text:
Prairie View A&M University has partnered with GradGuard to offer PVAMU students withdrawal insurance in the event of an unforeseen withdrawal due to medical or health related issues. Each year thousands of college students are unable to complete classes for the semester due to unforeseen medical issues such as:
- Illness,
- Accidents,
- Mental health related issues, or
- Death
Unfortunately, in many cases these families are unable to recoup all of their tuition dollars. While attending a university in the state of Texas and depending on the timing of the withdrawal, you may be responsible for paying all or a portion of the tuition and fee charges and may not receive a 100% refund according to the Student Withdrawal Refund Policy and Texas Education Code §54.006. We understand that these types of situations can be a financial hardship for many families. Therefore, we are pleased to make available the Tuition Refund Insurance plan.
Tuition Refund Insurance can help refund your tuition, fees, and room/board charges, up to the policy limits, if you are unable to complete the semester due to a covered medical reason such as an illness, accident, death or mental health issue. This insurance program complements and enhances our school's refund policy and we believe families will benefit from this added protection.
- How do I sign up for the insurance?
- What is the cost of the policy per semester?
- What is covered by Tuition Refund Withdrawal Insurance?
- What if I sign up and then I change my mind and wish not to purchase the insurance?
- If I want to Increase my limits, do I have to buy more coverage prior to the first day of classes?
- Who is eligible for Tuition Refund Insurance Coverage?
- How long does the policy last?
- How do I file a claim?
- Who receives claim payment?
- Additional frequently asked questions?
How Do I sign up for the Insurance?
Students can sign up for the insurance at the time you register for courses. The sign up is included in the Promise to Pay Agreement that you see prior to you registering for each semester on-line via Panthertracks. Look for the sign-up options at the bottom of the promise to pay agreement. You must be signed up at the start of each semester if you wish to be enrolled in the insurance plan.
What is the cost of the policy per semester?
PVAMU and Sallie Mae will offer four (4) policies to choose from depending on the cost of your tuition and fees which is based on residency and being an on campus student. We will offer a $2,500.00 policy per semester at a rate of $49.25(for those enrolled in 1 8 week session only), a $5,000.00 policy per semester at a rate of $98.50, a $10,000.00 policy per semester at a rate of $197.00 per semester, and a $15,000.00 policy per semester at a rate of $295.50 per semester to cover the cost of tuition, fees, meals and housing.
Panther Medical Withdrawal Insurance (Tuition Refund Insurance)
|Policy Coverage Amount (per semester)
|Cost to Student (per semester)
What is covered by Tuition Refund Withdrawal Insurance?
Up to 100% of covered fees up to the chosen policy limits for:
- Withdrawal of the covered student from an academic program due to illness or injury
- Withdrawal of the covered student from an academic program due to emotional, mental or nervous disorder (Two-day hospitalization required).
- Death of the covered student during the dates of coverage
- Death of the tuition payer (if different from the student) that results in the student's withdrawal from an academic program.
Covered fees include tuition & fees, meals, housing, and any balance due to a refund that may have been issued prior to the removal of financial aid due to the withdrawal.
In order to qualify for "medical withdrawal" the student must have gone through the University's official process for medical withdrawal, and must have confirmation from a physician that withdrawing from the academic program is recommended for medical reasons. Withdrawal forms can be obtained from the Registrar's Office or at the following link http://www.pvamu.edu/registrar/forms/
What if I sign up and then I change my mind and wish not to purchase the insurance?
If a student chooses to purchase a policy and decides later that they do not wish to purchase the policy, the student must cancel the policy prior to the 1st class day of the semester in which the policy was purchased for. Students may submit a request for cancelling the policy in writing by sending an e-mail to email@example.com with ID number and contact phone number or the request may be faxed to 936-261-1959. The request must be received prior to the University's established 1st class day to be processed. If the request is received on or after the 1st class day, the request will be denied.
If I want to Increase my limits, do I have to buy more coverage prior to the first day of classes?
Yes, the student will need to purchase any additional coverage prior to the first day of classes. Requests for additional coverage may be sent to firstname.lastname@example.com. Student must provide their student identification number, contact phone number, and indicate which policy they wish to be moved to in the request.
Who is eligible for Tuition Refund Insurance Coverage?
Any student who is healthy enough to begin classes, attending an accredited academic program that offers this Tuition Refund Insurance program, and responsible for tuition costs, may purchase Tuition Refund Insurance.
How long does the policy last?
The policy begins on the University's first class day and is covered on a semester basis based on the semester the policy is purchased for.
A student can notify Sallie Mae Insurance Services of a claim by calling 800-719-5915 or by emailing email@example.com. Sallie Mae Insurance Services will work with the underwriter, Markel Insurance Company, on behalf of the student to process the claim. The underwriter will issue the payment directly.
Payments will be made directly to the policyholder. Claim payments are made once all of the requested documentation is processed and approved. In most cases, the University is the policyholder and will receive payment on behalf of the insured. If there are no outstanding tuition payments due, the school will forward the payment to the insured. In instances where the University is not the policyholder, payments will be issued to the insured.
Additional frequently asked questions?
Please go to www.gradguard.com/pvamu and click on "frequently asked questions" on the right side of the page.
Was this information helpful? Provide Feedback or report broken links.


text:
49 State Place
Dix Hills, New York, 11743
Commute to Downtown Dix Hills
Most errands require a car.
About your score
49 State Place has a Walk Score of 47 out of 100. This location is a Car-Dependent neighborhood so most errands require a car.
This location is in Dix Hills. Nearby parks include Berkeley Jackson County Park, Manor Plains Nature Park and Caledonia Park.
Explore how far you can travel by car, bus, bike and foot from 49 State Place.
View all Dix Hills apartments on a map.
Search for homes in Dix Hills or find a Dix Hills real estate agent who knows walkable neighborhoods. Links will open on our partner site Redfin.com.
This location is in the city of Dix Hills, NY. Dix Hills has an average Walk Score of 13 and has 26,892 residents.Learn More About Dix Hills

subset: commoncrawl


text:
Budgets and recruiting at EISD
Over the course of the summer months, other EISD administrators and I have written about various school activities and projects that take place during the summer. Each summer has its own unique events, such as this summer's intense preparation related to the initiation of CSCOPE this fall.
Click here to sign up for our E-Edition and read the rest of the story!


