text:
Many monoclonal antibodies (mAbs) and other protein drugs have targets usually residing within tissues, making tissue concentrations of mAbs relevant to their pharmacologic effects. Therefore, knowledge of tissue distribution kinetics is important to better understand their pharmacokinetics and pharmacodynamics. The tissue distribution of mAbs is affected by many physiologic factors that may be altered in disease status. In the present work, we studied the tissue distribution kinetics of the fusion protein etanercept in inflamed joint tissues and examined the impact of inflammation on the tissue distribution of etanercept. Etanercept concentration profiles in plasma, blister fluid, and different tissues were obtained from healthy and collagen-induced arthritic (CIA) rats by use of a fluorescence quantification method via IRDye800CW labeling. Stepwise minimal and full physiologically based pharmacokinetic (PBPK) approaches were applied to characterize the distribution kinetics of etanercept in tissues in healthy and diseased animals. Etanercept exhibited modest tissue access (tissue/plasma area under the concentration curve [AUC] ratios 0.03–0.15 and estimated tissue reflection coefficients [σ] of 0.6–1.0), but with good penetration into arthritic paws (tissue/plasma AUC ratio 0.23 and σ 0.36). Etanercept exposure in the inflamed paws of CIA rats was approximately 3-fold higher than in normal paws taken from either CIA or healthy rats (tissue/plasma AUC ratios 0.23 versus 0.07 and σ 0.36 versus 0.71). The tissue distribution kinetics of etanercept in arthritic paws were well characterized with PBPK modeling approaches. Etanercept shows good penetration to arthritic paws in CIA rats. Our study indicates that inflammation produced increased tissue distribution of etanercept in CIA rats.
Rheumatoid arthritis (RA) is a chronic inflammatory disease exhibiting persistent inflammation, hyperplasia, and infiltration of immune cells in the joint synovial tissues (Choy and Panayi, 2001). The soluble cytokine tumor necrosis factor α (TNF-α) plays a pivotal role in the pathogenesis of RA. TNF-α is secreted by mononuclear cells in synovial fluid and interacts with TNF receptor locally. Through autocrine and paracrine mechanisms it exerts a proinflammatory effect (Brennan et al., 1992). In patients with active RA, serum and synovial fluid TNF-α concentrations are elevated compared with healthy subjects (Hopkins and Meager, 1988; Edrees et al., 2005). Etanercept is used for the treatment of RA by specifically binding to TNF-α with high affinity, neutralizing it and therefore blocking its proinflammatory activity in synovial fluid (Culy and Keating, 2002).
Etanercept is a dimeric fusion protein consisting of two ligand-binding domains of human p75 tumor necrosis factor receptor and the human IgG1 Fc portion (Scott, 2014). The plasma pharmacokinetics of etanercept have been well studied in various species (Lon et al., 2011; Zhou et al., 2011). However, information on the distribution kinetics of etanercept to inflamed joint tissues is lacking. As the drug concentration at the target site determines the magnitude of the pharmacologic effects, knowledge of etanercept distribution in the joints is important to better understand the pharmacokinetics and pharmacodynamics. With a molecular size of 150 kD and high hydrophilicity, etanercept is likely to show limited tissue distribution, making its concentration in inflamed joint tissue a major determinant of the pharmacodynamic effects.
The tissue distribution of monoclonal antibodies (mAbs) and other protein drugs is affected by physiologic factors such as blood flow, tissue permeability, and the expression and turnover rate of their target antigens (Tabrizi et al., 2010). Therefore, changes in physiologic status may alter the tissue distribution of these macromolecular drugs. RA causes increased vascular permeability as well as joint tissue edema, which would lead to increased tissue distribution of large molecules (Bell et al., 1983). Although the plasma pharmacokinetics of etanercept is not significantly altered in patients with RA compared with healthy subjects (Zhou et al., 2011), whether RA will alter the tissue distribution of etanercept has not been investigated and remains unclear.
We investigated the tissue distribution of etanercept to the inflamed joint tissue and examined the impact of inflammation on the kinetics of this distribution using the collagen-induced arthritic (CIA) rat model. This is a well-established animal model that mirrors human RA. We previously studied the plasma pharmacokinetics of etanercept using this animal model (Lon et al., 2011), and the present study extends our work by examining the tissue distribution kinetics of etanercept.
Materials and Methods
Etanercept (Immunex Corporation, Thousand Oaks, CA) was diluted with vehicle (10 mg/ml sucrose, 5.8 mg/ml sodium chloride, 5.3 mg/ml l-arginine hydrochloride, 2.6 mg/ml sodium phosphate monobasic monohydrate, and 0.9 mg/ml sodium phosphate dibasic anhydrous, pH 6.3 ± 0.2) and was stored at 2–8°C before use.
IRDye800CW Labeling.
Etanercept in the injection solution was first subjected to a buffer exchange with phosphate-buffered saline (PBS), pH 7.8, using Zeba desalting spin columns (5 ml; Pierce Biotechnology, Rockford, IL) followed by dilution to 1 mg/ml. IRDye 800CW Protein Labeling Kit–High Molecular was used for etanercept labeling according to the manufacturer's instructions (LI-COR Biosciences, Lincoln, NB). Briefly, 200 µl of 1 M potassium phosphate buffer (pH 9.0) was added into 2 ml of 1 mg/ml etanercept in PBS to adjust the pH to 8.5. Dye was resuspended in water (25 μl) and 15.5 μl was added dropwise to the etanercept solution, then it was incubated at room temperature for 2 hours in the dark. Free dye was then removed by Zeba desalting spin columns. The IRDye800CW-etanercept (I-E) conjugates were stored at −20°C until use.
Characterization and Stability of IRDye800CW-Etanercept (I-E) Conjugates.
To calculate the dye/etanercept (D/P) ratio and concentration of I-E conjugates, the conjugates were diluted with PBS/methanol (9:1) to determine the UV absorbances (A) at 280 and 780 nm. The following equations were applied in these calculations.Dye/etanercept ratio:(1)Final etanercept concentration:(2)where εdye is the molar extinction coefficient of the IRDye800CW at 780 nm, which was 270,000 M−1 • cm−1 according to manufacturer's product manual. The molar extinction coefficient of etanercept (εetanercept) at 280 nm is assumed to be equal to that of IgG1 (203,000 M−1 • cm−1).
Ten percent denaturing SDS-PAGE was performed to examine the conjugation of etanercept with IRDye800CW. The stability of IRDye800CW-labeled etanercept in plasma was examined by incubation in normal rat plasma. Briefly, the stock solution of labeled etanercept was spiked into plasma and incubated at 37°C for up to 15 days. The spiked plasma was sampled over time, and the fluorescence was measured in a 96-well plate using the Odyssey Infrared Imaging System (LI-COR Biosciences).
Male Lewis rats, aged 6 to 9 weeks, were purchased from Harlan (Indianapolis, IN). The animals were housed individually in the University Laboratory Animal Facility and acclimatized for 1 week under constant temperature (22°C), humidity (72%), and 12-hour light/dark cycles. Rats had free access to food and water. All protocols followed the Principles of Laboratory Animal Care (NRC, 1996) and were approved by the University at Buffalo Institutional Animal Care and Use Committee.
Induction of CIA in Lewis Rats.
The induction of CIA in Lewis rats was performed following procedures described previously elsewhere (Earp et al., 2008; Lon et al., 2011). The required reagents were purchased from Chondrex (Redmond, WA). Thirty Lewis rats were used for induction, and around 60% successfully developed CIA in one or both hind paws.
Suction Blister Technique.
A suction technique was applied to obtain skin blister fluid. Previous studies have shown that under well-controlled experimental conditions, the composition of sampled blister fluid closely resembles interstitial fluid (ISF) (Kiistala, 1968; Herfst and van Rees, 1978). Therefore, blister fluid was assumed to serve as a surrogate for skin ISF. An acrylic device for producing suction blisters was manufactured in-house that was similar to the device used for sampling human blister fluid reported by Kiistala (1968). Modifications were made to the plate to fit the skin area on the back of the rats. A Welch Gemini dry vacuum pump model 2060 (Welch-Ilmvac, Niles, IL) was used to provide constant pressure. A picture of the device and the dimensions of the plate are shown in Fig. 1. The widths of the edges as well as the distance between the holes of the plate were carefully designed to minimize possible skin damage.
The animals were subjected to epilation using Nair hair removal lotion (Church & Dwight Co., Ewing, NJ) 1 day before suctioning. Smooth skin was observed with no rash.
To perform blister suctioning, the animals were anesthetized by inhalation of 2% isoflurane, and the plate was placed on the smooth skin area on the upper back of the epilated rat. Medical tape was used to wrap the plate to make sure it sealed well on the skin surface. The pump was turned on, and the pressure was gradually tuned to −150 mm Hg and maintained for 2 hours. Upon completion of suctioning, the plate was gently removed, and the blister fluid samples were collected with a 29 ga/1/2 ml syringe.
Pharmacokinetics of IRDye800CW-Etanercept Conjugates.
CIA rats were used 21 days after induction, which corresponds to peak edema in arthritic paws. Healthy (n = 18) and CIA Lewis rats (n = 17) weighing around 250 g were dosed with 5 mg/kg I-E conjugates subcutaneously (s.c.). The suction blister technique was applied at 6 hours, 12 hours, and 1, 2, 4, 7, and 11 days. At each time point, three animals were sampled for blister fluid (except for day 7, when two CIA rats were sampled). After blister fluid was collected, the rats were sacrificed by exsanguination from the abdominal aorta, and blood samples as well as tissue samples (liver, heart, spleen, lung, kidney, adipose tissues, gastrocnemius muscle, and paw) were harvested. The blister fluid samples were kept on ice and stored at −80°C. The blood samples were immediately centrifuged at 2000g, 4°C, for 15 minutes. The plasma fraction was then obtained, aliquoted, and stored at −80°C. The tissue samples were immediately placed in liquid nitrogen and stored at −80°C.
Tissue Protein Extraction.
Frozen tissue samples were pulverized in liquid nitrogen. Preweighed tissue powder was placed in tissue protein extraction solution (DSTPES) purchased from Dualsystem Biotech (Schlieren, Switzerland) containing 0.5% complete mini-protease-inhibitor (Roche Applied Science, Vilvoorde, Belgium) (1 ml/175 mg tissue). The resuspended tissue was shaken for 30 minutes at room temperature. Supernatants were obtained after centrifugation at 20,000g, 4°C, for 15 minutes and stored at −80°C.
Quantification of IRDye800CW-Etanercept Conjugates by Fluorescence.
Briefly, the stock solution of I-E conjugates was spiked into diluted blank plasma or blank tissue protein extracts to prepare a set of seven standards (ranging from 7.8 to 500 ng/ml). Plasma samples were diluted 50-fold with PBS, and blister fluid samples were diluted 10- or 50-fold with PBS to the desired linear concentration range. The tissue protein extracts were directly measured without dilution except for those from the kidney (1/4 dilution in PBS). Fluorescence (excitation = 778 nm; emission = 800 nm) of each 75-μl sample and standard was measured in a 96-well plate with the Odyssey Infrared Imaging System. The concentrations in unknown samples were determined from the standard curve. Due to the loss of intensity of fluorescence over time, the measured concentrations in plasma, blister fluid, and tissues were corrected with a biexponential decay function based on the in vitro stability of I-E conjugates in plasma:(3)
The plasma concentration of the labeled etanercept was also quantified by enzyme-linked immunosorbent assay (ELISA) for validation purposes. The ELISA procedures were described previously elsewhere (Lon et al., 2011).
Tissue protein extracts were further subjected to 10% SDS-PAGE to determine the fluorescence intensity of the intact conjugate band as well as the fragment bands. The estimated fraction of the intact conjugate was used to calculate intact labeled etanercept concentrations in tissues.
Data Analysis.
The area under the concentration time curve (AUCinf) of I-E conjugates in plasma, blister fluid, and tissues was estimated by the trapezoidal rule using WinNonlin 6.1 (Phoenix, Pharsight Corporation, Palo Alto, CA).
Mathematical Modeling.
A stepwise modeling approach was applied to characterize the plasma and tissue disposition of etanercept. First, plasma concentration profiles from the present study together with our previously published data (Lon et al., 2011) were fitted with a minimal physiologically based pharmacokinetics (mPBPK) model to describe the disposition of etanercept. The model includes plasma, lymph, and two lumped tissue compartments connected in an anatomic manner, as shown in Fig. 2 (Cao et al., 2013). The nonlinear absorption and mixed linear and Michaelis-Menten elimination of etanercept in CIA rats were assumed based on previous studies (Lon et al., 2011). The linear clearance was assumed for healthy animals for simplicity. The model is described by a set of differential equations:(4)(5)(6)(7)where Cp is the plasma concentration in Vp (plasma volume) and Ctight and Cleaky are ISF concentrations in two types of lumped tissues categorized by the leakiness of vasculature (continuous and fenestrated). The Vtight (0.65 • ISF • Kp, where Kp is the available fraction of ISF for antibody distribution) and Vleaky (0.35 • ISF • Kp) are ISF volumes of the two lumped tissues, and Vlymph is the lymph volume, which is assumed equal to blood volume. L is the total lymph flow rate, and L1 and L2 account for 1/3 and 2/3 of the total lymph flow. The σ1 and σ2 are vascular reflection coefficients for leaky and tight tissues. The σL is the lymphatic capillary reflection coefficient assumed to be 0.2.
For CIA rats:(8a)For healthy controls:(8b)
Etanercept is assumed to be eliminated from plasma, and CLdis and CLhea represent the linear clearances in CIA and healthy animals. An additional Vmax/ Km term was used to account for the nonlinear clearance in CIA rats.
The input functions are:(9)where(10a)(10b)
The absorption of etanercept is described as zero-order (k0) followed by first-order absorption (k1) after the time period τ, which is consistent with previous studies (Lon et al., 2011). The F is the bioavailability for s.c. administration. The Fr represents the fraction of the dose that undergoes first-order absorption, and ka is the absorption rate constant.
In the next step, parameter values estimated from the previous model were fixed to fit the blister fluid concentration profiles in healthy and CIA animals. The blister compartment was incorporated into the mPBPK model as shown in Fig. 3. The concentration of etanercept in blister fluid can be described as:(11)where Cbf represents the etanercept concentration in blister fluid, Vbf is the total volume of blister fluid yield during the duration of suction (tdur = 2 hours), σski is the vascular reflection coefficient of the skin. The inflow of the blister fluid compartment is derived by dividing Vbf by tdur, and the outflow is negligible.
Finally, a modified full PBPK model was developed to characterize the distribution of etanercept in various tissues, including healthy and arthritic paws. Major organs and tissues included in this model are liver, lung, kidney, spleen, heart, skin, muscle, adipose, carcass remaining tissues, and paw. Figure 4 shows the model structure. The modified PBPK model is based on previous published PBPK models but with some simplifications (Garg, 2007). Tissue ISF is assumed to be the major extravascular space for mAb distribution. The distribution of etanercept from plasma to tissue interstitial space is dominated by convection. Lymphatic drainage is the primary pathway for tissue clearance. Shown are the mass balance equations for plasma, lymph, and tissue compartments:(12)(13)(14)where Ctis_isf is the tissue ISF concentration, the Visf and Ltis are the ISF volume and lymph flow rate of each tissue and organ, and σtis is the vascular reflection coefficient of a specific organ.
The measured tissue concentration is total tissue concentration; therefore, the observed tissue concentration is:(15)where Ctis is the total tissue concentration, Vres is the volume of residual plasma in the tissue, and Vtot is the total tissue volume.
A normal paw weighs around 0.5 g in the rat with a body weight 280 g. The arthritic paw size is about 1.5-fold greater due to inflammation and edema. In the model, the tissue volume and the available volume of ISF for etanercept distribution in arthritic paw increases by 0.5-fold of the ISF volume in normal paws (Kp • ISFnormal + 0.5• ISFnormal).
