ALBERT

Description: We prove a spectral flow formula for one-parameter families of Hamiltonian systems under homoclinic boundary conditions, which relates the spectral flow to the relative Maslov index of a pair of curves of Lagrangians induced by the stable and unstable subspaces, respectively. Finally, we deduce sufficient conditions for bifurcation of homoclinic trajectories of one-parameter families of non-autonomous Hamiltonian vector fields.

Description: We consider the Schur–Horn problem for normal operators in von Neumann algebras, which is the problem of characterizing the possible diagonal values of a given normal operator based on its spectral data. For normal matrices, this problem is well known to be extremely difficult, and in fact, it remains open for matrices of size greater than $3$ . We show that the infinite-dimensional version of this problem is more tractable, and establish approximate solutions for normal operators in von Neumann factors of type I $_\infty$ , II, and III. A key result is an approximation theorem that can be seen as an approximate multivariate analogue of Kadison's Carpenter Theorem.

Description: We study the rate of convergence to zero of the tail entropy of $C^\infty$ maps. We give an upper bound of this rate in terms of the growth in $k$ of the derivative of order $k$ and give examples showing the optimality of the established rate of convergence. We also consider the case of multimodal maps of the interval. Finally, we prove that homoclinic tangencies give rise to $C^r$ $(r\geqslant 2)$ robustly non- $h$ -expansive dynamical systems.

Description: Let ${{\mathscr {C}}}^0_{{{\mathfrak {g}}}}$ be the category of finite-dimensional integrable modules over the quantum affine algebra $U_{q}'({{\mathfrak {g}}})$ and let $R^{A_\infty }{\mbox {-}\mathrm {gmod}}$ denote the category of finite-dimensional graded modules over the quiver Hecke algebra of type $A_{\infty }$ . In this paper, we investigate the relationship between the categories ${{\mathscr {C}}}^0_{A_{N-1}^{(1)}}$ and ${{\mathscr {C}}}^0_{A_{N-1}^{(2)}}$ by constructing the generalized quantum affine Schur–Weyl duality functors ${\mathcal {F}}^{(t)}$ from $R^{A_\infty }{\mbox {-}\mathrm {gmod}}$ to ${{\mathscr {C}}}^0_{A_{N-1}^{(t)}}\ (t=1,2)$ .

Description: We present new constructions of complex and $p$ -adic Darmon points on elliptic curves over base fields of arbitrary signature. We conjecture that these points are global and present numerical evidence to support our conjecture.

Description: It has been hypothesised that positive associations between age and levels of oxidative stress-generated damage to DNA may be related to an age-dependent decline in DNA repair activity. The objective of this study was to investigate the association between age and repair activity of oxidatively damaged DNA in peripheral blood mononuclear cells (PBMCs). We isolated PBMCs from subjects aged 18–83 years, as part of a health survey of the Danish population that focussed on lifestyle factors. The level of DNA repair activity was measured as incisions on potassium bromate-damaged DNA by the comet assay. There was an inverse association between age and DNA repair activity with a 0.65% decline in activity per year from age 18 to 83 (95% confidence interval: 0.16–1.14% per year). Univariate regression analysis also indicated inverse associations between DNA repair activity and waist-hip ratio ( P 〈 0.05) and plasma concentrations of glycosylated hemoglobin ( P = 0.07). However, multivariate regression analysis only showed an inverse association between age and DNA repair activity ( P 〈 0.05), indicating that the decline in repair activity was not mediated by metabolic risk factors. In summary, the results show an inverse association between age and DNA repair activity of oxidatively damaged DNA.

Description: Exposure to traffic-related particulate matter (PM) has been associated with increased risk of lung disease, cancer and cardiovascular disease especially in elderly and overweight subjects. The proposed mechanisms involve intracellular production of reactive oxygen species (ROS), inflammation and oxidation-induced DNA damage studied mainly in young normal-weight subjects. We performed a controlled cross-over, randomised, single-blinded, repeated-measure study where 60 healthy subjects (25 males and 35 females) with age 55–83 years and body mass index above 25kg/m 2 were exposed for 5h to either particle-filtered or sham-filtered air from a busy street with number of concentrations and PM 2.5 levels of 1800/cm 3 versus 23 000/cm 3 and 3 µg/m 3 versus 24 µg/m 3 , respectively. Peripheral blood mononuclear cells (PBMCs) were collected and assayed for production of ROS with and without ex vivo exposure to nanosized carbon black as well as expression of genes related to inflammation ( chemokine (C-C motif) ligand 2 , interleukin-8 and tumour necrosis factor ), oxidative stress response ( heme oxygenase (decycling)-1 ) and DNA repair ( oxoguanine DNA glycosylase ). DNA strand breaks and oxidised purines were assayed by the alkaline comet assay. No statistically significant differences were found for any biomarker immediately after exposure to PM from urban street air although strand breaks and oxidised purines combined were significantly associated with the particle number concentration during exposure. In conclusion, 5h of controlled exposure to PM from urban traffic did not change the gene expression related to inflammation, oxidative stress or DNA repair, ROS production or oxidatively damaged DNA in PBMCs from elderly overweight human subjects.

Description: Ionising radiation causes free radical–mediated damage in cellular DNA. This damage is manifested as chromosomal aberrations and micronuclei (MN) in proliferating cells. Sesamol, present in sesame seeds, has the potential to scavenge free radicals; therefore, it can reduce radiation-induced cytogenetic damage in cells. The aim of this study was to investigate the radioprotective potential of sesamol in bone marrow cells of mice and related haematopoietic system against radiation-induced genotoxicity. A comparative study with melatonin was designed for assessing the radioprotective potential of sesamol. C57BL/6 mice were administered intraperitoneally with either sesamol or melatonin (10 and 20mg/kg body weight) 30min prior to 2-Gy whole-body irradiation (WBI) and sacrificed after 24h. Total chromosomal aberrations (TCA), MN and cell cycle analyses were performed using bone marrow cells. The comet assay was performed on bone marrow cells, splenocytes and lymphocytes. Blood was drawn to study haematological parameters. Prophylactic doses of sesamol (10 and 20mg/kg) in irradiated mice reduced TCA and micronucleated polychromatic erythrocyte frequency in bone marrow cells by 57% and 50%, respectively, in comparison with radiation-only groups. Sesamol-reduced radiation-induced apoptosis and facilitated cell proliferation. In the comet assay, sesamol (20mg/kg) treatment reduced radiation-induced comets (% DNA in tail) compared with radiation only ( P 〈 0.05). Sesamol also increased granulocyte populations in peripheral blood similar to melatonin. Overall, the radioprotective efficacy of sesamol was found to be similar to that of melatonin. Sesamol treatment also showed recovery of relative spleen weight at 24h of WBI. The results strongly suggest the radioprotective efficacy of sesamol in the haematopoietic system of mice.

Description: We introduce a new framework for the analysis of the stability of solitons for the one-dimensional Gross–Pitaevskii equation. In particular, we establish the asymptotic stability of the black soliton with zero speed.

Description: Let $k$ and $n$ be positive even integers. For a cuspidal Hecke eigenform $h$ in the Kohnen plus space of weight $k-n/2+1/2$ for $\varGamma _0(4),$ let $I_n(h)$ be the Duke–Imamo $\bar {{\text {g}}}$ lu–Ikeda lift of $h$ in the space of cusp forms of weight $k$ for ${\rm Sp}_n({{\bf{Z}}}),$ and $f$ be the primitive form of weight $2k-n$ for ${\rm SL}_2({{\bf{Z}}})$ corresponding to $h$ under the Shimura correspondence. We then express the ratio $\displaystyle {\langle I_n(h), I_n(h) \rangle / \langle h, h \rangle }$ of the period of $I_n(h)$ to that of $h$ in terms of special values of certain $L$ -functions of $f$ . This proves the conjecture proposed by Ikeda concerning the period of the Duke–Imamo $\bar {{\text {g}}}$ lu–Ikeda lift.

Description: A formula for the norm of a bilinear Schur multiplier acting from the Cartesian product $\mathcal S^2\times \mathcal S^2$ of two copies of the Hilbert–Schmidt classes into the trace class $\mathcal S^1$ is established in terms of linear Schur multipliers acting on the space $\mathcal S^\infty $ of all compact operators. Using this formula, we resolve Peller's problem on Koplienko–Neidhardt trace formulae. Namely, we prove that there exist a twice continuously differentiable function $f$ with a bounded second derivative, a self-adjoint (unbounded) operator $A$ and a self-adjoint operator $B\in \mathcal S^2$ such that \[ f(A+B)-f(A)-\left.\frac{d}{dt}(f(A+tB))\right\vert_{t=0}\notin \mathcal S^1. \]

Description: Let $\mu $ be a probability measure on $ \mathbb R^n$ with a bounded density $f$ . We prove that the marginals of $f$ on most subspaces are well-bounded. For product measures, studied recently by Rudelson and Vershynin, our results show there is a trade-off between the strength of such bounds and the probability with which they hold. Our proof rests on new affinely invariant extremal inequalities for certain averages of $f$ on the Grassmannian and affine Grassmannian. These are motivated by Lutwak's dual affine quermassintegrals for convex sets. We show that key invariance properties of the latter, due to Grinberg, extend to families of functions. The inequalities we obtain can be viewed as functional analogues of results due to Busemann–Straus, Grinberg and Schneider. As an application, we show that without any additional assumptions on $\mu $ , any marginal $\pi _E(\mu )$ , or a small perturbation thereof, satisfies a nearly optimal small-ball probability.

Description: Let $\pi :X\to \mathbb {P}^1_{\mathbb {Q}}$ be a non-singular conic bundle over $\mathbb {Q}$ having $n$ non-split fibres and denote by $N(\pi ,B)$ the cardinality of the fibres of Weil height at most $B$ that possess a rational point. Serre showed in 1990 that a direct application of the large sieve yields \[ N(\pi,B)\ll B^2(\log B)^{-n/2} \] and raised the problem of proving that this is the true order of magnitude of $N(\pi ,B)$ under the necessary assumption that there exists at least one smooth fibre with a rational point. We solve this problem for all non-singular conic bundles of rank at most 3. Our method comprises the use of Hooley neutralisers, estimating divisor sums over values of binary forms, and an application of the Rosser–Iwaniec sieve.

Description: We associate a dimer algebra $A$ to a Postnikov diagram $D$ (in a disc) corresponding to a cluster of minors in the cluster structure of the Grassmannian ${\rm Gr}(k,n)$ . We show that $A$ is isomorphic to the endomorphism algebra of a corresponding Cohen–Macaulay module $T$ over the algebra $B$ used to categorify the cluster structure of ${\rm Gr}(k,n)$ by Jensen–King–Su. It follows that $B$ can be realised as the boundary algebra of $A$ , that is, the subalgebra $eAe$ for an idempotent $e$ corresponding to the boundary of the disc. The construction and proof uses an interpretation of the diagram $D$ , with its associated plabic graph and dual quiver (with faces), as a dimer model with boundary. We also discuss the general surface case, in particular computing boundary algebras associated to the annulus.

Description: We describe a ring whose category of Cohen–Macaulay modules provides an additive categorification of the cluster algebra structure on the homogeneous coordinate ring of the Grassmannian of $k$ -planes in $n$ -space. More precisely, there is a cluster character defined on the category which maps the rigid indecomposable objects to the cluster variables and the maximal rigid objects to clusters. This is proved by showing that the quotient of this category by a single projective–injective object is Geiss–Leclerc–Schröer's category Sub $Q_k$ , which categorifies the coordinate ring of the big cell in this Grassmannian.

Description: We develop a way of seeing a complete orientable hyperbolic 4-manifold $ {\mathcal {M}}$ as an orbifold cover of a Coxeter polytope $ {\mathcal {P}} \subset \mathbb {H}^4$ that has a facet colouring. We also develop a way of finding a totally geodesic sub-manifold $ {\mathcal {N}}$ in $ {\mathcal {M}}$ , and describing the result of mutations along $ {\mathcal {N}}$ . As an application of our method, we construct an example of a complete orientable hyperbolic 4-manifold $ {\mathcal {X}}$ with a single non-toric cusp and a complete orientable hyperbolic 4-manifold ${\mathcal {Y}}$ with a single toric cusp. Both $ {\mathcal {X}}$ and $ {\mathcal {Y}}$ have twice the minimal volume among all complete orientable hyperbolic 4-manifolds.

Description: The covariogram $g_{K}$ of a convex body $K$ in $ \mathbb {R}^n$ is the function that associates to each $x\in \mathbb {R}^n$ the volume of the intersection of $K$ with $K+x$ . Determining $K$ from the knowledge of $g_K$ is known as the Covariogram Problem. It is equivalent to determining the characteristic function $1_K$ of $K$ from the modulus of its Fourier transform $\widehat {{1_K}}$ in $ \mathbb {R}^n$ , a particular instance of the Phase Retrieval Problem. We connect the Covariogram Problem to two aspects of the Fourier transform $\widehat {{1_K}}$ seen as a function in $\mathbb {C}^n$ . The first connection is with the problem of determining $K$ from the knowledge of the zero set of $\widehat {{1_K}}$ in $\mathbb {C}^n$ . To attack this problem T. Kobayashi studied the asymptotic behavior at infinity of this zero set. We obtain this asymptotic behavior assuming less regularity on $K$ and we use this result as an essential ingredient for proving that when $K$ is sufficiently smooth and in any dimension $n$ , $K$ is determined by $g_K$ in the class of sufficiently smooth bodies. The second connection is with the irreducibility of the entire function $\widehat {{1_K}}$ . This connection also shows a link between the Covariogram Problem and the Pompeiu Problem in integral geometry.

