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Allosteric inhibition enhances the efficacy of ABL kinase inhibitors to target unmutated BCR-ABL and BCR-ABL-T315I
Infection, immunity and cancer vaccines
Chronic myelogenous leukemia (CML) and Philadelphia chromosome-positive (Ph+) acute lymphatic leukemia (Ph + ALL) are caused by the t(9;22), which fuses
The efficacy of this approach on the leukemogenic potential of BCR/ABL was studied in Ba/F3 cells, primary murine bone marrow cells, and untransformed Rat-1 fibroblasts expressing BCR/ABL or BCR/ABL-T315I as well as in patient-derived long-term cultures (PDLTC) from Ph + ALL-patients.
Here, we show that GNF-2 increased the effects of AKIs on unmutated BCR/ABL. Interestingly, the combination of Dasatinib and GNF-2 overcame resistance of BCR/ABL-T315I in all models used in a synergistic manner.
Our observations establish a new approach for the molecular targeting of BCR/ABL and its resistant mutants using a combination of AKIs and allosteric inhibitors.
Chronic myeloid leukemia (CML) and 30% of adult acute lymphatic leukemia (ALL) are characterized by the Philadelphia chromosome (Ph+), which is the cytogenetic correlate of the (9;22) chromosomal translocation. The t(9;22) leads to the fusion of the
Selective targeting of BCR/ABL by ABL-kinase inhibitors (AKI) such as Imatinib, Nilotinib or Dasatinib, all competitive ATP-analogues, leads to durable cytogenetic and molecular remissions in the majority of CML patients in the early chronic phase of the disease. However, unsatisfactory responses in advanced disease stages, resistance and long-term tolerability of BCR/ABL inhibitors represent major clinical problems. In fact, advanced CML and Ph + ALL respond only transiently to AKIs
The activation status of wild-type c-ABL is finely regulated by several regulation signals. Myristoylation of the N-terminus of c-ABL is involved in the regulation of the ABL kinase activity. The N-terminus of ABL is myristoylated, and the myristate residue binds to a hydrophobic pocket in the kinase domain - the myristoyl-binding pocket (MBP) – in a process called "capping". The "capping" leads to conformational changes that allow the intramolecularly docking of the "SRC homology 2 domain" to the kinase domain. Hence, c-ABL adopts an auto-inhibited conformation. The absence of an N-terminal myristoylated domain activates c-ABL consistent with its auto-regulatory role. In the context of the t(9;22), the N-terminal auto-inhibitory "Cap" region is substituted by the BCR portion of the fusion protein. The absence of the "Cap" region allows the BCR/ABL to "escape" auto-inhibition contributing to the constitutive activation of its kinase activity
We have recently shown that the allosteric inhibition increases the sensitivity of BCR/ABL-T315I towards the inhibition of oligomerization most likely by interfering with the overall confirmation of the kinase
The cDNAs encoding BCR/ABL and BCR/ABL-T315I have been previously described (3). All retroviral expression vectors used in this study were based on the bi-cistronic PINCO vector.
Cell lines and patient-derived long-term cultures
The Ba/F3 and Rat-1 cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany) and were maintained as previously described (3). Ph + ALL patient derived long term cultures (PDLTCs) expressing BCR-ABL-T315I (KÖ) were obtained from a patient enrolled in the German Multi-Center Study Group for acute lymphatic leukemia of the adult (GMALL 07/2003) upon informed and written consent
Isolation of Sca1+/lin- hematopoietic stem and progenitor cells (HSPCs)
Sca1+/lin- HSPCs were isolated from 8- to 12-week-old female C57BL/6 N mice (Janvier, St. Berthevin, France) after euthanization by CO2 asphyxiation. Bone marrow (BM) was harvested from the femur and tibia by flushing the bones with a syringe and a 26-gauge needle. Sca1+ cells were purified by immunomagnetic beads using MACS cell separation columns according to the manufacturer's instructions (Miltenyi, Bergisch-Gladbach, Germany). Prior to subsequent use, the purified cells were pre-stimulated for 2 days in DMEM supplemented with 10% FCS (Hyclone/Perbio Science, Bonn Germany), 1% L-Glutamine, 1% Penicillin/Streptomycin, mIL-3 (20 ng/mL), mIL-6 (20 ng/mL) and mSCF (100 ng/mL) (Cell Concepts, Umkirch, Germany).
Transfection and retroviral infection
Ecotropic retroviral supernatants were obtained after transfection of Phoenix packaging cells as described earlier
Transformation assays
Soft agar and focus formation assays were performed using untransformed Rat-1 fibroblasts retro virally transduced with PINCO vectors harboring unmutated BCR/ABL or BCR/ABL-T315I. Six-well plates were filled with DMEM supplemented with 10% FCS and 0.5% bacto-agar (DIFCO Laboratories, Detroit, MI, USA) (2 ml per well). Then, 5x103 transduced Rat-1 cells were suspended in "top-agar" (DMEM supplemented with 10% FCS and 0.25% bacto-agar) (1 ml per well) and stacked in the wells. Colonies were counted after 15 days of incubation at 37°C and 5% CO2. For focus formation assays, 4x104 transduced Rat-1 cells were plated per well of a 24-well plate. Foci were stained after 15 days using 1% crystal violet (Sigma).
Colony assays on HSPCs
At day 5 post-infection, Sca1+ cells were plated at 5x103 cells/mL in methyl-cellulose either with mIL-3 (20 ng/mL), mIL-6 (20 ng/mL) and mSCF (100 ng/mL) or without cytokines (Stem Cells Inc., Cambridge, UK). The number of colony forming units (CFUs) was determined 10 days after plating and normalized according to the transduction efficiency.
Western blotting
Western blot analysis was performed according to widely accepted protocols. The following antibodies were used: anti-ABL (α-ABL) (St. Cruz Biotechnology, Santa Cruz, CA, USA), anti-phosphorylated ABL specific for the phosphorylated tyrosine residue 245 (α-p-ABL-Y245) (Cell Signaling, Boston, MA, USA), anti-BCR (α-BCR) (St. Cruz Biotechnology), anti-phosphorylated BCR specific for the phosphorylated tyrosine residue 177 (α-p-BCR-Y177), anti-Crkl, and anti-phosphorylated Crkl (Cell Signaling).
Statistical analysis
Differences in response rates towards different concentrations of a single inhibitor or inhibitors in combination were analyzed by Student′s t-tests. Statistical analyses were performed using the GraphPad Prism (GraphPad Software, San Diego, CA) software package. Evaluation of the character of the combined effects was performed according to the three dimensional model of Prichard and Shipman using MacSynergy software
The allosteric inhibitor GNF-2 improves the response of unmutated BCR/ABL to AKIs
Unmutated BCR/ABL can be efficiently inhibited not only by AKIs but also by allosteric inhibitors such as GNF-2 or GNF-5
Figure 1
Effects of the allosteric inhibitor (GNF-2) and Abl kinase inhibitor (Dasatinib) on Ba/F3 cells expressing unmutated BCR/ABL
Effects of the allosteric inhibitor (GNF-2) and Abl kinase inhibitor (Dasatinib) on Ba/F3 cells expressing unmutated BCR/ABL. (A) XTT assay using Ba/F3 cells expressing empty vector upon exposure to 0.3, 0.6, 1.25, 2.5 and 5 μM GNF-2 and 0.3, 0.6, 1.25, 2.5 and 5 μM Dasatinib. (B) XTT assay using Ba/F3 cells expressing BCR/ABL upon exposure to 0.1, 0.2 and 0.4 μM GNF-2 and 5, 10, 25, 50 and 100 nM Dasatinib. Proliferation status was determined by the metabolic activity of cells given by the reduction rate of XTT to formazan. The means +/- SD of triplicates from one representative experiment out of three performed are given.
Taken together, these data suggest that the inhibition effect of Dasatinib is enhanced by GNF-2 in cells expressing unmutated BCR/ABL.
The combination of GNF-2 with dasatinib efficiently abolishes the BCR/ABL-T315I-mediated factor-independent growth of Ba/F3 cells
The major clinical challenge in Ph + leukemia is the drug resistance due to the "gatekeeper" mutation T315I. T315I confers a nearly global resistance to all molecular therapy approaches that target BCR/ABL. Neither GNF-2 nor AKIs have any effect on cells transformed by BCR/ABL-T315I. To analyze whether the combination of allosteric inhibition with AKIs is able to inhibit BCR/ABL-T315I, we exposed Ba/F3 cells expressing BCR/ABL-T315I to increasing concentrations of Dasatinib (0.3, 0.6, 1.25, 2.5 and 5 μM) and GNF-2 (0.3, 0.6, 1.25, 2.5 and 5 μM). Cytotoxicity and proliferation were assessed by the XTT assay. Here, we show that only the combination of GNF-2 and Dasatinib inhibited BCR/ABL-T315I-dependent cell growth with a very high synergy index of 186 (calculated combination index)(Figure
Figure 2
Effects of the allosteric inhibitor (GNF-2) and Abl kinase inhibitor (Dasatinib) on Ba/F3 cells expressing "gatekeeper" mutation T315I
Effects of the allosteric inhibitor (GNF-2) and Abl kinase inhibitor (Dasatinib) on Ba/F3 cells expressing "gatekeeper" mutation T315I. (A) XTT assay using Ba/F3 cells expressing BCR/ABL-T315I upon exposure to 0.3, 0.6, 1.25, 2.5 and 5 μM GNF-2 and 0.3, 0.6, 1.25, 2.5 and 5 μM Dasatinib. Proliferation status was determined by the metabolic activity of cells given by the reduction rate of XTT to formazan. The means +/- SD of triplicates from one representative experiment out of three performed are given. (B) Western blot analysis of Ba/F3 cells expressing the indicated transgenes using antibodies directed against the indicated proteins. (C) Calculation of the combined effect by the MacSynergy program.
Taken together, these data suggest that the allosteric inhibition sensitizes BCR/ABL cells harboring the "gatekeeper" mutation T315I towards the ATP analogue Dasatinib.
The combination of GNF-2 and dasatinib inhibited the growth of Ph + lymphatic PDLTCs expressing BCR/ABL-T315I