The physiologic parameters for plasma, lymph, and tissue ISF and total volume were obtained from the literature (Shah and Betts, 2012). The residual plasma volume was assumed to be 6% of the total tissue volume for lung and spleen, 3% for kidney, liver, and heart, and 1.5% for muscle, paw, and adipose. This assumption is based on the average measurements from the literature and assuming the hematocrit is 45% in rats (Bernareggi and Rowland, 1991; Baxter et al., 1994; Garg, 2007). The total lymph flow rate was allometrically scaled from human (2.9 l/day) with a factor of 0.74. The lymph flow rates of tissues were derived from the total flow rate based on the fraction of their blood flow rates. The ISF volume of paw was obtained by assuming that 1 g of tissue yields 1 ml volume and that ISF accounts for 15% of total paw tissue volume. The physiologic parameter values used for model fitting are listed in Table 1.
Model fittings were performed by nonlinear regression analysis using the maximum likelihood algorithm in ADAPT 5 (D'Argenio et al., 2009). The variance model used is:(16)where Vi is the variance of the ith observation, σa and σp are additive and proportional variance model parameters, and Yi is the ith model prediction. The goodness-of-fit was evaluated by objective function values and diagnostic plots.
IRDye800CW Labeling and Stability of IRDye800CW Etanercept Conjugates.
Etanercept conjugated with IRDye800CW exhibited a D/P ratio of 2.5:1. The measured concentration of the I-E conjugates was 1.01 mg/ml, which is close to the etanercept concentration (1 mg/ml) used in the labeling reaction. This suggests that there was no loss of etanercept during the reaction and purification process.
The I-E conjugates were subjected to SDS-PAGE to characterize the conjugation of IRDye800CW with etanercept. As shown in Fig. 5A, a clear single band of IRDye800CW etanercept conjugates was detected, which corresponds to a single arm of etanercept (75 kDa). This indicated that etanercept was labeled with IRDye800CW successfully and no degradation or free dye occurred.
In the stability study, IRDye800CW labeled etanercept was incubated in plasma at 37°C for over 15 days. The measured fluorescence showed a loss of intensity over time (Fig. 6), which could be characterized by a biexponential function (eq. 3). SDS-PAGE was performed with the samples, and the results showed no sign of degradation (data not presented). This indicated that the loss of fluorescence intensity is not likely caused by degradation of the conjugates.
Quantification of IRDye800CW Etanercept Conjugates.
The I-E conjugates in plasma and tissues were measured by fluorescence and adjusted using the biexponential decay function. Plasma concentrations of etanercept were also determined by ELISA for comparison and validation. Measurements by fluorescence suggested altered plasma pharmacokinetics (less etanercept exposure). However, after adjustment using the decay function, the plasma concentration profile was consistent with that measured by ELISA and with our previous pharmacokinetic studies of etanercept (Lon et al., 2011). The measured tissue concentrations of I-E conjugates by fluorescence indicated significantly increased liver and kidney uptake, which was partially attributed to the presence of a greater amount of IRDye800CW protein fragments, as shown in the SDS-PAGE results in Fig. 5B. The tissue concentrations of the intact conjugate were then estimated with the relative percentage of fluorescence intensity of the intact conjugate band.
Pharmacokinetics of IRDye800CW-Etanercept Conjugates.
The AUC of intact I-E conjugate concentrations in all tissues as well as plasma and blister fluid concentrations for healthy and CIA rats were first determined. The parameters are listed in Table 1. Slightly increased clearance of I-E conjugates was observed in CIA rats. Exposure of I-E conjugates in blister fluid was 1.5-fold greater in CIA rats compared with healthy controls. In both healthy and CIA rats, the biodistribution of I-E conjugates was more extensive in liver, spleen, lung, kidney, and heart, with tissue/plasma AUC ratios of 7%–15% compared with fat and muscle (tissue/plasma AUC ratios of 3%–6%). Etanercept exposure in inflamed paws in CIA rats was 3-fold higher than in normal paws (tissue/plasma AUC ratios of 0.23 versus 0.07).
The stepwise modeling approach was used to characterize tissue biodistribution of I-E conjugates in healthy and CIA rats. In the first step, plasma concentration profiles in healthy and CIA rats were fitted with the mPBPK model to describe the disposition characteristics of I-E conjugates. Model fittings of the plasma concentration profiles are presented in Fig. 7, and parameter estimates are listed in Table 2. The estimated values of the parameters related to s.c. absorption correspond well with previous results (Lon et al., 2011). The mixed clearance mechanism of etanercept in CIA rats was assumed in the present model, which gave more flexibility in describing the elimination of etanercept. The estimated vascular reflection coefficients (σ1 and σ2) suggest a moderate interstitial distribution of etanercept.
In the next step, an additional blister fluid compartment was included in the mPBPK model to characterize labeled etanercept concentration profiles in blister fluid in CIA and healthy rats. Parameter estimates from the previous step were fixed. The model-fitted profiles are presented in Fig. 8, and parameter estimates are listed in Table 2. Despite the variability of the measurements, the model is able to describe the general trend of etanercept concentration profiles in blister fluid. The σski in healthy animals is slightly greater than that in CIA rats, indicating a slight increase in vascular permeability in CIA rats compared with healthy controls.
The full PBPK model was finally applied to describe the tissue distribution of I-E conjugates in healthy and CIA rats. Total tissue concentrations were used for model fitting, which is represented in the model output as the drug amount in the residual plasma and interstitial spaces divided by the total tissue volume. Residual plasma volumes were assigned using values within the literature-reported range (Table 3). Model-fitted profiles are presented in Fig. 9 for CIA rats and Fig. 10 for healthy controls. Parameter estimates are listed in Table 3.
There were no significant differences in tissue exposures in liver, lung, heart, kidney, and spleen in healthy and diseased animals. The AUC values showed slightly increased tissue distribution in CIA rats in muscle and adipose tissue. However, the model was not able to differentiate the change in permeability in these tissues in CIA rats. In general, etanercept shows modest tissue distribution consistent with mAbs. In tissues such as lung, liver, kidney, spleen, and heart, the distribution of etanercept was more extensive (σ values are in the range of 0.7–0.9) compared with muscle and adipose (σ values are greater than 0.9). The kidney σkid value could not be estimated and was fixed to 0.8 due to the poor quality of the kidney data.
With inflammation, arthritic paws are subject to increased extravascular protein infiltration and edema. Increased extravascular protein infiltration is caused by increased vascular permeability. This is reflected as a change in vascular reflection coefficient σ in the model. Paw edema is due to the buildup and retention of fluid in the extravascular space caused by inflammatory responses. Arthritic paws exhibited edema and showed on average a 1.5-fold increase in volume as indicated by measured paw sizes. The model assumed a 1.5-fold increase in paw ISF in arthritic paws with the increase reflecting the free fluid phase (the volume of the gel phase does not expand in edema) (Wiederhielm, 1979). Therefore, the interstitial volume for etanercept distribution is (Kp • ISFnormal + 0.5 • ISFnormal). Also, the lymph flow rates, considered as the clearance of ISF, are correlated with interstitial volumes at equilibrium. Therefore, in arthritic paws the lymph flow rate is also increased by 0.5-fold. Figure 11 shows the fitted concentration curves in arthritic and normal paws.
Similar etanercept distribution profiles have been observed in paws from healthy controls and nonaffected paws from CIA rats. Arthritic paws showed increased etanercept exposure compared with normal paws. The estimated values of σpaw_dis and σpaw_hea indicate a 2-fold increase in vascular permeability in arthritic paws. This corresponds to a simulation study conducted previously, which suggested that a 4-fold change of vascular permeability will cause a 3-fold increase in ISF volume (Wiederhielm, 1979). The lymph flow rate in normal paws is estimated to be 0.01425 ml/h, which is 10-fold faster compared with the same mass amount of muscle. This may be due to either insufficient data at the early time points or the frequency of muscle movement in the paw.
Overall, etanercept exhibits modest tissue distribution as suggested using both mPBPK and PBPK models. The observed tissue/plasma AUC ratios in all major organs correspond well with the reported values of typical mAbs (Shah and Betts, 2013). Also, traditional compartment models have indicated limited tissue distribution of etanercept (Korth-Bradley et al., 2000; Lon et al., 2011). However, more extensive distribution of etanercept was observed in arthritic paws (tissue/plasma AUC is 0.23). The estimated σpaw_dis suggests that etanercept could penetrate well into arthritic paws, which is in agreement with the sparse data of etanercept concentrations reported in synovial fluid in RA patients (Zhou, 2005).
Although nonlinear clearance was observed in the plasma pharmacokinetics of etanercept in CIA rats, the plasma concentration profile of etanercept in CIA rats was not altered significantly in comparison with healthy controls. However, a significant increase in the tissue distribution of etanercept in arthritic paws was observed in CIA rats (tissue/plasma AUC ratios 0.23 versus 0.07 in healthy animals), suggesting that inflammation profoundly altered the tissue distribution of etanercept without causing significant differences in the plasma pharmacokinetics. This is likely to be attributed to increased vascular permeability and fluid retention in interstitial space caused by inflammation in arthritic paws. It has been previously reported that during high-permeability edema in skin caused by bradykinin, lymph flow was increased, and the hydration of matrix and the degree of protein exclusion in the interstitial space changed. All these physiologic alterations led to increased protein concentrations in tissue interstitial space (Bell et al., 1983).
In other tissues, only modest increases of etanercept exposure were observed in the muscle and adipose tissue in CIA rats, and no significant differences were found in the liver, lung, kidney, heart, and spleen between CIA rats and healthy controls. This implies that inflammation may have a minimal impact on these tissues. However, because etanercept concentrations in these tissues were measured as whole tissue concentrations, which are subject to residual blood contamination, the impact of inflammation on tissue exposure of etanercept might be confounded by this factor. We then directly approached collection of ISF from skin using the blister suction technique to avoid blood contamination. Our results indicate that without the confounding effect from residual blood, a 1.5-fold increase in skin interstitial distribution was observed in CIA rats (tissue/plasma AUC ratios 0.43 versus 0.70).
The blister suction technique was previously applied in examining the pharmacokinetics and tissue penetration of small molecule drugs in skin interstitial space (Bernard et al., 1994; Chosidow et al., 1996; Mazzei et al., 2000). For the first time we used this technique to study tissue interstitial distribution of a protein drug. Unlike small molecule drugs, the rate and extent of etanercept distribution in blister fluid is governed by both fluid flow rates during suction and vascular permeability. These contributing factors are well considered with the PBPK modeling approaches.
We first applied a simplified PBPK model to characterize the tissue distribution kinetics of etanercept. The rationale is that 1) convection and lymphatic drainage are assumed to be the dominant pathways for mAb tissue uptake and elimination, and 2) tissue ISF is assumed to be the major extravascular space for mAb tissue distribution (Cao et al., 2013). The simplified PBPK model offers the advantage of assessing the distribution of etanercept in tissue sites of interest, and it yields physiologically relevant parameter estimates in comparison with compartmental models. Also, with reasonable assumptions, the tissue compartment structure is simplified from a full PBPK model. The model can describe the tissue concentrations and can yield reasonable parameter estimates. However, the estimated values of tissue vascular reflection coefficients are highly dependent on the set of values of residual plasma volume used in modeling, which are somewhat variable in the literature. Therefore, to obtain a more precise estimation of tissue permeability, residual blood contamination should be minimized, and if feasible the drug concentrations in tissue interstitial space should be directly sought.
For the quantification of etanercept, the IRDye800CW-labeling approach was applied. Although radiolabeling is traditionally used for mAb quantification in tissues, it requires stringent laboratory regulations. In contrast, IRDye800CW is a near-infrared fluorescent probe providing enhanced signal recovery and high signal-to-noise contrast (Adams et al., 2007). The ease of use, in addition to being cost-effective and requiring no special laboratory restrictions, makes this approach fairly attractive.
Previous studies have assessed the utility of IRDye800 as a quantitative tracer for mAb quantification in plasma and tissues, but controversial observations and conclusions were made. Comparable plasma concentration profiles for up to 24 hours measured by fluorescence and ELISA have been observed for bevacizumab with a high dye/protein labeling ratio (3.5) (Wu et al., 2012). Another study concluded that IRDye800 in a low dye/protein labeling ratio does not significantly alter the pharmacokinetics of cetuximab, which has a relatively short half-life (2.5 days) (Zinn et al., 2015). In contrast, altered mAb pharmacokinetics in plasma and tissues were observed for bevacizumab, cetuximab, and mouse monoclonal IgG1 (8C2), featuring increased plasma elimination and liver uptake (Cohen et al., 2011; Conner et al., 2014). We also observed the apparent increased plasma elimination as well as enhanced liver and kidney uptake of etanercept measured by fluorescence. However, after correction with a decay function as suggested by the in vitro stability study, the plasma concentration profile corresponds well to that measured by ELISA. This indicates that the IRDye800CW-labeling does not significantly alter etanercept plasma pharmacokinetics. The SDS-PAGE of tissue protein extracts showed the presence of increased IRDye800CW-etanercept conjugate fragments in the liver and kidney, which is likely due to the long residual time of IRDye800CW in tissue cells after endocytosis. This is likely to account for the observed enhanced liver and kidney uptake.
In conclusion, along with our previous studies of the plasma pharmacokinetics of etanercept, we have characterized the tissue distribution kinetics of etanercept in arthritic rats. We have shown that etanercept penetrates well into the arthritic paws in CIA rats. For macromolecular drugs, the exposure of drug at the target tissue site is a major determinant of the pharmacodynamic effects. The obtained information for etanercept distribution in joints will help in better understanding its dose–response relationship. The impact of inflammation in the tissue distribution of etanercept was featured. Altered tissue distribution kinetics of etanercept was observed in CIA rats, which we attribute to the increased vascular permeability and fluid retention in the interstitial spaces in the proposed PBPK model. Rodents and humans are considered similar anatomically. The knowledge obtained from the rat will aid in the translation to humans and the prediction of etanercept exposure in the inflamed tissues in RA patients.
Authorship Contributions
Participated in research design: Chen, DuBois, Almon, Jusko.
Conducted experiments: Chen, DuBois.
Performed data analysis: Chen, Jusko.
Wrote or contributed to the writing of the manuscript: Chen, DuBois, Almon, Jusko.
- Received December 23, 2014.
- Accepted April 1, 2015.
This work was supported by the National Institutes of Health National Institute of General Medical Sciences [Grant GM24211] and by the UB Center for Protein Therapeutics.
- UV absorbances
- area under the concentration curve
- collagen-induced arthritic
- D/P ratio
- dye/etanercept ratio
- εetanercept
- molar extinction coefficient of etanercept
- enzyme-linked immunosorbent assay
- IRDye800CW-etanercept
- interstitial fluid
- mAbs
- monoclonal antibodies
- mPBPK
- minimal PBPK
- physiologically based pharmacokinetics
- phosphate-buffered saline
- rheumatoid arthritis
- σ
- reflection coefficient
- TNF-α
- tumor necrosis factor α
- Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics


