text:
The Lumberjacks will look to remain at the top of the Southland Conference standings, while the Ladyjacks will try to overtake second place with both teams playing in a non-conference midweek game followed by a weekend conference series.
The Ladyjacks are coming off a series sweep of the Houston Baptist Huskies last weekend, and they currently sit third in the SLC standings with a 6-3 conference record— one game behind second- place Southeastern Louisiana.
SFA will play Prairie View A&M at 3 p.m. Wednesday at the Lady Panther Softball Stadium to open this week's slate of games. PVAMU (18-7; 6-0) is perfect in SWAC play and coming off two-straight sweeps of fellow SWAC counterparts Alcorn State and Jackson State.
PVAMU boasts three players—Diavionne Dewalt (.421), Destinee Williams (.378) and Laurisa Hernandez (.371). Hernandez leads the team in RBIs with 20 followed closely by Williams with 19. The pair also lead the team in home runs with four each.
The Panthers have two pitchers who have sub-three ERAs. Crystal Castillo boasts a 2.20 ERA, a 1.09 WHIP an 8-3 record. Following closely is Ashley Higgins with a 2.68 ERA and a 1.38 WHIP.
Following the PVAMU matchup, SFA will travel to Corpus Christi to take on the Islanders of Texas A&M- Corpus Christi in a weekend three-game series.
TAMUCC ranks 11th in the SLC standings, with a 4-27 overall record and a 1-8 conference record. Their lone conference win came against Houston Baptist on March 8.
The Ladyjacks will play a double header at 1 and 3 p.m. on Friday, followed by the final game in the series at noon Saturday.
SFA is coming off its first series loss to an SLC opponent this season after losing 2-of-3 games last weekend to the Central Arkansas Bears. However, the Jacks still sit atop the SLC standings with a conference record of 7-2.
The Lumberjacks will battle the University of Houston at 2 p.m. Wednesday at Jaycees Field. UH is currently 11-11 on the season and 1-2 in conference after dropping the series to UConn last weekend.
The Cougars are led by Joe Davis who boasts a .333 BA, leading the way in RBIs with 27 and eight home runs. Davis also has a .988 fielding percentage. Following Davis was Jared Triolo with a .267 BA and has shown plate discipline by drawing a team-leading 13 walks.
Houston is led from the mound by Fred Villarreal who has a 1.05 ERA and a perfect 2-0 record. Following closely is Nolan
Bond who has a 2.66 ERA. After the Houston matchup, the Lumberjacks will take on SELA in a three-game series over the
weekend at Jaycees field. The Lions are coming off a series sweep over the Abilene Christian Wildcats. With the sweep of the Wildcats, the Lions have an overall record of 11-12 and a conference record of 4-2 and sit fifth in the SLC standings.
SELA are led by Kyle Schimpf who has a .318 BA, 20 RBIs and two home runs. Cody Grosse follows closely behind with a .309 BA 19 RBIs. Nick Ray leads the team with RBIs and ranks in the top five in the conference with 26.
Corey Gaconi leads the team from the mound with an ERA of 2.79 and a WHIP of 1.07. Gaconi has an overall record of 2-2 and 36 strike outs to add to his tally.
The Jacks will start off the series with SELA at 6:30 p.m. Friday,followed by Saturday's game at 2 p.m. and Sunday's matchup at 1 p.m.


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Critical fluctuations in domain-forming lipid mixtures
Edited by Harden M. McConnell, Stanford University, Stanford, CA, and approved September 4, 2007 (received for review April 16, 2007)
Critical fluctuations are investigated in lipid membranes near miscibility critical points in bilayers composed of dioleoylphosphatidylcholine, chain perdeuterated dipalmitoylphosphatidylcholine, and cholesterol. Phase boundaries are mapped over the temperature range from 10°C to 60°C by deuterium NMR. Tie-lines and three-phase triangles are evaluated across two-phase and three-phase regions, respectively. In addition, a line of miscibility critical points is identified. NMR resonances are broadened in the vicinity of critical points, and broadening is attributed to increased transverse relaxation rates arising from modulation of chain order with correlation times on a microsecond time scale. We conclude that spectral broadening arises from composition fluctuations in the membrane plane with dimensions of <50 nm and speculate that similar fluctuations are commonly found in cholesterol-containing membranes.
The first direct observation of immiscible liquid phases in model bilayer membranes was presented only recently, for membranes containing three or more components (1, 2). However, by the early 1970s, it was already clear that lipids in membranes with elevated cholesterol have high chain order yet diffuse as in a liquid (3–6). This liquid-ordered (Lo) phase of lipids (7) is distinguished from the familiar liquid-disordered phase (Lα or Ld) found in bilayers of pure lipids above the chain melting temperature (3–14). Pure lipids below the chain melting temperature are in the solid or "gel" phase (So). Coexisting liquid-ordered and liquid-disordered phases are easily visualized in giant unilamellar vesicles by fluorescence microscopy when membranes contain cholesterol and two types of lipids: one with a high gel–fluid transition temperature and one with a low gel–fluid transition temperature (15–21). Coexisting liquid phases are also easily visualized in membranes of plasma membrane vesicles containing a wide spectrum of lipid and protein components (22).
The research presented here represents a culmination of several years' work in our laboratories on ternary membranes of dioleoylphosphatidylcholine (DOPC), chain perdeuterated dipalmitoylphosphatidylcholine (DPPC-d 62), and cholesterol (Chol) (15–18, 23, 24). It has resulted in a straightforward thermodynamic description of micrometer-sized coexisting phases and of submicrometer composition fluctuations in membranes containing cholesterol. Here we use deuterium (2H) NMR to assemble the DOPC/DPPC-d 62/Chol phase diagram. NMR allows us to directly measure differences in ordering and partitioning of DPPC-d 62 between coexisting phases. In addition, NMR produces results free from artifacts associated with fluorescent probes (24) or oxidation of unsaturated lipid chains (15, 25). We use our NMR data to identify quantitative miscibility phase boundaries and to solve for a large array of tie-lines. Tie-line endpoints yield quantitative compositions of the two liquid phases and provide independent verification of the miscibility phase boundary. Furthermore, we unambiguously determine the extent of a three-phase So-Ld-Lo coexistence region, a task that is difficult to accomplish by other methods (16, 17).
In the theory of critical phenomena, lateral inhomogeneities arise because the energy required to maintain regions of differing composition becomes vanishingly small, and thermal motions lead to composition fluctuations over a wide range of time and distance scales (26, 27). The size of the largest probable fluctuation is described in terms of a correlation length, which diverges as the critical point is approached. We find that broadening of resonance peaks occurs for membranes near known critical points. We attribute broadening to composition fluctuations on length scales of <50 nm in these membranes.
Results and Discussion
2H NMR spectra from multilamellar vesicles of 31 distinct compositions of DOPC, DPPC-d 62, and Chol for temperatures between 5°C and 60°C were acquired, and representative spectra from lipids in uniform liquid phases are shown in Fig. 1 a and b. Each spectrum is composed of superimposed resonances of symmetric 2H quadrupole splittings whose magnitudes reflect the time averaged orientations of 2H–C bonds with respect to the bilayer normal. Here, 2H–C bonds occur within both hydrocarbon chains of perdeuterated DPPC-d 62 lipids. The narrowest and most prominent splitting is from the terminal methyl groups, whereas wider splittings arise from superimposed resonances of methylene groups along the chains (23).
2H NMR is well suited for resolving whether lipids are in single phases (Ld, Lo, and So), in two coexisting phases (Ld-Lo, Lo-So, or Ld-So), or in all three phases simultaneously (Ld-Lo-So) (18, 23). DPPC-d 62 chains in the Ld phase have low chain order parameters and produce a series of narrow and well resolved splittings as in Fig. 1 a. In contrast, lipid chains in the Lo phase have significantly higher order and produce wider splittings (Fig. 1 b). Ld- and Lo-phase spectra are also distinguished by the number of quadrupolar splittings from terminal methyl groups. A single terminal methyl group splitting is found from DPPC-d 62 in the Ld phase, whereas two distinct methyl splittings are found from DPPC-d 62 in the Lo phase (13, 23). Spectra from lipids in the So phase are distinguished by their larger quadrupolar splittings and by their poorly resolved resonances. These defining features apply generally to single-phase spectra over a wide range of experimental conditions. The detailed appearance of each Ld, Lo, and So spectrum depends strongly on temperature and composition. For vesicles containing coexisting phases, the relative intensities of the single-phase (Ld, Lo, and/or So) contributions superimposed in each spectrum provide direct measurements of DPPC-d 62 partitioning into the different phases (Fig. 1 d and e).