Description: The ICH S6(R1) recommendations on safety evaluation of biotherapeutics have led to uncertainty in determining what would constitute a cause for concern that would require genotoxicity testing. A Health and Environmental Sciences Institute’s Genetic Toxicology Technical Committee Workgroup was formed to review the current practice of genotoxicity assessment of peptide/protein-related biotherapeutics. There are a number of properties of peptide/protein-related biotherapeutics that distinguish such products from traditional ‘small molecule’ drugs and need to be taken into consideration when assessing whether genotoxicity testing may be warranted and if so, how to do it appropriately. Case examples were provided by participating companies and decision trees were elaborated to determine whether and when genotoxicity evaluation is needed for peptides containing natural amino acids, non-natural amino acids and other chemical entities and for unconjugated and conjugated proteins. From a scientific point of view, there is no reason for testing peptides containing exclusively natural amino acids irrespective of the manufacturing process. If non-natural amino acids, organic linkers and other non-linker chemical components have already been tested for genotoxicity, there is no need to re-evaluate them when used in different peptide/protein-related biotherapeutics. Unless the peptides have been modified to be able to enter the cells, it is generally more appropriate to evaluate the peptides containing the non-natural amino acids and other non-linker chemical moieties in vivo where the cleavage products can be formed. For linkers, it is important to determine if exposure to reactive forms are likely to occur and from which origin. When the linkers are anticipated to be potential mutagenic impurities they should be evaluated according to ICH M7. If linkers are expected to be catabolic products, it is recommended to test the entire conjugate in vivo , as this would ensure that the relevant ‘free’ linker forms stemming from in vivo catabolism are tested.

Description: Genome sequences that contain tandem repeats of guanine can form stable four-stranded structures known as G-quadruplex, or G4 DNA. While the molecular mechanisms are not fully defined, such guanine-rich loci are prone to mutagenesis and recombination. Various repair pathways function to reduce the potential for genome instability by correcting base damage and replication errors; however, it is not yet fully defined how well these processes function at G4 DNA. One frequent form of base damage occurs from cytidine deamination, resulting in deoxyuracil and UG mismatches. In duplex and single-stranded DNA, uracil bases are recognised and excised by uracil glycosylases. Here, we tested the efficiency of uracil glycosylase activity in vitro on uracil bases located directly adjacent to guanine repeats and G4 DNA. We show that uracil excision by bacterial UDG and human hUNG2 is reduced at uracils positioned directly 5' or 3' of a guanine tetrad. Control reactions using oligonucleotides disrupted for G4 formation or reaction conditions that do not favour G4 formation resulted in full uracil excision activity. Based on these in vitro results, we suggest that folding of guanine-rich DNA into G4 DNA results in a DNA conformation that is resistant to uracil glycosylase-initiated repair and this has the potential to increase the risk of instability at guanine repeats in the genome.

Description: Galectin-4 is a member of the galectin family which consists of 15 galactoside-binding proteins. Previously, galectin-4 has been shown to have a role in cancer progression and metastasis and it is found upregulated in many solid tumours, including colorectal cancer (CRC). Recently, the role in the metastatic process was suggested to be via promoting cancer cells to adhere to blood vascular endothelium. In the present study, the regulatory region of LGALS4 (galectin-4) in seven colon cell lines was investigated with respect to genetic variation that could be linked to expression levels and therefore a tumourigenic effect. Interestingly, qRT-PCR and sequencing results revealed that galectin-4 upregulation is associated with SNPs rs116896264 and rs73933062. By use of luciferase reporter- and pull-down assays, we confirmed the association between the gene upregulation and the two SNPs. Also, using pull-down assay followed by mass spectrometry, we found that the presence rs116896264 and rs73933062 is changing transcription factors binding sites. In order to assess the frequencies of the two SNPs among colon cancer patients and healthy individuals, we genotyped 75 colon cancer patients, 12 patients with adenomatous polyposis and 17 patients with ulcerative colitis and we performed data mining in the 1000 genomes databank. We found the two SNPs co-occuring in 21% of 75 CRC patients, 0 out of 12 patients of adenomatous polyposis, and 6 out of 17 patients (35%) with ulcerative colitis. Both in the patient samples and in the 1000 genomes project, the two SNPs were found to co-occur whenever present (D' = 1).

Description: The dose effect between nucleoplasmic bridges (NPB) and relatively low doses of ionising radiation remains unknown. Accordingly, this study investigated the NPB frequencies in human peripheral blood lymphocytes exposed to low-dose 60 Co -rays. Complex anomalies, including fused nuclei (FUS), horse-shoe nuclei (HS) and circular nuclei (CIR), which possibly originated from multiple NPBs, were also scored. Human peripheral blood samples were collected from three healthy males and irradiated with 0–1 and 0–0.4 Gy 60 Co -rays. A cytokinesis-block micronucleus cytome assay was then conducted to analyse NPB, PFHC (NPB plus three complex nuclear anomalies) and micronucleus (MN) in binucleated cells. All dose–response curves followed the linear model for both NPB frequency and PFHC cell frequency. The dose–response curves between NPB frequency and absorbed dose at 0–1 and 0–0.4 Gy were y = 0.0037 x + 0.0005 ( R 2 = 0.979, P 〈 0.05) and y = 0.0043 x + 0.0004 ( R 2 = 0.941, P 〈 0.05), respectively. The dose–response curves between PFHC cell frequency and absorbed dose at 0–1 and 0–0.4 Gy were y = 0.0044 x + 0.0007 ( R 2 = 0.982, P 〈 0.05) and y = 0.0059 x + 0.0005 ( R 2 = 0.969, P 〈 0.05), respectively. The statistical significance of differences between the irradiated groups (0–0.4 Gy) and background levels of NPB, PFHC and MN were also analysed. The lowest analysable doses of NPB, PFHC and MN were 0.12, 0.08 and 0.08 Gy, respectively. In conclusion, NPBs and PFHC positively correlated with the absorbed radiation at a relatively low dose.

Description: The aim of the study was to investigate how coadministration of resveratrol (RSV) at different time after the start of irradiation influences the frequency of micronuclei (MN) in reticulocytes of bone marrow and peripheral blood, and if the RSV supplementation after termination of irradiation may influence the recovery process of damaged cells. Coadministration of RSV with 1-day delay after 1 Gy irradiation significantly decreased the levels of MN in bone marrow and in peripheral blood, whereas with 1-week delay, only in bone marrow reticulocytes. Above combined treatment did not improve the process of recovery. RSV supplementation with 1-day delay relatively to 0.5 Gy irradiation, significantly decreased the frequencies of MN, especially after coadministration with 28mg/kg bw of RSV. Coadministration of RSV since eighth day did not influence the frequencies of MN compared to irradiated cells. The recovery process in the presence of RSV proceeded faster. Supplementation of RSV following initiation of irradiation is beneficial in case of irradiation with lower doses. RSV should be supplemented as soon as possible. Supplementation of RSV after termination of irradiation significantly speed up the recovery. Current results confirmed the ability of RSV to mitigate the effect of irradiation.

Description: Various naturally occurring stilbene-like compounds that are related to resveratrol (RSV) possess some of the beneficial effects of the parent molecule and provide even further benefits. Therefore, a series of methoxylated analogues of RSV were prepared with the aim of increasing antitumour and proapoptotic activity. In a previous article, we studied two methoxy-derivatives, pterostilbene (PTERO) and trimethoxystilbene (TRIMETHOXY), in which the first was formed by the substitution of two hydroxyl groups with two methoxy groups ( trans -3,5-dimethoxy-4'-hydroxystilbene) and the second was formed by the replacement of all three OH groups with methoxy groups ( trans -3,5,4'-trimethoxystilbene). Both methoxy-derivatives showed stronger antioxidant activity when compared with RSV. In the present article, we focused on the analysis of the ability of RSV and its two methoxylated derivatives to protect proliferating non-tumoural cells from the damage induced by ionising radiation (IR). First we showed that the methoxy derivatives, contrary to their parental compound, are unable to affect topoisomerase enzyme and consequently are not clastogenic per se . Second we showed that both PTERO and TRIMETHOXY more efficiently reduce the chromosome damage induced by IR. Furthermore, TRIMETHOXY, but not PTERO, causes a delay in cell proliferation, particularly in mitosis progression increasing the number of cells in metaphase at the expense of prophases and ana/telophases.

Description: α-, β- and -asarone are naturally occurring phenylpropenes that occur in different plant families, mainly in Aristolochiaceae , Acoraceae and Lauraceae. Plants containing asarones are used as flavouring ingredients in alcoholic beverages (bitters), traditional phytomedicines and the rhizome of e.g. Acorus calamus is used to prepare tea. Although α- and β-asarone show a potential in the treatment of several diseases, previous studies have shown carcinogenicity in rodents (duodenum, liver). However, the mechanism of action remained unclear. Studies on the mutagenicity of propenylic α- and β-asarone are inconsistent and data on carcinogenicity and genotoxicity of allylic -asarone are lacking completely. Thus, the present study determined the mutagenicity of the three asarone isomers using the Ames fluctuation assay with and without exogenous metabolic activation (S9 mix) in the standard Salmonella typhimurium strains TA98 and TA100. A concentration dependent increase in mutagenicity could be verified for α- and β-asarone in strain TA100 in the presence of rat liver homogenate. The side-chain epoxides of α- and β-asarone, major metabolites formed in liver microsomes, caused mutations in TA100, supporting the hypothesis that epoxidation of the side chain plays a key role in mutagenicity of the propenylic alkenylbenzenes. The allylic -asarone, not undergoing detectable side-chain epoxidation in liver microsomes, was supposed to be activated via side-chain hydroxylation and further sulphonation, a typical pathway for other allylic alkenylbenzenes like estragole or methyleugenol. However, neither y-asarone nor 1'-OH--asarone showed any mutagenic effect even in the human SULT-expressing Salmonella strains (TA100-hSULT1A1 and TA100-hSULT1C2), while 1'-OH-methyleugenol used as a positive control was mutagenic under these conditions. These results indicate that the propenylic asarones are genotoxic via metabolic formation of side-chain epoxides while the side-chain hydroxylation/sulphonation pathway is either not operative or does not lead to mutagenicity with the allylic -asarone.

Description: Prior to the downstream development of chemical substances, including pharmaceuticals and cosmetics, their influence on the genetic apparatus has to be tested. Several in vitro and in vivo assays have been developed to test for genotoxicity. In a first tier, a battery of two to three in vitro tests is recommended to cover mutagenicity, clastogenicity and aneugenicity as main endpoints. This regulatory in vitro test battery is known to have a high sensitivity, which is at the expense of the specificity. The high number of false positive in vitro results leads to excessive in vivo follow-up studies. In the case of cosmetics it may even induce the ban of the particular compound since in Europe the use of experimental animals is no longer allowed for cosmetics. In this article, an alternative approach to derisk a misleading positive Ames test is explored. Hereto we first tested the performance of five existing computational tools to predict the potential mutagenicity of a data set of 132 cosmetic compounds with a known genotoxicity profile. Furthermore, we present, as a proof-of-principle, a strategy in which a combination of computational tools and mechanistic information derived from in vitro transcriptomics analyses is used to derisk a misleading positive Ames test result. Our data shows that this strategy may represent a valuable tool in a weight-of-evidence approach to further evaluate a positive outcome in an Ames test.

Description: Pioglitazone (PTZ) is an oral antidiabetic agent whose anti-cancer properties have been described recently. Since PTZ increases the production of reactive oxygen species in mammalian cells, the aim of current study was to evaluate the cytotoxic, mutagenic and recombinogenic effects of PTZ using respectively the in vitro mitotic index assay and the in vitro mammalian cell micronucleus test in human peripheral lymphocytes, and the in vivo homozygotization assay in Aspergillus nidulans , which detects the loss of heterozygosity due to somatic recombination. Although the lowest PTZ concentrations (4–36 μM) did not show any significant rise in the micronucleus production, the higher PTZ concentration (108 μM) produced a statistically higher number of micronuclei than the negative control and significantly altered the cell-proliferation kinetics, demonstrating the mutagenic and antiproliferative effects of PTZ, respectively. The recombinogenic activity of PTZ, demonstrated here for the first time, was observed at the highest tested concentration (400 μM) through the homozygotization index rates significantly different from the negative control. Taken together, our results show that PTZ is genotoxic at a concentration higher than the therapeutic plasma concentration. This PTZ genotoxicity may be a potential benefit to its previously described antitumour activity.

