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Simple Elastic Network Models for Exhaustive Analysis of Long Double-Stranded DNA Dynamics with Sequence Geometry Dependence
Simple elastic network models of DNA were developed to reveal the structure-dynamics relationships for several nucleotide sequences. First, we propose a simple all-atom elastic network model of DNA that can explain the profiles of temperature factors for several crystal structures of DNA. Second, we propose a coarse-grained elastic network model of DNA, where each nucleotide is described only by one node. This model could effectively reproduce the detailed dynamics obtained with the all-atom elastic network model according to the sequence-dependent geometry. Through normal-mode analysis for the coarse-grained elastic network model, we exhaustively analyzed the dynamic features of a large number of long DNA sequences, approximately bp in length. These analyses revealed positive correlations between the nucleosome-forming abilities and the inter-strand fluctuation strength of double-stranded DNA for several DNA sequences.
I Introduction
Elastic network models of proteins, including all-atom models(1); (2) and coarse-grained models(3); (4); (5); (6); (7); (8); (9) represent some of the simplest and most powerful types of theoretical models that can accurately reveal structure-dynamics relationships and the mechanisms underlying a protein's functional activities (10); (11); (12); (13); (14). Such models have also been widely employed to accurately reproduce the temperature factors on the crystal structure of a protein via normal-mode analysis. These models can thus demonstrate the large and slow deformations of proteins that are essential for protein functions, but remain difficult to demonstrate via all-atom molecular dynamics simulations.
Along with proteins, DNA is the most important biomolecule for the activities of all living organisms. Recent developments in molecular biology have revealed that DNA contains several functional regions. Therefore DNA is no longer considered to have the sole function in the storage of genetic information, but is now known to be actively involved in gene regulation(15); (16); (17); (18), insulator activity(19); (20); (21); (22); (23); (24), and in the construction of chromosomal architectures(26); (27); (25); (28) such as heterochromatins and topologically associated domains through nucleosome formation and protein bindings(29); (30); (31); (32); (33). The functional behavior of each strand of DNA is determined not only by chemical aspects of the nucleotides and base pairs but also by its physical characteristics such as the structure and dynamics of the nucleotide sequences in each functional region. However, comprehensive understanding of the physical properties of nucleotide sequences lags far behind the knowledge accumulated of their biochemical aspects (15); (16); (26).
Since the last century, much progress has been made in revealing the physical aspects of DNA using all-atom normal-mode analysis(34); (35) and molecular dynamics simulations(36); (37); (38); (39); (40); (41); (42). Several coarse-grained models of DNA (and RNA) have also been proposed. Some of these describe the detailed shape of each nucleotide using three or more particles(43); (44); (45); (46); (47); (48); (49), whereas others describe each base pair by simply one or two particles(50); (52); (51); (53); (54). Molecular dynamics simulations and normal-mode analysis of these models have identified the flexibilities, nucleosome-forming abilities, and zip-unzip transitions of the double helices of some specific DNA sequences from tens to a few hundred base pairs in length. Although these methods have proven to be very powerful for the analysis of the physical aspects of DNA, molecular dynamics simulations are not suitable for conducting exhaustive analyses of several sequences simultaneously owing to the high computational costs of such extensive simulations; for example, analyses of whole genomic and whole possible sequences. Moreover, the normal-mode analysis of all-atomic models of long DNA sequences is also computationally heavy.
Alternatively, data-driven methods have been proposed for determining the mechanical properties of DNA with respect to the helical parameters and local flexibilities of base pairs from X-ray crystal structure analysis and all-atom molecular dynamics (55); (56); (57); (58); (59); (60). These methods also seem to be powerful and can be applied to the analysis of the mechanical properties of several DNA sequences simultaneously. However, the methods thus far proposed have focused on static and local mechanical properties. Therefore, they are not particularly useful for the study of the functional contribution of the dynamic and correlated motions of DNA.
The objective of this study was to construct a simple coarse-grained elastic network model of DNA to allow for exhaustive analysis of the dynamic correlated motions of several long DNA sequences. For this purpose, we first constructed a simple all-atom elastic network model of short DNA sequences based on the method introduced by Tirion(1) for the modeling of protein dynamics. We confirmed that this model could accurately reproduce the temperature factors of some DNA fragments from data obtained with X-ray crystal structure analysis.
Second, we developed a simple coarse-grained elastic network model of long DNA sequences, where each nucleotide is described by only one node. We confirmed that this simplified model has lower computational costs but can nonetheless reproduce the nucleotide sequence-dependent dynamics revealed by the all-atom elastic network model.
Finally, through the normal-mode analysis of this coarse-grained model, we conducted an exhaustive analysis of the general features of the sequence-dependent structure-dynamics relationships among several DNA sequences. We specifically focused on the dynamic properties of a large number of long DNA sequences, approximately bp in length (with respect to the length of the nucleosome-forming regions), for the genomes of some model organisms as well as for random sequences with several A, T, C, G ratios. Through these analyses, we found that the dynamic aspects of DNA are highly influenced by their sequences, and found positive correlations between the nucleosome-forming abilities and inter-strand fluctuations of double-stranded DNA.
In particular, we focus on the geometry dependencies of the dynamics of several DNA sequences, since a recent study demonstrated that the geometry of DNA sequences has a dominant contribution to their mechanical features(47). In the following arguments, for simplicity, we use "sequence-dependent" to mean "dynamics that depend on sequence geometry".
Ii Models and Methods
ii.1 Basic structures of Elastic Network Models of DNA
In order to construct elastic network models of DNA, the basic DNA structure first needs to be determined. In the following arguments, we construct two types of models: i) a simple all-atom elastic network model (AAENM) to reproduce the characteristics of the crystal structures of DNA, and ii) a simple coarse-grain elastic network model (CGENM) to reproduce the characteristics of the AAENM. We obtained the basic DNA structures in the following two ways for the respective purposes of constructing each model.
For construction of the AAENM, we employed the atom coordinate sets of several naked DNA crystal structures included in the Protein Data Bank (PDB) as the given basic structures of the model. We used DNA crystal structures containing only DNA and water molecules under different conditions of crystallization, where all temperature factors are given as positive values (Table 1). The suitability of the model was evaluated by comparisons of the temperature factors obtained between the model and those obtained from X-ray crystal structure analysis.
The objective of our constructed arguments was to unveil the sequence-dependent dynamic features of several long DNA sequences simultaneously. In recent crystal structure analysis, only shorter DNA sequences (i.e., less than bp in length) were studied. On the other hand, several types of helical parameter sets, base pair parameters, and base step parameter sets have been proposed through experiments or molecular dynamics simulations(36); (61); (62); (63); (64); (65); (66)(Table 2, S1 Table). Here, we used the application X3DNA(67) to obtain the coordinates of each atom for any sequence from these helical parameters, and used these parameters to construct the AAENMs and CGENMs of longer DNA sequences (for detailed methods of the generation of the atom coordinates, see Lu and Olson (67)). In the following arguments, we compare the characteristics of the AAENM and CGENM constructed by X3DNA from three different helical parameter sets: i) a parameter set obtained by an in vitro experiment and X-ray crystal structure analysis(48); (61); (62); (63) (Table 2), ii) a parameter set obtained only by the X-ray crystal structure analysis, and iii) a parameter set obtained by all-atom molecular dynamics simulations(58); (65); (66) (S1 Table).
ii.2 All-atom Elastic Network Model of DNA
A simple all-atom elastic network model of double-stranded DNA was constructed based on the model proposed by Tirion(1). In this model, we regard all of the atoms of given DNA sequences as the nodes comprising the elastic network. For the analysis of the crystal structures of DNA involving water molecules, we also regard the oxygen atoms of water around DNA as the nodes of the elastic network. We define the mass and position of atom as and (), respectively. The potential of all atoms is given as
Here, is the position of atom of the basic DNA structure, as described above.