text:
PRZOOM - /newswire/ -
Bali, Indonesia, 2010/09/08 - The Export Service Centre - Costs saving eco-friendly service to ensure exported commodities, home décor and fashion goods pest- and rodent-free without using poisonous Methyl bromide gas.
The Export Service Centre (ExportServiceCentre.com), a member of the non-profit Kearny Alliance (kearnyalliance.org), now offers Quarantine Pre-Shipment (QPS®) service at its eco-friendly facility in Denpasar, Bali.
The Export Service Centre-EcO2 (esceco2.com) is a partnership between the Export Service Centre Indonesia and the leading eco-friendly treatment provider, the Netherlands EcO2 B.V. It offers Quarantine Pre-Shipment treatment solutions for commodities, containers and cargo.
Our green QPS facility is critical to the future of Indonesias exports, said Eugene Verspoor, general manager of the Export Service Centre. In 2013, Methyl bromide will be phased out worldwide.
The only alternative without the use of toxic gases is the QPS treatment based on the heat and low oxygen. For exporters, our system is also cost effective, since they do not have to worry about extra costs incurred for clearing containers due to gas levels and they can save demurrage charges.
Weve invested in this proven facility with the worlds leader to ensure Indonesian producers can continue to export and therefore continue to create much-needed jobs in this critical sector.
We look forward to working closely with Balis exporters, fumigators and freight forwarders to offer this service to their customers today and well into the future.
Global environmental concerns bottom line realities for importers
Environmental concerns are top-of-mind in most importing countries. Worries over toxic gas, ozone layer depletion and global warming mean tighter scrutiny of containers and treatment methods.
This means importers may face significant costs if containers are quarantined. Some of these costs include:
Degassing fees of 600, daily demurrage of 45 (plus handling charges);
Delayed delivery which may incur penalties and handling charges;
Customers' complaints over toxic methyl bromide residue;
The ESC-EcO2 service offers, green Quarantine Pre-Shipment service to ensure containers are not quarantined for degassing. More details are available at (esceco2.com);
About The Kearny Alliance
The Kearny Alliance (kearnyalliance.org), a nonprofit 501 (c) (3) foundation based in Scottsdale, Arizona, partners with other international organizations to further its mission of Aid through Trade, to advance international development and poverty alleviation through trade-related business, education, training and applied research.
Key programmatic areas include:
Livelihood development at the grassroots level: Through the social enterprise Export Service Centre (ExportServiceCentre.com) in Indonesia and Cambodia, the Kearny Alliance assists small producers who do not have the capacity to export direct to participate in the global economy. It estimates that in 2009 it created more than 3,800 jobs.
Job creation for small & medium producers: Through the Developing Country Export Assistance Program (kearnyalliance.org/sourcingreports), the Kearny Alliance connects smaller exporters in developing Asia with buyers worldwide. Surveys of beneficiaries show that suppliers have received $440,000 in export orders, and they expect more than $5,000,000 to develop in 2010.
Education & skills development: The Kearny Alliance offers stipend support for students from Myanmar, Cambodia, Vietnam and mainland China to study in Hong Kong and the US. The dozens of Kearny Alliance Scholars receive internships and on-the-job training in export-related companies and organizations in 10 countries.
Trade policy research: One major initiative is FutureofUSChinaTrade.Com, an online center for data, analysis, and insightful discussion on U.S. - China trade offering Forum Discussions News & Analysis, Facts & Figures, Discussion Outcomes, Book reviews and Meet the Experts. This developed from the The Kearny Alliance Arizona State University forums on China, trade and the world economic order high-level series of interlinked meetings held in Phoenix and Beijing which explored potential solutions to issues and challenges of mutual benefit to China-US trade relations.
Agency / Source: The Kearny Alliance
Availability: All Regions (Including Int'l)
Traffic Booster: [/] Quick PRZOOM - Press & Newswire Visibility Checker
Distribution / Indexing: [+]


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Jason Brian Carter (born September 23, 1960) is an English actor, best known for his role as Ranger Marcus Cole on the science fiction television series Babylon 5.
Born in London and raised in Gainsborough, a small market-town in Lincolnshire, Carter appeared on stage since he was a child. Advised to pursue a career in rubber technology, Carter opted instead for three years at London Academy of Music and Dramatic Art. In 1982, he landed his first television role on BBC2's long-running Jackanory (1965–1996) as Hawkwing. He has appeared in numerous television series including Viper, Beverly Hills, 90210, 3rd Rock from the Sun, Angel and Babylon 5. He also played the hedgehog, Hans, in an episode of the television series The Storyteller. Carter also loaned his voice briefly to the James Bond video game GoldenEye: Rogue Agent, in the role of Bond.


text: On Dec 13, 2009, at 1:31 AM, Pierre GM wrote: > On Dec 13, 2009, at 12:11 AM, Robert Ferrell wrote: >> Have you considered creating a TimeSeries for each data series, and >> then putting them all together in a dict, keyed by symbol? > > That's an idea > >> One disadvantage of one big monster numpy array for all the series is >> that not all series may have a full set of 1800 data points. So the >> array isn't really nicely rectangular. > > Bah, there's adjust_endpoints to take care of that. Maybe this will work for the OP. In my work, if a series is missing data the desirable thing is to use the data I have. I don't' want to truncate existing series to fit the short ones, nor pad to fit the long ones. Really depends on the analysis the OP is trying to do.


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Not able to see the usb device instance in minicom for uclinux target
I am new to this post,Currently I am working on a target for which I need to connect a usb keyboard and should be able to input on the console of the device.The details are:
Target board:uclinux for coldfire processor.
when I connect through the serial port and see the filesystem through minicom,I couldnot see the usbdevice instance in /dev of uclinux but there are messages of connection and disconnection(drivers are available in uclinux for the keyboard).I have to input through the keyboard.Can anyone plz help me in approaching the issue.
I will rephrase your question to make sure I understand it correctly.
You have a ucLinux based board with a USB-attached keyboard. The ucLinux based board also has a serial port which you can use to login from a serial terminal of some sort (minicom running on some other host). You want to be able to use the USB keyboard as the standard input component of your serial login?
In order to do this, I think you will have to craft your own getty process that switches its standard input upon completion of the login process. You would probably want to start by writing some code to open and read the keyboard as a device, reading from /dev/whatever_the_USB_keyboard_is. Once you understand how the keyboard sends its data, you can build this into your_getty.
--- rod.
Yes you are right.But "You want to be able to use the USB keyboard as the standard input component of your serial login?" this is not right.I have to input through the keyboard to the device it is attached.Basically it has two processors and one is running uclinux kernel and the other is running the bootloader and UI of the device so that we can input our data into it.Here I can see the dmesg messages showing drivers are installed.
Current status: I can input the characters through the keyboard seen on the minicom terminal.Now I have to capture the characters going to terminal and place them on the GUI specific fields on the device.That is I have to communicate from one processor to another processor.
Okay, your description seems to have some holes. A processor running a bootloader? To boot what?
Are you saying that you have a whole separate CPU that is reading the USB keyboard and writing to the 'GUI'? If I understand this correctly, you essentially have a CPU of some sort that is performing the function of a terminal. It uses a USB keyboard and some kind of display device. What physical connection is there between this second device and your coldfire ucLinux host? Is this second CPU also running Linux?
--- rod.
|All times are GMT -5. The time now is 11:38 AM.


