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Kidney-specific WNK1 (KS-WNK1) is a variant of full-length WNK1. Previous studies have reported that KS-WNK1 is predominantly expressed in the distal convoluted tubule (DCT) where it regulates sodium-chloride cotransporter. The role of KS-WNK1 in other nephron segments is less clear. Here, we measured the expression of KS-WNK1 transcript in microdissected renal tubules and found that KS-WNK1 was most abundant in the DCT, followed by cortical thick ascending limb (cTAL), connecting tubule, and cortical collecting duct. A high K+ diet enhanced the expression of KS-WNK1 in the DCT and cTAL, selectively. It has been reported that a high-K diet suppresses Na+ reabsorption in TAL. To understand the role of KS-WNK1 in Na+ transport in cTAL and the regulation by dietary K+, we examined Na+ reabsorption using in vitro microperfusion in cTAL isolated from KS-WNK1-knockout mice and wild-type littermates fed either a control-K+ or high-K+ diet. Furosemide-sensitive Na+ reabsorption in cTAL was higher in KS-WNK1-knockout (KO) mice than in wild-type. A high-K+ diet inhibited Na+ reabsorption in cTAL from wild-type mice, but the inhibition was eliminated in KS-WNK1-KO mice. We further examined the role of KS-WNK1 using transgenic mice that overexpress KS-WNK1. Na+ reabsorption in cTAL was lower in transgenic than in wild-type mice. In whole animal clearance studies, a high-K+ diet increased daily urine volume and urinary Na+ and K+ excretion in wild-type mice, which was blunted in KS-WNK1-KO mice. Thus KS-WNK1 inhibits Na+ reabsorption in cTAL and mediates the inhibition of Na+ reabsorption in the segment by a high-K diet.
- kidney-specific WMK1
- potassium adaptation
- sodium
- thick ascending limb
with-no-lysine (wnk) kinases are a family of serine/threonine kinases comprised of four members (WNK1–4) in mammals. WNK1, the founding member of WNK family, was first identified by Xu et al. (27) in 2000 and found to be ubiquitously expressed in tissues. The main function of WNK kinases is to regulate renal handling of sodium (Na+) and potassium (K+) as is evident by findings that mutations in WNK1 and WNK4 genes cause a syndrome of hereditary hypertension and hyperkalemia, called pseudohypoaldosteronism type II (26). Many studies mostly in vitro, have suggested that WNK1 and 4 modulate the activities of various Na+ transporters and K+ channels in the distal nephron via catalytic and noncatalytic mechanisms leading to increased Na+ absorption and impaired K+ secretion (9, 10, 19, 29). Kidney-specific WNK1 (KS-WNK1) is a shorter alternatively initiated variant of full-length WNK1 (FL-WNK1) predominantly expressed in distal nephron segments (28). The transcription of KS-WNK1 is initiated by an alternative promoter in exon 4A, which resides in intron 4 between exons 4 and 5. Thus the transcript of KS-WNK1 is shorter, containing exon 4A and the remaining exon 5∼28 of full-length WNK1, and the encoded protein lacks the kinase domain encoded by exon 3 and 4 (4). Results of studies (14, 24) based on heterologous cells have suggested that KS-WNK1 functions as an antagonist of full-length WNK1 with respect to regulation of renal Na+ and K+ transporters.
To study the function of KS-WNK1, we have created KS-WNK1 knockout mice and transgenic mice that overexpress KS-WNK1 in the kidney (15, 16). In KS-WNK1 transgenic mice, the total and phosphorylated forms of sodium-chloride cotransporter (NCC) and sodium-potassium-chloride cotransporter (NKCC2) in renal cortex are reduced. These mice display renal Na+ wasting and lower blood pressure under normal Na+ diet (16). Conversely, KS-WNK1 knockout mice have increased expression of NCC and NKCC in renal cortex and hypertension when fed a high Na+ diet (16). Another group (8) has also generated KS-WNK1 knockout mice and reported similar findings of increased NCC abundance and mild expansion of the circulatory volume in the mice as evident by decreases in 24-h urine aldosterone. These two studies are consistent with KS-WNK1 being an inhibitor of NCC in the distal convoluted tubule (DCT). The role of KS-WNK1 on NKCC2, however, remains unsettled despite our studies on KS-WNK1-knockout (KO) and transgenic mice. This is in large part due to previous reports (4, 20) based on in situ hybridization that KS-WNK1 was not detected in thick ascending limb (TAL) and that there have not been direct functional studies of Na+ transport in this nephron segment.
Dietary K+ is an important modulator of Na+ reabsorption in the kidney. It has been reported that dietary K+ loading causes natriuresis in part from suppression of Na+ reabsorption in TAL (25). The underlying mechanism of this K+ adaptive response in TAL is unknown. We (14, 16) have previously demonstrated that a high K+ diet increases the expression of KS-WNK1 in whole kidney and transgenic mice that overexpress KS-WNK1 in the kidney have decreased NKCC2 expression. Here, we study the effect of KS-WNK1 on Na+ transport in TAL and its role in the regulation of sodium transport by dietary K+ using in vitro microperfusion.
We have created KS-WNK1 knockout mice (KS-WNK1-KO) by deleting exon 4A in 129/sv strain and transgenic mice overexpressing amino acid 1–253 of KS-WNK1 transgenic (KS-WNK1-Tg) in C57BL/6 strain (14, 16). All of the experimental procedures involving these animals were performed in accordance with relevant laws and institutional guidelines approved by the University of Texas Southwestern Medical Center at Dallas Institutional Animal Care and Use Committee.
Balance studies.
For plasma biochemical data in control and high K+ diet, 10 KS-WNK1-KO mice and control littermates (129/sv wild type) (4 female, 6 male, ∼4-mo old) were fed a control K+ (1% KCl, 1 g per 100 g diet) or a high K+ diet (10% KCl, in substitution for equal weight sucrose in the diet; Harlan Teklad, Madison, WI) with free access to tap water for 2 wk before the collection of blood samples. Both control and high K+ diets contain 0.74% NaCl (7.4 g NaCl/Kg food). Mice were anesthetized with isoflurane, and whole blood was drawn by retro-orbital bleeding into Na+-heparin-coated glass capillary tubes. Plasma was recovered immediately by centrifugation. Plasma electrolytes were analyzed using STAT-CCC analyzer (Nova Biomedical, Waltham, MA). Plasma creatinine level was measured by capillary electrophoresis (P/ACE MDQ; Beckman Coulter, Brea, CA) as described previously (16). For balance studies, 10 KS-WNK1-KO mice and control littermates (129/sv wild type; 4 female, 6 male, ∼4 mo old) were fed the indicated diet and water ad libitum and placed in metabolic cages (Hatteras Instruments, Cary, NC) for collection of urine. The concentrations of Na+ and K+ in urine were measured using a microflame-emission photometer (Jenway, Essex, UK). The osmolality of plasma and urine was measured using an osmometer (model 3D3; Advanced Instruments, Norwood, MA). The osmolar-free water clearance (CH2O) was calculated by the equation: A positive value indicates that the kidneys excrete solute-free water and dilute urine. Conversely, a negative value reflects free water reabsorption and the kidneys excrete concentrated urine.
Preparation of mice for microperfusion and microdissection studies.
These experiments were performed in KS-WNK1-KO and KS-WNK1-Tg mice at 8–10 wk of age and age- and gender-matched wild-type littermates (129/sv strain for KS-WNK1-KO and C57BL/6 strain for KS-WNK1-Tg). The mice were raised in a 12-h day and night cycle and fed a control K+ (1% KCl) or a high K+ diet (10% KCl) and tap water ad libitum for 2 wk before death.
In vitro microperfusion, sodium flux, and transepithelial potential difference.
After the mouse was killed, the kidney was removed quickly, sliced in thin coronal sections and placed in Hanks' solution containing the following (in mM): 137 NaCl, 5 KCl, 0.8 MgSO4, 0.33 Na2HPO4, 0.44 KH2PO4, 1 MgCl2 10 Tris(hydroxymethyl)amino methane hydrochloride, 0.25 CaCl2, 2 glutamine, and 2 l-lactate at 4°C. The cortical thick ascending limb (cTAL) of Henle's loop was then dissected free hand without collagenase and transferred to a 1-ml temperature-controlled bathing chamber. Tubules were perfused in vitro as previously described (22).
Isolated cTALs were perfused at a rate of ∼5 nl/min. The perfusate contained the following (in mM): 115 NaCl, 25 NaHCO3, 2.3 Na2HPO4, 10 Na acetate, 1.8 CaCl2, 1 MgSO4, 5 KCl, 8.3 glucose, and 5 alanine and had an osmolality equal to that of the bathing solution that contained 6 gm/dl of albumin. There were at least 3 measurements of the perfusion and the collected tubular fluid in each experiment. Na+ transport (JNa) was calculated using the equation: JNa (pmol·min−1·mm−1) = ([Na]perfusate − [Na]collected)(V̇L)/L, where V̇L is collection rate (∼5 nl/min) and L is the tubular length (0.4–0.8 mm). The transepithelial potential difference was determined using the perfusion pipette as a bridge into the tubular lumen and referenced to the bathing solution using a Keithley 6517A programmable electrometer (Cleveland, OH). Furosemide used to inhibit NKCC2 was purchased from Sigma-Aldrich (St. Louis, MO). Na+ concentrations of perfusate and collected drops were measured using a Na+-selective electrode (Sodium Ionophore II-Cocktail A; Fluka) as previously described (2, 17).
Microdissection and RT-PCR.
Slices of kidney were placed into prewarmed collagenase type I (Worthington, Lakewood, NJ; 1.5 mg/ml dissolved in DMEM/F12) solution in 15-ml test tube, which was then shaken vigorously on a titer plate shaker at 37°C for 10–15 min. After digestion, the individual nephron segments were dissected in 4°C Hanks' solution and transferred by adhering the tubules to small glass beads (0.5-mm diameter; Thomas Scientific, Swedesboro, NJ) and then transferring the beads to 1.5 ml tubes containing 0.6 ml RNase inhibitor-containing lysis buffer. Total RNA was immediately extracted using Quick-RNA MicroPrep kit (Zymo Research, Irvine, CA). Reverse transcription was performed using TaqMan reverse transcription reagents (Applied Biosystems, Carlsbad, CA). Quantitative real-time PCR was carried out on MyiQ single color RT-PCR detection system (Bio-Rad, Hercules, CA). We verified that no amplification was produced when reverse transcription was omitted in each sample. Sequences of primers for RT-PCR analysis were provided in Table 1. Relative mRNA levels of target proteins were standardized with an internal control (GAPDH). For comparison between KS-WNK1 and FL-WNK1, the efficiencies of KS-WNK1 and FL-WNK1 primers in RT-PCR assay were calculated from the results of three serial (4-, 16-, 64-fold) dilutions of cDNA, which encompass our working dilution (∼10-fold dilution) (21). The slope of threshold cycles obtained from KS-WNK1 or FL-WNK1 PCR reactions of serially diluted sample tubular cDNA was used to calculate the corresponding efficiencies (E) according to the equation: . Each sample was assayed in triplicate.
Statistical analysis.
All results are expressed as means ± SE. Statistical comparisons between two groups of data were made using two-tailed unpaired Student's t-test. Multiple comparisons were determined using one-way ANOVA followed by Tukey's multiple comparison tests. P value <0.05 was considered to be statistically significant.
Characterization of WNK1 isoform mRNA expression in renal tubules in control and high potassium diet.
The relative expression of WNK1 isoforms including FL-WNK1 and KS-WNK1 in individual renal tubules has not been studied. We measured relative mRNA abundance of KS-WNK1 and FL-WNK1 in renal tubules including the medullary thick ascending limb (mTAL) and cTAL, DCT, connecting tubule (CNT), and cortical collecting duct (CCD) using quantitative RT-PCR. The purity of these samples was confirmed by measuring the mRNA abundance of nephron segment-specific markers (NKCC2 for TAL, NCC for DCT, and aquaporin-2 for CNT and CCD; Fig. 1). To allow comparison of abundance of KS-WNK1 and FL-WNK1, we first determined efficiencies of PCR primers used in KS-WNK1 and FL-WNK1 RT-PCR assays (position of primers shown in Fig. 2A). The calculated efficiencies of KS-WNK1 and FL-WNK1 RT-PCR assays were 1.94 and 1.92, respectively (Fig. 2B), supporting the legitimacy of direct comparison between two RT-PCR assays (21).
Under control K+ (1% KCl) diet, the expression of KS-WNK1 was most abundant in the DCT but was also present in the cTAL, CNT, and CCD (Fig. 2C, left). The KS-WNK1 transcript was almost undetectable in mTAL and proximal tubule (not shown for proximal tubule). As for FL-WNK1, its mRNA expression was more evenly distributed along the nephron in lower abundance compared with KS-WNK1 (Fig. 2C, right). The ratio of KS-WNK1 to FL-WNK1 mRNA is ∼18-fold in the DCT but closer to 1 in other segments (0.85, 2, and 0.5 for cTAL, CNT, and CCD, respectively).
To test the effect of a high K+ (10% KCl) diet on KS-WNK1 and FL-WNK1 mRNA expression, mice were fed either a control or high K+ diet for 2 wk before isolation of renal tubules. A high K+ diet increased KS-WNK1 mRNA expression in cTAL and DCT (90 and 60% increase, respectively) but not in other segments (Fig. 3A). In contrast, a high K+ diet had no effect on FL-WNK1 mRNA (Fig. 3B).
KS-WNK1 mediates high potassium diet-induced suppression of sodium reabsorption in the cTAL.
We (16) have previously shown that overexpression of KS-WNK1 decreases protein abundance of total and phosphorylated NKCC2 in the TAL and knockout of KS-WNK1 has opposite effects. Here we studied the physiological relevance of these findings by measuring Na+ reabsorption in isolated cTAL using in vitro microperfusion. In cTALs isolated from wild-type mice, Na+ reabsorption occurred at a rate 175 ± 12 pmol·min−1·mm−1 (Fig. 4A). This reabsorbed flux agrees with previous reports in cTAL (18). Addition of furosemide (100 μM) to perfusate caused ∼90% reduction of Na+ reabsorption in cTAL (Fig. 4C), indicating that it is mediated by NKCC2. A high K+ diet suppressed Na+ reabsorption in cTAL by ∼24% (Fig. 4A). In KS-WNK1-KO mice, the baseline Na+ reabsorption in the cTAL under control K+ diet was ∼20% higher than control (Fig. 4A). High K+ diet did not significantly suppress Na+ reabsorption in the cTAL of KS-WNK1-KO mice (Fig. 4A, right). Na+ reabsorption in TAL via NKCC2 generates a lumen-positive transepithelial potential difference (PDTE ). A similar pattern was observed in measurements of PDTE (Fig. 4B).
To further support the functional role of KS-WNK1 in cTAL, we measured Na+ flux in cTAL of transgenic mice that overexpress KS-WNK1 in the kidney. Na+ reabsorption in cTAL was ∼33% lower in KS-WNK1-Tg mice than in control littermates under control K+ diets (Fig. 5A). A high K+ diet did not cause further reduction of Na+ reabsorption in cTAL from KS-WNK1-Tg mice. Similar changes were observed in PDTE measurements (Fig. 5B). These functional results, together with our (16) previous biochemical and immunological analysis of NKCC2 in KS-WNK1-KO and KS-WNK1-Tg mice, indicate that KS-WNK1 inhibits NKCC2 in cTAL and that KS-WNK1 mediates the inhibition of Na+ reabsorption in cTAL by a high K+ diet.
KS-WNK1 knockout blunts the high potassium diet-induced natriuresis.
A high K+ diet causes natriuresis by affecting Na+ reabsorption at multiple nephron segments (1, 25). To understand the contribution of KS-WNK1-mediated inhibition of NKCC2 to the overall natriuretic response in renal K+ adaptation, we performed whole animal clearance studies. Mice were fed a control K+ diet for 4 days and then switched to a high K+ diet for 7 days. Daily urine volume and urinary Na+ and K+ excretion during control K+ diet were not different between KS-WNK1-KO and wild-type mice. A change to a high K+ diet resulted in an increase in urine volume in both wild-type and KS-WNK1-KO mice starting from the first day of high K+ diet (Fig. 6A). The high K+-induced diuretic response, however, was blunted in KS-WNK1-KO mice relative to wild-type littermates.
We next examined the natriuretic response to a high K+ diet. Unexpectedly, urinary Na+ excretion in both wild-type and KS-WNK1-KO mice decreased on the first day of high K+ diet (Fig. 6B). This initial drop in urinary Na+ excretion likely reflected decreased intake of the high K+ diet due to palatability of the diet (see Table 2). A similar observation has been made in mice by others (3, 20). After the first day, urinary Na+ excretion increased in both wild-type and KS-WNK1-KO mice reflecting natriuretic response to a high K+ diet. Compared with wild-type, the natriuretic response in KS-WNK1 mice was somewhat blunted on the second and third day of a high K+ diet (Fig. 6B, inset). It should be mentioned that the increase in urinary Na+ excretion during high K+ diet may reflect alterations in the extra-renal Na+ excretion as well as renal Na+ excretion. In the control K+ diet, urinary Na+ excretion was ∼59% of intake (261 μmol over 443 μmol), indicating that ∼41% of ingested Na+ was excreted extra-renally, presumably mostly by the intestine. In the high K+ diet, urinary Na+ excretion slightly exceeded the amount that was ingested [362 μmol excreted vs. 342 μmol Na+ ingested (0.74% × 2.7 g diet)]. We speculate that increased aldosterone secretion in high K+ diet increases intestinal Na+ absorption, which together with the renal natriuretic effect, results in urinary Na+ excretion exceeding that is ingested.
As expected, urinary K+ excretion increased on a high K+ diet (Fig. 6C). Likely because of the decrease in intake of the high K+ diet (and increased intestinal K+ secretion as well; see below), daily urinary K+ excretion increased only by ∼7-fold rather than the expected 10-fold increase. As for the natriuretic response, the increase in K+ excretion was less in KS-WNK1-KO mice relative to wild-type mice on the second and third day.
The above results support the hypothesis that KS-WNK1 contributes to high K+ diet-mediated diuresis and natriuresis. Serum K+ levels, however, were not different between KS-WNK1-KO and wild-type mice, even on a high K+ diet (Table 2). Thus other mechanisms must fully compensate for the transient K+ excretion impairment caused by loss of KS-WNK1. Of note, the difference in urinary K+ excretion between wild-type and KS-WNK1-KO mice on day 2 and 3 of high K+ diet apparently far exceeds that can be potentially buffered by the intracellular space. We did not detect significant differences in the daily food intake between wild-type and-KO mice (not shown), which was supported by the finding that urinary osmolar excretion was not different (Table 2). Thus increased intestinal K+ excretion may also contribute to the compensatory responses in-KO mice. An increase in aldosterone secretion, as mentioned above, may be a compensatory mechanism for increased intestinal K+ excretion in high K+ diet. However, we have no explanations why increases in the intestinal K+ excretion would be greater in KS-WNK1-KO than in wild-type mice.
Interestingly, urine osmolality is significantly higher in KS-WNK1-KO than in wild-type in control K+ as well as high K+ diets (Table 2). Moreover, the increase of urine volume on the seventh day of a high K+ diet was much higher than the increase of urine osmolar excretion (Table 2; ∼6-fold vs. ∼3-fold). As the total daily urine osmolar excretion was not different between the two groups on either a control or high K+ diet, these results suggest that potassium-induced diuresis has more than an effect on osmotic diuresis, and may be in part due to a decrease in free water reabsorption (−CH2O; −5.9 ± 0.4 μl/min [control K+] vs. −4.7 ± 0.7 μl/min [high K+]; P < 0.05; Table 2). Interestingly, free water reabsorption was higher in KS-WNK1-KO mice than in wild-type mice. These results are consistent with the idea that KS-WNK1 inhibits NKCC2, and increased NKCC2 activity in WNK1-KO mice enhances urinary concentration ability.
Using quantitative measurement of mRNA and in vitro microperfusion, we show in this study that KS-WNK1 is expressed in cTAL and plays an important role in inhibition of NKCC2-mediated Na+ reabsorption in the segment. Moreover, KS-WNK1 mediates the high K+ diet-induced inhibition of Na+ reabsorption in cTAL and is important for urinary concentration.
Since the first discovery of KS-WNK1 in 2002 (28), two studies (4, 20) using in situ hybridization of KS-WNK1 mRNA have shown that the expression of KS-WNK1 is exclusively in the renal cortex and that the DCT has strikingly high KS-WNK1 expression. Using a more quantitative and sensitive method, quantitative RT-PCR, we show that while KS-WNK1 transcript is most abundant in DCT, and it is also present in cTAL, CNT, and CCD; the abundance in these segments relative to DCT are 30, 17, and 8%, respectively. We confirm that KS-WNK1 is barely detected in mTAL and the proximal tubule. The physiological importance of KS-WNK1 in Na+ reabsorption in cTAL is demonstrated by microperfusion studies.
Transepithelial Na+ reabsorption in cTAL is mediated in large part by the transcellular route, which requires Na+ entry across the apical membrane predominantly through NKCC2 and exit via the basolateral Na+-K+-ATPase. NKCC2-mediated Na+ reabsorption generates a lumen-positive PDTE, partly due to apical recycling of K+. The Na+/H+ exchanger also contributes to the apical Na+ entry, but the role is relatively minor and it does not generate lumen-positive PDTE (23). Several pieces of evidence indicate that the effect of KS-WNK1 on Na+ reabsorption in cTAL is at least in large part through inhibition of NKCC2. First, it is inhibited by the NKCC2 inhibitor furosemide. Second, the effect on Na+ flux parallels that on lumen-positive PDTE. Third, and perhaps more importantly, we (16) have previously shown that KS-WNK1 negatively regulates NKCC2 abundance by Western blot analysis and immunofluorescent staining. The additional effect of KS-WNK1 on other transporters in cTAL, nonetheless, is possible. In this study, we did not directly examine the mechanism by which KS-WNK1 inhibits NKCC2. However, results of studies based on Xenopus oocytes and cultured cells (14, 19, 24) suggest that KS-WNK1 probably antagonizes the activation of NKCC2 by FL-WNK1.
The other significant finding of our study is that KS-WNK1 appears to mediate high K+-induced inhibition of Na+ reabsorption in cTAL. It is known that a high K+ diet inhibits Na+ reabsorption in the proximal tubule and TAL (1, 11, 25). We (14) have previously shown that a high K+ diet upregulate KS-WNK1 mRNA in the whole kidney lysate. Here, we further show that high K+ diet upregulates KS-WNK1 mRNA in cTAL as well as DCT but not in other segments. In microperfused cTAL, a high K+ diet inhibits Na+ reabsorption, and the inhibition is obliterated in KS-WNK1-KO mice. Thus KS-WNK1 is important for inhibition of Na+ reabsorption in cTAL by a high K+ diet. One important physiological role of a high K+-diet induced natriuresis and diuresis is to maximize renal K+ excretion during high K+ intake. The renal K+ excretion occurs predominantly by secretion via K+ secretory channels in CNT and early CCD that requires lumen-negative PDTE generated by ENaC-mediated Na+ reabsorption. Several factors limit K+ secretion via this mechanism. First, continuing K+ secretion on top of water reabsorption accompanying Na+ reabsorption in the CNT and CCD raises the luminal K+ concentration to an extent that it favors reabsorption rather secretion in late CCD and the downstream outer medullary collecting duct. Second, Na+ reabsorption in the more proximal segments reduces delivery to distal segments. Inhibition of Na+ reabsorption in the proximal tubule and TAL by high a K+ diet would increase Na+ and fluid delivery to the CNT and CCD to overcome the above limiting factors. Moreover, increased flow to distal nephron segments also stimulates flow-activated maxi-K channels adding to K+ secretion. To our surprise, we found that urinary K+ excretion was only transiently decreased in KS-WNK1-KO mice in the second and third days of a high K+ diet. Serum K+ levels and urinary K+ excretion at day 4 of a high K+ diet and onward were not different between KO and wild-type mice. Apparently, other factors, such as inhibition of Na+ reabsorption in the proximal tubule by high K+ diets, compensate for the deficiency caused by loss of KS-WNK1.
It is well known that Na+ reabsorption by medullary TAL is critical for establishment of hypertonic medulla essential for the urinary concentration mechanism (6). Water restriction increases NKCC2 abundance in TAL and vasopressin stimulates trafficking and insertion of NKCC2 to the apical membrane (5, 7). Bartter's mutations in NKCC2 and pharmacological inhibition of NKCC2 impair the urinary concentration mechanism (12, 13). The role of cTAL in the urinary concentration is less clear. Our results demonstrate that KS-WNK1 plays a role in the urinary concentration mechanism. KS-WNK1-KO mice have a higher baseline urinary osmolality and free water reabsorption. We speculate that this is due to increased NKCC2 activity in the cortical TAL of KS-WNK1-KO mice. The role of NKCC2 in medullary TAL is unlikely because KS-WNK1 is not expressed there. The possibilities that KS-WNK1 regulates aquaporin-2 and urea transporters also exist and require further investigation.
This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases Grants DK-41612, DK-59530, and DK-079328. C. J. Cheng is supported by a scholarship grant from the Ministry of Defense, Taiwan.
No conflicts of interest, financial or otherwise, are declared by the author(s).
Author contributions: C.-J.C., M.B., and C.-L.H. conception and design of research; C.-J.C. and T.T. performed experiments; C.-J.C., T.T., and C.-L.H. analyzed data; C.-J.C., T.T., M.B., and C.-L.H. interpreted results of experiments; C.-J.C. prepared figures; C.-J.C. drafted manuscript; M.B. and C.-L.H. edited and revised manuscript; C.-L.H. approved final version of manuscript.
We thank Jessica Lucas for measuring creatinine concentrations of plasma and urine samples. This work was performed by C. J. Cheng in partial fulfillment of the requirements of the Ph.D. degree at the University of Texas Southwestern Medical Center.
- Copyright © 2012 the American Physiological Society