text:
Technology Appraisal Guidance No. 220
Source: National Institute for Health and Care Excellence
1. Guidance
1.1 Golimumab is recommended as an option for the treatment of active and progressive psoriatic arthritis in adults only if:
- it is used as described for other tumour necrosis factor (TNF) inhibitor treatments in 'Etanercept, infliximab and adalimumab for the treatment of psoriatic arthritis' (NICE technology appraisal guidance 199), and
- the manufacturer provides the 100 mg dose of golimumab at the same cost as the 50 mg dose.
1.2 When using the Psoriatic Arthritis Response Criteria (PsARC; as set out in NICE technology appraisal guidance 199), healthcare professionals should take into account any physical, sensory or learning disabilities, or communication difficulties that could affect a person's responses to components of the PsARC and make any adjustments they consider appropriate.
The guidance shown above constitutes Section 1 of the full document. A copy of the full document and a summary of the evidence is available on the Internet at http://guidance.nice.org.uk/TA220
An abridged version of this guidance (a 'quick reference guide') is also available. Printed copies of the quick reference guide can be obtained by contacting 0845 003 7783 or emailing email@example.com and quoting reference number N2519. It is also available on the Internet at http://guidance.nice.org.uk/TA220/quickrefguide/pdf/English
This guidance represents the view of the Institute which was arrived at after careful consideration of the available evidence. Health professionals are expected to fully take it into account when exercising their clinical judgement. This guidance does not, however, override the individual responsibility of health professionals to make appropriate decisions in the circumstances of the individual patient, in consultation with the patient and/or guardian or carer.
© Copyright National Institute for Health and Care Excellence. All rights reserved. This material may be freely reproduced for educational and not for profit purposes within the NHS. No reproduction by or for commercial organisations is permitted without the express written permission of the Institute.
Enquiries concerning the guidance should be addressed to: National Institute for Health and Care Excellence, MidCity Place, 71 High Holborn, London WC1V 6NA. email: email@example.com
Golimumab for the Treatment of Psoriatic Arthritis
Issue Date: April 2011