Description: Environmental pollutants are complex mixtures in which metals are ubiquitous. Metal mixtures of arsenic, cadmium and lead are present in the occupational environment and generate health effects such as cardiovascular, renal and cancer diseases. Cell transformation induced by metal mixtures that depend on reactive oxygen species (ROS) generation, cell viability maintenance and avoidance of senescence was previously reported by our group. The aim of the present study was to explore the role of a Obg-like ATPase1 (OLA1) in the cell transformation of BALB/c 3T3 A31-1-1 clonal cells induced by a metal mixture (2 µM NaAsO 2 , 2 µM CdCl 2 and 5 µM Pb(C 2 H 3 O 2 ) 2 . 3H 2 O) through ROS generation. The interest in OLA1 is justified because this protein has been proposed to be a negative regulator of the cellular antioxidant response. Small interfering RNA (siRNA) was used to knockdown OLA1 before the initiation stage of the transformation assay. We evaluated (ROS) and OLA1 protein expression throughout the initiation and promotion stages of transformation. OLA1 knockdown modulated metal mixture-induced cell transformation more strongly when the metal mixture was an initiator stimulus than when it was a promoter. The ability of the metal mixture to initiate cell transformation was diminished by OLA1 knockdown, an effect that depended on intracellular ROS levels. The effect of OLA1 was synergistic with N -Acetyl- l -cysteine (NAC) co-treatment. Oxidative stress-associated transcription factors Egr1 and Smad were also down-regulated by the OLA1 knockdown, contributing to the rescue of metal mixture cell transformation.

Description: G-quadruplexes (G4) are highly stable tetra-stranded DNA secondary structures known to mediate gene regulation and to trigger genomic instability events during replication. G4 structural stability can be affected by DNA methylation and oxidation modifications; thus nutrients such as folate that have the ability to alter these processes could potentially modify the genomic occurrence of G4 elements. Hela cells were cultured in a range of folate concentrations or in the presence or absence of 5-aza-2'-deoxycytidine, a DNA-methyltransferase inhibitor. G4 structures were then quantified by immunofluorescence using an automated quantitative imaging system. G4 frequency in Hela cells and nuclei area mean were increased in 20nM folate medium compared with 2000nM folate, as well as in the presence of 5-aza-2'-deoxycytidine when compared to cells non-exposed to 5-aza-2'-deoxycytidine. These changes were exacerbated when pyridostatin, a G4 stabilising ligand, was added to the culture medium. G4 intensity in Hela cells cultured in deficient folate condition with pyridostatin was highly correlated with DNA damage as measured by H2AX immunofluorescence ( r = 0.71). This study showed for the first time that cellular G4 balance is modifiable by low folate concentrations and that these changes may occur as a consequence of DNA hypomethylation. Although the exact mechanism by which these changes occur is unclear, these findings establish the possibility that nutrients could be utilised as a tool for sustaining genome integrity by modifying G4 frequency at a cellular level.

Description: Metal oxide nanoparticles (NPs), including zinc oxide (ZnO) NPs have shown success for use as vehicles for drug delivery and targeting gene delivery in many diseases like cancer. Current anticancer chemotherapeutics fail to effectively differentiate between cancerous and normal cells. There is an urgent need to develop novel drug delivery system that can better target cancer cells while sparing normal cells and tissues. Particularly, ZnO NPs exhibit a high degree of cancer cell selectivity and induce cell death, oxidative stress, interference with the cell cycle progression and genotoxicity in cancerous cells. In this scenario, effective cellular uptake of NP seems to be crucial, which is shown to be affected by cell cycle progression. In the present study, the cytotoxic potential of ZnO NPs and the effect of different cell cycle phases on the uptake of ZnO NPs were examined in A431 cells. It is shown that the ZnO NPs led to cell death and reactive oxygen species generation and were able to induce cell cycle arrest in S and G 2 /M phase with the higher uptake in G 2 /M phase compared with other phases.

Description: Malondialdehyde (MDA), a biomarker of lipid peroxidation and oxidative stress, is a mutagenic and carcinogenic compound that can react with DNA to form several types of DNA adducts including the deoxyguanosine adduct (M 1 dG). The aim of this cross-sectional study was to evaluate the association between individual dietary and lifestyle habits and M 1 dG levels, measured in peripheral leukocytes in a large representative sample of the general population of Florence City (Italy). Selected anthropometric measurements, detailed information on dietary and lifestyle habits and blood samples were available for 313 adults of the Florence City Sample enrolled in the frame of European Prospective Investigation into Cancer and nutrition (EPIC) study. A multivariate regression analysis adjusted for selected individual characteristics possibly related to M 1 dG levels (sex, age, BMI, smoke, physical activity level, education level, total caloric intake and a Mediterranean dietary score) was performed to estimate the association between these parameters and M 1 dG levels. M 1 dG levels were significantly higher in women ( P = 0.014) and lower in moderately active or active subjects ( P = 0.037).We also found a significant inverse association with the Modified Mediterranean dietary score ( P for trend = 0.049), particularly evident for the highest categories of adherence. Our results indicate that M 1 dG levels can be modulated by selected individual characteristics such as gender, physical activity and a Mediterranean dietary pattern.

Description: Let $\varphi :X\to S$ be a morphism between smooth complex analytic spaces and let $f=0$ define a free divisor on $S$ . We prove that if the deformation space $T^1_{X/S}$ of $\varphi $ is a Cohen–Macaulay $\mathcal {O}_X$ -module of codimension 2, and all of the logarithmic vector fields for $f=0$ lift via $\varphi $ , then $f\circ \varphi =0$ defines a free divisor on $X$ ; this is generalized in several directions. Among applications we recover a result of Mond–van Straten, generalize a construction of Buchweitz–Conca, and show that a map $\varphi :\mathbb {C}^{n+1}\to \mathbb {C}^n$ with critical set of codimension 2 has a $T^1_{X/S}$ with the desired properties. Finally, if $X$ is a representation of a reductive complex algebraic group $G$ and $\varphi $ is the algebraic quotient $X\to S=X\!{/\!/} G$ with $X\!{/\!/} G$ smooth, then we describe sufficient conditions for $T^1_{X/S}$ to be Cohen–Macaulay of codimension 2. In one such case, a free divisor on $\mathbb {C}^{n+1}$ lifts under the operation of ‘castling’ to a free divisor on $\mathbb {C}^{n(n+1)}$ , partially generalizing work of Granger–Mond–Schulze on linear free divisors. We give several other examples of such representations.

Description: Let $M^n$ be a compact manifold of dimension $n$ with free $T^k$ -action. We consider collapsings of $M$ on $N=M/T^k$ such that the sectional curvature and diameter of $M$ satisfy $|K(M)|\leq a$ and $ {\rm diam}(M) 〈 d$ , and give examples of collapsings for all $k$ such that the first non-zero eigenvalue of Laplacian acting on 1-forms and 2-forms of $M$ are bounded above by $c(M)\cdot \hbox {inj}(M)^{2k}$ . Moreover, we prove that the first non-zero eigenvalue of Laplacian acting on 1-forms of all principal $T^k$ -bundle $M$ over $N$ is bounded below by $c(n,a,d,N)\cdot {\rm Vol}(M)^2$ and $c\cdot \hbox {inj}(M)^{2k}$ when $M$ collapses on $N$ .

Description: We give explicit atomic bases of arbitrary coefficient-free cluster algebras of types A and à . This entails showing that the minimal elements of the positive semiring of these cluster algebras form a linear basis over the integers for the cluster algebra.

Description: We prove that strongly F -regular and F -pure singularities satisfy Bertini-type theorems (including in the context of pairs) by building upon a framework of Cumino, Greco and Manaresi (compare with the work of Jouanolou and Spreafico). We also prove that F -injective singularities fail to satisfy even the most basic Bertini-type results.

Description: This is the second of a pair of papers on the Delta-group structure on the braid and mapping class groups of a surface. We obtain a description of the homotopy groups of these Delta-groups and generalize to an arbitrary surface the Berrick–Cohen–Wong–Wu exact sequence relating the Brunnian braid groups of the 2-sphere to its homotopy groups. We prove a similar result for Brunnian mapping class groups.

Description: We construct a geometric realization of the Khovanov–Lauda–Rouquier algebra R associated with a symmetric Borcherds–Cartan matrix A = ( a ij ) i , j I via quiver varieties. As an application, if a ii != 0 for any i I , we prove that there exists a one-to-one correspondence between Kashiwara's lower global basis (or Lusztig's canonical basis) of U A – (g) (respectively, V A ( )) and the set of isomorphism classes of indecomposable projective graded modules over R (respectively, R ).

Description: The purpose of this paper is to study the nature of quasi-invariant measures for finitely generated non-discrete subgroups of Diff ( S 1 ). For this, we apply ideas involving the closure of these groups to find out that the regularity of the measure depends on a ‘measurable version’ of well-known problems concerning stable self-intersection of Cantor sets. As applications, we prove that every d -quasiconformal probability measure for a non-solvable and non-discrete group must be absolutely continuous. Concerning singular quasi-invariant measures, it is also proved that their associated Hausdorff measures must either be zero or of infinite mass, a result contrasting with the case of dynamically defined Cantor sets and also applicable to the examples of singular stationary measures constructed by Kaimanovich and Le Prince. As a further application of our methods, a theorem of rigidity for measurable conjugations between groups as above is obtained.

Description: We study the space of period polynomials associated with modular forms of integral weight for finite-index subgroups of the modular group. For the modular group, this space is endowed with a pairing, corresponding to the Petersson inner product on modular forms via a formula of Haberland, and with an action of Hecke operators, defined algebraically by Zagier. We generalize Haberland's formula to (not necessarily cuspidal) modular forms for finite-index subgroups, and we show that it conceals two stronger formulas. We extend the action of Hecke operators to period polynomials of modular forms, we show that the pairing on period polynomials appearing in Haberland's formula is nondegenerate, and we determine the adjoints of Hecke operators with respect to it. We give a few applications for 1 ( N ): an extension of the Eichler–Shimura isomorphism to the entire space of modular forms; the determination of the relations satisfied by the even and odd parts of period polynomials associated with cusp forms, which are independent of the period relations; and an explicit formula for Fourier coefficients of Hecke eigenforms in terms of their period polynomials, generalizing the Coefficient theorem of Manin.

Description: We develop theorems which produce a multitude of hyperbolic triples for the finite classical groups. We apply these theorems to prove that every quasisimple group except Alt (5) and SL 2 (5) is a Beauville group. In particular, we settle a conjecture of Bauer, Catanese and Grunewald which asserts that all non-abelian finite quasisimple groups except for the alternating group Alt (5) are Beauville groups.

Description: Let U R d be open and convex. We prove that every (not necessarily Lipschitz or strongly) convex function f : U -〉 R can be approximated by real analytic convex functions, uniformly on all of U . We also show that C 0 -fine approximation of convex functions by smooth (or real analytic) convex functions on R d is possible in general if and only if d = 1. Nevertheless, for d ≥ 2, we give a characterization of the class of convex functions on R d which can be approximated by real analytic (or just smoother) convex functions in the C 0 -fine topology. It turns out that the possibility of performing this kind of approximation is not determined by the degree of local convexity or smoothness of the given function, but by its global geometrical behaviour. We also show that every C 1 convex and proper function on U can be approximated by C convex functions in the C 1 -fine topology, and we provide some applications of these results, concerning prescription of (sub-)differential boundary data to convex real analytic functions, and smooth surgery of convex bodies.

Description: Let $G$ be a compact connected Lie group, or more generally a path connected topological group of the homotopy type of a finite CW-complex, and let $X$ be a rational nilpotent $G$ -space. In this paper, we analyze the homotopy type of the homotopy fixed point set $X^{hG}$ , and the natural injection $k\colon X^G\hookrightarrow X^{hG}$ . We show that if $X$ is elliptic, that is, it has finite-dimensional rational homotopy and cohomology, then each path component of $X^{hG}$ is also elliptic. We also give an explicit algebraic model of the inclusion $k$ based on which we can prove, for instance, that for $G$ a torus, $\pi _* (k)$ is injective in rational homotopy but, often, far from being a rational homotopy equivalence.

Description: We employ the ergodic-theoretic machinery of scenery flows to address classical geometric measure-theoretic problems on Euclidean spaces. Our main results include a sharp version of the conical density theorem, which we show to be closely linked to rectifiability. Moreover, we show that the dimension theory of measure-theoretical porosity can be reduced back to its set-theoretic version, that Hausdorff and packing dimensions yield the same maximal dimension for porous and even mean porous measures, and that extremal measures exist and can be chosen to satisfy a generalized notion of self-similarity. These are sharp general formulations of phenomena that had been earlier found to hold in a number of special cases.

Description: A notion of tangential thickness of a manifold is introduced. An extensive calculation within the class of lens and fake lens spaces leads to a classification of such manifolds with thickness 1, 3 or 2 $k$ , for $k\geq 1$ . On the other hand, calculations of tangential thickness in terms of the dimension of the manifold and the rank of the fundamental group show very interesting and quite surprising correlations between these invariants.