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German Shepherd rescue remembers fallen officer
Officer Rod Bradway was a true public servant, not only working to serve and protect people, but also animals.
He and his wife collected a trailer full of animal supplies to take to Moore, Oklahoma after the tornadoes hit there in May. This is just one of many examples.
That's why the Indy City Market expanded its services Wednesday to include an animal supply drive for Indianapolis Animal Care and Control in honor of Officer Bradway. People gave animal toys, treats, food and cleaning supplies - even cash donations - to help the shelter keep up with the more than 700 animals they regularly care for.
Most recently, Officer Bradway gave the majority of his free time to one animal rescue in particular, providing a new home to animals who needed it most. The owner of German Shepherd Indy Rescue is still in shock at such a big loss to the local animal community.
To say Amber Marks loves her job is an understatement. She gave up a career in corporate America to sling it out in the trenches with a lively bunch that make up German Shepherd Rescue Indy. These German Shepherds never complain. They never criticize. They only love and love to be loved.
Perhaps that's what Officer Bradway saw when he showed up one day two months ago at the request of a mutual friend to help Amber prepare her new, larger shelter.
"The first time he was here, he was here over 10 hours, hard labor. It was almost like he shared my vision and just did whatever it took to get it to fruition," said Marks.
The public servant didn't discriminate between man and animal when it came to serving and protecting.
"He secured all of the fencing up above the kennel walls. So, he helped secure that. He was pivotal in making everything come together so I could bring the shepherds here and save more," said Marks.
Nineteen dogs, 19 different stories - Red was found abandoned on the highway.
"I named him Bradway because he deserves a chance. He's a beautiful man and he's going to get a great home, I'm going to make sure of it and this is to honor Officer Bradway," said Marks.
Just as Officer Bradway gave another dog a chance.
"He took in a dog, him and his wife, who suffered neglect and cruelty and brought the dog back to health and just adopted it," said Marks.
Suggested Links
The Bradways just adopted Tucker last week. It's a final act in a heart wrenching story that closes with at least one happy ending.
"Someone that has a huge heart. It's like they're born to give," said Marks.