text:
Division of Wildlife dodges a couple of legislative bullets.
The legislative session isn't quite half over and already the Division of Wildlife has dodged a couple of errant bullets.
House committees shot down two proposed bill, one from Rep. Wes McKinley, D-Walsh, and the other from Sen. Greg Brophy, R-Wray, that would have prohibited or delayed the Colorado Oil and Gas Conservation Commission from adhering to House Bill 1298, the Colorado Wildlife Habitat Stewardship Act, which passed the 2007 Legislature by a unanimous vote.
Still to be decided is the fate of House Bill 1255, another HB 1298-related bill that's more of an issue with the Department of Natural Resources since it includes the Colorado Oil and Gas Conservation Commission.
This one is sponsored by Rep. Cory Gardner, R-Yuma. It would strip the Division of Wildlife of its consulting role in the effects of energy development on wildlife and wildlife habitat.
"This bill would diminish the ability of the Division of Wildlife to recommend protections that were clearly articulated in House Bill 1298," said DNR spokesman Theo Stein. "It would also undermine 18 months of dialogue and compromise that took place during the rule-making process."
The bill also says a landowner cannot be denied a drilling permit if he or she refuses to undergo a wildlife consultation aimed at what 1298 called "best management practices" in terms of wildlife habitat.
This bill was being debated Tuesday in the House Agriculture and Natural Resources Committee.
The other major bill that's appeared this session is Senate Bill 24, carried by Sen. Al White, R-Steamboat Springs. In its original form this bill would have changed drastically the state's game damage program and, some sportsmen argued, would have contributed to making hunting a game for the rich by allowing landowners to charge trespass fees of up to $2,500 per gun and still get game-damage payments.
The DOW has been working hard on this bill to retain the $100 fee limit and eliminate a demand from landowners for mandatory kill permits when they thought the division wasn't responsive enough to game damage problems.
According to Division spokesman Tyler Baskfield, the agency agreed to make elk-resistant panels and other prevention materials available to more landowners and to provide permanent damage-prevention materials to landowners.
Baskfield said current amendments allow landowners charging more than $100 limit to received game-damage payments only after going through their local Habitat Partnership Program committee.
"If the HPP can't find a solution, the landowners can turn to us," Baskfield said.
Sportsmen still aren't happy about this last provision, since that means landowners who charge high prices for public access still can get damage materials funded by license fees.
This bill is set for a hearing by the Senate appropriations committee in the next two weeks.
Other legislation of note:
House Bill 1071: Initially sought to raise search and rescue fee surcharges to $1 on every license and habitat stamp sold and allow municipal police departments to apply for search and rescue funds. As of now, the fee increase has been cut to 50 cents and municipal police are not eligible for search and rescue funds.
Pro: More money for reimbursing equipment and training costs to under-funded sheriff's departments. Con: Hunters and anglers still pay nearly two-thirds of the cost, yet only need 9 percent of the rescues.
Still undecided is how to get unlicensed outdoors users to pay their share.
The sponsor, Rep. Sara Gagliardi, D-Arvada, wants the DOW to be more pro-active in promoting the Colorado Search and Rescue Card.
Colorado Habitat Stamp: This legislation isn't due for renewal until 2010, but it takes time for something like this, with all the politics that come into play, to make its way through the Legislature.
Several big changes are pending. Foremost is how and where the money is spent. Again, hunters and anglers supply more than 98 percent of the funding, another inequitable breakdown.
Conservation groups initially pledged their support for the legislation, but some of those groups, and this includes many anglers, aren't happy about the wording that requires 60 percent of the money to be spent on big-game habitat.
One possible compromise is to delete that limiting language and allow the DOW more leeway in deciding how to spend the money. That is, buy property when it's available, without being tied to a pre-decided purpose.
"We've been meeting with environmental groups and sportsmen groups trying to come up with a way to package the habitat stamp," Baskfield said. "We've been making some progress on this one."
Other possible changes: Eliminate the two $5 stamps in favor of one $10 stamp; require anglers to buy the $10 stamp on their third and subsequent one-day fishing licenses; and remove the requirement that anyone using a State Wildlife Area must have a habitat stamp.