Description: Extending a classical result of Widom from 1969, polynomials with small supremum norms are constructed for a large family of compact sets $\Gamma$ : their norm is at most a constant times the theoretical lower limit ${{\rm cap}}(\Gamma )^n$ , where ${{\rm cap}}(\Gamma )$ denotes logarithmic capacity. The construction is based on a discretization of the equilibrium measure, and the polynomials have the additional property that outside the given set $\Gamma$ they increase as fast as possible, namely as ${{\rm cap}}(\Gamma )^n\exp (ng_{ \overline {{{}C}}\setminus \Gamma }(z))$ , with the Green's function with pole at infinity in the exponent. This latter fact allows us to use these polynomials as building blocks in constructing Dirac delta-type polynomials around corners: if a compact set $K$ has a corner at some point $z_0$ , then Dirac delta-type polynomials (fast decreasing polynomials) peaking at $z_0$ are polynomials $P_n(z)$ with $P_n(z_0)=1$ that decrease as $|P_n(z)|\prec \exp (-n^ \beta |z-z_0|^ \gamma )$ on the set $K$ as $z$ moves away from $z_0$ . The possible $(\beta , \gamma )$ pairs are completely described in turn of the angle $\alpha \pi$ at $z_0$ ( $\beta \lt 1$ and $\gamma \ge \beta /(2- \alpha )$ or $\beta =1$ and $\gamma 〉 \beta /(2- \alpha )$ ). As application of these fast decreasing polynomials sharp Nikolskii- and Markov-type inequalities are proved for Jordan domains with corners. The paper uses distortion properties of conformal maps, potential theoretic techniques as well as the theory of weighted logarithmic potentials.

Description: We consider for every $n\in \mathbb {N}$ an algebra $\mathcal {A}_{n}$ of germs at $0\in \mathbb {R}^{n}$ of continuous real-valued functions, such that we can associate to every germ $f\in \mathcal {A}_{n}$ a (divergent) series $\mathcal {T}(f)$ with non-negative real exponents, which can be thought of as an asymptotic expansion of $f$ . We require that the $\mathbb {R}$ -algebra homomorphism $f\mapsto \mathcal {T}(f)$ be injective (quasianalyticity property). In this setting, we prove analogue results to Denef and van den Dries’ quantifier elimination theorem and Hironaka's rectilinearization theorem for subanalytic sets.

Description: Suppose that $F(x)\in \mathbb {Z}[\![x]\!]$ is a Mahler function and that $1/b$ is in the radius of convergence of $F(x)$ for an integer $b\geq 2$ . In this paper, we consider the approximation of $F(1/b)$ by algebraic numbers. In particular, we prove that $F(1/b)$ cannot be a Liouville number. If, in addition, $F(x)$ is regular, we show that $F(1/b)$ is either rational or transcendental, and in the latter case that $F(1/b)$ is an $S$ -number or a $T$ -number in Mahler's classification of real numbers.

Description: We develop techniques for computing zeta functions associated with nilpotent groups, not necessarily associative algebras, and modules, as well as Igusa-type zeta functions. At the heart of our method lies an explicit convex-geometric formula for a class of $p$ -adic integrals under non-degeneracy conditions with respect to associated Newton polytopes. Our techniques prove to be especially useful for the computation of topological zeta functions associated with algebras, resulting in the first systematic investigation of their properties.

Description: Determination of the mode of action of carcinogenic agents is an important factor in risk assessment and regulatory practice. To assess the ability of the erythrocyte-based Pig-a mutation assay to discriminate between genotoxic and non-genotoxic modes of action, the mutagenic response of Sprague Dawley rats exposed to methyl carbamate (MC) or ethyl carbamate (EC) was investigated. EC, a potent carcinogen, is believed to induce DNA damage through the formation of a DNA-reactive epoxide group, whereas the closely structurally related compound, MC, cannot form this epoxide and its weaker carcinogenic activity is thought to be secondary to inflammation and promotion of cell proliferation. The frequency of Pig-a mutant phenotype cells was monitored before, during, and after 28 consecutive days of oral gavage exposure to either MC (doses ranging from 125 to 500mg/kg/day) or EC (250mg/kg/day). Significant increases in the frequency of mutant reticulocytes were observed from Days 15 through 43, with a peak mean frequency of 19.9 x 10 –6 on Day 29 (i.e. 24.9-fold increase relative to mean vehicle control across all four sampling times). As expected, mutant erythrocyte responses lagged behind mutant reticulocyte responses, with a maximal mean frequency of 8.2 x 10 –6 on Day 43 (i.e. 16.4-fold increase). No mutagenic effects were observed with MC. A second indicator of in vivo genotoxicity, peripheral blood micronucleated reticulocytes, was also studied. This endpoint was responsive to EC (3.3-fold mean increase), but not to MC. These results support the hypothesis that genotoxicity contributes to the carcinogenicity of EC but not of MC, and illustrates the value of the Pig-a assay for discriminating between genotoxic and non-genotoxic modes of action.

Description: As part of the international Pig-a validation trials, we examined the induction of Pig-a mutant reticulocytes and red blood cells (RET CD59– and RBC CD59– , respectively) in peripheral blood of male Sprague Dawley ® rats treated with urethane (25, 100 and 250mg/kg/day) or saline by oral gavage for 29 days. Additional endpoints integrated into this study were: micronucleated reticulocytes (MN-RET) in peripheral blood; chromosome aberrations (CAb) and DNA damage (%tail intensity via the comet assay) in peripheral blood lymphocytes (PBL); micronucleated polychromatic erythrocytes (MN-PCE) in bone marrow; and DNA damage (comet) in various organs at termination (the 29th dose was added for the comet endpoint at sacrifice). Ethyl methanesulfonate (EMS; 200mg/kg/day on Days 3, 4, 13, 14, 15, 27, 28 and 29) was evaluated as the concurrent positive control (PC). All animals survived to termination and none exhibited overt toxicity, but there were significant differences in body weight and body weight gain in the 250-mg/kg/day urethane group, as compared with the saline control animals. Statistically significant, dose-dependent increases were observed for urethane for: RET CD59– and RBC CD59– (on Days 15 and 29); MN-RET (on Days 4, 15 and 29); and MN-PCE (on Day 29). The comet assay yielded positive results in PBL (Day 15) and liver (Day 29), but negative results for PBL (Days 4 and 29) and brain, kidney and lung (Day 29). No significant increases in PBL CAb were observed at any sample time. Except for PBL CAb (likely due to excessive cytotoxicity), EMS-induced significant increases in all endpoints/tissues. These results compare favorably with earlier in vivo observations and demonstrate the utility and sensitivity of the Pig-a in vivo gene mutation assay, and its ability to be easily integrated, along with other standard genotoxicity endpoints, into 28-day rodent toxicity studies.

Description: The Pig-a assay has shown promise as a regulatory assay for evaluating in vivo gene mutation. A recent International Workshop on Genotoxicity Testing workgroup discussed the state of the assay and identified several knowledge gaps in assay development. This Mutagenesis Special Topic includes a collection of reports that addresses some of these knowledge gaps, including identifying the mutations responsible for the Pig-a mutant phenotype, the effect of sex on the response, probing the robustness of the assay and expanding the number of agents tested in the assay, especially agents expected to yield negative responses.

Description: The Pig-a assay has rapidly gained international interest as a useful tool for assessing the mutagenic potential of compounds in vivo . Although a large number of compounds, including both mutagens and non-mutagens, have been tested in the rat Pig-a assay in haematopoietic cells, there is limited understanding of how perturbations in haematopoiesis affect assay performance. Of particular concern is the possibility that regenerative haematopoiesis alone, without exposure to a genotoxic agent, could result in elevated Pig-a mutant cell frequencies. To address this concern, Wistar-Han rats were dosed by oral gavage with a non-genotoxic haemolytic agent, 2-butoxyethanol (2-BE). Dose levels ranging from 0 to 450mg/kg were tested using both single administration and 28-day treatment regimens. Haematology parameters were assessed at minimum within the first 24h of treatment and 8 days after the final administration. Pig-a mutant frequencies were assessed on Days 15 and ~30 for both treatment protocols and also on Days 43 and 57 for the 28-day protocol. Even at doses of 2-BE that induced marked intravascular lysis and strong compensatory erythropoiesis, the average Pig-a mutant phenotype red blood cell and reticulocyte frequencies were within the historical vehicle control distribution. 2-BE therefore showed no evidence of in vivo mutagenicity in these studies. The data suggest that perturbations in haematopoiesis alone do not lead to an observation of increased mutant frequency in the Pig-a assay.

Description: The Pig-a assay is used for monitoring somatic cell mutation in laboratory animals and humans. The assay detects haematopoietic cells deficient in glycosylphosphatidylinositol (GPI)-anchored protein surface markers using flow cytometry. However, given that synthesis of the protein markers (and the expression of their genes) is independent of the expression of the X-linked Pig-a gene and the function of its enzyme product, the deficiency of markers at the surface of the cells may be caused by a number of events (e.g. by mutation or epigenetic silencing in the marker gene itself or in any of about two dozen autosomal genes involved in the synthesis of GPI). Here we provide direct evidence that the deficiency of the GPI-anchored surface marker CD48 in rat T-cells is accompanied by mutation in the endogenous X-linked Pig-a gene. We treated male F344 rats with N -ethyl- N -nitrosourea (ENU), and established colonies from flow cytometry-identified and sorted CD48-deficient spleen T-lymphocytes. Molecular analysis confirmed that the expanded sorted cells have mutations in the Pig-a gene. The spectrum of Pig-a mutation in our model was consistent with the spectrum of ENU-induced mutation determined in other in vivo models, mostly base-pair substitutions at A:T with the mutated T on the non-transcribed strand of Pig-a genomic DNA. We also used next generation sequencing to derive a similar mutational spectrum from a pool of 64 clones developed from flow-sorted CD48-deficient lymphocytes. Our findings confirm that Pig-a assays detect what they are designed to detect—gene mutation in the Pig-a gene.

Description: Validation of the Pig-a gene mutation assay has been based mainly on studies in male rodents. To determine if the mutagen-induced responses of the X-linked Pig-a gene differ in females compared to males, 7- or 14-week old male and female Sprague Dawley rats were exposed to N -ethyl- N -nitrosourea (ENU). In the study with the 7-week old rats, exposure was to 0, 1, 5 or 25mg ENU/kg/day for three consecutive days (study Days 1–3). Pig-a mutant phenotype reticulocyte (RET CD59– ) and mutant phenotype erythrocyte (RBC CD59– ) frequencies were determined on study Days –4, 15, 29 and 46 using immunomagnetic separation in conjunction with flow cytometric analysis ( In Vivo MutaFlow ® ). Additionally, blood samples collected on Day 4 were analysed for micronucleated reticulocyte (MN-RET) frequency ( In Vivo MicroFlow ® ). The percentage of reticulocytes (%RET) was markedly higher in the 7-week old males compared to females through Day 15 (2.39-fold higher on Day –4). At 25mg/kg/day, ENU reduced Day 4 RET frequencies in both sexes, and the two highest dose levels resulted in elevated MN-RET frequencies, with no sex or treatment x sex interaction. The two highest dose levels significantly elevated the frequencies of mean RET CD59– and RBC CD59– in both sexes from Day 15 onward. RET CD59– and RBC CD59– frequencies were somewhat lower for females compared to males at the highest dose level studied, and differences in RET CD59– resulted in a statistically significant interaction effect of treatment x sex. In the study with 14-week old rats, treatment was for 3 days with 0 or 25mg ENU/kg/day. RET frequencies differed to a lesser degree between the sexes, and in this case there was no evidence of a treatment x sex interaction. These results suggest that the slightly higher response in younger males than in the younger females may be related to differences in erythropoiesis function at that age. In conclusion, while some quantitative differences were noted, there were no qualitative differences in how males and females responded to a prototypical mutagen, and support the contention that both sexes are equally acceptable for Pig-a gene mutation studies.

Description: Isothiocyanates are plant-derived compounds that may be beneficial in the prevention of certain chronic diseases. Yet, by stimulating the production of reactive oxygen species (ROS), isothiocyanates can be genotoxic. Whether antioxidants influence isothiocyanate-induced genotoxicity is unclear, but this situation was clarified appreciably herein. In HCT116 cells, phenethyl isothiocyanate (PEITC) increased ROS production, which was inhibited by N -acetylcysteine (NAC) and deferoxamine (DFO) but not by ascorbic acid (ASC) and trolox (TRX) that were found to be more potent radical scavengers. Surprisingly, ASC and TRX each intensified the DNA damage that was caused by PEITC, but neither ASC nor TRX by themselves caused any DNA damage. In contrast, NAC and DFO each not only attenuated PEITC-induced DNA damage but also attenuated the antioxidant-intensified, PEITC-induced DNA damage. To determine if the DNA damage could be related to possible changes in the major antioxidant defence system, glutathione (GSH) was investigated. PEITC lowered GSH levels, which was prevented by NAC, whereas ASC, TRX and DFO neither inhibited nor enhanced the GSH-lowering effect of PEITC. The GSH synthesis inhibitor, buthionine sulphoxime, intensified PEITC-induced DNA damage, although by itself buthionine sulphoxime did not directly cause DNA damage. The principal findings suggest that ASC and TRX make PEITC more genotoxic, which might be exploited in killing cancer cells as one approach in killing cancer cells is to extensively damage their DNA so as to initiate apoptosis.