subset: commoncrawl


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What To Expect
Experience some of the most spectacular views in the world. The tour begins with a scenic drive along the Atlantic coast passing through the fishing village of Hout Bay. Here you will have the option of taking a short boat trip to Duiker Island to view the Cape Fur seals (own account).
The tour then heads along Chapman's Peak*. Drive on to Cape Point and the Cape of Good Hope Nature Reserve. Return to Cape Town by way of the Simon's Town naval base where we visit the penguin colony at Boulders Beach (optional, for own account). Travel via Fish Hoek fishing village and Muizenberg to Kirstenbosch Botanical Gardens for a short walking tour.
Meals and drinks
Tips and gratuities
Personal expenses
Pick-up from other hotels may be requested at the time of booking.
Departure time and duration vary depending on hotel location.
This tour is non-refundable if cancelled 14 days or less prior to the chosen tour date.
All our card payments are protected by thawte to give you peace of mind.


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As the Supreme Court moves towards a decision on President Obama's health care reform law, there are several possible directions in which it may go -- and none of which may even be carved in stone when a "final" outcome has been determined. In any regard, though, it is essential to have a firm plan in place going forward.
The court has several additional issues to decide upon, such as the constitutionality of potential IRS penalties for not purchasing one's own health insurance, as well as the coerciveness of Medicaid expansion.
Many Americans have been led to believe that this health care reform was designed to go after the supposedly "big and bad" insurance companies for making too much profit via their health insurance policies.
While it may be difficult to predict exactly which way the tide will turn, there are a number of factors that lead us to believe that the mandate requiring the purchase of health insurance will not stick -- possibly leading to the law being struck down altogether.
Square One
By using pre-existing condition policy rejections, along with stating that policy maximums for catastrophic health care claims are currently too low, insurance companies are surely being made out to be the "fall guys" in the overall framework of this reform.
Yet, the reality is that a large percentage of big employer sponsored health insurance plans are actually self-funded, meaning that they aren't even using insurance companies' coffers for covering their potential risk.
In addition, regarding benefit maximums on individual health insurance plans, statistically only about 1 percent of claims that are filed will typically even come close to exceeding the stated policy max - so the raising of these figures is really a non-event.
This, coupled with the fact that the health care law was hastily moved through Congress, has essentially led to a great deal of confusion - not just for the American people, but also for those who are directly involved in its passing.
One big reason for this includes the fact that the Justice Department's own attorneys were not even involved in crafting the law, nor did the proposed reform go through the Judiciary Committee. So, with neither of these government resources even being used in its creation, chances are good that it very well may be deemed as unconstitutional.
Possible outcomes
There are a number of ways that the Supreme Court could go on this topic moving forward. First, the law could be upheld altogether -- albeit such a decision could potentially unleash a series of legal, political, and legislative forces.
Another option would be simply that more of the law's provisions would be invalidated. In this case, it would seem that lawmakers would be under less pressure to continue making adjustments if the insurance pre-existing conditions and related changes go away -- along with the mandate. In this case, the insurance industry would still closely resemble that of today -- although it is likely that the quality of coverage would be negatively affected, primarily for those who are old and / or in poor health.
In narrowing down the possible outcomes of the Supreme Court decision, our consensus is that the Court will strike down the individual mandate due to its unconstitutional nature. Should that be the case, it could be months or even years before any of the proposed issues are resolved, putting the health insurance industry -- and those who utilize its services -- firmly back to square one. The question still remains, though, what can you do in the mean time?
The Bottom Line
Regardless of which direction the Supreme Court takes, it is highly likely that there will still be lawsuits, state actions, and even additional battles within Congress in both the short- and long-term. To learn more about how the Supreme Court health care ruling will impact your business visit www.supremecourthealthcareruling.com.