The first term indicates the interaction potential among atoms that are spatially closed in the basic DNA structure (Fig. 1(a)). Here, refers to the Van der Waals radius of atom , is an arbitrary cut-off parameter that models the decay of interactions with distance, and indicates the Heaviside function, where () for (). We assume , which is empirically considered to be an appropriate range for biomolecules, at least for proteins(1). The results of the arguments presented in this paper were qualitatively unchanged for the appropriate range of . In the crystal structures of DNA, the motions of water molecules are also restricted by the crystal packing. Thus, for all atoms, we assume that spatially closed pairs of atoms are connected by linear springs with their respective natural lengths. The elastic coefficient () is a phenomenological constant, which is assumed to be the same for all interacting pairs.
The second term indicates the boundary effects of each DNA and water molecule in each DNA crystal structure. This term plays a crucial role for the analysis of the fluctuations of the crystal structure of DNA, such as temperature factors, since the fluctuations of the nucleotides at the edges of DNA and water molecules are restricted due to the following facts.
In the crystal of DNA, the motion of edges at upper and lower streams of each DNA segment (left and right edges in Fig. 1(a)) is influenced by atoms of other adjacent segments in the long-axis direction of DNA segments. Moreover, the motions of water molecules around each DNA segment are influenced by atoms of other adjacent water molecules or DNA segments in the direction perpendicular to the longitudinal axis of the DNA segment. Thus, we need to consider the second term of Eq. (1) to implement such effects, where indicates the strength of such effects for atoms in the edge nucleotides of each DNA and water molecule, respectively.
Remarkably, as shown in the Results section, the distributions of the temperature factor of atoms exhibited various patterns from the same sequence of crystallized DNA (S1 Fig and S2 Fig) owing to the dependency on the conditions of crystallization. Therefore, in order to compare the characteristics of the present DNA model to those of the crystal structure of DNA, appropriate values of need to be assigned to the atom that belongs to the edge base pairs and water molecules. For simplicity, we assign and to the atoms at the edge base pairs and the water molecules, respectively, and otherwise.
It is noted that the internal structure of the crystal of DNA is spatially anisotropic. Thus, it is reasonable to assume that the interactions among local parts of the crystal of DNA exhibit different strengths in different directions. Accordingly, in general, the strengths of the restrictions of atoms belonging to the edge of DNA differ from those of water molecules. Thus, we assume that and are different values (Table 1).
For simplicity, the mass of each atom is assumed as a constant value (, is Avogadro's number). However, we confirmed that the results were almost identical when using the precise masses of the atoms.
ii.3 Coarse-grained Elastic Network Model of DNA
A simple coarse-grained elastic network model of double-stranded DNA, where each nucleotide is described as one node, was constructed as follows (Fig. 1(a)). We define the coordinate of the C1' carbon of nucleotide , (), as the position of nucleotide , and regard the motion of the C1' carbon as that of the nucleotide. Here, we assume that the mass of the C1' carbon obeys . The potential of all nucleotides is given as
Here, is the position of nucleotide of the basic structure of DNA, as defined above. We assume that nucleotides and belong to the same base pair. For nucleotide , the sum is restricted to the pair of nucleotides in the same base pair (), in the neighboring base pairs (), in the next neighboring bases pairs (), and in the next-next neighboring bases in another strand () (Fig. 1(b)). The elastic coefficient () is a phenomenological constant that is assumed to be the same for all interacting pairs. This model is considered as a simplified version of previously proposed one-site-per-nucleotide models(50); (53).
ii.4 Normal-mode Analysis
An overview of the theory of normal-mode analysis is provided in several recent studies(1); (2); (3); (4); (10); (11); (12); (13); (5); (6); (7). Thus, we here briefly show the results of this analysis. For this analysis, we define (, ) as a -dimensional position vector, and as the position vector of the basic structure. Here, for the AAENM and for the CGENM. The motions of small deviations of from , obey
where and () are the -th largest eigenvalue and its eigenvector of the Hessian matrix as
We assume that the system is in thermodynamic equilibrium with temperature . Thus, the amplitude is given as
with Boltzmann constant . Using this solution, the mean square fluctuation of the -th atom in the AAENM () is obtained as
with Boltzmann constant, and the temperature factor is displayed as . Here, represents the temporal average.
For the CGENM, we define the mean square fluctuation of the -th nucleotide () as . To consider the motion of the -th nucleotide in the AAENM, we define the average nucleotide motions as . Using this vector, we define the mean square fluctuation of the -th nucleotide as . For the motif of the -th nucleotide ( sugar, base, and phosphoric acid), the average temperature factor of the motif () is defined as the average of belonging to each motif of each nucleotide, .
ii.5 Treatment of the Temperature Factor in X-ray crystal Structure Analysis
In order to evaluate the validity of the AAENM, we measured the correlation coefficient between the profile of the temperature factor obtained from PDB data (via X-ray crystal structure analysis) and that obtained from the AAENM based on this crystal structure. It is noted that the temperature factor profiles for some of the PDB data often include unnaturally large or small values. Thus, the correlation coefficients were estimated using data excluding such outliers. In the present evaluations, the value was considered as an outlier if , where and are the average and standard deviation of , respectively, and is used based on the standard arguments of statistics.
ii.6 Evaluations of Anisotropic Fluctuations of DNA
We also focused on the relationships between the fluctuations of each nucleotide in the AAENMs and CGENMs in the directions parallel to the base pair axis, parallel to the helix axis, and vertical to both the base pair and helix axes.
Here, we name the nucleotides in one and the other strand constructing the -th base pair as the -th and -th nucleotide. We define the position vectors of the C1' atoms belonging to the -th and -th nucleotides as and , and consider
(Fig. 1(c)). It is noted that and are not orthogonal in general; however, we confirmed that the angles of these vectors were always sufficiently close to rad for each .
Using these vectors, the mean square fluctuations of the -th and -th nucleotides of the AAENM in the directions parallel to the base pair axis are defined as and , those parallel to the helix axis are defined as and , and those in the torsional direction are defined as and , respectively. The mean square fluctuations of the -th and -th nucleotides of the CGENM in these same directions are obtained by and , and , and and , respectively. Moreover, we consider the mean square fluctuation of the relative base position of each base pair of the CGENM in the three directions listed above given by , , and , respectively.
ii.7 Evaluations of the Overall Geometry of DNA
The overall geometry of each modeled DNA molecule is characterized by the ratios among the square root of the three principal components of the populations of atoms, , , and (). Here, , , and are obtained as eigenvalues of the covariant matrix
where , is the position of -th atom, is the position of the center of mass of a given DNA molecule, and indicates the average for all s. We evaluate the overall geometry of a given DNA molecule using the linearity and the line symmetry with respect to the -axis . Here, is large when the DNA is straightened, and is large (small) when the DNA forms a three (two)-dimensional curve with wide (flat) envelope.
ii.8 Evaluations of Correlations Among the Results of AAENM, CGENM, and Experiments
We employ Pearson's correlation coefficients, , to evaluate the correlations among the profiles of temperature factors and several anisotropic fluctuations of atoms obtained by AAENM, CGENM, and experiments.
