The transferability of diatoms to clothing and the methods appropriate for their collection and analysis in forensic geoscience

AbstractForensic geoscience is concerned with the analysis of geological materials in order to compare and exclude environmental samples from a common source, or to identify an unknown provenance in a criminal investigation. Diatom analysis is currently an underused technique within the forensic geoscience approach, which has the potential to provide an independent ecological assessment of trace evidence. This study presents empirical data to provide a preliminary evidence base in order to be able to understand the nature of diatom transfers to items of clothing, and the collection of transferred diatom trace evidence from a range of environments under experimental conditions. Three diatom extraction methods were tested on clothing that had been in contact with soil and water sites: rinsing in water (RW), rinsing in ethanol (RE), and submersion in H2O2 solution (H). Scanning electron microscopy (S.E.M.) analysis was undertaken in order to examine the degree of diatom retention on treated clothing samples. The total diatom yield and species richness data was recorded from each experimental sample in order to compare the efficacy of each method in collecting a representative sample for analysis. Similarity was explored using correspondence analysis. The results highlight the efficiency of H2O2 submersion in consistently extracting high diatom counts with representative species from clothing exposed to both aquatic and terrestrial sites. This is corroborated by S.E.M. analysis. This paper provides an important empirical evidence base for both establishing that diatoms do indeed transfer to clothing under forensic conditions in a range of environments, and in identifying that H2O2 extraction is the most efficient technique for the optimal collection of comparative samples. There is therefore potentially great value in collecting and analysing diatom components of geoforensic samples in order to aid in forensic investigation

Insulitis in human type 1 diabetes: lessons from an enigmatic lesion

This is the author accepted manuscript. The final version is available on open access from Oxford University Press via the DOI in this recordData availability: Relevant images from the Exeter Archival Diabetes Biobank are available from the author on reasonable request and many can be viewed online at: www.pancreatlas.orgType 1 diabetes is caused by a deficiency of insulin secretion which has been considered traditionally as the outcome of a precipitous decline in the viability of β-cells in the islets of Langerhans, brought about by autoimmune-mediated attack. Consistent with this, various classes of lymphocyte, as well as cells of the innate immune system have been found in association with islets during disease progression. However, analysis of human pancreas from subjects with type 1 diabetes has revealed that insulitis is often less intense than in equivalent animal models of the disease and can affect many fewer islets than expected, at disease onset. This is especially true in subjects developing type 1 diabetes in, or beyond, their teenage years. Such studies imply that both the phenotype and the number of immune cells present within insulitic lesions can vary among individuals in an age-dependent manner. Additionally, the influent lymphocytes are often mainly arrayed peripherally around islets rather than gaining direct access to the endocrine cell core. Thus, insulitis remains an enigmatic phenomenon in human pancreas and this review seeks to explore the current understanding of its likely role in the progression of type 1 diabetes.Medical Research Council (MRC)Diabetes UKJDRFEFSDInnovative Medicines Initiative 2 Joint UndertakingEuropean Union Horizon 2020European Federation of Pharmaceutical Industries and Associations (EFPIA)Leona M. and Harry B. Helmsley Charitable TrustNational Institute for Health and Care Research (NIHR

