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When dragonflies mate, their tails form a heart.
As we lead up to the QR Code Hunt next week in New York City where you will have a chance to win a brand new Ford Fiesta, I wanted to write some more about how the contest will work and how QR codes w
Deep under the streets of the New York are miles and miles of abandoned subway tunnels.
With focus on the use of the language, what specific trait do the following three three entries have in common?
gray airplane
favorite neighbor
colorful jewelry
Here is the SOLUTION.
What Everyone Needs to Know About College and Privacy.
While the origins of Festivus are a little murky — Seinfeld? The father of a Seinfeld writer? The ancient Romans?
On December 12, 1531, the Virgin Mary, the story goes, appeared on the cloak of a peasant named Juan Diego. Today, the image that appeared on the cloak is popularly known as the Virgin of Guadalupe.
Dan Lewis runs the popular daily newsletter Now I Know ("Learn Something New Every Day, By Email"). We've invited him to share some of his stories on mental_floss this week.
Last week we announced that we're giving away a BRAND NEW FORD FIESTA in NYC through a special QR code hunt a week from today. But what on earth is a QR code?
I enjoy learning about cities that got creative with their official symbols. For example, the plastic pink flamingo is the official bird of Madison, WI.
We're loading up the RV and taking the Mental Floss Trivia Show on the road to beautiful Durham, North Carolina! It's true. This Monday, Feb.
Each year since 2001, Google releases its Zeitgeist summary, showing various lists of top search terms.
Calendars are a popular Christmas gift for friends and relatives who already "have everything", because you need a different calendar every year. And these are certainly different!
Bookstores come in all shapes and sizes, but these 10 featured by WebEcoist are some of the world's coolest locations for selling books. All ten places featured are repurposed buildings and tran
In today's Brain Game, the three definitions that follow apply to three different 8-letter words that are anagrams of one another (meaning they use the same letters of the alphabet).


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Coeur d'Alene (CDE +3.3%) reports lower-than-expected Q2 EPS of $0.31 but a 24% Y/Y gain in revenues to $254M. CDE says it expects to achieve the high end of its 2012 silver and gold production guidance of 18.5M-20M silver oz. and 210K-230K gold oz., and anticipates the low end of guidance for cash operating costs per silver oz. of $6.50-$7.50.
Coeur d'Alene (CDE +3.3%) reports lower-than-expected Q2 EPS of $0.31 but a 24% Y/Y gain in...
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Edgar Bergen
- Birthday
- Feb 16, 1903
Edgar Bergen was still in grammar school when he sent away for a 25-cent ventriloquism instruction book. By the time he was 11, Bergen was driving his family crazy with his prankish voice-throwing. While attending medical school at Northwestern University, Bergen paid his tuition by performing a small-time ventriloquist act; it wasn't long before he… More Bio:
Edgar Bergen was still in grammar school when he sent away for a 25-cent ventriloquism instruction book. By the time he was 11, Bergen was driving his family crazy with his prankish voice-throwing. While attending medical school at Northwestern University, Bergen paid his tuition by performing a small-time ventriloquist act; it wasn't long before he dropped out of college to hit the vaudeville and tent-show circuit. After a tour of Europe and South America, Bergen filmed a series of one-reel short subjects for Vitaphone between 1930 and 1935; even in these early efforts, top billing went not to Bergen but to his creation, the impish, top-hatted dummy Charlie McCarthy. From time-to-time Bergen would test out other wooden alter egos, including hayseed Mortimer Snerd and man-hungry Effie Clinker, but Charlie would remain his star attraction. After gaining nationwide fame through his appearances on Rudy Vallee's radio program, Bergen launched his own radio series, The Charlie McCarthy Show, a top-rated endeavor which ran from 1937 through 1955. Bergen and Charlie made their feature film debuts in The Goldwyn Follies (1938). They went on to appear together in eight more films between in 1938 and 1948; curiously, however, Charlie McCarthy came across better on radio than he did on screen--partly due to the inescapable fact that Bergen tended to move his lips while throwing his voice. At his peak, Bergen was pulling down $10,000 weekly from his radio series and an additional $100,000 from Charlie McCarthy toys and merchandise. He was also the recipient of the only wooden Academy Award in history, a special Oscar bestowed upon himself and the pine-headed Charlie. On his own, Bergen co-starred in I Remember Mama (1948) as the shy Norwegian suitor of spinster Ellen Corby; he also played supporting parts in Captain China (1949) and Don't Make Waves (1965). After the cancellation of their radio series, Bergen and Charlie played nightclubs, summer stock and state fairs. They also hosted the 1956 TV quiz show Do You Trust Your Wife?. Emerging from a long professional slump, Bergen made a triumphant Las Vegas comeback in 1978--the evening before his death at the age of 75. His last screen appearance was in Jim Henson's The Muppet Movie (1979), which was dedicated to his memory. Edgar Bergen was the husband of actress Frances Bergen, and the father of film and TV star Candice Bergen.
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;woohoo! We've reached chapter 10! Please note: this whole chapter is a flash back to Lucky's past. As always I don't own captain America just half the plot line and any OC's;
The streets of Brooklyn where busy as ever on the hot summer afternoon. Business men and Families wandered through the streets trying to get home or to where they needed to be.
A little trio walked through the crowded sidewalks, hoping to get to the park before they had to return home.
One boy was rather scrawny and small for an eight-year-old, with blonde hair and ocean blue eyes was talking to his much taller, more built brown haired friend.
They talked about soldiers, guns and other military weapons while a little girl desperately tried to keep up, clinging the taller boys hand. Her crystal blue eyes looked annoyed.
"Bucky! Stop walking so fast! I can't keep up!" She whined jogging to catch up to the boys.
"C'mon, Luck, you can keep up." Bucky said tugging her hand to pull her forward. "You don't make it easy..." Lucky huffed as she threw one of her pig tailed braids over her shoulder.
The three children finally made their way to park where several other children where already playing.
"Hey Bucky!" A random red headed boy ran up holding a foot ball in his hands. "Want to play?" He asked indicating the large mass of boys that where Bucky's age.
"Can I play?" Lucky asked. The boy looked at her like she was plain stupid. "No way! Your a 6 year old girl." He retorted.
Lucky scowled at him. "I don't see why she shouldn't." Steve cut in. The red head smirked, "She would cry the moment one of them pushed her." He said. "And you," he said indicating Steve. "You'd be broken like a- OW!"
Lucky had ran up and kicked the red head in the shin. Before he could recover, she ran next to Bucky. The red head scowled and advanced towards her.
Bucky stepped in between them, blocking his little sister from view. He was a lot taller than the kid, and far more stronger. "I don't think I want to play today, Lenny." He said coolly while taking Lucky's hand.
The six year old looked at Lenny from behind Bucky and stuck her tongue out at him as he leaded them away.
"I think you over reacted I bit there, Luck." Steve said looking back at the kids.
Lucky scowled. "He said I couldn't play cause I'm a girl! And he said you'd snap like a twig." She added.
"Technically, you kicked him before he could finish his sentence." Bucky corrected her. "Still, I wasn't going to say "yes" any way."
"It was still fun." Lucky mumbled as they made their way to the swing set.
Few hours later, the sun was going down and the trio was ready to go home.
"C'mon, Luck." Bucky said extending his hand out to help her stand up out of the grass. "Bucky I'm tiered." Lucky yawned as she rubbed her eyes.
Bucky sighed and picked lucky up onto his back and gave her a piggy-back-ride home.
"Night, Steve." Bucky called to the small blonde as he made his way up the front steps. "Night." Lucky called. Steve waved goodnight and headed inside.
Bucky carried Lucky a block down to reach their own home. "C'mon, Luck, I can't open the door and carry you." He said setting her down.
"Mom! We're home!" Bucky called leading Lucky inside. "Hey! There's my favorite kids!" A
tall man with dark brown hair and brown eyes walked into the hallway wearing a military uniform grinning ear to ear.
"Daddy!" Lucky squealed running into her fathers arms. "Hey, Pumpkin." He said picking her up. Lucky pouted. "Daddy, my name is "lucky"! Only lucky!" She said crossing her arms.
"Only lucky, eh?" He asked. "I can't even call you "pumpkin"?" Lucky shook her head. ""Sweat heart"? "Darling"? Or even the name your mother and I gave you?" Lucky shook her head. "No. No. No. And no. Only "Lucky"."
He laughed as he hugged Bucky with one arm. "How us doing champ? Take care of your mother and sister while I was away?"
Bucky nodded. "It'd be easier if lucky wasn't running around and kicking people in the shin." "That's because Lenny wouldn't let me play football with them!" Lucky added defensively.
Their dad just laughed and kissed Lucky on the cheek. "That's my girl!" He laughed.
With Lucky in one arm, and the other on Bucky's shoulder, he lead them to the kitchen.
;thanks for reading please review!;


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As a PTA you'll work under the direction and supervision of the physical therapist and help to carry out the physical therapy treatment plan. Watch the You Can Be Me video!
To be successful in this career path, you should truly enjoy working closely with people and studying science courses, such as anatomy and physiology.
You can access further information about careers in physical therapy at: American Physical Therapy Association.
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Effective 7/1/15, all students testing at NVCC will be charged a proctor fee (payable to NVCC) in addition to ATI's registration fee. The proctor fee must be received by the Cashier's Office prior to the day of the test. Students who have not paid the proctor fee will not be permitted to take the TEAS.
Students must score a minimum 50% on the TEAS to be eligible for admission to the PTA Program.
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Show transcribed image text Design a lowpass Chebyshev filter with the following characteristics: Amax=0.1dB A min=30dB fp=10kHz fx=15kHz Provide a worksheet showing your calculations for the following variables (write the answer in here): If you were to design this filter, how many stages would you need? Describe this filter in terms of first-order and second-order filter stages: Use Excel to plot the filter response (using the Chebyshev expression for the filter magnitude.) Show A max, A min, omega p, and omega s on your graph Do not draw these in by hand Use Excel functionality to add lettering and pointers on your graph Hand this Excel graph in with your worksheet showing all your calculations (see "What to bring to lab" at the end of this document.) Is this a practical filter to fabricate? Why or why not? How does this compare to a Butter worth filter of the same order? What would be the value of A min?