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William Wissing, Dutch ca.1656 - 1687Though from Amsterdam, Wissing's early training took place under Willem Doudyns, a historical painter of The Hague. From there it is believed that he went to France where he acquired a passion for metallic flowers, huge weeds and fancy accessories. It is presumed he entered Sir Peter Lely's studio upon arriving in England in 1676. Wissing came to be considered the most important of Lely's later pupils, and grew into fashion after the death of his master. The artist found obvious favor with the nobility, to the point of painting the royal family members. Apparently his good manners and gentleness recommended him well. Descamps, in his "Life of Wissing
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Siblings pass test of time
ATHLETICS Sisters Maayan and Lia Radus were among the young Jewish athletes to improve their best performances in the Watford Open Graded Meeting series.
Maayan Radus, 10, recorded a 1,500m best of 5:33.88 to place third in the UK under-11 rankings, while Lia, 12, cut her respective 800m and 1,500m times to 2:31.10 and 5:10.98.
Their Shaftesbury Barnet clubmates Jessica Hurley and Judah Yawitch also set 800m PBs at Watford. Hurley, 14, recorded 2:21.36 while Yawitch, 11, improved his best to 2:27.56. He also sealed victory in the Eastern Young Athletes League at Stevenage with a new 1,500m best of 5:02.9. Colette Hurley, mother of Jessica, moved to seventh in the UK V50 400m rankings by recording 69.24 at Watford.
Sophie Dowson, 15, took victory in the Saucony Hampshire Grand Prix in Basingstoke, with a pole vault clearance of 3.20m. Allana Edwards, 14, recorded 50.27inwinningthe300mhurdlesevent in the Trafford Grand Prix at Stretford.


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Finding a Good Topic for an English Essay
Aug 23, 2012 - Posted to
Essay Writing
One of the most difficult parts of writing an
English essay
is finding a good topic. Sometimes, your tutor can give the essay task to you with assigned topic. Other times, you are left to your own devices. This difficult situation can be made a lot easier if you know what to do.
rainstorming is a good way of choosing an English essay topic. You may choose to brainstorm with your classmates, course mates, colleagues or even your professor. Your purpose here is for you all to discuss particular areas that you find interesting. When you brainstorm, you are looking to pick points in your favor. When you hear a colleague say something interesting in your brainstorming session, quickly write it down. Ask your colleague or friend to explain what he means. Do this repeatedly and you may already have an English essay topic.
ist all the topics that arouse your interests in a jotter or piece of paper. Sieve this list to separate the more interesting topics from the less interesting ones. Put your list in a safe place for later use.
he reason you are in school is to learn. It then follows that you must have acquired some knowledge down the line. On a separate piece of paper write down all the topics that you are knowledgeable about. You might even add that you know "the earth is round". This is accepted, provided you have proof to back it up. You will find writing your English essay easier if you concentrate on subjects that you have information on.
ring out your 3 lists. Think about the topics you already have listed. Ask yourself which of these topics you can honestly do justice to. Can you get closely involved in any topic without being lost in the ideas? It is absolutely important that you research any topic you decide to write about.
ake your topic interesting. A boring topic will not keep the attention of your readers for long. If it is possible, choose a topic that has not be covered during your classes before. If you find this hard to do, approach your topic from a different perspective. If possible use the
DRAPES method
to capture your readers and keep them coming for more.
Now that you are writing or may have written your English essay, consider it as a first draft. There is no such thing as a completed essay. All essays are simply works in progress.
Proofread and edit your essay
several times. When you are done with this, pass it on to a second party. You will find a second person may notice errors you over looked the first time.
Read your essay aloud to yourself. Does it sound interesting of flat? Read it again to another person. What does this new person have to say about your English essay? Is your essay on focus? Is it organized? Does it have supporting evidence? What about the length and citations are they in order? Take note of these questions and you may be close to writing a captivating essay.
On a final note, I'd advice writing your first draft without worrying about grammar and spelling. For the second draft you should eliminate these things. Subsequent drafts should focus on other aspects of your essay.
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The Walking Dead Season 8 out October
The Walking Dead Season 8 will be released date confirmed via a poster, to be released October 22.
The production of Season 8 has been smooth, after the accidental death of stuntman John Bernecker last week. Bernecker suffered a fatal head injury after he fell 20 feet onto a concrete floor while rehearsing a fight scene.
We are all praying for him, his family, friends and colleagues. https://t.co/03Kj5JLwun— Gale Anne Hurd (@GunnerGale) July 13, 2017


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Adj Handle, M10, Ext, SS, 0.99, 2.93, MD, NG
KIPP| Zoro #: G0225303| Mfr #: K0270.2101X25
Zoro #: G0225303Mfr. Part #: K0270.2101X25
$13.89 /ea
Orders over $50.00 ship free. DetailsStandard ground shipping
Technical Specifications
|Zoro #: G0225303 Mfr #: K0270.2101X25
|Finish: matte
|Style: Novo Grip
|Color: Black
|Knob Type: Modern Design
|Material: Plastic
|Item: Adjustable handle
|Thread Size: M10
|Type: External Thread, Stainless Steel
|Height: 2.62"
|Screw Length: 0.99"
|Overall Length: 2.93"
|Height (In.): 2.62
|Country of Origin (subject to change): Germany
Product Description
Adjustable Handle, External Thread, Stainless Steel, Screw Length 0.99 In, Overall Length 2.93, Thread Size M10 In, Height 2.62 In, Knob Type Modern Design, Novo Grip Style, Plastic, Matte


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Like/Dislike Stamps
- Self inking stamps that last up to approximately 5,000 uses
- Great way of showing approval/disapproval without actually speaking
- Pack contains 1 x Like stamp and 1 x Dislike stamp
- Dimension of each stamp: H:80mm/W:65mm/D:30mm
- Stamps message in clear blue ink
More product info
Now with the Like/Dislike Stamp you can authoritatively show your pleasure or displeasure at pretty much anything you can stamp on. For thousands of years, we've been giving the thumbs up or the thumbs down gesture - it even dates back to the ancient Romans when bloodthirsty spectators used to instruct gladiators as to whether they should spare their opponents or not. Now in the 21st century, social networking sites such as Facebook offer it as a source of opinion.
With the ink in each stamp lasting for up to 5000 assertions, you're free to show your feeling on things of your choice to the world! Organize your never-ending medley of takeaway menus with a definitive answer on whether or not you'd order from them again, cast your decision on that dreaded school report due later this month or simply show the world your feeling on anything, at any time! There is nothing like giving the silent treatment when annoyed; give the rubber ear by literally rubber stamping!
The novelty and fun of these stamps won't run out until you stop liking and disliking things! Please Note: This product is not associated with Facebook. Any trademarks that may be depicted are the property of their respective owners. No connection between such owners is implied.


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BF3 Main Theme Remix
Credits & Info
- Uploaded
- Sep 5, 2012 | 10:23 PM EDT
- File Info
- Song
- 3 min 11 sec
- Score
- 4.37 / 5.00
Related Content
You might also enjoy:
Licensing Terms
Please contact me if you would like to use this in a project. We can discuss the details.
- Score:
- Rated 4.37 / 5 stars
- Plays & Downloads:
- 2,843 Plays | 382 Downloads
- Share Links:
- Genres:
- Electronic - Video Game
- None
Author Comments
Rated 4 / 5 stars
ah, excellent song! gave me the chills because i'm such a big battlefield fan.
i was getting really excited and having known your style from listening to your songs from the past, i was expecting something a bit more around 0:59, something heavy with big bass. i wasn't looking for a whole different sound, but something i could have used in my youtube videos.
imagine LOTS AND LOTS of action and explosions, i need something heavy and heart racing with alot of bass and bangs and stuff, you know what i'm saying?
although, this is definitely the best remix on NG. excellent job
Rated 4.5 / 5 stars
All that FM. It's beautiful. :3
Rated 4 / 5 stars
Pity there wasn't a drop, that would've ensured a score of five from me. However, it's still a good song nonetheless. Not really much else I can say.
4 4 Fav Download
Total Win


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Camano Island Creativity
Northwest by the numbers
Wednesday, March 5, 2014
One gets the impression Karla Matzke can do anything: she paints, sculpts in stone and stainless steel, wrote the salient text of the recently published book 100 Artists of the Northwest, operates her gallery in a sculpture park she carved out of the woods on Camano Island and, not too long ago, curated and hung a stunning show of the works of 25 artists she represents (including herself).
The exhibit—which features those artists, who are also included in the book—opened last week with a reception and potluck, and will be on display through April 13 at the eponymous Matzke Gallery, located on Camano Island. To reach it, drive south on Elger Bay Road, which becomes S. Camano Drive. One mile past the gas station and deli—just past the spaceship—turn left into Blanche Way and follow the signs.
Her well-lit show—yes, Matzke hung the lights, too—gives plenty of space to some of the best artists in the Northwest. There aren't any slouches in the exhibit and if I don't mention some, it's because of lack of space.
Bellingham is represented by five luscious oils by David Ridgway, who transforms views of homes and barns in the woods into interlocking colored shapes and playful patterns. His small painting, "Yellow Barn," is so welcoming you'll want to fall right into it. Also from his hometown is Shirley Erickson, working wonders with steel and bronze linked to fused and cast glass, resembling found icebergs.
Peregrine O'Gormley of La Conner presents his strong, massive animal and human forms full of gravitas. Among them, a fierce hawk carved from red cedar ferociously holds its ground under sheltering wings (enigmatically entitled "Minus 25%"). They harmonize beautifully with the nice forest studies by Kathleen Faulkner (Anacortes) in pastel and charcoal.
James Madison has a strong presence in wood and metal sculpture. He is a Coast Salish Tlingit and member of the Tulalip Tribes with a BFA from the University of Washington (2000). Together with his uncle, he is known for the massive sculpture fronting the Tulalip Casino. His steel sculpture in the garden, "Ancestors Moving On," assembled from 15 joined, carved steel circles, swings in the wind, with his aspiration "to teach others of the lands that [his] own people used to walk."
There are other notable sculptors: Ethan Stern brings blown and wheel-cut glass pieces that exhibit subtle marks of carving; their transparent surfaces reveal complex opaque swirls within. The Iraq-born Sabah Al-Dhaher has carved human forms in stone: his "Winged Goddess" lacks wings and is ominously hollow, indicating the disturbing condition of mankind.
Also commenting on the human condition are Phillip Levine's six sculptures in bronze, steel, walnut and sand. "Far Enough" channelsGiocometti. In "Fear and Habit," eight homunculi posture precariously on platforms surrounding a cage.
If this take on mankind appears too grim, glance at Sue Roberts' cartoonish assembly of "The Gunn Family." Oh, no—Dad, Mom and kids are all packing heat!
You could relax among the subtle shading of Richard Nash's classic abstracts: acrylics, looking like charcoal drawing. They would grace any living room.
And what's this? Karla Matzke (yes, her again) has created three astonishingly sensual nudes in pastel: very realistic without appearing coldly photographic. In "Spooning" it feels like something very private is about to happen. How does she get her models to hold still so long when the moment captured is so thrilling?
More Visual...
A magical resonation
In "Evidence," the current show at Smith & Vallee Gallery, two exceptional artists give us their views—David Blakesley, of a world that might have been or what it might become, and Kathleen Faulkner, images of the forest, examined close-up and transformed by the creator's eye.…
Native Son
The life and art of James T. Pickett
Pencils and paper were scarce commodities on the remote Mason County homestead where James Tilton Pickett grew up, but that didn't stop him from drawing.
Instead of filling sketchbooks and stretched canvasses, he committed his lines to a variety of repurposed barnyard materials. Charcoal…
Creative Control
Logan McQuaig goes solo
Among the eye-catching paintings Logan McQuaig will be debuting at his upcoming "Creative Control" exhibit are a stern buffalo with a bird on its shoulder and a gun held between its hooves ("Stand Your Ground"); a hapless guy munching on a McDonald's Big Mac while a nuclear bomb lights up…


