M-FISH analysis shows that complex chromosome aberrations induced by α-particle tracks are cumulative products of localised rearrangements

Complex chromosome aberrations are characteristically induced after exposure to low doses of densely ionising radiation, but little is understood about their formation. To address this, we irradiated human peripheral blood lymphocytes (PBL) in vitro with 0.5 Gy densely ionising α-particles (mean of 1 α-particle/cell) and analysed the chromosome aberrations produced using 24-colour M-FISH. Our data suggest that complex formation is a consequence of direct nuclear α-particle traversal and show that the likely product of illegitimate repair of damage from a single α-particle is a single complex exchange. From an assessment of the ‘cycle structure’ of each complex exchange we predict α-particle-induced damage to be repaired at specific localised sites, and complexes to be formed as cumulative products of this repair

Chromosome breakpoint distribution of damage induced in peripheral blood lymphocytes by densely ionising radiation

Purpose: To assess the chromosomal breakpoint distribution in human peripheral blood lymphocytes (PBL) after exposure to a low dose of high linear energy transfer (LET) α-particles using the technique of multiplex fluorescence in situ hybridisation (m-FISH). Materials and methods: Separated PBL were exposed in G0 to 0.5 Gy 238 Pu α-particles, stimulated to divide and harvested ~48-50 hours after exposure. Metaphase cells were assayed by m-FISH and chromosome breaks identified. The observed distribution of breaks were then compared with expected distributions of breaks, calculated on the assumption that the distribution of breaks is random with regard to either chromosome volume or chromosome surface area. Results: More breaks than expected were observed on chromosomes 2 and 11, however no particular region of either chromosome was identified as significantly contributing to this over-representation. The identification of hot or cold chromosome regions (pter,p,cen,q,qter) varied depending on whether the data were compared according to chromosome volume or surface area. Conclusions: A deviation from randomness in chromosome breakpoint distribution was observed, and this was greatest when data were compared according to the relative surface area of each individual chromosome (or region). The identification of breaks by m-FISH (i.e. more efficient observation of interchanges than intrachanges) and importance of territorial boundaries on interchange formation are thought to contribute to these differences. The significance of the observed non-random distribution of breaks on chromosomes 2 and 11 in relation to chromatin organisation is unclear

Effect of linear energy transfer (LET) on complexity of alpha-particle-induced chromosome aberrations in human CD34+ cells.

The aim of this study was to assess the relative influence of linear energy transfer (LET) of α-particles on chromosome aberration complexity in the absence of significant other track structure differences. To do this we irradiated human haemopoietic stem cells (CD34+) with The aim of this study was to assess the relative influence of linear energy transfer (LET) of α-particles of various incident LET values (110 - 152 keV/µm, with mean LETs through the cell of 119 – 182 keV/µm) at an equi-fluence of approximately 1 α-particle/cell and assayed for chromosome aberrations by m-FISH. Based on a single harvest time to collect early division mitosis , complex aberrations were observed at comparable frequencies irrespective of incident LET, however when expressed as a proportion of the total exchanges detected, their occurrence was seen to increase with increasing LET. Cycle analysis to predict theoretical DNA double strand break rejoining cycles was also carried out on all complex chromosome aberrations detected. By doing this we found that the majority of complex aberrations are formed in single non-reducible cycles that involve just 2 or 3 different chromosomes and 3 or 4 different breaks. Each non-reducible cycle is suggested to represent ‘an area’ of finite size within the nucleus where double strand break repair occurs. We suggest that local density of damage induced and proximity of independent repair areas within the interphase nucleus determine the complexity of aberration resolved in metaphase. Overall, the most likely outcome of a single nuclear traversal of a single α-particle in CD34+ cells is a single chromosome aberration per damaged cell. As the incident LET of the α-particle increases, the likelihood of this aberration being classed as complex is greater

m-FISH analysis reveals complexity of chromosome aberrations in individuals occupationally exposed with internal plutonium: A pilot study to assess the relevance of complex aberrations as biomarkers of exposure to high-LET α particles

We recently demonstrated that a significant proportion of apparently stable insertions induced after exposure to a mean of 1 α particle/cell, detected using 3-colour FISH, were part of larger unstable complexes when visualised by 24-colour FISH. Interestingly, regardless of the long-term persistence capability of the cell, the complexity of each α particle-induced complex appeared to be specific to the nuclear traversal of a single -particle. To assess whether aberrations of a similar complexity are observed in vivo and also to examine the usefulness of detecting such aberrations as a biomarker of chronic exposure to α particles, we have carried out a limited pilot study of Russian workers with large body burdens of -particle emitting plutonium (Pu). We found unstable cells containing non-transmissible complex aberrations in all of the Pu exposed subjects analysed by m-FISH. In addition, all of the complexes seen were consistent with those previously observed in vitro. Non-transmissible complex aberrations were more common than transmissible-type complexes, consistent with on-going/chronic exposure and insertions were dominant features of both type of complex. Accordingly, this preliminary study supports the proposal that aberration complexity and non-transmissibility are the major cytogenetic features of α particle exposure that could potentially be exploited as a specific indicator of chronic exposures to high-LET α particles