subset: commoncrawl


text:
Liverpool may have to break their club record fee again if they want to land AS Roma number one next summer.
The Reds are currently battling with Real Madrid for the signature of Allisson after their goalkeepers have failed to show they are world class.
Jurge Klopp has had to rotate Loris Karius and Simon Mignolet throughout the first half of the season. Although he has made Karius the permanent first choice this year, the German has still not convinced watchers that he can lead them to the Premier League title, this uncertainty has continued to link the Reds with other goalies.
They were reportedly keen on signing AS Roma's Allisson last month and remain keen ahead of the next transfer window.
However, Calciomercato reports that the Reds now face the prospect of paying £76 million for the Brazilian. Per the reports Roma are preparing to offer Allisson a new deal worth £4 million per season and increase his release clause to £76 million.
This figure may appear too much for Liverpool to pay, but they face the prospect of missing out on signing him after Real Madrid joined the race for his signature and Los Blancos would have no problem paying close to that figure.


text:
The seal of the community of Wienhausen.
wax; wax impression of the convent's seal
seals of the abbess and provost; Wienhausen, seal of the abbessWienhausen, seal of the provost
The inscription on the seal reads "Sigillum sancte Marie in Winhusen." The virgin Mary is depicted in the center, standing and holding the figure of the Christ child. To her right stand two figures bearing candles (?). To her left appear seven figures grouped together, perhaps a reference to the Cistercian nuns of the community or the seven wise virgins.
Klosterarchiv Wienhausen,Wienhausen
Abbess Renate von Randow; Wolfgang Brandis


text:
IMTS 2020 September 14th – 19th, 2020
Come visit our booth
We are going International Come visit us September 16-21 2019.
Come visit our booth! Show Information Date: September 12-14, 2017 | Los Angeles Convention Center, Los Angeles, CA
Come visit our booth! Show Information IMTS 2018 will be the 32nd edition of the premier manufacturing technology show in North America. IMTS 2016 hosted the highest number of exhibiting companies ever (2,407) and was the third largest in number of registrations (115,612) and in net square feet of exhibit space (1,370,256) at the McCormick Place


text:
(By Pat 'Riot' Whitaker, Senior Writer/Journalist, RiffRelevant.com)
WE ARE SENTINELS is the new project of former Iced Earth / current Ashes Of Ares vocalist Matt Barlow and keyboardist and pianist Jonah Weingarten (Teramaze, Pyramaze).
Today, they are sharing further information about the band's endeavors, including that their self-titled début album will be released on July 6th via SAOL. WE ARE SENTINEL have shared a pair of tracks from the album with a lyric video for "Battle In Winter", an official video for "Kingdom In Winter", and both are streaming below.
Pre-order for the self-titled début from WE ARE SENTINELS is active now at these links:
U.S. pre-order HERE / iTUNES pre-order HERE / Amazon HERE
Physical Album (Germany) pre-order HERE


text:
Due to annual maintenance on our servers, you may find some parts of our website pages disturbed a little. We are sorry for the inconvenience.
With a commitment of excellence and quality, RDU Delivered will surely deliver the most proficient moving services in Charlotte NC. We have proudly been the first choice of our satisfied customers for any type of move since 2013. With every passing year, our company has grown strong and more professional as a team. Our movers are all that you need to move with a class.
We do not claim what we cannot do. Honesty and integrity are the foundation of our popularity in the industry. You are our priority and we will go an extra mile to make your satisfaction certain. Our company is indulged so much in the perfection of the project that our movers are known for exceeding the expectations of the clients. Moving, hauling, and relocations are hard to do the job when you have no idea how it is done. With our top-notch skills RDU Delivered will serve you with the best long distance moving, long distance hauling, long distance relocation, long distance commercial moving, top-rated interstate moving, interstate hauling, interstate relocation, interstate commercial moving, affordable cross country moving, cross country hauling, cross country relocation, cross country commercial move, white glove moving, corporate moving, and corporate relocation in Charlotte NC.
We are the best version of ourselves because we improve and grow every day and that is what makes us stand out from the other moving services. We offer free estimates and our rates are also affordable. Our movers will set up a proper schedule with you and visit your property to get an exact idea of the time and effort needed to make these following moves successful for you in Charlotte NC:
When it comes to a long distance relocation or hauling, especially in a long distance commercial moving, we are among the best and most affordable long distance moving companies in Charlotte NC. We provide a friendly and reliable service as our movers are fully licensed and insured.
Searching for the top-rated moving companies will easily lead you to us for getting the best interstate commercial moving or hauling experience in Charlotte NC. Your satisfaction is our guarantee. Our movers are equipped with licensed trucks and all the latest moving equipment.
The hardest part of making a cross-country move is to trust a company for that. When you are relocating your office or commercial unit to a new country, you need top-rated and affordable cross country moving companies that you can count on for the safe and secure move.
During your corporate moving or relocations, you will get to know how the real white glove moving is done. Our company will serve you with luxurious and superior moving service that will turn a hectic task into a comfort for you in Charlotte NC.
For details, Please feel free to call us at 919-758-9462


text:
The crew over at Menstrual Monday will be hosting their 2nd annual ladies-only Girls of Summer alleycat tomorrow and it should be awesome. Just look at the sponsor list! Word has it, it's over $3000 in prizes. Registration starts at 2pm at the I-5 Colonnade (race at 3pm) but show up early if you'd like your bike looked over by the crew from Back Alley Bike Repair; they'll be there starting at 1:30pm.
Go Means Go!
Alaskan life on two wheels.


text:
The Answer to Left Fear. Fast, Effective and Guaranteed
Feelings of dread? Overwhelming thoughts? Rapid heartbeat? Tunnel Vision? Or worse? Since 2002 we have worked with countless folks from all walks of life and we have yet to come across someone with a case of left fear that couldn't be overcome.
If you are now ready to have this over and done with, here's what you can expect from us:
- A noticeable improvement, often a major one, in less than 24 hours
- Complete confidentiality and privacy
- Clients in over 70 countries around the world
How to Get Started
We offer two programs for left fear. You can choose to experience the Home Study program (going at your own pace on your own time), or choose to work one-on-one with a highly-qualified practitioner:
Home-Study Program: Vanquish Fear & Anxiety
CD's and Workbook $147 or Download for $137
• Start Instantly with Online 'Quick-Start'
• Go at Your Own Speed in the Privacy of Your Home
One-on-One 'VIP' Program
Custom Program Developed for you by Your Expert Practitioner
• Confidential Sessions with Board-Certified Specialist
• Sophisticated Suite of Techniques to Remove Underlying Fear
How This System Works
You've got to 're-wire' your unconscious mind – and with our help its easier than you may think. Your rational mind knows that your Left Fear is illogical. Yet the fear perseveres only because your subconscious has 'linked' the thought of left-sidedness to a bunch of strong negative feelings – which get brought up automatically every time.
Until now, you just haven't had the techniques for re-programming those connections. We're here to give you give you what you need to take back control. Success is on the way – all you need to do is get started:
- Are you ready to speak with a Board-Certified Specialist? Contact Us Now Contact Us Now >
What You Need to Know About Treatments and Cures
Here's the thing, our system does not treat or cure your fear. Modifying your awareness is a technique you have to learn to master for yourself. We are here purely to teach, and to hold your hand as you walk through the procedure that enables you to make it happen.
The process educates you with exact steps on how you gain self-confidence, calm and happiness, as well as proven methods to overcome anger, sadness, fear, hurt, guilt and anxiety. You have dealt with these emotions haunting you long enough. It is time for you to stop that cycle of negative thoughts.
Left Fear Risk Factors and Triggers
In most cases, Left Fear (although not always) is triggered by an extremely negative encounter from the past. In some cases, it appears the brain has produced the fear with no instigation. By exposing the root cause, you will be able to remedy the problem by easily exchanging harmful triggers. Individuals utmost at risk are:
• Those who have a general predisposition for fear and anxiety
• Someone considered to be 'easily upset'
• People suffering from adrenal insufficiency
Any of those portray you? You can see testimonials from individuals who know our our capability to make a difference.
Would you like to read about us in the media instead?
Diagnosing Your Phobia
Use our 2 minute Left Fear Online Analysis to discover the magnitude of the problem for you, but it is pretty simple. Now is the point in time to take control, if the fear is considerably taking over your life, in a negative way.
- Does the thought of Left-sidedness make you nauseous?
- Does it trigger a dry mouth and clammy hands?
- Does your heart begin to race?
- Do your legs feel as if they will give way?
Those are just a few of the symptoms. Click here to see more symptoms of left fear.
We address the symptoms, but we uncover the source, to eliminate them both. This is what CTRN is all about
Drugs & Medication
We feel drugs and medication for Left Fear aren't the best approach, but make sure you always follow a doctor's advice when taking or stopping any prescription. Pills might help you cope with your Left Fear but only in the short term – they will not, in reality, help you eliminate it forever.
No drug has ever been developed specifically for Left Fear, by the way – When looking for an answer, maybe you should think about possible side effects before turning to prescription drugs.
The good news is with our system you will vanquish your Left Fear, without drugs..
Next Action
- Learn about The VIP One on One Program:
The VIP Program for Left Fear
- Discover The Home Study Program:
Vanquish Fear & Anxiety for Left Fear
Or learn more about Left Fear:


text:
Horror films are poignant, cultural commentaries, reflecting our fears back at us. Yes, you may want to sit back, turn on a scary movie, turn off your brain, and just watch giant mutated animals fight each other, but you can't ignore what they're saying about their cultural contexts. Take Ishirō Honda's 1954 classic, Godzilla. At face value, it is a silly movie about a giant lizard stomping on Tokyo while crowds point and scream, "Gojira!" However, it's more than just an old monster movie — it is a cutting reaction to the 1945 bombing of Hiroshima and Nagasaki and a condemnation of nuclear power. Godzilla is literally awoken by underwater hydrogen bomb testing. But even more, Godzilla's destruction is reminiscent of these bombings. As he crushes buildings and demolishes cities with his nuclear breath, images of devastated cities are conjured up. Characters in the 1954 film even reference the bombings when discussing their fears of the giant lizard. While these films are weird and wacky, they also serve as a reminder of the atrocities Japan has suffered at the hands of Western society.
The puppetry is ridiculous and writing can be laughable, but there's no doubting Godzilla's influence on the monster movie genre. These five films are the best Godzilla movies Criterion has to offer, from their message to outright monster-fighting hilarity.
Godzilla (1954) dir. Ishirō Honda
Nothing can beat the original Godzilla. Hydrogen bomb testing awakens an ancient beast from his aquatic slumber. He begins to destroy Tokyo and the government rushes to find a solution. The only way to defeat the monster is to use the "Oxygen Destroyer" created by Dr. Daisuke Serizawa. However, Serizawa can't live with himself after causing such destruction and sacrifices himself to stop the monster.
While many of the following films are silly, this first installment is much bleaker. It doesn't shy away from more graphic images of those injured by Godzilla, and talks explicitly about the history of the hydrogen bombs. His destruction isn't seen as funny; it is devastating and traumatizing. The film is Japan coping with trauma by disguising it as a monster movie.
Mothra vs Godzilla (1964) dir. Ishirō Honda
Mothra vs Godzilla introduces my favorite opponent, Mothra. The film begins with a mysterious giant egg spotted in the ocean. As it washes up on shore, it becomes the center of the country's attention. Scientists want to study it, citizens want to touch it, and businessmen want to buy it. After a rather generous offer, the egg is purchased and put on display. However, these are Mothra's eggs, and must be protected. Once Godzilla arrives and Mothra is the only hope, it's a race to save the eggs and hatch them before it's too late.
Mothra is Godzilla's best opponent for two reasons. First, she is one of the only female monsters in the Godzilla canon. Gender isn't relevant for giant monsters, but since a gender is constantly assigned to them, might as well get excited for the one female character. Second, she is always recruited to help fight against Godzilla, or at least talk reason into him. She is the only monster who is partners with humans instead of trying to destroy them.
Destroy All Monsters (1968) dir. Ishirō Honda
Somehow Japan has gathered Godzilla and most of his enemies onto one island where they live in apparent harmony. However, aliens arrive on Earth and release these creatures from the island, sending them to cause destruction around the world. Then, the dreaded King Ghidorah appears and the monsters reunite to defeat him.
This movie is high on my list solely for its ending. There's a lack of plot and what plot there is moves at a snail's pace, which is surprising due to the alien invasion. Once all of the monsters start attacking Ghidorah, it's worth it.
Ghidorah, the Three-Headed Monster (1964) dir. Ishirō Honda
This film is the introduction to Godzilla's nemesis, Ghidorah, a golden three-headed dragon from space. He destroyed the civilization on Venus and now he has come to Earth to do the same. To have any chance at defeating him, Godzilla must partner with Mothra and Rodan to fight the alien beast. Meanwhile, a citizen of Venus takes over a human's body to warn the humans of the imminent destruction. On top of that are assassination attempts towards a princess, a struggling journalist, and a bodyguard cop. Insane? Yes. Fun? Of course.
In Ghidorah, Mothra makes another appearance, this time acting as the voice of reason to a childish Godzilla and Rodan. It is absolutely ridiculous to watch a moth talk to a giant lizard and giant bird creature and try to convince them to fight a three-headed dragon from space. But this ridiculousness is what makes the film so great. Plus, we get to see Godzilla behave like a petulant child and kick rocks at his enemies. The biggest issue with this film is the middle portion where Honda attempts human character development in a film about a giant alien dragon. Needless to say, we don't need that exposition.
Godzilla vs. Mechagodzilla (1974) dir. Jun Fukuda
What's better than one giant radioactive lizard? Two of them, but one is actually a robot version built by aliens who want to take over Earth. This group of aliens has been watching Godzilla and building a replica of him to conquer Earth. However, Godzilla has some help from King Caesar, an ancient deity awoken to protect humanity.
While brief, Godzilla vs. Mechagodzilla discusses issues with Japanese colonialism, particularly on the island of Okinawa. The Japanese took over the island in the late-19th century and attempted to eradicate their culture. This film reflects this history by including the Azumi tribe. A relic is stolen from them by archaeologists, which is said to help protect their village. Only when it is returned can they awaken King Caesar who can help save Japan. The tribe's leader is also portrayed using slapstick, exaggerating his movements and covering him in stereotypical old man makeup. He is immediately established as Other from the rest of the Japanese character. In a film about fighting a robot monster, there is cultural commentary woven throughout addressing Japan's troubled past.


text:
Consider the everyday uses of the veil: some cultures use it to hide the faces of women, some use it to tantalize, thinly draping the body so that only an outline can be seen The veil is a transition, as when a groom lifts the veil of the bride to take her from a single woman into a new life as a wife, or between one room and another.
These mundane representations serve to symbolize the implications of the mystical and occult, where the veil stands for limitations of awareness that prevent human beings from directly perceiving divine realities. Exemplifying this concept is the famous veil from ancient Israel, the Veil of the Sanctuary, said to separate the room where the Ark of the Covenant, a vessel of God, was kept from the rest of King Solomon's temple. According to the Book of Exodus, it was purple, scarlet, and blue. These colors are symbolic of the crown chakra, allowing divine connection to Spirit (purple), the root chakra, the base of the source or Kundalini energy (scarlet), and the throat chakra, the center of communication (blue). The Books of Mark and Luke indicate that the Veil of the Sanctuary tore when Jesus was crucified. The barrier between the physical and the spirit planes which separated God from humanity had been broken, the boundary had been transcended, and so the Veil tore.
In mythology, the goddess Ishtar is allowed to pass from the world of the living to the underworld, but must remove an article of clothing at each of the seven gates, this being one possible origin of the story for the Dance of the Seven Veils. The gradual peeling away is symbolic of the necessary steps to reveal one's true nature, a shedding of external layers to uncover the essence of the individual to have a clear understanding of the self and to be closer to Spirit. It represents the transition essential to transform from one state to the next as one grows personally and spiritually.
The Veil as the filmy barrier between the world of the physical and that of the spirit varies in density depending on the time of year and the individual's ability to see the spirit world. For those with psychic ability, the view is still shrouded in fog, yet accessible a good amount of the time. For those without the gift of second sight, the Veil is thick and impenetrable. The spirit world is not available to one such as this. During certain times of the year – autumn and Halloween in particular – the Veil is thinnest and is the best time for divination. Those who are able to access the Veil have experienced it as a place of silence, calm, and peace. All that is outside of the Veil is motion, noise, and desire. To be in the Veil is to be both separate from the physical world and a part of the oneness of Spirit.
Wearing a veil over our true selves blurs the truth of who we are. If the veil is intentionally placed, it can be used to control perception by others to avoid awareness of your actions and intentions before it is time for them to be revealed. The veil can also unintentionally serve to make communication difficult by preventing others from understanding what you are doing and why.
A veil that grows over time as a result of misconceptions, lack of will, and refusal to see one's self clearly hinders accurate assessment of the self and stunts the individual's personal and spiritual growth. The veil is also the transitional screen between the light and the shadow self. Everyone has a shadow self. The dark side is only thinly veiled in some and thickly veiled in others. It provides a balance within us and needs to be acknowledged and addressed in a healthy way. When out of balance, these aspects of dark and light present in identifiable ways.
Those tending toward the shadow self most likely cloak themselves in a thick veil to hide their dark intentions in order to be more effective at manipulating others. Those who tend toward the light do not feel a strong need to hide behind a veil, as their projected self is sincere. At the same time, these individuals need to acknowledge the existence of their shadow self rather than ignore, suppress, or let it out only under the most stressful circumstances. These individuals also may be prone to judging others. Without awareness and recognition of the darkness we all possess, no matter how minimal, a tendency toward anxiety and/or depression may develop. Ultimately, the lack of self-knowledge is the most devastating result.
The veil masks the characteristics of those who are self-serving and seek to survive in a way that deceives others. The veil also comes down when there is an emotional trauma and the psyche seeks to deal with the powerful emotions the situation conjures. This energy pulls the curtain down on the person's natural way of being and clouds the mind. It tarnishes the auric field and dulls the senses. In a way, this is a protective mechanism for the traumatized person to be able to withstand the physical and emotional injury. At the same time, to look through the veil of sorrow or grief affords a spotty view of the world. Clarity comes when the emotional disruption is once again balanced.
The Veil is an etheric shield that is draped across our energy bodies, as well as between the world of life and the spirit realm. The Veil clouds, distorts, and hides, yet when used to best advantage, the Veil can provide insight, protection, and concealment.
As you consider this information ask yourself the following:
Your thoughts and feedback are welcomed and encouraged!
Many blessings, Diane Wing
Diane Wing is an author and owner of Vibrant Concepts, LLC. She combines practical business experience with metaphysical concepts to help her clients better understand and trust themselves, make better decisions, and be more tolerant of change. Visit Diane at www.VibrantConcepts.com; email@example.com.