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The transactivation of nuclear receptors is regulated by both ligand binding and phosphorylation. We previously showed that RARα (retinoic acid receptor α) phosphorylation by c-Jun N-terminal kinase contributes to retinoid resistance in a subset of NSCLC cells (non-small cell lung cancer cells), but the aetiology of this resistance in the remainder has not been fully elucidated [Srinivas, Juroske, Kalyankrishna, Cody, Price, Xu, Narayanan, Weigel and Kurie (2005) Mol. Cell. Biol. 25, 1054–1069]. In the present study, we report that Akt, which is constitutively activated in NSCLC cells, phosphorylates RARα and inhibits its transactivation. Biochemical and functional analyses showed that Akt interacts with RARα and phosphorylates the Ser96 residue of its DNA-binding domain. Mutation of Ser96 to alanine abrogated the suppressive effect of Akt. Overexpression of a dominant-negative form of Akt in an NSCLC cell line decreased RAR phosphorylation, increased RAR transactivation and enhanced the growth-inhibitory effects of an RAR ligand. The findings presented here show that Akt inhibits RAR transactivation and contributes to retinoid resistance in a subset of NSCLC cells.
- Akt
- non-small cell lung cancer cell (NSCLC cell)
- phosphorylation
- retinoic acid receptor α (RARα)
- retinoid resistance
- transactivation
RARs [RA (retinoic acid) receptors] are ligand-dependent transcription factors that regulate genes involved in various biological processes, such as pattern formation, reproduction, cell differentiation and maintenance of tissue homoeostasis [1,2]. RARs and RXRs (retinoid X receptors) bind to their target response elements as heterodimers and regulate gene expression in response to their ligands, all-trans RA and 9-cis RA [3]. In the absence of ligand, RXR–RAR heterodimers associate with a multiprotein complex containing transcriptional co-repressors that induce histone deacetylation, chromatin condensation and transcriptional suppression. Ligand binding causes the receptors to dissociate from the co-repressors and associate with co-activators that have histone acetyltransferase activity and induce local chromatin decondensation, recruitment of the RNA polymerase II holoenzyme and activation of target gene transcription [4,5].
Three major classes of co-activators have been studied in detail regarding their role in co-activating nuclear receptors. Members of the Swi/Snf/Brg class are involved in ATP-dependent remodelling of chromatin [6]. Members of the second class, which includes CBP [CREB (cAMP-response-element-binding protein)-binding protein]/p300, P/CAF (p300/CBP-associated factor) and p160/SRC (steroid receptor co-activator) have intrinsic histone acetyltransferase activity and have a crucial role in histone modification of transcriptionally active genes [7]. Members of the third class belong to TRAP (thyroid hormone receptor-associated protein)/DRIP (vitamin D receptor-interacting protein)/ARC (activator-recruited cofactor) complexes, interact with ligand-bound nuclear receptors and stimulate transcription of in vitro DNA templates [8]. Co-activators contain one to several copies of leucine-rich LXXLL motifs in their receptor interaction domains. Crystallographic and biochemical studies have revealed that these LXXLL motifs interact with the ligand-activated AF-2 (activation function-2) domain of nuclear receptors, thereby providing a molecular basis for nuclear receptor-co-activator recruitment [9–11].
In addition to being activated by RA, RAR transactivation is regulated by phosphorylation. RARs are substrates for a variety of serine/threonine kinases, including PKA (protein kinase A), PKC, CDK7 (cyclin-dependent kinase 7) and p38 [12–16]. PKC phosphorylates RARα and strongly reduces its transactivation by inhibiting its dimerization with RXRα [13], whereas CDK7 increases RAR transactivation by phosphorylating its A/B domain [14]. We have shown that activation of JNK (c-Jun N-terminal kinase) by stress signals leads to phosphorylation of RARα, resulting in ubiquitin-mediated proteasomal degradation of the receptor [17]. Another serine/threonine kinase previously shown to phosphorylate nuclear receptors is PKB/Akt, a downstream mediator of class I PI3K (phosphoinositide 3-kinase) [18,19]. Akt phosphorylates the AF-1 domain of oestrogen receptor-α, thereby increasing its activity in a ligand-independent manner [20]. In contrast, Akt suppresses the transactivation of the androgen receptor [21]. Beyond its role in regulating nuclear receptor function, Akt is the product of a proto-oncogene and phosphorylates a number of substrates that are important regulators of cell survival and transformation [22].
We previously showed that RARα is aberrantly phosphorylated in lung cancer cells by JNK and that RARα phosphorylation contributes to the blockade of retinoid signalling observed in a subset of lung cancers [17]. However, JNK is not responsible for the retinoid signalling defects observed in most of the retinoid-resistant NSCLC (non-small cell lung cancer) cell lines. Retinoids have demonstrated efficacy in the treatment of certain cancers but are ineffective in the treatment of lung cancer patients in part due to a defect in the transactivation properties of RARs in lung cancer cells, the aetiology of which has not been fully defined [23]. In the present study, we examined whether this retinoid resistance is instead mediated by Akt, which is constitutively active in NSCLC cell lines through amplification of genes encoding the p110α catalytic subunit of PI3K (PI3KCA) and ErbB family members, epigenetic silencing of the PTEN gene and activating mutations in the genes PI3KCA, KRAS and EGFR [24–27]. We found that Akt phosphorylates RARα at Ser96 and inhibits its transactivation. We also found that inhibition of Akt enhances RAR activity and the growth-inhibitory effects of retinoids on NSCLC cells. Together, our findings support the hypothesis that Akt contributes to the retinoid resistance commonly found in NSCLC cells.
We purchased all-trans RA, TTNPB {E-4-[2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthalenyl)-1-propenyl]benzoic acid}, 9-cis RA and IGF-I (insulin-like growth factor-I) from Sigma–Aldrich (St. Louis, MO, U.S.A.); [γ-32P]ATP, [32P]Pi and [32P]dCTP from ICN Biomedicals (Costa Mesa, CA, U.S.A.); antibodies against human RARα and PARP [poly(ADP-ribose) polymerase] from Santa Cruz Biotechnology (Santa Cruz, CA, U.S.A.); polyclonal antibodies against Akt, Ser473-phosphorylated Akt (P-Akt), Ser21/Ser9-phosphorylated GSK3 (glycogen synthase kinase 3), GSK3, Thr389-phosphorylated p70S6K (p70 S6 kinase), p70S6K and PTEN (phosphatase and tensin homologue deleted from chromosome 10) from Cell Signaling Technology (Beverly, MA, U.S.A.); monoclonal antibodies against FLAG and tubulin from Sigma–Aldrich; anti-HA (haemagglutinin) antibody from Roche Molecular Biochemicals (Indianapolis, IN, U.S.A.); and purified recombinant phosphorylated and unphosphorylated Akt from Upstate Biotechnology (Lake Placid, NY, U.S.A.).
Cell culture
COS-1, HEK-293T (human embryonic kidney 293T), HeLa and NSCLC cells (H1299, Calu-1 and H460) were purchased from the American Type Culture Collection (Manassas, VA, U.S.A.). COS-1, HEK-293T and HeLa cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% (v/v) fetal bovine serum at 37 °C. NSCLC cells were maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum at 37 °C.
cDNA constructs
HA–Akt, HA–Akt (T308D/S473D or DD mutant) and HA–Akt (K179A/T308A/S473A or AAA mutant) expression vectors were gifts from Dr Gordon Mills (M. D. Anderson Cancer Center). The Myr–Akt (where Myr stands for myristoylation) vector was constructed by inserting the myristoylation/palmitoylation signal of Lck tyrosine kinase [28] into the N-terminus of Akt. The FLAG–RARα, RARβ-luc, DR5TK-luc (where DR5 is direct repeat separated by 5 nt, and TK is thymidine kinase), TK-luc, GALTK-luc (where GAL is GAL4 DNA-binding sequence) and GST (glutathione S-transferase)–RARα deletion constructs have been described elsewhere [17]. The GAL–RARα (where GAL is yeast GAL4 protein) expression plasmids were constructed by cloning the cDNA encoding RARα into the pFA-CMV vector (Stratagene, La Jolla, CA, U.S.A.). The S95A, S96A, S115A, S154A and S157A mutations were introduced into the GAL–RARα by site-directed mutagenesis using the QuikChange® mutagenesis kit (Stratagene).
Purification of GST-tagged proteins
GST-tagged wild-type RARα and deletion mutants were expressed and purified from the BL21 bacterial strain (Stratagene) according to manufacturer's instructions. The integrity of the purified proteins was assayed by Coomassie Blue staining and Western blotting with anti-GST antibodies.
In vivo labelling and in vitro kinase assay
For the in vivo labelling experiments in HeLa cells, Lipofectamine™ 2000 (Invitrogen Life Technologies, Carlsbad, CA, U.S.A.) was used to co-transfect HeLa cells (1.5×106 cells per 10 cm dish) with plasmids expressing HA–Akt or Myr–Akt and FLAG-tagged, wild-type RARα. After 24 h, the cells were serum-starved overnight in phosphate-free medium and then incubated in the same medium for 4 h in the presence of 250 μCi/ml [32P]Pi. Where indicated in the Figure legends, the cells were exposed to IGF-I (100 ng/ml) 1 h before harvesting. Cells were lysed in radio-immunoprecipitation assay buffer containing 100 nM okadaic acid, 1 mM orthovanadate and protease inhibitor cocktail (Sigma–Aldrich). FLAG-tagged RARα was immunoprecipitated with anti-FLAG antibodies and the immunoprecipitates were subjected to electrophoresis and autoradiography. For in vivo labelling experiments in Calu-1 cells, cells were incubated with 250 μCi/ml [32P]Pi for 8 h and RARα was immunoprecipitated with anti-RARα antibodies.
For the in vitro kinase assays, recombinant active Akt was incubated with 2 μg of substrate proteins in kinase buffer containing 25 mM Hepes, 25 mM MgCl2, 25 mM β-glycerophosphate, 20 μM ATP, 0.5 mM dithiothreitol and 10 μCi of [γ-32P]ATP for 30 min at 37 °C. The labelled proteins were separated by SDS/PAGE and detected by autoradiography.