Ph + ALL expressing BCR/ABL-T315I is not fully represented in cell lines. Therefore, we tested the response of PDLTCs from Ph + ALL patients expressing BCR/ABL-T315I to GNF-2 and Dasatinib. The PDLTCs were directly derived from BM cells of Ph + ALL patients cultured in a specific culture medium
Figure 3
Effects of GNF-2 and Dasatinib on PDLTCs of Ph + ALL patients expressing BCR/AB-T315I
Effects of GNF-2 and Dasatinib on PDLTCs of Ph + ALL patients expressing BCR/AB-T315I. Proliferation/citotoxicity was determined by the metabolic activity of cells given by the reduction rate of XTT to formazan. The means +/- SD of triplicates from one representative experiment out of three performed are shown. (A) XTT assay using Ph- PDLTCs (HP) upon exposure to 1.25, 2.5 and 5 μM GNF-2 and 125, 25 and 500 nM Dasatinib. (B) XTT assay using Ph + PDLTCs (KÖ) expressing BCR/ABL-T315I upon exposure to 1.25, 2.5 and 5 μM GNF-2 and 125, 250 and 500 nM Dasatinib.
In summary, these data show that the combination of allosteric inhibition and Dasatinib overcomes the resistance in primary PDLTCs from Ph + ALL patients harboring the BCR/ABL-T315I mutation.
The combination of allosteric inhibition and dasatinib is able to abolish the transformation potential of BCR/ABL-T315I
We have shown recently that GNF-2 inhibits the transformation potential of unmutated BCR/ABL but not of BCR/ABL-T315I in untransformed fibroblasts
Figure 4
Effects of GNF-2 and Dasatinib on Rat-1 cells expressing BCR/ABL-T315I
Effects of GNF-2 and Dasatinib on Rat-1 cells expressing BCR/ABL-T315I. (A) Transformation assays measuring colony formation in soft agar. Rat-1 cells were retrovirally transduced with the BCR/ABL-T315I construct and seeded at 104 cells/well on soft agar in six-well plates in the presence of 2.5 or 5 μM GNF-2 and 300 nM Dasatinib. After 15 days, the colonies were counted. The means +/- SD of triplicates of two representative experiments are presented. (B) Focus formation assays. A total of 4x104 infected Rat-1 cells were plated per well in 24-well plates, grown for 72 h to confluence and incubated for 12 additional days in the presence of 2.5 or 5 μM GNF-2 and 300 nM Dasatinib. The plates were then washed, dried and stained with crystal violet. One representative of each of the two experiments performed in triplicate is given (34X magnification).
These data indicate that the combination of allosteric inhibitors with AKIs inhibits the transformation potential of BCR/ABL-T315I.
GNF-2 cooperates with dasatinib to inhibit colony formation of hematopoietic stem and progenitors cells (HSPCs) harboring BCR/ABL-T315I in semi-solid medium
To further confirm the synergistic effect of the combination of Dasatinib and GNF-2, we extended our investigation to a model of primary murine hematopoietic stem and progenitor cells expressing BCR/ABL. We studied the effects of the drug combination on the colony formation by BCR/ABL cells in semi-solid medium in the presence or absence of cytokines (mIL-3, mSCF and mIL-6). We transduced Sca1+ HSPCs with BCR/ABL-T315I, and plated the cells in methyl-cellulose with increasing concentrations of GNF-2 and Dasatinib. As shown in Figure
Figure 5
Effects of GNF-2 and Dasatinib on HSPCs expressing BCR/ABL-T315I
Effects of GNF-2 and Dasatinib on HSPCs expressing BCR/ABL-T315I. Sca1+ BM cells were retrovirally infected with BCR/ABL-T315. Infected cells were then plated in semi-solid medium in the presence of 2.5 or 5 μM GNF-2 and 0.3 or 0.6 μM Dasatinib (A) without cytokines and (B) with cytokines (mIL-3, mIL-6 and mSCF) to determine the effect of the drugs on HSPC colony formation in semi-solid medium. Colonies were counted after 10 days. The means +/- SD of triplicates from one representative experiment out of two performed are given.
These data demonstrate that mHPSCs expressing the "gatekeeper" mutation T315I can be targeted efficiently by the combination of GNF-2 and Dasatinib.
The major therapeutic challenge in Ph + leukemia is to efficiently treat patients with BCR/ABL harboring the T315I mutation. The T315I mutation is the most resistant to inhibition because of a combination of several factors, including steric hindrance of drug binding, loss of a key hydrogen-bonding interaction with the T315 side-chain hydroxyl group exploited by Imatinib, Nilotinib and Dasatinib and potentially through increasing aberrant intrinsic kinase activity accompanied by aberrant substrate phosphorylation
GNF-2 and its analogues are non-ATP competitive ABL kinase inhibitors, which bind to the MBP in the kinase domain. It seems that the binding of GNF-2 to the MBP stabilizes the protein in an inhibited conformation resulting in a structural reorganization of ABL that disrupts the catalytic machinery located in the ATP-binding region
An additive but not synergistic effect was shown for the combination of Nilotinib with GNF-2 or GNF-5 on BCR/ABL-T315I-related resistance. The stronger effects may be attributed to the fact that Dasatinib, originally developed as a SRC-kinase inhibitor, not only inhibits the BCR/ABL kinase but also targets a broader range of kinases compared to Nilotinib, the spectrum of which is mainly limited to ABL, c-KIT and PDGFR
Our data further establish allosteric inhibition as alternative or additional molecular therapy approach for the treatment of Ph+ leukemia. In fact, it not only overcomes the resistance mediated by the "gatekeeper" mutation T315I but also increases the response of unmutated BCR/ABL to AKI. In the clinical setting, this feature could contribute to a more efficient use of AKI at a lower dosage in "normally" responsive patients and the possibility to further increase dosage in patients early in the progression of disease, in the absence of BCR/ABL mutations, for whom dosage escalation is still a therapeutic option.
The results presented here contribute to the further development of allosteric inhibition for the molecular targeting of both unmutated BCR/ABL and BCR/ABL harboring the multi-resistance mutation T315I.
Resistance and long-term tolerability of BCR/ABL inhibitors represent the major therapeutic challenge in Philadelphia Chromosome-positive (Ph+) leukemia. Advanced Ph + leukemia respond only transiently to ABL kinase inhibitors (AKI). Resistance is mostly caused by the acquisition of point mutations in BCR/ABL. The "gatekeeper" mutation T315I confers resistance against all available molecular therapy approaches. Conformational changes by allosteric inhibition increases the response of both unmutated BCR/ABL and BCR/ABL-T315I towards inhibition of oligomerization. Therefore we investigated whether the conformational changes induced by the allosteric inhibition also enhances the response towards the AKI Dasatinib in clinically relevant models of Ph + leukemia. Allosteric inhibition not only increased the response of unmutated BCR/ABL to Dasatinib but also contributed to overcome resistance of BCR/ABL-T315I in a synergistic manner in all models used.
Therefore allosteric inhibition may contribute to the optimization of the therapy of patients with both unmutated BCR/ABL or harboring resistance mutations such as the T315I.
Competing interests
The authors declare that they have no competing interests.
Author's information
Afsar Ali Mian, Anna Metodieva both authors have to be considered first authors.
Authors' contribution
AAM carried out the studies on transduced Ba/F3 cells and participated in drafting the manuscript. AM performed the transformation assays on Rat-1 fibroblasts and on the murine HSCs. SB generated the PDLTCs harboring the BCR/ABL-T315I and carried out the experiments on these cells. MK and NR participated in the elaboration of the data. JM and YN participated in the design of the study and performed the statistical analysis. OGO participated in the design and coordination of the study. MR conceived of the study and wrote the paper. All authors read and approved the final manuscript.
This study was supported by a grant from the German Research Foundation (DFG) to MR, JM and YN (DFG-RU 728/3-2).
Induction of chronic myelogenous leukemia in mice by the P210bcr/abl gene of the Philadelphia chromosome
A progenitor cell origin of myeloid malignancies
Targeting of the N-terminal coiled coil oligomerization interface by a helix-2 peptide inhibits unmutated and imatinib-resistant BCR/ABL
Oligomerization inhibition, combined with allosteric inhibition, abrogates the transformation potential of T315I-positive BCR/ABL
Mechanisms of resistance to STI571 in Philadelphia chromosome-associated leukemias
Flying under the radar: the new wave of BCR-ABL inhibitors
Regulation of the c-Abl and Bcr-Abl tyrosine kinases
Long-term culture of primary human lymphoblastic leukemia cells in the absence of serum or hematopoietic growth factors
A three-dimensional model to analyze drug-drug interactions
Allosteric inhibitors of Bcr-abl-dependent cell proliferation
Targeting Bcr-Abl by combining allosteric with ATP-binding-site inhibitors
The gatekeeper mutation T315I confers resistance against small molecules by increasing or restoring the ABL-kinase activity accompanied by aberrant transphosphorylation of endogenous BCR, even in loss-of-function mutants of BCR/ABL
Activation of tyrosine kinases by mutation of the gatekeeper threonine
Allosteric interactions between the myristate- and ATP-site of the Abl kinase
Target spectrum of the BCR-ABL inhibitors imatinib, nilotinib and dasatinib
Myristoylation and membrane binding regulate c-Src stability and kinase activity
Pre-publication history
The pre-publication history for this paper can be accessed here:


