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Chemical Sciences: A Manual for CSIR-UGC National Eligibility Test for Lectureship and JRF/Fluorescence correlation spectroscopy
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Fluorescence correlation spectroscopy (FCS) is a common technique used by physicists, chemists, and biologists to experimentally characterize the dynamics of fluorescent species (e.g. single fluorescent dye molecules in nanostructured materials, autofluorescent proteins in living cells, etc.). Although the name indicates a specific link to fluorescence, the method is used today also for exploring other forms of luminescence (like reflections, luminescence from gold-beads or quantum dots or phosphorescent species). The word "spectroscopy" in the name differs from common usage, in which a spectrum is generally understood to be a frequency spectrum. The autocorrelation is a genuine form of spectrum, however: It is the time-spectrum generated from the power spectrum (via inverse Fourier transform).
Commonly, FCS is employed in the context of optical microscopy, in particular confocal or two-photon microscopy. In these techniques light is focused on a sample and the measured fluorescence intensity fluctuations (due to diffusion, physical or chemical reactions, aggregation, etc.) are analyzed using the temporal autocorrelation. Because the measured property is essentially related to the magnitude and/or the amount of fluctuations, there is an optimum measurement regime at the level when individual species enter or exit the observation volume (or turn on and off in the volume). When too many entities are measured at the same time the overall fluctuations are small in comparison to the total signal and may not be resolvable – in the other direction, if the individual fluctuation-events are too sparse in time, one measurement may take prohibitively too long. FCS is in a way the fluorescent counterpart to dynamic light scattering, which uses coherent light scattering, instead of (incoherent) fluorescence.
When an appropriate model is known, FCS can be used to obtain quantitative information such as
- diffusion coefficients
- hydrodynamic radii
- average concentrations
- kinetic chemical reaction rates
- singlet-triplet dynamics
Because fluorescent markers come in a variety of colors and can be specifically bound to a particular molecule (e.g. proteins, polymers, metal-complexes, etc.), it is possible to study the behavior of individual molecules (in rapid succession in composite solutions). With the development of sensitive detectors such as avalanche photodiodes the detection of the fluorescence signal coming from individual molecules in highly dilute samples has become practical. With this emerged the possibility to conduct FCS experiments in a wide variety of specimens, ranging from materials science to biology. The advent of engineered cells with genetically tagged proteins (like green fluorescent protein) has made FCS a common tool for studying molecular dynamics in living cells.
Signal-correlation techniques were first experimentally applied to fluorescence in 1972 by Magde, Elson, and Webb[1], who are therefore commonly credited as the "inventors" of FCS. The technique was further developed in a group of papers by these and other authors soon after, establishing the theoretical foundations and types of applications.[2][3][4] See Thompson (1991)[5] for a review of that period.
Beginning in 1993[6], a number of improvements in the measurement techniques—notably using confocal microscopy, and then two-photon microscopy—to better define the measurement volume and reject background—greatly improved the signal-to-noise ratio and allowed single molecule sensitivity.[7][8] Since then, there has been a renewed interest in FCS, and as of August 2007 there have been over 3,000 papers using FCS found in Web of Science. See Krichevsky and Bonnet[9] for a recent review. In addition, there has been a flurry of activity extending FCS in various ways, for instance to laser scanning and spinning-disk confocal microscopy (from a stationary, single point measurement), in using cross-correlation (FCCS) between two fluorescent channels instead of autocorrelation, and in using Förster Resonance Energy Transfer (FRET) instead of fluorescence.
Typical FCS setupEdit
The typical FCS setup consists of a laser line (wavelengths ranging typically from 405–633 nm (cw), and from 690–1100 nm (pulsed)), which is reflected into a microscope objective by a dichroic mirror. The laser beam is focused in the sample, which contains fluorescent particles (molecules) in such high dilution, that only a few are within the focal spot (usually 1–100 molecules in one fL). When the particles cross the focal volume, they fluoresce. This light is collected by the same objective and, because it is red-shifted with respect to the excitation light it passes the dichroic mirror reaching a detector, typically a photomultiplier tube or avalanche photodiode detector. The resulting electronic signal can be stored either directly as an intensity versus time trace to be analyzed at a later point, or computed to generate the autocorrelation directly (which requires special acquisition cards). The FCS curve by itself only represents a time-spectrum. Conclusions on physical phenomena have to be extracted from there with appropriate models. The parameters of interest are found after fitting the autocorrelation curve to modeled functional forms. [10]
The measurement volumeEdit
The measurement volume is a convolution of illumination (excitation) and detection geometries, which result from the optical elements involved. The resulting volume is described mathematically by the point spread function (or PSF), it is essentially the image of a point source. The PSF is often described as an ellipsoid (with unsharp boundaries) of few hundred nanometers in focus diameter, and almost one micrometre along the optical axis. The shape varies significantly (and has a large impact on the resulting FCS curves) depending on the quality of the optical elements (it is crucial to avoid astigmatism and to check the real shape of the PSF on the instrument). In the case of confocal microscopy, and for small pinholes (around one Airy unit), the PSF is well approximated by Gaussians:
where is the peak intensity, r and z are radial and axial position, and and are the radial and axial radii, and . This Gaussian form is assumed in deriving the functional form of the autocorrelation.
Typically is 200–300 nm, and is 2–6 times larger.[11] One common way of calibrating the measurement volume parameters is to perform FCS on a species with known diffusion coefficient and concentration (see below). Diffusion coefficients for common fluorophores in water are given in a later section.
The Gaussian approximation works to varying degrees depending on the optical details, and corrections can sometimes be applied to offset the errors in approximation.[12]
Autocorrelation functionEdit
The (temporal) autocorrelation function is the correlation of a time series with itself shifted by time , as a function of :
where is the deviation from the mean intensity. The normalization (denominator) here is the most commonly used for FCS, because then the correlation at , G(0), is related to the average number of particles in the measurement volume.
Interpreting the autocorrelation functionEdit
To extract quantities of interest, the autocorrelation data can be fitted, typically using a nonlinear least squares algorithm. The fit's functional form depends on the type of dynamics (and the optical geometry in question).
Normal diffusionEdit
The fluorescent particles used in FCS are small and thus experience thermal motions in solution. The simplest FCS experiment is thus normal 3D diffusion, for which the autocorrelation is:
where is the ratio of axial to radial radii of the measurement volume, and is the characteristic residence time. This form was derived assuming a Gaussian measurement volume. Typically, the fit would have three free parameters--G(0), , and --from which the diffusion coefficient and fluorophore concentration can be obtained.
With the normalization used in the previous section, G(0) gives the mean number of diffusers in the volume <N>, or equivalently—with knowledge of the observation volume size—the mean concentration:
where the effective volume is found from integrating the Gaussian form of the measurement volume and is given by:
- gives the diffusion coefficient:
Anomalous diffusionEdit
If the diffusing particles are hindered by obstacles or pushed by a force (molecular motors, flow, etc.) the dynamics is often not sufficiently well-described by the normal diffusion model, where the mean squared displacement (MSD) grows linearly with time. Instead the diffusion may be better described as anomalous diffusion, where the temporal dependenc of the MSD is non-linear as in the power-law:
where is an anomalous diffusion coefficient. "Anomalous diffusion" commonly refers only to this very generic model, and not the many other possibilities that might be described as anomalous. Also, a power law is, in a strict sense, the expected form only for a narrow range of rigorously defined systems, for instance when the distribution of obstacles is fractal. Nonetheless a power law can be a useful approximation for a wider range of systems.
The FCS autocorrelation function for anomalous diffusion is:
where the anomalous exponent is the same as above, and becomes a free parameter in the fitting.
Using FCS, the anomalous exponent has been shown to be an indication of the degree of molecular crowding (it is less than one and smaller for greater degrees of crowding)[13].
Polydisperse diffusionEdit
If there are diffusing particles with different sizes (diffusion coefficients), it is common to fit to a function that is the sum of single component forms:
where the sum is over the number different sizes of particle, indexed by i, and gives the weighting, which is related to the quantum yield and concentration of each type. This introduces new parameters, which makes the fitting more difficult as a higher dimensional space must be searched. Nonlinear least square fitting typically becomes unstable with even a small number of s. A more robust fitting scheme, especially useful for polydisperse samples, is the Maximum Entropy Method[14].
Diffusion with flowEdit
With diffusion together with a uniform flow with velocity in the lateral direction, the autocorrelation is[15]:
where is the average residence time if there is only a flow (no diffusion).
Chemical relaxationEdit
A wide range of possible FCS experiments involve chemical reactions that continually fluctuate from equilibrium because of thermal motions (and then "relax"). In contrast to diffusion, which is also a relaxation process, the fluctuations cause changes between states of different energies. One very simple system showing chemical relaxation would be a stationary binding site in the measurement volume, where particles only produce signal when bound (e.g. by FRET, or if the diffusion time is much faster than the sampling interval). In this case the autocorrelation is:
is the relaxation time and depends on the reaction kinetics (on and off rates), and:
is related to the equilibrium constant K.
Most systems with chemical relaxation also show measureable diffusion as well, and the autocorrelation function will depend on the details of the system. If the diffusion and chemical reaction are decoupled, the combined autocorrelation is the product of the chemical and diffusive autocorrelations.
Triplet state correctionEdit
The autocorrelations above assume that the fluctuations are not due to changes in the fluorescent properties of the particles. However, for the majority of (bio)organic fluorophores--e.g. green fluorescent protein, rhodamine, Cy3 and Alexa Fluor dyes--some fraction of illuminated particles are excited to a triplet state (or other non-radiative decaying states) and then do not emit photons for a characteristic relaxation time . Typically is on the order of microseconds, which is usually smaller than the dynamics of interest (e.g. ) but large enough to be measured. A multiplicative term is added to the autocorrelation account for the triplet state. For normal diffusion:
where is the fraction of particles that have entered the triplet state and is the corresponding triplet state relaxation time. If the dynamics of interest are much slower than the triplet state relaxation, the short time component of the autocorrelation can simply be truncated and the triplet term is unnecessary.
Common fluorescent probesEdit
The fluorescent species used in FCS is typically a biomolecule of interest that has been tagged with a fluorophore (using immunohistochemistry for instance), or is a naked fluorophore that is used to probe some environment of interest (e.g. the cytoskeleton of a cell). The following table gives diffusion coefficients of some common fluorophores in water at room temperature, and their excitation wavelengths.
|Fluorescent dye
|(×10−10 m2 s−1)
|Excitation wavelength (nm)
|Rhodamine 6G
|[16], [17], [18]
|Rhodamine 110
|Tetramethyl rhodamine
|[20], [21]
|2′, 7′-difluorofluorescein (Oregon Green488)
Variations of FCSEdit
FCS almost always refers to the single point, single channel, temporal autocorrelation measurement, although the term "fluorescence correlation spectroscopy" out of its historical scientific context implies no such restriction. FCS has been extended in a number of variations by different researchers, with each extension generating another name (usually an acronym).
Fluorescence cross-correlation spectroscopy (FCCS)Edit
FCS is sometimes used to study molecular interactions using differences in diffusion times (e.g. the product of an association reaction will be larger and thus have larger diffusion times than the reactants individually); however, FCS is relatively insensitive to molecular mass as can be seen from the following equation relating molecular mass to the diffusion time of globular particles (e.g. proteins):
where is the viscosity of the sample and is the molecular mass of the fluorescent species. In practice, the diffusion times need to be sufficiently different--a factor of at least 1.6--which means the molecular masses must differ by a factor of 4.[26] Dual color fluorescence cross-correlation spectroscopy (FCCS) measures interactions by cross-correlating two or more fluorescent channels (one channel for each reactant), which distinguishes interactions more sensitively than FCS, particularly when the mass change in the reaction is small.
Brightness analysis methods (N&B,[27] PCH,[28] FIDA,[29] Cumulant Analysis[30])Edit
Fluorescence cross correlation spectroscopy overcomes the weak dependence of diffusion rate on molecular mass by looking at multicolor coincidence. What about homo-interactions? The solution lies in brightness analysis. These methods use the heterogeneity in the intensity distribution of fluorescence to measure the molecular brightness of different species in a sample. Since dimers will contain twice the number of fluorescent labels as monomers, their molecular brightness will be approximately double that of monomers. As a result, the relative brightness is sensitive a measure of oligomerization. The average molecular brightness ( ) is related to the variance ( ) and the average intensity ( ) as follows:[31]
Here and are the fractional intensity and molecular brightness, respectively, of species .