For stoichiometric analysis of phosphorylation, 0.75 μg of purified GST–RARα was incubated with increasing amounts of recombinant active Akt in kinase buffer containing 5 μCi of [γ-32P]ATP for 30 min. The labelled proteins were separated by SDS/PAGE. A GST–RARα band that exhibited saturated levels of phosphorylation was excised from the gel, and radioactivity in the protein was quantified by liquid-scintillation counting. The amount (mol) of phosphate incorporated per mol of GST–RARα was calculated.
Phospho amino acid analysis
GST–RARα DBD (DNA-binding domain) (5 μg) was subjected to in vitro Akt kinase reaction and the labelled protein was separated by SDS/PAGE. The phosphorylated band corresponding to GST–RARα DBD, visualized by autoradiography, was excised and phospho amino acid analysis was performed as described previously [29].
Luciferase assays
COS-1, HEK-293T and NSCLC cells (8×104 cells/well) were transfected in 24-well plates with the DR5TK-luc (200 ng), RARβ-luc (200 ng) or GALTK-luc (50 ng) reporter constructs, along with plasmids expressing Akt or GAL–RAR fusion proteins (50 ng). pRLTK-luc (50 ng) was transfected as an internal control. The total amount of plasmid was kept to 1 μg/well. Cells were serum-starved overnight 24 h later and treated with all-trans RA or TTNPB (10−6 M) for 15 h, and cell lysates were assayed for luciferase activity using a dual-luciferase reporter assay system (Promega, Madison, WI, U.S.A.). Results were expressed as relative luciferase activity, the ratio of firefly to Renilla luciferase activity. Each experiment was performed three times, and results from a representative experiment are shown as the means±S.D. (indicated by error bars) from three identical wells.
EMSA (electrophoretic mobility-shift assay)
DR5 oligonucleotides (5′-AGCTTAAGGGTTCACCGAAAG-TTCACTCGCAT-3′) and an NF-κB (nuclear factor κB) binding site used as non-specific oligos (5′-TTGGGGGAAGGG-GGAATCTCTAGGCAAAGG-3′) were used in the present study. Recombinant GST–RARα (40 ng) and in vitro-translated RXRα were incubated with 32P-labelled DR5 oligos for 30 min at 4 °C in a binding buffer (10 mM Tris/HCl, 40 mM KCl, 6%, v/v, glycerol, 1 mM dithiothreitol and 0.05% Nonidet P40). Where indicated in the Figure legends, GST–RARα was pre-incubated with active Akt kinase for 20 min in the presence of 1 mM ATP at 30 °C. The RXR–RAR heterodimers bound to an RARE (RA response element) were resolved on 6% (w/v) polyacrylamide gels and subjected to autoradiography.
GST pull-down assays
GST and wild-type GST–RARα (3 μg) were immobilized on glutathione–agarose beads and incubated with purified Akt for 30 min on ice in binding buffer [buffer A (25 mM Tris/HCl, 150 mM NaCl and 0.1% Triton X-100)]. Protein complexes were washed twice with the same buffer, and bound materials were resolved by SDS/PAGE and subjected to Western blotting with anti-Akt antibodies.
Co-immunoprecipitation experiments
To study Akt–RAR interactions, COS-1 cells were transfected with plasmids expressing HA–Akt and FLAG–RARα. After 48 h, the cells were lysed in buffer A, and the lysates were incubated with anti-FLAG or control IgG antibodies at 4 °C. The immunoprecipitates were washed twice with buffer A and subjected to Western blotting with anti-HA antibodies.
Generation of stably transfected cell lines
H1299 and Calu-1 NSCLC cells were transfected with HA–Akt(DD) and HA–Akt(AAA) constructs respectively using Lipofectamine™ (Invitrogen). Transfectants were selected in G418 (600 μg/ml) for 2 weeks, and individual clones were isolated and later analysed for expression of transfected gene products.
Quantitative PCR
H1299 cells stably transfected with empty vector or Akt(DD) were treated with RA (10−6 M) for 5 h. The cells were harvested, and total RNA was extracted using an RNA isolation kit (Qiagen, Valencia, CA, U.S.A.). Reverse-transcriptase reactions were performed with 1 μg of total cellular RNA using specific reverse primers for RARβ and cyclophilin. The reverse-transcriptase reaction (10 μl) was added to 40 μl of PCR mixture containing 1×PCR buffer (20 mM Tris, pH 8.4, and 50 mM KCl), 400 nM primers, 5 mM MgCl2, ROX reference dye (Invitrogen), Taq polymerase and 100 nM fluorogenic probe labelled with 6-carboxyfluorescein and a quencher. Amplification and quantification were based on real-time monitoring of probe degradation and were carried out using an ABI Prism 7700 performing 40 cycles at 94 °C for 12 s and 60 °C for 30 s. RARβ transcript levels were normalized to those of cyclophilin mRNA, which was used as a control.
Cell proliferation assays
Calu-1 cells (1×103) stably transfected with empty vector or Akt(AAA) construct were grown in RPMI 1640 medium containing 1% fetal bovine serum in 96-well plates. After 24 h, the cells were treated for 8 days with different concentrations of TTNPB as indicated in the Figure. Ligand was replenished every 48 h. Relative cell density was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay, as described elsewhere [30].
Statistical analysis
Statistical analyses were performed using one-way ANOVA test for luciferase assays and two-way ANOVA test for cell proliferation assays.
Akt represses ligand-dependent activation of retinoid receptors
We began investigating the effect of Akt on retinoid signalling with HEK-293T cells, because they express functional RAR and RXR proteins that transcriptionally activate RAREs in response to ligand [31]. We transiently transfected HEK-293T cells with a reporter construct containing the DR5 response element to which the RXR–RAR heterodimeric complex binds. Treatment with the RAR ligand all-trans RA led to a 5-fold increase in DR5TK-luc activity, whereas the activity of the control TK-luc reporter was unchanged (Figure 1A). Co-transfection of increasing amounts of Myr–Akt, an activated form of Akt [28], inhibited RA-induced DR5TK-luc activity in a dose-dependent manner (Figure 1A). To control for the possibility that PH domains suppressed RAR transactivation, we transfected HEK-293T cells with dominant-negative Akt (AAA mutant), which had no effect on DR5TK activity (Figure 1A).
We also co-transfected HEK-293T cells with Akt and a reporter construct driven by the RARB gene promoter, which contains the DR5 response element. Myr–Akt, but not Akt(AAA), suppressed RA-induced activation in those cells, suggesting that Akt represses the DR5 response element in the context of a natural promoter (Figure 1B). Stable transfection of H1299 NSCLC cells with Akt(DD), another constitutively active form of Akt, stimulated Akt-dependent signalling, as shown by increased expression of P-GSK3 (phosphorylated GSK3) (Figure 1C), and suppressed RA-induced RARB expression (Figure 1D). Together, these findings indicate that activation of Akt inhibits ligand-dependent activation of retinoid receptors.
Akt phosphorylates DBD of RARα
We performed in vivo labelling experiments with [32P]Pi to determine whether Akt activation increases RARα phosphorylation in cells. RARα phosphorylation increased in cells transfected with Myr–Akt (Figure 2A). To determine whether RARα is a substrate of Akt, we looked for evidence of physical interactions between the two. In COS-1 cells, HA-tagged Akt associated with FLAG-tagged RARα (Figure 2B). In vitro kinase assays showed that purified, active Akt phosphorylates GST–RARα (Figure 2C) and purified Akt kinase interacted with GST–RARα in vitro (Figure 2D). Stoichiometric analysis of the in vitro kinase reaction revealed that 33% of the purified RARα protein was phosphorylated by Akt.
We next used deletion mutants to determine the regions of RARα that are phosphorylated by Akt. Akt phosphorylated the N-terminus (residues 1–187) and DBD, but not the LBD (ligand-binding domain), of RARα (Figure 2C). Phospho amino acid analysis of RARα DBD revealed phosphoserine residues, consistent with Ser/Thr-specific kinase activity of Akt (Figure 2E).
DBD of RARα is required for Akt-mediated repression
The inhibition of nuclear receptor function by phosphorylation may involve mechanisms that reduce DNA-binding activity, nuclear export, or stability of the receptor [32–34]. We investigated the effect of Akt on the DNA-binding activity of RARs by performing EMSAs. Recombinant RARα was subjected to in vitro kinase reactions with Akt and then incubated with in vitro-translated RXRα and 32P-labelled DR5 probe. We did not observe any change in the DNA-binding activity of the RXR–RAR heterodimer after the kinase reaction (Figure 3A).
To examine the effect of Akt activation on RARα protein stability and localization, we transiently transfected HEK-293T cells with Myr–Akt. Nuclear and cytosolic fractions of the cells were prepared, and RARα protein levels were assayed. Myr–Akt expression did not alter the protein levels in the nuclear fraction or increase the cytoplasmic localization of RARα (Figure 3B). PARP and α-tubulin were used as loading controls for nuclear and cytosolic fractions respectively. Taken together, these results suggest that the down-regulation of RARα function by Akt does not involve changes in DNA-binding activity, stability, or localization of RARα.
We next examined whether Akt would repress RARα transactivation when fused to a heterologous DBD. We transfected HEK-293T cells with a GAL4 DBD–RARα expression plasmid, along with a reporter construct containing a GAL4 response element (GALTK-luc). Because RXR is not required, we used TTNPB, a synthetic RAR ligand, to transactivate GAL–RAR. Expression of Myr–Akt substantially inhibited ligand-induced transactivation of GAL–RARα but not GAL4 (Figure 3D). Myr–Akt had no effect on GAL–RXRα function (Figure 3E), suggesting that Akt represses retinoid signalling through RAR.