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Aang's been busy with meeting after meeting, paperwork on top of paperwork. He's sleepy, and Katara is going to make sure he rests...
Not every 15-year old has to contend with this load of work.
It's just a bit different when you're the Avatar and, although it had been three years since its end, you have a world still reeling from a war of one hundred years that ripped at every piece of its delicate balance.
So was the case for Aang.
Sitting in his icy South Pole home, the Avatar sifted through several scrolls; appeals from people all throughout the Earth Kingdom and Fire Nation, arguments about continued Fire Nation colonization in parts of the country. There was all the talk about adjusted trade routes and the formation of this so-called "fifth nation" which would comprise all the colonies.
Aang had to answer these people in debates at the peace meetings - which he just had two weeks of - and then afterward in these written appeals. He also had to figure out the proper plan for the world to balance itself.
So goes the Avatar's duties.
The young airbender had been up for three days - almost straight. He could sneak in an hour of rest here and there, but other than that, it had just been straight paperwork. He felt that he had to it for all the world's leaders and all the people he pledged to serve in his position.
However, one couldn't help but wonder when he could put his own basic needs into view. Sure, he had his duty to the world...but that didn't mean he couldn't skip sleep and throw together everything at once.
If there was anything Katara wanted to see the most at any point in the last three years, it was to see that Aang was in his best personal state while performing his duties. Back before the Day of Black Sun, she had been the first one to desperately get Aang to rest as he attempted to stay up for multiple days straight.
Thus, when she last saw Aang the night before, and found out of his current sleep drought, she quickly tried to get him to rest - alas, this was to no avail, as he quickly dismissed her and said that the work needed to be done as soon as possible, lest protests begin.
But this waterbender has always been determined.
That doesn't change.
Today - she vows that Aang will get a good night's sleep.
As night fell on the Southern Water Tribe, a bright moon illuminating a clear sky, Katara left her igloo and made the short walk to her boyfriend's home. Seeing the door closed, she assumed that he was continuing his paperwork. Still, she would not go home without the Avatar at rest.
Aang continued to fill out a response to a series of delegates from the Earth Kingdom, his eyes continuously closing as he fought off sleepiness. He was about three-quarters of the way through what he needed to do, and was hoping to finish it all, risking yet another lost night. He heard the knock at the door and turned his head. In the best voice he could, he muttered, "come in."
Katara opened the door. "Aang. How are you --" she stopped as she moved closer and saw Aang's tired look. She saw bags under his eyes, which kept threatening to shut themselves up for several hours. She also saw himself almost falling off the chair as his head drifted downward. "Oh my...Aang, how long has it been since you've slept!?"
"I took an hour-long nap about 12 hours ago," the Avatar mustered up.
"Just an hour?" The waterbender question, her voice dropping into a tone of worry. "Come on, let's get you out of this chair before you fall off it..."
Aang complied with this, getting out of the desk for what seemed like the first time in ages. What he had failed to mention was that he fell asleep right at the desk, which had given about a full day just sitting there in total. He knelt on the ground, Katara across from him, her hands on his shoulders.
"Aang, you look like you're about to fall onto this floor and sleep for about the next week," Katara said. "You really need to get some rest."
"But Katara," the Avatar began to protest. "I have to respond to those Earth Kingdom delegates, I have to tell the Fire Nation colonials about the pending republic, I have to also let Zuko know how well the conversations are going, establish future meetings with Kuei --"
Katara placed a finger over his lips. "Sweetie, it doesn't all have to be done tonight. I'm sure they'll all understand. You've been working tirelessly for the last three days and have gotten a lot done. You deserve to have a good night's sleep," she spoke calmly.
"Katara, I get what you're saying and all..." Aang mused, "but you know I have a duty to the world to make sure this all flows smoothly."
"That is true," the waterbender answered, "however, you also need to keep yourself in mind, Aang. You can't just go without sleep for days on end. If anything, not sleeping is making it worse. You're really tired and, after a while, you can't think straight. Remember before the Day of Black Sun? Or that time when we ran from Azula, Mai, and Ty Lee all night?"
Aang rubbed his eyes. "I suppose you're right, Katara. There's just so much to deal with..."
"You didn't have to master the elements in one day, did you?" Katara questioned him with a slight chuckle.
"Sometimes, it just felt like that..." the Avatar mused, thinking back to the fact that he had just nine months to at least get decent training or else the world would end in fire. "I don't want to let everyone down, though..."
Katara shook her head. "Stop worrying about letting everyone down. Getting sleep won't bring about the end of the world."
"I didn't tell you, Katara," Aang began. "During my nap before, I dreamed I had a full night of sleep, but when I woke up the next morning, I was getting word that the Earth Kingdom citizens were in full rebellion against the Fire Nation, and a new war was beginning. It was getting like the Battle for Yu Dao, but even worse...people were dying, their town was burning down...and I knew I had to get back to business or else that might happen."
Katara pulled him in for a hug, and dropped her voice to a whisper-quiet tone. "You know that won't happen just because you get some sleep. Everyone will understand, Aang. They aren't expecting you to solve anything overnight. This is why you have these meetings. Don't worry so much...you need to get rest...you and the world will be better for it. If you go much longer, it will really begin to hurt you..."
Aang nodded his head but quickly pulled it off Katara's shoulder, feeling himself drifting off. "I don't know..."
The waterbender cupped his right cheek in her hand. "Please, Aang..." she then smiled as she thought of something. "I'll tell you what...if you sleep, I promise that I will cuddle up with you."
"You'd do that?" Aang said, his looks brightening at the idea. "But, what about your brother or dad finding out?"
Only a light chuckle came from Katara. "Aang...I'm 17...and how many times have we cuddled up in the past? Sokka tells me he's surprised if I'm not laying next to you nowadays. So is Dad. As long as we're not doing anything else..." she thought back to stern warnings she had been given about her being with Aang. "...we'll be fine."
The airbender nodded, and the two got up. "Okay...I'll try to sleep."
Katara grinned, and the two made their way upstairs to his bedroom. Without hesitation, Aang fell onto the bed, right on his back. Katara came around to the right side, causing the Avatar to shift a bit to the left, allowing his girlfriend room. She placed herself in, throwing her arms around him, and lifting her head onto his shoulder.
Aang wrapped an arm around her back, trying to relax. Still, he murmured the words, "Earth...Fire...war...no...must help..."
"Not tonight," Katara whispered, kissing Aang on the cheek. "It's sleepy time. Take your rest. The world will not collapse...you've done your job the best and longest you can for now. Everybody needs to sleep...I'm here for you, Aang...sleep." She gripped him tightly.
"Katara..." Aang whispered back. "Guide me..."
The waterbender gently shut his eyes. "There. Now you can drift off. Don't worry about a thing...sweet dreams."
"I love you..." The airbender muttered as he rested his head on top of hers.
"I love you too," Katara replied with the same low voice. "I always will. You are my world, Aang...and tonight, I'm safe."
Perhaps that was the assurance he needed, because at that moment, Aang drifted off to sleep, smiling brightly once more...how Katara had missed that smile. She could only return it as she too drifted off to the world of dreams alongside him.
See more
For the collective works of the author, go here.