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Judge to rule on forcing alleged dog hoarder to relinquish animals
March 31, 2014
A Nevada County Superior Court judge has been asked to rule on an unusual motion that would force an alleged dog hoarder to relinquish six animals — even though his case is currently wending its way through the court system.
Gary Dean Perkins, 53, had been arrested in November on felony animal cruelty charges after Nevada County Sheriff's Animal Control officers seized 31 dogs from a parcel in the 16000 block of Bear Trap Springs Road, where Perkins reportedly was squatting.
The dogs ranged in age from 6 weeks old to adult and reportedly were emaciated, with no food or water found. Deputies reportedly also found five dead dogs on the parcel, and necropsies revealed death by starvation. One dog was pregnant, driving the total number of live dogs involved in the case to 41.
All but six of the dogs seized have been adopted, according to Cheryl Wicks at Sammie's Friends animal shelter.
The remaining animals cannot be rehomed until the court sorts out the case against Perkins, who reportedly has asked to get those dogs back.
Sammie's Friends has been caring for those six dogs since Nov. 23, at a cost of $12 a day each.
Recommended Stories For You
And that's not counting the cost of food as well as the medical care they needed, Wicks said.
Of the 17 adult dogs seized, 10 had heartworm — and all the dogs needed vaccines, spay and neuter services, and other medications.
Wicks estimated the total cost of treatment to be in excess of $8,000.
In court March 13, Judge Candace Heidelberger addressed the motion filed by Deputy District Attorney Jim Phillips to confirm the allegation charging animal cruelty and to request an order relinquishing ownership of the dogs, and impose the costs of care.
Phillips said a fairly new section of the California penal code affords a mechanism by which the government can seek seizure and forfeiture of dogs and cats under some circumstances, based on an unacceptable level of care.
Deputy Public Defender Micah Pierce argued that his client should be allowed to retain ownership of the dogs, pending any legal order — which he said could not exist without a conviction.
Heidelberger, however, said she was unsure if there had to be a prior conviction or whether there could be another basis for forfeiture.
"Sammie's Friends has a lien on animals that exists even if he is acquitted, in excess of $8,000," Phillips said. "That must be paid before the animals can be turned over to the defendant. If he is indigent, how on earth is he ever going to be able to pay for these animals, much less care for them?"
In the end, Heidelberger said there was a right by the court to order the forfeiture without a criminal conviction.
She set a new hearing date for that motion, as well as a preliminary hearing into the evidence, for April 10.
Pierce advised the court he would be filing a motion to dismiss the charges as overbroad, saying, "There was no intentional abuse. He ran out of money to feed them."
That motion was set to be heard April 3.
To contact City Editor Liz Kellar, email firstname.lastname@example.com or call 530-477-4229.
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Trending In: Local News
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- Never again: Nevada City filmmaker produces series on incarceration of Japanese during WWII
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- 14,000 pounds of trash pulled from Nevada County waterways by nearly 900 SYRCL volunteers


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Jr Doctors strike spreads from Telangana to AP - Tv5
Sat, Nov 22, 2014, 07:19 AM
Jr doctors in Telangana are striking since many days and the High Court also instructed them to call off the strike. Now their AP counterparts are taking the path of Telangana Jr doctors as their talks with the Government have failed. The AP Jr doctors announced that they will sit on strike and all the services will be called off except emergency services. Watch the video for more details of this news .. . . . . .


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Tweets from @TBHomeTeam/writers
MORE from our HomeTeam writers.
Wed. May 22, 2013 | Joel Anderson
FORT MYERS — The breakthrough seemed only a run away, especially after Daniel Portales delivered the tying single in the bottom of the fourth.
Then Jesuit failed to score with the bases loaded. It happened again with runners at first and third base in the bottom of the fifth. In the next inning, the Tigers stranded a runner on second base.
They were running out of chances, and nearly everyone in their dugout and JetBlue Park knew it.
"I kept saying, 'This is our inning. This is our inning,' " Portales said. "But it didn't happen."
Ponte Vedra eventually took advantage of the opportunities that Jesuit wasted, scoring on Chris Levitt's RBI single in the eighth inning for a 2-1 Class 5A semifinal win Wednesday night.
It was a shocking finish for the Tigers (27-3), which came into the game as MaxPrep.com's fifth ranked team in the country and a prohibitive favorite to return to the championship game.
Instead, Jesuit was again denied a state title in its fourth consecutive trip to the final four. The loss was particularly ironic since it came against a team the Tigers defeated 19-0 in their 2010 semifinal match — the first of their recent state tournament teams.
"It's been four years now and we just can't come through," said Portales, a senior and USF signee. "But one year we're going to get it."
Because, of course, there's always next year to look forward to on North Himes Ave.
But this team was supposed to be different. The Tigers had seven college signees on their roster, they returned a large senior class with plenty of big-game experience, and they boasted so much talent they were the nation's consensus No. 1 team much of the season.
In retrospect, there should have been some signs Jesuit wasn't quite as unbeatable as it seemed.
The Tigers averaged fewer than four runs in their final eight games, a huge dropoff from their 12 runs-per-game average in their first 22 games — most of them lopsided wins.
Their offensive struggles showed up again against Ponte Vedra (20-11), which went with 6-foot-3 senior right-hander Matt Deegan to start the game.
Jesuit could only muster one hit against Deegan in the first three innings, while Portales also managed to keep the Sharks — who entered with their top hitter averaging .275 — off the board.
In the fourth, Ponte Vedra scored its first run on a throwing error to first. Jesuit sophomore Ronnie Ramirez came in for Portales after that and got the final out of the inning.
The Tigers tied the score at 1 on Portales' RBI single, which brought Leland Saile home, in the bottom half of the inning but failed to score again with the bases loaded. It would become an ominous trend.
"We got close," Jesuit coach Richie Warren said. "We had a lot of opportunities tonight."
The Tigers never got a runner past second base after the sixth inning, and Ponte Vedra finally made good on one of its scoring opportunities in the top of the eighth against Jesuit ace Spencer Trayner.
Trayner, who came in for Ramirez after Jordan McKenna's leadoff single for the Sharks, got off to a rough start: his balk moved a runner over to second; a wild pitch allowed the runner to get to third; then he surrendered the go-ahead RBI single to Chris Levitt.
Trayner quickly got the next three outs but Jesuit, once again, couldn't generate any offense in the bottom half of the inning and went down 1-2-3 to bring an ignominious close to the season.
The Sharks, now in their third final-four appearance in only five years, took particular pleasure in delivering a knockout blow to the team that had embarrassed them only four years earlier.
"That was the best loss of our program," Ponte Vedra coach Tom Stanton said. "It was an eye-opening experience. We learned from that, and we weren't scared to see them."
Joel Anderson can be reached at firstname.lastname@example.com or on Twitter @jdhometeam.
Photo: Daniel Portales reacts after Jesuit's 2-1 Class 5A semifinal loss to Ponte Vedra. MARC BEAUDIN | Special to Times


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By Varietal
A bison burger topped with grilled wild mushrooms, wild blackberry marmalade, and edible flowers served on a buttermilk biscuit.
Fast and easy to prepare, this combination of flavors will leave your mouth wanting a second burger!
Greetings! This is a special recipe that I have made for my family and friends. Believe me, they are picky eaters and I think you'll find that this is a delicious recipe.
This burger is made from an international blend of spices and ingredients (Italian, Indian, and Mexican) that is sure to make your mouth water!
Introduction: Sauerkraut was one of my favorite foods when I was young and each summer I would usually make about 15 gallons which lasted most of the year.
My Mom has been dating an Australian man for the past 13 years and every time they come to visit in the US (his job has them living in Sweden for years at a time), they both crave a good old fashio
This recipe celebrates the richness of the Pacific Northwest, from our thriving coffee culture to our commitment to great tasting, locally grown natural produce and meats.
In Dallas, there are many versions of a "Cowboy Burger".
This burger is a bit country. We live on a ranch where we raise beef cattle and Boer Goats(chevon). I thought it was best to share with you my idea of the best of both worlds.
Butter Burgers are a Wisconsin classic, where a burger is topped with big jolt of pure butter. I wanted to update the concept a little and make it more contemporary.
This burger takes the flavors of the traditional American Thanksgiving and puts it in a burger to be enjoyed all year long!
The ingredients used in this recipe are grown on the Prairie in North Dakota, near my home in Minnesota.
Seattleites are proud to claim Jimi Hendrix as one of their finest exports, long before a string of latte stands put us on the map.
I have been working on a burger that is so wonderful! that, I've got share it with you. It is full of a passion, that transcends the shores into the ocean and back to your palate.
I have been making the BBQ sauce for 3 yrs now and everyone loves it, so I decided to incorporate my sauce in a burger. So far, it has been a great success.
This is a great local American burger because throughout Wisconsin, burgers and brats sizzle side-by-side on the grill at tailgate parties and picnics and I have also known them to sizzle during a
Colorado is a state full of breathtaking natural beauty that encourages a healthy and active lifestyle. This burger uses grass-fed beef raised right here on the plains of Colorado.