DPPC-d 62 spectra like those in Fig. 1 are used to quantitatively calculate tie-lines, with errors, for a wide range of temperatures and compositions as described in Materials and Methods [Fig. 2 a–d and supporting information (SI) Table 1]. Tie-lines span all binary coexistence regions in the DOPC/DPPC-d 62/Chol ternary phase diagram. Knowing the location of tie-lines within the Ld-Lo coexistence region is valuable because their endpoints can be read off to give the compositions of the Ld and Lo phases. Tie-line endpoints also provide an independent measurement of the phase boundary. When the compositions of two vesicles lie along the same tie-line, the vesicle membranes contain different ratios of the same two coexisting phases in proportions given by the Lever rule (16). Here, we solve for tie-line slopes by searching the composition space for spectra that superimpose different ratios of the same two endpoint spectra, indicating that they contain the same two coexisting phases. The location of tie-line endpoints is then determined by inserting the measured DPPC-d 62 partition coefficients into the Lever rule.
DPPC-d 62 spectra are also used to determine all membrane compositions for which three distinct phases coexist in the membrane. Within a Gibbs phase plot as in Fig. 3 a, the three-phase region fills a triangle. Within this triangle, all acquired spectra are a linear superposition of the same three Ld, Lo, and So endpoint spectra. This property is exploited to evaluate the three triangle vertices (values given in SI Table 2). In all cases, calculated tie-line and three-phase endpoints agree with phase boundaries determined through visual inspection of acquired spectra. Miscibility transition temperatures are recorded in SI Table 3. The boundaries of two- and three-phase regions at different temperatures are assembled into the three-dimensional phase diagram shown in Fig. 3 b.
Compared with our earlier work (15, 23), the current phase diagram contains the same phases, but the boundaries between phases lie in slightly different positions because here we use exclusively NMR (24). In addition, we have evaluated tie-lines through a large number of compositions using an entirely quantitative method, and the errors in tie-line slopes and endpoints are significantly reduced. Previously, we were not able to determine Ld-So or Lo-So tie-lines, or to unambiguously locate the vertices of the three-phase region. These advances lay the groundwork for our investigation of critical point behavior described below. For example, knowing the location of tie-lines in the Ld-Lo coexistence region makes it possible to identify the compositions of miscibility critical points. One critical point is at the composition with the highest miscibility transition temperature. At lower temperatures, critical points fall near tie-lines located at the high cholesterol edge of the miscibility phase boundary, as illustrated in Fig. 3 b.
We observe significant spectral broadening for membrane compositions near known miscibility critical points. This is seen as loss in resolution in the acquired spectra. Examples are shown in Fig. 1 c and i, which show poorly resolved terminal methyl splittings flanked by featureless shoulders that slope gradually to low intensity. This broadening is quantified by fitting the methyl peaks of dePaked spectra (28) with Lorentzian lines to extract peak widths (Fig. 2 e–h). Maximum broadening occurs near known critical points. Fig. 2 g and h shows that broadening is still detectable up to ≈5°C above the highest critical temperature, which lies between 27.5°C and 30.0°C for 1:2 DOPC/DPPC-d 62 plus 20% Chol.
To investigate the origin of spectral broadening near miscibility critical points, we conducted a series of relaxation measurements (Fig. 4), focusing on the composition 1:2 DOPC/DPPC-d 62 plus 20% Chol because its spectra exhibit maximal broadening (Fig. 1 i). Experiments were conducted to measure the longitudinal (R 1) and transverse (R 2) relaxation rates of 2H nuclei of DPPC-d 62 both in the presence and absence of magic angle spinning (MAS), and of 1H nuclei of DOPC with MAS (Fig. 4 a–e). In all cases, R 1 smoothly increases as temperature decreases. In contrast, R 2 rises sharply as the critical point is approached from higher temperature and then rapidly falls as liposomes enter the two-phase region.
Values of R 2 set a lower limit on the width of resonances, Δν1/2, according to Δν1/2 = R 2/π, when a Lorentzian shape assumed. The measured peak widths are in excellent agreement with values calculated from R 2 relaxation rates (Fig. 4 e and f), indicating that spectra are, to a good approximation, homogeneously broadened. Contributions to broadening due to sample inhomogeneity and exchange effects appear to be insignificant. It is interesting to note that both DOPC and DPPC resonances are broadened. Although it is not often discussed, order parameters of both DOPC and DPPC lipid chains are composition-dependent (29), and our results show that both DOPC and DPPC participate in fluctuations near critical points.
Following the analysis below, we find that transverse relaxation rates, R 2, must be enhanced in membranes near critical points as a result of modulations of chain order that occur with correlation times between 0.1 μs and 10 μs. Spin relaxation arises from modification of quadrupolar interactions (2H NMR) or dipolar interactions (1H NMR) by lipid dynamics that are described by spectral density functions J(2ω), J(ω), and J(0), where ω = 2πν, and ν is the NMR resonance frequency. Longitudinal relaxation rates, R 1, depend solely on J(2ω) and J(ω). These spectral densities arise from fast motions with correlation times from picoseconds to nanoseconds, such as isomerization of lipid hydrocarbon chains. Transverse relaxation rates, R 2, depend on J(2ω), J(ω), and also J(0) (30). Contributions to relaxation from J(0) grow steadily with increasing correlation time. Significant contributions appear at ≈0.1 μs and continue to grow until spectra from different molecular conformations and environments convert from the rapid exchange limit to signal superposition, which occurs at ≈10 μs for 2H NMR and ≈100 μs for 1H NMR. The data in Fig. 4 indicate that transverse relaxation rates (R 2) display an anomaly near the critical point, but longitudinal relaxation rates (R 1) do not. Changes in J(2ω) or J(ω) would impact both R 1 and R 2; therefore, only those motions that affect J(0) are responsible for observed increases in R 2 near critical points. We conclude that motions that alter lipid order with correlation times from 0.1 to 10 μs are greatly enhanced in the vicinity of critical points.
What might constitute these motions? Time scales can be converted to distances if normal two-dimensional diffusion is assumed, 〈x 2〉 = 4Dτ, where D is the lipid lateral diffusion constant [≈10−11 m2/s (31)]. On a time scale of 0.1–10 μs, lipids traverse distances of 2–20 nm. Therefore broadening could arise from lipid diffusion between very small fluctuations with long lifetimes. Radhakrishnan and McConnell (32) have recently shown that the dynamic behavior of complexes near a critical point may enhance R 2 rates sufficiently to explain experimental observations (32).
Fluctuations on larger time and distance scales have been described in a similar system investigated by fluorescence microscopy (18). In the current experiments, R 2 rates alone explain all spectral broadening; therefore, we observe no evidence of additional inhomogeneous broadening related to large fluctuations. This outcome is not surprising if the length scale of critical fluctuations (ξ) scales as in a two-dimensional Ising model, ξ ≈ f (+)ξo T c/(T − T c), where T c is the critical temperature (≈300 K), f (+) is an amplitude factor for T > T c (≈0.5), and ξo is on the order of a lipid diameter (≈1 nm) (27, 33). In this model, inhomogeneous contributions to broadening are only expected for temperatures and compositions very close to the critical point (e.g., at the critical composition, ξ ∼ 50 nm for T − T c ∼ 3°C). The amplitude factor is approximately three times smaller for T < T c compared with T > T c [f (−) ∼ 0.17] (33), providing a plausible explanation for why R 2 values in Fig. 4 decrease faster for T < T c than for T > T c.
There is a large body of experimental work suggesting submicrometer organization in model membranes containing cholesterol (9, 13, 34, 35). For example, we (23) and others (13, 36) have previously interpreted broadened 2H NMR spectra as evidence for submicrometer phase-separated domains. Our current results suggest an alternate interpretation, that broadened 2H NMR spectra instead indicate the presence of fluctuations within a single liquid phase in membranes near a miscibility critical point. Similarly, we can imagine that fluctuations provide a mechanism by which lipids and proteins organize on submicrometer dimensions in cholesterol-rich biomembranes in the vicinity of a miscibility critical point.
Materials and Methods
All lipids were acquired from Avanti Polar Lipids, stored at −20°C, and used without further purification. Multilamellar vesicles were prepared as described previously with either 2.5 or 5 mg of deuterated lipid (18). NMR experiments were conducted on a Bruker DMX500 spectrometer operating at a 2H resonance frequency of 76.8 MHz. 2H NMR spectra were acquired from 60°C to 5°C at increments of 2.5°C with a quadrupolar echo sequence and processed as described in ref. 23. DePaked spectra were calculated by using the iterative method described in ref. 28. The precise lipid compositions and lipid/cholesterol ratios were determined by dissolving samples in chloroform and conducting high-resolution 1H NMR experiments. 1H MAS NMR experiments were conducted at a 1H resonance frequency of 500.1 MHz and a spinning frequency of 10 kHz. The sample temperature in the spinning rotor was calibrated by monitoring the gel–fluid transition of fully hydrated DPPC-d 62 and DMPC-d54 bilayers.