Two- and three- photon FCS excitationEdit
Several advantages in both spatial resolution and minimizing photodamage/photobleaching in organic and/or biological samples are obtained by two-photon or three-photon excitation FCS[32][33][34][35][36].
Another FCS based approach to studying molecular interactions uses fluorescence resonance energy transfer (FRET) instead of fluorescence, and is called FRET-FCS.[37] With FRET, there are two types of probes, as with FCCS; however, there is only one channel and light is only detected when the two probes are very close—close enough to ensure an interaction. The FRET signal is weaker than with fluorescence, but has the advantage that there is only signal during a reaction (aside from autofluorescence).
Image correlation spectroscopy (ICS)Edit
When the motion is slow (in biology, for example, diffusion in a membrane), getting adequate statistics from a single-point FCS experiment may take a prohibitively long time. More data can be obtained by performing the experiment in multiple spatial points in parallel, using a laser scanning confocal microscope. This approach has been called Image Correlation Spectroscopy (ICS)[38]. The measurements can then be averaged together.
Another variation of ICS performs a spatial autocorrelation on images, which gives information about the concentration of particles[39]. The correlation is then averaged in time.
A natural extension of the temporal and spatial correlation versions is spatio-temporal ICS (STICS) [40]. In STICS there is no explicit averaging in space or time (only the averaging inherent in correlation). In systems with non-isotropic motion (e.g. directed flow, asymmetric diffusion), STICS can extract the directional information. A variation that is closely related to STICS (by the Fourier transform) is k-space Image Correlation Spectroscopy (kICS).[41]
There are cross-correlation versions of ICS as well.[38]
Scanning FCS variationsEdit
Some variations of FCS are only applicable to serial scanning laser microscopes. Image Correlation Spectroscopy and its variations all were implemented on a scanning confocal or scanning two photon microscope, but transfer to other microscopes, like a spinning disk confocal microscope. Raster ICS (RICS)[42], and position sensitive FCS (PSFCS)[43] incorporate the time delay between parts of the image scan into the analysis. Also, low dimensional scans (e.g. a circular ring)[44]—only possible on a scanning system—can access time scales between single point and full image measurements. Scanning path has also been made to adaptively follow particles.[45]
Spinning disk FCS, and spatial mappingEdit
Any of the image correlation spectroscopy methods can also be performed on a spinning disk confocal microscope, which in practice can obtain faster imaging speeds compared to a laser scanning confocal microscope. This approach has recently been applied to diffusion in a spatially varying complex environment, producing a pixel resolution map of a diffusion coefficient.[46]. The spatial mapping of diffusion with FCS has subsequently been extended to the TIRF system.[47] Spatial mapping of dynamics using correlation techniques had been applied before, but only at sparse points[48] or at coarse resolution[40].
Total internal reflection FCSEdit
Total internal reflection fluorescence (TIRF) is a microscopy approach that is only sensitive to a thin layer near the surface of a coverslip, which greatly minimizes background fluorscence. FCS has been extended to that type of microscope, and is called TIR-FCS[49]. Because the fluorescence intensity in TIRF falls off exponentially with distance from the coverslip (instead of as a Gaussian with a confocal), the autocorrelation function is different.
Other fluorescent dynamical approachesEdit
There are two main non-correlation alternatives to FCS that are widely used to study the dynamics of fluorescent species.
Fluorescence recovery after photobleaching (FRAP)Edit
In FRAP, a region is briefly exposed to intense light, irrecoverably photobleaching fluorophores, and the fluorescence recovery due to diffusion of nearby (non-bleached) fluorophores is imaged. A primary advantage of FRAP over FCS is the ease of interpreting qualitative experiments common in cell biology. Differences between cell lines, or regions of a cell, or before and after application of drug, can often be characterized by simple inspection of movies. FCS experiments require a level of processing and are more sensitive to potentially confounding influences like: rotational diffusion, vibrations, photobleaching, dependence on illumination and fluorescence color, inadequate statistics, etc. It is much easier to change the measurement volume in FRAP, which allows greater control. In practice, the volumes are typically larger than in FCS. While FRAP experiments are typically more qualitative, some researchers are studying FRAP quantitatively and including binding dynamics.[50] A disadvantage of FRAP in cell biology is the free radical perturbation of the cell caused by the photobleaching. It is also less versatile, as it cannot measure concentration or rotational diffusion, or co-localization. FRAP requires a significantly higher concentration of fluorophores than FCS.
Particle trackingEdit
In particle tracking, the trajectories of a set of particles are measured, typically by applying particle tracking algorithms to movies.[1] Particle tracking has the advantage that all the dynamical information is maintained in the measurement, unlike FCS where correlation averages the dynamics to a single smooth curve. The advantage is apparent in systems showing complex diffusion, where directly computing the mean squared displacement allows straightforward comparison to normal or power law diffusion. To apply particle tracking, the particles have to be distinguishable and thus at lower concentration than required of FCS. Also, particle tracking is more sensitive to noise, which can sometimes affect the results unpredictably.
- ↑ Magde, D., Elson, E. L., Webb, W. W. Thermodynamic fluctuations in a reacting system: Measurement by fluorescence correlation spectroscopy,(1972) Phys Rev Lett, 29, 705–708.
- ↑ Ehrenberg, M., Rigler, R. Rotational brownian motion and fluorescence intensity fluctuations,(1974) Chem Phys, 4, 390–401.
- ↑ Elson, E. L., Magde, D. Fluorescence correlation spectroscopy I. Conceptual basis and theory,(1974) Biopolymers, 13, 1–27.
- ↑ Magde, D., Elson, E. L., Webb, W. W. Fluorescence correlation spectroscopy II. An experimental realization,(1974) Biopolymers, 13, 29–61.
- ↑ Thompson N L 1991 Topics in Fluorescence Spectroscopy Techniques vol 1, ed J R Lakowicz (New York: Plenum) pp 337–78
- ↑ Rigler, R, Ü. Mets1, J. Widengren and P. Kask. Fluorescence correlation spectroscopy with high count rate and low background: analysis of translational diffusion. European Biophysics Journal (1993) 22(3), 159.
- ↑ Eigen, M., Rigler, M. Sorting single molecules: application to diagnostics and evolutionary biotechnology,(1994) Proc. Natl. Acad. Sci. USA, 91,5740–5747.
- ↑ Rigler, M. Fluorescence correlations, single molecule detection and large number screening. Applications in biotechnology,(1995) J. Biotechnol., 41,177–186.
- ↑ O. Krichevsky, G. Bonnet, "Fluorescence correlation spectroscopy: the technique and its applications," Rep. Prog. Phys. 65, 251–297 (2002).
- ↑ Medina, M. A., Schwille, P. Fluorescence correlation spectroscopy for the detection and study of single molecules in biology, (2002)BioEssays, 24, 758–764.
- ↑ Mayboroda, O. A., van Remoortere, A., Tanke H. J., Hokke, C. H., Deelder, A. M., A new approach for fluorescence correlation spectroscopy (FCS) based immunoassays, (2003), J. Biotechnol., 107, 185–192.
- ↑ Hess, S.T., and W.W. Webb. 2002. Focal volume optics and experimental artifacts in confocal fluorescence correlation spectroscopy. Biophys. J. 83:2300–2317.
- ↑ Banks, D. S., and C. Fradin. 2005. Anomalous diffusion of proteins due to molecular crowding. Biophys. J. 89:2960–2971.
- ↑ Sengupta, P., K. Garai, J. Balaji, N. Periasamy, and S. Maiti. 2003. Measuring Size Distribution in Highly Heterogeneous Systems with Fluorescence Correlation Spectroscopy. Biophys. J. 84(3):1977–1984.
- ↑ Kohler, R.H., P. Schwille, W.W. Webb, and M.R. Hanson. 2000. Active protein transport through plastid tubules: velocity quantified by fluorescence correlation spectroscopy. J Cell Sci 113(22):3921–3930
- ↑ Magde, D., Elson, E. L., Webb, W. W. Fluorescence correlation spectroscopy II. An experimental realization,(1974) Biopolymers, 13,29–61.
- ↑ Berland, K. M. Detection of specific DNA sequences using dual-color two-photon fluorescence correlation spectroscopy. (2004),J. Biotechnol ,108(2), 127–136.
- ↑ Müller, C.B., Loman, A., Pacheco, V., Koberling, F., Willbold, D., Richtering, W., Enderlein, J. Precise measurement of diffusion by multi-color dual-focus fluorescence correlation spectroscopy (2008), EPL, 83, 46001.
- ↑ Pristinski, D., Kozlovskaya, V., Sukhishvili, S. A. Fluorescence correlation spectroscopy studies of diffusion of a weak polyelectrolyte in aqueous solutions. (2005), J. Chem. Phys., 122, 014907.
- ↑ Widengren, J., Schwille, P., Characterization of photoinduced isomerization and back-isomerization of the cyanine dye Cy5 by fluorescence correlation spectroscopy. (2000), J. Phys. Chem. A, 104, 6416–6428.
- ↑ Loman, A., Dertinger, T., Koberling, F., Enderlein, J. Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy (2008), Chem. Phys. Lett., 459, 18–21.
- ↑ Pristinski, D., Kozlovskaya, V., Sukhishvili, S. A. Fluorescence correlation spectroscopy studies of diffusion of a weak polyelectrolyte in aqueous solutions. (2005), J. Chem. Phys., 122, 014907.
- ↑ Müller, C.B., Loman, A., Pacheco, V., Koberling, F., Willbold, D., Richtering, W., Enderlein, J. Precise measurement of diffusion by multi-color dual-focus fluorescence correlation spectroscopy (2008), EPL, 83, 46001.
- ↑ Müller, C.B., Loman, A., Pacheco, V., Koberling, F., Willbold, D., Richtering, W., Enderlein, J. Precise measurement of diffusion by multi-color dual-focus fluorescence correlation spectroscopy (2008), EPL, 83, 46001.
- ↑ Müller, C.B., Loman, A., Pacheco, V., Koberling, F., Willbold, D., Richtering, W., Enderlein, J. Precise measurement of diffusion by multi-color dual-focus fluorescence correlation spectroscopy (2008), EPL, 83, 46001.
- ↑ Meseth, U., Wohland, T., Rigler, R., Vogel, H. Resolution of fluorescence correlation measurements. (1999) Biophys. J., 76, 1619–1631.
- ↑ Digman, M. A., R. Dalal, A. F. Horwitz, and E. Gratton. Mapping the number of molecules and brightness in the laser scanning microscope. (2008) Biophys. J. 94, 2320–2332.
- ↑ Chen, Y., J. D. Müller, P. T. C. So, and E. Gratton. The photon counting histogram in fluorescence fluctuation spectroscopy. (1999) Biophys. J. 77, 553–567.
- ↑ Kask, P., K. Palo, D. Ullmann, and K. Gall. Fluorescence-intensity distribution analysis and its application in biomolecular detection technology. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 13756–13761.
- ↑ Müller, J. D. Cumulant analysis in fluorescence fluctuation spectroscopy. (2004) Biophys. J. 86, 3981–3992.
- ↑ Qian, H., Elson, E.L. On the analysis of high order moments of fluorescence fluctuations. (1990) Biophys. J., 57, 375–380.
- ↑ Diaspro, A., and Robello, M. (1999). Multi-photon Excitation Microscopy to Study Biosystems. European Microscopy and Analysis., 5:5–7.
- ↑ Bagatolli, L.A., and Gratton, E. (2000). Two-photon fluorescence microscopy of coexisting lipid domains in giant unilamellar vesicles of binary phospholipid mixtures. Biophys J., 78:290–305.
- ↑ Schwille, P., Haupts, U., Maiti, S., and Webb. W.(1999). Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two- photon excitation. Biophysical Journal, 77(10):2251–2265.
- ↑ Near Infrared Microspectroscopy, Fluorescence Microspectroscopy,Infrared Chemical Imaging and High Resolution Nuclear Magnetic Resonance Analysis of Soybean Seeds, Somatic Embryos and Single Cells., Baianu, I.C. et al. 2004., In Oil Extraction and Analysis., D. Luthria, Editor pp.241–273, AOCS Press., Champaign, IL.
- ↑ Single Cancer Cell Detection by Near Infrared Microspectroscopy, Infrared Chemical Imaging and Fluorescence Microspectroscopy.2004.I. C. Baianu, D. Costescu, N. E. Hofmann and S. S. Korban, q-bio/0407006 (July 2004)
- ↑ K. Remaut, B. Lucas, K. Braeckmans, N.N. Sanders, S.C. De Smedt and J. Demeester, FRET-FCS as a tool to evaluate the stability of oligonucleotide drugs after intracellular delivery, J Control Rel 103 (2005) (1), pp. 259–271.
- ↑ a b Wiseman, P. W., J. A. Squier, M. H. Ellisman, and K. R. Wilson. 2000. Two-photon video rate image correlation spectroscopy (ICS) and image cross-correlation spectroscopy (ICCS). J. Microsc. 200:14–25.
- ↑ Petersen, N. O., P. L. Ho¨ddelius, P. W. Wiseman, O. Seger, and K. E. Magnusson. 1993. Quantitation of membrane receptor distributions by image correlation spectroscopy: concept and application. Biophys. J. 65:1135–1146.
- ↑ a b Hebert, B., S. Constantino, and P. W. Wiseman. 2005. Spatio-temporal image correlation spectroscopy (STICS): theory, verification and application to protein velocity mapping in living CHO cells. Biophys. J. 88:3601–3614.
- ↑ Kolin, D.L., D. Ronis, and P.W. Wiseman. 2006. k-Space Image Correlation Spectroscopy: A Method for Accurate Transport Measurements Independent of Fluorophore Photophysics. Biophys. J. 91(8):3061–3075.
- ↑ Digman, M.A., P. Sengupta, P.W. Wiseman, C.M. Brown, A.R. Horwitz, and E. Gratton. 2005. Fluctuation Correlation Spectroscopy with a Laser-Scanning Microscope: Exploiting the Hidden Time Structure. Biophys. J. 88(5):L33–36.
- ↑ Skinner, J.P., Y. Chen, and J.D. Mueller. 2005. Position-Sensitive Scanning Fluorescence Correlation Spectroscopy. Biophys. J.:biophysj.105.060749.
- ↑ Ruan, Q., M.A. Cheng, M. Levi, E. Gratton, and W.W. Mantulin. 2004. Spatial-temporal studies of membrane dynamics: scanning fluorescence correlation spectroscopy (SFCS). Biophys. J. 87:1260–1267.
- ↑ A. Berglund and H. Mabuchi, "Tracking-FCS: Fluorescence correlation spectroscopy of individual particles," Opt. Express 13, 8069–8082 (2005).
- ↑ Sisan, D.R., R. Arevalo, C. Graves, R. McAllister, and J.S. Urbach. 2006. Spatially resolved fluorescence correlation spectroscopy using a spinning disk confocal microscope. Biophysical Journal 91(11):4241–4252.
- ↑ Kannan, B., L. Guo, T. Sudhaharan, S. Ahmed, I. Maruyama, and T. Wohland. 2007. Spatially resolved total internal reflection fluorescence correlation microscopy using an electron multiplying charge-coupled device camera. Analytical Chemistry 79(12):4463–4470
- ↑ Wachsmuth, M., W. Waldeck, and J. Langowski. 2000. Anomalous diffusion of fluorescent probes inside living cell nuclei investigated by spatially-resolved fluorescence correlation spectroscopy. J. Mol. Biol. 298(4):677–689.
- ↑ Lieto, A.M., and N.L. Thompson. 2004. Total Internal Reflection with Fluorescence Correlation Spectroscopy: Nonfluorescent Competitors. Biophys. J. 87(2):1268–1278.
- ↑ Sprague, B.L., and J.G. McNally. 2005. FRAP analysis of binding: proper and fitting. Trends in Cell Biology 15(2):84–91.
Further readingEdit
- Rigler R. and Widengren J. (1990). Ultrasensitive detection of single molecules by fluorescence correlation spectroscopy, BioScience (Ed. Klinge & Owman) p.180
- Oehlenschläger F., Schwille P. and Eigen M. (1996). Detection of HIV-1 RNA by nucleic acid sequence-based amplification combined with fluorescence correlation spectroscopy, Proc. Natl. Acad. Sci. USA 93:1281.