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Knots: Hitches
Anchors aweigh! This framed mini print is your how-to to a host of Hitch knots, helping you rig a rope to an assortment of objects! Illustrating everything from the Clove to the Pruisk to the Midshipman's Tautline, rest assured, these hitches be crazy (cool).
Available individually, or as part of our Knot Set!
7" x 5"
Each mini print is pressed on 100lb archival stock and comes in a baltic birch black frame manufactured in our Brooklyn wood shop.
Please note that due to its custom assembly, this product requires an additional 5-7 business days of processing time.


text: Most Soul Calibur games can be used with a cheat device called Action Replay. The most notable use of an action replay is being able to use inferno in Soul Calibur II. And Soul Calibur 2 has it's very own cheat device titled Soul Calibur II:Ultimate Codes. Besides characters, action replay discs allow to get infinite money,health,etc.


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Listing Courtesy of Bill Doran of Elcor Realty of Rochester, Inc.
Listing information
- Beds: 2
- Stories: 0
- New Construction: No
- Above Ground Sq. Feet: 1,283
- County: Olmsted
- Elementary School: Ben Franklin,Pinewood
- Style: Side By Side
- Baths: 2 (Full: 1 3/4: 1 1/2: 0 Other: 0)
- Fireplaces: 1
- Foreclosure: No
- Estimated Annual Taxes: $516
- High School: Mayo
- School District: Rochester #535
- Year Built: 2015
- Status: Sold on 8/31/2015
- Parking Type Description: Attached 3 cars
- Total Square Footage: 1,283 ($162.43 per sq. foot)
- Association Fee: $55
- Middle School: Willow Creek
- Subdivision: Wildwood Meadow
3 Bdrms on One Flr No
Air Conditioning Central
Amenities - Exterior Driveway - Concrete, Garage Door Opener, Patio
Amenities - Interior Cable, Ceiling Fan(s), Kitchen Center Islnd, Kitchen Vent, Kitchen Window, Master Bathroom, Master BR W/I Closet, Pantry, Vaulted Ceilings, Walk-in Closet
Appliances Dishwasher, Microwave, Range/Stove, Refrigerator
Association Fee Incl Lawn Care, Snow Removal
Association Fees: Assn Fee Frequency Monthly
Association Fees: Planned Unit Dvlpmnt No
Basement Features N/A
Basement Material N/A
Basement Style None
Bath Description 3/4 Master, Main Floor Full Bath
Construction Type Frame/Wood
Dining Combine with Kitchen
Exterior Stone, Vinyl
Financing Conventional
Fireplace Yes
Fireplace Type Gas
Fuel Natural Gas
Garage Type Attached
Heat Fireplace, Forced Air
Mechanicals Water Heater - Gas
Miscellaneous: DPR Eligible No
Other Finished Above Grade - 1283.00, Finished Below Grade - 0.00, Garage Type: Garage SqFt - 640, Square Feet: Above Ground SqFt - 1283, Square Feet: Below Ground SqFt - 0, Square Feet: Main Floor SqFt - 1283, Square Feet: Total SqFt - 1283, Tax Year - 2015
Rental License No
Roof Asphalt Shingles
Sewer City
Special Search Main Floor Bedrooms
Stories 1 Story
Water City
Waterfront No
Waterfront Name None
Bathroom M x
Bedroom 2 M x
Dining Room M 10x12
Kitchen M 16x9
Laundry Room M x
Living Room M 16x15
Master Bedroom M 13x14.5
Sun Room M 12x12
Listing Courtesy of Bill Doran of Elcor Realty of Rochester, Inc.


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Anniversary Sale
DL1961 Florence Instasculpt Skinny Jeans (Hail)
Sale: $99.90
After Sale: $178.00
AG The Legging Ankle Super Skinny Jeans
Bold whiskering creates rich highs and lows over the svelte silhouette of super-skinny jeans cut to a versatile ankle-grazing length. Color(s): 04 years rapid, 17 years sea drift. Brand: AG. Style Name:Ag The Legging Ankle Super Skinny Jeans (04 Years Rapid). Style Number: 5389564. Available in stores. Learn More
DL1961 Farrow High Waist Instaslim Skinny Jeans
A sophisticated, fade-resistant black wash adds day-to-night versatility to skinny jeans made with a leg-lengthening high waist. This wardrobe staple lifts and sculpts your figure with DL1961's Instaslim four-way stretch denim. Color(s): hail. Brand: DL 1961. Style Name:Dl1961 Farrow High Waist Instaslim Skinny Jeans (Hail). Style Number: 5182450. Available in ... Learn More
DL1961 Florence Instasculpt Skinny Jeans
Figure-hugging skinny jeans instantly sculpt the waist, bottom and legs while providing comfort and shape retention. Subtle whiskering and fading add dimension and lived-in authenticity to this versatile style. Color(s): carry-on. Brand: DL 1961. Style Name:Dl1961 Florence Instasculpt Skinny Jeans (Carry-On). Style Number: 5391285. Available in stores. Learn More
DL1961 Ryan High Waist Skinny Jeans
Slim from the waist to the ankle, these figure-defining high-rise jeans are designed to fit and flatter a petite frame. This weekend-ready pair is cut from cotton-blend denim and distressed with allover sanding, threadbare patches and crisp 3D creases. Color(s): stingrey. Brand: DL 1961. Style Name:Dl1961 Ryan High Waist Skinny ... Learn More
FRAME Distressed Release Hem Skinny Jeans
London-inspired style combined with the quality of the LA denim industry defines fresh go-to jeans styled with trend-right released hems, edgy rips at the knees, and supersoft and stretchy tech denim that offers a flattering fit. Color(s): lilac blanc, lilac film noir. Brand: FRAME DENIM. Style Name:Frame Distressed Release Hem ... Learn More
FRAME Le Skinny de Jeanne Raw Hem Skinny
London-inspired style combined with the quality of the LA denim industry informs a chic pair of go-to jeans updated in a faded grey wash and finished with two tiers of shredded denim at the hem. Supersoft and super-stretchy tech denim offers a flattering fit that slims the thighs while lifting ... Learn More
FRAME 'Le Skinny' Sateen Skinny Jeans
Soft whiskering and subtle fading detail super-skinny brushed stretch-sateen jeans that combine the iconic edge of London style with LA's innovative denim heritage. Color(s): knightrider. Brand: FRAME DENIM. Style Name:Frame 'Le Skinny' Sateen Skinny Jeans (Knightrider). Style Number: 1012032_3. Available in stores. Learn More
HUDSON JEANS Barbara High Waist Super Skinny Jeans
A high waist and dark blue wash add instant sophistication to super-skinny jeans cut from delightfully soft denim that recovers wonderfully for the perfect fit that doesn't sag. The shadow of Hudson's signature button-flap back pockets finishes the rear of this playful style. Color(s): calvary, recruit 2. Brand: HUDSON. Style ... Learn More