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Watch Boston's WCVB TV Channel 5 reporter Kelley Tuthill, a breast cancer survivor treated at Dana-Farber, share her tips for selecting a wig and wearing it with confidence. Learn how to wear a wig, keep it in place, find the right look, and more.
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Vasopressin acts on the inner medullary collecting duct (IMCD) in the kidney to regulate water and urea transport. To obtain a 'parts list' of gene products expressed in the IMCD, we carried out mRNA profiling of freshly isolated rat IMCD cells using Affymetrix Rat 230 2.0 microarrays with approximately 31,000 features. 7913 annotated transcripts were found to be expressed above background in the IMCD cells. We have created a new online database (the "IMCD Transcriptome Database" †) to make the results publicly accessible. Among the 30 transcripts with the greatest signals on the arrays were three water channels: aquaporin-2, aquaporin-3, and aquaporin-4, all of which have been reported to be targets for regulation by vasopressin. In addition, the transcript with the greatest signal among members of the solute carrier (SLC) family of genes was the UT-A urea transporter (Slc14a2), which is also regulated by vasopressin. The V2 vasopressin receptor was strongly expressed, but the V1a and V1b vasopressin receptors did not produce signals above background. Among the 200 protein kinases expressed, the serum-glucocorticoid regulated kinase (Sgk1) had the greatest signal intensity in the IMCD. WNK1 and WNK4 were also expressed in the IMCD with a relatively high signal intensity as was protein kinase A (β-catalytic subunit). In addition, a large number of transcripts corresponding to AKAPs and 14-3-3 proteins (phospho-S/T binding proteins) were expressed. Altogether, the results combine with proteomics studies of the IMCD to provide a framework for modeling complex interaction networks responsible for vasopressin action in collecting duct cells.
The inner medullary collecting duct (IMCD) is the terminal portion of the collecting duct system of the kidney. The collecting duct system represents the final site of adjustment of urinary composition and volume, and therefore is critical for extracellular fluid homeostasis. An important regulator of collecting duct transport function is vasopressin, which controls both water and urea transport (21; 53)
To address the mechanisms of vasopressin signaling in the renal IMCD, we have been following a "systems–biology" approach consisting of identification of the component proteins via protein mass spectrometry-based analysis (2; 29; 30; 62; 71; 102), and antibody-based quantification of protein abundance (41), coupled to computational analysis of the identified proteins to discover signaling networks involved in IMCD regulation (30). An important product of these studies is the generation of a 'parts list', the IMCD proteome, that includes the signaling apparatus (http://dir.nhlbi.nih.gov/labs/lkem/rm/proteomics_db.asp). The IMCD proteome database currently is made up of 2338 proteins identified at high stringency.
Despite the success so far of the proteomics approach, proteomics methodologies have limitations that make it unlikely that all of the proteins that play important roles in vasopressin signaling will be found. Limited sensitivity is the chief factor. In the present study, in order to augment our enumeration of an 'IMCD parts list', we have pursued an analysis of the IMCD transcriptome using Affymetrix oligonucleotide arrays. The studies were carried out in native IMCD cells freshly isolated from rat inner medullas. The arrays contain approximately 31,000 features representing most of the expressed genes in the rat genome. Although the approach is comprehensive, the chief objective is to determine what transcripts are expressed in functional categories most relevant to vasopressin signaling related to short- and long-term regulation of water and urea transport in the IMCD.
The new data from the present study add to transcriptomic data obtained in microdissected cortical and outer medullary collecting ducts using a micro-SAGE technique (9). SAGE data, however, are not available for the IMCD. Early approaches to transcriptional profiling in IMCD involved large-scale sequencing of cDNAs prepared from microdissected IMCD segments, an approach that was informative but not comprehensive (78). A number of studies have also reported transcriptional profiling of whole inner medulla (7; 8; 51; 66; 80), which contain IMCDs as well as loops of Henle, vasa recta, capillaries and interstitial cells. IMCDs make up about 40% of the inner medullary tissue by volume (40). In the present study, we carry out transcriptional analysis of biochemically enriched IMCD cells isolated from rat renal inner medulla.
Pathogen-free male Sprague-Dawley rats weighing 100–120 grams (Taconic Farms Inc. Germantown, NY) were maintained on an autoclaved pelleted rodent diet and ad libitum drinking water (NHLBI Animal Care and Use Committee approved protocol number H-0110). The age of the rats was chosen to match the age of rats typically used for our isolated perfused tubule studies (12).
IMCD and non-IMCD cells were prepared from whole inner medullas as previously described (62) with slight modification (Figure 1 and Supplementary Methods).
In order to assess purity of IMCD samples, aliquots from each cell suspension were loaded onto SDS-polyacrylamide gels and immunoblotting was carried out as described (12). An immunoblot for a collecting duct marker AQP2 (Figure 2 panel A) demonstrated that AQP2 was indeed highly enriched in the IMCD fraction and depleted from the non-IMCD fraction (Band densities: IMCD fraction, 412; non-IMCD fraction, 45). An immunoblot for AQP1, a marker of vasa recta and thin descending limbs of Henle (Figure 2 panel B), demonstrated that there was relatively little contamination of the IMCD fraction from non-IMCD elements (Band densities: IMCD fraction, 1.4; non-IMCD fraction, 29.4). Lack of contamination by outer medullary cells was demonstrated by immunoblotting using an antibody to NKCC2, a marker of the outer medullary thick ascending limb (Figure 2 panel C) (Band densities: IMCD fraction,0.9; non-IMCD fraction, 0.4; OMCD fraction, 11.3).
Total RNA of IMCD and non-IMCD cells was extracted using Trizol reagent (Invitrogen, Carlsbad, CA) following the manufacturer's protocol. Full details are reported in the Supplementary Methods. Typically, for IMCD, the yield was 1 μg per kidney and, for non-IMCD, the yield was 0.7 μg per kidney.
Hybridization probes (labeled cDNAs corresponding to the transcripts) were prepared and hybridized to the Affymetrix Rat Genome 230 2.0 microarrays (Affymetrix Inc, Santa Clara, CA) according to the manufacturer's protocols. See Supplementary Methods for full details.
The total number of genes with the Affymetrix Detection p-value consistent with a "Present Call" (detected) was about 17,000 from 31,000 features in the Affymetrix Rat Genome 230 2.0 array for both IMCD and non-IMCD samples. These 17,000 features corresponded to about 10,200 distinct genes. The remaining 14,000 features were identified to have "Absent" (not detected) or "Marginal" Calls based on the Detection p-value from Affymetrix GCOS software. Intensities of each probe set were normalized by median-transformed normalization.
The IMCD and non-IMCD samples contain a relatively small amount of contaminating cells from non-IMCD elements and IMCD elements, respectively (see AQP1 and AQP2 immunoblots in Figure 2). We calculated corrected values for mRNA signals using estimates of cross contamination. The appropriate equations are (see Supplementary Methods for derivation):
IMCDcorrected = (d·IMCDobserved — b·non–IMCDobserved)/(ad–bc)(Equation 1)
non–IMCDcorrected = (a·non–IMCDobserved — c·IMCDobserved)/(ad–bc)(Equation 2)
IMCDobserved = the normalized signal intensity for a given feature for IMCD sample;
non–IMCDobserved = the normalized signal intensity for the same feature for the corresponding non–IMCD sample;
a = fraction of the total signal in IMCD samples from IMCD cells
b = fraction of the total signal in IMCD samples from non–IMCD cells
c = fraction of the total signal in non–IMCD samples from IMCD cells
d = fraction of the total signal in non–IMCD samples from non–IMCD cells.
Note also that a + b = 1 and c + d = 1.
The cross contamination values a, b, c, and d could in principle be estimated from the AQP1 and AQP2 immunoblots but a more general approach is to use the array data to determine the required cross contamination correction. Supplementary Figure 3 shows a histogram of the ratios of normalized, but uncorrected, IMCD-to-non-IMCD signal ratios for individual features on the array. In theory, the maximum and minimum ratios should be bounded at a distinct value determined by the degree of cross contamination. The maximum and minimum ratios in principle could then be used to estimate the degree of cross contamination. However, as seen in Supplementary Figure 3, there is considerable splay in the tails of the ratio histogram, indicating that a direct choice of single unique values for cross contamination is not feasible. Consequently, we chose an alternative approach in which we chose a set of transcripts corresponding to IMCD and non-IMCD marker proteins. These are transcripts known to be exclusively or predominantly expressed in the IMCD ("IMCD markers) and transcripts known to be expressed in non-IMCD structures but not in the IMCD itself ("non-IMCD markers"). Thus, we choose values for a, b, c, and d such that the mean value for [non-IMCDcorrected] is zero for a selected set of IMCD markers and the mean value for [IMCDcorrected] is zero for a selected set of non-IMCD markers. For IMCD markers, we have chosen aquaporin-3 (Aqp3), gamma ENaC (Scnn1g), ClC-K2 (Clcnkb), the vasopressin V2 receptor (Avpr2) and frizzled receptor 1 (Fzd1). For non-IMCD markers, we have chosen UT-B (Slc14a1, expressed in vasa recta), NHE3 (Slc9a3, expressed in thin descending limb of Henle), von Willebrand's factor (Vwf, expressed in endothelial cells of vasa recta) and EEAC1 (Slc1a1, expressed in thin limbs of Henle's loop). Calculated in this way, the resulting values for b, c, and d were: a = 0.80 ± 0.00, b = 0.20 ± 0.00, c = 0.27 ± 0.01 and d = 0.73 ± 0.01.
Equations 1 and 2 were applied to each feature to gain corrected signal intensities that would be expected if the samples were pure IMCD and non-IMCD suspensions. The signal intensities for IMCD and non-IMCD are reported as mean ± SE for all replicates.
The general strategy employed in this study was to use Affymetrix microarrays to profile gene expression at an mRNA level in rat IMCD cells, focusing on the general question: What gene products can be identified in the IMCD that could be components of the pathways and networks involved in regulation of aquaporin-2 and osmotic water permeability in the IMCD? The data presented are calculated from the results of 6 microarray chip sets, i.e. 3 pairs of IMCD vs. non-IMCD transcripts. Total RNA samples for each pair were isolated from 7 rats (14 inner medullas). A full listing of the results is shown in Supplementary Table S1 and the full results have also been deposited in the GEO database (GSE7891: at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=rbyjrsiskikmeze&acc=GSE7891). 7913 annotated transcripts (compared to 10,206 total transcripts above background) were found to be expressed above background in the IMCD cells.