text:
Life gets the knowledge of life through teachers. The lesson of living life is taught by the teachers.
The one who puts the children of the society in the right path of life. Though they themselves spend simple and decent life, playing a key role in enabling the country.
Holding the same status as our parents, they teach the right way to live even in difficult times of life.
And the ultimate Backbone for the future Healthy Nation.
Teachers are the one who influences the lives of children to a great extent. Lessons learned by children in schools remain with them for a long time. Teachers are often known as creator of future of nation too. Like every year, the day to wish teachers is nearby.
World teacher's day is celebrated on 5th of October every year. Every country celebrates teacher's day to recognize and appreciate teachers. In different countries the day is decided according to the special day of their local educationist. The day and the way of celebrating teacher's day may be different. But the aim is same, to show gratitude towards teacher who construct the future of every nation through their knowledge.
Related Post: Top 10 Time Management Tips For Students.
In India, Teacher's day is observed on 5th September. It is celebrated to applause the contribution of Dr. Sarvepalli Radhakrishnan in the field of educations. He was born on September 5, 1888. He was a great scholar, philosopher, a Bharat Ratna recipient, first vice president and second president of India. But before anything else, He was a teacher. He was a very friendly teacher and quite popular among his students for the examples he sets in front of them.
One day his students shown the will to celebrate his birthday in a lavish way. He accepted their request and said that it would be his proud privilege, if my birthday is celebrated to give respect to all the teachers. So, since 1962 when he started serving as second President of India, we are celebrating his birth anniversary as Teacher's day to honor his work.
He also wrote a book "Political thinkers of Modern India", in which he signified about the importance of teachers and education in a country. In his words, teachers should be respected more because they play a huge role in building the nation. They are the architect of society. Once he wrote Bhagwad Geeta and there he defined teachers as, "The one who emphasizes on presentation to converge different currents of thoughts to the same end". Apart from a teacher and a thinker, he was a great philosopher. When he entered into the politics, leaders like Mahatma Gandhi, Jawaharlal Nehru, Dr. Rajendra Prasad were already the fan of his thinking of nation building. Such a great man truly deserves a standing ovation for his marvelous thinking and endeavors.
The teacher is a guide, philosopher, friend who impart us the important lessons of life. He holds our hand to guide us, open our mind through his wisdom and touches our heart with his knowledge. We can't ignore the contribution of teacher at all. They give shape to our mind through their knowledge and make us a responsible citizen.
That's why this day is celebrated around the world to applaud teachers. While in schools, colleges teachers are honored on this special day for their work. In many schools, on Teacher's day student plays the part of teachers and give the lectures. Teachers sit in classes like students and revive their student life.