Spatiotemporal dynamics of insulitis in human Type 1 diabetes

This is the final version of the article. Available from the publisher via the DOI in this record.Type 1 diabetes (T1D) is an auto-immune disease characterised by the selective destruction of the insulin secreting beta cells in the pancreas during an inflammatory phase known as insulitis. Patients with T1D are typically dependent on the administration of externally provided insulin in order to manage blood glucose levels. Whilst technological developments have significantly improved both the life expectancy and quality of life of these patients, an understanding of the mechanisms of the disease remains elusive. Animal models, such as the NOD mouse model, have been widely used to probe the process of insulitis, but there exist very few data from humans studied at disease onset. In this manuscript, we employ data from human pancreases collected close to the onset of type 1 diabetes and propose a spatio-temporal computational model for the progression of insulitis in human T1D, with particular focus on the mechanisms underlying the development of insulitis in pancreatic islets. This framework allows us to investigate how the time-course of insulitis progression is affected by altering key parameters, such as the number of the CD20+ B cells present in the inflammatory infiltrate, which has recently been proposed to influence the aggressiveness of the disease. Through the analysis of repeated simulations of our stochastic model which track the number of beta cells within an islet, we find that increased numbers of B cells in the peri-islet space lead to faster destruction of the beta cells. We also find that the balance between the degradation and repair of the basement membrane surrounding the islet is a critical component in governing the overall destruction rate of the beta cells and their remaining number. Our model provides a framework for continued and improved spatio-temporal modelling of human T1D.This work was generously supported by the Wellcome Trust Institutional Strategic Support Award (WT105618MA). KT gratefully acknowledges the financial support of the EPSRC via grant EP/N014391/1. We are also pleased to acknowledge financial support from the European Unions Seventh Framework Programme PEVNET [FP7/2007-2013] under grant agreement number 261441 to NM. The participants of the PEVNET consortium are described at http://www.uta.fi/med/pevnet/ publications.html. Additional support was from a JDRF Career Development Award (5-CDA-2014-221-A-N) to SR and project grant 15/0005156 from Diabetes UK (to NM and SR)

Freshwater diatom persistence on clothing II: Further analysis of species assemblage dynamics over investigative timescales

Diatoms are a useful form of environmental trace evidence, yielding a circumstantial link between persons and scenes of forensic interest. A developing empirical research base has sought to understand those factors affecting the transfer and persistence of freshwater diatoms on clothing and footwear surfaces. Although an initial study has demonstrated that diatoms can persist on clothing following weeks of wear, no previous research has explored the temporal dynamics of a persistent species assemblage over timescales pertinent to forensic investigations. This study therefore aimed to determine if: (1) valve morphology (size and shape) influences diatom persistence, (2) the relative abundance of taxa within an assemblage affects retention, and (3) a persistent diatom assemblage retrieved from clothing after one month can reliably be compared to the site of initial transfer. To build on previous research findings which highlighted the impact of substrate and environmental seasonality on diatom transfer and persistence, here, nine clothing materials were tested in spring before a seasonal comparison in the winter. Fabric swatches were immersed in a freshwater river, worn attached to clothing, and subsamples retrieved at regular intervals (hours, days, weeks) up to one month post-immersion. Diatoms were extracted using a H2O2 technique and analysed via microscopy. The results indicated that smaller diatoms (< 10 µm) are retained in significantly greater abundance, with no statistically significant difference between centric and pennate diatom loss over time. Although a persistent species assemblage was relatively stable over the one month of wear, significant differences were identified between clothing substrate in the spring and between the seasonal samples. The most abundant environmental taxa were consistently identified in the forensic samples, with greater variability attributed to the retention of relatively less common species. The findings suggest that, despite a loss in the abundance and species-richness of diatoms retrieved from clothing over time, a persistent assemblage may provide a useful circumstantial link to the site of initial transfer. The complex relationships between clothing type, environmental seasonality, and time since wear on retention, emphasise the need for diatom trace evidence to be carefully interpreted within an exclusionary framework, and the significance of any casework findings to be determined with reference to empirical evidence bases

The transcription factor STAT6 plays a critical role in promoting beta cell viability and is depleted in islets of individuals with type 1 diabetes