We have created a new online database (the "IMCD Transcriptome Database" at http://dir.nhlbi.nih.gov/papers/lkem/imcdtr/) to provide ready access to these data. This site also includes the data in a downloadable format and a BLAST facility for searching for a particular transcript in the database.
The variability of the normalized, corrected signals for the three experiments is shown as scattergrams in Supplementary Figure 4. As can be seen, the distribution of IMCD and non-IMCD signals were reproducible with more than 85% of IMCD features and more than 80% of non-IMCD features showing coefficients of variation (CV = variance/mean ×100%) less than 15%. Supplementary Figure 5 shows mean corrected, normalized signals for IMCD plotted against the non-IMCD values for the same features. Values that deviate from the diagonal indicate transcripts selectively expressed in IMCD (above line) or non-IMCD elements of inner medulla (below line). Supplementary Figure 6 shows the corrected values for IMCD plotted against the corresponding non-corrected values. As can be seen, the corrections were generally relatively small. Only 0.8 percent of the values had correction factors of 50% or more of the uncorrected values.
In the following, we present a series of tables showing, for specific categories of gene products, what transcripts in IMCD cells are associated with the highest signals on the microarrays along with comments regarding gene products that are thought to be involved in vasopressin action in the IMCD. We also include significant negatives in which corrected signals in the IMCD were indistinguishable from zero. The classification of expressed genes in each table was based on classification terms from the PANTHER Database (83) and the Rat Genome Database (87). We emphasize that the signals from the array provide a general index of relative expression levels for individual transcripts but do not necessarily correlate with the absolute level of the specific mRNA in the cells. Furthermore, transcript level does not necessarily correlate with protein expression level owing to differences in translation rate and protein stability among various proteins (7).
Table 1 summarizes the microarray results for G-protein coupled receptors. Normalized and corrected intensity values reported in all tables are calculated from the raw data as described in the Methods section. The value of 1.0 represents the median value for all positive transcripts in the IMCD. A number of receptors with well established roles in the IMCD were identified and had substantial signals on the arrays. These were the vasopressin V2 receptor, the frizzled receptor 1, the PGE2 receptor EP1, the endothelin B receptor, the adenosine A1 receptor, the adrenergic receptor 2a, the frizzled receptor 6, the adrenergic receptor 1d, the PGE2 receptor EP4, and the nucleotide receptor P2Y2. Of these, only the vasopressin V2 receptor (53) and the prostaglandin E2 receptor EP4 (5) are known to be coupled to the heterotrimeric G-protein Gs and to generate cyclic AMP when bound to ligand. Presumably, PGE2 does not increase water permeability in the IMCD because of the prevailing inhibitory effect of the EP1 receptor, which signals via Gαq. In addition to receptors with well-established roles, a number of other G-protein coupled receptors were identified in the IMCD (Table 1), providing hypotheses for further study. For example, the chemokine orphan receptor 1 (Cmkor1) is the highest ranked receptor from the point of view of normalized intensity.
In our previous study, vasopressin-induced cAMP production was inhibited by epinephrine or the α2-agonist clonidine, but not by the α1-agonist, phenylephrine (46). This finding is consistent with the present finding of dominant expression of the α2a-adrenergic receptor relative to the α1-adrenergic receptor as shown in Table 1. In contrast, β-adrenergic receptor expression was below background.
The adenosine receptor A1 is associated with a high signal intensity on the IMCD arrays (Table 1). It has been shown that adenosine analogues can decrease water permeability induced by arginine vasopressin (18). Our previous experiments using the adenosine A1 agonist NPEA (N6-2-phenylethyladenosine) showed an increase in intracellular Ca2+ concentration and this effect was inhibited by the selective adenosine A1 antagonist 8-phenyltheophylline (17).
Endothelin-1 can induce diuresis in part by inhibiting water reabsorption induced by arginine vasopressin in the IMCD (60). In the IMCD, it has been demonstrated by immunofluorescence and RT-PCR that the endothelin B receptor is the predominant endothelin receptor subtype in agreement with our microarray data (42; 81).
Purinergic P2Y2 receptors have been demonstrated in the rat IMCD (38). Binding of ATP or UTP to P2Y2 receptors increase intracellular calcium in the IMCD, decreasing water permeability (17). This inhibition of water permeability might occur through activation of protein kinase C and reduction of cellular cAMP levels (37).
Arginine vasopressin receptors are of two types, V1 and V2 receptors. From the microarray analysis, the signal intensity corresponding to the V1a and V1b receptors in the IMCD cells was negligible. In contrast, V2 receptor signal intensity was relatively high. This evidence supports our previous finding that V2 receptors, but not V1a receptors, are expressed in the IMCD (16; 45).
Table 2 shows heterotrimeric G-protein subunits expressed in the IMCD. The α subunits expressed include three inhibitory α subunits (Gαi) which regulate adenylyl cyclases in an inhibitory fashion, the stimulatory α subunit (Gαs) which stimulates adenylyl cyclases, Gαq which couples to phospholipase Cβ (PLCβ) isoforms and Gα12, an activator of Rho-GEF (43) and a regulator of the actin cytoskeleton. All are potentially involved in vasopressin signaling.
Only 3 adenylyl cyclase transcripts of the 10 known genes were identified in the IMCD samples, namely Adcy6 and Adcy3 previously identified in the IMCD (28), as well as Adcy9 as shown in Table 3. In addition, soluble guanylyl cyclase 1 was identified, which functions as a nitric oxide receptor and transducer. Interestingly, there was no significant signal for any of the three nitric oxide synthase (NOS) isoforms in the IMCD samples, although NOS3 (endothelial NOS) was detected in the non-IMCD samples (Table 16). However, previous qualitative RT-PCR studies in microdissected IMCD segments demonstrated the presence of NOS1, 2 and 3 mRNA (96). Furthermore, both NOS1 (102) and NOS2 (91) protein has been detected in the IMCD by protein mass spectrometry.
As shown in Table 4, the predominant cyclic nucleotide phosphodiesterase transcripts correspond to the type 4 and type 1 classes, consistent with pre-existing findings (15; 75). Type 4 phosphodiesterases (PDE4A, PDE4B, PDE4C, and PDE4D, coded by separate genes) are cAMP–specific and inhibitable by rolipram. Type 1 phosphodiesterases hydrolyze both cAMP and cGMP and are calmodulin-dependent. No significant signal was found for PDE5, which has been implicated in regulation of aquaporin-2 trafficking in more proximal collecting duct segments (4).
There are more than 500 genes coding for protein kinases in mammalian genomes (48). As shown in Table 5, a large number of these are expressed in the IMCD. There were 93 serine/threonine kinases with signals above the median value in the IMCD (corrected signal above 1.0, Table 5A). Serum-glucocorticoid regulated kinase, sgk1, a serine/threonine kinase closely related to protein kinase B (Akt1), is the kinase with greatest signal intensity in the IMCD (Table 5A). This kinase is strongly regulated by the mineralocorticoid hormone aldosterone (11). WNK1 and WNK4, two kinases implicated in regulation of ion transport in more proximal parts of the renal tubule (95; 99), are also expressed in the IMCD with a relatively high signal intensity. Protein kinase A (catalytic β subunit), long recognized as a major element of vasopressin signaling, is also associated with a high signal intensity on the IMCD arrays. Additional protein kinases with proposed roles in vasopressin signaling have relatively high signal intensities including protein kinase B (Akt1), protein kinase C (iota, zeta, nu and epsilon), casein kinase isoforms, calmodulin-dependent kinases, myosin light chain kinase, Rho-kinase, two G-protein coupled receptor kinases (GRK2 and GRK6), various kinases in MAP kinase pathways, and cyclic-GMP-dependent kinases. Obviously, a large number of kinases were detected whose roles in the IMCD have not been investigated.
Relative to the number of serine/threonine kinases in IMCD, There are relatively few tyrosine kinases (Table 5B and 5C). Tyrosine kinases are of two types: non-receptor tyrosine kinases and receptor tyrosine kinases. Of the non-receptor tyrosine kinases, Frk (a src-family homolog) is strongly expressed in IMCD relative to non-IMCD elements. Also strongly represented is Janus kinase 1 (Jak1), which regulates transcription through activation of Stat1 and/or Stat2. Of the receptor tyrosine kinases, two are heavily expressed in the IMCD relative to non-IMCD elements, namely fibroblast growth factor receptor 2 (Fgfr2) and epidermal growth factor receptor (Egfr). Most of the detectable tyrosine kinases in the IMCD have been identified previously in fetal kidney including Jak1, Frk, Ptk2, Fgfr2, Egfr, Ryk, Ddr1 and Fgfr1 (36). Tyrosine kinases are associated with regulation of cell proliferation and differentiation.
The serine/threonine phosphatases found in the IMCD are listed in Table 6A. Focusing on the catalytic subunits, the highest signal intensities in the IMCD appear to be the PP1 phosphatases of which three different isoforms are represented namely, α, β and γ. The catalytic subunit of calcineurin, a Ca2+-calmodulin dependent phosphatase (PPP3CA), is also expressed as is the catalytic subunit of the magnesium-dependent phosphatase, PP2C. For all of these catalytic subunits, significant expression was found in both the IMCD and non-IMCD fractions, consistent with their ubiquitous roles in cell signaling (69). In contrast, the PP2A catalytic subunit signal on the IMCD arrays was indistinguishable from background, although several of its regulatory subunits were present. Calcineurin has been reported to be expressed in the IMCD and to be colocalized with PKCζ, the RII subunit of PKA, and AQP2 in endosomes of the IMCD (35). It has been reported that the α isoform of the calcineurin catalytic subunit is required for normal intracellular trafficking of AQP2 (23). Finally, there are many more genes in the genome that code for Ser/Thr phosphatase regulatory subunits than for catalytic subunits, a number of which were found to be expressed in the IMCD.
Table 6B summarizes the protein tyrosine phosphatases (PTPs) found in the IMCD. The protein tyrosine phosphatases can be classified into transmembrane receptor-like protein tyrosine phosphatases and cytosolic protein tyrosine phosphatases (84). There are 16 PTP transcripts identified in the IMCD samples with signal intensity above median. Ten coded for cytosolic PTPs and 5 for transmembrane receptor-like PTPs.