subset: commoncrawl


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Car Insurance for Mexico from MexicanInsuranceStore.com – Don't Be Forgetful
Forgetfulness with a big trip isn't unusual, but it can have some serious consequences if you forget the wrong thing. Planning for a few days in Mexico involves some documents and details that you can't afford to be without. One of those details is your car insurance for Mexico from MexicanInsuranceStore.com. You can deal with buying Mexican insurance online at MexicanInsuranceStore.com well in advance of your travels so that you can forget about it, but make sure that your policy documents are printed and safely stowed in your vehicle.
Will Anyone Check for My Car Insurance for Mexico from MexicanInsuranceStore.com?
If you drive without any significant incidents during your travels, it is unlikely that any official check of your coverage documents will be made. However, you should always be prepared for the unexpected in Mexico. Military checkpoints, for example, can vary from day to day. You may encounter special units at times, and some officials will ask about tourist visas, Mexican insurance online at MexicanInsuranceStore.com and passports. If you are stopped by a police officer for an infraction, you may be more likely to need to produce your Car Insurance for Mexico from MexicanInsuranceStore.com. It's cheap car insurance that anyone with a bicycle can afford, so there's not much harm in keeping paperwork like that updated.
If you are in an accident, you will definitely need to produce your documents. These documents serve as proof of ability to shoulder financial responsibility for damages. It's always smart to make an extra copy of the documents in case of theft or any incident with your vehicle when you are away from it.
What If I'm in Mexico without Car Insurance for Mexico from MexicanInsuranceStore.com?
If you've made it across the border before remembering to buy your coverage, you will find that it's very easy to handle it right away. You can look up and buy Mexican insurance online at MexicanInsuranceStore.com with your wireless phone or another mobile device. You will need to enter the specifics about your model and its value. You need to decide about the type of coverage and the limits you need for your trip. You can complete the transaction quickly, and your coverage will be transmitted right away to Mexican databases, meaning that your coverage can be verified if you have an accident before you can print a copy of your policy.
Car Insurance for Mexico from MexicanInsuranceStore.com comes with FREE Roadside Assistance
Cancun and the Mexican Caribbean Weather


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It may have started as an exclusive network for college students, but the transition has been completed: Facebook is for old people.
According to a report from eMarketer, 2.2 million 12- to 17-year-olds and 4.5 million 18- to 24-year-olds will use Facebook in the UK this year. That figure is 700,00 fewer than in 2017.
This drop, along with an influx of older users, means that over-55s will become the second-largest demographic of Facebook users this year.
500,000 new over-55s are expected to join Facebook this year. There will be 6.4 million 55- to 65-year-old-plus regular Facebook users this year, the biggest demographic save for 16- to 34-year-olds.
"There are a couple of factors at play here," noted Ampere analyst Richard Broughton. "One is that older people tend to be late to the internet party, but adoption tends to find its way through the demographics eventually. And with Facebook's video and photo experience it is a platform they want to be on to keep up with the social lives of their kids and grandchildren."
However it's not all doom and gloom as far as Facebook's youth appeal goes. While younger users may be fleeing from the main Facebook brand, they are largely shifting to services such as Instagram and WhatsApp which are owned by Facebook. Some though are entirely abandoning Facebook for rival services like Snapchat and Twitter.
eMarketer UK senior analyst Bill Fisher was very matter of fact: "Facebook has a teen problem.
"This latest forecast indicates that it is more than just a theory. Until now it has been able to rely on platform shifters being hoovered up by Instagram. However, leading the charge for younger audiences is Snapchat. There are now some early signs that younger social network users are being swayed by Snapchat."
According to Fisher, Snapchat's user-ship has more than doubled among UK social networking users to 43 per cent.
Facebook UK users 2018 v 2017
Age 12 to 17 2.2m, down 300,000
Age 18 to 24 4.5m, down 400,000
Age 25 to 34 7.2m, flat
Age 35 to 44 5.9m, flat
Age 45 to 54 5.6m, up 100,000 users
Age 55 to 64 3.5m, up 200,000 users
Age 65-plus 2.9m, up 300,000 users


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Ithaca's cooperatively-owned independent bookstore since 2011
215 N Cayuga Street Dewitt Mall · Ithaca, NY · 607-273-8246
It is easy to forget that every development in the history of the American information industry--from the telephone to radio to film--once existed in an open and chaotic marketplace inhabited by entrepreneurs and utopians, just as the Internet does today. Each of these, however, grew to be dominated by a monopolist or cartel. In this path-breaking audiobook, Tim Wu asks: will the Internet follow the same fate? Could the Web--the entire flow of American information--come to be ruled by a corporate leviathan in possession of "the master switch"?
Analyzing the strategic maneuvers of today's great information powers--Apple, Google, and an eerily resurgent AT&T--Wu uncovers a time-honored pattern in which invention begets industry and industry begets empire. He shows how a battle royal for the Internet's future is brewing, and this is one war we dare not tune out.
"Fascinating, balanced, and rigorous--a tour de force." --The New York Review of Books
"Entertaining.... There's a sharp insight and a surprising fact on nearly every page of Wu's masterful survey." --The Boston Globe


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|Showing 1 of 1
Click on a column heading to sort the list.
|Medium & Support
|Creation Date
|Farm Scene, after Saftleven (?)
|Ploos van Amstel, Cornelis
|Pen and black ink with brush and brown wash over graphite
|circa 1765
Your current search criteria is: Keyword is "I" and [Object]Artist is "Cornelis Ploos van Amstel".View current selection of records as:


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DELAC Meeting
Our District English Learner Advisory Committee (DELAC) will be hosting meetings in the Annex (Board Side) at the Administration & Educational Support Center.
DELAC is a district-level English Learner Advisory Committee comprised of parents, staff, and community members designated to advise district officials on English learner programs and services.
Parents of English Learner students can attend in order to provide input on the committee's recommendations.
DATE: February 7, 2019
TIME: 11:00 a.m.
Hesperia Unified School District
Annex Building, Board Side
15576 Main Street
Hesperia, CA 92345
- Opening
- Roll Call
- Topics - California School Dashboard
- Questions & Comments
- Meeting Adjournment
Join us, get involved, get informed ...
Let's work together for the benefit of our children.


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Many experts believed driving was too complex to be automated, yet here we are with self driving taxis, cars, and trucks. The robots are coming for drivers - and they are coming for you too. No matter what your job, a robot WILL do it. They will be stronger, faster, and smarter than you. They will be better writers, better doctors, and run companies more effectively.
We can delay the inevitable by shutting down borders and subsidizing losing industries, but we will not stop robots from replacing human labor (nor should we). The challenge is in the transition from a human labor to the robot run economy. Many people will struggle to find new places for their skills, and the result could be increased income inequality, increased violence, and an uprooted sense of self.
In this talk, we'll explore our responsibility as technologists to be aware of and address this inevitable transition to a robot labor force.


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Walter Small Business: For years we had a great business relationship. We shared rental space, did joint promotions, and made money together... not a lot of money, but we made money. Then he got into trouble with the IRS, something about back taxes on some land. He was in a bind so I loaned him some money.
Gene Skiptracer: How much money?
WSB: $8,700.00! That was three years ago. When he got back on his feet again, he wouldn't talk to me about repaying my money, or even return my phone calls. So I went to court and got a judgment against him.
GS: What type of business does he have?
WSB: He owns PR Trucking Company. He has five trucks and his biggest customer is Stay Lowe Food Market
GS: Is your judgment against him or against the company?
WSB: Against both.
GS: I think you can get your money fairly easy. Who supplies his gas?
WSB: Reader Oil Company. He pays for consignments and has a storage tank almost as big as a gas station. They make two deliveries a month.
GS: This is how you can get his attention and make him pay. Have your attorney file garnishment papers requesting a third party levy. A third party levy is a powerful collection tool.. You can garnish the Stay Lowe Food Store and request that it surrenders all money being held for the judgment debtor. You can also use a third party levy to seize all products being held by a third party for the judgment debtor. If PR Trucking pays Reader Oil Company for a consignment of gas, you can petition the court to order Reader Oil Company to release the gas to you. Once it's in your possession you can auction it off to pay for your judgment. Third party levies can be used to garnish bank accounts, stocks and bonds, or any other valuables being held by a third party.