Iii Results and Discussion
iii.1 Comparisons of Fluctuations between the AAENM and the Crystal Structure of DNA
The fluctuations of atoms of the AAENMs of several short DNA sequences were measured with normal-mode analysis. Here, the basic structures of the DNAs are given by the crystal structures of the naked DNAs with the following PDB IDs: 1BNA, 7BNA, 9BNA, 1D91, 1DC0, 122D, 123D, 181D, and 330D (Table 1(68); (69); (70); (71); (72); (73); (74); (75)). To confirm the validities of the AAENMs, the correlations between the distribution profiles of the temperature factor of atoms () and the average temperature factor of the motifs () in the crystal structures and those in the corresponding AAENMs were measured. In the following arguments, we employ the optimal values of , , and for each crystal structure (Table 1), which were manually found to maximize of between the results of the crystal structure analysis and those of the AAENM.
By choosing the appropriate conditions for the atoms of the two edge base pairs and water molecules ( and ) for each PDB ID, of each AAENM exhibited a similar profile to that of the crystal structure with a significant correlation coefficient (Fig. 2(a), Table 1, and S1 Fig). Therefore, the AAENM seemed to reproduce the overall structure of the temperature factor profile of each crystal structure well, although the detailed profiles among atoms showed some deviations.
Furthermore, we focused on the average temperature factor for the motifs, , as recently discussed (35). We found that the obtained with the AAENMs could accurately reproduce those of the corresponding crystal structures with high correlation coefficients , where was obtained for most cases (Fig. 2(b), Table 1, and S2 Fig). These results demonstrate that the AAENM is a suitable model for describing the sequence-dependent fluctuations of the nucleotide motifs of several double-stranded DNA sequences, despite the simplicity of model construction and its easy implementation. Moreover, these results show that the sequence-dependent forms of DNA have a dominant contribution to their overall flexibilities and fluctuations.
Nevertheless, the present AAENM could not accurately reproduce well for some crystal structures. These deviations are considered to have arisen from the following primary assumption: we only considered the effects of the restrictions by the packing of DNAs in crystal form for two edge base pairs and water, whereas such effects have an influence on all atoms. Therefore, obtaining and incorporating more detailed knowledge of the restrictions of bulk sequences caused by crystal packing should help to achieve a more accurate reproduction of the molecular fluctuations for all cases.
iii.2 Comparison between the AAENMs and CGENMs of DNA
The main objective of this study was to unveil the sequence-dependent dynamic correlated motions of several long DNA sequences. We next describe these dynamics of DNA sequences with longer lengths than considered in the previous subsection. For this purpose, we also constructed a coarse-grained model, which is often useful for focusing on the slow and large-scale dynamics of molecules that essentially influence their function. Thus, to propose a coarse-grained model of long double-stranded DNA, we evaluated whether the CGENM proposed provides an appropriate coarse-grained model of the present AAENM.
We performed normal-mode analysis of the AAENM and corresponding CGENM for randomly chosen -bp sequences, and compared the mean square fluctuations of the -th nucleotide ( and ) in the directions parallel to the base pair axis ( and ), parallel to the helix axis ( and ), and in the torsional direction ( and ). Here, is employed, as in the previous study(1), and is assumed, which was manually found to provide the best fit of fluctuation profiles between AAENM and CGENM. Here, the overall fluctuation profiles of CGENM are independent of the value of since influences only on the absolute values of fluctuations. Independent of the sequences and employed helical parameters, we found that the fluctuations of each nucleotide in several directions were very similar between the AAENMs and CGENMs when these models are constructed with the same helical parameters, with average correlation coefficients (Fig. 3, Table 3, S2 Table, S3 Fig, and S4 Fig).
It is noted that the present CGENM contains only one node per nucleotide, whereas the AAENM contains nodes (atoms) per nucleotide. This fact demonstrates that the computational costs of the CGENMs are much lower than those of the AAENMs, although the accuracies of the obtained statistical aspects are almost identical between these two models. Thus, this CGENM could be used for exhaustive analysis and comparisons of the dynamic features of several sequence-dependent DNAs related to protein binding affinities, functions of transcription regulation sequences, and nucleosome positioning(16); (26); (27); (17); (18); (19); (20); (21); (22); (23); (24); (46); (55); (56); (57); (58); (59); (60). In the next subsection, we provide an example of such an analysis to determine the relationships between the nucleosome-forming abilities of several double-stranded DNA sequences and their inter-strand dynamic features.
iii.3 Exhaustive Analysis of the Sequence-dependent Behaviors of -bp DNAs with the CGENM
Nucleosome positioning is important not only for compacting DNA but also for appropriate gene regulation. Several recent studies have been performed for genome-wide nucleosome mapping and the identifications and predictions of nucleosome-forming and -inhibiting sequences for some model organisms(57); (59); (76); (77); (78); (79); (80).
As an example of the applications of the CGENM to an exhaustive analysis of the sequence-dependent behavior of DNA, we compared the dynamic features of DNA sequences of bp that were predicted as nucleosome-forming or nucleosome-inhibiting sequences in the genome of budding yeast Saccharomyces cerevisiae (5000 forming sequences and 5000 inhibiting sequences of bp)(57), nematode Caenorhabditis elegans (2567 forming sequences and 2608 inhibiting sequences of bp), Drosophila melanogaster (2900 forming sequences and 2850 inhibiting sequences of bp), and Homo sapiens (2273 forming sequences and 2300 inhibiting sequences of bp)(59). The histograms of the average relative fluctuations of DNAs for the three directions , , , , , , , and ( indicates the average for all s.) for the nucleosome-forming sequences and the nucleosome-inhibiting sequences are shown in Fig. 4 and S5 Fig S7 Fig. Here, we employed the helical parameter set (i) (Table 2) that was used in the coarse-grained molecular dynamics simulations by Freeman et al., which exhibited consistent results to some experiments(48); (61); (62); (63).
The histograms for budding yeast showed that that nucleosome-forming sequences tend to exhibit larger fluctuations in several directions compared to the inhibiting sequences. In particular, the histogram of for the nucleosome-forming sequences showed a clear shift in the direction toward larger values compared to that for the nucleosome-inhibiting sequences (Fig. 4). For the other organisms, most of the histograms showed few differences between the nucleosome- forming and -inhibiting sequences. However, similar to the case of yeast, the distribution of for the nucleosome-forming sequences shifted largely in the direction of larger values compared to that for the nucleosome-inhibiting sequences in these organisms (S5 Fig S7 Fig). These results indicate that the nucleosome-forming ability is highly correlated to the fluctuations of the inter-strand distances of DNAs, in which sequences with larger fluctuations tend to form the nucleosome.
Similar to the previous arguments, we compared the dynamical features among the random -bp DNA sequences varying in average GC-contents; the histograms, averages, and standard deviations of , , , , and were measured from 10,000 random sequences for each GC content (Fig. 5 and S8 Fig). In this case, exhibited a minimum at a GC content of , which indicates that the AT-rich sequences tend to be more rigid than the GC-rich sequences. However, the fluctuations of sequences consisting of only A or T were as large as those of the GC-rich sequences. Moreover, the GC content dependencies of , , , and showed different characteristics for GC contents larger or smaller than . In particular, the results for were similar for cases with a large GC content () but monotonically decreased with a GC content with little variance. A recent experimental study showed that the probability of nucleosome formation tends to increase with increases in the GC content ratio (81). Thus, the present results indicate that sequences with larger fluctuations of inter-strand distances tend to form the nucleosome, which is consistent with the results described above from the analysis of the genomes of the four model organisms.