Dual specificity phosphatases in the IMCD are summarized in Table 6C. These phosphatases dephosphorylate a phosphotyrosine and a neighboring phosphothreonine. Among these, the transcript corresponding to Dusp1 displays the strongest intensity in the IMCD. Its function is to dephosphorylate the MAP kinases ERK1 and ERK2 at the sites phosphorylated by dual specificity kinases (MAP kinase kinases or MEKs) (73). Two dual specificity phosphatases are expressed in the IMCD but not in non-IMCD cell types, namely Dusp2 and Dusp5.
A-kinase anchoring proteins (AKAPs) are scaffold proteins that facilitate localized signaling through creation of binding complexes that bring enzymes (mainly kinases and phosphatases) into the vicinity of their targets. Table 7 shows the AKAPs whose signal intensity in the IMCD samples were above background. Those expressed at highest levels were AKAP-7, -8, -9, -11 and -13. AKAP7 (aka AKAP18δ) has been shown to be colocalized with AQP2 intracellularly in the IMCD (27). Both AKAP7 and AQP2 were localized predominantly at the apical plasma membrane when the IMCD cells were stimulated with dDAVP.
14-3-3 proteins are phospho-serine/phospho-threonine binding proteins. By virtue of their binding specificities, they have been proposed to play various phosphorylation-dependent regulatory roles. There are seven isoforms of 14-3-3, viz. β, γ, ε, η, σ, τ and ζ (Note: α and δ are phosphorylated forms of β and ζ isoforms (97; 98)). In the present study, 5 of 7 isoforms of 14-3-3 were identified in the IMCD with high signal intensity (Table 15). Functional roles of 14-3-3 in the IMCD have not been elucidated yet. However, 14-3-3 proteins have been found to modulate the function of the ubiquitin ligase Nedd4-2 in cultured collecting duct cells by binding to Nedd4-2 after sgk1-mediated phosphorylation (3).
Table 8 shows the phospholipases in IMCD with signals above background for either IMCD or non-IMCD samples. Among the phospholipases with the highest signal intensities was cytosolic phospholipase A2 (PLA2) which hydrolyzes the ester bond in SN-2 position of the core glycerol. This reaction is important because it liberates arachidonic acid, an unsaturated fatty acid that is the substrate for prostaglandin synthesis. Vasopressin has been seen to be variably associated with stimulation of prostaglandin E2 synthesis in collecting ducts, which provides a modulating influence on water permeability (68). Prostaglandin E2 synthesis depends on expression of one of two cyclooxygenases (COX-1 and COX-2). COX-1 exhibited a very high signal on the arrays, with a signal that is in the top 100 among transcripts in the IMCD. Both COX-1 and COX-2 are selectively expressed in the IMCD. As noted above, three different PGE2 receptors are expressed in the IMCD consistent with the view that PGE2 works as an autocoid.
As shown in Table 8, several isoforms of phospholipase C (PLC) are also expressed in the IMCD. PLC cleaves phosphatidylinositol 4,5-bisphosphate (PIP2) to yield two second messengers, diacylglycerol and inositol 1,4,5 tris-phosphate (IP3). The beta isoforms are activated by heterotrimeric G-protein alpha subunits Gαq and Gα11. There are two PLC-β isoforms expressed in the IMCD, PLCβ1 and PLCβ4. PLCβ1 is expressed selectively in IMCD cells versus non-IMCD cells. PLCγ isoforms are regulated via tyrosine phosphorylation, typically following growth factor receptor activation (65). PLCγ1, but not PLCγ2, appears to be expressed in IMCD substantially above ackground. PLCδ4 is also selectively expressed in the IMCD over non-IMCD cell types.
Table 9 lists transcripts corresponding to small GTP-binding proteins expressed on the IMCD arrays including Rab proteins, Arf/Sar proteins, Rho/Rac/CDC42 proteins, Ras and Ras-like proteins, and Ran proteins (77).
Members of the Rab family of proteins play a critical role in intracellular membrane trafficking. Large numbers of transcripts corresponding to Rab proteins were identified on the IMCD expression arrays (Table 9A) including many of those previously identified in a proteomic analysis of aquaporin-2-containing vesicles in IMCD cells (2). Most of the Rab proteins were found both in IMCD and non-IMCD cells, although Rab15 and Rab31 appear to be selectively expressed in the IMCD. Based on studies in non-renal cells, Rab15 is believed to be expressed in early endosomes and recycling endosomes (74). Rab31 is thought to be an early endosomal Rab based on its similarity to Rab5 (72).
ADP-ribosylation factors (Arfs) regulate coat protein-mediated vesicle budding, vesicular trafficking, and actin cytoskeleton assembly. Transcripts corresponding to most of the known Arf and Arl (ADP-ribosylation factor-like) proteins are expressed in the IMCD (Table 9B). Arf1 has been shown to be important for Golgi to ER transport. Arf6 is important for endocytosis (10; 14). Both Arf1 and Arf6 displayed high signal intensity on the arrays. Transcripts for the Arf-like proteins Sar1a and Sar1b were also found in the IMCD.
Rho/Rac/CDC42 GTPases are involved in regulation of the actin cytoskeleton and cell mobility. CDC42, Rnd1, Rac1, and RhoA are the four members of this family with the greatest signals on the arrays (Table 9C). In IMCD cells in primary culture, cAMP was found to inhibit Rho activation, thereby stimulating actin depolymerization and aiding translocation of aquaporin-2 to the plasma membrane (90). Rnd1 is expressed at a very high level in IMCD relative to other inner medullary cell types, specifically about 20-fold higher. Rnd1 has also been reported to promote disassembly of actin filaments (55). However, whether it plays a role in aquaporin-2 regulation has not yet been reported. A number of other Rho-like GTPases were also associated with high signal intensities in the IMCD including RhoB, RhoC, RhoG, RhoQ, RhoT2, and Rnd3.
The Ras-like protein family of small GTPases includes Rap, Rheb and Ral as well as Ras itself. They play an important role in regulating cell proliferation and differentiation. Among the Ras and Ras-like protein transcripts expressed in the IMCD, Rheb was associated with the greatest signal on the arrays (Table 9D). Rheb is a small GTPase that binds and activates the mTOR kinase (1). Another transcript with a relatively high signal intensity was K-Ras. This isoform of Ras has been shown to be an aldosterone-stimulated protein in collecting duct cells (93). Transcripts corresponding to other Ras-like proteins expressed in the IMCD include RalA, RalB, Rap1a, Rap1b, Rasd1, and Rras2. The Ral proteins have been found in aquaporin-2 containing intracellular vesicles in the IMCD (2). These proteins are associated with the so-called 'exocyst complex' involved in basolateral targeting of proteins. We have identified RalA in the basolateral plasma membrane of IMCD cells (unpublished data). Rap1 is involved in cell morphogenesis, cell differentiation, cell adhesion and cytoskeleton organization. It has been demonstrated that SPA-1 (signal-induced proliferation-associated 1), a GTPase-activating protein specific to Rap1, binds to AQP2 in the renal medulla (56). We identified SPA-1 in the IMCD in this study (Supplementary Table S1). Studies in SPA-1 knockout mice showed a defect in AQP2 trafficking to the apical plasma membrane (58). Rap1 is also a target of Epac, a RapGEF, that is activated by cAMP. Epac has been found to be important for regulation of vasopressin-simulated calcium mobilization in collecting duct cells (101), which is necessary for AQP2 trafficking (13). Transcripts corresponding to Epac1 but not Epac2 were identified in the IMCD in our array studies (Supplementary Table S1). One of Rap1's targets is Raf1, a kinase in the MAP kinase pathway (see above). A full listing of transcripts corresponding to small GTP-binding protein GEFs, GAPs and interacting proteins found in the IMCD is reported in (Supplementary Table S4).
SNARE proteins play an important role in membrane fusion in eukaryotic cells. Syntaxin 7 and 12 were the SNARE protein transcripts with the greatest signals in the IMCD as shown in Table 10. Syntaxin 7 is present in the late endosome while syntaxin 12 is present in the recycling endosome (63), an organelle that may be important in AQP2 recycling to the plasma membrane. Plasma membrane syntaxins include syntaxins 2, 3 and 4. Among these, only syntaxin 4 was associated with a strong signal in the IMCD. This syntaxin is present in the apical plasma membrane and has been proposed to be involved in AQP2 trafficking (47). In the current study, VAMP3 (cellubrevin) had the strongest signal intensity among VAMP/synaptobrevin transcripts. This protein was previously identified in the IMCD by immunogold electron microscopy (22). VAMP2 and VAMP4 transcripts were also found to be expressed in the IMCD above the median value. VAMP2 colocalizes with aquaporin-2 in AQP2-containing intracellular vesicles (2; 54). An additional subfamily of SNARE proteins is the SNAP25 homologues. SNAP25 is neural-specific and its expression level was very low in the IMCD. SNAP23 protein has previously been identified in the IMCD on the basis of immunorecognition (32) and in the present study we found that it is expressed in the IMCD above the median expression level for all IMCD transcripts.
Both clathrin heavy chain and light chain polypeptides were identified in the IMCD (Table 11). The clathrin adaptors, AP1, AP2 and AP3 are also expressed in the IMCD. These clathrin adaptor proteins function in post-Golgi trafficking pathways. AP1 regulates vesicle transport between the trans-Golgi network and endosomes. AP2 regulates endocytosis (6) and AP3 is involved in vesicle trafficking from early endosomes to late endosomes or lysosomes (59). In addition, three clathrin adaptors called epsins are strongly expressed in the IMCD namely epsin 4 (enthoprotin), epsin 3 and epsin 2. These proteins bind both to ubiquitylated integral membrane proteins and to PIP2 (in addition to clathrin) and may be involved in regulated endocytosis of apical plasma membrane transporters in the collecting duct (94). Finally, an adaptor called phosphatidylinositol binding clathrin assembly protein or CALM, a PIP2-binding clathrin adaptor was found at a relatively high signal intensity level on the IMCD arrays.