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Louise tells the group she hates her body and she has had spinal surgery for scoliosis.
I tell her that she may have received a dose of hate while she was still in the womb. Her body is wounded, but not ugly.
KBW "Your psychological wound seems to have been created by an early dose of hate, usually in the first 3 months of life. You may have felt you were not lovable and not wanted. At times you may have felt something like 'what's the use of being here; why even go on'. This is not a suicidal thought, but a sense of not belonging, not fitting in, in any shape or form. Some people who receive this wound are very spiritual. Are you spiritual? "
L "Yes, I am."
"KBW "You also tend to detach from what is happening around you. Do you tune out some of the time?"
L "Yes, I do."
KBW "Your chest is relatively boney and the ribs stick out. This is generally a sign of early deprivation and abandonment. Your parenting figures may not have supported you, although some of this may have happened in a past life."
"The collapse in your feet at the arch shows feelings of unworthiness and not being able to stand up for yourself. "
L "My defense is to prove the opposite, that I am worthy."
KBW "You stand with your feet fairly wide apart in an effort to ground and make up for the support you did not receive. The problem is that by standing with your legs and feet far apart your life energy is free to dissipate, so that it is no longer yours."
KBW "The scoliosis in your spine shows that when you came into this world you brought with you a powerful soul or personality. You were more powerful than your family could handle, and in order to live in your family you had to squash this power. When the child squashes this power it distorts the spine and leads to scoliosis."
KBW "You must be a very strong person, for even with this kind of wounding, you seem to function well. You did not end up in a mental hospital."
KBW "I suggest to you work on your anger (when the young child or infant receives hate, she mirrors it back to the world in the form of suppressed anger or hate.) Once you release the anger, you are free to develop your femininity and your soft side. Your body has not yet filled in the feminine softness. Your legs are positioned apart in a masculine stance, and by keeping them so far apart you lose your grounding and your sexual energy."
KBW "What is your purpose here on the planet? How can you achieve more personal fulfillment. With this wound, it is important to explore your life purpose and to make a full commitment to life."
L "I exist and I fight back. I am a life coach and I am active in politics."
KBW "What do you want for yourself? "
L "I want love, but I don't know how to get it."
KBW "You must open your heart, but that goes hand and hand with getting out your anger."
We end when I ask her to open her arms toward the group of people who are observing the reading and embrace them and receive their loving feelings. Many of whom come up and hug her.


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Thursday, July 17, 2014
Getting active again
I plan to play golf with friends. Also, I take golf lessons with a group from the local junior college at a nearby driving range. I have a good friend who has a golf simulator in his garage and I play simulated golf on his system about once each week. His simulator has golf courses from all over the United States in memory.
On my apartment patio, I am growing a bunch of flowering plants. They are all in bloom now.
I've got a big tub on the balcony which is serving as a pond. There are several pond plants growing now. One of them is a flowering lily. No flowers yet from this lily, but several new leaves have sprouted! I have hopes for flowers from this lily plant.
I do nearly all of my shopping by driving on my electric bike to the market. I use a backpack to carry what I buy back to the apartment. Since I am not able to carry too much in that backpack, I go shopping nearly every day.
I workout at the YMCA about three times each week. Very important for me to exercise, since I have been diagnosed with osteoporosis. I am taking a lot of vitamins which is supposed to curb osteoporosis. Vitamin D3 and calcium in very large doses.
I am also beginning to do activities at a senior citizen center which is close to my apartment.
14 comments:
Glad your foot is better & you are able to get back to your fun activities.ReplyDelete
Stay well.
As you know, staying active slows the effect of the years.ReplyDelete
George, I'm happy to hear you're living life! Praise God! Prayer for you, continues...ReplyDelete
Your an inspiration to all of us George. Keep er goin:))ReplyDelete
George, after all your hard work setting up the garden and fountain I am so glad you have flowers, leaves, and progress...... Hope the water lily produces a beautiful bunch of blooms !!!! You are so amazing and as Al said such an inspiration !!!ReplyDelete
- Anonymous7/17/14, 11:36 PM
Good for you George. I hope you stay healthy and active.ReplyDelete
I have concerns when you say you are taking large amounts of calcium. I hope it is a decision with your doctor. There was a recent report on large doses of calcium and heart problems. One does. to know what to believe anymore. BTW, my husband just got back from his doctor because of his foot pain, and he was diagnosed with gout.ReplyDeleteReplies
I am taking 5000 units Vitamin D3 in the AM and again in the PM.Delete
Calcium 600mg in the AM and again in the PM.
Sounds like you're doing good with the vitamins. We take mega-doses of Vit C, 12,000 mg plus Calcium, VitD,E, Omega, Q10, multi megas. Diet can't do it and neither can the body. Here is the scoop on Calcium.ReplyDelete
"The Institute of Medicine sets the daily tolerable upper intake level (UL) for calcium based on age as follows: Age 0-6 months, 1000 mg; 6-12 months, 1500 mg; 1-3 years, 2500 mg; 9-18 years, 3000 mg; 19-50 years, 2500 mg; 51+ years, 2000 mg. Higher doses increase the chance of having serious side effects. Some recent research also suggests that doses over the recommended daily requirement of 1000-1300 mg daily for most adults might increase the chance of heart attack."
Recently my doctor lowered my intake of calcium. He said there are risks associated with intake of large doses of calcium...but I didn't ask what....I assumed brittle bones. I will ask the next time I see him. Take care George.ReplyDelete
George—Diana and I send you our very best wishes for a full recover. I'm only now getting us caught up on all your life "events" of the past few months.ReplyDelete
Regards, Allen, Diana, (Rooney and Daphne too) Russell
Hey George! Where are you? Miss reading about your daily adventures!ReplyDelete
George - too long in between posts. Let us know how you are and what is going on.ReplyDelete
How are you, George? Long time since your last post.ReplyDelete


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Eclipse IDE
1502 Solutions and counting.
Update March 2019:
be sure to use https://nodeclipse.github.io/updates/gradle-ide-pack/
not nodeclipse.org
Install Pivotal Gradle IDE & Enide Gradle for Eclipse & EGradle Edi...
The JFrog Eclipse Plugin adds JFrog Xray scanning of Maven, Gradle and NPM project dependencies to your Eclipse IDE. It allows developers to view panels displaying vulnerability information ab...
EGradle is a lightweight, fast and comfortable gradle integration for eclipse
Features overview
Console and Eclipse can be used parallel without conflicts
New Project...
Artifactory - P2 Repositories can be your single access point for all Eclipse® updates. Eclipse plugins proxying and hosting take advantage of Artifactory's exiting advanced caching and security co...
https://github.com/Nodeclipse/GradleEditor Minimalist Gradle Editor for build.gradle files with highlight for keywords, strings and matching brackets and android support (by taking some additional ...
Nodeclipse "Enide Studio 2015" is Tool Suite for Node.js, JavaScript, Java Development. This is the most feature-rich pack. Unless there is the latest version download for your OS, recomm...
Nodeclipse "Enide Studio 2014" is Tool Suite for Node.js, JavaScript, Java Development.
This is the most feature-rich pack. Unless there is the latest version download for your OS,
recommended ...
For any project that gradle can build. Launch build, gradle GUI, run Jetty or deploy Android .apk to AVD by right-clicking build.gradle. Project does not need to be Gradle project. Just build.gradl...
IDE, Tools, Editor, Build and Deploy
Last Updated on Friday, January 15, 2021 - 10:11 by Paul Verest
EGradle Editor as standalone plugin, so other IDEs can use the editor implementation as well or users just install the editor.
The EGradle IDE does contain this editor which has much features:
Build and Deploy, Tools, Editor
Last Updated on Friday, July 15, 2022 - 10:20 by Albert Tregnaghi
Last Updated on Friday, January 15, 2021 - 10:12 by Paul Verest
Build and Deploy, IDE, Editor, Tools
Last Updated on Friday, July 15, 2022 - 10:19 by Albert Tregnaghi
Application Development Frameworks, Languages, IDE, Editor
Application Development Frameworks, IDE, Languages, Editor
Last Updated on Monday, February 4, 2019 - 13:54 by Paul Verest
Tools, Editor, Build and Deploy
Last Updated on Friday, January 15, 2021 - 10:13 by Paul Verest
Source Code Analyzer, Reporting, Tools, Code Management
Last Updated on Monday, August 16, 2021 - 07:58 by JFrog JFrog
In a nutshell:
This plugin is not a replacement for JDT JUnit view, but more an addition for junit server results. It shows server console output and error stream in own console, can also correl...
Tools, Testing
Build and Deploy
Last Updated on Monday, November 14, 2011 - 09:20 by Shlomi Ben Haim
This feature contains a bndtools workspace with an extensive Gradle integration, one that is far more flexible and featureful than what is delivered by bndtools itself. Bndtools is an Eclipse-based...
Build and Deploy, Tools
Last Updated on Wednesday, December 11, 2019 - 13:05 by Ferry Huberts


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The Audit & Risk Recruitment Company (ARRC) has been asked by a Top 10 accounting firm in central London or West Midlands to recruit for a Senior IT Auditor. You will be reporting to the senior management and can expect to work on top public sector clients. There is low travel with this role, mainly within the Midlands area.
You will be joining a dynamic team who are leaders in providing IT internal audit services across central government, for example. You can expect to work on cyber security and GDPR projects as well and use competitive tools in the market, helping your clients implement new applications and supporting them in the post-implementation stages. You will be an integral part of the team, involved from the planning to delivery stages of each project.
A successful candidate will have done technology risk or/and IT audit, supporting internal or external audits. It is preferred to have experience within the public sector, although other relevant experience will be considered. You will be CISA qualified or seeking to get qualified with around 3-4 years of experience in IT audit. Our client can also be very flexible for exceptional candidates who have good internal audit experience and are now keen to gear their careers to IT audit.
If you are looking to join a closely-knit team who encourage pursuit of knowledge, personal and professional development including study support then this a great opportunity to consider. Our client proudly offers market-leading mentorship and flexible working which reflects their supportive culture.
Please note: you must be eligible to work in the UK.
The Audit & Risk Recruitment Company – Experts in Audit and Risk Recruitment!


text:
DQ Beauty Supply
Crème of Nature Butter-Licious Curls Hydrating Crème - 7.5oz
Crème of Nature Butter-Licious Curls Hydrating Crème - 7.5oz
- Nurture your natural with this dynamic butter crème.
- Crème of Nature Pudding Perfection Curl Enhancing Creme's lightweight moisturizing butter blend of Argan, Shea and Cocoa butters works to soften, hydrate, and improve manageability for all curl types.
- It reduces dryness and breakage.
- Perfect for daily re-hydration and transitioning styles, this fortifying formula brings life to all curls, plus adds exotic shine.
- Crème of Nature with Argan Oil Butter-Licious Curls does not contain sulfates, parabens, mineral oil and petrolatum.