Relaxation experiments were conducted by using multilamellar bilayers composed of 1:2 (mol/mol) DOPC/DPPC-d 62 plus 20% Chol over the temperature range from 50°C to 12°C at increments of 2°C. The 2H transverse relaxation rates (R 2) without MAS were measured with the quadrupolar spin echo sequence (π/2-τ-π/2-acq) by incrementing τ (37). The reduction of signal intensity in the methylene region of DPPC-d 62 corresponding to methylene segments near the lipid glycerol group (the so-called order parameter plateau) was fitted to a single exponential decay with increasing τ. Longitudinal relaxation rates, R 1, were measured with an inversion-recovery pulse sequence followed by signal detection with a quadrupolar echo (π-τ-π/2–50 μs-π/2-acq). R 2 and R 1 of DOPC in 1H MAS NMR were measured by a Hahn echo and an inversion-recovery pulse sequence, respectively. The peak at 2.1 ppm was used for the DOPC relaxation measurements and arises from the methylene groups adjacent to the double bonds of oleic acid.
Tie-Line Determination.
Spectra acquired from membrane compositions along a specific tie-line are a linear superposition of spectra from single-phase membranes with compositions corresponding to tie-line endpoints. For example, if a spectrum S(φ⃗, ν) is acquired at a composition φ⃗ = [%DOPC, %DPPC, %Chol] within the Ld-Lo coexistence region, where S(φ⃗Ld, ν) and S(φ⃗Lo, ν) are spectra acquired at compositions φ⃗Ld and φ⃗Lo corresponding to the tie-line endpoints, ν is spectral frequency, and A is the fractional partition coefficient of DPPC-d 62 into the Ld phase. For this analysis, all spectra were normalized to constant integral intensity. A large number of spectra were acquired from membranes with compositions within the two-phase region. When two acquired spectra have compositions φ⃗1 and φ⃗2 that lie along the same tie-line, endpoint spectra and partition coefficients can be determined by using spectral subtraction according to (13) where the constants B and C describe the linear combination of the acquired spectra S(φ⃗1, ν) and S(φ⃗2, ν) needed to produce a pure Ld or Lo endpoint spectrum. The constants B and C are algebraically related to partition coefficients and can be used in conjunction with the Lever rule to evaluate the composition of membranes at tie-line endpoints.
This technique has been applied to various two-component lipid mixtures (13, 36, 38), where thermodynamic rules stipulate that all compositions within the two-phase region lie along the same tie-line at constant temperature (16). In these earlier studies, subtraction coefficients were determined through visual inspection of subtracted spectra. Here, the constants B and C are determined quantitatively by performing a least-squares minimization against a single-phase reference spectrum acquired at a composition close to the desired tie-line endpoint. Because reference spectra are not necessarily acquired exactly at the tie-line endpoint compositions, we introduce an additional fitting parameter that stretches or contracts the frequency axis of the reference spectrum to compensate for minor temperature- and composition-dependent changes in these spectra. This operation assumes that order uniformly increases or decreases for all C–2H bonds along the chain and is valid for modest changes in temperature and composition (18, 39).
Evaluating tie-lines in three-component mixtures is complicated by the fact that the tie-line slope is not constrained (16), and it cannot be assumed that any two given compositions will lie along the same tie-line. This is addressed by recognizing that two compositions lie along the same tie-line only if it is possible to completely subtract the Lo-phase spectra without significant distortion to the Ld component. One composition is chosen to lie along a given tie-line, while a second composition along the same tie-line is determined by searching for a second spectrum in the composition space that can be linearly combined with the first spectrum to resemble an Ld-phase reference spectrum. Limitations from the finite sampling of compositions within the two-phase region are overcome by interpolating between spectra acquired from membranes with fixed DOPC:DPPC-d 62 ratios. We quantitatively determine tie-line slope and error bounds by performing a least-squares minimization between the calculated Ld endpoint spectra and an acquired Ld reference spectrum using three fitting parameters: the cholesterol composition of the interpolated spectrum, the linear combination of the two spectra needed to obtain the reference spectra, and the factor by which the frequency axis of the Ld reference spectra was scaled.
Once the tie-line slope is known, the standard method of spectral subtraction described above can be used to determine the location of tie-line endpoints. In a second fitting step, we obtain the remaining parameters needed to quantitatively determine the tie-line endpoints, and errors, by performing a least-squares minimization against a single-phase Lo reference spectrum. An example demonstrating the sensitivity of this method is shown in SI Fig. 5.
Three-Phase Triangle.
Within a three-phase region, each acquired spectrum is a linear superposition of three endpoint spectra corresponding to DPPC-d 62 in Ld, Lo, and So phases: formula φ⃗Ld, φ⃗Lo, and φ⃗So are compositions at the triangle vertices and all spectra are normalized to constant integral intensity. Mathematically, this is equivalent to stating that each value of S(φ⃗, νi) at a given frequency νi lies on a plane defined by the points S(φ⃗Ld, νi), S(φ⃗Lo, νi), and S(φ⃗So, νi), and that the position of S(φ⃗, νi) on this plane relative to the endpoints is parameterized by the constants D and E.
Because the endpoint spectra and compositions are unknown, this same plane is defined by using three (or more) experimental spectra acquired from compositions within the three-phase region by solving a system of linear equations: where φ⃗j are the compositions of the acquired spectra, and φ⃗DOPC, φ⃗DPPC, and φ⃗Chol are compositions of the three single-component membranes. Spectra for three separate compositions are needed to evaluate three basis spectra S(φ⃗DOPC, νi), S(φ⃗DPPC, νi), and S(φ⃗Chol, νi). When spectra for more than three compositions are available at a given temperature, a best-fit plane is calculated by using least-squares minimization. This procedure defines the linear coefficients and basis spectra that are combined to reproduce spectra acquired within the three-phase region. It is worth noting that the three calculated basis spectra are not themselves physical because they correspond to single-component compositions located outside of the three-phase triangle, in a region where the linear relationship between acquired spectra no longer applies. Each vertex of the three-phase triangle is determined by finding the linear combination of basis spectra needed to produce a pure Ld, Lo, or So endpoint spectrum. As in the tie-line analysis, this is accomplished by performing a least-squares minimization against a reference spectrum acquired at a composition near the suspected triangle vertex. In some cases, calculated endpoint spectra from adjacent binary tie-lines are used as reference spectra instead of raw experimental data. The effectiveness of this method is demonstrated in SI Fig. 6.
The relative intensities of endpoint spectra needed to produce spectra acquired within the triangle (denoted by the constants D and E above) are given by the three-phase Lever rule. In practice, the relative intensities of superimposed spectra can be altered by the quadrupolar echo sequence. This is particularly relevant when evaluating the intensity of So-phase spectra (36). When this attenuation is taken into account, we find that acquired spectra are well represented by weighting the Ld, Lo, and So endpoint spectra within the Lever rule (SI Fig. 6e ).
Evaluation of Errors.
In all cases, errors in fitting parameters are determined directly from the covariance matrix at the solution of the least-squares minimization. The diagonal elements of the covariance matrix are the squared standard errors of the fitting parameters, and off-diagonal elements denote the magnitude of cross-covariance between parameters. In the tie-line analysis, a nonlinear least-squares minimization is used, and the covariance matrix (Cov) is evaluated as Cov = r/H, where r is the residual and H is the Hessian at the solution. Errors in fitting parameters are propagated through the tie-line calculation, taking into account cross-covariance between fitting parameters whenever possible. In the three-phase analysis, linear least-squares minimization is used, and the covariance matrix is evaluated as Cov = r/(VV′), where r is the residual and V is the vector of best fit parameters. In the three-phase analysis, the fitting parameters are the desired endpoint compositions, and errors are determined directly from the diagonal elements of the covariance matrix. In practice, errors in tie-line and three-phase endpoints are dominated by the availability of reference spectra. Not surprisingly, the magnitude of errors depends strongly on the choice and availability of nearby reference spectra. When single-phase reference spectra are acquired near the desired tie-line endpoints, errors are significantly reduced (as demonstrated in SI Fig. 5).
Lineshape Fitting of Spectra Near a Critical Point.
MAS converts the intensity of 2H NMR spectra into a spinning centerband and spinning sidebands that appear on both sides from the centerband at multiples of the spinning frequency. Because the differences of chemical shift between resonances of carbon atoms C4-C15 are negligible, most resonances are superposed. At spinning frequencies in the range from 5 to 10 kHz, this signal is dominated by the resonances with the largest quadrupolar splittings, namely the chain methylene segments C2-C8 near the lipid–water interface (40). In Fig. 4 e, the measured width at half height of the MAS sideband is compared with the linewidth calculated from the measured R 2 rates of this same signal according to Δν1/2 = R 2/π.