Description: The mycotoxin aflatoxin B 1 (AFB 1 ) may initiate cancer by causing oxidatively damaged DNA, specifically by causing 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-oxodG) lesions. Base excision repair removes these lesions, with 8-oxoguanine glycosylase (OGG1) being the rate-limiting enzyme. The aim of this study was to determine the effect of ogg1 deficiency on AFB 1 -induced oxidatively damaged DNA and tumourigenesis. Female wild-type, heterozygous and homozygous ogg1 null mice were given a single dose of 50mg/kg AFB 1 or 40 µl dimethyl sulfoxide (DMSO) ip. Neither ogg1 genotype nor AFB 1 treatment affected levels of oxidised guanine in lung or liver 2h post-treatment. AFB 1 -treated ogg1 null mice showed exacerbated weight loss and mortality relative to DMSO-treated ogg1 null mice, but AFB 1 treatment did not significantly increase lung or liver tumour incidence compared with controls, regardless of ogg1 genotype. Suspect lung masses from three of the AFB 1 -treated mice were adenomas, and masses from two of the mice were osteosarcomas. No osteosarcomas were observed in DMSO-treated mice. All liver masses from AFB 1 -treated mice were adenomas, and one also contained a hepatocellular carcinoma. In DNA from the lung tumours, the K- ras mutation pattern was inconsistent with initiation by AFB 1 . In conclusion, ogg1 status did not have a significant effect on AFB 1 -induced oxidatively damaged DNA or tumourigenesis, but deletion of one or both alleles of ogg1 did increase susceptibility to other aspects of AFB 1 toxicity.

Description: Integration of in vivo genotoxicity testing into standard toxicology studies presents multiple advantages as it reduces animal use and costs, accelerates data generation and provides concurrent data that are useful for interpreting results. The in vivo Pig-a assay is a mammalian gene mutation assay that utilises peripheral blood and thus has a high integration potential. Although inter-laboratory reproducibility has been well demonstrated, further characterisation is required for this assay. In this study, we evaluated intra-laboratory reproducibility of the in vivo Pig-a gene mutation assay (MutaFlow® kit) in rats through the conduct of an assay characterisation prior to subsequent use in Good Laboratory Practices (GLP) toxicology studies. To evaluate intra-laboratory reproducibility, intra-assay and inter-assay variability, ruggedness, robustness and blood storage stability were assessed. These assessments were performed using blood obtained from male Sprague–Dawley rats exposed to 0, 20 or 40mg/kg/day N -ethyl- N -nitrosourea via oral gavage for three consecutive days. Blood was collected from these rats on multiple occasions from Day 29 to Day 71 and samples were analysed for Pig-a mutation using the rat MutaFlow kit. Frequencies of reticulocytes (RET), mutant phenotype erythrocytes (RBC CD59– ) and mutant phenotype RET (RET CD59– ) were determined. Overall, the proportion of RET and frequencies of RBC CD59– and of RET CD59– were consistent throughout the different assessments. The assay demonstrated acceptable intra-run and inter-run variability with coefficients of variation of ≤4.8 and 20.6%, respectively. The method was shown to be independent of the analyst performing the assay and unaffected by small changes in assay conditions. Comparable results were obtained from freshly collected samples and samples refrigerated for up to 4 days. Although technically challenging, the rat Pig-a gene mutation assay using a high-throughput automated method was shown to be reliable. The different assay parameters evaluated during the conduct of this study yielded acceptable results. Thus, the method was considered suitable for use in GLP toxicology studies.

Description: Research over the years has generated enough evidence to implicate areca nut, as a carcinogen in humans. Besides oral, significant rise in the incidence of cancers of the oesophagus, liver and stomach was seen among areca nut chewers. Early diagnosis seems key to understand the initial processes of carcinogenesis which is highly curable. In North-East India, betel quid contains raw areca nut (RAN), lime and small portion of betel leaf without any other constituents. This study was not intended to isolate any active ingredients from the RAN and to look its action. The present objective is to validate the screening of precocious anaphase and analysis of expression of Securin and p53 in non-target cells like human peripheral blood lymphocytes (PBLs) and mouse bone marrow cells (BMCs) as early indicative parameters of RAN + lime-induced cancers. A total of 35 mice were examined at different time points for following ad libitum administration of RAN extract in drinking water with lime. Peripheral blood was collected from 32 human donors of which, 24 were RAN + lime heavy chewers. Expression of genes was assessed by immunoblotting and/or by immunohistochemistry. Histological preparation of stomach tissue of mice revealed that RAN + lime induced stomach cancer. A gradual increase in the frequency of precocious anaphases and aneuploid cells was observed in both RAN + lime-treated mouse BMC and human PBL of RAN heavy chewers. Levels of p53 and Securin were increased in these cells during early days of RAN + lime exposure. The level of Securin was significantly higher in human tumour samples than their adjacent normal counterpart. The expression of Securin was increased significantly in RAN + lime-administered mice as well as in stomach tumour. Present study revealed that precocious anaphase and expression of p53 and Securin in non-target cells are significantly associated with an increased risk of RAN-induced cancer and thus these parameters can be of early diagnostic value.

Description: Silver nanoparticles (AgNPs) are one of the most widely used nanomaterials due to their antibacterial properties. Owing to the recent boost in the usage of AgNPs-containing products, human exposure to AgNPs is increasing, highlighting the need for careful evaluation of AgNPs toxicity in humans. We used two cellular models, hepatic HepG2 and epithelial A549 cell lines, to study the mechanism of AgNPs-induced toxicity at the cellular level. These two cell lines differ significantly in their response to AgNPs treatment. In the case of A549 cells, a minor decrease in viability and increase in the extent of DNA breakage were observed. A markedly different response to AgNPs was observed in HepG2 cells. In short term, a massive induction of DNA breakage was observed, suggesting that the basal activity of antioxidant defence in these cells was not sufficient to effectively protect them from the nanoparticle-induced oxidative stress. After prolonged exposure, the extent of DNA breakage decreased to the level observed in the control cells proving that a successful adaptation to the new conditions had taken place. The cells that were unable to adapt must have died, as revealed by the Neutral Red assay that indicated less than half viable cells after 24-h treatment with 100 µg/ml of 20nm AgNPs. The gene expression analysis revealed that the observed adaptation was underlain by a pro-proliferative, anti-apoptotic signal leading to up-regulation of the genes promoting proliferation and inflammatory response ( EGR1 , FOS , JUN , HK2 , IL4 , MMP10 , VEGFA , WISP1 , CEBPB , IL8 , SELPLG ), genes coding the anti-apoptotic proteins ( BCL2A1 , CCL2 ) and factors involved in the response to stress ( HSPB1 , GADD45A ). Such a selection of highly resistant population of cells should be taken into account in the case of medical applications of nanoparticles since the sustained proliferative signalling and resistance to cell death are hallmarks of cancer, acquired by the cells in the process of carcinogenesis.

Description: Ionising radiation induces single-strand breaks, double-strand breaks (DSB) and base damages in human cell. DSBs are the most deleterious and if not repaired may lead to genomic instability and cell death. DSB can be repaired through non-homologous end joining (NHEJ) pathway in resting lymphocytes. In this study, NHEJ genes and proteins were studied in irradiated human peripheral blood mononuclear cells (PBMC) at resting stage. Dose-response, time point kinetics and adaptive-response studies were conducted in irradiated PBMC at various end points such as DNA damage quantitation, transcription and protein expression profile. Venous blood samples were collected from 20 random, normal and healthy donors with written informed consent. PBMC was separated and irradiated with various doses between 0.1 and 2.0 Gy ( 60 CO- source) for dose-response study. Repair kinetics of DNA damage and time point changes in expression of genes and proteins were studied in post-irradiated PBMC at 2.0 Gy at various time points up to 240min. Adaptive-response study was conducted with a priming dose of 0.1 Gy followed by a challenging dose of 2.0 Gy after 4-h incubation. Our results revealed that Ku70, Ku80, XLF and Ligase IV were significantly upregulated ( P 〈 0.05) at 4-h post-irradiation at transcript and protein level. Adaptive-response study showed significantly increased expression of the proteins involved in NHEJ, suggesting their role in adaptive response in human PBMC at G 0 /G 1, which has important implications to human health.

Description: Excision repair cross complementing group 1 (ERCC1) and X-ray repair cross-complementing groups 1 (XRCC1) are DNA repair enzymes. Polymorphisms in DNA repair genes may be important factors affecting cancer susceptibility, prognosis and therapy outcome. The purpose of this study was to investigate the correlation of ERCC1 and XRCC1 polymorphisms with colorectal cancer (CRC) risk, and explore the effect of polymorphisms on event-free, overall survival and oxaliplatin-based therapy in CRC patients. Genotyping was examined with the iMLDR technique. An unconditional logistic regression model was used to estimate the association of certain polymorphisms with CRC risk. The Kaplan–Meier method, log-rank test and Cox regression model were employed to evaluate the effects of polymorphisms on survival analysis. Results showed that Trp/Trp genotype of XRCC1 Arg194Trp and AA genotype of ERCC1 rs2336219 have a significantly increased risk of CRC; Trp allele of XRCC1 Arg194Trp and CC genotype of ERCC1 rs735482 were associated with lower response to oxaliplatin-based chemotherapy, a shorter survival and a higher risk of relapse or metastasis. 194Trp/280Arg/399Arg haplotype was associated with a significant resistance, and the ERCC1 protein expression was statistically higher in tumours with rs735482 CC genotype than with AA genotype. Our studies indicate that XRCC1 and ERCC1 polymorphisms probably affect susceptibility, chemotherapy response and survival of CRC patients.

Description: Epigenetic control of gene expression in children remains poorly understood, but new technologies can help elucidate the relationship between expression and DNA methylation. Here, we utilized the nCounter Analysis System to characterise the expression of 60 genes in 69 9-year-old children from a cohort with a high prevalence of obesity. nCounter expression levels ranged broadly (from 3 to over 10000 messenger RNA counts) and were divided into four categories: high (〉2000 counts), moderate (200–1000 counts), low (100–200 counts) and marginal (〈100 counts). For a subset of five genes ( ADIPOR1 , PPARG1 , GSTM1 , PON1 and ACACA ) from different expression level categories, we validated nCounter data using reverse transcription-polymerase chain reaction (RT-PCR), and expanded RT-PCR analysis of ADIPOR1 to include 180 children. Expression data from the two methodologies were correlated for all five genes included in the validation experiment, with estimates ranging from r s = 0.26 ( P = 0.02) to r s = 0.88 ( P 〈 5 x 10 –6 ). ADIPOR1 and PPARG1 nCounter expression levels were negatively correlated ( r = –0.60, P 〈 5 x 10 –5 ), and this relationship was stronger in overweight children ( r = –0.73, P 〈 5 x 10 –5 ) than in normal weight children ( r = –0.42, P = 0.016). Using methylation data from the Infinium HumanMethylation450 BeadChip ( n = 180), we found eight CpG sites in ADIPOR1 and PPARG where methylation level was associated with expression by RT-PCR ( P 〈 0.05). Hypomethylation of PPARG gene body site cg10499651 was associated with increased expression as measured by both RT-PCR and nCounter ( P 〈 0.05). We found no statistically significant relationships between either expression or methylation of ADIPOR1 and PPARG and body mass index or waist circumference. In addition to demonstrating the validity of expression data derived from nCounter, our results illustrate the use of new technologies in assessing epigenetic effects on expression in children.

Description: The in vitro micronucleus test is a well-known test for the screening of genotoxic compounds. However until now, most studies have been performed on either human peripheral lymphocytes or established cancer cell lines. This study provides human mesenchymal stem cells as an alternative to the conventional micronucleus test. We grew umbilical cord mesenchymal stem cells (UC-MSCs) on coverslips eliminating the cumbersome technique involving hypotonic treatment, fixation and preparing smears required for suspension culture (lymphocytes). The background frequency of nuclear blebs and micronuclei in UC-MSCs was found to be 7±5, in lymphocytes 16±3.5 and 9±3 and that for A549 cell line was 65±5 and 15±5 per 1000 cells, respectively, suggesting differences in the repair mechanism of normal and cancer cell lines. We inspected the cytotoxic and genotoxic effects of two known mutagens, mitomycin-C and hydrogen peroxide (H 2 O 2 ), on UC-MSCs, lymphocytes and A549 cells. Treatment with mitomycin-C and H 2 O 2 demonstrated drastic differences in the degree of cytotoxicity and genotoxicity suggesting a constitutional difference between normal and cancer cells. In addition we tested two solvents, dimethyl sulfoxide (DMSO) and ethanol, and two drugs, metformin and rapamycin. DMSO above 1% was found to be cytotoxic and genotoxic, whereas ethanol at same concentration was neither cytotoxic nor genotoxic indicating the minimal non-toxic level of the solvents. This study thus offers UC-MSCs as a better substitute to peripheral lymphocytes and cancer cell lines for high throughput screening of compounds and reducing the animal studies.