text:
ICMA 2013 Conference
Keynote Talk
A Robot Co-worker "PaDY"
Dr. Kazuhiro Kosuge
Department of Bioengineering and Robotics Tohoku University, Japan
PaDY (in-time Parts/tools Delivery to You robot) is a
co-worker robot being developed for an automobile assembly process usually
carried out by a human worker. PaDY delivers necessary parts and tools to a
worker when he/she needs them to reduce the worker乫s load, to improve
efficiency of the work and to prevent mistakes of the work. PaDY has been
developed based on the research results of a dance partner robot 乬PBDR.乭 In
this talk, we first introduce why we have started to do the research of the
dance partner robot, which dances a waltz with a human male dancer, where
estimation of the following step lead by the male dancer, or how to read its
partner乫s lead, or how to estimate its partner乫s intention, was one of the key
issues for the development of the robot. We then consider issues for developing
a robot co-worker and introduce PaDY and its prototype system. Experimental
results illustrate how it works and how it could help the worker in the real
Dr. Kazuhiro Kosuge is a Professor in the Department of Bioengineering and Robotics at Tohoku University, Japan. He received the B.S., M.S., and Ph.D. in Control Engineering from the Tokyo Institute of Technology, in 1978, 1980, and 1988 respectively. From 1980 through 1982, he was a Research Staff in the Production Engineering Department, Nippon Denso Co., Ltd. (DENSO Co., Ltd. at present). From 1982 through 1990, he was a Research Associate in the Department of Control Engineering at Tokyo Institute of Technology. From 1990 to 1995, he was an Associate Professor at Nagoya University. From 1995, he has been at Tohoku University. He received the JSME Awards for the best papers from the Japan Society of Mechanical Engineers in 2002 and 2005, the RSJ Award for the best papers from the Robotics Society of Japan in 2005. He is an IEEE Fellow, a JSME Fellow, a SICE Fellow, and a RSJ Fellow. He served as President of IEEE Robotics and Automation Society for 2010-2011 and is currently serving as Junior Past-president of IEEE Robotics and Automation Society.


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The Dr. Pat Show: Talk Radio to Thrive By!: The Ecosystem Approach a revolutionary way to find and ADDRESS what's truly holding you back! with Jason & Patricia Rohn!
07/08/2019 10:00 am PDT
The Ecosystem Approach was created by a two Intuitives who are married to each other! Find out how they got started and learned how to make life better for our clients over the last 20 years! The Ecosystem Approach is based on an amazing discovery that will change the way you think about growth and evolution!
Dr. Pat Baccili
Dr. Pat Baccili helps individuals and organizations break through their limiting Crust so they can reach their unlimited potential. Her specialty is assisting people face t...Find out more »
Jason Rohn
Jason Rohn, Executive Director UTUE, Inc.Jason's breakthrough approach is based on his ability to see that most people use the wrong approach to development and growth...Find out more »
Patricia Rohn
Patricia Rohn, Director UTUE, Inc.As a child Patricia knew she was different. She could perceive things that others could not. Her mother had a similar ability but never fra...Find out more »


text:
Early morning, Helen's friend stops by to say hello.
Ginno's garage. Helen's workplace.
Helen, our brave protagonist.
Mr Bluebeard, the wife-murderer.
Mr Bluebeard's living room exploration.
Mr Bluebeard's tower and his "private room".
Helen discovers the horrible truth.
Final act.
Diorama with the locations of the story.
Reinterpretation of Bluebeard's folktale, now set in 1950s america.
WARNING: contains subject matter that can be triggering to some viewers.


text:
When Ghost Recon Breakpoint launches in October (opens in new tab), it will be available on the Epic Games Store and Uplay, but as the last few Ubisoft titles have done, it will skip Steam. We knew this was coming—it was announced Ubisoft's deal with Epic had been extended back in March (opens in new tab).
PCGamesN confirmed with Ubisoft (opens in new tab) that the PC edition of Breakpoint will only be sold through Uplay and the Epic Games Store. Both The Division 2 and Anno 1800, Ubisoft's last two major releases, have also been sold through Epic's storefront.
The one difference in the case of Breakpoint is that unlike The Division 2 and Anno 1800, Breakpoint won't ever be available to pre-order on Steam.
For Ubisoft's games, presumably including Ghost Recon Breakpoint, the question of which storefront you buy from is slightly academic, since no matter where you purchase the title, you have to play it through Ubisoft's Uplay client.
For more on Ghost Recon Breakpoint, check out Andy Kelly's hands-on impressions here.


text:
By Benjamin Fulford – January 25, 2021
It's over folks, the criminal enterprise known as the Corporation of the United States of America is finished. Actual on the ground reporters with White House Press passes, confirm that nobody is in the White House. The corrupt members of Congress and the Senate dare not show their faces in Washington DC, knowing they could be arrested if they do so.
Read more(excerpt): Hollywood FX Can't Save Bankrupt USA Inc. — Weekly Geo-Political News and Analysis


text: The traditional information on cyber security stands on software and how it's implemented. IoT cyber security provides an extra layer of complexity where the physical and cyber world converges. The wider operational and maintenance situations in the IoT arena require end-to-end device connection that makes the users interact, troubleshoot, login, …Read More »


text:
San Vicente Valley Club's general meeting and luncheon will be on Wednesday, October 26 at the San Vicente Valley Inn, 24157 San Vicente Road.
The luncheon is open to the public; guests are encouraged to make reservations at the front desk of San Vicente Inn or with Mary Jo Adamske (760) 787-1566 or cell (760) 703-0123 by Friday, October 21. The cost is $12.00. If a cancellation is necessary please contact Mary Jo.
Christiane Holmquist is a local, certified landscape designer. She would like to convey the idea that although we have little water to waste, there are ways to create attractive landscapes here. Her presentation, "Fun with Xeriscapes: Water-wise Designs for the Arid Southwest" promises to be entertaining and instructional. She has chosen to present her ideas in October because it is the best time to plant a new garden to enjoy next season
The order of events for the day will be: Social time 11:00, Call to order 11:30, Luncheon 12:00, Program 1:00.
We are a nonprofit organization for all residents of Ramona brought together for the social, philanthropic and civic betterment of the community with our generous charitable contributions toward student scholarships and the needs of our community. For membership information contact Mary Jo Adamske (760) 787-1566 or cell (760) 703-0123.
Other interest groups consist of Book Marks, Bridge Marathon, Lunch Bunch, Friends of the library, Bunco and much more.
Website: http://www.sanvicentevalleyclub.com