This is the final version. Available on open access from Springer Verlag via the DOI in this recordAims/hypothesis: In type 1 diabetes, selective beta cell loss occurs within the inflamed milieu of insulitic islets. This milieu is generated via the enhanced secretion of proinflammatory cytokines and by the loss of anti-inflammatory molecules such as IL-4 and IL-13. While the actions of proinflammatory cytokines have been well-studied in beta cells, the effects of their anti-inflammatory counterparts have received relatively little attention and we have addressed this. Methods: Clonal beta cells, isolated human islets and pancreas sections from control individuals and those with type 1 diabetes were employed. Gene expression was measured using targeted gene arrays and by quantitative RT-PCR. Protein expression was monitored in cell extracts by western blotting and in tissue sections by immunocytochemistry. Target proteins were knocked down selectively with interference RNA. Results: Cytoprotection achieved with IL-4 and IL-13 is mediated by the early activation of signal transducer and activator of transcription 6 (STAT6) in beta cells, leading to the upregulation of anti-apoptotic proteins, including myeloid leukaemia-1 (MCL-1) and B cell lymphoma-extra large (BCLXL). We also report the induction of signal regulatory protein-α (SIRPα), and find that knockdown of SIRPα is associated with reduced beta cell viability. These anti-apoptotic proteins and their attendant cytoprotective effects are lost following siRNA-mediated knockdown of STAT6 in beta cells. Importantly, analysis of human pancreas sections revealed that STAT6 is markedly depleted in the beta cells of individuals with type 1 diabetes, implying the loss of cytoprotective responses. Conclusions/interpretation: Selective loss of STAT6 may contribute to beta cell demise during the progression of type 1 diabetes.Diabetes UKJDR

Freshwater diatom persistence on clothing I: A quantitative assessment of trace evidence dynamics over time

Freshwater diatoms offer valuable circumstantial forensic indicators, with a growing empirical research base aiming to identify and understand some of the spatial and temporal factors affecting their validity as trace evidence. Previous studies demonstrated that recipient surface characteristics, environmental variability, and individual species traits influence the initial transfer of freshwater diatoms to clothing. However, no previous research has sought to consider the impact of these and other variables on the persistence of transferred diatoms over investigative timescales. Therefore, this study aimed to identify and explore diatom retention dynamics on clothing following wear over time (hours to weeks). A series of experiments were designed to examine the impact of clothing material, seasonality, and time since wear (persistence interval) on the total number and species-richness of diatoms recovered and their relative retention (%) over time. Nine clothing swatches were immersed in a freshwater environment and then worn for one month in the spring. Subsamples were retrieved at regular intervals (e.g. 30 mins, 1 h, 8 h, 24 h) up to one month, diatoms were extracted using a H2O2 method, and examined microscopically. Three clothing materials were subject to the same experiment in the winter to generate a seasonal comparison. The results broadly identified three stages of diatom persistence on clothing - rapid initial loss, variable intermediate decay, and sustained long-term presence. Clothing material significantly impacted the number of diatoms recovered and retention dynamics over time, with complex interactions identified with seasonality. Although fewer diatoms were recovered in the winter, overall retention trends were consistent at the different times of year. The findings demonstrate that diatoms can be recovered from clothing, even weeks or months after an initial transfer, yielding a useful environmental trace indicator for forensic reconstructions over investigative timescales. The impact of clothing material and seasonality on persistence identified cotton, acrylic, and viscose clothing as the most reliable temporal repository of diatom trace evidence, with a more abundant forensic assemblage available for forensic comparisons in the spring

Cellular stressors may alter islet hormone cell proportions by moderation of alternative splicing patterns