text:
Dyan Branstetter | February 2017
Using the "Evidence For, Evidence Against" Strategy
"Evidence For, Evidence Against" is a teaching strategy that helps students "close read" a piece of text. Under the Common Core Standards' definition for text, literature, music, visual art, dance, and drama apply. This allows us to successfully apply the teaching strategy across our Arts subjects. "Evidence For, Evidence Against" compares to a rigorous true and false activity, requiring students to share evidence to support or argue a statement. This forces students to go back to the text and examine it closely to respond. There is no way to do this without thinking deeply about the text itself, allowing students to make discoveries they may have otherwise overlooked.
How to Implement "Evidence For, Evidence Against"
To use this teaching strategy, students are presented with a three-column chart that includes statements in the middle column. This can be used after reading or during the reading experience. The statements are pulled from the text, however, some are true statements, and some are not. Students determine if it is true or false, and then provide the text evidence to prove it correct or incorrect. For example, take a look at an excerpt from the text Lon Po Po: A Little Red Riding Story by Ed Young. Then, take a look at a simple example on the Evidence Chart excerpt with the student answer.
This example is directly related to the words the author uses, but the same concept applies when using the strategy to prove the theme, pointing out the author's craft, or with nonfiction guide students to analyze text structure. It also requires them to know and use tier 2 vocabulary.
Using "Evidence For, Evidence Against" with the Arts
When using this teaching strategy with the Arts, the same guidelines apply. To follow the way we used it with literature, students can find Evidence For or Evidence Against visual art by analyzing the content of the art. Create a true or false statement for the chart. Statements, of course, will be different depending on your grade level and how much background your students have on the topic and the art form. This can apply to music, dance, and theater as well.
Take a look at the example for the piece The Buffalo Hunt [No. 29] by Charles M. Russell. Students can explore this artwork (as well as many others) virtually through Google's Arts and Culture "Zoom In" section, found here.
Introducing the Teaching Strategy to Students
When I use this teaching strategy with students, we first complete it as a whole group so that I can demonstrate the amount and type of evidence we need. Students sometimes get confused on their own, thinking that they need to fill in all of the boxes on the chart. After the initial instruction, I almost always have them use this strategy with the text and a partner. If they are without the text, as some students think they can rely on their memory, it is impossible to cite the evidence at a specific level. Allowing students to work with a partner can help students bounce ideas off one another and they are able to work together as a team to find the evidence they need.
After The Strategy
We follow up by discussing answers as a class. This tends to get heated, but in a good way. Students share their evidence, and if the statement is challenging enough, other students tend to disagree, basing their argument on different evidence in the text. This type of disagreement is a great way for the group to grapple with the text itself to determine what the author or artist means. It takes a bit of training to help students "argue" in a beneficial way, but they have a lot of fun when the discussions become passionate.
I love hearing a student explode with a statement like, "No, no- I disagree! On page 16, the author says …., so that CAN'T be true." I've worked to create a culture where students can disagree with each other (and with me!) provided they have the evidence to support it. Not only does it allow for great higher-level discussions, but it gives students purpose for their learning while getting them back into the text. It also helps them develop the necessary social skills for disagreements.
"Evidence For, Evidence Against" Extensions
After students master the strategy, I have them create the statements. I typically do this by drawing the three-column chart on a piece of chart paper. As they think of a tricky true or false statement, they add it to the middle column. Other student groups can write on the chart to prove or disprove the statement with evidence.
Are you interested in trying this strategy with your students? Check out the resources below. You'll find a full example for the story Lon Po Po, created by a colleague of mine, without student answers. In addition, you can download an editable blank version for you to try with any piece of text or art.


text:
Stress Relief mp3
Stress Relief mp3
When stress gets too much it can affect us in many ways from insomnia to high blood pressure and stress headaches to full blown migraine. This stress relief mp3 is specifically designed to help reduce stress-related symptoms, returning you to a deeper sense of well-being.
It can take as little as 10 minutes a day. Go on, treat yourself!


text:
Emily Benfer
- Visiting Associate Clinical Professor of Law
Emily A. Benfer is a visiting associate clinical professor of law. As a well-known authority in the field of health justice, Benfer is recognized for a myriad of social justice and scholarly accomplishments, and a commitment to preparing the next generation of advocates. Her area of clinic practice and research includes health equity and social justice, interprofessional education and collaboration, environmental and racial justice, and medical-legal partnership.
Prior to joining the faculty to direct the Health Justice Advocacy Clinic, Benfer was a Visiting Distinguished Scholar and Senior Fellow at Yale Law School Solomon Center for Health Law & Policy. Between 2010-2017, Benfer served as a Clinical Professor of Law at Loyola University Chicago School of Law and Loyola University Chicago Stritch School of Medicine Department of Public Health where she founded and directed the Health Justice Project, a fully integrated medical-legal partnership at Erie Family Health Center in which providers, lawyers, and law students collaborated to resolve the social determinants of poor health for patients. The Health Justice Project received the CLEA Award for Excellence in a Public Interest Case or Project for student leadership in an interprofessional advocacy campaign to protect the four million children in federally assisted housing nationwide from lead poisoning. In addition, the Health Justice Project received the Outstanding Medical-Legal Partnership Award from the National Center for Medical-Legal Partnership.
Before founding the Health Justice Project, Professor Benfer served as a teaching fellow and supervising attorney in the Federal Legislation & Administrative Clinic at Georgetown Law Center. In practice, she was a legislative lawyer in the successful efforts to pass the ADA Amendments Act; to require education assistance for homeless children and youth in Washington, DC; and to align the Illinois definition of lead poisoning with the Centers for Disease Control and Prevention standard. She has engaged in direct representation, class action litigation, grassroots organizing, creative advocacy strategies, and federal and state policy reform in multiple areas of public interest law, including homelessness, lead poisoning prevention, special education, housing, health, environmental justice, disability, and public benefits at non-profit organizations and a public interest law firm. Benfer has testified before Congress and appeared in numerous media outlets. She served as an Equal Justice Works Fellow and a Peace Corps volunteer.
She was named a Legal Freedom Fighter by Rocket Matter and one of Chicago's Top 40 Lawyers Under 40 by the National Law Journal. She has received numerous commendations for her commitment to health equity and social justice, including the American Public Health Association David P. Rall Award for Advocacy, Health Innovator Award and the inaugural Schweitzer Leadership Award. In addition, Professor Benfer was named Chicago Person of the Year (group award) for her pro bono work to organize attorneys at O'Hare International Airport in response to the travel ban. Benfer earned her LLM from Georgetown Law Center and JD from Indiana University Robert H. McKinney School of Law. She received a Certificate in Social Justice and a Certificate in Nonprofit Leadership and Development from Loyola University Chicago.


text:
Locals We Love | May Edition
Have you checked out one of our monthly networking and social events? Every event is so unique and individualized which makes them so much fun! The events are hosted at various venues around the Fort Collins and Northern Colorado area along with spotlighting local businesses and building relationships and community with one another. Each event is similar yet so different and May's was exactly that. What a fabulous evening filled with building connections, delicious donuts from Peace, Love and Little Donuts and craft brews from Gilded Goat Brewing.
We missed those of you who could not make it and hope to see you on June 14th at Bad Daddy's Burger Bar Ladies Networking + Social- RSVP, here. Check out the photos captured by EagleIMedia from this fabulous evening.
In the meantime, I wanted to share with you all the businesses that were spotlighted throughout this event. If you are in need of their services or products, don't hesitate to contact them. They would love to connect with you. So, here are the Locals We Love: May Edition.
A HUGE thank you to the Gilded Goat Brewing Company in Fort Collins for hosting our event this month. What a unique and beautiful space. It is so awesome to be able to look directly into the brewhouse from up above. We will be back.
Maxline Brewing Social + Networking
Are you coming to the Ladies Night Out event at Maxline Brewing on October 19th from 5:30-8:30pm? Check out the Facebook event page and don't forget to register for your FREE ticket at eventbrite
here's the inside scoop
We are one week away from this awesome event. ONE WEEK! We'll open the doors at 5:30, try to get there early, because our first fifty guests at Maxline Brewing will be given a ticket redeemable for one of our SWAG Bags! Yay!
Maxline has beer, Kombucha, and other thirst quenchers. Farm Fusion has a delicious menu they will be offering throughout the evening, so bring your appetite and Something Sweet- Fort Collins will have a sweet display of treats to taste.
I'll be giving away a gift certificate for TWO FREE meals from Farm Fusion at 6:30pm and giving away a FREE growler and growler fill from Maxline Brewing at 7:00 PM. You can enter these giveaways at registration, just fill out our cards with your name and email or drop your business card in to be entered. Those who have RSVP'd on Eventbrite and show up will be entered into the drawing for a Family Fall Harvest pack to the Harvest Farm Fall Festival.
After the giveaway at 6:30 PM, I'll announce that it's time to collect the SWAG Bags! Bring me your ticket back to claim your bag!
A few of our featured vendors will be running giveaways too! They are in charge of their own giveaways, and you can find out about them by visiting their tables! You won't want to miss out on these!
I've lined up some great vendors for you guys this time! Take a second and check them all out!
Amanda and Erin with Rodan + Fields
Janna Fiala with Keep Collective
Tatum Stoner with LulaRoe
Shannon Fabian with Chloe and Isabel Jewelry
Alisha and Valerie with Repurposed Petals
Michelle Weidenbach with Le-Vel
Something Sweet- Fort Collins
Farm Fusion
Our event will be captured by the awesome Jackie Clarkson Photography. She will be venturing around the event and taking some candid shots as well as group photos. So if you see her, don't be afraid to ask her to capture your group of girlfriends. Photos will be posted on facebook post the event. Check out her work at Jackie Clarkson Photography
Charity Partnership
I am so excited to share that we are partnering with U COUNT Campaign for this event. To support U COUNT, please bring travel sized toiletries- shampoo, conditioner, face wash, body wash, deodorant, toothbrush, toothpaste, etc.
The U COUNT Campaign is a team effort to bring hope and freedom to victims of sexual exploitation by:
- Raising funds and working in partnership with Project Rescue to provide safe residential and vocational training environments.
- Economically empowering at risk women and girls, as well as former victims of sexual slavery, through the marketing of the products they make.
- Supporting local and global initiatives that provide preventiontrainings and community organization working on behalf of trafficking survivors.
Hi! I am Lisa Downer, married mother of 3 girls and a 20+ year resident of NOCO! I was introduced to HITC in January 2018 and fell in love with the concept of not only networking with local women in business but promoting/sharing them and what they do!
We have such a unique mission and it is right inline with my personal business passions! Come be a part of our amazing group of supportive women and learn how to build your Tribe right here in your own market! with me!
June 2020


text:
Celeste is a renowned opera diva who retired early for the man she loved to live on a crumbling and beautiful estate in the heart of a rainforest in Far North Queensland. Ten years after the tragic death of her husband, Mateos, in a boating accident, Celeste is set to return to the stage for her final performance. Her husband's son Jack, still haunted by the past, arrives amidst the preparations for the performance and finds Celeste is as he remembered – beautiful, intoxicating and dangerous. Celeste wants Jack to stay at the estate, but needs him to perform one last request.
Views: 551
Director: Ben Hackworth, Peter McLennan
Actors: Bryan Probets, Emm Wiseman, Jason Klarwein, Kai Lewins, Kevin Spink, Nadine Garner, Odessa Young, Radha Mitchell, Ron Kelly, Thomas Cocquerel
Country: Australia
Duration: 105
Quality: 4K
Release: 2018
IMDb: 6.1
Leave a Reply
You must be logged in to post a comment.