The broadening of resonances near critical points is also visible in spectra obtained without MAS. The simulated spectrum shown in Fig. 4 f was generated by applying the line-broadening arising from R 2 rates measured without MAS to an order parameter profile obtained in membranes of the same composition but acquired at a higher temperature where quadrupolar splittings are well resolved (Fig. 7a). The R 2 rates were only determined for the largest quadrupolar splittings, corresponding to the so-called order parameter plateau in the spectrum (chain carbon atoms C2-C8). It was assumed in the fit that R 2 values at narrower splittings scale as the square of the order parameter, as expected from theory (30). In addition, all acquired 2H NMR spectra show evidence of a slight deviation from randomness of the orientation distribution functions of bilayer normals due to liposome deformation by the strong B o magnetic field. This was accounted for by fitting a bilayer distribution function to the experimental data (SI Fig. 7b ). In addition, the intensity of terminal methyl group resonances was attenuated to account for the experimental attenuation of those signals due to fast data acquisition (23).
We thank Ben Widom, James Sethna, John Cahn, Jenifer Thewalt, and Harden McConnell for helpful conversations, and Holger Scheidt for use of binary DPPC-d 62/Chol spectra. S.L.V. is supported through a postdoctoral fellowship from the Cancer Research Institute. S.L.K. acknowledges a National Science Foundation Career Award (MCB-0133484) and a Cottrell Scholar award from the Research Corporation. O.S. and K.G. are supported by the intramural program of the National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health.
- ¶To whom correspondence should be addressed. E-mail: gawrisch{at}helix.nih.gov
Author contributions: S.L.V., S.L.K., and K.G. designed research; S.L.V. and O.S. performed research; K.G. contributed new reagents/analytic tools; S.L.V. and O.S. analyzed data; and S.L.V., S.L.K., and K.G. wrote the paper.
↵ ‡Present address: Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/cgi/content/full/0703513104/DC1.
- Abbreviations:
- Chol,
- cholesterol;
- dioleoylphosphatidylcholine;
- DPPC-d62,
- chain perdeuterated dipalmitoylphosphatidylcholine;
- Lo,
- liquid ordered;
- Ld,
- liquid disordered;
- magic angle spinning.
- © 2007 by The National Academy of Sciences of the USA
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It is Veteran's Day in the US, and I thought we should look at the distribution of veterans across the country and North Dakota. To do this I grabbed some data from the Department of Veterans' Affairs. It includes both actual and projected veteran population data. First a look at the US as a whole.
Student blogging about the election
A student of mine, Richard Carpenter, has some interesting graphs and discussion of voting participation in the US compared to population on his blog. At various times in this election both candidates discussed the notion that they are brining new people to the voting booths, though I think we need to wait and see before declaring this some type of transformational election.
Population Changes in ND Counties
The 2015 ACS data came out from the Census Bureau in mid-September and gave us lots to chew on for the state of North Dakota. The four counties reported in the 1 year data are Burleigh, Cass, Grand Forks, and Ward. All three counties gained population from 2014. Cass increased the most with 4,507 more people, while Burleigh increased by 2,488, Ward by 1,891, and Grand Forks increased by 778.
OASDI and Workers per Retiree
I enjoyed my guest hosting duties on the radio today and wanted to follow-up on a question from a caller. The caller wanted to know how many people were currently working and paying into Social Security and if the number were lower than in the past and, as a result, causing some of the financial woes for the program. Actually the number of workers paying in right now is the highest it has been, but what I told the caller to pay attention to was the number or workers per retiree, or beneficiary. The OASDI trustees report (available here) gives all this information, but I boiled it down to a simple graph.
Fewer Shades of Grey in ND
JT tends to ask me the following question: "What worries me about the outlook for ND?" Over the last few years I answered that there was not much. That was then, this is now. North Dakota hit its first oil price catastrophe for this round of the oil boom and we are watching to see the consequences. One of the issues that worries me has to do with the demographics for the state as a whole. In this piece I will focus on the 65-and-over population and later on we can talk about other dimensions.


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The most common questions we get after our circus show are always, "What is your workout?" and "How do you stay in such good shape?" Well, we'd love to show you with this new creative project, a Circus Stella Workout DVD.
This dream has been a long time in the making. Being a gymnast as a child, and a professional circus acrobat as an adult formed the basis of my fitness foundation. But I needed to learn as much as I could about fitness and exercise science, so I chose to become a Certified Personal Trainer with the American College of Sports Medicine. After passing the exam, I found a mentor, fitness professional Mike Krug to consult on the DVD to make sure it will be the best quality workout and a useful tool for anyone who wants to improve their fitness. Add in qualified Yoga Instructor Amber Nedoff to consult on the stretching, plus the film crew from Video Dye Productions and you've got a great team to produce an outstanding workout DVD.
I want to help people find the joy in exercise and become fit, fitter, and circus fit! This DVD will include a full workout we can do together; warm up, training, cool down, and stretching. In addition I will provide my own tips on staying motivated and getting some extra movement into every day. And of course, we'll include a full performance of our show, Circus Stella, with rope-walking, aerial silk, trapeze, and performing rescued dogs! Yay dogs! Our dogs definitely help me stay in shape. Here they are helping me with my yoga...
Risks and challenges
I can be a bit of a perfectionist, so I'm sure what will take the longest is the planning and design of the exact workout plus creating lots of variations to meet everyone's different fitness level. The quality of the program is very important to me, so it will be a challenge to settle on the final design. Luckily I have great helpers in my corner who are willing to go over my plans and offer critiques and suggestions, like Mike Krug of Next Level Personal Training and Yoga Instructor Amber Nedoff.
The other potential challenge is the filming schedule. We'll have to film both during the Pennsylvania Renaissance Faire (to get the show footage) and on a day when they're closed (to get the workout). We'll be potentially limited by weather, as rain days would be impossible to film in, but luckily we have two solid months so surely we can find two sunny days to get everything on tape!Learn about accountability on Kickstarter
Funding period
- (29 days)


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Antoni Słonimski
Antoni Słonimski (15 November 1895 – 4 July 1976) was a Polish poet, artist, journalist, playwright and prose writer, president of the Union of Polish Writers in 1956–1959 during the Polish October, known for his devotion to social justice.
Antoni Słonimski
Antoni Słonimski
|Antoni Słonimski
15 November 1895
Warsaw, Congress Poland
|4 July 1976 (aged 80)
Warsaw, Poland
|Resting place
|Forest Cemetery in Laski
|Literary movement
Słonimski was the grandson of Hayyim Selig Slonimski, the founder of "ha-Tsefirah"- the first Hebrew weekly with an emphasis on the sciences. His father, an ophthalmologist, converted to Christianity when he married a Catholic woman. Słonimski was born in Warsaw and baptized and raised as a Christian.[1] Słonimski studied at the Academy of Fine Arts in Warsaw. In 1919 he co-founded the Skamander group of experimental poets with Julian Tuwim and Jarosław Iwaszkiewicz. In 1924 he travelled to Palestine and Brasil and in 1932 to the Soviet Union.
Słonimski spent the war years in exile in England and France, returning to Poland in 1951. He worked as contributor to popular periodicals: Nowa Kultura (1950–1962), Szpilki (1953–73) and Przegląd Kulturalny. He was an active anti-Stalinist and supporter of liberalization. Słonimski died on 4 July 1976 in a car accident in Warsaw.
- Sonety (1918)
- Parada (1920)
- Godzina poezji (1923)
- Torpeda czasu (Time Torpedo, 1926), a science fiction novel influenced by H.G. Wells[2]
- Droga na wschód (Road to the East; 1924), a collection of poems inspired by his travels to Palestine and Brazil
- Z dalekiej podróży (1926)
- Rodzina (Family; 1933), a comedy about two brothers: a communist, and a fascist
- Okno bez krat (1935)
- Dwa końce świata (Two Ends of the World; 1937), a novel predicting Warsaw's destruction by a Nazi dictator
- Alarm (1940)
- Wiek klęski (1945)
- Nowe wiersze (1959)
- Wiersze 1958–1963 (1963)
- 138 wierszy (1973)
See alsoEdit
- ^ Antony Polonsky, Monika Adamczyk-Garbowska (2001). Contemporary Jewish writing in Poland: an anthology. University of Nebraska Press. p. 321. ISBN 0-8032-3721-9. Retrieved 11 July 2011.
Source: "Alfabet wspomnień" by Antoni Słonimski (memoir), PIW, 1975
- ^ Patrick Parrinder, John S. Partington, The Reception of H.G. Wells in Europe. Continuum, 2005. ISBN 0826462537, (p. 140)
- Barry Keane, Skamander. The Poets and Their Poetry (2004), Agade; Warsaw, ISBN 83-87111-29-5.
External linksEdit
- Media related to Antoni Słonimski at Wikimedia Commons
- Profile of Antoni Słonimski at Culture.pl
|This article about a poet from Poland is a stub. You can help Wikipedia by expanding it.


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/ Medical Malpractice/ FAQs/ Does Having Insurance Affect My Settlement Amount in a Medical Malpractice Case?
Injury Attorneys | Restoring LivesTM
February 5, 2017 | Medical Malpractice | Social Share
Yes, your settlement amount may be affected if insurance covers all or part of your medical expenses. Insurance companies and healthcare providers can issue a lien against your settlement proceeds, which can lower the amount you receive in a settlement.