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Burning scalp syndrome is scientifically known as trichodynia or scalp dysethesia. It commonly affects people suffering from hair loss conditions, like androgenetic alopecia and telegon effluvium. Burning scalp syndrome is characterized by painful burning sensations in the scalp and around the roots of hair. Although a clinical cause has not been determined, scientists believe that "substance P," a neuropeptide made by nerve cells, may be to blame. Burning scalp syndrome can be incredibly painful, and while there is no perfected cure, dermatologists have had success treating the condition with antidepressants.
People suffering from burning scalp syndrome complain of burning, itching and stinging of the scalp. These symptoms are often triggered or made worse by physical or psychological stress. Trichodynia, unlike scalp dysethesia, is also characterized by painful sensations in the hair itself, which become more intense when the hair is touched.
According to "Clinical Management in Psychodermatology," 34 percent of people suffering from chronic hair loss syndromes experience burning scalp syndrome. The pain occurs with no physical pathological findings, and is thought to be a somatoform disorder. Somatoform disorders are psychological disorders in which physical symptoms appear or are felt with no predictable or understandable medical cause.
According to keratin.com, doctors do not precisely understand the cause of burning scalp disorder. However, research has shown that neuropeptides in the skin and hair follicles may be involved. Substance P fluctuates in amount as hair follicles grow and die. It has been shown to create sensations of pain, and is thought to become more prevalent as hair follicles die. Another explanation for burning scalp disorder is that cell activity caused by androgenetic alopecia or telegon effluvium produces inflammation, which results in pain. There could also be underlying psychological problems that cause the disorder.
According to "Clinical Management in Psychodermatology," just over three-fourths of patients with burning scalp syndrome also have psychiatric disorders, such as depression, anxiety disorders or obsessive-compulsive disorders. Burning scalp syndrome has also been linked to fear of hair loss, and this fear may produce painful sensations in the scalp.
According to keratin.com, burning scalp syndrome is sometimes treated with tricyclic antidepressants. While usually taken to relieve depression, tricyclic antidepressants have been proven to help reduce levels of substance P. It is thought that reduction of substance P levels will help ease sensations of pain. Other treatments for burning scalp syndrome include corticosteroids, capsaicin, anesthetic agents and gentle shampooing.
Related Searches
- Burning Scalp Syndrome
- Harth, Wolfgag and Gieler, Uwe. Clinical Management in Psychodermatology. Springer-Verlag, Berlin: 2009.
- Scalp Dysesthesia
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John (Jack) McLean survived 32 missions fighting the Nazis as a gunner in a heavy bomber and now, almost a lifetime later, it seems, Canada is awarding veterans such as him a special distinction to mark their service.
The Harper government took the wraps off a commemorative bar in Ottawa Monday to honour the 50,000 Canadian veterans who served in Bomber Command. Of the 55,000 Allied men and women who lost their lives in the bomber forces, more than 10,000 were Canadian.
As the mid-under gunner in his Halifax bomber, Mr. McLean had a bird's eye view of the devastation of war.
"I was looking right down at the target: the explosions, the fires. It was just pandemonium and it's something you don't forget," said the veteran, who served as an 18- and 19-year-old and is now 87.
"So many nights, Allied air crews flew into the dark, cold and lonely skies over occupied Europe, knowing their odds of surviving their tour were barely 50-50," Veterans Affairs Minister Steven Blaney said Monday as he unveiled the bar with veterans such as Mr. McLean in attendance. "They had every reason, every right to fear that their next mission might be their last. And yet they answered the call to duty night after night, day after day."
Mr. Blaney acknowledged the long delay in recognizing Bomber Command vets. Canada, he said, was offering vets a "thank you the way we always should have done."
The Allied bombing raids were controversial because of the large number of German civilians killed by the attacks. "For long stretches of time, the air was the main and almost only weapon we had to fight the occupying forces on the European continent," the minister said.
Mr. McLean said his aircraft was "one of the lucky ones." It caught lots of flak during the six- to seven-hour missions, but never sustaining a major hit as it carried up to seven tonnes of bombs and incendiary devices to targets mostly in Germany's heavily industrialized Ruhr Valley. "The flak was very heavy. The Germans had their night fighters. They had good aircraft but they didn't get us. We had a good pilot."
The constant noise in the aircraft left its mark on him. "That's why I have hearing problems," Mr. McLean said.
He lost two good friends – both bomb aimers – who were killed in training exercises in Scotland and England. They were on his mind during the ceremony Monday.
Mr. McLean, who lives in the Ottawa area, has no regrets about his wartime experience. "I've never been sorry I had it," he said, adding his best memories are the camaraderie. "We were young Canadians, flying together and taking our chances together."
Only two others of his 11-member bomber crew are still alive.
The veteran says he's grateful for the honour, despite the wait. "It took around 70 years to get this, but it's nice to get it in the end."
Of the 50,000 Canadians who served with the Royal Canadian Air Force and Royal Air Force in Bomber Command, relatively few are still drawing breath today. The government is not offering official estimates but expects perhaps less than 2,000 veterans remain.
The new commemorative bar will be offered to veterans, or their surviving spouses or "loved ones" of deceased veterans, the government said. The only criterion is one day of service in Bomber Command, either in the air or on the ground as support crew.
Mr. McLean says the scariest part of the mission for him was takeoff for the heavily loaded bomber. "Anything could happen and once it did, down you came."
He says the best part of the mission – during which he wore electricall –heated suits to brave the cold at 20,000 feet – was landing safely.
"It was always good to get back and have a cigarette, a cup of coffee and be debriefed before going to bed, getting ready for the next one," Mr. McLean recalled.Report Typo/Error
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Blondieee Model
Thomas DunkerleyPhotographer Male Bellevue, Washington, US
My Website: Crystal Light Photography Home PageMy MM URL: http://www.modelmayhem.com/CrystalLight
Mayhem # 483
About Me
Greetings! My name is Thomas Dunkerley (the other one). I have been actively involved in the business of glamour photography for the past twelve years. Currently I am concentrating on working with new and experienced models alike by adding quality imagery to their modeling portfolios. I am available for anything ranging from high-end fashion, glamour, artistic nude and the erotica. I pour my heart and soul into my work and strive to produce only the finest quality imagery for models that wish to work with me. And while I can provide all of the imagination and creativity for a shoot, I also love to listen to and work with my models for their ideas as well.
While of course I prefer paid opportunities, I am open to very generous and timely TFP/CD with select models that I feel will help promote my work. Don't hesitate to ask!
I have but one current personal project on my plate, and that is the creation of artistic glamour photo books. I have completed my first one (see below) and am now starting my second book.
About me: I love art, beauty and women; thus my focus on the world of glamour. I am a very high spirited individual that loves to laugh and have fun. I am married to the most loving and supportive wife imaginable.
If what I have wrote above interests you and you are desiring to work towards building your portfolio in a relaxing and non-intimidating manner, please feel free to contact me. Thank you for your time!
Have you worked with Thomas Dunkerley? Add Credits for Thomas Dunkerley! >>
Jessica Converse
See Sample Worked together 2-10 times; Most recent: August 2012
Credit Notes
I've just recently published a 240 page photo book entitiled Undiscovered Beauty, ISBN 978-1-4276-3477-1, registered under Aardvark Global Publishing, featuring 50 lovely models in various settings and stages of dress and undress. You can preview and purchase Undiscovered Beauty at: http://www.blurb.com/bookstore/detail/454297 . Special thanks to all those models that participated in this project!
Blondieee Model
Maegan Model
Boyana Marinova Model
Kara Jean Model
Red Scorpio Photographer
LouAnn Eads Model
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text: I'lll start out our fairy tale by saying how we met. I started working at a daycare and everyone there was trying to get me to date someone they knew that was single. I finally said yes to one girl and so she set up sort of a blind date with her boyfriend's friend. I got to see a small picture of him and he got to see a small picture of me, but it was hard to tell what each one really looked like, but we both agreed to meet. Well we met and hit it off. Eleven months after meeting my fiancé finally popped the question! Christmas Eve he told me we were going to see Christmas lights. We left a Christmas get together and proceeded to the lights. He then told me he forgot the tickets so we would have to stop by his house. We pulled in the driveway and I asked if I should stay in the truck. He told me I could come in for a minute. We went inside and he asked his sister where he put the tickets. She said you were reading a book and you put them in that out on the porch. I started thinking why was he reading a book outside in the cold? He turned the porch light on then off and went outside. I started thinking again why did he turn the light off when it's dark out on the porch and he is looking for a book? He then asked "Are you going to help me?" I said "I guess." So I went out to help and he closed the door then hugged me from behind. He then started telling me that he read in Genesis that day. I said "oh yeah?." He said "yeah I was reading for inspiration. You know how God told Adam he would give him a help mate?" I said "yeah." He let go, came in front of me, got down on one knee, started pulling a box out of his pocket and said "well God gave me you as my help mate." Then he turned and I asked "what?" I turned and looked over the banister at a sign that he made with Christmas lights (that I later found out I helped pick out.) that said "Marry Me" I said "aww yes!" Then he asked "Will you Marry Me?" I said "yes" and his sister came out taking pictures like paparazzi. Later I asked as I looked at my ring, in front of his while family, "Is this the only present you were talking about that I get to open tonight?" He laughed and I said "Oh no! That's not what I meant! I meant is this the present you were talking about?" Still didn't sound that great, but I meant the best even though I was really embarrassed!! If we were to win the prize we would put it in savings for our wedding since we are paying for our own wedding or for part of a down payment on our first house together.