Finally, we focus on the relationships between the overall geometries and fluctuations of the considered DNA sequences. The overall geometries of several DNA sequences, the nucleosome-forming and -inhibiting DNA sequences for four model organisms and random DNA sequences with different GC contents, were evaluated using scatter plots of (linearity) and (line symmetry) (Fig. 6 and S9 Fig). It is noted that and are highly correlated. The average, standard deviation, and distributions of and exhibited slight but not significant deviations between the nucleosome-forming and -inhibiting sequences.
For the random sequences, the average value of exhibited similar variations to with an increase in GC content. In particular, both values decreased with increases in GC content for GC contents , whereas they increased with increases in GC content for GC contents (Fig. 5(c) and Fig. 6(c)). In fact, and were highly correlated for random DNA sequences, regardless of their CG content. The Pearson correlation coefficient for the 110,000 sequences analyzed above with GC contents was . It is noted that showed large dispersion for each GC content, and there were significant overlaps among distributions with different GC contents (Fig 6(b) and 6(c)). This fact indicates that different DNA sequences can often show similar geometries, and such sequences also tend to show similar overall fluctuations. On the other hand, the fluctuations of inter-strand distances that may correlate to the nucleosome-forming ability did not correlate significantly to either or , with Pearson correlation coefficients of and . This fact indicates that the nucleosome-forming ability of DNA sequences are not only determined by the overall DNA geometries but also by their dynamic properties.
Iv Summary and Conclusion
In this study, simple elastic network models of double-stranded DNAs were developed in order to perform an exhaustive analysis of several sequence-dependent dynamical features. First, we constructed a simple all-atom elastic network model that could reproduce the fluctuations of the motifs of each nucleotide (sugar, phosphoric acid, and bases) of several crystal structures of short DNA sequences. Second, we proposed a simple course-grained elastic network model that could reproduce the dynamic features of the long DNA sequences obtained by the all-atom elastic network model. Through exhaustive analysis of the dynamic features of several DNA sequences with normal-mode analysis of the presented coarse-grained elastic network model, we found that the dynamic aspects of DNA are highly influenced by the properties of nucleotide sequence such as GC content. We also found that the nucleosome-forming abilities of double-stranded DNA exhibited positive correlations with their sequence-dependent inter-strand fluctuations.
In the present study, we demonstrated the sequence-dependent dynamic features for several -bp DNA sequences to evaluate the relationships between the nucleosome-forming abilities and DNA dynamics. Of course, DNA sequences longer than bp can also be analyzed using the presented course-grained model. Moreover, course-grained molecular dynamics simulations can be performed to consider the large deformations of DNA, such as formation of a super helix and nucleosome that are the basic structures of higher-order chromosome architectures, using the presented elastic network models with the excluded effect of the volume of each atom or each nucleotide. We are currently conducting these molecular dynamics simulations, and the results will be reported in the future. We did not consider the effects of solvents such as temperature and salt concentrations in the present elastic network models. Therefore, we are also planning to attempt modifications of the models so that several solvent conditions can be incorporated in future work.
The author is grateful to S. Tate and Y. Murayama for fruitful discussions.
Author contributions
Conceived the study: AA, NS, HN. Designed the models: NS, AA, SI. Performed the analysis: SI, AA. Wrote the paper: AA, NS, HN.
Supporting Information
S1 Table
S2 Table
Comparisons between CGENM and AAENM. Average and standard deviation of the correlation coefficients of 500 samples of random -bp sequences between and , and , , and , and and . Helical parameter sets (ii) and (iii) (S1 Table) were used.
S1 Fig
Temperature factor of each atom. The temperature factor of each atom obtained by the AAENMs (black curve) and X-ray crystal structure analysis (gray (red) curve) of typical double-stranded DNAs obtained from PDB ID (a) 7BNA, (b) 9BNA, (c) 1D91, (d) 1DC0, (e) 122D, (f) 123D, (g) 181D, and (h) 330D. Parameters , , and are given in S1 Table.
S2 Fig
Average temperature factor of motifs. Average temperature factor of motifs obtained by the AAENMs (black curve) and X-ray crystal structure analysis (gray (red) curve) of typical double-stranded DNAs obtained from PDB ID (a) 7BNA, (b) 9BNA, (c) 1D91, (d) 1DC0, (e) 122D, (f) 123D, (g) 181D, and (h) 330D. Parameters , , and are given in S1 Table.
S3 Fig
Comparisons between AAENM and CGENM. Comparisons of the fluctuations between each nucleotide in the AAENM (black curves) and CGENM (gray (red) curves) for a typical random -bp DNA sequence (5' - AGTGGTAAGGCATGGTTCTCGAATCTCGGTTTATTTACACTGCTGCTCCA - 3'). (a) and , (b) and , (c) and , and (d) and using helical parameter set (ii) S2 Table.
S4 Fig
Comparisons between AAENM and CGENM. Comparisons of the fluctuations between each nucleotide in the AAENM (black curves) and CGENM (gray (red) curves) for a typical random -bp DNA sequence (5' - ATATGCTGTAGAGCGTCCCGTCCGCGCGTTGTGGTTTTTTCGGTGCTCTA - 3'). (a) and , (b) and , (c) and , and (d) and using helical parameter set (iii) S1 Table.
S5 Fig
Histograms of the average fluctuations inCaenorhabditis elegans. Histograms of the average fluctuations of (a) and , (b) and , (c) and , and (d) and for nucleosome-forming and nucleosome-inhibiting sequences of the nematode Caenorhabditis elegans ( bp). Helical parameter set (i) (Table 2) was used.
S6 Fig
Histograms of the average fluctuations inDrosophila melanogaster. Histograms of the average fluctuations of (a) and , (b) and , (c) and , and (d) and for nucleosome-forming and nucleosome-inhibiting sequences of Drosophila melanogaster ( bp). Helical parameter set (i) (Table 2) was used.
S7 Fig
Histograms of the average fluctuations inHomo sapiens. Histograms of the average fluctuations of (a) and , (b) and , (c) and , and (d) and for nucleosome-forming and nucleosome-inhibiting sequences of Homo sapiens ( bp). Helical parameter set (i) (Table 2) was used.
S8 Fig
Histograms of the average fluctuations with different GC contents. Histograms of (a) , (b) , and (c) for 10,000 samples of random -bp sequences with different average GC contents. Helical parameter set (i) (Table 2) was used.
S9 Fig
Overall geometries of long DNA sequences in model organisms. Scatter plots of and for nucleosome-forming and -inhibiting sequences of (a) Caenorhabditis elegans ( bp), (b) Drosophila melanogaster ( bp), and (c) Homo sapiens ( bp). Helical parameter set (i) (Table 2) was used.
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Issues in global news translation (case of Latvia in Polish media)The purpose of this paper is to discuss the process of translation of the news made in specific language-cultural context. Theoretical basis of the analysis are critical discourse analysis and postcolonial studies as well as translation and media studies. Empirical part of the paper aims at reconstruction and qualitative analysis of a complex media fact (as opposite to actual fact) of "commemoration of Nazi murderers within former Nazi concentration camp." Possible process of creation and circulation of the news is reconstructed and two companion phenomena are analysed. Those are: hidden ideologization of the news in Russian media as well as recontextualisation of the same news in Polish media.
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Petition update
BREAKING: COK Video of Cruelty to Chickens Prompts Groundbreaking Trials & Convictions!