If an entity files a lien against your case, that entity has demanded reimbursement out of your settlement for the money it spent covering expenses after an injury.
A medical lien is a demand for repayment that can be placed against the compensatory outcome of your malpractice case. In other words, the hospital that paid for your medical expenses can demand those costs be reimbursed out of the amount you receive from your settlement.
Healthcare providers typically withhold from billing patients who are pursuing personal injury claims. They do so under the condition that the injured party will pay the claim with their settlement amount.
Liens can take several forms:
You have an obligation to satisfy the lien by paying it out of your settlement. Insurance companies often seek reimbursement for medical costs directly from the settlement, a process called subrogated interest.
The amount taken out of the settlement is dependent upon the nature of the costs fronted by an insurance company or healthcare provider and the nature of the lien. Many liens are negotiable, and even disputable. Hospitals often agree to accept less than the full lean amount, waiving the balance owed.
In the state of Indiana, liens cannot be filed against someone who is receiving Medicare or Medicaid benefits.
If you or a loved one have been a victim of medical malpractice and are unsure of how your health insurance will affect your lawsuit, you are urged to contact the Indianapolis Medical Malpractice Attorneys of Wilson Kehoe Winingham. The lawyers at WKW can help you get the compensation you deserve. Call 317.920.6400 or fill out an online contact form for a free, no-obligation case evaluation.


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Future Bouffe
(Photo Bruno de Nazareth)
The élite have at their disposal a range of resources that are broader, healthier, and more sophisticated than does the population as a whole. At the pinnacle of this hierarchy, certain practices now in large-scale development breach even the meagerest remaining levee of what once was called professional ethics. Please find this issue explored in detail under the heading 'Privatization of food resources and human capital' (p. 11)…
— Jean-Roger Helmin, MANGER EN / NUTRITION IN 2050, Éditions C. F. A.
I translated the fictional journalist's work for this bilingual edition, exhibited in Paris and Lille.


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by Benson Agoha
Russia yesterday said it was "extremely concerned about the development of the situation in Ukraine."
In an official statement Russia's Foreign Office said the armed confrontations between groups and law enforcement agencies was of serious concern because agreement reached on 21 February was ignored, even though it had been midwifed and welcomed by internationally eminent foreign representatives.
"There have been armed confrontations between violent youths, extreme right nationalist organisations and units of law enforcement agencies, who defended peaceful civilians and interests of the state, in the capital and several other cities recently."
"The agreement on settlement of the crisis in Ukraine of the 21 February is not observed despite the fact that its signature was certified by Foreign Ministers of Germany, Poland and France, as well as the United States, the European Union and other international bodies welcomed this document."
Russia said "Militants have not been unarmed, they refuse to leave the streets of cities, which are actually under their control, refuse to free administrative buildings, continue acts of violence."
The statement further said, "[We are surprised that several European politicians have already sprung to support the announcement of presidential elections in Ukraine this May, although the agreement of the 21 February envisages that these elections should take place only after the completion of the constitutional reform. It is clear that for this reform to succeed all the Ukrainian political forces and all regions of the country must become its part, but its results should be approved by a nationwide referendum. We are convinced that it is necessary to fully take into account concerns of deputies of eastern and southern regions of Ukraine, the Crimea and Sevastopol, which were expressed at the conference in Kharkov on the 22 February.
We are deeply concerned about the actions in the Ukrainian Verkhovna Rada in terms of their legitimacy. Actually referring to the "revolutionary appropriateness" only, they are stamping "decisions" and "laws", including those aimed at deprivation of humanitarian rights of Russians and other national minorities living in Ukraine.
There are calls to prohibition [sic] the Russian language almost fully, lustration, liquidation of parties and organisations, closing of undesirable mass media, removal of restrictions for propaganda of Nao-Nazi ideology.
The course is to suppress those, who do not agree to this, in different Ukrainian regions by dictatorship and even terrorist methods.
There are threats to Orthodox sanctities.
National radicals continue to scoff at monuments in different Ukrainian cities, while like-minded persons in some European countries besmear memorials to Soviet warriors.
Such development of events disrupts the Agreement of the 21 February, discredits its initiators and guarantors, and creates a threat to civil peace, stability in the community and safety of nationals.
We are forced to note that some of our western partners are not concerned about the fate of Ukraine, but rather their own unilateral geopolitical considerations. There are no principled assessments of criminal actions of extremists, including their Neo-Nazi and anti-Semitic manifestations. All the more so, such actions are intentionally or unintentionally promoted. We cannot but get a sustainable impression that the Agreement of the 21 February with silent consent of all its external sponsors is used as a cover only to promote the scenario of change of Ukrainian power by force through the creation of "facts on the ground", without any wish to search for a Ukraine-wide consensus in the interests of national peace. We are especially worried about the attempts to involve international structures, including the UN Secretariat, into the approval of this position.
We insistently appeal to all those who are part of this crisis in Ukraine to demonstrate maximum responsibility and to prevent further degradation of the situation, to return it to the ambit of the law, and to decisively stop those extremists, who are seeking power,]" the statement said.
The Agreement of 21 February, 2014, was welcomed by Prime Minister David Cameron and several other well meaning governments, before the turn out of events that reportedly led to the death of some 70 to 100 people, the President Viktor F. Yanukovych rejected it, triggering further violence. He has now been impeached.
Ukraine's violence started late in 2013 after President Yanukovych moved to align Ukraine more closely with Russia, while the opposition wan the country to join the European Union.
* Twitter: @bensonagoha.
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- Property Taxes: $1,777
- Property Tax Year: 2017
- Listing Company: Rieckmann Real Estate Group, Inc


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Conor McGregor has revealed he's ready to put on the boxing gloves once again to take on Floyd Mayweather, and this time he says 'I know I would win'.
If we cast our minds back to 2017, we'll remember McGregor said the same thing in the build up to his initial fight with Mayweather. But, unfortunately for the retired UFC fighter, it all got too much in the 10th round when the referee was forced to stop the match.
Now, two years on, McGregor says he's confident he's got what it takes to give Mayweather his first professional loss.
Speaking on Saturday's episode of the Tony Robbins Podcast, he said:
I would like to rematch him under boxing rules, again. And I believe I would win.
Actually, there I go again with that fake humbleness. I know I would win.
Looking back, McGregor doesn't see his 2017 fight as being Mayweather's usual cat-and-mouse game he's so famous for, and actually believes he was winning the match in earlier rounds.
He said:
When I went into the fight in the early rounds I was whooping him in the early rounds. I went back to my corner after the first round and I said, 'This is easy!'
Instead, he believes Mayweather switched his technique mid-way through the fight, to a style he's never used before. McGregor also believes it was his aggression in the first half that forced Floyd to change his style.
He said:
He had to switch up his style, and you've got to respect that, he switched to the Mexican boxing style.
The Mexican boxing style is where you put your elbows in tight and walk forward with heavy pressure. He never fought like that in his entire career.
He never fought like that in his entire career, but he was forced to fight that way because he was getting beaten.
The approach caught me off guard. I was not prepared for it, I was not used to it, and he walked me down and ended up getting the stoppage.
Now, McGregor has re-joined the list of fighters hoping to coax semi-retired Mayweather back into the ring, alongside the likes of Manny Pacquiao, who frequently talks of his desire to avenge his final loss against Mayweather.
Could he be tempted? Well, considering he's reported to have made $275 million out of his fight against McGregor, we'd never say never.
If you have a story you want to tell send it to UNILAD via [email protected]
Emma Rosemurgey is an NCTJ trained Journalist at UNILAD. She graduated from the University of Central Lancashire in Preston and started her career in regional newspapers before joining the LADbible Group team in 2017.


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Low Top
Extremely popular sneakers because of their price range, versatility when it comes to style, and freedom of movement. Almost everyone is guaranteed to have low-top sneakers in their shoe rotation.
Mid Top
Mid-top sneakers extend toward the ankle for a little more support and hold. These lie somewhere in the middle between low-top and high-top sneakers in terms of usage and popularity.
High Top
Sneakers with collars that go above the ankles for optimal hold and support are some of the most sought-after models in lifestyle shoes. Most of these shoes take their roots from basketball and have easily or fashionably crossed to mainstream wear.
Good to know
Regardless of cut, it's always good to start with sneakers that can be worn for the daily grind, also called as "beaters" by some, as these are usually cheaper, easy to clean, and still gives that lifestyle "edge" before going for those wallet-thinning models.
Inspired from
Sneakers dominantly take their heritage from running, basketball, skate, tennis, training, hiking, and football. Still retaining a few of their performance-based technologies, these sneakers have transcended their respective niches and have successfully and popularly transformed themselves as staples of fashion footwear.
Sneakers designed for a laidback, "cool" vibe that is built for lifestyle wear right from the get-go.