text: Marco Polo Canada West: The Travel Guide with Insider Tips This compact, straightforward guide is clearly structured for ease of use. It gets you right to the heart of the region, and provides you with all the latest information and lots of Insider Tips for a thrilling Canadian adventure.Includes a road atlas and an additional pull-out map. Clear, user-friendly structure and layout. Get your bearings with the 'Where to Start' panels and ensure you don't miss out on the key sights using the 'Highlights' section. The 'Best Of' pages feature unique aspects of the region and also suggest places to go for free, tips for things to do when it's raining and good places to relax.Insider Tips and much more besides: Marco read more


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Kids Bowl Free provided a free family pass in participation with this article, all opinions 100% my own.
Now that summer vacation is in full swing, families are looking for entertaining activities to keep the kids busy throughout the summer. Since many are on a budget, finding free or low cost activities can be a challenge. But thanks to Kids Bowl Free, kids can enjoy the very popular sport of bowling.
Kids Bowl Free is a free summer bowling program allowing children to bowl two free games of bowling every day all summer long! And what is great about the program, is up to six registered children can play. Kids Bowl Free is amazing for small or large families so children can have something to do every single day of the summer…FOR FREE.
Bowling is a fun sport even toddlers can play. I started bowling at age 4, though I am no longer on a league, we still enjoy bringing the kids to play. In an enjoyable way, children will also get exercise burning calories without knowing it. Just think, the kids may enjoy bowling so much, they'll want to play this new sport and join in on their own league. Bowling also makes a wonderful birthday party idea.
Registration is simple, by adding parents information and registering up to six children, you'll be signed up and coupons will be emailed to print to take bowling.
Parents if you are interested in bowling with your child during the summer, there are family passes available for $24.95 for the adults. Family passes are good for up to four adult family members. In my experience, bowling costs about $3 per game, so this is a wonderful deal to be able to bowl daily with your children. I know we are taking advantage and going bowling every single day!
To learn more about this program and to get your children registered, visit www.kidsbowlfree.com and check to see if a bowling center near you is participating!
Stay connected with Kids Bowl Free on Facebook and on Twitter @kidsbowlfree


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Call Her Fawn Wielichowski, Sister Says, Since Husband Nikolas Gianes Killed Her
A Facebook photo of Fawn Wielichowski and her husband, Nikolas Gianes, who's accused of killing her. Additional photos below.
This morning, the Colorado Springs Police Department formally identified the woman who died in an October 16 homicide as 32-year-old Fawn Christine Gianes.
However, the victim's sister, Krystal Wielichowski, asked KRDO-TV in the Springs to refer to her by her maiden name and put the word "Gianes" in parentheses.
"She's a Wielichowski," Krystal told the station from her home in Missouri, adding, "That man murdered her,"
"That man" is Nikoas Gianes, Fawn's husband. He's been arrested on suspicion of second-degree murder.
KVOR.com identifies Nikoas as a onetime El Paso County sheriff's deputy, and that seems to be confirmed by his Linkedin page, which also references a military background, as seen in the following screen capture.
There are plenty of other images of Nikoas and Fawn on her Facebook page, which remains online at this writing.
Here's one example.....
...as well as a shot of the couple with their family, which we're sharing since the children are seen from the back, thereby disguising their identities.
Unfortunately, such idyllic imagery appears to have hidden some dark secrets.
"There has been suspicion of abuse, there have been prior incidents, but none to this extent," Krystal told KRDO, adding that when she heard the news about the shooting, "it was numbing, I thought it was a cruel joke. I didn't know what was happening. I hit the floor."
What took place?
At around 7:31 p.m. on Friday the 16th, according to the CSPD, officers were dispatched to a home on the 5300 block of Babcock Place on a report of shots fired.
Upon their arrival, they found a woman later identified as Fawn. She was dead from an apparent gunshot wound.
A Facebook portrait of Fawn Wielichowski.
Nikoas was taken into custody shortly thereafter.
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He is being held without bond in the El Paso County Criminal Justice Center.
In the meantime, a GoFundMe page has been launched under the title "Bring Fawn Wielichowski Home." The introduction reads:
Fawn Christine Wielichowski was a loving mother of 2 beautiful children. She brought great joy to many of us with her fun loving attitude and her ability to make us laugh. She was tragically taken from us in a way that no one ever should be. She will be missed by her family and friends and her memory will live on within us. Please help us bring her home.
Any donations are greatly appreciated. Donations would be used for creamation and funeral cost. Travel to CO and back. Any extra funds will be used to assist her children with living arrangments.
We are so thankful for everything!
At this writing, just over $1,500 has been pledged toward a goal of $2,500. For more information, click here.
In the meantime, our sincere condolences to the friends, family and loved ones of Fawn Wielichowski.
Look below to see Nikoas Gianes's booking photo.
Nikolas Gianes.
Colorado Springs Police Department
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text:
Welcome from our Head of School
I'm delighted to introduce you to K12 International Academy—a fully accredited, U.S. diploma-granting, private online school for grades K–12. All around the world, we help students learn in the ways that are right for them, nurturing a joy for learning and a passion to pursue their interests.
The key is our individualized approach, which frees children to reach their personal potential. Our students are liberated from rigid schedules, classes that move too fast or too slow, bullying, overcrowded classrooms, teaching to the test, and many other factors that typically stand in the way of success.
It all starts with the award-winning curriculum from K12, America's K–12 online education leader, which was developed by cognitive scientists and master educators based on decades of research into how children learn. So every lesson comes alive with interactive content and hands-on materials that keep kids engaged.
We also surround you and your child with an amazing support team, including online teachers who have a true gift for inspiring young learners. Working with our academic leadership, guidance counselors, and other education specialists, they'll be with your family every step of the way.
And you'll reap the advantages of our energetic school community—an international network of friends who assist, encourage, and learn from each other through online student clubs, parent roundtables, and much more.
If you're looking for a school that can adapt to your child's needs and rekindle the joy of learning, I invite you to learn more on the following pages. Join us full time, or even for a single course, and the rewards will be worth it—from the newfound flexibility and focus, to all those 'a-HA!' moments to come.
Miriam Rube
Head of School, K12 International Academy
Miriam Rube has been an educator for more than 25 years, with experience both in the classroom and as a principal. She received her MS in guidance and counseling from St. Thomas University in Miami, Florida, where she is now a doctoral candidate in Instructional Leadership and Curriculum. She is also currently a member of the Principals' Center at the Harvard Graduate School of Education, following a three-year term on the Harvard Advisory Council.