RARα contains two activation domains (AF-1 and AF-2) that are required for maximal activation by its ligand. AF-1 and AF-2 are within the A/B region and LBD of RARα respectively. We investigated which of these domains is required for the suppression of RARα transactivation by Akt. We transiently co-transfected HEK-293T cells with Myr–Akt along with GAL–RAR ΔAB (which lacks the AF-1 domain) or GAL–RAR ΔAB/DBD (which lacks both DBD and AF-1 domain). In the absence of Myr–Akt, compared with wild-type RARα, the transactivation of GAL–RAR ΔAB was decreased in the presence of ligand. This was expected because of the known synergy between the AF-1 and AF-2 domains [35,36]. The transactivation of GAL–RAR ΔAB was repressed by Myr–Akt, whereas the transactivation of the ΔAB/DBD RAR construct was resistant to down-regulation by Akt (Figure 3D), suggesting that the DBD is required for repression by Akt. To confirm this further, we transfected HEK-293T cells with GAL–RAR ΔDBD, a construct that lacks the DBD but retains both AF-1 and AF-2 domains. Myr–Akt failed to repress the transactivation of GAL–RAR ΔDBD (Figure 3D). To control for the possibility that PH domains suppressed GAL4 chimaeric protein transactivation, we transfected HEK-293T cells with Akt(AAA), which had no effect on wild-type GAL–RARα or GAL–RAR ΔDBD (Figure 3D).
Ser96 is an Akt phosphorylation site
Sequence analysis of the DBD revealed no consensus Akt phosphorylation sites. We therefore created a series of GAL–RARα constructs in which all the serine residues in this region were individually mutated to alanine residues, and we analysed the constructs for transactivation function. Myr–Akt suppressed ligand-induced transactivation of all GAL–RARα mutant constructs except the S96A mutant (Figure 4A). In the in vitro kinase assays using RARα DBD as the substrate, Akt phosphorylated wild-type GST–DBD but not the S96A mutant (Figure 4C), indicating that Ser96 is an Akt phosphorylation site.
To investigate the functional significance of Ser96 phosphorylation in the context of RXR–RAR heterodimers, we examined the ability of the S96A mutation to block Akt-mediated repression of DR5 transactivation by RXR–RAR heterodimers. We observed that, in the context of a DR5 element, ligand-induced transactivation of RARα S96A mutant was less than that of wild-type RARα (Figure 4D), which was not the case in the setting of GAL chimaeric constructs (Figure 4A). Furthermore, Myr–Akt repressed ligand-induced DR5 transactivation in COS-1 cells co-transfected with wild-type RARα but not those co-transfected with RARα S96A (Figure 4D). EMSA revealed that RARα S96A bound to a DR5 element and heterodimerized with RXRα as well as wild-type RARα did (Figure 4E), indicating that the reduced DR5 transactivation by RARα S96A was not due to impaired DNA-binding or heterodimerization properties. Sequence analysis of RARβ and RARγ revealed that Ser96 is a conserved residue (Figure 4F). We therefore postulated that Akt also inhibits these RAR subtypes. Indeed, Myr–Akt, but not Akt(AAA), repressed transactivation of GAL–RARβ in HEK-293T cells (Figure 4G). Together, these findings indicate that RAR Ser96 phosphorylation is required for the suppression of RXR–RAR transactivation by Akt.
Akt contributes to retinoid resistance in NSCLC cells
Although many NSCLC cells express normal levels of retinoid receptors, they are not sensitive to the biological effects of RA in part because of a defect in RAR transactivation function [23]. To test our hypothesis that high Akt activity contributes to RAR dysfunction in NSCLC cells, we inhibited Akt signalling in two NSCLC cell lines, Calu-1 and H460, which differed with respect to basal Akt activation, as shown by Western blotting of P-Akt (Figure 5A). Akt is constitutively phosphorylated in Calu-1 cells in part due to its low expression of the PTEN lipid phosphatase [37]. Inhibition of Akt by transient transfection of the dominant-negative mutant Akt(AAA) increased DR5TK activity in Calu-1 cells but not in H460 cells (Figure 5B), supporting a role for Akt in the suppression of RAR function in Calu-1 cells.
Finally, we examined whether the high Akt activity in Calu-1 cells contributes to the resistance of those cells to growth inhibition by the RAR ligand. Calu-1 cells stably transfected with Akt(AAA) displayed a block in Akt signalling, as shown by Western blotting for P-p70S6K (Figure 5C). Metabolic labelling studies revealed that RARα was phosphorylated to a lesser extent in Calu-1 Akt(AAA) transfectants than controls (empty vector transfectants) (Figure 5D). Compared with its effect on controls, TTNPB more profoundly inhibited the proliferation of Akt(AAA) transfectants (Figure 5E). Together, these findings support the hypothesis that Akt contributes to retinoid resistance in a subset of NSCLC cells.
In the present study, we explored the role of Akt in the regulation of RAR function and found that Akt phosphorylates RARα and suppresses its transactivation. We identified an Akt phosphorylation site on Ser96 within the DBD of RARα. Furthermore, Akt inhibition enhanced retinoid sensitivity in an NSCLC cell line. These findings indicate that RAR is a novel Akt substrate, that Akt is important in the regulation of RAR function, and that constitutive Akt activation may contribute to retinoid resistance in a subset of NSCLC cells.
Several lines of evidence suggest that RARα is a direct substrate of Akt. We showed that RARα physically associates with Akt in vitro (Figure 2D) and in vivo (Figure 2B). Akt also increased the phosphorylation of RARα in vitro (Figure 2C) and in vivo (Figure 2A). Mutation of Ser96 of the DBD to an alanine residue prevented Akt-mediated phosphorylation (Figure 4C) and restored transactivation (Figure 4A). Finally, Ser96 is conserved across species and RAR subtypes (Figure 4F). These results suggest that Ser96 is a bona fide Akt phosphorylation site. However, Ser96 does not resemble the consensus Akt phosphorylation site (RXRXXS/T) [38], and we do not know the mechanism by which Akt recognizes this region. There are other examples of Akt substrates that do not have the consensus motif, such as CREB and YB-1 (Y-box binding protein-1) [39,40]. Furthermore, although mitogen-activated protein kinases usually phosphorylate serine or threonine residues that are followed by proline, there are several physiological substrates such as TAB1 [TAK-1 (transforming growth factor-β-activated kinase-1) binding protein], cdc25 and microtubule-associated protein Tau where this is not observed [41–43].
We showed that the DBD of RARα is important for mediating repression by Akt (Figure 3D), indicating that the DBD has functions other than DNA binding. Although nuclear receptors usually interact with co-regulatory proteins through their LBDs, there are several reports of DBDs being used as docking sites for co-activator recruitment. For example, Jun dimerization protein-2 interacts with the DBD of the progesterone receptor and stimulates AF-1 function by recruiting other co-activators such as CBP and P/CAF [44]. Similarly, the PPARγ (peroxisome-proliferator-activated receptor γ) co-activator PGC-1 (PPARγ co-activator-1) interacts with the DBD of PPARγ. This interaction induces a conformational change in PGC-1 that enables its recruitment of other co-activators, such as SRC-1 and CBP/p300 [45]. Along similar lines, P/CAF has also been shown to interact with the DBDs of RXR and RAR in vitro. This RAR–P/CAF interaction potentiates the ligand-dependent transactivation of RA-responsive reporter constructs [46,47]. Thus we have identified a novel role of the DBD in RAR transactivation. While the mechanism has not been fully defined, one possibility is that RARα phosphorylation by Akt inhibits the ability of RAR to recruit essential co-activators to the receptor complex.
Whereas normal human bronchial epithelial cells are sensitive to the growth-inhibitory effects of retinoids, NSCLC cells are typically resistant. Multiple mechanisms contribute to retinoid resistance in NSCLC cells. First, expression of RARB, which has tumour-suppressing properties [48,49], is silenced in certain NSCLC cell lines through DNA methylation and aberrant histone acetylation on the gene's promoter [50,51]. Secondly, we previously showed that RAR phosphorylation by JNK decreases RAR levels by inhibiting RAR stability in H322 NSCLC cells [17]. Thirdly, certain NSCLC cell lines are resistant to retinoids despite having high levels of retinoid receptors, but the receptors in those cells have transactivation defects [23]. Fourthly, we previously showed that certain retinoid-resistant NSCLC cells, including the Calu-1 cells studied in the present study, express co-activators at levels similar to those in retinoid-sensitive cells, suggesting that aberrant expression of co-activators does not have a role in retinoid resistance [51]. In the present study, we show that co-activator function may be aberrant in NSCLC cells with high Akt activity, thereby contributing to retinoid resistance.
This work was supported in part by National Institutes of Health grant number R01CA80686.
Abbreviations: AF, activation function; CREB, cAMP-response-element-binding protein; CBP, CREB-binding protein; CDK7, cyclin-dependent kinase 7; DBD, DNA-binding domain; DR5, direct repeat separated by 5 nt; EMSA, electrophoretic mobility-shift assay; GSK3, glycogen synthase kinase 3; GST, glutathione S-transferase; HA, haemagglutinin; HEK-293T, cells, human embryonic kidney 293T cells; IGF-I, insulin-like growth factor-I; JNK, c-Jun N-terminal kinase; LBD, ligand-binding domain; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; NSCLC, cell, non-small cell lung cancer cell; p70S6K, p70 S6 kinase; PARP, poly(ADP-ribose) polymerase; P/CAF, p300/CBP-associated factor; P-GSK3, phosphorylated GSK3; PPARγ, peroxisome-proliferator-activated receptor γ; PGC-1, PPARγ co-activator-1; PI3K, phosphoinositide 3-kinase; PKA, protein kinase A; PTEN, phosphatase and tensin homologue deleted from chromosome 10; RA, retinoic acid; RAR, RA receptor; RARE, RA response element; RXR, retinoid X receptor; SRC, steroid receptor co-activator; TK, thymidine kinase; TTNPB, E-4-[2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthalenyl)-1-propenyl]benzoic acid
- The Biochemical Society, London