Good to know
Brands are now blending elements of performance and casual appeal in basically every sneaker. One can hardly go wrong with a sports-inspired sneaker or a simple casual shoe.
Good to know
Shoes sharing the same inspiration, history, materials, or technologies are routinely assembled under one compilation for the convenience of those who may wish to categorize or label their own collection as such. The classic collections like the Adidas Originals, Air Max 1, Air Force 1, new balance classic sneakers, and the Classic Leather head the pack of frequently asked about collections.
The VS Hoopster Mid is available in women's sizes. Sizes offered range from 3 to 11. Since the shoe is made up of synthetic leather, it's important to note to get your true size since it won't stretch as much as real leather does.
The understated sporty casual style of the Adidas VS Hoopster Mid is perfect for weekend trips around town or hanging out with your friends. Wear this mid top footwear with leggings, jeans, or distressed denim topped off with a cropped top or varsity shirt, and you're all set.
The Adidas VS Hoopster Mid's clean, basketball-inspired silhouette is easily its most distinguishable feature. The grooved texture on the shoe's quarter, enamel finishing on the three stripes, and patent finish on the toe cap gives this footwear a fresh and young look.
The VS Hoopster Mid is a women's shoe that is part of the Adidas NEO collection. Neo stands for new definition, concept, and spirit. The line was launched first in Germany in 2012, with the opening of ten NEO retail stores. The target market of the line is young consumers, primarily teenagers who have average spending powers.
NEO is Adidas' fast fashion retailer sub-brand that is known for delivering new inventory to their stores six times per week, ensuring an endless supply of new products for their young market. The Neo label is an apparel-driven range but also includes footwear and accessories.
The VS Hoopster Mid is one of the NEO footwear models. The mid top has a subtle style that is rounded off with an enamel-finished three stripes and a synthetic upper with embossed and perf details.
- The shoe features a comfortable textile lining and sockliner, a rubber cupsole, and synthetic leather upper.
- Weight is around 315 g per shoe.


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Mitt-17 cm x 30 cm / Pot Holder-20 cm x 20 cm
Take your baking experience two steps further. Pull out freshly bake cake with the Ellementry Mitt and Pot set.
Wash it at 30° in the washing machine. It's good to iron.
Have you bought the blue and multicolour classic mitt and pot holder?
Let us know what you think.


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Britons splash cash on going out rather than buying new clothes
Spending on holidays and going out boomed last year as British consumers chose to splash their cash on entertainment over refreshing their wardrobes.
Consumer spending grew by 3.6pc between January and November compared with a year ago, according to Barclaycard.
While this is slightly down on last year's growth of 3.9pc, Barclaycard, which processes nearly half of all UK credit and debit transactions, said spending on the "experience economy", including trips to the cinema, restaurants and theatre, saw robust growth throughout the year.
It noted that households returned to "business-as-usual" following the UK's decision to leave the EU, with the referendum result having "no short-term impact...
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AS411 Community Home » Accounting Software » General Discussions » Collections Newbee
I am currently working as a receptionist. My boss has asked me to make collections calls. (She thinks it falls under administrative duties)I am not sure what it may entail.I made calls at my last job as an office manager/accountant but i was never trained in collections. That job was for a small manufacturing company. I now work for a multimillion dollar software company and I am sure the approach and client facing would be much different.any hints or tips would be much appreciated....
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Recent posts
Chapter Two: A White Century
(H20, Kazuya, and Florina) The chainsaw releases a sucking noise as it is torn out from the captor's chest leaving behind a gaping hole that continues to bleed. The captor's corpse falls on the lap of the inattentive girl, decorating her white robes in red. Yet the girl does not react to the sudden weight of the captor, but rather the attempt made to seize the device. As soon as the device is moved from its spot a pale hand is latched to the body of it and the hand is strong enough to prevent further movement of the device. The girl's eyes hone in on the person snatching the gun. Her person is still devoid of any reaction, but her blank gaze carries a threatening aura. Byakuran-sama will not be pleased if you take one of his toys. Is all she says as orange flames start seeping out of her body in a predatory manner. Meanwhile, the demonic creature swims to safety in its watery prison as terrible gusts of wind slice through its abdomen. Water burns at the creature's body ripping off black flakes from its skin. No blood is to be found as the creature is dissolved from head to toe. What is left of its body doesn't make it close to the edge of the prison before giving away completely and releasing dark indigo smoke. However, the chainsaws used by the creature remain on the floor and the demon mask it had sits at the base of the liquid prison. (Toki and Liam) After some careful movement, you find yourself at the base of a statue carrying two ivory dragon locked into ferocious combat. One of the dragons is as white as fresh snow and massive angelic wings stretch out from its rigid back as if to wrap the other dragon in feathers. While the other dragon is as black as night and fangs that seem to bleed in the light as they sink into the white dragon's throat. The two dragons appear to be of the same species with similar limbs and lithe forms, but their frozen combat is brutal and cruel. They both lack chunks of their body that could only be clawed out by fang or talon and their mouths are curved back in expressions of sheer rage. A plaque at eye level on the wall behind the statues reads. The pride of the Millifiore Famiglia Our leader in two forms His constant conflict brings humanity to new heights Exitus Acta Probat You and your companion are left in peace to read the scripture on the plaque, but not a moment too soon all the traditional lights go out and the room is bathed in darkness. Only the stars beyond a ruined portion of the ceiling can be seen as the air suddenly chills.
Chapter Two: A White Century
(Bartz) Beneath the portrait you find a golden tag with fanciful words written in cursive. Uni Boss of the Giglo Nero Famiglia Third Sky Arcabaleno Loving daughter, leader, and boss Are what the words read. The tag covered in a fine layer of dust with a small cobweb attached to it. Upon closer inspection you notice that the girl in the portrait has a face still shedding off the dredges of youth, but her expression is one that stares off into the distance. Her eyes are wide, innocent things while her clothing lies on the mature side in the form of a black dress. Even the illusion of her tiny stature is ruined with the regal framing of the portrait. With every sign of youth seen upon the girl there is another detail in the art that screams power and responsibility. (Toki) You follow after the companion that has taken to peeking into places he doesn't belong and find yourself in an abandoned wing of the manor. Up ahead you can see your companion intensely studying a portrait depicting a young girl in mafia garb. While to your left is the servants door and to the right are the dragon statue hiding an additional door behind it. You and your companion are the only two occupying the wing, but you can hear servants stir behind one of the doors. (Quintus) Sorry, I am a bit of a mind reader so sometimes information just comes to me~ Stuff like your name, the names of those that accompany you, your darkest secrets, what you fear, first love ... all of that floods in sometimes. I hope you don't mind this little old curse of mine~ Byakuran playfully tells you this as he picks off a stick covered with marshmallows off the tray of a server. Some snacks my dear? He holds the stick out in mock offering before taking a bite of it himself. Satisfied with the treat he procured for himself, Byakuran focuses again and is blunt with his follow up. I can give you some business right now Quintus. How about you help me with the executions I have planned for tonight? Byakuran moves to the edge of the balcony and rests himself against the marble barricade that separates him from the rest. A quick look to your direction is the most he gives you to indicate that he wants you to follow. Horrible human beings the lot of them.
Chapter Two: A White Century
(Kazuya, Florina, and H2O) The captor obediently bends to pixie's command and simply drops the device on the table. His lack of care when dropping the device manages to shatter the glass container attached to it. The black liquid within slips through the remains of the container, greedily eating into portion of the table. Ignorant of this the captor jabbers on revealing plenty of tidbits that could be useful. Byakuran is the boss of the Millefiore famiglia and a wielder of the almost extinct sky flames. With his flame type he can easily rule over anyone residing under the sky dominion, but that isn't what makes him so great. Conspiratorially, the captor leans toward pixie and adds. Byakuran can see multiple parallel worlds at once. He can fix our world by seeing what went wrong in other versions of it. The captor would have said more if he did not have a chainsaw ripping straight through his sternum. While the captor was talking, the demonic faced creature had been tossed into the air and promptly manhandled by tendrils of water. However, not heeding the given warning the creature had struggled against the tendrils until the water sank into its flesh and bone. The tendrils burned through the creature's robe revealing dried, cracked skin covered in talismans. Yet the creature offered little reaction to the punishment as it allowed portions of its arms and legs to be cut off by the tendrils. Not even a whimper of pain escaped the creature's mouth as its limbs were effectively severed into tiny pieces. Instead the portions of the limbs that were cut off easily turned into purple smoke as the creature regrew its missing limbs before your very eyes. Purple smoke hardening into what was lost and the creature attaching the perfect facsimiles to its body with a sickening snap. Once the creature finished fixing itself it had then struck at its next target: the captor. Appearing from behind and shoving one of its powerful chainsaws into the captor's chest with brutal efficiency. You will now die. Is all you hear over the deafening noise of the chainsaw.
