell viability data indicates that targeting MTH1 will likely not be an across-the-board effective NSCLC therapeutic strategy; rather it induces non-cytotoxic DNA damage that could promote cancer heterogeneity and evolution.

Installation of hydrokinetic power-generating devices is currently being considered for the Yukon and Tanana rivers, two large and glacially turbid rivers in Alaska. We sampled downstream-migrating fish along the margins of both rivers, a middle island in the Yukon River, and mid-channel in the Tanana River in order to assess the temporal and spatial patterns of movement by resident and anadromous fishes and hence the potential for fish interactions with hydrokinetic devices. Results suggest that (1) river margins in the Yukon and Tanana rivers are primarily utilized by resident freshwater species, (2) the mid-channel is utilized by Pacific salmon Oncorhynchus spp. smolts, and (3) only Chum Salmon O. keta smolts utilize both river margin and mid-channel areas. Some species exhibited distinct peaks and trends in downstream migration timing, including Longnose Suckers Catostomus catostomus, whitefishes (Coregoninae), Arctic Grayling Thymallus arcticus, Lake Chub Couesius plumbeus, Chinook Salmon O. ...

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link The BSc Biomedical Science (BMS) programme at De Montfort University (DMU, Leicester, UK) is accredited by the Institute of Biomedical Science (IBMS). Students enrolled within this programme acquire highly sought after skills related with human health sciences to work in: pathology departments in hospitals; research institutions; biotechnology and pharmaceutical industries; and the education sector to name a few. The degree recruits a large number of students with currently around 600 students enrolled on this programme at DMU. Despite pre-entry requirements of knowledge of subjects related to human biology, biology or chemistry, we have noted that first year students require basic support in STEM subjects (biology, chemistry and mathematics) in modules such as “Basic Microbiology”, “Basic Anatomy and Physiology” and “Chemistry for the Biosciences”. This support is especially necessary for students that come from non-traditional routes such as Business and Technology Education Council (BTEC) routes. Moreover, usually topics related with microbiology and human diseases are challenging for students, often causing stress impacting their overall performance and experience. A group of BMS academics at DMU in conjunction with universities in the European Union (EU; e.g. University of San Pablo CEU, Spain) have started to design, create and develop a series of e-learning resources or units in human biology and BMS for undergraduate students that study health sciences degrees in the EU. These units are being uploaded onto the DMU web server (http://parasitology.dmu.ac.uk/) and will be only accessible for students from participating universities during the first phase of this project (2017/18 course) in which comprehensive feedback will be collected. This web server space has three sections or modules (theoretical section, virtual laboratory and microscope) in which the new e-learning resources will be preliminary accommodated. These units will be interactive and easy to follow, and will cover basic human biology (e.g. cells, cell structure), human anatomy and physiology, histology and basic microbiology, which will be embedded in a theoretical module named DMU e-Biology within the above URL link. They will include formative assessments and case studies throughout each unit. In addition, a series of practical units are being developed which describe routine practical elements in any biomedical laboratory such as laboratory materials, pipetting, molecular techniques (e.g. PCR), cell culture (e.g. use of biological safety cabinet) and histological techniques (e.g. use of microtome, staining techniques). The development of this teaching and learning resource will cover a gap in the traditional teaching and learning methods that are currently used and provided in the participating universities. The DMU e-Biology will aid to our undergraduate students to gain knowledge in human biology and microbiology by promoting self-learning. We consider that the DMU e-Biology will help overcome spatiotemporal, equipment and resource barriers. Additionally, it may help student retention as currently about a 10% of our first year students fail to continue BMS at DMU. Finally, the creation of the DMU e-Biology will also provide support to the DMU Student Retention and Attainment Strategy 2016-2020 through the DMU Student Learning Hub, which is currently under development.

Atlantic halibut (Hippoglossus hippoglossus) in a Norwegian fjord were tagged with pop-up archival transmitting (PAT) tags to investigate whether they join offshore spawning events with halibut from other regions. All fish (n = 4) remained in the fjord throughout the spawning season, suggesting that they may be reproductively segregated from other stocks.