text: My name is Taylor Elliott, I am a senior at Woodstock High School. In my free time, I usually watch whatever current Netflix show I am on, or I love to go shopping. I love hanging out with my friends, even if it is just watching movies on a Saturday night. I will be continuing my education at Kennesaw State University in the fall, and I will be majoring in Biology. I have so any great memories from Woodstock, but my favorites are probably at lunch, or at the Friday night football games. There was never a dull moment in either of those. My favorite article was probably my first one. Lexi and I always did "Senior of the Month," and for our first one we did one of the star football players. It was just so interesting seeing another side to that jock that everyone knew. These articles were always really fun because, we have gotten to meet and get to know so many new people. My favorite memory of Journalism class was getting to chat with all the new friends I made in this class, not to mention Armando always kept it interesting.


text:
The Duke and Duchess of Sussex will soon be heading to Africa for their first official tour with son Archie, which can only mean one thing: Meghan will be taking an enviable wardrobe with her.
A royal tour means countless wardrobe changes and if the couple's previous engagements are anything to go by, the Duchess will step out in a number of designer labels and luxury accessories.
One such number is the elegant, two-tone Roksanda dress the new mum wore to visit the Macarthur Girls High School during her 18-day Australian tour.
Meghan accessorised her navy pleated dress with nude Stuart Weitzman Legend pumps, Birks blue topaz earrings, and a Birks white-gold flex bracelet, while her hair was styled in a half-up style with loose curls.
The Athena gown retails for a hefty £850 and while the blue has completely sold out, it is still available in a beautiful pink colourway.
READ MORE: Everything we know about Meghan Markle's Smartworks womenswear line
But if you don't have that much to spend, Hobbs London has a near-identical dress for a fraction of the price.
The Helenora Dress retails at £89, but is currently on sale for £59.
Hobbs London Helenora Dress | £59 (WAS £89) | Shop now
The description reads: "Our Helenora dress nips in at the waist for a confident, hourglass silhouette while the flared skirt only adds to the femininity. The boat neckline brings an air of elegance while the woven colour block hem adds interest."
While the Hobbs version has short sleeves, the navy dress features a vibrant two-tone design exactly like the luxury Roksanda ensemble, which can be styled with a pair of flats or court shoes to take the dress from day to night.
The gown is currently available in sizes 10-16.
The dress has already received plenty of praise online, with one shopper describing the gown as "very flattering".
READ MORE: Meghan Markle's (surprisingly affordable) £45 sunglasses are back in stock
One wrote: "This is a lovely dress, great quality materials, well cut and very flattering. The length is mid calf on me (5'7) and looks great with heels or flats.
"Ideal for any "smart casual" occasion. The bright blue stripe at the base is made from a different material (body is a thick stretch jersey) but that doesn't detract, although it will crease unlike the rest. Love it!"
As with anything Meghan wears, we expect this number will sell out in no time, so you ought to be quick.
The editors at Yahoo UK are committed to finding you the best products at the best prices. At times, Yahoo UK may receive a share from purchases made via links on this page.


text:
So, who are Manchester's biggest bands of all time? We've covered the 60's, the 70's and of course the cities "Madchester" era during the '80s. Check our top 10 below. As always, if missed someone, let us know.
Formed in 1991, Oasis are purveyors of no-nonsense, in your face anthemic, British guitar rock. Liam and Noel Gallagher both hail from the Manchester suburb of Burnage. Liam had joined a local band The Rain(featuring eventual Oasis members Paul McGuigan and Bonehead), soon joined the band and the outfit became the classic Oasis line-up. Signing to indie label Creation in 1993, the band released the huge selling albums Definitely Maybe(1994) and (What's The Story) Morning Glory? (1995). Oasis has sold over 75 million records worldwide, and have had eight UK number-one singles and eight UK number-one albums. They were listed in the Guinness World Records book in 2010 for "Longest Top 10 UK Chart Run by a Group" after an unprecedented run of 22 top 10 hits in the UK.The band also holds the Guinness World Record for the most successful act in the UK between the years 1995 and 2005, spending 765 weeks in the top 75 singles and albums charts. After releasing its seventh album, Dig Out Your Soul, in 2008, the band split.
Formed in Manchester in 1982, Smiths singer Morrissey looked as if he would make a living as a writer or poet, but pop stardom was always on the horizon and upon meeting guitarist and songwriter Johnny Marr, the foundations were built for one of Britain's greatest pop bands. After the addition of bassist Andy Rourke and drummer Mike Joyce, the Smiths created four classic albums and over a dozen 'perfect' pop singles. Critics have called them one of the most important bands to emerge from the British independent music scene of the 1980s. NME named the Smiths the "most influential artist ever" in a 2002 poll, and in 2003, three of the band's albums appeared on Rolling Stone's list of the 500 Greatest Albums of All Time. Following their split in 1987, rumors of a reunion continue to circulate to this day.
The Hollies became one of the leading British groups of the 1960s. If you need proof, how about 231 weeks on the UK singles charts during the 1960s. Formed by Allan Clarke and Graham Nash in 1962, Nash ended up leaving the group in 1968 to form the super-group Crosby, Stills & Nash. The Hollies could certainly write pop songs with a deeper meaning, and their track 'Bus Stop' is firmly rooted in the everyday normality of life around the small towns surrounding Manchester. Decades later The Smiths and Oasis would be writing about the same subject matter. Over five decades the band has had at least 60 singles or EPs and 26 albums charting somewhere in the world. The Hollies had over 30 charting singles on the UK Singles Chart, and 22 on the Billboard Hot 100, with major hits on both sides of the Atlantic that included "Just One Look", "Look Through Any Window", "I Can't Let Go," "Stop Stop Stop", "On a Carousel", "Carrie Anne", "Jennifer Eccles", and later "He Ain't Heavy, He's My Brother", "Long Cool Woman in a Black Dress", and "The Air That I Breathe" The Hollies were inducted into the Rock and Roll Hall of Fame in 2010.
Joy Division was formed in Salford, Greater Manchester, in 1976. The band consisted of singer Ian Curtis, guitarist, and keyboardist Bernard Sumner, bassist Peter Hook and drummer Stephen Morris. Sumner and Hook formed the band after attending a Sex Pistols concert and originally started out life as young punks until their love of Bowie, The Velvet Underground, and synth-propelled prog rock took them into a darker direction. Their self-released 1978 debut EP An Ideal for Living drew the attention of the Manchester television personality Tony Wilson, who signed them to his independent label Factory Records, and their debut album Unknown Pleasures was released in 1979. Sadly, by the time the follow-up Closer was released, Curtis has died: he committed suicide in May 1980. Joy division as we knew it was over, but the rest of the band vowed to carry on…
Ian Brown and John Squire met at Altrincham Grammar School and formed The Stone Roses Manchester in 1983. Known as one of the founders and pioneers of the 'Madchester' movement in the late 1980s and early 1990s, the band's lineup consists of vocalist Ian Brown, guitarist John Squire, bassist Mani, and drummer Reni. Their debut album, The Stone Roses was released in 1989 and the band garnered critical acclaim overnight, with many critics regarding it as one of the greatest British albums ever recorded. High praise indeed. The album was a mix of classic indie and psychedelia, and it was 5 long years before a follow-up arrived: titled The Second Coming. The band called it quits in 1996, but in 2011 they announced comeback shows and new material. Fast forward to the 24thof June 2017, the Stone Roses played at Hampden Park in Glasgow. During the performance, Ian Brown addressed the crowd with the statement: "Don't be sad that it's over, be happy that it happened," leading many to speculate that the performance would be their final concert.
Easily one of City's most-loved bands, Elbow were formed by Guy Garvey and Craig Potter after they met at Bury College in 1990. They kick-started their career with a show at the Corner Pin pub in Ramsbottom Manchester, and the rest is history. After releasing their debut album: Asleep In The Back in 2001, the band went on to win the Mercury Prize in 2008 with their hugely well-received fourth outing: The Seldom Seen Kid. The band has released seven studio albums, all of which (including their B-sides compilation: Dead in the Boot 2012), all reached the top 15 of the British album chart. Seven of their singles placed in the top 40 of the British singles chart. In 2008, Elbow won the Mercury Music Prize for their album: The Seldom Seen Kid, and in 2009 they won the Brit Award for Best British Group
One of Manchester's best for sure, Buzzcocks were formed in Bolton in 1976 around Howard Devoto and Pete Shelley. Buzzcocks were unchallenged pioneers of punk in Manchester, even putting on gigs and inviting the Sex Pistols to perform in the city for the first time. Self -motivated, their debut EP, Spiral Scratch, was self-produced on their own label and inspired countless other bands to join the DIY indie scene. Devoto left early on to form the band Magazine, but the band continued on and they achieved commercial success with singles like "Ever Fallen in Love" (With Someone You Shouldn't've) that fused pop craftsmanship with rapid-fire punk energy. These singles were collected on Singles Going Steady, a compilation that was described by critic Ned Raggett as a "punk masterpiece. Pete Shelley died on the 6thof December 2018.
Following the death of singer Ian Curtis in1980, the remaining members of Joy Division agreed to retire the name. Starting from scratch as New Order, the band added Gillian Gilbert on extra guitar and keyboards. As the 1980s progressed, New Order changed their sound as they became more interested in the electronica and production style that was taking hold of the music scene. Straight out of the box the produced the biggest selling 12" single of all time with the classic: "Blue Monday." Despite bassist Peter Hook leaving, the band returned in 2015 with a new album, Music Complete.
The band was formed in Oldham in 1983 by old school-friends Graham Lambert (guitar) and Stephen Holt (vocals). Inspiral Carpets really came to prominence when they recruited organist Clint Boon. Following his addition, the band was soon immersed into the burgeoning "Madchester" scene. Their songs, now sung by Tom Hingley, with the added quirkiness of Boon's psychedelic keyboards, gave them chart success in the early 90s with tracks like "This Is How It Feels" and "Move."
Formed in Salford in 1980, it took almost 10 years until the end of the decade that Shaun and Paul Ryder's band of trippy indie funksters signed to the legendary Factory Records. The band also featured their mate Bez as percussionist/dancer, and their unorthodox take on dance-rock fitted right in with the post-Acid House boom. The bands single: "Step On" hit the Top 5 in the UK in 1990, and even though the follow-up album Pills N' Thrills And Bellyaches was a success, the album pretty much bankrupted Factory. They split in 1993 but have reformed several times in subsequent decades.