text:
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text:
By Ryan Mack
August 27th, 2005 – I was in the bathroom using a washcloth and hand soap to clean the collar of one of my dress shirts. I didn't have the money to use the cleaners so I just used the shirt that was least soiled, and made sure I wore my suit jacket to hide the grease stain on the front right side of the shirt. My dinner was water, which I had freshly boiled the night before and stuck in the refrigerator, along with some doctored up Ramen Noodles. Hot sauce makes anything a fine cuisine when you are hungry. I was preparing for a morning meeting to get my company's first check from Michelle Kulikowski for $300. I didn't have $1.50 for the subway from any other means but the pile of nickels stored in an old shoe in the closet where I had been storing my left over change. The quarters, a rare commodity in these days for me, had been spent many days ago. Yet…I was smiling and wasn't worried.
This was my life as a new entrepreneur. I had seemingly done everything by the book, but the struggles still ensued. Just two years before that day I was making six figures at the largest NASDAQ trading firm in the world. I could afford to eat at the finest restaurants and take trips as I pleased. By the grace of God, I was saving up to purchase a home in NYC so I was amassing the future down payment.
One day…while I was a trader, my aunt called me and asked the question, "What is a stock?" This simple question rocked my entire being. To me, this question meant my own family didn't know what I was doing and really didn't have knowledge of the fundamental principles of the stock market. "How many others beyond my family don't know about stocks?" I thought. This simple question started my quest to pursue a life of educating my family and community of what I was exposed to everyday.
I started to write feverishly every week through a Yahoo chat group I created called "Making Money Work". This wasn't enough to fill my thirst to educate. I turned my Friday Elementary after-work school step team into a financial literacy session about stocks. That wasn't enough. When a young girl was shot and killed by gang members on my block in Brooklyn; I surmised those who pulled the trigger might not have done so if they were educated about other economic opportunities besides a life of crime. It was at this moment I decided to leave Wall Street and pursue a life as a financial planner.
I received a verbal offer to work for a large financial planning firm and turned in my resignation to my Wall Street equities trading firm. On the day I was to finalize the offer, I walked in with my pen ready to sign. I began to brag to the representative, who was a Black man like me, about all of the good I was planning to do as a representative of the firm. "I am going to take this firm to all of those in my community. My church, public housing, gang members, and all over the community," I bragged. He looked me square in the eye and said, "That sounds good…but unless you only work with high net worth individuals everything else is a waste of time."
I was crushed. I felt God speak to me telling me I was in the wrong place. I felt God telling me he had more for me than what this firm had to offer. I was scared because I didn't have any other funds to fall back on accept the funds that I was saving to purchase a home. I left the office saying, "I needed to have my attorneys look over this paperwork!" I didn't have any attorneys, but I did have a message from God there was something greater. If I was going to go into financial planning, I was NOT going to only focus on high net worth and I was NOT going to be a product pusher selling insurance/investments solely for commission. I was going to do what God laid upon my heart to do…educate my people. Knowing that God NEVER provides the vision without provision I rested solely on my faith He would provide.
Now I had to prepare for the blessings to come. I bought four books on how to start a financial planning company. I took a course on how to start a wealth management firm offered by the New York Society of Security Analysts. I asked multiple colleagues who had started similar ventures to mentor me through the process. I quickly realized I had all I needed to be successful and God began to show me the road map to success. I used the funds I was going to use to purchase a home and after I invested $28,000 in funds for the business I had enough left for 12 months of living expenses.
I was ½ month short of my budgeted projections and that brings me back to August 27th, 2005. The next morning, after I dried out my shirt with the newly clean collar by ironing it, I walked with $1.50 in nickels to the subway. I learned the subway only took $1.35 in nickels before it spit out the funds to you. Now I am standing, flat broke, "fronting" with my Burberry Coat on, with the subway token machine looking like an Atlantic City slot machine with all of these nickels spitting back out at me. The crowd behind me sighed in discontent as I held up the line trying to buy a subway pass. I got my money, ran across the street to the store to get a $1 and 2 quarters in exchange for my 30 nickels, put the money in the machine, bought my ticket, took the train to get my check from Michelle Kulikowski, then walked to the nearest bank to cash my first $300 check. I was grinning from ear to ear!
I walked two miles to my friends apartment to borrow $1.50 to get back home, got on the subway, arrived home to boil some more water to drink for dinner, then waited on the check to clear while I was assembling the financial plan of Michelle.
This was my life for the next couple of years. From the "balling" life of Wall Street to the life filled with nothing but faith for something greater (it is when you have nothing but faith you truly realize faith is all you need). I had to use my credit card for the first time as I struggled to make ends meet. I remember going out to eat with my friends and ordering nothing but cranberry juice with extra ice saying, "I just came from a dinner meeting and am not hungry." My stomach growling loudly sounding like a sick cat told them otherwise. Through all of that, I still smiled because I knew that God had something greater for me. He told me I would be speaking to millions of people and I believed Him.
In 2006 I was making enough to make ends meet, but not enough to pay off my debt. So I made the hard choice to let the bill collector wait. I was always able to pay my rent, but there were a few months that I had to make the choice between lights and heat. When my heat was cut off for a week, I put on an extra blanket on the bed and my faith in His word kept me going. When my lights were cut off for a week, I lit some candles and used faith to keep me going. To this day I tell the story that EVERYBODY has a computer because it was during this week I was able to use the computer of the library to continue to work on my business and reach out to new potential clients.
In 2007 I was getting ahead slowly and the clients were becoming much more consistent. Meanwhile, my youth group was steadily growing into a mini-movement. My mentees were gaining a reputation as "those high school teenagers in the community who teach financial literacy to adults". The gang members recognized me and called me "The Money Guy" affectionately and I had helped many of them to form their own businesses that helped to employ other Bloods and a few Crypts and Latin Kings. Working with others in the community we were able to help over 2000 formerly incarcerated men and women create companies and get jobs…I trained all of them for free in financial literacy. Then…on November of 2007 a reporter from Black Enterprise just happened to be in the workshop of my youth. They did a feature story on it and I have always jokingly called myself, "The brokest man to ever be featured in Black Enterprise". It was nothing but God who put that reporter, Diane King, in the audience…she is now a very dear friend.
This was the catapult of many greater things to come. More clients, more exposure, and more hustle using faith as my guide. One day in 2007 I was watching TV and saw Gerri Willis, who at the time was the Personal Finance Contributor for CNN. I told God as I literally yelled at the screen, "I can do what she can do Lord. I am tired of living check to check. I am tired of my feet getting wet because I have walked holes through my good shoes. I want to speak to millions of people like you promised. She is speaking to millions. I am ready!"
That declaration was on a Wednesday. Nine days later on a Friday I ran into Alicia Stewart. Three years before I took her to coffee (although I really couldn't afford that even) and she told me she couldn't afford to pay me for my financial planning services. On this current day she said, "I've been watching the work you have been doing for the past 3 years…it seems you are doing a lot for the community. Have you ever thought about going on CNN?"
"I might have considered it," I said.
"I just got a job as a producer for CNN. Want me to put in a good work for you?"
Twelve days after I was yelling at the TV screen for God to give me something greater, I received a call from the CNN producer. Thirteen days after I was yelling at the screen with Gerri Willis on it, I was ON CNN WITH Gerri Willis talking about financial literacy. Due to the market crash there weren't enough people to talk about the economy so they called me 4-5 days per week, sometimes multiple times per day. The free marketing exposure catapulted me to another level. My phone was ringing off the hook for business and my speaking fees went up to at least $5000 per speech. I paid off my debt and never looked back!
Since…I have written 3 books, co-authored 2, travelled the world teaching financial literacy, still appear on national television teaching principles, speak across the country at various venues about financial literacy, and am doing what God said he would do for me…have me speaking to millions of people teaching His principles of financial stewardship. Nobody can tell me God isn't real and I have many stories I could have shared, but I will wait until my book has a conclusion…because I am just getting started.
Faith is real…faith is tangible. I know it says "unseen", but I am sorry God…I disagree with you on only that thing because I SEE the results of my faith in You everyday! It was faith that allowed me to keep smiling. I wasn't smiling because I was happy to be broke; I was smiling because I knew the brighter day was coming. I wasn't happy WITH my state, my faith allowed me to be happy IN my state. I am learning more and more everyday that faith is more important for others than it is for me, because my faith has allowed me to be a blessing to many others around the world. I regret nothing. I don't regret trading in the millions I could have been making on Wall Street for the days of having to boil water. I continue to have the chance to change lives and exercise my faith for the benefit of others. Thank God I was never worried about the bad news, because God had already delivered the good news. I didn't have to see it in my hands; it's enough for me to have God put in in my dreams to know it is real!
What's next? You will have to wait and see, but just know God has a plan large enough for ALL of us to make the haters very pissed! Believe it and receive it…keep the faith! #Voices4Ed #ChoiceIsOurs


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Over the Thanksgiving weekend, like a main course of ("T"-for-) turkey, I dug into making a start on this build - T(3)-class sub, as-fitted when she sank the IJN cruiser Ashigara, off Sumatra 6/8/45 - using our idea to bash the Revell-Germany 1:144 (German) Type 206A:
Click on Image to Enlarge
At right, this stub was then conservatively - gradually - shaved down (top) until producing a stop at the optimal realignment of the aft hull section - when compared, of course, against the drawing - to which the aft section was then angled and secured, while braced by tape, as shown at middle. At bottom, abaft the (now) keel, the new alignment/attachment was strengthened by beginning to fill the remaining voids - though carefully avoiding any pressure which might force the alignment back open (or crooked).
Even with the above still hardening/setting, already it was possible to begin Step 2: the lengthening of the central hull:
Click on Image to Enlarge
At bottom, mounting collars were fabricated and installed in both openings - aided greatly by first sizing with a piece of paper (white, in background), forming a template for each collar of 0.025" (white) plasticard. Smooth curvature was introduced by rolling each under a smooth drum - e.g., small bottle or spray can - pressed down on a surface having some give - e.g., stacked/folded newspaper, or a mouse pad, etc.. Curvature was made fully complete only during attachment; "spot-welding" with superglue, beginning at the center and working symmetrically outward - 1/8 of the circumference (or less) with each step - while pressing the strip down with a knife edge, flush against the tube. Optionally, after the above - which is not actually "welding", but tacking - the collars can be well and truly - using a "hot" adhesive - welded down, though only along the inside lip, as the outside seam should be kept maximally clean, for subsequent mating with the hull-insert (not shown).
Clearly, you can see that's exactly where this is headed:
(And this is actually not a bad way to depict a cutaway sub, looks like!)
Note the tube looks a bit wider than in the drawing - and this was expected, as the Revell-Germany mold actually scales out 4.2% larger than needed for a 178-scale T(3). (Additionally, the macro-mode of my camera, above, may further accentuate the effect.) Not to worry in any case, however: once stoutly constructed, in the proper alignment, the entire hull can if need be pinched back 4.2% narrower. Only if needed, that is - since it's entirely possible and even likely that the overlying saddle tanks, top casing and heavy keel could, between them, hide this discrepancy so much as to basically render it (visually) undetectable.
Meanwhile, the required hull insert - another simple tube, its diameter determined by fitting against the existing tube-ends, and length determined by overlay directly on the drawing, as shown - can now be fabricated and installed. However at this point I had to break off work.
Look for exactly that last - and more work - Coming Soon. Happy Thanksgiving, y'all!
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Indictments: Former officers coerced sex; ignored cases
COLUMBIA, S.C. - Four former South Carolina police officers who worked in the criminal investigations division for Horry County coerced women into sex or ignored cases they were supposed to be investigating, according to indictments released Friday by a state grand jury.
At least four lawsuits have been filed against one of the officers, Troy Large. One of the lawsuits accuses him of ignoring the case of a woman who said she was raped and then demanding she participate in a nude catfight or else he would make sure she lost visitation with her children.
Large is charged with six counts of misconduct in office and five counts of third-degree criminal sexual conduct. The South Carolina Attorney General's Office didn't immediately release the indictments against him, but said in a news release that Large coerced women into sexual acts and had inappropriate relationships with victims in cases he was investigating.
Large denied coercing women into sex in a deposition in one of the lawsuits. He couldn't be reached for comment Friday.
The three other officers indicted Friday all face misconduct in office charges.
- Darryl Williams didn't fully investigate crimes he was supposed to, the indictment said. He is charged with nine counts of misconduct in office.
- Todd Cox closed cases when the investigations weren't finished, prosecutors said. He is charged with 16 counts of misconduct in office.
- Luke Green had a sexual relationship with a police informant and had sexual contact with a suspect he was arresting, according to indictments. He is charged with three counts of misconduct in office.
It wasn't known if any of the former officers had lawyers.
The State Law Enforcement Division began investigating Horry County Police after county leaders discovered the allegations, Horry County Administrator Chris Eldridge said in a statement.
The county made significant changes in the police department's criminal investigation division, which now has a new captain and deputy chief of operations. Horry County Police Chief Saundra Rhodes also abruptly retired.
"Horry County Police Department starts with new leadership at the top this Monday with the arrival of Chief Joe Hill," Eldridge said. "Chief Hill's focus will be continuing the review of accountability measures for officers."