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Bright Light Precure! (明るい光プリキュア! Akarui Hikari Purikyua!) is Cure48's 11th fan series.
Bright Light Precure! Episodes
The story begin a one years after Light Pretty Cure, Hikari town is in danger, again a power of dark is attacked Hikari town ( a town where the cures live ). They have an enemy again Darkness Cave, and they must save human dream from Darkness Cave.
Pretty Cure
Shibuki Hikari
Voice Actor: Mizuki Nana
Hikari is the leader of Light Precure. Hikari is a famous idol, she always dream about a girl fighting with many monster. Later, she become Cure Sparkle in episode 2. Her alter ego is Cure sparkle. She Introduction: "The Everyone Light, Paceful Future! Cure Sparkle!"(みんなの光平和の未来!キュアスパークル! / Min'na no Hikari, Heiwa no Mirai! Kyua Supākuru!).
Mizuki Aoi
Voice Actor: kanae ito
Aoi is the sporty girl of the team. She is very sporty and is best friends with Hikari. Her alter ego is Cure Grace (キュアグレース/Kyua guresu). She becomes Cure Grace at episode 3, and she Introduction: "The Beautiful Swan on the Lake! Cure Grace!"(湖に美しいの白鳥!キュアグレース! / Mizūmi ni utsukushī no hakuchō! Kyua Gurēsu!)
Fukuhara An
Voice Actor: Kugimiya Rie
An is the senior girl of the team. She is also an idol, but she in Hikari rival agency. She become cure happy before the story begin. Her name is same as character of pretty rhythm rainbow live but different face, her ego is cure happy same as Miyuki, but different introduction and dress. Her intoduction is " everyone smile, cure happy! ".
Watanabe Naru
Voice Actor: kana Asumi
She is a new student at Hikari, Aoi, and An class. She is shy and crybaby, she become Cure Sapphire in episode 4 and her introduction is "The green stone, cure sapphire! "
Town Of Light
Rock is a main mascot in this fan series, she has a friend named sweet.
She is a friend of rock, she help rock for find pretty cure and in second season she active to help Rock and teh cures.
Lucky Queen
The queen of , town of light.
Darkness Cave
Kurohime - Kurohime is the first enemy to attack the Cures. She is very mature girl, she also yuri. She is the only villain who doesn't die.
KurohYume - she strong and she never forget people who hurt her, she is tomboy and she is very feminine, she always bring a black umbrella.
Joker- The last enemy , Mask is a only boy enemy.
Darkness King - The King Of Darkness Cave.
Memory Catch - The transformation device.
Magic Mirror- is a attack device, when attacked a hug Lucky Queen appear, just like Heartcatch Precure attack.


text:
Emerging Trends in Tourism and Travels
Do you ever wonder about how you can shape your future in Tourism and travel industry when you contemplate your career in tourism management? The hospitality management courses include some new astonishing trends in Tourism and travel:
The New Concept of New-Style Lobbies:
Traditionally, there has been a transitional zone in hotel lobby, as these travelers used to check-in there is a place for them to park their baggage. But this concept has changed to a huge extent; now lobbies are considered as a potential place for gathering by the tourism management staff, where the guests would never want to leave such lobbies and if desired they can even get to know each other. Curate bookshelves, espresso machines, and coffeehouse tables are the main features and elements of these new style lobbies.
Bid Adieu to Business Class:
According to a recent survey done it was found that over the coming year, 58% of journeyers are expected to travel less for the purpose of business. The orientation of the future strategies of tourism management inclined towards the under-25 market for which the significance is increasing rapidly.
Practicing Online Booking:
The times when you use to call from your land-line for making a booking in hotel appear to be over. Today the stays are booked online by most of the guest. The overview of the latest Internet Booking Technology must be included in the Hospitality Management.
Boutique Hotel evolution:
As the travelers opt for the smaller ones, thus the era of the big-name hotels may be over, instead of this boutique options can be personalized more.
Bio-metric Check-in:
Unlike the airports and various other places soon the hotel lobbies will have the feature of Self check-in kiosks. This feature is appealing especially for those travelers who would like to check in instantaneously and quickly. These features also help in saving the expenditures on Human resources.
Personal Care Products:
Pumps are being installed in several hotels for the personal care products in the shower. The double advantage of saving the money can be taken by these devices as we can reduce the cost of packaging that is done on the complimentary mini bottles of shampoo, lotion and conditioners.
Showers rather than bath:
Just to satisfy the special needs of customers or who are traveling with small children for them keeping a small percentage of baths on one hand, the hotels nowadays are building more bathrooms with showers.
Hope you are intrigued by these trends of Hospitality management and will implement these, if you are or if you wish to make your career in Tourism's and travels.


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The R&D Management Conference 2012
Creating and capturing value through R&D management and innovation
Grenoble, France 23-25 May
Conference theme
The aim of The R&D Management Conference 2012 is to comprehensively explore how 21st century organizations will create, capture and deliver value. Papers are welcomed from a broad range of scientific communities and industrial sectors. They should illuminate the implications and opportunities and advance the theory or practice of R&D and innovation management.


text:
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When you purchase DRM-free downloadable content, that content is available for you to download on any computer and save to a disk whenever you like. You can also download to multiple computers. There is no timeframe associated with your purchase and the content will not expire.
Running Time: 82 minutes
Released: 03/2010
Studio Name: Candy Shop
Director: Phil E. Blunt
Categories: Black Natural Breasts All Sex
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This is a Rookwood Vase with a Pansy design. It does have some grazing and there are some rough spots along the rim. It's still a beautiful collectible Rookwood piece. Measures 3 1/2" high and 4" wide.
A Good Selection Of Yesterdays Treasures
Welcome to Cameo Antiques, here you will find high quality Antiques and Collectibles at affordable prices.


text:
Valumark Pizza Cutter with Bottle OpenerSKU #: LM-VR3103
Item DescriptionMore InformationAdditional Information
A complete tool-in-one for pizza night, this Valumark Pizza Cutter with Bottle Opener is also great to use for quesadillas, cookies and bars, pies, quiches, and more. This pizza cutter is made of strong ABS plastic, measures approximately 7-3/8" W x 3-1/2" H x 7/16" D and features a hanging hole to easily store away. The bottle opener side can be used for beer, wine coolers, root beer, and more. It's available in white or red and can be customized with an imprint such as a logo or company name to use in promotions, tradeshows, and more. It would also make a great item for giveaways, company events like seminars and conferences, or give to employees at company holiday parties.
Price includes the cost of a one color imprint in one location. Please contact us at 888-206-3047 for more print options, a custom project, or additional print locations.
|24 Hour Rush
|ABS Plastic, Steel
|Product Dimensions
|7 3/8 W x 3 1/2 H x 7/16 D
|BPA Free
|BPA Free
|Dishwasher Safe
|Not Dishwasher Safe
|Microwave Safe
|Not Microwave Safe
|Imprint Method
|Pad Print
|Imprint Dimensions
|1 1/2'' x 9/16''
|Maximum Number of Colors
|Standard Production Time
|5-7 business days
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text:
John Holmes ExposedFavorites [+]
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Running Time: 81 minutes
Released: 01/1995
Studio Name: Gourmet Video Collection
Description: The most amazing, biggest man in porn, John Holmes and his amazing big thick 13 inch cock gets to satisfy 15 of the hottest nastiest girls in porn. Whether he does them individually or 2, 3 at a time, they love to suck his massive member and get they're pussies stuffed hard and deep and lap up all his cum!
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This is a disambiguation page; that is, one that points to other pages that might otherwise have the same name. If you followed a link here, you might want to go back and fix that link to point to the appropriate specific page.
Annunaki (also transcibed as Annunaku) can refer to several things. Did you mean:
- Annunaki, primal entities revered among the ancient babylonians in Mage: The Ascension?
- Annunaki, also known as Malefactors, the third House of the Angels in Demon: The Fallen?
- Annunaku, a Gangrel bloodline in Vampire: The Requiem?
- Annunaki, abyssal beings of near divine power in Mage: The Awakening?