text:
"So Good They Can't Ignore You"
When we look at people who are satisfied with their career, we will often see that they are passionate and love what they do. Successful people may be passionate, but does solely following your passion lead to success? As a young, inexperienced, and slightly restless university graduate who can no longer hide within the structures of student life, this question has been yearning for an answer.
One resource that has been a tremendous help for me is 'So Good They Can't Ignore You: Why Skills Trump Passion in the Quest for Work', written by Cal Newport (and available at McGill's CaPS Library!). The main thread that runs through the book is the importance of ability. It may be glazingly clear that no matter what you choose to do and why you do it, you have to be good at what you do. This simple advice differs from the popular career advice to "do what you love and the rest will follow". While it is crucially important to like your work, this piece of advice is incomplete.
Cal Newport tells a story of Steve Jobs that really rocked my boat. Etched in my memory is Steve Jobs' 2005 commencement speech to the graduating class of Stanford University. Jobs advised the newly graduates to "… find what you love" and "to follow your heart and intuition". In Newport's book, however, he tells the story of a younger Jobs and his passion for Zen Buddhism. In fact, Apple began as a way to make ends meet while pursuing his passion of Zen Buddhism. For Jobs, a new passion followed him once he becomes so good that the world could not ignore him.
The question is not whether you should follow your passion. In Newport's view, it doesn't matter what you choose. The only thing that matters is how you approach your work on a daily basis and your willingness to put in the work. Newport compares these two differing views in his book and refers to them as "the passion mindset" and "the craftsman mindset".
Craftsman Mindset
Passion Mindset
focus on work and improvement
focus on what work offers you ONLY
The craftsman mindset allows a person to solely focus on the quality of work. The only thing of concern is how to become good and valuable. All the secondary details of personality, personal appearance, connections, and equipment become what they are – secondary.
This concept of
following leading your passion through putting in the work is so simple that it's often missed.
I hope this short review has been helpful! Cal Newport explores so much more in his book. If you would like to read more, drop by the CaPS office and ask about the book!


text:
- bought wedding gown, dinner dress, wedding shoes
- design invitation card
- edit pre-wedding photos
- design wedding album
- create pictures montage/ slide show
To be continue....
Good luck gals :)ReplyDelete
thanks ;-) waiting for ur wedding too. :pReplyDelete
waiting for your wedding pic ^^
min-aris, thanks! Will share few pictures 1st at incoming post. ;-)ReplyDelete
I think everything will work out! ;) Congratz and my best wishes to you. :)ReplyDelete
Thanks, CH. ;-)ReplyDelete
hehe...soon!!! :p but only will upload few. ;-)ReplyDelete
wow wow...congrats to u!ReplyDelete
omigawd!! sound so exxciting!! heehee..
maybe this will help you with the flowers, you know those that you throw to your bridesmaids? I love that moment in weddings the most!
check the simple DIY Bouquet, purple roses cool!
Wishing you luck and joy on the process on your wedding~~ looking forward to see more of your wedding pictures~~ I'm taking mine this coming octoberReplyDelete
Vialentino, Thanks!
Nicolette, thanks for the link but i think i wont DIY the bouquet by own cos i'm will go back to hometown maybe 2 days before my wedding, so it is hard to get the flower there. :S So no choice, i've order from florist shop. It will more expensive.
Ciyou, thanks! ;-) I will post it once i complete my album. :pReplyDelete


text:
Picture this you a horny new female fuck buddy naked and having sex in public, Sounds out of this world doesn't it? I wonder what you would do if we said we can make this happen for you. I know what you would say, sign me up I want a bit of naked sex in public, Well my fellow doggers you can make it happen now! Simply register to UK dogging and in a matter of minutes start looking for a man/ woman to meet and enjoy naked sex in public with. Public sex can be anywhere in the pub toilets, your own back garden, cinemas anywhere and our horny sluts say the more riskier the public place the better. Myself and my wife have enjoyed a few naked encounters in public our latest one was the weekend just before christmas we were strolling along our local fields after a few shandys in the pub, the fresh air & drink got the better of us and we were soon tearing each others clothes and shagging right there and we were certainly not bothered about anyone catching us, Would of added to the thrill if they did.
There are tons of couples and singles right here on UK dogging who love to get naked and have sex in public and not only that you can meet up with our sexy flashers who are not afraid to walk round day or evening naked and flashing their bums, boobs and vaginas. UK dogging is where all the action happens and fantasies become a reality and we would love nothing more than to help you find the perfect couples/ sex partners for you, We have the biggest dogging database around and can guarantee you will be having naked sex in public in a matter of days. So what are you waiting for? Sign up to uk-dogging.co.uk for free now and get your naked public sex meet arranged.


text:
Dr Ian Smart
Ian is a leading consultant and entrepreneur. He analyses major investment decisions for business and government with a strong focus on evidence and data.
Ian is accredited to provide investment management services to the Victorian, Queensland, Australian, Western Australian and New Zealand Governments. He has facilitated more than 70 workshops and prepared more than 50 business cases for government investments over the last 10 years.
Ian has commercialised a number of ventures from the health sector. Cell Therapies was a joint venture with the Peter MacCallum Cancer Centre. Cell Therapies provides clinical trials in blood cell therapies for cancer patients. Through Cell Therapies the Peter MacCallum Cancer Centre turned a loss centre into a profit centre. Ian was also involved in the commercialisation of a prostate cancer drug, which was licensed to a pharmaceutical company.


text:
Somehow in my thrashing with SW2013, I accidentally maximized the Save As dialog box. It now appears full screen, so I can't look at the SW window behind it which is often useful when naming a new document. I can't drag the corner of the window to resize it, and there's no Windows button in the upper right corner except the X box to close the window.
How do I reduce the size of the dialog box?


text: A lady with an attitude. Sexiest stripper. The one to worship and obey to. I`m your secret lover! Ur Mistress& Goddess& Size Queen! Dirty talk, sexy voice, curvy body, soft pale skin, easy going, open minded and ready for kinky games cam2cam! I can be gentle or cruel- you choose! Join me in a world full of sexiness and orgasms!


text: The Savannah business directory currently lists 10.874 companies from a variety of industries. Find the right company quickly and easily in search Savannah! Simply enter the industry or the name in the search fields. The search finds the right company quickly and reliably. The acompio business directory still has 9.373.298 Profiles companies in United States of America. Search and find - your guide in Savannah.


text:
Sacred Heart of Jesus — Tough Love
It took me a while to appreciate the Sacred Heart of Jesus devotion. To tell the truth, as a convert, it all
It took me a while to appreciate the Sacred Heart of Jesus devotion. To tell the truth, as a convert, it all
Do you remember that creepy movie Invasion of the Body Snatchers? Some kind of alien bug invades earth and takes over people's
In 1914 Agnes Cuff, a flighty and unstable young woman with few prospects and little money found herself pregnant. The father didn't
there is "no medium in true philosophy between atheism and Catholicism… a perfectly consistent mind, under those circumstances in which it
I was talking with a man I'll call Henry today who lost his wife just before Christmas. She was only in her
Toppling the Pagan Idols Did the pagans really worship idols? If so, what was going on? To be precise, they
Have you noticed that nobody loves modern churches? Nobody. I mean NOBODY. Seriously. Have you ever met anyone who sees a church
An article in the Daily Telegraph ponders why the number of people attending parish churches in England continues to plummet. While
In re-reading C.S.Lewis' wonderful sermon The Weight of Glory he begins with comments on the nature of desire. Christianity is known by those outside
Margery Eagan, the spirituality columnist at Boston Globe's CRUX website poses a question about homosexuality in this column. Listening to the reading


text:
The Bahya grey marble washbasin has a small, discreet but stylish look. It can easily be placed in the corner of bathrooms and toilets. The authenticity of its gray marble, its welcoming curves and its stylish design are all a plus. The simple, but rich colour tone of the marble make it blend well with any kind of bathroom. The Bahya marble vanity uint is also available in black.
Internal dimensions (H 13 x W 29 x D 29 cm)
- plughole: 4.5 cm
- marble thickness: 3 cm
Tip: Be careful not to use abrasive cleaners
Use our carefully selected cleaning products to maintain the shine of your marble, lava stone or polished stone washbasin(s)
Routine maintenance
>> Buy the Cleaning product
Special maintenance
>> Buy the Renovator and Balm
Learn more : our furniture maintenance guide


text:
Do you need a lawyer in Illinois for a nursing home or medical related case? We will speak with you free of charge regarding your legal problem. Call 847 732 5986 right now for a free consultation with an Illinois attorney.
How will you find the right lawyer for your case?
SPEAK WITH AN ILLINOIS ATTORNEY RIGHT NOW. When you call 847 732 5986 you know who will answer the phone ... and you know his credentials and professional experience!
Michael Perillo is currently in his 36th year of active practice as an attorney. He is a former active duty U.S. Navy JAG Officer. Mr. Perillo has served as a Senior Assistant Public Defender in the Lake County, Illinois, Public Defender's Office and as a Senior Assistant State's Attorney in the Lake County, Illinois, State's Attorney's Office.
Mike Perillo has been engaged in the active practice of law for the past 35 years. His clients include people who have been seriously injured and others who were injured, abused or neglected in Illinois nursing homes. He is experienced in criminal defense and commercial and general civil litigation.
Mike Perillo is rated "A/V Preeminent," the highest rating available from the prestigious Martindale Hubbell Law Directory (www.martindale.com/ratings). He has also received a 10.0 rating on AVVO.
If you need a lawyer, you don't want to waste your time speaking with a non-attorney or an answering service. Speak directly with an Illinois attorney the very first time you call. Michael Perillo is available to take your call regarding your Illinois legal issue. Call 847 732 5986 now.
The telephone consultation is free of charge. You will only be responsible for attorney fees if you sign a contract for professional representation. Personal injury cases are accepted on a contingency basis, meaning the lawyer agrees to accept a fixed percentage of the recovery. If you win the case, the lawyer's fee comes out of the money awarded to you.
Mike Perillo accepts calls from throughout Illinois including, but not limited to, Lake County, Cook County, McHenry County, Waukegan, Chicago, Grayslake, Libertyville, North Chicago, Zion, Gurnee, Kankakee and Antioch.
Mr. Perillo is licensed to practice law in the states of Illinois and New York.
*All information displayed on this website is informational and shall not be deemed as legal advice. If you're currently dealing with an individual legal situation, you're invited to contact us through email or by phone. Until an attorney-client relationship has been established, we urge that you avoid sharing any confidential information. This is an attorney advertising website.
I'm busy working on my blog posts. Watch this space!
Please reload