Aug 30, 2017 — In a groundbreaking series of trials and plea agreements prompted by Compassion Over Killing's footage filmed inside several Tyson Foods broiler breeder facilities in Virginia, nine former Tyson employees have been convicted on 22 counts of animal cruelty. These convictions--the first-ever for broiler breeders--are a monumental victory in achieving justice for farmed animals! But we can't stop now: Share our petition TODAY to urge Tyson to address the suffering of the nearly 2 billion birds it raises and kills each year!
COK's investigation documented Tyson workers punching and kicking live birds, and birds being crushed to death by transport crates and run over by forklifts. The video, which offered the first investigative look inside broiler breeder factory farms, also revealed for the first time on hidden camera, the barbaric use of "nose bones"--in which a dull plastic rod is stabbed through male breeder birds' nostrils to restrict food intake. Now, COK's unprecedented footage has led to the conviction of a former Tyson worker for engaging in this cruel practice, among multiple other charges.
"Nose bones" are flying the coop, as Tyson and several other major chicken producers have stopped using them since the release of COK's video, but Tyson has yet to address the root problem: This poultry giant still breeds birds to grow so unnaturally large, so fast that they are starved so they'll live long enough to breed.
In a landmark moment for broiler breeders, these convictions are sending a clear message that "those who commit cruelty to animals will be held accountable for their actions," as Virginia Attorney General Mark Herring said. Yet millions of birds are still suffering in darkness at Tyson. Help us end this suffering: Share our petition (http://bit.ly/EndTysonAbuse) and tag @TysonFoods on social media today!
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Double Helix Bead Crochet Necklace & Bracelet with Darla J. Fanton
Read Darla J. Fanton's bio here!
Bead loops spiral around a central core of crochet thread in this lovely 21-inch long necklace and matching bracelet. It is hard to believe that the very simplest of crochet stitches are used to create these projects, but it's true! Those students already familiar with crochet should be able to leave class wearing their necklace. Those new to crochet may wish to start with the bracelet.
Kits will be available in assorted colors and will make one necklace and one bracelet with sufficient thread for additional projects.
Materials Provided:
Kit includes:
crochet thread pre-strung with glass beads
toggle clasps
crochet hook
Materials to bring:
Tape measure
Portable Task Light / Magnifier (Optional)
Skill level: Novice - No experience necessary.
Material Fee: $25
Class Length: 3 hours
Class size: 15 students
Spaces Available: 10
This class will be held: Saturday, April 20th, 2013 Afternoon - 1pm - 4pm
Please read instructions carefully on the Workshop Page before signing up!
©2012 - 2013 LavenderSheep - Yvonne Ellsworth


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Game 1 of the NBA finals is behind us. The memes are out, mostly making fun of the Golden State Warriors while praising the Toronto Raptors, Pascal Siakam (aka Spicy P) and even Drake. More
Numbers, titles, awards – they're all measurable, tangible, objective. That big word, legacy? A bit more difficult to grasp or agree on. By enhancing the myth of himself and the Golden State Warriors, Stephen Curry is puncturing holes all through the legacy of Kevin Durant. IRobot Discovery Roomba Robotic Vacuum Cleaner Model 4210 Original Box Extras
Some statistics mean something, others less. But sometimes the mundane and borderline useless becomes handy, so perhaps the Toronto Raptors can draw some confidence from the title of this article. More
While there was more to the 2018-2019 Premier League season than just the historically bountiful title race between Manchester City and Liverpool, in the years that will come it will be what we remember from a season filled with big storylines and lessons that deserve to be learned. More
It took a wonderful goal by Vincent Kompany for Manchester City, with all its money and talent, to overcome a stubborn and dangerous Leicester side. The defending champions are now one win away against Brighton from claiming the 2019 Premier League title. LUNA Weighted Blanket (15 lbs - 60x80) 100% Organic Cooling Cotton &...
It's the third year for Russell Westbrook without Kevin Durant, trying to lead the Oklahoma City Thunder past the first round in the NBA playoffs. As the memes spreading through the Internet suggest, it's not going too well. More
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The race continues. A day after Manchester City didn't impress yet still cruised to victory against Fulham, Liverpool needed a little bit of Anfield magic to get them by an equally good Tottenham side, which means the two teams are still neck and neck for the Premier League title. LILYSILK 100% Mulberry Silk Terse Pillowcase Ivory Queen Standard King 19 Momme
As 2017 is about to come to an end, we thought it would be a good opportunity to look back at our favorite sexy ladies this year. The list includes … More


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A unique minimalist jewelry that has the power to bring back memories.
Minimalist Hornad river necklace is made in sterling silver with a custom designed silver "MiaBiel" closing.
A handtied thin black leather lace.
Necklace is shipped in a handmade black gift box with an elegant velvet pillow.
The widest part of the river is about 10 cm long
The lace or chain about 15 cm from the neck line long. Can be shorter on request.
Hornad minimalist necklace was designed to remind the nice moments of you or your loved ones. It flows through the second biggest Slovak town Kosice.
SKU: Hornad
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In progress
Bollard has been hit. Tarmac in footpath lifted and has created a trip hazard
Reported in the Safety Bollard - Damaged/Missing category anonymously at 13:01, Wed 13 November 2019
Sent to Northamptonshire County Council 4 minutes later. ref: 1844417.
Bollard has been hit and the tarmac that it is seated in has lifted. It's an area about 3' by 2' and has lifted 1" to 2". The bollard is upright but the lifted tarmac presents a trip hazard. It's adjacent to Hartwell village main through road and the footpath is used by children going to school. The location is outside 26 Park Road. The issue was first noticed on Friday 8th November.
Thank you for reporting a fault. We will use the information you have provided to assess and prioritise this fault in accordance with our published response times for fault reports.
State changed to: Investigating
Posted by Northamptonshire County Council at 13:05, Wed 13 November 2019
Thank you for your enquiry. The bollard has been made safe with temporary barriers and a works order has been raised and will be completed within 28 days to repair the bollard.
Posted by Northamptonshire County Council at 14:45, Fri 15 November 2019
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Main cast: Idina Menzel (Elsa), Kristen Bell (Anna), Jonathan Groff (Kristoff), Josh Gad (Olaf), Sterling K Brown (Mattias), Evan Rachel Wood (Iduna), Alfred Molina (Agnarr), Martha Plimpton (Yelana), Jason Ritter (Ryder), and Rachel Matthews (Honeymaren)
Directors: Chris Buck and Jennifer Lee
No, you're not going to be given a chance to let things go, because Elsa and Anna are back. Already, both genuine and knock-off merchandises of those two Disney princesses are clogging every shelf of everything here, and perhaps it is a blessing that no song in this sequel as instantly catchy as that song. You know which song. Mind you, the songs this time around are actually much better, but none is as immediately infectious as that song.
Frozen II is unnecessary, it's just a means to allow Disney to make even more money. There is really nothing here that constitutes must-see and must-know material, as this time around, Elsa and Anna plus their hangers-on the dude with the big nose, that annoying snowman, and that reindeer tag along on an adventure. Given that Elsa is this time around one of the cool kids rather than the underdog, she is immediately far less interesting. If anything, I'd argue that this is Anna's story, as it dwells quite significantly on her serious abandonment issues.
The movie opens with young Elsa and Anna listening to their father's story about he narrowly escaped with his life when he accompanied his father to a peace treaty with the Northuldra tribe who lives in the Enchanted Forest. The whole thing turns into a deadly fight, however, and when Agnarr came to, the spirits of the Enchanted Forest had sealed the whole place with a magical mist barrier to ensure that nobody will get in and cause trouble ever again. Then, their mother told the girls about the Ahtohallan, a magical singing river somewhere beyond the Enchanted Forest, where the waters contain memories and hence the answers to every question in the world.