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We have already bid a fond adieu to summer, here in Copenhagen and are getting used to the cloudy skies and the crisp air that are common trademarks of the Danish autumn. But I couldn't resist making one last summery treat before getting around to the winter warmers.
Pineapple Upside Down cake is one of those retro desserts that you either hate or love, there are no in-betweens. What separates this version from the rest is that it is made with beurre noisette or brown butter. Beurre Noisette is butter that is browned till it lets off a nutty aroma. But this description does nothing to explain the sheer awesomeness that is beurre noisette.
You will be hooked the first time you smell it. If you ever hear it mentioned in a random conversation or on a TV show, you will smile with fond memories of the last time you tasted it. Trust me, it's that good.
This recipe is from Chef John. He even has a video showing the recipe step-by-step. I omitted the cardamom powder and the dark rum, but followed the rest of the recipe. I have made this cake quite a few times and it has been perfect every time. And it comes with the added excitement of turning the baking skillet upside down and waiting to see the perfectly caramelized pineapple slices on top of the cake!
Homemade caramel, sweet, juicy pineapple and a buttery, crumbly, cake. What's not to love?
For good measure, I also made some Pineapple Upside Down cupcakes using the same recipe. The only change to the recipe was that I made the caramel in a saucepan and added it into a muffin pan. The remaining steps are the same.
Make these today and treat yourself to some sunshine!
Here's the recipe:
(Makes 1 12-inch cake or 12 cupcakes)
Prep Time: 15 minutes
Cooking Time: 45 minutes
For the topping:
1/4 cup butter
3/4 cup light brown sugar
1 tablespoon pineapple juice
1/2 small fresh pineapple – peeled, cored and sliced
For the cake:
1 1/2 cups all-purpose flour
1/2 cup butter
1/2 teaspoon salt
2 teaspoons baking powder
1/2 cup white sugar
1 egg
1/2 cup cold milk
Candied Cherries (optional)
Preheat the oven to 200 degrees Celsius.
Melt 1/4 cup butter in a 12-inch cast iron skillet over medium-low heat. Stir in the brown sugar and pineapple juice. Cook over medium-low heat until sugar is dissolved and mixture is bubbling, about 5 minutes.
Remove from heat and spread pineapple slices in a layer over the brown sugar mixture, completely covering the mixture. Set skillet aside.
Melt 1/2 cup butter in small skillet over low heat until the butter begins to brown and release a nutty fragrance, about 5 minutes. Watch carefully, butter burns easily. Remove from heat and let cool for 10 minutes.
Whisk together flour, salt, baking powder, and white sugar in a large bowl. Whisk in egg and cold milk until just combined. Pour in melted butter and stir to mix thoroughly.
Pour batter over the pineapple slices in the skillet; spread evenly to cover.
Bake in preheated oven until the cake begins to bubble around the edges, the top is browned, and a toothpick inserted into the center comes out clean, about 30 to 35 minutes. Let cool in the skillet for 10 minutes.
Loosen the cake from the skillet by running a knife around the inside edge of the cake. Invert a large plate over the top of the skillet and flip over, releasing the cake to show the pineapple slices on top.
Decorate with a few candied cherries, if desired.