Dynamins are GTPases (although not 'small' GTPases) that are associated with clathrin-coated pits and are involved in the abscission process forming clathrin-coated vesicles. In this study, dynamin-1 was expressed below background and dynamin-2 and -3 were expressed at very low levels only (0.21 and 0.45, respectively, Table 15). In contrast, the dynamin-like GTPases Mx1 and Mx2 were associated with high signal intensities in the IMCD (7.60 and 16.14, respectively). Their locations and roles in the IMCD cell have not been reported.
γ- and β-actin, the two chief forms of non-muscle actin were associated with the highest actin signals on the IMCD arrays. These forms have been seen previously in proteomic studies of the IMCD (2; 62). Vasopressin induces actin filament depolymerization in the IMCD (70), a process that has been shown to be required for AQP2 translocation from intracelluar to apical plasma membrane in cultured CD8 cells (79).
ERM proteins (ezrin, radixin, and moesin) are actin-binding proteins that link actin to the plasma membrane and are involved in signal transduction and cytoskeleton remodeling (85). In the present study, ezrin (Vil2) and radixin (Rdx) were found to be expressed in IMCD at high signal intensity. On the contrary, moesin mRNA was not detected. A previous study (66) reported a marked increased in the expression of ezrin mRNA in the inner medullas of lithium-treated rats, which was associated with an increase in ezrin protein in the IMCD. The authors proposed that this protein plays a role in the nephrogenic diabetes insipidus seen in response to lithium. Moesin has been reported to mediate trafficking of AQP2 to plasma membrane in rabbit CD8 cells (79). Our finding of low or nonexistent levels of moesin mRNA in the IMCD suggests that moesin's role in AQP2 trafficking may not be universal.
Several actin-binding proteins have been identified that are AQP2-binding partners in rat inner medulla including α2-spectrin, α-tropomyosin 5b (tropomyosin 3γ), gelsolin, and α-actinin (57) Transcripts corresponding to all of these proteins showed high signal intensity on our microarrays.
Myosin regulatory light chain B and non-muscle myosin II-B (heavy chain 10) were associated with the greatest signals among myosin gene products. Also associated with relatively high signals in IMCD were myosin II-A (heavy chain 9) and unconventional myosins 1B, 1C, 1D, and 1E, as well as myosin 5B. Myosin regulatory light chain has been demonstrated to be phosphorylated in the IMCD in response to vasopressin (12). This protein regulates the non-muscle myosins II-A and II-B, both of which are expressed in the IMCD. As demonstrated previously, myosin II-B is collecting duct selective in the inner medulla (12). Myosin II-A has been demonstrated to be bound to AQP2 in cell lysates of kidney inner medulla (57). Recently myosin 5B has been reported to be associated with Rab11-positive AQP2-containing endosomes, presumably recycling endosomes (52).
Table 12C shows the transcripts corresponding to the microtubule proteins and microtubule-associated proteins including molecular motors that are expressed above background on the arrays. Microtubule disruption with colchicine and other agents reduces the hydro-osmotic action of vasopressin in collecting ducts (61) through effects on AQP2 trafficking (67). In addition, it has been shown that both dyneins (microtubule-based molecular motors) and dynactin in the inner medulla were associated with trafficking of AQP2-containing vesicles to the apical plasma membrane (49). In the present study, multiple dynein and dynactin isoforms were found in the rat IMCD. Cytoplasmic dyneins are multi-subunit proteins consisting of a combination of light chains, heavy chains, and intermediate chains. The predominant dyneins in the IMCD are cytoplasmic dyneins with signal intensity of light chain-1 > intermediate chain-1 > heavy chain-2 polypeptides. Cytoplasmic dynein heavy polypeptide-2 is selectively expressed in IMCD over non-IMCD elements of the inner medulla.
The transcripts corresponding to transport proteins found in the IMCD above background are reported in Table 13. As can be seen in Table 13A, three water channels are associated with extremely high signal intensities, namely AQP4, AQP2 and AQP3. The signal intensities for these aquaporins were ranked 3rd, 10th and 14th, respectively, among the 7913 annotated transcripts found in the IMCD. In contrast, AQP1 was present only in the non-IMCD samples from the inner medulla.
Table 13B shows the ion channels and transporters expressed in the IMCD. The chloride channel ClC-Kb is expressed in the basolateral plasma membrane of cells throughout the distal nephron from thick ascending limb to the collecting duct and is mutated in some forms of Bartter's Syndrome (92). Its transcript was found at a moderate level in IMCD but not in non-IMCD components of the inner medulla. Note that the levels of the epithelial sodium channel (ENaC) α, β, and γ transcripts are relatively low, consistent with the low level of ENaC protein expressed in this segment (24).
Table 13C shows the non-mitochondrial solute carrier (SLC) transcripts found in the IMCD. Among these, the transcript with the highest signal was the vasopressin-regulated urea transporter UT-A (Slc14a2). This protein is key to the accumulation of urea in the renal inner medulla (19; 20). Its manner of regulation by vasopressin is controversial with one group claiming that the protein undergoes regulated trafficking (39) while another finds an absence of regulated trafficking by vasopressin (33).
Supplementary Table S6 reports the mitochondrial transporters and carriers detected. As may be expected, many of these were components of the F0-F1 ATP synthase responsible for proton gradient-driven ATP synthesis.
We identified about 824 transcription factors expressed in IMCD above background. A list of the transcription factors identified with signal intensity above median is shown in Supplementary Table S7. Only a few of these have been studied in the context of AQP2 regulation.
The 5′ flanking region of AQP2 has been sequenced and putative transcription factor binding sites have been identified including AP1, AP2, CRE, GATA, and Sp1 motifs in human, mouse and rat (64; 88).
Long-term increases in ambient vasopressin in animals or cultured cells results in increased AQP2 protein abundance that is mediated at least in part by enhanced AQP2 gene transcription rates (53). A cAMP responsive element (CRE) plays a critical role (31; 50; 100). It has been widely assumed that regulation of AQP2 transcription involves the transcription factor CREB. However, in this study, we found that CREB itself (now called Creb1) is associated with an extremely low signal intensity on the arrays (Table 14A). This finding raises the possibility that other CREB-family members may be responsible for the long-term regulation of AQP2 gene transcription. The CREB-like transcriptional factors in the IMCD associated with high signal intensities are Atf3 and Atf4 (Creb2). All of these factors share the same recognition site TGACGTCA, which is present in the AQP2 promoter region. Atf3 is thought to be a stress-response transcription factor (25). We hypothesize that Atf4 (Creb2) mediates the effects of cAMP on AQP2 gene expression.
Previous studies have demonstrated that Fos expression is markedly increased in rat inner medulla in response to vasopressin (7). Furthermore, c-Jun phosphorylation was found to be increased in the same study. These two proteins together constitute the transcription factor AP1. They were both found on our arrays with very high signal intensity (Table 14A)
Kruppel-like factors (KLF) are transcription factors in the zinc finger family that regulate cell differentiation, cell proliferation and development with a DNA-binding site motif of CACCC (34; 86). KLF12 was found to be expressed in the IMCDs of young mouse kidneys (76). In the current study in mature rats, the KLF12 intensity on the arrays was, however, below background. Other KLF transcription factors are expressed in the IMCD including KLF5, KLF6, KLF9, and KLF10.
Rat GATA3 but not GATA2 can be amplified by RT-PCR from the collecting ducts of rat kidneys (89). Overexpression of rat GATA3 in primary culture of mouse OMCD cells can increase AQP2 activities, indicating that GATA3 could interact with GATA motifs in the AQP-2 promoter (89). Also, both GATA2 and GATA3 were detected at high signal intensity in the IMCD, but not in the non-IMCD portions of the inner medulla (Table 14B). GATA1 signal intensity was below background. Sp1 is also a member of the KLF transcription factor family. In the present study, Sp1 showed a moderate signal intensity in the IMCD.
TonEBP (Nfat5) is a Rel/Dorsal transcription factor that regulates cellular accumulation of organic osmolytes and HSP70. These mechanisms are critical to the process that protects cells from the hypertonic stress of the renal medulla. In the present study, TonEBP was indeed found to be expressed in the IMCD at a transcript level, although, the signal intensity was below the median on the arrays (Supplementary Table S1). Mice deficient in the TonEBP gene display severe atrophy of the renal medulla, presumably because the cells failed to adapt to the hyperosmolality (44). In response to long term hypertonicity in mpkCCD cells, TonEBP mediates expression of UT-A and possibly AQP2 independent of vasopressin level (26). However, AQP2 was not found to be strongly regulated by local osmolality in the renal inner medulla (82).
We thank Tony Cooper, Nalini Raghavachari for help with the Affymetrix array hybridization and data reporting, Guozhong Ma for setting up the online database, and Peter Munson and Xiuli Xu for advice regarding normalization of array data. This study was funded by the Intramural Budget of the National Heart, Lung, and Blood Institute (Project HL001285, M.A. Knepper, principal investigator).
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Blushing Cranberry Cider
Total Time
Prep 5 mins
Cook 30 mins
This is a lovely hot cider...served in a warm mug and garnished with orange slices. It is a delicious treat, after spending time out in the cold.
Ingredients Nutrition
- 1 blood orange, blood
- 1 cup unsweetened pineapple juice, pineapple, unsweetened
- 1 quart cranberry juice, red
- 2 cinnamon sticks, 3-inch pieces
- 1 piece fresh ginger, 2-inch piece, fresh, peeled and sliced lengthwise
- 3 whole allspice, balls (not ground)
- 2 cloves, whole
- 2 tablespoons dark brown sugar, dark brown (or to taste)
- 4 orange slices, thin (garnish)
- Remove two, 3-inch-long strips of peel from the orange and set aside until needed.
- Next juice and strain the orange.
- To a medium pot, add the orange, pineapple and cranberry juices, orange peel, cinnamon sticks, ginger, allspice and cloves.
- Make sure to sample and add the sugar if desired.
- Heat over medium-high heat until it starts to simmer, then reduce the heat to low and continue to simmer 30 - 45 minutes.
- Strain the cider, pour into warm mugs, add orange slices and serve immediately.
Most Helpful
Made this for 3 of us & doubled the recipe, since I wanted a bit left over to have COLD the next day, & as much as we enjoy a hot cider, it was every bit as great tasting chilled as it had been warm! Absolutely wonderful tasting & a great keeper of a recipe! Thanks for sharing it! [Made & reviewed in 1-2-3 Hits Tag]
What a delicious cider. I used a sugar free cranberry juice and omitted the blood orange since there weren't any available in the supermarket. The rest of the recipe was prepared as written. It makes a lot more than 2 servings so I'll be able to warm some up to enjoy later this week. Made for Photo Tag.