Back in the present, Elsa begins hearing the Ahtohallan's song, and realizes that she's compelled to seek out that river and figure out what it wants with her. As she sings in Into the Unknown, she increasingly feels out of place among the good people of Arendelle, and a part of her believes that she can find a place where she truly belongs by seeking the river… but at the same time, she is afraid of jeopardizing the calm as well as her restored bond with her sister. The choice is taken from her, though, when she accidentally awakens the spirits of Enchanted Forest, which quickly cause enough mayhem in the kingdom and causing the people to evacuate the city. Since this is a cartoon, it makes perfect sense that both princesses seek out the Enchanted Forest, leaving no one with the authority to take over should both get eaten by a giant toad or something. What Elsa and Anna find will challenge everything they know about their family, their history, and their place in this world.
Now, let me put this out of the way first: I love this one far, far more than the movie that started it all. For one, scratch behind the surface, and it's a darker movie through and through. Anna is seriously crippled by the fear of losing her sister, while Elsa is still slowly suffocating from a sense of never belonging anywhere. Kristoff loves Anna, but he will have to grapple with the fact that she will always put her sister over him. Even Olaf, now a sentient and living creature with permafrost thanks to Elsa, is grappling with new concepts such as emotions.
The songs this time around are far more sweeping and even melancholic, although true to the Disney formula, anything dark is quickly swept up by inspiring determination to overcome the darkness. Anna's anthem The Next Right Thing is a gorgeous and striking example of this: it is a song for those who are spiritually broken. It is very easy to interpret the initial verses as Miss Perky Thang actually considering suicide as all seems lost to her. However, she soon, at first tentatively but later confidently, makes up her mind to keep going – one at a time, for now, with her prioritizing the next right thing to do as her next step to take. You know, as inspirational anthems go, this one isn't bad at all. No, it's glorious, and it's one that I feel may be a balm to young kids grappling with a dark place in their minds.
These songs narrate a big bulk of the story, and oh my, my heart is shredded as a result. Okay, story-wise, this one is pretty mediocre, with Elsa's abilities being a deus ex machina that solve every challenge thrown her way. The story itself is full of plot holes and relies quite a lot on lucky coincidences. However, and I'm still embarrassed to admit this, I have tears running down my cheeks during the last forty minutes or so of the whole thing. Through the songs, this movie does everything Terminator: Dark Fate and the recent Charlie's Angels boasted of wanting to do only to fail miserably: to put in the forefront plucky, strong female central characters and celebrate their sisterhood in ways that feel so natural, real, and cathartic. I was annoyed by Anna, Kristoff, and especially Olaf in the previous movie, but here, I cry for their ups and downs because I am so caught up in their trials and tribulations. I cry both happy tears and tears of heartbreak in this movie, mind you, and I never do that while watching a cartoon… well, other than the first The Land Before Time, that is, but please don't tell anyone.
If I do have a complaint, aside from the mediocre story, it's that the story brings up the sisters' inner demons, often in hauntingly poignant melodious ways, only to eventually sweep their issues aside for a hug and a happy ending. Yes, maybe it's too much to expect deep character arcs in such a cartoon, and perhaps it's enough that I have Show Yourself – the song in which Elsa gets an even more gorgeous gown and a rousing chorus that has me feeling all choked up inside. Even Olaf's song When I Am Older is quite dark – it's about a frightened person trying to rationalize the terrifying and often hurtful things in the world as things that will make sense and feel right when one is, well, older. In a way, it's can be interpreted as a terrified child's way of coping with abuse and worse, but hey, let's look at the cute things on the screen and tell ourselves that this is a Disney cartoon, so the song surely isn't that bleak. Right? But eventually, Olaf's inner demons are also swept aside for a hug and a happy ending. There are no permanent mental scars here, after all.
Frozen II isn't just so much darker and better than Frozen, it's essentially a very somber adult fantasy of abandonment issues, codependent attachment, and a few more things that will likely fly over the heads of younger children. Elsa and Anna are two damaged sisters who need one another to be a whole person. There are also all those songs, those feels, and that heady light-headed, warm rush in me as I stagger out of the cinema – stunned that this movie, of all things, has left such an impact on me. I'm not embarrassed by this, but please stop me should I begin entertaining terrible ideas like dressing up as Elsa.


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CS SoundStop combines all the benefits of our standard models with the added feature of measured acoustic ratings up to STC 43. This cavity slider is perfect for boardrooms or any other area requiring a level of privacy. Due to the weight of the door leaf it is recommended that the door does not slide fully back into the pocket.
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DEBATE: Domestic abusers don't have a profile, and there's no typical victim
In your marriage you feel appalled by yourself.Claudia Burger, Social Worker and Programme Director of ANEX and a domestic violence survivor
Mothers, sisters, daughters, friends - anyone of these women could be a victim of domestic abuse.
Domestic violence remains a pressing issue in the country. When looking at this issue from the outside, it is difficult to understand why women who are abused by their partners don't just leave. That is because we don't fully grasp the situation.
To help us understand this and to give some insights on the realities of domestic abuse, Pippa Hudson hosted a panel on domestic violence. Joining her were various women who work with victims of abuse in various spheres.
We often hear figures like one in six women or one in four are victims of abuse. Seeham Semaai, director at the Women's Legal Centre says the numbers are based on the women who actually report their abuse. She adds that women don't know about the acts that can protect them.
One of the challenges that we have are those are the actual reported ones. There are a lot of women who are still out there, who don't understand the purpose of the domestic violence act or the protection of harassment act that can assist them.Seeham Semaai, director at the Women's Legal Centre
Even if women are aware of these Acts, Semaai says the number of withdrawals of cases are high. This feeds into that perception that women who are abused could just leave - but often it is not that simple.
Nozuko Conjwa is a social worker at Nonceba Family Counselling and says that this could be attributed to the psychological abuse women in these situations face. She adds that many women feel like the can't leave an abusive relationship because the feel like they don't have support outside their relationships.
Another big factor, according to Nonceba, are cultural reasons that prohibit women from leaving a marriage. No matter haw bad the situation gets, women get told to stay with their abusers.
If you look at that, it is abuse on its own because the women we think are going to be able to assist us in terms of curbing domestic violence, are also perpetrating it by telling us to stay.Nozuko Conjwa, social worker at Nonceba Family Counselling
Claudia Burger is a social worker and programme director of Activists Networking against the Exploitation of Children (ANEX). She is also a survivor of domestic abuse. From her experience, women struggle to leave because they are fully stripped of all confidence.
Burger says one of the hardest things to deal with when in a violent relationship is the stigma attached to it. She says people will shame you for things you already feel ashamed about. When it comes to the actual abuse, you do feel appalled, yet you cling on to the good moments of the relationship - because you feel like you cannot leave.
The person who inflicts the violence is like a tsunami. You don't know when to expect what. I was abused in every way, beside physical. At 3 o'clock you start asking yourself, what do I need to prepare myself for tonight.Claudia Burger is a social worker and programme director of ANEX
When it comes to the abusers - there is no particular face or type of person. In fact, there is not such thing as a typical victim either. Pauline Perez, general manager at Nonceba Counselling Centre says women who come to them for help come from all type of backgrounds.