Description: Cells exhibiting radiation-induced genomic instability exhibit varied spectra of genetic and chromosomal aberrations. Even so, oxidative stress remains a common theme in the initiation and/or perpetuation of this phenomenon. Isolated oxidatively modified bases, abasic sites, DNA single strand breaks and clustered DNA damage are induced in normal mammalian cultured cells and tissues due to endogenous reactive oxygen species generated during n ormal cellular metabolism in an aerobic environment. While sparse DNA damage may be easily repaired, clustered DNA damage may lead to persistent cytotoxic or mutagenic events that can lead to genomic instability. In this study, we tested the hypothesis that DNA damage signatures characterised by altered levels of endogenous, potentially mutagenic, types of DNA damage and chromosomal breakage are related to radiation-induced genomic instability and persistent oxidative stress phenotypes observed in the chromosomally unstable progeny of irradiated cells. The measurement of oxypurine, oxypyrimidine and abasic site endogenous DNA damage showed differences in non-double-strand breaks (DSB) clusters among the three of the four unstable clones evaluated as compared to genomically stable clones and the parental cell line. These three unstable clones also had increased levels of DSB clusters. The results of this study demonstrate that each unstable cell line has a unique spectrum of persistent damage and lead us to speculate that alterations in DNA damage signaling and repair may be related to the perpetuation of genomic instability.

Description: We give a bordism-theoretic characterization of those closed almost contact $(2q{+ }1)$ -manifolds (with $q\geq 2$ ) that admit a Stein fillable contact structure. Our method is to apply Eliashberg's $h$ -principle for Stein manifolds in the setting of Kreck's modified surgery. As an application, we show that any simply connected almost contact 7-manifold with torsion-free second homotopy group is Stein fillable. We also discuss the Stein fillability of exotic spheres and examine subcritical Stein fillability.

Description: Casson-type invariants emerging from Donaldson theory over certain negative-definite four-manifolds were recently suggested by Teleman. These are defined by an algebraic count of points in a zero-dimensional moduli space of flat instantons. Motivated by the cobordism programme of proving Witten's conjecture, we use a moduli space of ${\rm PU}(2)$ Seiberg–Witten monopoles to exhibit an oriented one-dimensional cobordism of the instanton moduli space to the empty space. The Casson-type invariant must therefore vanish.

Description: The dynamical and stationary behaviors of a fourth-order equation in the unit ball with clamped boundary conditions and a singular reaction term are investigated. The equation arises in the modeling of microelectromechanical systems and includes a positive voltage parameter $\lambda$ . It is shown that there is a threshold value $\lambda _* 〉 0$ of the voltage parameter such that no radially symmetric stationary solution exists for $\lambda 〉 \lambda _* $ , while at least two such solutions exist for $\lambda \in (0,\lambda _* )$ . Local and global well-posedness results are obtained for the corresponding hyperbolic and parabolic evolution problems as well as the occurrence of finite time singularities when $\lambda 〉 \lambda _* $ .

Description: Assuming the generalized Riemann hypothesis, we prove a quantitative estimate for the number of simple zeros on the critical line for $L$ -functions attached to classical holomorphic newforms.

Description: In this paper, we consider a $\mathbb {Q}$ -Fano $3$ -fold weighted complete intersection of codimension $2$ in the $85$ families listed in Iano-Fletcher's list and determine which cycle is a maximal center or not. For each maximal center, we construct either a birational involution which untwists the maximal singularity or a Sarkisov link centered at the cycle to another explicitly described Mori fiber space. As a consequence, nineteen families are proved to be birationally rigid and the remaining $66$ families are proved to be birationally non-rigid.

Description: Suppose that a sequence of numbers $x_n$ (a ‘signal’) is transmitted through a noisy channel. The receiver observes a noisy version of the signal with additive random fluctuations, $x_n + \xi _n$ , where $\xi _n$ is a sequence of independent standard Gaussian random variables. Suppose further that the signal is known to come from some fixed space ${\mathscr {X}}$ of possible signals. Is it possible to fully recover the transmitted signal from its noisy version? Is it possible to at least detect that a non-zero signal was transmitted? In this paper, we consider the case in which signals are infinite sequences and the recovery or detection are required to hold with probability 1. We provide conditions on the space ${\mathscr {X}}$ for checking whether detection or recovery are possible. We also analyze in detail several examples including spaces of Fourier transforms of measures, spaces with fixed amplitudes and the space of almost periodic functions. Many of our examples exhibit critical phenomena, in which a sharp transition is made from a regime in which recovery is possible to a regime in which even detection is impossible.

Description: In this paper, we study the semi-stable subcategories of the category of representations of a Euclidean quiver, and the possible intersections of these subcategories. Contrary to the Dynkin case, we find out that the intersection of semi-stable subcategories may not be semi-stable. However, only a finite number of exceptions occur, and we give a description of these subcategories. Moreover, one can attach a simplicial fan in $\mathbb {Q}^n$ to any acyclic quiver $Q$ , and this simplicial fan allows one to completely determine the canonical presentation of any element in $\mathbb {Z}^n$ . This fan has a nice description in the Dynkin and Euclidean cases: it is described using an arrangement of convex codimension-1 subsets of $\mathbb {Q}^n$ , each such subset being indexed by a real Schur root or a set of quasi-simple objects. This fan also characterizes when two different stability conditions give rise to the same semi-stable subcategory.

Description: The $j$ -multiplicity plays an important role in the intersection theory of Stückrad–Vogel cycles, while recent developments confirm the connections between the $\epsilon$ -multiplicity and equisingularity theory. In this paper, we establish, under some constraints, a relationship between the $j$ -multiplicity of an ideal and the degree of its fiber cone. As a consequence, we are able to compute the $j$ -multiplicity of all the ideals defining rational normal scrolls. By using the standard monomial theory, we can also compute the $j$ - and $\epsilon$ -multiplicity of ideals defining determinantal varieties: The found quantities are integrals which, quite surprisingly, are central in random matrix theory.

Description: Let $\pi : X \to Y$ be a morphism of projective varieties and suppose that $\alpha $ is a pseudo-effective numerical cycle class satisfying $\pi _{*}\alpha =0$ . A conjecture of Debarre, Jiang, and Voisin predicts that $\alpha $ is a limit of classes of effective cycles contracted by $\pi $ . We establish new cases of the conjecture for higher codimension cycles. In particular, we prove a strong version when $X$ is a fourfold and $\pi $ has relative dimension 1.

Description: Let $R$ be a group of prime order $r$ that acts on the $r'$ -group $G$ , let $RG$ be the semidirect product of $G$ with $R$ , let ${\mathbb {F}}$ be a field and $V$ be a faithful completely reducible $\mathbb {F}[{RG}]$ -module. Trivially, $C_{G}({R})$ acts on $C_{V}({R})$ . Let $K$ be the kernel of this action. What can be said about $K$ ? This question is considered when $G$ is soluble. It turns out that $K$ is subnormal in $G$ or $r$ is a Fermat or half-Fermat prime. In the latter cases, the subnormal closure of $K$ in $G$ is described. Several applications to the theory of automorphisms of soluble groups are given.

Description: The mono-epi (ME) exact structure on the morphisms of an exact category $(\mathcal {A}; \mathcal {E})$ is introduced and used to prove ideal versions of Salce's Lemma, Christensen's (Ghost) Lemma, and Wakamatsu's Lemma for an exact category. Salce's Lemma establishes a bijective correspondence $\mathcal {I} \mapsto \mathcal {I}^{\perp }$ between the class of special precovering ideals of $(\mathcal {A}; \mathcal {E})$ and that of its special preenveloping ideals. ME-extensions of morphisms are used to define an extension $\mathcal {I} \diamond \mathcal {J}$ of ideals. Christensen's Lemma asserts that the class of special precovering (respectively, special preenveloping) ideals is closed under products and extensions and that the bijective correspondence of Salce's Lemma satisfies $(\mathcal {I} \mathcal {J})^{\perp } = \mathcal {J}^{\perp } \diamond \mathcal {I}^{\perp }$ and $(\mathcal {I} \diamond \mathcal {J})^{\perp } = \mathcal {J}^{\perp } \mathcal {I}^{\perp }.$ Wakamatsu's Lemma asserts that if a covering ideal $\mathcal {I}$ is closed under ME-extensions, then it is a special precovering ideal. As an application, it is proved that if $G$ is a finite group and $\Phi $ is the ideal of phantom morphisms in the category $k[G]$ - $\rm Mod,$ then $\Phi ^{n-1}$ is the object ideal generated by projective modules, where $n$ is the nilpotency index of the Jacobson radical $J.$ If $R$ is a semiprimary ring, with $J^n =0,$ then $\Phi ^n$ is generated by projective modules. For a right coherent ring $R$ over which every cotorsion left $R$ -module has a coresolution of length $n$ by pure injective modules, $\Phi ^{n+1}$ is generated by flat modules.

Description: We complete the equisingular deformation classification of irreducible singular plane sextic curves. As a by-product, we also compute the fundamental groups of the complement of all but a few maximizing sextics.

Description: Let $P_{n}(x)= \sum _{i=0}^n \xi _i x^i$ be a Kac random polynomial where the coefficients $\xi _i$ are i.i.d. copies of a given random variable $\xi $ . Our main result is an optimal quantitative bound concerning real roots repulsion. This leads to an optimal bound on the probability that there is a real double root. As an application, we consider the problem of estimating the number of real roots of $P_n$ , which has a long history and in particular was the main subject of a celebrated series of papers by Littlewood and Offord from the 1940s. We show, for a large and natural family of atom variables $\xi $ , that the expected number of real roots of $P_n(x)$ is exactly $({2}/{\pi }) \log n +C +o(1)$ , where $C$ is an absolute constant depending on the atom variable $\xi $ . Prior to this paper, such a result was known only for the case when $\xi $ is Gaussian.

Description: The goal of this article was to study the Iwasawa theory of an abelian variety $A$ that has complex multiplication by a complex multiplication (CM) field $F$ that contains the reflex field of $A$ , which has supersingular reduction at every prime above $p$ . To do so, we make use of the signed Coleman maps constructed in our companion article [Kâzım Büyükboduk and Antonio Lei, ‘Integral Iwasawa theory of motives for non-ordinary primes’, 2014, in preparation, draft available upon request] to introduce signed Selmer groups as well as a signed $p$ -adic $L$ -function via a reciprocity conjecture that we formulate for the (conjectural) Rubin–Stark elements (which is a natural extension of the reciprocity conjecture for elliptic units). We then prove a signed main conjecture relating these two objects. To achieve this, we develop along the way a theory of Coleman-adapted rank- $g$ Euler–Kolyvagin systems to be applied with Rubin–Stark elements and deduce the main conjecture for the maximal $\mathbb {Z}_p$ -power extension of $F$ for the primes failing the ordinary hypothesis of Katz.

Description: We introduce a notion of rough paths on embedded submanifolds and demonstrate that this class of rough paths is natural. On the way, we develop a notion of rough integration and an efficient and intrinsic theory of rough differential equations (RDEs) on manifolds. The theory of RDEs is then used to construct parallel translation along manifold-valued rough paths. Finally, this framework is used to show that there is a one-to-one correspondence between rough paths on a $d$ -dimensional manifold and rough paths on $d$ -dimensional Euclidean space. This last result is a rough path analogue of Cartan's development map and its stochastic version which was developed by Eells and Elworthy and Malliavin.

Description: We show that several important normal subgroups $\Gamma $ of the mapping class group of a surface satisfy the following property: any free, ergodic, probability measure-preserving action $\Gamma \curvearrowright X$ is stably $OE$ -superrigid. These include the central quotients of most surface braid groups and most Torelli groups and Johnson kernels. In addition, we show that all these groups satisfy the measure equivalence rigidity and we describe all their lattice-embeddings. Using these results in combination with previous results from Chifan–Ioana–Kida [‘ $W^*$ -superrigidity for arbitrary actions of central quotients of braid groups’, Math. Ann. 361 (2015) 925–959], we deduce that any free, ergodic, probability measure-preserving action of almost any surface braid group is stably $W^*$ -superrigid, that is, it can be completely reconstructed from its von Neumann algebra.

Description: Motivated by recent work in the mathematics and engineering literature, we study integrability and non-tangential regularity on the two-torus for rational functions that are holomorphic on the bidisk. One way to study such rational functions is to fix the denominator and look at the ideal of polynomials in the numerator such that the rational function is square integrable. A concrete list of generators is given for this ideal as well as a precise count of the dimension of the subspace of numerators with a specified bound on bidegree. The dimension count is accomplished by constructing a natural pair of commuting contractions on a finite-dimensional Hilbert space and studying their joint generalized eigenspaces. Non-tangential regularity of rational functions on the polydisk is also studied. One result states that rational inner functions on the polydisk have non-tangential limits at every point of the $n$ -torus. An algebraic characterization of higher non-tangential regularity is given. We also make some connections with the earlier material and prove that rational functions on the bidisk which are square integrable on the two-torus are non-tangentially bounded at every point. Several examples are provided.