A clever and DELICIOUS drink! It takes a little preparation time, but it is worth it! I used pineapple nectar and cranberry nectar, which is thinner than 100% juice - but still great for this mixed drink. My only sub was NO orange slices - I had used all my oranges in my home-made Christmas cake and mincemeat! This was really lovely - Christmas in a mug, and it is on my menu for the Festive period. I plan to make it before I need to serve it, and then heat it up gently on my woodburning stove! AMAZING, it's the spices that make this so tangy and different to other hot cider drinks! Thanks BK! Made as a special Fairy Dust thank you for MsSally in Holiday Tag. FT:-)


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Peyton Godwin recorded the fastest time at the first "Flash Dash," a 5K race that began Saturday's Florida Catholic State Championship action at St. Joseph Academy.
Godwin, 48, led the race with a time of 18 minutes, 43 seconds.
"We did something new," said Judge John Alexander, who has been around for the championship at St. Joe in the past and was a big part of its current revival. "It turned out really well. We're making this a community event a week before Christmas."
Molly Burkhardt, formerly of St. Augustine, had a dominant performance for Bishop Kenny in its 3-1 (25-19, 25-22, 23-25, 25-19) win over Bartram Trail.
Sarah Ragland led the Bears (0-1) with 13 kills.
Bailie Burkins added 27 digs. Savannah Murray had 8 service points and 5 aces. Nicole Williams recorded 3 blocks.
Boys golf
Bartram Trail recorded its second win of the season (2-0) with a victory over Nease, 143-153.
Medalists for the Bears included Andrew Alligood (34), Jordan Batchelor (36), Harrison Presta (36) and Jackson Dunlap (37).
St. Johns County Fire Rescue will decrease lifeguard staff on county beaches starting Sept. 3, marking the end of the summer beach season.
There will still be mobile truck patrols through the various beach zones, until mid-September, though lifeguard towers will no longer be staffed.
A 30-year-old San Mateo man was hit and killed and a woman injured by a southbound passenger train at about 7 a.m. Thursday in Putnam County, according to the Florida Highway Patrol.
A release from FHP identified the man as Stephen Carl Smith. The woman, 32-year-old Amanda Harvey of East Palatka, was taken to Shands Gainesville with serious injuries.
Sgt. Hancel Woods of the Putnam County Sheriff's Office said the accident happened at the rail intersection with County Road 309 in Satsuma.
Creekside opened the volleyball season with a 25-9, 25-10, 25-17 victory over Fernandina Beach.
Allison Davanzo had nine kills and one block and Rachel Gupton had 22 assists and seven digs to lead the Knights.
Sophie deJong had seven aces, four kills and one block, and Mackenzie Lary had six digs and seven aces for Creekside.
Nease 3, Middleburg 2 (19-25, 25-17, 21-25, 25-14, 15-10): The Panthers started their 2013 season with a hard-fought win over Middleburg.
The City of St. Augustine has released an updated map of the shuttle service that it will provide during the Gentlemen of the Road concert tour stop Sept. 13-14 in St. Augustine.
Click here to see the updated map
The map shows six different shuttle routes with a total of nine pick-up/drop-off points.
Most of the routes will have just one destination other than Francis Field, which is the site of the event.
Local authorities conducted a patrol focused on stopping impaired drivers Saturday night, making five DUI arrests and writing 28 citations.
Eight officers from the St. Augustine Police Department, the St. Augustine Beach Police Department and the St. John's County Sheriff's Office helped, as part of the Labor Day traffic initiative, "Drive Sober or Get Pulled Over."
The six-hour patrol covered the areas near A1A/Beach Boulevard, Anastasia Boulevard and State Road 312, as well as other parts of the county.
It only took 12 minutes for police to thwart an attempted bank robbery Monday.
St. Augustine Police Department officers arrested James Jordan King, 35, as the suspected robber of the downtown Bank of America at 24 Cathedral Place, according to a SAPD release.
The officers responded to a bank alarm at 2:19 p.m. They quickly established a perimeter around the area and received a description of the man from bank employees.
The St. Johns County Sheriff's Office has released information about a convicted sexual predator who is now living in the county.
Registered sexual predator James Lee Lyons. will be temporarily residing at 7548 U.S. 1 South, the St. Johns County Sheriff's Office announced Monday.
The vehicle has been rendered safe. The SJSO Bomb Squad completed the search and roadways are now being opened.
Jacksonville Players Elite 18U, a travel softball team comprised of local players, won the 2013 Fast National tournament held at the Softball Olympic National Training Center in Clermont on Saturday.
The tournament took place over one week and featured softball teams from across the country. Jacksonville Players Elite went 9-0.
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On The World, Jane Hutcheon discusses the expectations on Benedict's successor with Bishop of Parramatta Anthony Fisher and ABC Religion's Scott Stephens.
Topics: religion-and-beliefs, religion-and-beliefs, catholic, australia
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People in the area worst affected by the Fukushima nuclear accident have a higher risk of cancer, the WHO says.
Topics: nuclear-accident, disasters-and-accidents, health, japan
Russia has unveiled the draw for what is set to be a fiercely-contested Rugby World Cup Sevens in Moscow.
Topics: rugby-7, rugby-union, sport, russian-federation
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Australia and Japan's war over whaling is heating up with Japanese whaling fleets and Australian anti-whaling ships clashing at sea. Japan has declared there will never be an end to whaling, but Australia continues to fight them in the courts.
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Both the Malaysian Government and Opposition are attempting to secure the youth vote as a general election must be held by June. Younger generations now make up a substantial part of the voting public, with social media playing a bigger role than ever before.
Space flight analyst Morris Jones says an American multi-millionaire's plan to send a couple on a mission to Mars is too ambitious and won't work.
Topics: astronomy-space, space-exploration, spacecraft, australia
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Asia contributed 40% of the world's total growth last year with China playing a big part in the economic boom. The World Bank says the economies of the Philippines and Thailand are contributing to the growth, and Burma is showing enormous potential. Kesha West spoke with Axel Van Trotsenburg, the World Bank's new Vice President for East Asia and the Pacific.
Topics: business-economics-and-finance, asia
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A group of filmmakers, musicians and surfers gathered on the North West coast of Australia to create 'The Reef', a unique celebration of surfing and music. The film is now being recreated in concert with the help of the Australian Chamber Orchestra.
Topics: opera-and-musical-theatre, australia
Indonesia's most influential Islamic organisation has complained to the national police chief over mistreatment of terrorism suspects.
Topics: terrorism, law-crime-and-justice, police, indonesia
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Retail profit results today highlighted an environment of winners and losers. The non-discretionary kings, supermarket chain Woolworths, recorded a lift in profits, but Harvey Norman was hit by the savage discounting in its electronics business. But both players signalled better times ahead.
Topics: business-economics-and-finance, australia
Posted| Updated
Step aside in the Senate and let us get on with scrapping the carbon tax. That is the warning from the Coalition to Labor if it wins government in September. The Shadow Climate Change minister says business is on side with axing the tax.
Topics: business-economics-and-finance, environment, australia
Posted| Updated
David Buik from Cantor Index joins Ticky from London.
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A fall of over 36 per cent to $82 million in net profit for the first half year, reflected a tough trading period for Harvey Norman with aggressive discounting and losses on property development. But it was an ebullient CEO Gerry Harvey who spoke with Ticky.
Topics: retail, business-economics-and-finance, australia
Posted| Updated
Tony Abbott outlines his higher education agenda, highlighting growth in online courses and reducing red tape for overseas students.
Topics: university-and-further-education, education, liberals, political-parties, government-and-politics, australia
Posted| Updated
Friday marks the final weekday broadsheet editions of The Sydney Morning Herald and The Age. Industry watchers warn that the move to a compact size won't necessarily arrest the long term decline of print media or win over sceptical advertisers.
Topics: business-economics-and-finance, print-media, australia
The Liberal Party has promised to create a 20,000 square kilometre national park and a 30,000 square kilometre marine park in the Kimberley, if it wins the State election.
Topics: national-parks, kununurra-6743
Shane Watson expects a spin-friendly wicket at Hyderabad for the second Test against India starting on Saturday.
Papua New Guinea's police force has revealed it has dismissed 87 officers for misconduct in the past two years, half of them for police brutality.
WA ALP Announcement
A Bangladesh tribunal has sentenced an Islamist party leader to death for abuses during the country's 1971 independence war.
Topics: rights, human, bangladesh, asia
Posted| Updated
A police officer charged with speeding while escorting singer Jimmy Barnes to a concert in Western Australia's south-west has been fined $700.
Topics: traffic-offences, law-crime-and-justice, courts-and-trials, police, margaret-river-6285, wa, australia
Posted| Updated
In this episode, the Mariners ready to burst Western Sydney's bubble in Saturday night's A-League blockbuster, Casey Stoner's promising start to his career on four wheels and Peter talks to retiring Australian cricketer Lisa Sthalekar.
Two different groups of election observers have delivered similar, damning assessments of last year's national elections in PNG.
Topics: federal-elections, corruption, papua-new-guinea, pacific
Two different groups of election observers have delivered similar, damning assessments of last year's national elections in Papua New Guinea.
Topics: corruption, federal-elections, papua-new-guinea, pacific
Ashleigh Barty reached her first ever WTA quarter-final with a straight-sets victory over Zarina Diyas in Malaysia.


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Professional cowboy Cam Billing, of Te Awamutu, remains in a
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the Otago Regional Rescue Helicopter to Dunedin Hospital and
later transferred to Burwood, in Christchurch.
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Harley Quinn's Greatest Moments from "Batman: The Animated Series"
TV, Comic Books
News and reports from the New York Comic Con rolled out even after the lights were turned off on Sunday; here are a few of them, as well as some tidbits we missed the first time around:
• Marvel announced an ongoing Age of Apocalypse series by David Lapham and Roberto De La Torre, spinning out of the current "Dark Angel Saga" storyline in Uncanny X-Force. [CBR]
• Designer Chip Kidd is writing a Batman book called Batman: Death by Design with art by Dave Taylor. It's due out next summer. [ComicsAlliance]
• USA Today spotlighted Captain Brooklyn, due next May from Jimmy Palmiotti, Frank Tieri and Amanda Conner. The three-issue miniseries will be published by Image Comics. [USA Today]
• Following the convention, Marvel has released pages from the Prep & Landing story that will appear in a few of their upcoming November comics. [CBR]
Saturday at the New York Comic Con brought news for the Avengers, Superman, Legendary Comics and … Disney's Prep & Landing? Here's a round-up of announcements from the show today.