WOS: 000319871200001, PubMed ID: 23198723, Aims: Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical- and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject- and sample collection-related variables, as potential sources of variability. Results: Chromatographic assays showed high agreement across urines from different subjects, whereas ELISAs showed far more inter-laboratory variation and generally overestimated levels, compared to the chromatographic assays. Excretion rates in timed 'spot' samples showed strong correlations with 24 h excretion (the 'gold' standard) of urinary 8-oxodG (r(p) 0.67-0.90), although the associations were weaker for 8-oxodG adjusted for creatinine or specific gravity (SG). The within-individual excretion of 8-oxodG varied only moderately between days (CV 17% for 24 h excretion and 20% for first void, creatinine-corrected samples). Innovation: This is the first comprehensive study of both human and methodological factors influencing 8-oxodG measurement, providing key information for future studies with this important biomarker. Conclusion: ELISA variability is greater than chromatographic assay variability, and cannot determine absolute levels of 8-oxodG. Use of standardized calibrants greatly improves intra-technique agreement and, for the chromatographic assays, importantly allows integration of results for pooled analyses. If 24 h samples are not feasible, creatinine- or SG-adjusted first morning samples are recommended., ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5:"Food Quality and Safety" [FOOD-CT-2005-513943]; ECNIS2, a coordination and support action within the European Union FP7 Cooperation Theme 2 Food, Agriculture, Fisheries and Biotechnologies; CISBO; Ingeborg; Leo Dannin Foundation; National Science Council, TaiwanNational Science Council of Taiwan [NSC 97-2314-B-040-015-MY3, NSC 100-2628-B-040-001-MY4]; US NIHUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [P30ES009089]; Instituto Carlos III division of the Government for Clinical Research [PI-10/00802, RD06/0045/0006]; Generalitat ValencianaGeneralitat Valenciana [ACOM/2012/238]; Swedish Council for Working Life and Social ResearchSwedish Research CouncilSwedish Research Council for Health Working Life & Welfare (Forte); TUBITAK (Technical and Scientific Research Council of Turkey)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108Y049]; Grant Agency of the Czech RepublicGrant Agency of the Czech Republic [P503/11/0084]; Sahlgrenska University Hospital, Gothenburg; UK Medical Research Council via a People Exchange Programme Research Leader Fellowship award [G1001808/98136], Some of the authors of this work were partners in, and this work was partly supported by, ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5:"Food Quality and Safety" (Contract No. FOOD-CT-2005-513943), and also ECNIS2, a coordination and support action within the European Union FP7 Cooperation Theme 2 Food, Agriculture, Fisheries and Biotechnologies.; P Moller and S Loft are supported by CISBO and the Ingeborg and Leo Dannin Foundation.; M-R Chao and C-W Hu acknowledge financial support from the National Science Council, Taiwan (Grants NSC 97-2314-B-040-015-MY3 and NSC 100-2628-B-040-001-MY4).; R Santella acknowledges the contribution of Qiao Wang, and support from US NIH P30ES009089.; G Saez and C Cerda acknowledge financial support from the Instituto Carlos III division of the Government for Clinical Research (Grants PI-10/00802 and RD06/0045/0006) and Grant ACOM/2012/238 from Generalitat Valenciana.; K Broberg, C Lindh, and M Hossain acknowledge financial support from the Swedish Council for Working Life and Social Research; H Orhan and N Senduran acknowledge financial support from TUBITAK (Technical and Scientific Research Council of Turkey), grant number 108Y049.; P Rossner, Jr. and RJ Sram acknowledge support from the Grant Agency of the Czech Republic (P503/11/0084).; L Barregard acknowledges financial support from the Sahlgrenska University Hospital, Gothenburg.; MS Cooke acknowledges support from the UK Medical Research Council via a People Exchange Programme Research Leader Fellowship award (G1001808/98136).

The axon initial segment (AIS) is a specialized neuronal subcompartment located at the beginning of the axon that is crucially involved in both the generation of action potentials and the regulation of neuronal polarity. We recently showed that prolonged neuronal depolarization produces a distal shift of the entire AIS structure away from the cell body, a change associated with a decrease in neuronal excitability. Here, we used dissociated rat hippocampal cultures, with a major focus on the dentate granule cell (DGC) population, to explore the signaling pathways underlying activity-dependent relocation of the AIS. First, a pharmacological screen of voltage-gated calcium channels (VGCCs) showed that AIS relocation is triggered by activation of L-type Cav1 VGCCs with negligible contribution from any other VGCC subtypes. Additional pharmacological analysis revealed that downstream signaling events are mediated by the calcium-sensitive phosphatase calcineurin; inhibition of calcineurin with either FK506 or cyclosporin A totally abolished both depolarization- and optogenetically-induced activity-dependent AIS relocation. Furthermore, calcineurin activation is sufficient for AIS plasticity, because expression of a constitutively active form of the phosphatase resulted in relocation of the AIS of DGCs without a depolarizing stimulus. Finally, we assessed the role of calcineurin in other forms of depolarization-induced plasticity. Neither membrane resistance changes nor spine density changes were affected by FK506 treatment, suggesting that calcineurin acts via a separate pathway to modulate AIS plasticity. Together, these results emphasize calcineurin as a vital player in the regulation of intrinsic plasticity as governed by the AIS.

In neurons, axons possess a molecularly defined and highly organised proximal region - the axon initial segment (AIS) - that is a key regulator of both electrical excitability and cellular polarity. Despite existing as a large, dense structure with specialised cytoskeletal architecture, the AIS is surprisingly plastic, with sustained alterations in neuronal activity bringing about significant alterations to its position, length or molecular composition. However, although the upstream activity-dependent signalling pathways that lead to such plasticity have begun to be elucidated, the downstream mechanisms that produce structural changes at the AIS are completely unknown. Here, we use dissociated cultures of rat hippocampus to show that two forms of AIS plasticity in dentate granule cells - long-term relocation, and more rapid shortening - are completely blocked by treatment with blebbistatin, a potent and selective myosin II ATPase inhibitor. These data establish a link between myosin II and AIS function, and suggest that myosin II's primary role at the structure may be to effect activity-dependent morphological alterations.

In neurons, axons possess a molecularly defined and highly organised proximal region – the axon initial segment (AIS) – that is a key regulator of both electrical excitability and cellular polarity. Despite existing as a large, dense structure with specialised cytoskeletal architecture, the AIS is surprisingly plastic, with sustained alterations in neuronal activity bringing about significant alterations to its position, length or molecular composition. However, although the upstream activity-dependent signalling pathways that lead to such plasticity have begun to be elucidated, the downstream mechanisms that produce structural changes at the AIS are completely unknown. Here, we use dissociated cultures of rat hippocampus to show that two forms of AIS plasticity in dentate granule cells – long-term relocation, and more rapid shortening – are completely blocked by treatment with blebbistatin, a potent and selective myosin II ATPase inhibitor. These data establish a link between myosin II and AIS function, and suggest that myosin II’s primary role at the structure may be to effect activity-dependent morphological alterations.