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PRZOOM - /newswire/ -
Windsor, VT, United States, 2012/02/01 - Pittsburgh web design company ProFromGo announces the launch of its long-awaited content development services - ProFromGo.com.
Pittsburgh web design firm ProFromGo announced the official launch of its content development services division this week. The announcement comes after nearly 6 months spent creating a Content Strategies and Solutions team to round out the design firm's stable of internet professionals.
RJ Hudson, chief writer for the firm, stated that the culmination of this project was a natural integration of web design and content development:
"You cant have one without the other a beautiful website that provides no value in the form of content is doomed to rapid failure. People can't just see your website. They have to be compelled in some way, and this is accomplished through high-value content."
Content on the web consists of web pages, articles, songs, files, movies, videos, and more. Consumer demand for content grows exponentially each year - in fact, according to the Pew Internet and American Life Project, 78% of American adults used the internet in 2011, and nearly all of them were in search of some type of content.
Search industry experts have long touted the critical need to integrate web design with content development, but few companies have been able to successfully integrate these services because they are so vastly different. But Chris Vendilli, Founder and Chief of Operations for ProFromGo, says that these differences can be successfully integrated:
"That's like saying an engineer and a salesman are so different that one single company can't both make and sell cars. That's just not reality. We've developed our content division because we have an intrinsic understanding that a website must have two elements in order to be successful: powerful design and engaging content. They're inseparable, and we've now brought all of this in-house."
The Pittsburgh firm has been recognized for its award-winning website designs and reports that its content development division which produces everything from articles to sales pages consists of master writers and a multi-level editing system.
However, some in the industry have complained that search engines are unfairly forcing businesses to develop content.
"That's another SEO myth." Says Vendilli. "Businesses don't have to become content farms, but they do have to offer content that is relevant to their products or services and that content needs to be of the highest quality. After all, how else are you going to sell yourself? Proper web design appeals to a consumer visually, mentally and subconsciously, and compelling content appeals to a consumer emotionally. When you put all of this together, you can create a website that rockets its way to success."
ProFromGo (profromgo.com) is a local Pittsburgh web design and content development firm with a powerful international reach.
Agency / Source: Content for Conversions
Availability: All Regions (Including Int'l)
Traffic Booster: [/] Quick PRZOOM - Press & Newswire Visibility Checker
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my boobs are very far apart and have a huge gap between them. When ever my pectoral muscle tenses they deform. There is skin laxity and they just look terrible. I have tried to take some pics myself. The quality of the pics is not great and my mirror is filthy I am sorry. My current implants are round smooth mentor under the muscle saline. They are a little over 10 years old.
I had an initial consultation a few months ago and am scheduled to meet the doctor on Monday to talk about my options. I'm hoping to maybe go over the muscle this time. They want me to tell them how many cc's I want but I have no idea. I don't want clown boobs, but I want them to fill up the loose skin.


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Services on Demand
Related links
Gaceta Sanitaria
Print version ISSN 0213-9111
COLOMER-REVUELTA, Concha et al. Child health. Gac Sanit [online]. 2004, vol.18, suppl.1, pp.39-46. ISSN 0213-9111. http://dx.doi.org/10.1590/S0213-91112004000700009.
Children in Spain are considered as being a population group with few health problems and needs therefore making it almost invisible in research and services' planning. Generally, it is not taken into account that this is a development period with very high vulnerability to physical and psychosocial risks whereas there is a positive response to protective factors. This article covers some data and thoughts on their health problems that are improving (mortality, cancer), those that persist or worsen (environmental quality, abuse, mental health, obesity, disabilities and lifestyles) and new challenges relating to quality of life and gender and social class equity. Responses provided by the health services are reviewed, as are environmental policies and health promotion and specific care is proposed for boys' and girls' health from a children's human rights-focused perspective.
Keywords : Childhood; Boy; Girl; Gender; Inequalities; Health determinants; Quality of life; Human rights; Environment.


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Afro-American Studies Program and the Center for the Study of Black
Literature and Culture have merged into the newly created Center
for Africana Studies.
Center for Africana Studies is a space for "the critical examination
of not only the cultural, social, political, economic and historical
experiences of African-Americans, but of the African Diaspora,"
said Dr. Tukufu Zuberi, who has headed the Afro-American Studies
Program and is now the director of the new center.
Tuesday, President Judith Rodin, Dr. Zuberi, SAS Dean Samuel Preston
and others welcomed four new faculty members (Michael Eric Dyson,
Leslie Callahan, Timothy Rommen and Cheikh Babou) and invited guests
to the official opening of the Center located in Suite 331A, 3401
Walnut St.
Center is a mecca for research projects, fellowships for emerging
and established scholars, publications, conferences and working
groups. The Center continues to promulgate the mission of the Afro-American
Studies Program by offering a major and minor in African-American
Studies, and producing cutting-edge programming that includes public
conferences, lectures, book talks, and forums," said Dr. Zuberi.
Program, one of the nation's oldest research programs dedicated
to the study of Africa and the African Diaspora, was founded in
1972. It was established in response to a national movement by college
students across the U.S. to add courses exploring black history,
literature and culture to their school's curricula.
Preston noted that "Penn's legacy in Afro-American studies
reaches back much further than 30 years, to the late 19th century
when W.E.B. Du Bois wrote The Philadelphia Negro while affiliated
with Penn. In 1898 he wrote that "It is to the credit of the
University of Pennsylvania that she has been the first to recognize
her duty and in so far as restricted means and opportunities allowed,
has attempted to study the Negro problem in a single definite locality."
Dr. Preston went on to note that Du Bois was in fact denied
formal faculty status and that it would be 70 years before the first
Afro-American studies courses were taught at Penn. He concluded
his remarks by suggesting that "we have acted upon Du Bois'
notion of a "single definite locality" for Afro-American
and Africana studies at Penn. With the talented faculty brought
together under the aegis of the center, the sky is the limit."
Program has experienced major expansion in the past few years with
Penn's renewed commitment to the recruitment of African-American
Studies faculty. Dr. Rodin noted that the recent faculty hires "strengthen
an already formidable lineup of scholars from a wide range of disciplines."
She also added that "we need just the kind of path-breaking,
interdisciplinary scholarship in Afro-American studies that is taking
place at Penn now." The establishment of the Center will "capture
these synergies to enrich our understanding of all the dimensions
of the African American experience. I wish to stress my belief that
all literary genres, all artifacts, and all
forms of cultural expression are fertile ground for "serious
Center will mark the 30th anniversary of African-American studies
with Back to the Future of Civilization, a year-long series
of events, including moderated panel discussions on literature,
society, arts and culture, critical theory and history in the African-American
and African Diasporic experiences. Issues in Black Independent
Cinema: The Documentary begins at 7 p.m. tonight at DRL. Reservations:
call (215) 735-3785.
Almanac, Vol. 49, No. 6, October 1, 2002


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The Presidential campaign of 1828 was brutal. Andrew Jackson felt that the election of 1824 had been stolen from him (he won the popular vote) and John Quincy Adams was defending his position as incumbent. Although it was considered undignified for the candidate to campaign, their supporters pulled out all the stops even attacking the wives. Louisa Catherine Adams was considered by many a foreigner who brought unwelcome influence to Washington, (she was born and raised in England, although her parents were Americans), but the worst criticism was reserved for Rachel Jackson. Called a whore, a bigamist, and an unsophisticated westerner (my goodness she smoked a pipe!), many considered her unqualified to inhabit the White House.
When Rachel Donelson was 12 years old, her family left Virginia to travel to the frontier of Tennessee. The trip was arduous and the situation with the Indians unsettled, but eventually they established themselves as one of the most prominent families in the area. They built a palisade which surrounded the family home and several cabins where boarders lived. This is where Rachel and Andrew met and fell in love. Rachel was a beautiful young woman and Andrew was tall and handsome with an instinct to protect women. Rachel may have seemed like she needed protecting – from her husband, her first husband.
A few years earlier, when the situation with the Indians was very dangerous, the family briefly moved to Kentucky. There Rachel met and married Lewis Robards. The Donelsons and Robards were of similar social standing and circumstances. Lewis was from a large family who welcomed and liked Rachel. It seemed like a good match. And it was for a while, but the vivacious and friendly nature of Rachel that attracted Lewis became a problem. She was raised with seven brothers and quite innocently enjoyed the company of men, but Lewis was of a jealous nature. By 1788, the situation had become unbearable and Rachel's brother came to get her, bringing her home to her family in Tennessee. It is unsure whether Rachel intended the move to be permanent or not, but later in the year Lewis followed her to Nashville and moved in with her and her family.
It was during this time that Andrew Jackson moved to Nashville. Rachel and Andrew were attracted to each other, but there is no evidence that their relationship was inappropriate. However, Lewis couldn't accept this; he forbid her to talk to Jackson and interrogated her about the few interactions they had. Finally, during the summer of 1789, Lewis made the mistake of telling other men that Jackson was too intimate with his wife. Lewis was not well-liked, but Jackson and the Donelson family were. Word got back to Jackson and he confronted Lewis and threatened to cut off his ears.
Rather than challenge Jackson to a duel, which he undoubtedly would have accepted, Lewis had a warrant issued against him. While traveling to see the magistrate, under armed guard, the looks that Jackson gave Lewis terrified him so much that he ran off. Of course the fact that the guards allowed Jackson to carry a knife and to chase after Lewis, may have had something to do with the fact that Lewis didn't show up at the magistrate's office. Without a complainant, the charges were dismissed. This was the last straw for Lewis, he left Tennessee and returned to Kentucky swearing to have nothing more to do with Rachel and her family.
By this time, Rachel and Andrew knew that they were in love and wanted to marry. While attitudes toward marriage and divorce were more relaxed in the west, including self-marriage and self-divorce determined by the families rather than a bureaucrat from across the mountains, the Donelsons liked to abide by the legal niceties when possible. This was particularly important with regard to property rights. Rachel's father had died in 1885 and although his will had yet to be settled, anything that came to Rachel would legally belong to Lewis Robards.
Women had no legal right to divorce (at least none that would be acceptable to a southern judge), so the hope was that Lewis meant what he said and would file. In the meantime, the only option Rachel and Andrew had to live as husband and wife was to elope to an area beyond American Law. The closest place whether Andrew could hope to earn a living was Natchez (now in Mississippi) which was then still Spanish territory. In the summer of 1789, Andrew began to establish connections there.
That fall, they heard rumors that Lewis was angry and preparing to come get Rachel and force her to return to Kentucky with him. He was within his legal rights to do so and Rachel and Andrew made a decision. In December of 1789, they boarded a flat boat and headed down the river. There is no documentary evidence that they married in Natchez, but there is evidence that they presented themselves as Mr. and Mrs. Jackson. They made connections with family friends and entertained, making no effort to hide who they were. Neither purchased land; presumably they rented with the intent of returning to Nashville when they could.
By July 1790, Lewis had not returned to Nashville and Rachel and Andrew decided to go home. The journey up Natchez Trace with an armed party was uneventful with one exception. One of their traveling companions knew Lewis Robards and later reported to him that Rachel and Andrew were living as husband and wife. This testimony was the tangible evidence he had been waiting for and he made a petition to the Virginia legislature for an action of divorce. This gave Lewis permission to sue Rachel for divorce in Kentucky supreme court. (Kentucky was still part of Virginia at the time.)
There were other requirements that had to be met: notice in the Kentucky Gazette, filing a writ in Kentucky, and taking depositions from witnesses. But, Lewis put off doing these things. Maybe because of anger toward Rachel or because of lingering hopes of receiving her inheritance, nevertheless, the divorce wasn't finalized until three years later in September of 1793. The charge was adultery, but neither Rachel nor Andrew appeared in court. After all, they weren't going to fight something they had been hoping for all along.
Rachel and Andrew had been living in Nashville as husband and wife since 1790, so they quietly married (remarried?) on January 17, 1794. They enjoyed their lives together and the events of the previous years receded into memory until the election of 1828.
In spite of the ugliness of the campaign, Jackson won the election by a landslide. Briefly, Rachel considered not going to Washington with Andrew, but they decided that that would be admitting they were wrong, which they didn't believe. So Rachel prepared to become First Lady of the land. Sadly, it wasn't to be. On December 22, 1828, Rachel Donelson Jackson died, leaving Andrew a deeply sad and embittered man. He would always blame those who attacked Rachel for her death. Two days later, Rachel was buried in the white satin gown she had planned to wear to the inaugural ball.
A Being So Gentle: The Frontier Love Story of Rachel and Andrew Jackson by Patricia Brady
C-SPAN: First Ladies Influence and Image – Rachel Jackson