Taurine uptake across the human intestinal brush-border membrane is via two transporters: H+-coupled PAT1 (SLC36A1) and Na+- and Cl−-dependent TauT (SLC6A6)

Taurine is an essential amino acid in some mammals and is conditionally essential in humans. Taurine is an abundant component of meat and fish-based foods and has been used as an oral supplement in the treatment of disorders such as cystic fibrosis and hypertension. The purpose of this investigation was to identity the relative contributions of the solute trans-porters involved in taurine uptake across the luminalmembrane of human enterocytes. Distinct transport characteristics were revealed following expression of the candidate solute trans-porters in Xenopus laevis oocytes: PAT1 (SLC36A1) is a H+-coupled, pH-dependent, Na+-and Cl−-independent, low-affinity, high-capacity transporter for taurine and β-alanine; TauT (SLC6A6) is a Na+- and Cl−-dependent, high-affinity, low-capacity transporter of taurine and β-alanine; ATB0,+ (SLC6A14) is a Na+- and Cl−-dependent, high-affinity, low-capacity transporter which accepts β-alanine but not taurine. Taurine uptake across the brush-border membrane of human intestinal Caco-2 cell monolayers showed characteristics of both PAT1-and TauT-mediated transport. Under physiological conditions, Cl−-dependent TauT-mediated uptake predominates at low taurine concentrations, whereas at higher concentrations typical of diet, Cl−-independent PAT1-mediated uptake is the major absorptive mechanism. Real-tim

Markov chain aggregation and its application to rule-based modelling

Rule-based modelling allows to represent molecular interactions in a compact and natural way. The underlying molecular dynamics, by the laws of stochastic chemical kinetics, behaves as a continuous-time Markov chain. However, this Markov chain enumerates all possible reaction mixtures, rendering the analysis of the chain computationally demanding and often prohibitive in practice. We here describe how it is possible to efficiently find a smaller, aggregate chain, which preserves certain properties of the original one. Formal methods and lumpability notions are used to define algorithms for automated and efficient construction of such smaller chains (without ever constructing the original ones). We here illustrate the method on an example and we discuss the applicability of the method in the context of modelling large signalling pathways

Fuzzy integral for rule aggregation in fuzzy inference systems

The fuzzy inference system (FIS) has been tuned and re-vamped many times over and applied to numerous domains. New and improved techniques have been presented for fuzzification, implication, rule composition and defuzzification, leaving one key component relatively underrepresented, rule aggregation. Current FIS aggregation operators are relatively simple and have remained more-or-less unchanged over the years. For many problems, these simple aggregation operators produce intuitive, useful and meaningful results. However, there exists a wide class of problems for which quality aggregation requires non- additivity and exploitation of interactions between rules. Herein, we show how the fuzzy integral, a parametric non-linear aggregation operator, can be used to fill this gap. Specifically, recent advancements in extensions of the fuzzy integral to \unrestricted" fuzzy sets, i.e., subnormal and non- convex, makes this now possible. We explore the role of two extensions, the gFI and the NDFI, discuss when and where to apply these aggregations, and present efficient algorithms to approximate their solutions

ADvanced IMage Algebra (ADIMA): a novel method for depicting multiple sclerosis lesion heterogeneity, as demonstrated by quantitative MRI.

There are modest correlations between multiple sclerosis (MS) disability and white matter lesion (WML) volumes, as measured by T2-weighted (T2w) magnetic resonance imaging (MRI) scans (T2-WML). This may partly reflect pathological heterogeneity in WMLs, which is not apparent on T2w scans

Fast Ensemble Smoothing

Smoothing is essential to many oceanographic, meteorological and hydrological applications. The interval smoothing problem updates all desired states within a time interval using all available observations. The fixed-lag smoothing problem updates only a fixed number of states prior to the observation at current time. The fixed-lag smoothing problem is, in general, thought to be computationally faster than a fixed-interval smoother, and can be an appropriate approximation for long interval-smoothing problems. In this paper, we use an ensemble-based approach to fixed-interval and fixed-lag smoothing, and synthesize two algorithms. The first algorithm produces a linear time solution to the interval smoothing problem with a fixed factor, and the second one produces a fixed-lag solution that is independent of the lag length. Identical-twin experiments conducted with the Lorenz-95 model show that for lag lengths approximately equal to the error doubling time, or for long intervals the proposed methods can provide significant computational savings. These results suggest that ensemble methods yield both fixed-interval and fixed-lag smoothing solutions that cost little additional effort over filtering and model propagation, in the sense that in practical ensemble application the additional increment is a small fraction of either filtering or model propagation costs. We also show that fixed-interval smoothing can perform as fast as fixed-lag smoothing and may be advantageous when memory is not an issue