Changes to islet cell identity in response to type 2 diabetes (T2D) have been reported in rodent models, but are less well characterized in humans. We assessed the effects of aspects of the diabetic microenvironment on hormone staining, total gene expression, splicing regulation and the alternative splicing patterns of key genes in EndoC-βH1 human beta cells. Genes encoding islet hormones [somatostatin (SST), insulin (INS), Glucagon (GCG)], differentiation markers [Forkhead box O1 (FOXO1), Paired box 6, SRY box 9, NK6 Homeobox 1, NK6 Homeobox 2] and cell stress markers (DNA damage inducible transcript 3, FOXO1) were dysregulated in stressed EndoC-βH1 cells, as were some serine arginine rich splicing factor splicing activator and heterogeneous ribonucleoprotein particle inhibitor genes. Whole transcriptome analysis of primary T2D islets and matched controls demonstrated dysregulated splicing for ~25% of splicing events, of which genes themselves involved in messenger ribonucleic acid processing and regulation of gene expression comprised the largest group. Approximately 5% of EndoC-βH1 cells exposed to these factors gained SST positivity in vitro. An increased area of SST staining was also observed ex vivo in pancreas sections recovered at autopsy from donors with type 1 diabetes (T1D) or T2D (9.3% for T1D and 3% for T2D, respectively compared with 1% in controls). Removal of the stressful stimulus or treatment with the AKT Serine/Threonine kinase inhibitor SH-6 restored splicing factor expression and reversed both hormone staining effects and patterns of gene expression. This suggests that reversible changes in hormone expression may occur during exposure to diabetomimetic cellular stressors, which may be mediated by changes in splicing regulation.This article is freely available via Open Access. Click on the Publisher URL to access it via the publisher's site.Animal Free Research UK (postgraduate studentship to L.W.H.); JDRF Career Development Award (5-CDA-2014-221-A-N to S.J.R., 5-SRA-2018-557-Q-R); Network for Pancreatic Organ donors with Diabetes (RRID:SCR_014641); The Leona M. & Harry B. Helmsley Charitable Trust (2018PG-T1D053).published version, accepted version (12 month embargo), submitted versio

Automatic Segmentation of Fluorescence Lifetime Microscopy Images of Cells Using Multi-Resolution Community Detection

We have developed an automatic method for segmenting fluorescence lifetime (FLT) imaging microscopy (FLIM) images of cells inspired by a multi-resolution community detection (MCD) based network segmentation method. The image processing problem is framed as identifying segments with respective average FLTs against a background in FLIM images. The proposed method segments a FLIM image for a given resolution of the network composed using image pixels as the nodes and similarity between the pixels as the edges. In the resulting segmentation, low network resolution leads to larger segments and high network resolution leads to smaller segments. Further, the mean-square error (MSE) in estimating the FLT segments in a FLIM image using the proposed method was found to be consistently decreasing with increasing resolution of the corresponding network. The proposed MCD method outperformed a popular spectral clustering based method in performing FLIM image segmentation. The spectral segmentation method introduced noisy segments in its output at high resolution. It was unable to offer a consistent decrease in MSE with increasing resolution.Comment: 21 pages, 6 figure

Pedagogias trans/plurilíngues: Uma Multietnografia

Trans/plurilingual theory and pedagogies have been generating extensive attention within global English language teaching and teacher education. This article responds to the burgeoning area of trans/plurilingual pedagogies, outlining diverse pedagogical practices and perspectives from a group of English language educators at a large, cosmopolitan Canadian university. Considering recent assertions that there are onto-epistemological differences between multilingualism, plurilingualism, and translingualism, this article looks to demonstrate how (or even if) these differences manifest in the pedagogical practices of diverse faculty within the same language teaching and teacher education programs. Drawing on multiethnographic data, this article concludes with a discussion of the potential and limitations of critical pedagogies, the affordances of multiethnography as an accessible methodology for use by researchers and pedagogues, and a call for greater bi-directional knowledge flow between language researchers and classroom instructors.A teoria e as pedagogias trans/plurilíngues têm gerado uma atenção extensiva dentro do ensino global da língua inglesa e formação de professores. Este artigo responde à crescente área de pedagogias trans/plurilíngues, esboçando diversas práticas e perspectivas pedagógicas de um grupo de educadores de língua inglesa em uma grande e cosmopolita universidade canadense. Tomando como ponto de partida a afirmação recente de que existem diferenças epistemológicas entre multilinguismo, plurilinguismo e translinguismo, este artigo procura demonstrar como (ou mesmo se) essas diferenças se manifestam nas práticas pedagógicas de diversos docentes dentro de um mesmo programa de ensino de línguas e de formação de professores. Este artigo conclui com uma discussão sobre o potencial e as limitações das pedagogias "críticas", as possibilidades da multietnografia como uma metodologia acessível para uso por pesquisadores e pedagogos, e um apelo para um maior fluxo de conhecimento bidirecional entre pesquisadores de idiomas e instrutores de sala de aula