Description: In a previous study, we validated an in vitro genotoxicity assay based on H2AX quantification using the In-Cell Western (ICW) method in HepG2 cells. The assay demonstrated high sensitivity and specificity but failed to detect genotoxicity for few compounds that require specific metabolic bioactivation not sufficiently covered by HepG2 cells. The aim of the present study was to assess H2AX ICW sensitivity using a broader range of genotoxic molecules with HepG2 cells and three additional human cell lines with distinct biotransformation properties: two cell lines expressing some phase I and II bioactivation capabilities (LS-174T and Hep3B), and one with poor general bioactivation properties (ACHN). We evaluated the four cell lines by testing 24 compounds recommended by European Centre for the Validation of Alternative Methods and a set of 24 additional chemicals with different mode of genotoxic action (MOA) (aneugenicity, DNA adducts formation, induction of oxidative stress), including some known to require specific cytochrome P450 metabolic bioactivation. Results for the 48 compounds tested showed that the H2AX ICW assay was more sensitive with LS-174T and HepG2 cells than with Hep3B or ACHN cell lines. Among the 38 compounds tested with positive or equivocal carcinogenicity data, 36 (95%) showed a positive genotoxic response with the H2AX ICW assay compared to only 27 (71%) using the Ames assay. We confirm that the H2AX ICW assay on HepG2 cells, without an exogenous metabolic activation system, may be a suitable test to predict the in vivo genotoxicity of chemicals with different genotoxic MOA. Moreover, the use of the ACHN cell line in combination with LS-174T and HepG2 cells may permit in many cases to discriminate direct from bioactivated genotoxins. Overall, our results confirm the high sensitivity of the H2AX ICW assay which, in turn, should reduce the number of animals used for genotoxicity assessment.

Description: Quantum dots (QD) have unique electronic and optical properties promoting biotechnological advances. However, our understanding of the toxicological structure–activity relationships remains limited. This study aimed to determine the biological impact of varying nanomaterial surface chemistry by assessing the interaction of QD with either a negative (carboxyl), neutral (hexadecylamine; HDA) or positive (amine) polymer coating with human lymphoblastoid TK6 cells. Following QD physico-chemical characterisation, cellular uptake was quantified by optical and electron microscopy. Cytotoxicity was evaluated and genotoxicity was characterised using the micronucleus assay (gross chromosomal damage) and the HPRT forward mutation assay (point mutagenicity). Cellular damage mechanisms were also explored, focusing on oxidative stress and mitochondrial damage. Cell uptake, cytotoxicity and genotoxicity were found to be dependent on QD surface chemistry. Carboxyl-QD demonstrated the smallest agglomerate size and greatest cellular uptake, which correlated with a dose dependent increase in cytotoxicity and genotoxicity. Amine-QD induced minimal cellular damage, while HDA-QD promoted substantial induction of cell death and genotoxicity. However, HDA-QD were not internalised by the cells and the damage they caused was most likely due to free cadmium release caused by QD dissolution. Oxidative stress and induced mitochondrial reactive oxygen species were only partially associated with cytotoxicity and genotoxicity induced by the QD, hence were not the only mechanisms of importance. Colloidal stability, nanoparticle (NP) surface chemistry, cellular uptake levels and the intrinsic characteristics of the NPs are therefore critical parameters impacting genotoxicity induced by QD.

Description: We previously reported that a urate-null strain of Drosophila is hypersensitive to cigarette smoke (CS), and we suggested that CS induces oxidative stress in Drosophila because uric acid is a potent antioxidant. Although the carcinogenic risk of CS exposure is widely recognized; documentation of in vivo genotoxic activity of environmental CS, especially gaseous-phase CS, remains inconclusive. To date, somatic-cell mutations in Drosophila resulting from exposure to CS have not been detected via the somatic mutation and recombination test (wing spot test) with wild-type flies, a widely used Drosophila assay for the detection of somatic-cell mutation; moreover, genotoxicity has not been documented via a DNA repair test that involves DNA repair-deficient Drosophila. In this study, we used a new Drosophila strain ( y v ma-l; mwh ) to examine the mutagenicity induced by gaseous-phase CS; these flies are urate-null due to a mutation in ma-l , and they are heterozygous for multiple wing hair ( mwh ), a mutation that functions as a marker for somatic-cell mutation. In an assay with this newly developed strain, a superoxide anion-producing weed-killer, paraquat, exhibited significant mutagenicity; in contrast, paraquat was hardly mutagenic with a wild-type strain. Drosophila larvae were exposed to CS for 2, 4 or 6h, and then kept at 25 ° C on instant medium until adulthood. After eclosion, mutant spots, which consisted of mutant hairs on wings, were scored. The number of mutant spots increased significantly in an exposure time-dependent manner in the urate-null females ( ma-l (–/–)), but not in the urate-positive females ( ma-l (+/–)). In this study, we showed that short-term exposure to CS was mutagenic in this in vivo system. In addition, we obtained suggestive data regarding reactive oxygen species production in larva after CS exposure using the fluorescence probe H 2 DCFDA. These results suggest that oxidative damage, which might be countered by uric acid, was partly responsible for induction of somatic cell mutations in Drosophila larvae exposed to CS.

Description: DNA repair pathways play a critical role in maintaining cellular homeostasis by repairing DNA damage induced by endogenous processes and xenobiotics, including environmental chemicals. Induction of DNA damage may lead to genomic instability, disruption of cellular homeostasis and potentially tumours. Isogenic chicken DT40 B-lymphocyte cell lines deficient in DNA repair pathways can be used to identify genotoxic compounds and aid in characterising the nature of the induced DNA damage. As part of the US Tox21 program, we previously optimised several different DT40 isogenic clones on a high-throughput screening platform and confirmed the utility of this approach for detecting genotoxicants by measuring differential cytotoxicity in wild-type and DNA repair-deficient clones following chemical exposure. In the study reported here, we screened the Tox21 10K compound library against two isogenic DNA repair-deficient DT40 cell lines ( KU70 –/– / RAD54 –/– and REV3 –/– ) and the wild-type cell line using a cell viability assay that measures intracellular adenosine triphosphate levels. KU70 and RAD54 are genes associated with DNA double-strand break repair processes, and REV3 is associated with translesion DNA synthesis pathways. Active compounds identified in the primary screening included many well-known genotoxicants (e.g. adriamycin, melphalan) and several compounds previously untested for genotoxicity. A subset of compounds was further evaluated by assessing their ability to induce micronuclei and phosphorylated H2AX. Using this comprehensive approach, three compounds with previously undefined genotoxicity—2-oxiranemethanamine, AD-67 and tetraphenylolethane glycidyl ether—were identified as genotoxic. These results demonstrate the utility of this approach for identifying and prioritising compounds that may damage DNA.

Description: Cleidocranial dysplasia (CCD; MIM 119600) is an autosomal dominant skeletal dysplasia characterised by hypopalstic and/or aplastic clavicles, midface hypoplasia, absent or delayed closure of cranial sutures, moderately short stature, delayed eruption of permanent dentition and supernumerary teeth. The molecular pathogenesis can be explained in about two-thirds of CCD patients by haploinsufficiency of the RUNX2 gene. In our current study, we identified a novel and rare variant of the RUNX2 gene (c.181_189dupGCGGCGGCT) in a Japanese patient with phenotypic features of CCD. The insertion led an alanine tripeptide expansion (+3Ala) in the polyalanine tract. To date, a RUNX2 variant with alanine decapeptide expansion (+10Ala) is the only example of a causative variant of RUNX2 with polyalanine tract expansion to be reported, whilst RUNX2 (+1Ala) has been isolated from the healthy population. Thus, precise analyses of the RUNX2 (+3Ala) variant were needed to clarify whether the tripeptide expanded RUNX2 is a second disease-causing mutant with alanine tract expansion. We therefore investigated the biochemical properties of the mutant RUNX2 (+3Ala), which contains 20 alanine residues in the polyalanine tract. When transfected in COS7 cells, RUNX2 (+3Ala) formed intracellular ubiquitinated aggregates after 24h, and exerted a dominant negative effect in vitro . At 24h after gene transfection, whereas slight reduction was observed in RUNX2 (+10Ala), all of these mutants significantly activated osteoblast-specific element-2, a cis-acting sequence in the promoter of the RUNX2 target gene osteocalcin. The aggregation growth of RUNX2 (+3Ala) was clearly lower and slower than that of RUNX2 (+10Ala). Furthermore, we investigated several other RUNX2 variants with various alanine tract lengths, and found that the threshold for aggregation may be RUNX2 (+3Ala). We conclude that RUNX2 (+3Ala) is the cause of CCD in our current case, and that the accumulation of intracellular aggregates in vitro is related to the length of the alanine tract.

Description: In Saccharomyces cerevisiae , disruption of genes by deletion allowed elucidation of the molecular mechanisms of a series of human diseases, such as in Wilson disease (WD). WD is a disorder of copper metabolism, due to inherited mutations in human copper-transporting ATPase ( ATP7B ). An orthologous gene is present in S. cerevisiae , CCC2 gene. Copper is required as a cofactor for a number of enzymes. In excess, however, it is toxic, potentially carcinogenic, leading to many pathological conditions via oxidatively generated DNA damage. Deficiency in ATP7B (human) or Ccc2 (yeast) causes accumulation of intracellular copper, favouring the generation of reactive oxygen species. Thus, it becomes important to study the relative importance of proteins involved in the repair of these lesions, such as Ogg1 . Herein, we addressed the influence Ogg1 repair in a ccc2 deficient strain of S. cerevisiae . We constructed ccc 2-disrupted strains from S. cerevisiae ( ogg 1 ccc2 and ccc 2), which were analysed in terms of viability and spontaneous mutator phenotype. We also investigated the impact of 4-nitroquinoline-1-oxide (4-NQO) on nuclear DNA damage and on the stability of mitochondrial DNA. The results indicated a synergistic effect on spontaneous mutagenesis upon OGG1 and CCC2 double inactivation, placing 8-oxoguanine as a strong lesion-candidate at the origin of spontaneous mutations. The ccc2 mutant was more sensitive to cell killing and to mutagenesis upon 4-NQO challenge than the other studied strains. However, Ogg1 repair of exogenous-induced DNA damage revealed to be toxic and mutagenic to ccc2 deficient cells, which can be due to a detrimental action of Ogg1 on DNA lesions induced in ccc2 cells. Altogether, our results point to a critical and ambivalent role of BER mediated by Ogg1 in the maintenance of genomic stability in eukaryotes deficient in CCC2 gene.

Description: We propose a construction of a tensor exact category $\mathcal {F}_X^m$ of Artin–Tate motivic sheaves with finite coefficients $\mathbb {Z}/m$ over an algebraic variety $X$ (over a field $K$ of characteristic prime to $m$ ) in terms of étale sheaves of $\mathbb {Z}/m$ -modules over $X$ . Among the objects of $\mathcal {F}_X^m$ , in addition to the Tate motives $\mathbb {Z}/m(j)$ , there are the cohomological relative motives with compact support $\mathcal {M}_{cc}^m(Y/X)$ of varieties $Y$ quasi-finite over $X$ . Exact functors of inverse image with respect to morphisms of algebraic varieties and direct image with compact supports with respect to quasi-finite morphisms of varieties $Y\longrightarrow X$ act on the exact categories $\mathcal {F}_X^m$ . Assuming the existence of triangulated categories of motivic sheaves $\mathcal {D}\mathcal {M}(X,\mathbb {Z}/m)$ over algebraic varieties $X$ over $K$ and a weak version of the ‘six operations’ in these categories, we identify $\mathcal {F}_X^m$ with the exact subcategory in $\mathcal {D}\mathcal {M}(X,\mathbb {Z}/m)$ consisting of all the iterated extensions of the Tate twists $\mathcal {M}_{cc}^m(Y/X)(j)$ of the motives $\mathcal {M}_{cc}^m(Y/X)$ . An isomorphism of the $\mathbb {Z}/m$ -modules ${\rm Ext}$ between the Tate motives $\mathbb {Z}/m(j)$ in the exact category $\mathcal {F}_X^m$ with the motivic cohomology modules predicted by the Beilinson–Lichtenbaum étale descent conjecture (recently proved by Voevodsky, Rost et al. ) holds for smooth varieties $X$ over $K$ if and only if the similar isomorphism holds for Artin–Tate motives over fields containing $K$ . When $K$ contains a primitive $m$ -root of unity, the latter condition is equivalent to a certain Koszulity hypothesis, as shown in our previous paper [Positselski, ‘Mixed Artin–Tate motives with finite coefficients’, Mosc. Math. J. 11 (2011) 317–402].

Description: We present new algorithms for computing zeta functions of algebraic varieties over finite fields. In particular, let $X$ be an arithmetic scheme (scheme of finite type over $\textbf {Z}$ ), and for a prime $p$ let $\zeta _{X_p}(s)$ be the local factor of its zeta function. We present an algorithm that computes $\zeta _{X_p}(s)$ for a single prime $p$ in time $p^{1/2+o(1)}$ , and another algorithm that computes $\zeta _{X_p}(s)$ for all primes $p 〈 N$ in time $N \log ^{3+o(1)} N$ . These generalise previous results of the author from hyperelliptic curves to completely arbitrary varieties.