• With a big, blockbuster Avengers movie scheduled for next May, Marvel announced a new ongoing series, Avengers Assemble, by writer Brian Michael Bendis and artist Mark Bagley. The book will launch next March and will feature most of the Avengers featured in the movie — Iron Man, Captain America, Black Widow, Hawkeye and the Hulk. The first arc will feature the villainous group the Zodiac.
• Speaking of that big, blockbuster Avengers movie, fans were treated to new footage from it featuring Bruce Banner and the Black Widow. Tom Hiddleston spoke to CBR about his work on the film.
• Marvel also announced that writer Rick Remender and artist Gabriel Hardman will take over Secret Avengers with issue #21.1, adding new members and pitting them against a new Masters of Evil.
• At the Cup O' Joe panel today, Marvel also announced a Disney/Marvel crossover — Prep & Landing: Mansion: Impossible. It features the elves from the Disney television special who prepare homes for the arrival of Santa Claus every Christmas eve — only this time they're trying to break into Avengers Mansion to get it ready for Santa. Written by director Kevin Deters and drawn by story artist Joe Mateo, the story will run in the back of the Marvel Adventures books as well as Avengers #19 in November.


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Mass Effect 3 Ruthless Enemies Trailer Analysis – IGN Rewind Theater
Update: Senior designer at BioWare Brenon Holmes have confirmed Brutes are Krogan/Turian hybrids. twitter.com Want to see some of the new enemies in Mass Effect 3? Delve a little deeper with IGN editors Destin Legarie and Sam Claiborn as they reveal details you may have missed in the latest trailer highlighting the enemies of Commander Shepard in this special IGN Rewind Theater. Release Date: March 6, 2012 Also on: PC, PS3, Xbox 360 Rated: M Genre: RPG Publisher: Electronic Arts Developer: BioWare Online Play: 4 Co-op Grab your omni blade and head to IGN for more ME3: xbox360.ign.com Subscribe to IGN's channel for reviews, guides, trailers and all things gaming: www.youtube.com
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'Snow White and the Huntsman' 2012 – Official Trailer in HD!
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« Exclusive Hit and Run TV Spot Rupert Sanders' Wife: You Will Not Direct Sequel To Kristen Stewart Movie! »


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First Avenue's 20 best concerts: The complete list
First Avenue: An archived history via flyers, riders & contracts
First Avenue's 20 best concerts: Readers' picks
First Avenue's 40 years of rock 'n' roll
Top 20 best Minnesota musicians: The complete list
First Avenue is undoubtedly among the premier rock clubs in the country -- possibly the world. Since 1970, the building at the intersection of First Avenue and Seventh Street North in downtown Minneapolis has hosted drinking, dancing, and live music as the Depot, Uncle Sam's, Sam's, and since 1981, First Avenue. The iconic silver stars naming past performers painted on the outside walls tell part of the story of the club, but the anecdotes of obscenely cheap tickets, balmy temperatures, and youthful days arriving long before the jaded regulars are what make this building an institution. Even riskier than summing up the best shows from the club's 42-year history would be to let these ear-shattering, heart-warming evenings go forgotten. Here are 20 of the finest nights First Avenue ever had. --Reed Fischer
20. Arcade Fire/ Wolf Parade/ Bell Orchestre, First Avenue, 9/29/05
Arcade Fire were well on their way to indie-rock success story when they rolled into this sold-out Mainroom show, but even the local skeptics became full-blown believers. The band played nearly all of their utterly brilliant debut LP, Funeral, as well as three tracks from their self-titled EP, but it was their stellar cover of David Bowie's "Five Years" -- the band had just performed it with him earlier that month at a Fashion Rocks event -- that truly made this show magical. By the time openers Wolf Parade came out to join the band for a euphoric version of "Wake Up" that closed out the show, the Canadian collective had alerted each and every one of us that a new day was indeed dawning in rock 'n' roll. --Erik Thompson
19. Meat Beat Manifesto, 7th St. Entry, 11/10/89
18. Burning Spear, First Avenue, 9/24/01
Reggae legend Burning Spear (Winston Rodney) has always been a powerful performer at First Avenue. But in the first weeks following 9/11, reggae's pillars of peace, love, and positivity seemed like Dorothy believing in the Wizard. If anyone else in the Mainroom was doubting the love of Jah Ras Tafari that night, Spear shook it out of us. He was in very strong voice, and his band was coming off of months on the road together: They could feel each other. Spear fixed his eyes at a point above and beyond the audience and swayed, his shoulders shook with a shiver and he started, "We are here to pick up the pieces, come along my brothers, come along my sisters." He seemed to be channeling more than singing that night. My friend took pictures and she swears there are "orbs" around Burning Spear in the photo. I don't believe in orbs, but I believe in reggae again.--Rachel Lee Joyce
17. Pulp, First Avenue, 5/26/96
It's stunning how much the nattily dressed Jarvis Cocker expressed with his legs during this performance. The Pulp frontman thrilled a lively, combative crowd while leading his band through a set that pulled heavily from the near-perfect Different Class album. While every Britpop fan at the time (including myself) was wasting far too much breath on Oasis, this was the band that exceeded what little stateside hype they'd earned. One woman in the crowd was either trying to flirt with Cocker, or just make a scene. Either way, she repeatedly lambasted him for the commentary found in "Common People." And, proving his deftness once again, he sidestepped it all and suggested they discuss the matter afterward. And a not-so-humble brag to those who are shelling out to see Pulp in reunion mode right now, note that this show was $5! --Reed Fischer
16. Erykah Badu, First Avenue, 4/30/97
Erykah Badu's debut release Baduizm had not yet received much play on mainstream radio ahead of her first full U.S. tour, but the club was still packed that night. For a change, the sweet-scented swirls of smoke weren't coming from the crowd -- Ms. Badu had filled the stage with incense, tapestries, and floor lamps with colored bulbs, which were the requisite trappings of a neo-soul session back then. She captivated us with the drawl-tinged octaves of her voice so much that even her between-song lectures in Egyptian numerology and African spirituality were met with air toasts to the stage. The jazzy/blues from the record translated perfectly with a full band, a rarity at R&B shows of the era. She mostly stuck to tracks from Baduizm, but when she introduced the unreleased "Tyrone," we seemed especially connected as we we sang the chorus and, when instructed by the true diva, pointed at our "third eyes." --Rachel Lee Joyce
15. PJ Harvey, First Avenue, 10/01/04
While PJ Harvey's 2004 album Uh Huh Her didn't quite match Mercury Prize-winning Stories From the City..., her First Avenue tour stop in support of that record was memerizing. In a full-length red velvet gown and matching gloves, Polly Jean tore through her stellar catalog while confidently delivering performances of "Dress," "Good Fortune," "50 Ft. Queenie," and "Meet Ze Monsta." Harvey was also still wailing away on her guitar, this being the era before her eventual foray into the stark piano and harp-laden arrangements of her current work. --Erik Thompson
Photo by Steve Cohen
14. Lucinda Williams, First Avenue, 7/17/01
13. Marilyn Manson, First Avenue, 10/09/96
At the time of this show, I was 15 years old, and had recently parted with a narrow and pinched childhood by becoming a goth. This rebellion was deflated a little by the fact that it was endorsed by my mother, who believed so deeply in broad-mindedness that she sprang for my ticket, fairly ruining the main thrill of it. The show was magnificent, but the venue captivated me the most. Its soul was stranger and more beguiling to me even than Manson's, and though my affair with Manson's music, torrid as it was, proved fleeting, my affair with First Avenue has seemed star-crossed from the first. --David Hansen
Photo by Steve Cohen
12. The Flops/ Dan Wilson & Friends/ Semisonic/ 3/4 Trip Shakespeare 12/12-13/03
11. Dinosaur Jr./ My Bloody Valentine/ Babes in Toyland, First Avenue, 2/12/92
My Bloody Valentine was touring on Loveless, and splitting the bill with Dinosaur Jr. In this case, they were the middle act. I don't know anyone who stayed, but it was a show that bordered on the mystical. The club literally ran out of earplugs. I have had mild tinnitus ever since, and I don't care -- it was glorious. I was awash in sound. --Chris Strouth
10. Basement Jaxx, First Avenue, 10/08/01
Basement Jaxx's
9. Pavement/ Wilco/ The Dirty Three, First Avenue, 05/26/95
There are some headlining sets that are so good that no matter who the openers are, you can't remember much about their performances. That is clearly the case with Pavement's riveting Mainroom show in 1995. Wilco were there in support of their just-released debut, A.M., along with frequent Nick Cave collaborators the Dirty Three, while Pavement were touring behind their brilliant new record Wowee Zowee. As with all of Pavement's live performances, this was a splendidly shambolic show that drew generously from all three of their stellar LPs. The band were still getting along relatively well at this point, which gave their underdog anthems an irresistible slacker spirit that the college-aged crowd clearly identified with. "Rattled By The Rush," indeed. -Erik Thompson
8. The Fugees/ The Roots/ Goodie Mob, First Avenue, 11/96
Three of the era's biggest acts hit the road on a small club tour to rave reviews, but First Avenue was the high point as each crew wanted to out-rock each other in the house that Prince built. The Fugees headlined, and it just as "Killing Me Softly" was about to repeat on pop radio. This was Lauryn Hill at her best. Her voice boomed off the walls as Wyclef Jean cemented himself as a true performer and, oh yeah, Pras was in there somewhere. The Roots were on-point creatively, and fresh with their live band and impromptu jam session. Black Thought did covers of Wu-Tang, and impersonated each member perfectly. You swore he was magically channeling Ghostface. But no doubt this new group from Atlanta had everyone talking, Goodie Mob stole the show with a high-energy charismatic performance. Cee-Lo rapping his full verse on "Goodie Bag" brought the house down. People who were in attendance knew this was a special night and knew something like this would never happen again as Khujo starting throwing buckets of fried chicken into the audience as an encore, we were all celebrating this golden show during this golden era of hip-hop. -Lars Larson
7. Rhymesayers Entertainment 10th Anniversary Show, First Avenue, 7/22/05
Like an early Motown Records revue, not one group was the headliner. Instead, it was the whole label, or in this case, a whole city and movement. Rhymesayers celebrating their 10th anniversary was a "we arrived"-type moment and the crowd knew it as well. From the early days in the Entry to cats, vans, bags across the country, it was time to cut the cake and drop the balloons. The venue was packed to the doors and stifling hot. But the fire came from the stage as the show was fast paced with no downtime or lull and every act from Atmosphere to Musab brought its A game with short, powerful sets. Los Nativos charged the crowd in full headdress and war paint attire and new label mates P.O.S and Mac Lethal proved they belonged. Everyone got stage time, even a real MF Doom managed to show up, and you know it's a special night when that happens. -Lars Larson