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Buy a Print
'New morality; - or - the promis'd installment of the high-priest of the Theophilanthropes, with the homage of Leviathan and his suite' (includes James Maitland, 8th Earl of Lauderdale; Sir Francis Burdett, 5th Bt; Michael Angelo Taylor; Charles Howa...)
by James Gillray, published by John Wright
etching, published 1 August 1798
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Closing Bell For The Lincoln Town Car
by Gunnar Heinrich ::: img Ford Motor Company ::: 2011 Lincoln Town Car
LINCOLN'S Town Car is based on a platform so old, it has several mentions in the Bible spans three decades, six US presidencies, and plenty of upticks and downturns in the economy.
Of course we're talking about the perennial Panther platform; that 70's design that's been FoMoCo's staple workhorse for full size rear wheel drive sedans since Chaka Khan first sang I'm Every Woman.
The platform for every Crown Victoria, Grand Marquis, and, yes, the Town Car is due to sunset next year and with it the Town Car's Canadian assembly plant at St. Thomas, ON.
The closing marks closure to an era for the cop cars, taxis, and limos that dominate US city streets – New York and Washington, especially. Bankers and politicos and their respective drivers will need to adjust.
Change was long overdue.
Lincoln's Town Car hasn't seen a real update to its current – admittedly handsome form – since 2003. The 2011 model entices the would-be buyer with scant few options – which includes basic features like HID headlamps that other rivals now sell as standard equipment at the $45K price point. Depreciation, too, is steep.
But for every ending there ought to be new beginnings and yet it's unclear whether Lincoln has a suitably large replacement for the Town Car in the pipeline and, no, the MKS doesn't count.
Despite popular belief, we still like'em big, plush, and roomy here in the land of the free-to-roam. Why else would more American fleets use Town Cars than any other car? According to the Times, three quarters of all New York livery services use Lincoln's capacious flagship.
Panther flavored food for thought.


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June has arrived and with it a multitude of festivals. If you can't make to one, then you're just not trying hard enough. Here are options for your next four weekends. Click on any of the festivals to see full lineups, more ticket options, and info on camping.
Weekend 1: June 5-7
Your first choice is between jam banding out in the Ozarks of Arkansas or head banging with Kid Rock down on the infamous Redneck Riveria.
Wakarusa Camping and Music Festival
June 4-7, Mulberry Mountain, AK
$159 for a full four day event pass (doesn't include camping)
Headliners: Black Crowes, Gov't Mule, Galactic, Yonder Mountain String Band, Les Claypool, G. Love
June 4-6, Enterprise, AL
$150 for three day pass
Headliners: Kid Rock, Brooks & Dunn, Alan Jackson, Taylor Swift, Randy Owen, Charlie Daniels Band, Drive By Truckers
Weekend 2: June 12-14
Of course Bonnaroo is the music festival to end all music festivals, but if you don't have the coin to get there, consider checking out the smaller Harmony Festival in Santa Rosa. Or if full on country is your thing, swing through downtown Nashville.
June 11-14, Manchester, TN
$249.50 for a four day pass includes camping and parking
Headliners: Bruce Springsteen and the E Street Band, Phish (2 Shows), Beastie Boys, Nine Inch Nails, David Byrne, Wilco, Al Green, Snoop Dogg, Elvis Costello, Erykah Badu
Harmony Festival
June 12-14, Santa Rosa, CA
$30 advance single day / $40 at the door
Headliners: Michael Franti & Spearhead, India.Arie, Cake, Dead Kennedys, Matisyahu, The Orb
Country Music Festival
June 11-14, Nashville, TN
$175 for a four day pass
Headliners: Trace Adkins, Brooks & Dunn, Little Big Town, Josh Turner, John Rich, Lee Ann Womack, Darius Rucker
Weekend 3: June 19-21
It's all Canada all weekend long. Alternative bands take over at Toronto's version of SXSW while Virgin kicks off their first of five Canadian festivals.
North by Northeast
June 17-21, Toronto, ON
$50 for a five day wristband
Headliners: Black Lips, Burning Brides, Wintersleep, The Sonics, Youth Brigade, The Cool Kids, GZA
Virgin Festival: Montreal
June 19-20, Montreal, QB
$75 for a single day ticket
Headliners:  Black Eyes Peas, Simple Plan, Akon, The New Cities, Hedley, New Kids On The Block, Lights, Live
Weekend 4: June 26-28
10 days of music. Over a million people. The biggest music festival in the midwest that you may not know about kicks off  the last weekend of June.
June 25-July 5, Milwaukee, WI
$15 single day ticket (additional cost to see headliners)
Headliners: Bon Jovi, Keith Urban, Bob Dylan, No Doubt, Kid Rock, Lynyrd Skynyrd, Puddle of Mudd, Willie Nelson


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Ramp ceremony a final salute for fallen
Focus on returning comrades' remains home as soon as possible
By KEVIN DOUGHERTY | STARS AND STRIPES Published: May 30, 2005
KIRKUK, Iraq — A soldier died the other day in Iraq. He was in his early 30s, from Tennessee. The death came in a vehicle accident. His identity is not the point of this article.
More than a dozen soldiers perished last week, and over 1,600 others have preceded them since the beginning of the Iraq war in March 2003. Every time a servicemember dies in Iraq, no matter the circumstances, the Army and Air Force conduct a "ramp ceremony" to repatriate the remains.
What follows is an account of one such ceremony:
Soldiers learn to wait. They really have little or no choice. In Iraq, that's not the case when troops die. The aim, according to one Army officer, is to have the servicemember's remains on a plane and heading home within 24 hours.
Usually, the military makes it happen in about half the time.
The soldier being sent home today awoke on the day of his death in time to see the morning sun. Just before sundown the same day, several hundred troops gathered in his honor to pay their respects and offer a farewell salute.
They came to Kirkuk airfield wearing helmets and battle dress uniforms and soon fell into formation. Few, if any of them, knew him personally. He served in a different brigade, but they came. The mood is somber. There is no pomp, no band, only a unit flag and Old Glory.
After the C-130 lands, it taxis toward the soldiers, and the formations tighten. Except for the aircraft's engines, there is silence. The distance between the plane and the troops is about 100 yards.
Two forklifts pull up to the rear of the aircraft, which now faces the ensemble. The belly of the plane surrenders cargo pallets and 12 troops, who head for the passenger terminal. The engines stop.
From the nose of the aircraft, the sun is slightly off to the right. Pollution from burning oil wells gives the sky a hazy, yellowish-orange hue, and the sun slips ever closer to the horizon.
An airman grabs a broom from inside the fuselage and sweeps the back of the ramp. Another airman exits the plane, bows forward and splashes water onto his head. For U.S. forces, Iraq has been a baptism of faith — faith in themselves, faith in their fellow soldiers, faith in their country and its ideals.
On command, the assembled soldiers begin to march toward the rear of the plane, some formations four abreast, others more. They create a corridor and wait.
The refrain "present, arms" echoes among the ranks, and all salute. An Army ambulance pulls up to the far end of the corridor. The salutes end, but only for a brief moment as the casket is removed from the back of the wagon.
A slight breeze kicks up. The sun progresses ever so close to settling down for the night as the escort nears the back of the aircraft.
Chaplain (Lt. Col.) John Worster leads the procession. Eight soldiers carry the casket, with a ninth and tenth positioned at the head and feet. Capt. James "Cole" Packwood and 1st Sgt. George Laubhan, both of the Headquarters, Headquarters Company, 116th Brigade Combat Team, follow them.
It's their first time serving as escorts.
"This is probably one of the proudest things I've ever done," Packwood says later. "Even though it wasn't one of our soldiers, it felt like it."
After a soft prayer and hymn, the casket is slowly and solemnly carried onto the plane. The escorts re-emerge and depart the scene.
"All secure, loadmaster," Packwood says, before he and Laubhan do an about-face and leave the rest to the Air Force.
Commands are given, the formations file back toward a hanger, soldiers disperse. The scene slips away in an instant, much like the setting sun now hugging the horizon.