Burger adds that there is no way of telling who a perpetrator is. You have no way of saying only those people are abusers. In the case of her ex-husband, he was well liked in a crowd, and would be fine in a braai, but privately was very controlling and narcissistic.
Conjwa says that there are no profile for a perpetrator of abuse, but common issue that arises with them are anger, control and power.
The abusive partners come from different places. They're white, they're black, they're professionals, they're non-professionals - and so there is no such thing as a typical perpetrator.Pauline Perez, General Manager at Nonceba Counselling Centre
First for Women did a survey of survivors of domestic violence to find out more about how to help the situation. One of the biggest fears women have of leaving an abusive relationship is the economic factors. But the First for Women survey showed many survivors were able to find their feet quite quickly after leaving.
Robin Farrel, director at First For Women Insurance, says that 90 % of women found financial independence. It took them between three months and two years to find their feet, but they did it one way or another. Another key factor in overcoming abuse was great support systems, particularly of other women, who helped them regain their confidence and give them strength to leave.
Farrel says some of the best advice from survivors for those wanting to leave is to plan where they are going, keep evidence of abuse, keep a bad with all your important documents and be prepared to leave at any minute.
That's the message we want to give to those women who are currently victims. They can become survivors and they can become financially self sufficient.Robin Farrel, director at First For Women Insurance
Listen to the full panel discussion below:
This article first appeared on 702 : DEBATE: Domestic abusers don't have a profile, and there's no typical victim
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Amina Mohammed Poised To Become Un Deputy Sec-gen
SAN FRANCISCO, December 12, (THEWILL) – Nigeria's Minister of Environment, Amina Mohammed will soon be confirmed as a Deputy Secretary General of the United Nations.
If confirmed, NAN reports that she would deputise for newly Elected Antonio Guterres who is set to take office as the UN scribe on January 1, 2017 .
According to a tweet by Pamela Falk, CBS news reporter for the United Nations, the world body would soon release a statement confirming the appointment of the Buhari's Minister.
Until her appointment as a Minister of the Federal Republic of Nigeria in November 2015, she was the Assistant Secretary/Special Advisor to the United Nations Secretary General on Post-2015 Development Planning.
Amina Mohammed was also recently appointed by President Muhammadu Buhari to serve in the African Union (AU) Reform Steering Committee as the Nigerian representative.
The committee, headed by President Paul Kagame of Rwanda is tasked with ongoing institutional reforms of the AU Commission and its organs.
The committee which comprises eminent persons from the continent will work on part-time basis to produce a report for presentation to the 28th African Union Summit in January 2017.
A versatile and accomplished development practitioner in the public, private and civil society sectors with over three decades experience, Mrs Mohammed had served as the Senior Special Assistant to the President of Nigeria on MDGs, serving three Presidents over a period of six years.
THEWILL recalls that the new Secretary General took his oath of office on Monday .
Story by Oputah David


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With SecurOS, ISS offers security professionals around the world an all-encompassing and open platform for video surveillance. Barco, for its part, develops flexible and mission-critical visualization software and hardware solutions including the CMS; an integration-friendly, advanced software suite for easy video wall management, inter-control room collaboration and swift decision-making based on the sharing of visual information. While both systems have proven their strengths with various customers and diverse situations over the past six years, this software integration now becomes a native feature of SecurOS.
"In Barco we have found the perfect partner to extend the reach and the potential of our product offerings and strengths", says Aluisio Figueiredo, COO at ISS. "Our alliance enables a seamless integration of our products, expertise and experience. It will create greater flexibility and maximize the value of our solutions for our joint customer community. In short, our customers will enjoy more options and faster response capabilities without needing additional training."
Maximized interoperability
Barco-powered video walls can now be controlled either with the Barco CMS Sidebar, through an ISS SecurOS client, or using a mix of both, based on the preferred workflow in customers' control centers. Through the SecurOS Barco control panel, control room operators can select pre-configured CMS perspectives – sets of visual sources providing situational awareness. Using the SecurOS user interface, operators can easily drag and drop any video source onto the active perspective on the Barco video wall, as well as switch between perspectives on the fly.
The software integration offers control room operators greater situational awareness and shorter reaction times thanks to the drag-and-drop functionality. What's more, since all of these functionalities will be offered within a familiar environment, they will require little to no additional training.
A strategic alliance for the benefit of customers worldwide
The Barco/ISS alliance will enable a reach spanning the entire globe, as the integrated system will be available to control room professionals around the world. Sergio Martinez, Barco Country Director Mexico, underpins the importance of this new partnership to Barco: "Through ISS, Barco can tap into a vast experience and knowhow of local markets and applications, both in Mexico and in the whole of Latin America, and far beyond these borders. Close collaboration between our companies will pave the way for new projects and solutions tailored to evolving needs of our end-customers."
With its Strategic Alliance Partner program – www.barco.com/alliances – Barco looks to join forces with software developers, reference design partners and manufacturers around the world. Its goal is to create a win-win situation for all parties involved. Suchit Rout, Director – Global Strategic Alliances at Barco: "Partnerships like the one with ISS allow us to offer the best joint solutions on the market which are already pre-validated. Drawing on complementary strengths, we look to provide integrated solutions to the benefit of our common customers. They profit from simplified deployment, optimized efficiency and reduced risks."
Discover ISS and Barco at Expo Seguridad
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2 area students receive MSSU Dolance Awards
Leadership recognition to Brewer, Burns
Monett and Pierce City students at Missouri Southern State University in Joplin received honors during the 27th annual Glenn D. Dolence Leadership/Service Recognition Assembly, held April 4 at Missouri Southern State University in Joplin.
Emily Brewer, a secondary education major from Monett, and Ashley Burns, a biology, ecology and conservation student from Pierce City, were among the 35 students recognized this year during the awards ceremony. Student Services and Omicron Delta Kappa at Missouri Southern sponsor the awards, which are presented to graduating students who have demonstrated outstanding leadership skills on campus and who have made a difference in the areas of campus organizations, student employment, student services or athletics.
The awards are named in honor of Dr. Glenn D. Dolence, former vice president for Student Services at Missouri Southern, who retired from MSSU in 1996 after 27 years of service. He later served as mayor of Webb City, Mo., and has held a variety of other posts in the community.


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The first conference of The Taureaux du Panthéon was held on March 21st 2017. Here's a summary and photos of this enriching night.
Speaker 1: Julien LAUMAIN
-compliance officer
-BNP Paribas
Compliance officer :
-limit the risks of the bank
-reputational risk: keep a good image
-risk of fines/penalties
-risk of loss of approval: especially in the USA
-basic operational risks
-to focus on 5 trades
-customer knowledge
-financial security: fight against money laundering, terrorism
-the question of market abuse
-preservation of the client's interest
-top manager= decision
-close to the operational
Roles ;
—identify the rules of the job and the risks
—evaluate the risks (with and without devices)
—check-up(front/back/middle office)
—avoid making money on risks
—the aspect of advice : on the regulation part, participation to all projects, macroeconomic vision of the projects
—train newcomers : on the deontology, anti money laundering, sensibility, evolving regulations…
—to learn : learn about the regulation and to be alert about these new regulations and their consequences
—correspondence with regulators (ex : AMF)
— cartography of risks on the whole group
—intervention within teams on all the tools at their disposal.
— database
—define roles and therefore the devices
—finally a check-up on all the projects containing risks
—consultation of projects and datas
Speaker 2: SABRINE AOUIDA
— analyst
AMF: Authority that exists to accompany companies and not to only sanction them. Stock companies, make the Parisian financial place as competitive as possible.