6. The Jayhawks, First Avenue, 7/7-8-9/95
5. U2, First Avenue, 2/21/82
In a smaller venue, the young and spiritually charged rising rock stars U2 were mixed with a devoted audience that, even then, knew every word to every song. Literally, as Bono struggled with the words to a "Southern Man" encore, he brought up a member of the audience to help him with the performance. This was Bono before the bug shades, this was Edge debating the relationship between faith and the rock 'n' roll lifestyle, and this was way before multi-million dollar Pop Mart stage setups. This was an simple Irish band, up close and personal with its fan base, they would never be this intimate and vulnerable again. The electricity of this night will never be matched again as U2 moved on to bigger venues over the years. -Lars Larson
4. Radiohead, First Avenue, 4/03/96
Many Radiohead devotees have already spoken out about the legendary show the band put on the year prior to this gig (6/04/95, although my ticket stub says 6/11/95). In both cases, a brash, young folk singer named David Gray was the opening act, and both shows were examples of the band right before meeting people became so easy. Having attended both, the 1996 concert had more of a feel for where the band would end up on OK Computer. For Bends purists, this would be a nightmare. But those of us looking for the band to distance itself from the Buzz Bin, hearing wild renditions of songs like "Ripcord" that would soon go in the vault forever was only the tip of those Kid A icebergs. Plus, they did an in-store signing at Let It Be Records after the concert. -Reed Fischer
3. Public Enemy, First Avenue, 12/12/88
In a time before Chuck D became the hip-hop guy who every NPR listener could quote, Public Enemy was the band that was scary. Hip-hop went from the cartoony plumage of Grandmaster Flash to the hardcore gangster attitude of Ice-T and PE. Walking into their first show felt dangerous -- at least from my perspective as gothy teenager from Fridley. It felt like a moment where everything changed. Punk rock wasn't dangerous anymore, but this was. With Griff and the hard-stepping of the Nation of Islam soldiers, Chuck D and Flavor Flav stepping so hard that it was bouncing Terminator X's turntables. It was a bit like losing your virginity, only without sex and a lot more references to Louis Farrakhan. -Chris Strouth
2. Prince, First Avenue, 7/07/07
Needless to say, there are countless Prince shows at First Ave that could fit in this slot. Take your pick from the run of mid-'80s blowouts, the filming of Purple Rain, the exploration of Sign O' the Times, and so forth. This was the most surprising show of them all. On the morning of 7/7/07, my alarm went off at 5 a.m. I rolled out of bed and got hastily dressed. It was Prince day. People had been sleeping on the sidewalks surrounding First Avenue since the night before, and I was worried I wouldn't get there in time. After an eight-hour wait I had both a wristband and a ticket.
Another eight-hour wait later that night proved that it was worth everything to see Prince onstage in his second home again. It came along with a performance in Macy's and at Target Center. Sheila E and Wendy showed up, among other special guests, and it took the Minneapolis police to shut down the show around 4 a.m. -Stacy Schwartz
1. Joe Cocker, The Depot, 4/03/70
Nothing feels as sweet as the first time. According to the April 4, 1970 edition of the Minneapolis Tribune, "not since the truck drivers' strike of 1934 is it likely that there has been such excitement, such chaos, such congestion, such noise just off Hennepin Av. as there was Friday night." The account, which later calls the space Fillmore Upper Midwest, says that carpeting and other interior decorations weren't yet installed, but the old bus station was packed out with sun-tanned men and women wearing expensive hippie garb by the time Cocker hit the stage just after 8 p.m. The $10 tables were full, the $4 standing room was reserved for those willing fight for it, and the initial stocking of booze was used up by showtime. A long-haired Cocker worked hard on stage filled with 40 people "singing like a black man" and dancing "like a spastic" and the two sets had the feel of a circus sideshow. From day one, this club had its magnetic persona established. -Reed Fischer
What was your favorite show at First Avenue? Write us, and send photos to [email protected] and we'll include you in a list of readers' picks on the blog next week.
First Avenue: An archived history via flyers, riders & contracts
First Avenue's 20 best concerts: Readers' picks
First Avenue's 40 years of rock 'n' roll
Top 20 best Minnesota musicians: The complete list
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my Sitemap, which is generated by XML Sitemap & Google News Feeds Plugin, is not working. Problem is that I have about 25.000 Posts stored in my WordPress Database - the sitemap just keeps loading and loading.
AFAIK, Quick Cache is supposed to help with WP installations with this size of content, it even says inside the Sitemap Plugin that Quick Cache is recommenend.
My Question now is, how can I enable the Sitemap Cache, since it seems not to be working?
Or if there is no "activate" switch for this, how can I find out why its not active already? Any typical problems that match to this case + solutions?
I already "splitted by month" inside the xml plugin - didnt help.
Thanks for the help in advance


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As for old age, embrace and love it. It abounds with pleasure, if you know how to use it. The gradually (I do not say rapidly) declining years are amongst the sweetest in a man's life; and, I maintain, that even where they have reached the extreme limit, they have their pleasure still.
Seneca (4 BC - 65 AD)
Contributed by: Zaady
The happiness of life is made up of minute fractions - the little, soon forgotten charities of a kiss or smile, a kind look, a heartfelt compliment, and the countless infinitesimals of pleasurable and genial feeling.
Samuel Taylor Coleridge (1772 - 1834)
In Xanadu did Kubla Khan A stately pleasure-dome decree: Where Alph, the sacred river, ran Through caverns measureless to man Down to a sunless sea.
Source: Kubla Khan
What is written without effort is in general read without pleasure.
Samuel Johnson (1709 - 1784)
The great source of pleasure is variety.
When a man says he had pleasure with a woman he does not mean conversation.
"I fly from pleasure," said the prince, "because pleasure has ceased to please; I am lonely because I am miserable, and am unwilling to cloud with my presence the happiness of others."
Source: Abyssinia. Rasselas. Chap. iii.
The natural flights of the human mind are not from pleasure to pleasure, but from hope to hope.
The expense is damnable, the position is ridiculous, and the pleasure fleeting.
His death eclipsed the gayety of nations, and impoverished the public stock of harmless pleasure.
Source: Life of Edmund Smith (alluding to the death of Garrick).
Copyright © 2016 Gaiam, Inc.


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Protecting Your Plants from Frost and Freeze Damage
1. Temperature dropping to 28 degrees is considered a light frost, or light freeze, which usually
affects tender plants such as annuals. It may also damage early blooms and leaves on deciduous shrubs and trees.
2. Temperatures of 25-28 degrees are considered a hard freeze and will most likely affect foliage and blossoms.
3. April 15th is usually our last freeze date. It is best to wait until after this date to plant tender
annuals and vegetable crops.
4. If you have tender plants planted, perennials with tender growth emerging, or trees and shrubs with tender blossoms, such as camellia, you can do one of several things:
Water thoroughly before nightfall.
Cover plants with old blankets, sheets, burlap or cardboard boxes. Drape loosely so as not
to crush plants. Remove covers as soon as the sun comes up.
Water plants in pots. Wet soil holds more heat than dry soil.
Mulch low tender plants with straw, pines needles, bark or loose leaves.
5. If you are unable to move container plants, where roots are more susceptible to freezing, indoors or under cover, you may wrap the pot in burlap or bubble wrap, or bury the pot in soil in addition to protecting the foliage.
6. If you do get some damage from frost, do not prune your plant until the weather warms up and new growth comes out. Most times the plant will regenerate new leaves, but flowers will probably be lost, as will the fruit that the flower would have produced.
7. If you are worried about losing blooms, such as forsythia and quince, cut some branches and bring them inside for your enjoyment. Otherwise, delay pruning until danger of frost is past.
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Yes, it is very scary having a gun pointed at you.
Since you weren't the one driving your options were limited to praying or shooting, unless you decided to bail out of the car to seek cover and or engage in a gunfight. I won't tell you what I would've done, but I will give you some options, beyond those I've already typed.
Being that you're in a 2,000 pound bullet, the car, that also provides a way to flee the scene to safety I wouldn't be suprised if the gun holder becomes a speed bump as I am getting my butt to safety if he was standing in front of the vehicle. I'm not a fan of engaging in gunfights when I have a better felon stopper at my disposal. I'd rather undergo an investigation for running somebody over in defense of my life than a shooting/homicide investigation.
Shooting from a moving vehicle is difficult, and often results in misses that you are responsible for. Heck, you are responsible for every bullet you fire, misses and solid hits. There is no way to tell, from your post, if his amigos were acting in concert with him. Even if they were, you can bet that those that survive will say they weren't.
Shooting through window glass, especially the front window, tends to deflect the bullets path. Remember, the bullet must pass through a curved surface and then continue to the target. So, my question to you is, do you know if your bullet will deflect upward or downward from where you would shoot from? Also, after passing through that barrier, will hit hold together well enough to stop the threat and penetrate deeply enough?
Just some food for thought. I will also state that pointing a gun at me places me in fear of my life, and meets the legal requirement for using lethal force in the described scenario.


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Heading into regionals this weekend, Manhattan High girls' tennis coach Joyce Allen likes the way her team has been playing lately.
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text: I'm having a pretty nasty problem with wordpress. Looks like that after a certain number the remaining characters are cut off in sidebar. So whatever I put into sidebar.php, after a certain number of characters it's cut off. Is there a workaround for this or anything else that could be done about this?