Chapter Two: A White Century
(Florina, Kazuya, and H20) The captor is easily charmed by Pixie's wiles and with little hesitation says. I was ordered by Byakuran to sedate the sky arcabaleno for the main event. Upon stating this the captor does look at the tool in his hand with clear intent to continue his job. However, his lax focus prevents him from really continuing with the task and the tool simply hangs on the hand in a loosening grip. Byakuran won't be made at me for entertaining such a charming lady, right? The captor asks Pixie with a relaxed grin. Up until now the demon masked creature had been quietly sitting across the girl, but upon hearing the question of the captor it suddenly dissipates in form. A few seconds later it reappears behind you and the roar of chainsaw pounds in your ears with the proximity of its source. A chainsaw is aimed at the abdomen of the one with guns.
Chapter Two: A White Century
(Quintus) The man pulls back his hand and momentarily frowns. You might not be one for entertaining, but an ally of yours seems to know how to grab my attention. He then pulls back the left sleeve of his coat to look at a watch on his previously covered wrist. Humming, he looks to the tiny hands within before letting the sleeve reclaim his wrist. You should introduce me to Toki when you get the chance. A gentle laugh escapes the man's lips. Shame on you for keeping me to yourself. I can be shared. The man reconsiders his statement right after it is said. Well as long as you continue to have anything interesting to bring. Byakuran tis I, but tell me who are you "Quintus"? Byakuran puts some stress on your name with a conspiratorial glance in your direction. (Bartz) Peeking inside you see a darkened wing of the building that hasn't had a good sweep in a long time. Dim flames flicker out from ancient candles mounted along the walls as slight bursts of cold wind quick up dust in steady intervals. Right next to your place of peeking you can see a door marked as the kitchen. Light emanates from the space between the door and the floor giving some life to the rest of the wing. The other two doors connected to the wing are dark and cold in an ornate manner. A portrait of a young girl with bluish hair hangs alongside one of the doors at the very end of a line of portraits depicting women of similar characteristics. While the other darkened door is almost hidden behind a deteriorated statue of two dragons battling it out in suspended air. Servants have stopped flying through the kitchen doors for now, but there is no telling when they will be called on again. (Toki) The messenger nods in response before turning back to clean his pool stick. You arrive back to the grand ballroom in the midst of a lagging portion of the party. Folks that were previously dancing are now taking seats alongside the party floor to relieve their weary feet. While those that partook in general consumption wander about aimlessly without any acute awareness to their surroundings. Some even rest against the walls for support while recovering from the highs they submitted themselves too. All this makes for a cleared hall and you can easily spot one of your companions engaging with the white haired man seen earlier. While another companion can be seen openly peering through a door at the side of the hallway. Guests whisper and look at the odd behavior of this companion, but none seem willing to engage with him yet. You are unable to spot anyone else from your group from where you are.
Chapter Two: A White Century
(H20, Florina, and Kazuya) The captor leaps with surprise upon being addressed. As the needle delicately placed on the girl's neck is shifted with his sudden hand movement. The needle is sharp enough that the momentary slip is enough to draw blood from the girl's cream neck. Before the captor can do anything to amend his error some blood spills onto the girl's lap and blends with her black uniform. Despite the mistake made the girl does not react at all to your sudden intrusion or the cut made on her neck. At best she absently scratches the cut drawing more blood and doesn't stop until her captor forcibly hold hers hand. Her gaze remains focused on the wall across. As the demon faced entity below it also remains stoic and unreactive. The only one really attentive to you is the captor and his response is short and sharp. Just administering some medicine. Now if you could please leave so I can continue. The captor curses under his breath after answering and quickly opens the pocket of the white cloak. From it he obtains a capsule containing a dark liquid blend with flames racing upon its surface. He replaces a capsule of similar substance with the new one and then refixes it to the machine.
Chapter Two: A White Century
(Bartz) Tracing back your steps leads you past the side hall connecting to the garden and up the balcony where you originally started. Though the increased attention you pay does allow you to notice a smaller additional door along the right hallway of the room. The small door continually flings in and out as mobs of servants rush through it to maintain the opulence of the gathering. A silverette with a tux and a few too many rings bumps into you as you watch. Sorry. He barks out. Before taking up conversation with a fair lady in a rosy dress. A skull-shaped ring is left behind after the sudden collision. (Toki) The man studies you with an odd expression. You really came to the greatest mafia gathering ever held without any idea of who was running it? Within seconds you find a pool stick aimed directly at your chest. The man watching in a hungry manner as his left hands firmly grips the stick and aims it at you. Byakuran is the main star here. But anyone that belongs here would know that. Air shifts around the tip of the pool stick as the man sizes you up with his eyes. Show me that invite of yours missy. (Quintus) You reach the secondary balcony in the ballroom and what awaits you there is the same man that beckoned you earlier. Up close you can tell that his white hair is a veritable bird's nest and his pristine white uniform had been tossed on without a care in the world. Easily separating this man from the elegant people around that hunger for his attention. Yet, in mannerisms this man holds himself in a posture that naturally begs authority. He almost towers over you on the stairs without even intending to. Though there is something missing from his stance and the air around him feels empty without it. The space just simmering with the lack of something. This master does not care much for the empty space surrounding him. Instead greeting you with the bubbly smile he offered earlier and sharp eyes that seize every detail on your person. Have you come to entertain me outsider? The rest on the balcony watch you through masks that betray no features.
Chapter Two: A White Century
(Gamma) The seal splits in two revealing a slip of paper within the envelope that is scrawled upon with messy, rushed writing. When read the letter urgently states. This life resets at midnight. You only have a few hours to resolve things. Before all the pieces are moved back to their original positions. The reset will set you back. Please hurry! - Favoring The man allows you to read the letter without interruption. As you reach the last line he finally breaks the silence by tossing a golden pocket watch at you. The desperate young lady also wanted you to have this. Said that it was very important. Probably a family heirloom or something. If you were to open the watch you would see two golden needles indicating that it is nine. (Quintus and Liam) You take the left path in the grand ballroom and travel to meet the man with white hair. Along the way there is an arched exit to the side that leads to a garden. However, the darkness outside makes it difficult to pick out any distinguishing traits of the two people currently standing out there. No one else is entering the garden at this time making it a sort of dead zone among such a vibrant display of human antics. Meanwhile, the white haired man is staring at you and awaiting your arrival on the furthest balcony. His location is up ahead while the garden would veer you off a side path. (Florina, Kazuya, and H2O) You pass the rustic door and enter into a massive room that could fit thousands of people within. It should be impossible for a room of this stature to exist beyond such an unassuming wall, but here it is in full view. Adding to the room's oddity is its pure white color scheme. Everything is bathed in white from its walls to its flooring and any decorations in between. Even the vases and portraits are colored as if some child went liberal with the white paint. The only thing lacking in magnanimity and white is a traditional dining hall table that stretches from a few feet near you to across the huge room. The brown sheen of the table makes it stand out from everything else around you and the chair against it act as little brown dots in the white desert. Seated nearby on one of the chairs is the blue haired girl you decided to follow and her captor stands behind her. In her captor's hands is some device that you can't make out, but you can see that he is rubbing gauze on her neck in preparation. The girl doesn't seem to mind as she continues chewing on her stolen snack and staring blankly at the white walls across from her. The white cloak that she had been wearing is laid on the table. Across from the girl and her captor sits an entity with a red oni mask on its face and hooded grey cloak. The entity pokes at the fabric on the table seeming as disconnected from the world as the girl.
Chapter Two: A White Century
(H20) You catch up with your two companions just in time to watch one of them leave to follow a girl. The incident you helped cause in the antechamber seems to be going ignored for now as the music drones out any concerning noises. But, that may not last forever though the drunken state of most of the people could be to your favor. Many are so befuddled that they take your suit for costume and poke fun at you to their plus ones. One tiny girl in black uniform even tries to grab at you as you pass and stares at you with huge, awestruck eyes. Whispering. Hero-chan. As you move along. (Florina, Kazuya, H20) With one leading and the other two carefully following the hunt for the captured maiden begins. Though the task isn't easy as her captor quickly drags her through the crowds with a skill that only someone used to parties can have. The maiden doesn't provide much resistance to the dragging either as she leans into the hold allowing her white cloak to trail along the spotless floor. Her dull eyes look to nothing in particular as she munches on a donut that she managed to grab before capture. Eventually, the captor and the maiden exit the side hall and enter an old wooden door instead of approaching the crowded balcony. The white haired main pauses in his festivity as he watches the two go and then he stares at you with pure curiosity. Waiting to see what happens next in this slipshod night. A rustic, wooden door heading into the depths of the manor or an open balcony visible to all yet holding the shadiest of characters. The white haired man doesn't say anything to rush a choice, but the crescent moon ascending behind him says otherwise.