Issues in all bank services and management companies :
—the internal control's culture
—need to know the regulatory isuues
—texts to know, directives..
S = question for Sabrine
J= question for Julien
deux = question both
S: a day:
— different everyday
— in contact with the thirds of AMF
— Actual missions: a lot of pedagogy, learning of risks and instruments, accompanying savers, detect the abusive and disloyal marketing practices. In relation with the public (future management companies, cabinets, pro..) that have questions on the AMF doctrines.
— depends on actual situation
— we are formed daily on new themes
both : perspective on career evolution
—rise in rank
—rise in the group
—go abroad
—change jobs
—depends on what we want
—secretarial position in banks
— join a regulator
—change directions at AMF
—accomapny companies in the product approvals that they manage
—no changes to predict
—move into management companies
—control and manage risks
J: compliance pole, help to bypass regulations?
— power of veto
— huge culture of risks
—level control, general inspection, AMF
—criminal liability
both: application of directives:
—if there are hesitations we ask a regulator
—the regulator will have the last word on how to apply a point on a directive
Moderators : Hugo Daugeron-Mijoin & Rouzbeh Moshfeghi
A very big thank you to Ryan Ould Hocine for organizing this conference and to our two speakers.


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Police seek to identify man accused of stealing beer from Weis Markets
YORK COUNTY, Pa.– Police are seeking to identify a man accused of stealing beer from a grocery store.
Authorities are attempting to identify the man in the above photo in connection to the theft of beer from the Weis Markets store in the 1800 block of Roosevelt Avenue in West Manchester Township on August 12.
If you have any information regarding the identity of the suspect, you're asked to contact police at 717-792-9514.
39.947835 -76.791356


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The IFTAs are upon us again. The great and good, the glamorous and glorious, the gorgeous, gregarious and gratuitous are poised on the red carpet, ready to be piped into our living rooms. Let's celebrate the best of the best in Irish film and television.
However, a quick tot of the named nominees this year shows that only 27% have a double X chromosome – that's 40 women across 41 categories. Well, across 34 categories actually, as there is zero female representation in Direction, Script*, Cinematography, Production Design, Animation, Score and Sound. Creative talent is not the sole preserve of a single gender, and I quote the great thinker Lupe Fiasco when I ask, 'Where my ladies at?'
Lupe got me to pondering – maybe very few women work in the film and television industry, and that's why the nominations aren't flooding in. First stop IBEC. Unfortunately, for my purposes, IBEC has not asked for an employee gender breakdown since 2006, which would have looked at 2005 productions. However, it is interesting to note that these 2005 numbers, skewing 57% male to 43% female are not a million miles away from the current EU figures on general Irish employment: 54% male, 46% female.
A quick ring around the various colleges and universities offering film, media and communications courses yields more interesting results. Where high Leaving Cert points are required, and on the more arts-centred courses, the student split was close to 50/50, skewing female. On the more technical courses, the split widened, skewing 70/30 male and in some cases reaching as far as a 90/10 male/female divide.
To examine female representation in the creative origin of film projects, I took a quick scan of the Irish Film Board's 2012 development slate. It reads depressingly. Of the successful directors, only 28% are female, and of the writers, a paltry 25%. There are no figures available on the gender split of applicants, so we cannot surmise whether these director and writer figures represent a success rate of 10%, 50% or 100% of applications from female creatives.
The IFTA Irish Film Board Rising Star awards, anticipated to be representative, one imagines, of the future of Irish talent, acknowledged female skill and ability with only 1 out of 5 nominees. Across the pond, where BAFTA are allowing the public to vote for their Rising Star category, we see, from 5 talented young things, that 4 are women.
From this, albeit not entirely scientific statistical analysis, we can make some conclusions. 1. Not enough women are being encouraged into the technical end of filmmaking. 2. As an industry, film and television is not abnormally more pro-male than other Irish industries. 3. There is a clear dearth of female voices and stories being developed, produced, making it to market and ultimately to a shiny podium in the Convention Centre.
Which brings me back to the IFTA nominations. What better way to encourage young women into an industry than for them to see talented female creatives recognised, lauded and championed? How about a bit of role model action for the girls at the back? Anyone…?
2011 was a particularly sad year for female thespians – apparently there weren't eight actresses of sufficient merit to be found in this country, so IFTA created a combined Lead Actress in a Film/TV Role category. I'm not crying conspiracy – clearly enough quality actresses weren't put forward for selection.
So who is responsible for the selections? In a tidy piece of circularity, we turn again to our industry – it is Irish producers, production and post-production houses that nominate their colleagues and work. IFTA members then vote as to who goes through for the nominee shortlist, and the chance to strut their stuff on the red carpet. When I contacted IFTA to find out their membership gender split, they did not have exact figures but assured me it's about 50/50.
This, I must admit, leaves me in a bit of a bind as I come to a conclusion. Some small part of me had hoped my fleeting foray into the world of investigative journalism would yield Pulitzer results, and uncover a Machiavellian misogynistic machine working devilishly behind the scenes, thwarting women from claiming their golden statues… In reality, there are no such easy, flippant answers.
Ultimately, audiences and industry alike are being cheated. We need a variety of voices, stories and experiences on our screens in order to build and maintain a diverse, exciting and successful industry and culture. We need to encourage more young women to make their voices heard. We need to champion and, crucially, support women already excelling in their chosen field**. Our industry's future depends upon it.
If you're interested in finding solutions to the under representation of women in the Irish film and TV industry, please add your voice on 28th February – see Underground Films' Facebook page for more details.
* 'Jump' is based on the stage play 'Jump' by Lisa McGee, though she herself is not nominated for the screenplay.
** Much as I would like to segue into a discussion on maternity leave for self-employed women, long working hours and childcare, printing space decrees I must desist.
Rachel Lysaght
Rachel Lysaght is an award-winning Film & TV Producer, and a graduate of the European EAVE programme and the Samuel Beckett School of Drama in Trinity College, Dublin, Ireland. She is a lecturer for Filmbase Dublin's Masters in Creative Documentary Production from Staffordshire University, UK.
Recent producing credits include new release THE RUNNER (Director Saeed Taji Farouky 2012), THE RELUCTANT REVOLUTIONARY (Director Sean McAllister 2012), DREAMS OF A LIFE (Director Carol Morley 2011) and THE PIPE (Director Risteard Ó Domhnaill 2010).
This article first appeared in issue 144 Spring 2013, the last ever printed issue of Film Ireland, which was published 14th February 2013.


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"¡El Inferno Llega a Casa!": The synopsis for this issue has not yet been written.
- No... paybacks are.
- — Lono
Appearing in "¡El Inferno Llega a Casa!"
Featured Characters:
Supporting Characters:
- Father Manny (priest)
- Sister June (nun)
- Cortez (gangster)
- Craneo (Dies)
- Maddon (gangster)
Other Characters:
- Cesar (Dies)
- Paulo / Pico (Dies)
- Mexico
- Durango (city)
- San Sebastian Orphanage
Synopsis for "¡El Inferno Llega a Casa!"
- Synopsis not yet written.
- This book was first published on December 8, 2013.
- No special notes.
- No trivia.
See Also
- Write your own review of this comic!
- Discuss 100 Bullets: Brother Lono Vol 1 7 on the forums
- Cover gallery for the 100 Bullets: Brother Lono series
- Images from 100 Bullets: Brother Lono Vol 1 7
Recommended Reading
Links and References
Summary Needed
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