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Mandy Moore To Sing On 'Extreme Makeover: Home Edition'
Mandy Moore will appear on the season finale of ABC's 'Extreme Makeover: Home Edition' on May 20th, where the singer/actress will sing to camp kids at the Camp Heartland in Putnam County, New York. The family involved features a single mother raising three kids, two of which are HIV positive.
DJ AM 'Was Really In Love With' Mandy Moore
March 27, 2007 – Though DJ AM has been seen hanging out with model Jessica Stam, The New York Post hears the club magnet is taking his breakup with Mandy hard. "AM was really in love with Mandy," a spy explained, "but they just had such different lifestyles, and they couldn't make it work."
Mandy Moore Visits Minneapolis' KDWB, Reaches Out To Britney
March 25, 2007 – Mandy was in studio with Dave, Corey and Steve-O of KDWB in Minneapolis on Tuesday (March 20). Moore talked about her movie projects, working with Robin Williams, her thoughts on Britney Spears and how "I would love to be a friend" to support the singer, dealing with paparazzi in Los Angeles, and making sure she's not surrounded by "yes" people. The interview audio at akamai.net has since been removed.
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i HOPE MANDY SEE ALL ME MSG


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Many misconceptions, apprehensions and perceived barriers about China continue to abound in the West, despite China's now well established position as the number two economy in the world. These are further fuelled by a lack of basic appreciation and understanding of Chinese culture, society, ways of thinking and ways of doing. Education stands to play a unique role in bridging some of these gaps, presenting opportunities to those that are up for the challenge of going to China for study on exchange, language learning, or full degree programs. For those coming from western countries, studying in China offers an experience that will challenge individual understandings and sensibilities in ways more conventional study abroad destinations (US, UK, Canada, Australia, New Zealand) never could.
Traditional criteria often considered in the process of selecting the right MBA program and institution include networks, institutional reputation and prestige, and of course, the courses taught. These criteria apply as much in China as anywhere else. There are other reasons however why embarking on this journey in China is worthy of consideration. In a global economic environment where China's role and prominence is on the rise, there is a great demand for business professionals who are able to comfortably operate in both western and Chinese environments. This relates not only to foreign companies wishing to open or expand operations in China. It is relevant also as China extends its reach into foreign markets to explore opportunities for growth.
MBA programs in China are offered in English or Mandarin and typically take 1.5 – 2 years to complete. Leading programs include CEIBS (http://www.ceibs.edu) and Tsinghua University's MBA (http://mba.sem.tsinghua.edu.cn/mbaen). Another model that is not uncommon is the BiMBA program offered by Peking University (http://en.bimba.edu.cn). Here one of China's most prestigious institutions has partnered with one of Europe's best managements schools (Vlerick Leuven Gent Management School) to deliver a program in Beijing by western and Chinese professors.
While international student numbers are on the rise, as an international student embarking on your MBA in China you can generally still expect to be in the minority. And this is where the real learning begins. MBA programs involve large amounts of teamwork. Fundamental cultural differences that extend way beyond language barriers mean it will be no time before you will naturally be suggesting going to lunch before talking shop and the assignment at hand. You will learn to listen before you speak and approach problems from different perspectives, as your team mates invariably will not share the same views as you. You will realise eventually that there is no silver bullet, and that the same challenges as you faced them on the first day still exist, with many more uncovered along the way. Only now you have an appreciation and understanding of the environment and a set of tools to navigate differences in thought and practice, culturally or otherwise, that no number of journal articles could ever hope to provide you with.
Put simply: if you want the easy, comfortable route to obtaining your MBA, and if learning macro economic theory from those who developed it is a priority to you, enrol with one of the Ivy League institutions where you will network with the up-and-coming western elite and learn from the West's leading minds on the subjects at hand. If however you want to be challenged more broadly, grow and develop in ways you didn't know existed, and don't mind sacrificing some of the more traditionally held notions of what an MBA should be about, then consider doing your MBA in China.


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Riad Le Bel Oranger
When in Morocco, there is no better place to stay than in a Riad or a Dar. Both are traditional Moroccan house mainly found in Marrakech, Fes, Tangier, Casablanca, etc. These houses are build based on Islam architecture that ensures privacy of the residents. You might notice that many old houses in Morocco does not… Read More Riad Le Bel Oranger


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Falls Church, VA—October 27, 2016—The Farm-to-Consumer Legal Defense Fund Foundation (FTCLDFF), a 501(c)(3) sister organization to the Farm-to-Consumer Legal Defense Fund (FTCLDF), has begun operations. The main purpose of FTCLDFF is to support organizations such as FTCLDF in the work to protect and expand farm and food freedom. FTCLDFF received 501(c)(3) status (EIN 47-3917847) from the Internal Revenue Service (IRS) earlier this year. Donations to FTCLDFF are tax-deductible to the fullest extent allowed by law. Donations to FTCLDF are not tax-deductible.
FTCLDFF will provide financial support for litigation and other legal work done by FTCLDF attorneys. FTCLDFF will also be developing and providing educational resources for its supporters and the general public. In addition, FTCLDFF will be providing compassionate relief to small farmers and local food producers suffering hardship.
As a reminder of the fight we are in over freedom of food choice, here are statements the United States Food and Drug Administration (FDA) made in a reply brief responding to a lawsuit filed by FTCLDF challenging the interstate raw milk ban:
- "There is no absolute right to consume or feed children any particular food"
- "Plaintiff's assertion of a 'fundamental right to their own bodily and physical health, which includes what foods they do and do not choose to consume for themselves and their families' is similarly unavailing because plaintiffs do not have a fundamental right to obtain any food they wish."
When the founding fathers drafted the Constitution, there was nothing in the document about freedom of food choice because it was a right that was taken for granted. With the harassment farmers, artisan food producers, and homesteaders have undergone from federal, state, and local government agencies, that has long stopped being the case. FTCLDFF was formed to change that through its support of efforts to establish the right of all people to obtain the food of their choice from the source of their choice.
This is the right fight. Those interested in donating can go here.
Media Contact:
Pete Kennedy, Esq.
[email protected]
(703) 208-3276
Services provided by FTCLDF go beyond legal representation for members in court cases.
Educational and policy work also provide an avenue for FTCLDF to build grassroots activism to create the most favorable regulatory climate possible. In addition to advising on bill language, FTCLDF supports favorable legislation via action alerts, social media outreach, and the online petition service.
You can help FTCLDF by becoming a member or donating today.
8116 Arlington Blvd, # 263
Falls Church, VA 22042
Prefer to make a tax-deductible donation? Donate online at bit.ly/NFG4FTC.
You may also contact us by email at [email protected] or call 703-208-FARM (3276). Thanks for your support.


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The Bobby Massie Authentic Jersey Hardaway deal was an unnecessary overpay. Season Ticket Holders!The team made the news official on Saturday.The 26-year-old continues to put together a breakout campaign for Oakland.One of the main things Washington's prospects did off the ice was going to many different restaurants around D.C.
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The universe of ELC players proves this example wasn't anecdotal.The NHL sort of apologized for this on Friday.According to the research, eldest children are most likely to become scientists or engineers.It's also vital to make sure your butter is kept in the fridge �?or at least, well-covered �?during the cooking process, to avoid a pesky case of accidental salmonella.
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Despite being out since Dec.U16 Euro Championship B.Matthews needs to be on the ice for the OT faceoff because, first of all, it's worked in the past.They lead the dreaded Metropolitan Division by just a single point over the New Jersey Devils going into Saturday's games.The team came back from 2 down but needed a 93rd-minute equalizer from Matt Doherty to draw 2, forcing a replay.
Only Jason Hatcher of the Dallas Cowboys finished with more sacks than Casey among defensive tackles in 2013.Mattheos captained the Wheat Kings in 2018 and led the team in goals, assists , points, shots on goal , power-play goals and power-play assists .Tebow had developed a relationship with Shula, but Meyer was about make a big impression on Tebow.High-end small cruise ships charter the waters, taking passengers to see the striking rock formation at the river's mouth and its waterfalls, as they spot the stealth-like salties.Dudley was ejected and Shaq was hit with a technical foul.
There is also the possibility that there could be intra-plane gaming tournaments.Since then, he's become as irreplaceable as Mesut Ozil.Treinen has now been touched up for runs in each of the last two outings after serving up a pair of home runs to Ian Happ and Javier Baez on http://www.shopofficialpacersonline.com/Authentic-Myles-Turner-Jersey Monday.Rebecca Long-Bailey, Labour's spokesperson on business, told the BBC at the weekend that any deal with the Conservatives must be entrenched so that a future Conservative leader wouldn't be able to rip up the changes that have been agreed: in other words, Boris-proofed.
He is a special athlete.I see the Cowboys in the middle of this.I believe in this team, Kanter said.That puts a strain on the cap space, just as you are hoping to add complementary pieces around your young players.Others have social-media platforms to share their daily life.
I was just so caught up in the moment and the experience.Foles needs to find the right system to play in, but isn't that true of any quarterback?U16 Euro Championship A.The population reached close http://www.officialauthenticbearstores.com/Nike-Bobby-Massie-Jersey.html to zero in 1920.
Daytona's second race, which traditionally hovered around the Fourth of July, will now be pushed back to Saturday, Aug.


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I'm thinking long-term and with GM Stories around 1-3 Steps, I worry that continuing to give out 1 and 2-pt Rewards may result in all Heros becoming homogenized in that everyone will have all Skills at Rank 1 and all 1-pt Advantages, even if they don't 'want' them. I've had some players tell me that there are skills they don't want to have any Ranks in and not everyone needs/wants to be a Linguist or Able Drinker.
That said, I was wondering if anyone has thought of alternate GM Story Rewards that don't involved XP?
Some examples I've used in the past for other games, including 7th Sea 1st...(A lot of these are 'limited' Advantages or similar)
1) [Good / Bad] Reputation with [Town / Individual / Group]: Gain (or Lose) 1 Dice from all Social Risks when dealing with the affected NPCs
2) Local Heroes with [Town / Individual / Group]: When purchasing items from the affected NPCs, you are treated as having +1 Wealth to spend each Game Session
3) Local Reputation with [Town / Individual / Group]: You are considered to have the reputation [Reputation] when dealing with the affected NPCs
4) Informant [Name]: This person will often have information that is useful if you can locate them and pay them appropriately
5) Confidante [Name]: This person can accomplish a single task for you if you are in the same location but will not put themselves at risk
6) Ally [Name]: This person can accomplish a single task for you if you are in the same location, including putting themselves in harm's way. They may also come to you if requested.
Any thoughts or have others done this?


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Romans 6:1-14
Can we really walk in victory against sin in our lives? Romans 6 gives the liberating truths needed to begin the walk of victory. Many Christians are simply unaware that they are free!
Many Christians continue enslaved to sin– ignorant of the freedom available to them through Christ's death and resurrection.
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