Chapter Two: A White Century
(Quintus) A few look to you with pity and others pointedly glare at you with sheer hostility as you meekly wander about. No one lifts a hand to help as all attention is captured by the festivities of the night. Your way forward is blocked off by one masked stranger after another until you are practically pushed against the marble railing of the balcony. Beneath dancers continue to sway and weave in haphazard harmony. While flames of various colors dance from the barest of skin or shred of cloth upon the person. As the dancers light up to a climbing crescendo your eyes are met with pale, purple orbs across the expanse of the room. Etched into the pale face of a content man these orbs wink at you as if in understanding. Beneath the left one is a purple tattoo that is jarring to the sight as a few strands of white hair fall over the other. After winking, the man waves to you with a bubbly smile. (Bartz) Tracing back your steps to the entrance of the room you soon find a companion of yours being pushed against the marble railing of the balcony. He seems to be slightly anxious, but the party goers pay him little attention as they get rowdier and rowdier. The force held in the newest song being played does little to help as people unwittingly match the energy of the music. Nearing your companion you notice that the white haired man across the balcony seems to lose interest in the dancers. Turning his back completely to you and your companion once you arrive. (Toki) You enter a garden bathed in the light of a crescent moon and filled to the brim with white orchids of various temperaments. The terribly white flowers look almost ethereal under the night sky as calm winds gently sway them apart petal by petal. It is here in this shedding of white that you find a man in black uniform holding a plain letter in his hands. The man wears a plain white shirt with a black coat over it as the rest of him remains demure in appearance. The most unusual thing he carries on him other than the letter is a pool stick, but even that is lacking in any unique inspiration. Not that the man seems to mind as he allows the wind to mess with his hair and a few petals land to land on the yellowy strands. He only motions to fix himself once he notices you arrive unto the grass. I believe someone left this for you. The man holds out the letter in his possession. The white envelope betrays nothing of value, but the red seal keeping it closed resembles a a wolf in hunt. It's dangerous getting admirers in places like these. Before you know it the girl has a dad with thick wallets and plenty of men willing to shoot a bullet.
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Chapter Two: A White Century
You are abruptly dropped with your companions at the center of a large gathering of people. Gone is the pretty spacious elevator from which you entered, now replaced with a high profile antechamber decorated in the finest of luxuries. From where you have fallen you can see ostentatious couches, marble animal statues, and paintings dating back to a classical era. The calm melody of a fine German piece of music waft in accompanied with a wiff of orchid and rose. The soft peals of water from a fountain within the room sometimes overpowers the music, but both sounds never become too overpowering. Laughter and light bits of conversation are also sprinkled about from the mouths of people wearing vibrant, decorative masks. Upon your arrival some of the people had filtered around you and your companions in a circle and are now discussing this random occurrence in harsh whispers. Their masks occasionally leaping up with their facial movements and their fancy, frivolous style of dress putting what you are wearing to shame. These people don't even bother to help you get up as they chat on and on around you. At the furthest end of the antechamber is a regular sized door decorated with perfect white orchids. It would have been easy to reach on a different day, but the circle of people and the rest walking about could easily slow you down.
Intermission One
(Toki, Alphinaud, Cait, Pancake Knight, Niko) You fall out of the elevator you originally entered and land on a heap of your own comrades. A pomegranate rolls away from the pile and lies within arms reach. If you were to look around you would see the faces of the people you previously left behind. In front of you stands Walton with an unreadable expression on his face. Around you is the lobby from which you started your journey. The only object left from the trip is the lone pomegranate on the floor. (Everyone Else) You are able to see your formerly gone companions drop out from the elevator. They seem intact and what they started with is there, but one companion is missing. The screens that let you observe the journey shut off for now.
Chapter One: The Philosopher King
(Pancake Knight, Cait, Sam, and Toki have arrived) You materialize along with three others in the middle of a crowded marketplace. There are no traces of the elevator left and the surrounding mobs remain blissfully unaware of your sudden appearance. The general pushing of the crowd indicates that you are tangible, but no one raises a fuss. If you were to look around you would see plenty of happy people with impossible shades of hair going about their day. Each person strolling from one wooden stall to the next and checking out the available wares. There is also an apparent trend among the people of wearing simple or elaborate robes paired with sandals. Some of the robes are equipped with various symbols or items of jewelry making you and the others stand out like sore thumbs. Overhead the sun casts an oppressive heat on the place making the sea of white tarps above the stalls almost glow. While the brown adobe buildings behind the stalls look paled in the morning light. The sun has the most effect on a tall, glass tower in the distance which is almost painful to gaze directly at. Not that anyone does so already preoccupied in the mundane rituals of the day. At the end of the marketplace are a flight of stairs heading up from between the buildings. However, the stairs are occupied by a swath of men decorated in scaled armor with rapiers at their sides and a tall humanoid dragon-like man.
You are stumbling ... Dropping ... Falling from high above. With no clear recollection of where you had been or where you will be going. You feel lightheaded as if freed from all restraint. Though what feelings this sensation could cause is best left up to the one experiencing it. Meanwhile, around you are cascades of words, letters, and random phrases tumbling to the unknown. You follow the same path but at a slower less rushed pace. Plummeting calmly enough to make out meaningful words or details that should jog your mind a little. Pray tell what could you be seeing in the maelstrom of words? (All your actions and choices will be etched in stone)
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Tesla Announces Milestone Achievement: Model 3 for $35,000
Model 3 is a smaller, simpler and a more affordable electric car. Designed and built as the world's first mass-market electric vehicle, it is a critical step in Tesla's mission to accelerate the world's transition to sustainable energy.
Like every Tesla vehicle, Model 3 combines range, performance, safety and technology. Intelligent design maximizes interior space to comfortably fit five adults and all of their gear. The high-efficiency powertrain provides zero to 60 mph acceleration in as little as 3.2 seconds.
Model 3 is designed to achieve the highest safety ratings in every category. With 220 to 325 miles of range and a starting price of $35,000 before gas savings and incentives, Model 3 is Tesla's most affordable vehicle yet.
Model 3 deliveries began on July 28, 2017 at the Tesla Factory in Fremont, CA. It is currently available for purchase on Tesla.com in a number of different configurations, including Standard Range, Mid Range, and Long Range battery options. Certain variants are available with Dual-Motor All-Wheel Drive for superior performance and traction in any weather, and Model 3 Performance is also available for best-in-class acceleration. Please visit our Design Studio to see available options.
Dimensions & Weight
- Length: 184.8"
- Width: 82.2" (76.1" with mirrors folded)
- Height: 56.8"
- Wheelbase: 113.2"
- Track (wheel center): 62.2" front and rear
- Ground clearance: 5.5"
- Head room, standard: 39.6" front row, 37.7" second row
- Head room, glass roof: 40.3" front row, 37.7" second row
- Leg room: 42.7" front row, 35.2" second row
- Shoulder room: 56.3" front row, 54.0" second row
- Hip room: 53.4" front row, 52.4" second row
- Seating capacity: 5 adults
- Luggage capacity: 15 cubic feet
- Weight distribution:
- 47% front, 53% rear (Model 3)
- 48% front, 52% rear (Model 3 Mid Range and Long Range RWD)
- 50% front, 50% rear (Model 3 Dual Motor and Performance)
- Hybrid steel/aluminum body
- Drag coefficient of 0.23
- Double wishbone, virtual steer axis front suspension with coil over twin-tube shock absorbers and stabilizer bar
- Independent multi-link rear suspension with twin-tube shock absorbers and stabilizer bar
- Variable ratio, speed sensitive electronic power steering
- Electromechanically boosted four wheel anti-lock disc brakes with electronic brake force distribution
- 18" aero or 19" sport or 20" performance wheels with various tire options
Standard Accessories
- 20 foot mobile connector with storage bag
- 240 volt NEMA 14-50 adapter
- 120 volt NEMA 5-15 adapter
- J1772 public charging adapter
Charging Rates
Standard Battery
- Supercharging rate: 150 miles of range per 30 minutes
- Home charging rate: 37 miles of range per hour (240V outlet, 40A)
Mid Range Battery
- Supercharging rate: 150 miles of range per 30 minutes
- Home charging rate: 37 miles of range per hour (240V outlet, 40A)
Long Range Battery
- Supercharging rate: 170 miles of range per 30 minutes
- Home charging rate: Home charging rate: 37 miles of range per hour (240V outlet, 40A)


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Evaluating Investments
For the financially sophisticated, we are able to provide sound investment advice, no matter what your stage of life.
Whether you want to buy into an existing business or expand what you already have, we are able to help you look at the costs and benefits of doing so.
We can evaluate capital expenditure or staff employment decisions and review proposed geographical or cross-industry expansion plans, including vertical integration with either your suppliers or customers.


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Country Montessori Preschool of Sutton
Education / Elementary and Secondary Schools
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Country Montessori offers young children a distinct learning experience using the philosophy and methodology developed by Dr. Maria Montessori. Dr. Montessori believed that children are natural learners and that between the ages of birth and six, the mind develops faster than any other period in life.