text: McCall's 9121; ©1984; Misses Skirt, Straps and Bibs: Dirndl skirt A, B or C is gathered to buttoned waistband, has back zipper opening and large patch pockets; waistband has front and back buttons for detachable straps or bibs with buttonholes. View A skirt has straps. View B has self-lined bib with patch pocket and shoulder straps. View C has lined front and back bib


text:
Cantera: An Object-Oriented Software Toolkit for Chemical Kinetics, Thermodynamics, and Transport Processes. Cantera is a suite of object-oriented software tools for problems involving chemical kinetics, thermodynamics, and/or transport processes. Cantera provides types (or classes) of objects representing phases of matter, interfaces between these phases, reaction managers, time-dependent reactor networks, and steady one-dimensional reacting flows. Cantera is currently used for applications including combustion, detonations, electrochemical energy conversion and storage, fuel cells, batteries, aqueous electrolyte solutions, plasmas, and thin film deposition.
Keywords for this software
References in zbMATH (referenced in 3 articles )
Showing results 1 to 3 of 3.
- Hansen, M.A.; Sutherland, J.C.: Pseudotransient continuation for combustion simulation with detailed reaction mechanisms (2016)
- Yonkee, Nathan; Sutherland, James C.: PoKiTT: exposing task and data parallelism on heterogeneous architectures for detailed chemical kinetics, transport, and thermodynamics calculations (2016)
- Lv, Yu; Ihme, Matthias: Discontinuous Galerkin method for multicomponent chemically reacting flows and combustion (2014)


text:
We're shifting from an ownership econcomy to an access based economy. As more and more media moves into the "cloud", as devices like smartphones take over the roles of more and more discreet devices, and as on demand 3D printer looms on the horizon, Stan & Jason ponder discuss how we're at the beginning of an era of "less stuff". How might this shift away from physical goods affect the environment? An upgrade based economy? Our identities? JOBS?
**Note** Apologies for Jason's robot voice in the last few minutes of the episode. The gremlins in his headset seem to get upset after they've been recording his voice too long **


text:
AS part of the Big Brownie Birthday, celebrating 100 years of the organisation for youngsters, members from 2nd Studley (St. Mary's) Brownies blasted off into space with 150 other girls from across the county on a sleepover at the Space Centre Leicestershire.
]Four events like this are taking place on weekends to enable 600 girls to sleep under the stars and experience space.
Brownies made flying rockets, presented a TV weather report, viewed the night sky up close and took a 3D trip into space.
Group leader Caroline Ford said: "Today's Brownies expand their horizons and there are no limits to girls achieving their full potential within Girlguiding."
For more details, visit www.girlguiding.org/interested.


text:
Randoms at Wikia
The beta testing for the board game for version 1 was held with the creator and to sub-creators. The prize for entering the beta test was the beta crystal whitch was chosen by random. If you won the tournament you got a skill point plus 100 star bits.
|This article is a stub. You can help Scratchpad by expanding it.


text:
Read How Obama Wants to Steal MONEY from Every American!
This must be stopped!Typical Obama stuff!President Trump has announced that he does not want a salary for being the President,while Obama is trying to get himself a raise before being terminated!Congress do your job and and stop Barack Obama before he destroys us more!
According to ConservativeTribune Barack Obama continues with his liberal agenda, although there are only two months to the end of his term.
American Action Forum reported that the 10-year regulatory burden on Americans has now passed $1 trillion.
Over $1 trilion are spent on trucks for New Environmental Protection Agency..
That means now every American now will have to fork over $3,080 at once for these new regulations, or an annual cost of about $540..
Obama, thank you!
High taxes and few jobs can easily get to these payments. These negatives associated with the corruption of the federal government.
Thanks to all these new regulations, the estimated number of hours for all the paperwork is 754,208,700 hours. As all that was not bad enough.
It takes about 86,097 years for a person to complete, or will be required 377,000 people working full-time at 2,000 hours a week to process. For the US economy none of their work аdd any value.
Obama before leaving office, probably will not try to push through more regulations.
Can we make the lives of Americans even more miserable before Obama moves out of the White House? "Yes, we can!"
Obama before leaving the White House can the lives of Americans to be more miserable? "Yes, of course!"
Trump announced on his website during the campaign that federal regulations will be drastically reduced after we entered the White House.
Obama and his administration have spent so much money on other people, besides Americans, and expect from us to make more money to give to the government so they can pass out more.America will definitely be Greater when this Obamanation is finally gone!
What do you think about this article?? Share your thoughts in the comments below.


text:
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Last seen: January 18, 2013
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ü Aircraft Structural Design
ü Automotive Product Design
ü CAD Migration
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ü PDM- Enovia LCA, and SAP
ü Project management
ü Operating Systems: MS-DOS, UNIX,Window2000/NT/XP
ü Excellent Communication and presenting Skills
ü Product Development, Design, Packaging Study, CAD Conversion
ü 8+ years of Robust experience in CATIA V5.
ü Experience in working Domestic and Oversea Customers
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text:
Product Description
Butler Vintage Oak Console Table 2130001
Selected solid woods, wood products and choice veneers. Cherry veneer top, sides, drawer front and lower display shelf. Drawer with resin applique and antique brass finished hardware.
- Sku: 2130001
- Manufacturer: Butler Speciality Company
- Collection: Plantation Cherry
- Category: Consoles/Sofa Tables
- Finish: Vintage Oak
- Finish Distressing: Light
- Dimensions: 37"W, 15-3/4"D, 34"H
- Carton Dimensions 1: 41W x 22.5D x 12.5H, Weight: 80, Cubes: 6.68
- Dimensional Weight: 70
- Shipping Method: LTL
- K.D. or Set up: K.D.
- Country of Origin: China
- Minimum Order/ Pack Quantity: 1
- Number of Shipping Cartons: 1
- UPC Code: 797379010029
Additional Information
Additional Information
|Free Shipping
|Delivery Type
|LTL Curbside Delivery with Appointment
|Delivery Time
|2-4 Weeks
|Butler Speciality Company
|Small Parcel
Product Tags
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text:
70 Minute DVD Program
Few foods create as much controversy as nuts. We often hear that they nuts are good sources of many nutrients including fiber, protein, and healthy fats. Nuts are also said to lower our risks for heart disease, diabetes, and stroke, and maybe even help to manage our weight. Can that be true, with all the fat and calories in nuts?
There is so much conflicting information out there, it's no wonder that people are confused and literally going nuts over nuts! Fortunately, in this easy to understand and entertaining 70-minute talk, Jeff Novick will reveal what the science really says about nuts and health.
Jeff Novick is truly a unique dietitian and nutritionist. His insightful and humorous approach to nutrition and health has helped thousands worldwide make the transition to healthy living. He holds both undergraduate and graduate degrees from Indiana State University in nutrition with minors in Exercise Science.
Jeff has taught nutrition classes at Indiana State University, Indiana University Medical School, the University of Miami Medical School and for the Florida Academy of Family Physicians and lectures at medical conferences around the country on a regular basis. While in indiana, he created and taught the Nutrition Education Initiative, a preventive medicine curriculum for medical doctors, residents, and medical students. In recognition of this groundbreaking project, Indiana's governor awarded him the Indiana State Public Health Excellence in Health Science Award, and Indiana State awarded him the Graduate-of-the-Last-Decade Award.
Shipping/Replacement/Returns: Shipping inside the US is included in the price. When your order is shipped, you will receive a notification with your tracking number. We will replace damaged or defective materials, or incorrect items shipped. No DVD returns accepted, all sales final.
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PRICE INCLUDES SHIPPING INSIDE US 90 Minute DVD Program ABOUT THIS PROGRAM Obesity is a...
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