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Chocolate Covered Cherries are one of those treats that you can never go wrong with. It's got the perfect combination of sweet and savory. There's something about the mixture of chocolate, cherries and a sweet milky center that go perfectly together. The past few years I have started to test my candy making skills and in the process whipped up this Chocolate Covered Cherries Recipe. These are so easy to make and in my opinion taste so much better than the store bought kind. I hope you enjoy.
Chocolate Covered Cherries Recipe
What you need:
- 1 Jar of Maraschino Cherries with Stems
- Cherry Juice from the Jar or Cherries
- 1 Package of Chocolate Chips (Milk, Dark or Semi-sweet)
- 3 Cups Powdered Sugar
- 1/4 Cup Melted Butter
- 4 tsp of Vanilla
- Dash of Salt
- 1/4 Cup of Sweetened Condensed Milk
You will also need mini baking liners. These are the ones that I used and they work great.
Make Chocolate Covered Cherries Homemade!
Before you make this recipe you want to make sure that the cherries are dry. So the night before drain the cherries and pat them dry with a paper towel. Place in the fridge and they will be ready to go by morning. Be sure to reserve the jar of cherry juice to use with the chocolate covering.
Mix the butter, vanilla, salt and sweetened condensed milk together in a small bowl Pour this mixture into the powered sugar and mix well. Place this fondant mixture in the fridge for an hour to chill and harden.
OR You can SKIP This step and purchase already made Vanilla Fondant which tastes great!
At the same time place your dried cherries into the freezer for an hour to harden up. This makes them easy to wrap the fondant around.
After about an hour take your fondant and cherries out of the fridge/freezer. Take about 1 teaspoon of the fondant and wrap around the cherry tightly. To smooth it out you can roll it in your hand, just make sure that the stem doesn't fall off.
Place all of the fondant wrapped cherries into the freezer for about an hour.
After an hour melt your chocolate chips in the microwave, adding the reserved cherry juice as needed to make it thin enough to dip the cherries.
When the chocolate is done, take your frozen fondant cherries out of the freezer. Dip them one at at time into the chocolate mixture making sure you cover them entirely.
Once they are covered raise them over the bowl and let the excess chocolate drip off. Place them on wax paper.
After your all done place the cherries into the fridge and let harden. You can eat these cherries right away but the longer you let them sit the creamier they will be in the center. Place in an airtight container in the fridge for up to 3 weeks.
Making this chocolate Covered cherries recipe is super easy and you will be so glad you did. They taste great and make a wonderful addition to any occasion. If you put these in a nice little gift box they would make the perfect gift for Christmas, Valentine's Day, a birthday or just because.
If you need more ideas on Valentine's Day Gifts Here are some of my favorites.


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This step is elective but will greatly multiply your chances of getting interest from other members on the website. Write a little something about yourself, your interests and attach any nice photographs of yourself.
Utilise our website's advanced search to encounter members who fulfill your requirements. Or just leaf through listings and encounter members that you find attractive.
Discovered someone who appeals to you? Drop them a line or just a wink and find out if they're interested in you. With a little luck you'll be on your way to a lifetime of happiness.
You won't regret it


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Michelle Obama shops at Target, becomes Deal Diva in Chief
Let's all have a moment of appreciation. First lady Michelle Obama secretly shops at Target. If she's anything like all other Target shoppers, she probably ran in for a tube of Crest and left with 18 wall sconces and three pair of shoes. Good sleuthing, Associated Press!
Photo: First lady Michelle Obama, wearing a hat and sunglasses, leaves a Target store in Alexandria, Va., Thursday after doing some shopping. AP.


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Honda Insight Hatchback 2012 for sale
345 Views
Black 5 Door N
Translate this page :
- Vehicle
- 2012 Honda Insight
- Body
- Black 5 Door Hatchback
- Engine
- 1300cc Automatic Hybrid Engine
- Odometer
- 55,000 kms
- Safety Rating
ANCAP Safety
Based on rating for 10+ models with frontal+side+head airbags
- Fuel Economy
- Yearly Cost: $ 1290 (source: Fuelsaver)
Seller Details
- Seller:
- Private Seller
- Location
The Honda Insight is a hybrid electric vehicle, saves a lot of fuel and great pick up makes it a unique car. The Insight is the 1st production vehicle to feature Honda's Integrated Motor Assist system. The Insight has an official CO2 emissions figure of 80 g/km which is still the lowest in the market with 4.4Litre/ 100 km of fuel efficiency. It has great factory fitted speakers, GPS inbuilt, bluetooth, radio not in Japanese, AirCondition and manual heater. The best option are Cruise control which gives a better fuel efficiency if opearted systematically. Stops Idle on red light if stationery for more than 3 seconds. In addition, ECO ASSIST which includes a dedicated ECON button that enables the driver to initiate a range of functions that increase the fuel economy of the IMA system via a single button press. In ECON mode, the driver trades off a measure of performance for enhanced fuel economy but gains some advantages like
1. Optimizing throttle angle input and CVT operation.
2. Limits power and torque by approximately 4 percent (full responsiveness is provided at wide-open-throttle)
3. Increases the potential for engaging the Idle Stop feature sooner.
Had a service done recently and has factory fitted tyres too.


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A TEENAGE driver arrested for allegedly doing burnouts and trying to outrun police in the early hours of Sunday has joined friends on Facebook to mock police.
Koen Attard, 19, was photographed by the passenger who was with him about 2am on Sunday, at Direk, in Adelaide's north, when it is alleged he was doing burnouts in his silver Ford sedan, before fleeing from police and crashing into a gutter.
The picture, showing Attard handcuffed and in the back of a police car, has been turned into memes mocking the police and posted on Attard's Facebook page and other car club member sites.
Attard had commented on the posts, saying he told the arresting police officer he had tried to outrun them because he thought he would win, but the comments were removed soon after The Advertiser raised the memes with him.
Attard, who has been charged with driving in a dangerous manner and failing to stop, told The Advertiser he had nothing to do with producing the memes and denied he was mocking police over the serious incident.
"There's nothing to mock the police about ... they are doing their job," he said.
"I didn't want anyone really knowing about (the incident) ... but I know some photographs have been posted."
Attard said his arrest came after up to 80 cars had gathered in the area as part of a car cruise. "Everyone got out of there pretty quick when the cops came," he said.
Police will allege that about 2am on Sunday, they were called to Edinburgh Road after reports of several cars doing burnouts.
Police briefly pursued a silver Ford sedan, before it collided with the gutter and came to a stop.
Attard, from Plympton, was bailed to appear in the Elizabeth Magistrates Court at a later date. His car was impounded for 28 days.
"With the Clipsal 500 starting this week, this is a timely reminder to all road users to drive responsibly," a police spokesman said.
Attard said he would not take part in any hoon driving in the future. "I'll go to the track if I want to do anything like that," he said.
Originally published as Teen hoons mock cops on Facebook


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Anxious but Excited - Houston, TX
Going in tomorrow afternoon so anxious!! A little scared also.. Having procedure done in Houston.. I'm 43 years old had previous breast Aug.. Decided just last month I wanted the BBL.. I'm only 130lbs so hoping enough fat can be harvested from the areas to get a good result. I have everything packed and ready I live about hour half away from Houston. A little nervous about not being able to... READ MORE
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Thank you Destiny81, my jeans don't fit can't button most of them, but I do have a couple pair that were fitting big that fit perfect now!! They are Miss Me Jeans and fit well.. It's getting Beyter and better!! READ COMMENT


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Knife Event, Grape Festival, First ThursdayPosted by admin |
All events listed in 8 Days a Week are open to the public. To submit a listing, send to Fayetteville Free Weekly, P.O. Box 1607, Fayetteville 72702, or e-mail email@example.com at least two weeks prior to the event. Submissions are printed as space allows. Photographs and art are also considered.


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She refuses to go back to her doctor or do anything about this. She has not lost any weight, and says" think how big i would be if I didn't have it". I am very worried about her, I know that vomiting that much every day for five years cannot be good for her. She will not listen to my pleads for her to have it looked at. Any suggestions on what could be wrong with her lapband? and any scare tactics I could use to get her to go to the doctor.
My Sister Had Lapband Surgery About 5 Years Ago and Has Pretty Much Been Vomiting at Almost Every Meal She Eats Since Then?
Doctor Answers 1
Vomiting after Lap Band
I appreciate your question and your concern for your sister. Let me assure you that vomiting after a gastric band or for that matter, after any weight loss surgery is not normal. She may have a slipped band, malpostioned band, overfilled band or other problems with the band. You should not use any scare tactics, but rather speak to her in a low tone, none threateing manner. Recommend a follow up with her doctor or even a new doctor; her band may need to be adjusted, deflated or removed.
These answers are for educational purposes and should not be relied upon as a substitute for medical advice you may receive from your physician. If you have a medical emergency, please call 911. These answers do not constitute or initiate a patient/doctor relationship.


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The cottage has a number of green credentials including; low volt lighting, A+ class fridge/freezer, solar water heater, reclaimed wood beams, stone used on its renovation were supplied from its own land, use of well water, linen is changed once a week in order to save water.
Electricity is supplied from natural resources and runs "Off the National Grid" and gives guests the opportunity to experience firsthand, living with solar panels and wind turbines and using the latest renewable energy systems. It has been installed the most up to date energy systems on offer, and they work very efficiently. The system in is the largest on Lanzarote and runs on a 48 volt system and has no problem powering the 12 dwellings on site. Other green credentials include use of grey water, LED lighting, solar pool heating, use of well water for pool, A+ class fridge, solar water heater, organic fruit and veg and within walking/cycling distance to amenities.
Other environmental practices we follow:
- Waste reduction - We encourage recycling and have bins locally. Any organic waste can be given to our chickens.
- Conserving water - We ask client to be use water sparingly. We use our own well water which is used for the swimming pool, we also use our grey water on our palm trees.
- Conserving electricity We use only Wind turbines and solar panels
- Encouraging wildlife - We have planted many native trees and bushes to encourage birds.
- Organic gardening practices - The retreat has a number of fruit trees and plants such as tomatoes, potatoes, spinach, basil strawberries etc..
- Building materials - the entire retreat has been built using stone from the 30,000 meter estate.
- The well and water tanks which are on the land and originally used for irrigating the farm land have been fully restored.
- Measures to educate staff & guests about our environmental practices - We educate guests/staff and local schools on the Eco energy system provided.
- Office practices - We have low energy lighting, and reuse paper.
Local stone and trades men have helped bring the retreat to a beautifully finished paradise. We offer excursions for local schools to visit the retreat and see the effectiveness of the solar/wind system.
We promote rural tourism by encouraging guests to eat out locally and use local amenities such as shops, sports facilities etc. We also encourage them to buy local products. We provided full information on local restaurants, activities and farmers markets.
2 bed villa, shared pool, airport pick-up included
From 700 - 1085 per accommodation per week (sleeps 4)
Luxury large yurt, inc Hybrid car & airport transfers
From 2100 - 2730 per accommodation per week
360º views, ideal couples/single travelers, pick-up incl
From 665 - 910 per couple per week
Beach 100mt, private plunge pool, transfers, Wifi
From 945 - 1365 per accommodation per week