Description: Formaldehyde (FA) is a commonly used chemical in anatomy and pathology laboratories as a tissue preservative and fixative. Because of its sensitising properties, irritating effects and cancer implication, FA accounts probably for the most important chemical-exposure hazard concerning this professional group. Evidence for genotoxic effects and carcinogenic properties in humans is insufficient and conflicting, particularly in regard to the ability of inhaled FA to induce toxicity on other cells besides first contact tissues, such as buccal and nasal cells. To evaluate the effects of exposure to FA in human peripheral blood lymphocytes, a group of 84 anatomy pathology laboratory workers exposed occupationally to FA and 87 control subjects were tested for chromosomal aberrations (CAs) and DNA damage (comet assay). The level of exposure to FA in the workplace air was evaluated. The association between genotoxicity biomarkers and polymorphic genes of xenobiotic-metabolising and DNA repair enzymes were also assessed. The estimated mean level of FA exposure was 0.38±0.03 ppm. All cytogenetic endpoints assessed by CAs test and comet assay % tail DNA (%TDNA) were significantly higher in FA-exposed workers compared with controls. Regarding the effect of susceptibility biomarkers, results suggest that polymorphisms in CYP2E1 and GSTP1 metabolic genes, as well as, XRCC1 and PARP1 polymorphic genes involved in DNA repair pathways are associated with higher genetic damage in FA-exposed subjects. Data obtained in this study show a potential health risk situation of anatomy pathology laboratory workers exposed to FA (0.38 ppm). Implementation of security and hygiene measures may be crucial to decrease risk. The obtained information may also provide new important data to be used by health care programs and by governmental agencies responsible for occupational health and safety.

Description: Nitrous oxide (N 2 O) has been widely used as a dental and surgical anaesthetic for over 150 years. However, results from a recent study suggested that increased DNA damage was seen in lymphocytes from surgical patients and this led to its continued clinical use to be questioned. The data can be challenged on technical grounds and must be considered with other studies in order to assess any possible risk. There are other studies indicating that N 2 O has weak genotoxicity in man, but these are confused by exposure of the populations to other anaesthetic gases including isoflurane and sevoflurane, both of which have also been reported to increase DNA damage. It should be noted that the suggested genotoxic mechanisms are all indirect, including folate deficiency, oxidative stress and homocysteine toxicity. Further, results from in vitro studies indicate that N 2 O has no direct DNA reactivity as negative results were obtained in a bacterial mutation (Ames) test and an assay for mutation at the hprt locus in Chinese hamster lung cells. Although not performed to definitive study designs, no evidence of carcinogenicity was seen in two long-term tests in mice and another in rats. Although there is some evidence that N 2 O is weakly genotoxic in humans, this appears to be similar to that reported for isoflurane and sevoflurane and all the postulated mechanisms have clear thresholds with no evidence of direct DNA reactivity. Because any potential genotoxic mechanism would have a threshold, it seems reasonable to conclude that neither occasional high exposure to patients as an anaesthetic nor low-level exposure to staff within published recommended exposure limits presents any significant carcinogenic risk.

Description: A high risk of neoplastic transformation of nasal and paranasal sinuses mucosa is related to the occupational exposure to wood dust. However, the role of occupational exposures in the aetiology of the airway cancers remains largely unknown. Here, an in vitro model was performed to investigate the carcinogenic effect of wood dusts. Human bronchial epithelial cells were incubated with hard and soft wood dusts and the DNA damage and response to DNA damage evaluated. Wood dust exposure induced accumulation of oxidised DNA bases, which was associated with a delay in DNA repair activity. By exposing cells to wood dust at a prolonged time, wood dust-initiated cells were obtained. Initiated-cells were able to form colonies in soft agar, and to induce blood vessel formation. These cells showed extensive autophagy, reduced DNA repair, which was associated with reduced OGG1 expression and oxidised DNA base accumulation. These events were found related to the activation of EGFR/AKT/mTOR pathway, through phosphorylation and subsequent inactivation of tuberin. The persistence in the tissue of wood dusts, their repetitious binding with EGFR may continually trigger the activation switch, leading to chronic down-regulation of genes involved in DNA repair, leading to cell transformation and proliferation.

Description: We investigated the inflammatory response, acute phase response and genotoxic effect of diesel exhaust particles (DEPs, NIST1650b) following a single intratracheal instillation. C57BL/6J BomTac mice received 18, 54 or 162 µg/mouse and were killed 1, 3 and 28 days post-exposure. Vehicle controls and the benchmark particle carbon black (CB, Printex 90; 162 µg/mouse) were tested alongside for comparison. The cellular composition and protein concentration were determined in bronchoalveolar lavage (BAL) fluid as markers for an inflammatory response. Pulmonary and systemic genotoxicity was analysed by the alkaline comet assay as DNA strand breaks in BAL cells, lung and liver tissue. The pulmonary acute phase response was analysed by Saa3 mRNA levels by real-time quantitative polymerase chain reaction. Instillation of DEP induced a strong neutrophil influx 1 and 3 days, but not 28 days post-exposure. Saa3 mRNA levels were increased at all time point for the highest dose and 28 days post-exposure for the middle dose. DEP increased levels of DNA strand breaks in lung tissue for all doses 1 day post-exposure and after 28 days for mid- and high-dose groups. Pulmonary exposure to DEP induced transient inflammation but long-lasting pulmonary acute phase response as well as genotoxicity in lung tissue 28 days post-exposure. The observed long-term pulmonary genotoxicity by DEP was less than the previously observed genotoxicity for CB using identical experimental set-up.

Description: As a toxic secondary metabolite of Aspergillus species, Aflatoxin B1 (AFB1) is a major food and feed contaminant in tropical and sub-tropical regions with high temperature and humidity. It has been reported to be toxic to the female reproductive system in laboratory and domestic animals. In the present study, the influence of acute exposure to AFB1 (10 and 50 μM, 44h) on porcine oocyte maturation and its possible mechanism were investigated. The maturation rates of oocytes decreased significantly in the presence of 50 μM of AFB1. Cell cycle analysis showed that most oocytes were arrested at germinal vesicle breakdown or meosis I stage. However, actin assembly, spindle structure and chromosome alignment were not disrupted after exposure to 50 μM AFB1. Further study showed that DNA methylation levels increased in treated oocytes (50 μM). Histone methylation levels were also analysed after treatment (50 μM): H3K27me3 and H3K4me2 levels decreased, whereas H3K9me3 level increased, indicating that epigenetic modification was affected. AFB1 treatment (50 μM) also induced oxidative stress and further led to autophagy, as shown by accumulation of reactive oxygen species, up-regulated LC3 protein expression and increased mRNA levels of ATG3 , ATG5 and ATG7 . Annexin V-FITC staining assay revealed that AFB1 treatment (50 μM) resulted in oocyte early apoptosis, which was confirmed by increased Bak , Bax , Bcl-xl mRNA levels. Collectively, our results suggest that AFB1 disrupts porcine oocyte maturation through changing epigenetic modifications as well as inducing oxidative stress, excessive autophagy and apoptosis.

Description: Epidemiological studies have demonstrated a close association between infection with Helicobacter pylori ( H.pylori ) and the development of gastric carcinoma. Chronic H.pylori infection increases the frequency of mutation in gastric epithelial cells. However, the mechanism by which infection of H.pylori leads to mutation in gastric epithelial cells is unclear. We suspected that components in H.pylori may be related to the mutagenic response associated with DNA alkylation, and could be detected with the Ames test using a more sensitive strain for alkylating agents. Our investigation revealed that an extract of H.pylori was mutagenic in the Ames test with Salmonella typhimurium YG7108, which is deficient in the DNA repair of O 6 -methylguanine. The extract of H.pylori may contain methylating or alkylating agents, which might induce O 6 -alkylguanine in DNA. Mutagenicity of the alkylating agents N -methyl- N -nitrosourea (MNU) and N -methyl- N '-nitro- N -nitrosoguanidine in the Ames test with S.typhimurium TA1535 was enhanced significantly in the presence of the extract of H.pylori. The tested extracts of H.pylori resulted in a significant induction of micronuclei in human-derived lymphoblastoid cells. Heat instability and dialysis resistance of the extracts of H.pylori suggest that the mutagenic component in the extracts of H.pylori is a heat-unstable large molecule or a heat-labile small molecule strongly attached or adsorbed to a large molecule. Proteins in the extracts of H.pylori were subsequently fractionated using ammonium sulphate precipitation. However, all fractions expressed enhancing effects toward MNU mutagenicity. These results suggest the mutagenic component is a small molecule that is absorbed into proteins in the extract of H.pylori , which resist dialysis. Continuous and chronic exposure of gastric epithelial cells to the alkylative mutagenic component from H.pylori chronically infected in the stomach might be a causal factor in the gastric carcinogenesis associated with H.pylori .

Description: The buccal micronucleus cytome (BMCyt) assay is a minimally invasive approach for measuring DNA damage, cell proliferation, cell differentiation and cell death in exfoliated buccal cells. The main limitation for its use is the lack of knowledge about inter- and intra-laboratory variability in scoring micronuclei and other end points included in the cytome approach. In order to identify the main sources of variability across the BMCyt biomarkers, a scoring exercise was carried out between three experienced laboratories using the same set of slides and an identical set of detailed scoring criteria and associated images for the different end points. Single batches of slides were prepared from pooled samples of four groups of subjects characterised by different frequencies of cell types and micronuclei, namely Down syndrome patients, head and neck cancer patients undergoing radiotherapy and two age- and gender-matched control groups. A good agreement among the laboratories in the identification of normal differentiated cells and of micronuclei was obtained. A 3-fold and 20-fold increase in the frequency of micronucleated cells and micronuclei in differentiated cells of Down syndrome patients and in cancer patients, respectively, compared to matched controls, was a consistent result in the three laboratories. The scores of other cell types and nuclear anomalies, such as basal, binucleated, condensed chromatin and karyorrhectic cells showed significant disagreement between and within laboratories indicating that their evaluation using the current visual scoring protocol does not yield robust results for these parameters. The guidelines for BMCyt assay application could be improved by combining the anomalies associated with cell death (condensed chromatin and karyorrhectic cells) in a single category and by defining more stringent criteria in classifying basal cell, binucleated cells and buds.

Description: Deregulation of Wnt/β-catenin signalling plays an important role in the pathogenesis of colorectal cancer. Interestingly, this pathway has been recently implicated in transcriptional control of cytochrome P450 (CYP) family 1 enzymes, which are responsible for bioactivation of a number of dietary carcinogens. In the present study, we investigated the impact of inhibition of Wnt/β-catenin pathway on metabolism and genotoxicity of benzo[a]pyrene (BaP), a highly mutagenic polycyclic aromatic hydrocarbon and an efficient ligand of the aryl hydrocarbon receptor, which is known as a primary regulator of CYP1 expression, in cellular models derived from colorectal tumours. We observed that a synthetic inhibitor of β-catenin, JW74, significantly increased formation of BaP-induced DNA adducts in both colorectal adenoma and carcinoma-derived cell lines. Using the short interfering RNA (siRNA) targeting β-catenin, we then found that β-catenin knockdown in HCT116 colon carcinoma cells significantly enhanced formation of covalent DNA adducts by BaP and histone H2AX phosphorylation, as detected by 32 P-postlabelling technique and immunocytochemistry, respectively, and it also induced expression of DNA damage response genes, such as CDKN1A or DDB2 . The increased formation of DNA adducts formed by BaP upon β-catenin knockdown corresponded with enhanced production of major BaP metabolites, as well as with an increased expression/activity of CYP1 enzymes. Finally, using siRNA-mediated knockdown of CYP1A1, we confirmed that this enzyme plays a major role in formation of BaP-induced DNA adducts in HCT116 cells. Taken together, the present results indicated that the siRNA-mediated inhibition of β-catenin signalling, which is aberrantly activated in a majority of colorectal cancers, modulated genotoxicity of dietary carcinogen BaP in colon cell model in vitro , via a mechanism involving up-regulation of CYP1 expression and activity.

Description: We undertake a systematic study of asymptotically hereditarily aspherical (AHA) groups, the class of groups introduced by Tadeusz Januszkiewicz and the second author as a tool for exhibiting exotic properties of systolic groups. We provide many new examples of AHA groups, also in high dimensions. We relate the AHA property with the topology at infinity of a group, and deduce in this way some new properties of (weakly) systolic groups. We also exhibit an interesting property of boundaries at infinity for a few classes of AHA groups.

Description: We show that certain geometrically defined higher codimension cycles are extremal in the effective cone of the moduli space ${\overline {\mathcal M}}_{g,n}$ of stable genus $g$ curves with $n$ ordered marked points. In particular, we prove that codimension 2 boundary strata are extremal and exhibit extremal boundary strata of higher codimension. We also show that the locus of hyperelliptic curves with a marked Weierstrass point in ${\overline {\mathcal M}}_{3,1}$ and the locus of hyperelliptic curves in ${\overline {\mathcal M}}_4$ are extremal cycles. In addition, we exhibit infinitely many extremal codimension 2 cycles in ${\overline {\mathcal M}}_{1,n}$ for $n\geq 5$ and in ${\overline {\mathcal M}}_{2,n}$ for $n\geq 2$ .