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THERE is still limited public access to Romsey War Memorial park following the winter floods.
Test Valley Borough Council chiefs have opened some parts, but others remain out of bounds to the public.
Water levels have gone down, but council officials are still cleaning up after debris and wastewater affected the popular open space, which has Green Flag status.
The children's play area was damaged by floodwater and will need to be repaired before it can be used again.
A council spokesman said: "Romsey War Memorial Park has been partially reopened with some areas cordoned off behind a temporary fence.
"Signs are in place to inform the public that the children's play area is out of action. Further signs remind visitors to skeep to the pathways. The council will continue its efforts to clear up the remaining areas as the water levels drop further."


text: Search Results Studies Variables Publications (1) Your query returned 1 citations. Sort by: Pub Date (newest) Title Recent Additions Author Clear All Pub. Year Citation 1. 2007 Matjasko, Jennifer, Grunden, Leslie, Ernst, Jody L. Structural and dynamic process family risk factors: Consequences for holistic adolescent functioning. Journal of Marriage and Family. 69, (3), 654-674. Full Text Options: DOI Worldcat Google Scholar Export Options: RIS EndNote XML Results 1 - 1


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Symptoms include:
- Fever
- Malaise
- Ongoing, severe back pain that is worse with movement
- Sweating
- Swelling, warmth, redness and tenderness at the site of the infection
- Weight loss
When the infection is in the back, the patient may have back pain or muscle spasms but no fever. If the infection invades the space that surrounds the spinal cord, the patient may have severe, radiating back pain with muscle weakness and paralysis.
If the condition cannot be completely cured, it may turn into a low-grade form of the condition that comes and goes for months or years. This form may occur with bone pain, tenderness and sinus drainage. Because bone infections often block blood vessels, parts of the bone die. This leads to the spread of the infection, and abscesses that drain through the skin may develop.
Causes and Risk Factors
The staphylococcus aureus bacteria from a wound or a contaminated needle usually cause the acute form. A combination of microbes, including tuberculosis, HIV and others that affect immunity usually are responsible for the chronic form. Anyone who has a debilitating disease, cancer or diabetes or has had radiation therapy, dialysis or intravenous drugs has a higher risk of this infection.


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Fundraising for memorial playground tops $100,000
RIVER FALLS, Wis. -- Money to build a playground to memorialize three girls murdered by their father has surpassed $100,000.
That amount will need to multiply fivefold before Tri Angels Playground in River Falls becomes a reality.
Amara, Cecilia and Sophie Schaffhausen were killed by their father, Aaron Schaffhausen, in July 2012. He is serving a life sentence for the crimes.
Becky Stotz, grandmother of the three girls, said collections for the playground remain steady. Stotz's daughter and the girls' mother, Jessica, remarried Nov. 16 and moved to a new house in River Falls.
As of last week, the playground fundraising total was $108,885. The aim is to raise at least $550,000.
Stotz, who lives in Springfield, Ill., said a fundraising committee has applied for a $100,000 grant for the project from Bremer Bank and a $50,000 grant from an unnamed local business.
To donate, visit www.unlimitedplay.org/tri-angels-playground or mail a check to Unlimited Play/Tri-Angels Playground/4140 Old Mill Parkway/St. Peters, MO 63376.


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Emmure Releases Video For 'MDMA' [VIDEO]
Just in time for the 'Trespass America' tour Emmure have released the video for their track 'MDMA' which can bee seen here.
"Slave To The Game" is the 5th studio album from Emmure and I couldn't be more pleased with the album. I have never seen Emmure live but that will change on July 24th when they destroy the stage for the 'Trespass America' tour.
I ahve been a fan of Emmure for such a long time and have featured them several times on Everything Goes Black.
Check out the video for "MDMA" and get ready to Emmure live right here in Lubbock, Texas for the 'Trespass America' tour starring Five Finger Death Punch, Killswitch Engage and many more. If you need more details then check them out right here.
Subscribe to KFMX FM on


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Archive of ' Silly '
13 responses - Posted 04.01.08
So the monkeys over at World of Matticus have caught on that I happen to play on an RP server. And not only that, I play on *the* RP server – yes, you know it – Feathermoon, home of …continue
5 responses - Posted 03.31.08
If a Gnoll in search of augur found a skull inside his lager,
…would it be Hogger's Noggenfogger?
If Hogger had a boggan set atop his hairy noggin, as he augured in his lager called the Hogger Noggenfogger,
… …
3 responses - Posted 03.20.08
This was given to me by Shugotenshi, who was there at its original telling by Ninquelote, on the Feathermoon server. I did not witness the original telling, but I have so loved this poem that I found it needed to …continue
7 responses - Posted 03.08.08
(To the tune of Gloria Gaynor's "I Will Survive")
… continue
At first I was dazed,
Then I was Mortal Striked,
Kept thinking I would always live
If I could duck and hide
But they've made so many mods
For Arathi and


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Area Residents Invited to E-Cycle with Students on Saturday, April 25
Published on Tue, April 21, 2009
Marywood University students under the direction of Dr. Robert Sadowski, will be hosting an e-cycling event Saturday, April 25th from 10:00 AM to 2:00 PM at Marywood University's "Pit" parking lot, located on the Marywood campus across from Loughran Hall.
E-cycling is like recycling, but instead of paper, glass or plastic - e-cycling involves the recycling of electronics, such as computers, televisions, and old cell phones. The electronics get broken down and parts get salvaged.
John Hambros from Apex Waste Management approached Dr. Sadowski's Public Relations: Principals and Practices class four months ago to solicit their help in promoting the event. Apex will be supplying tractor trailers and Gaylord bins to store and haul the recyclables away. Marywood University subsidized some of the promotion costs and Chartwell's Dining Services agreed to provide food and beverages for event volunteers.
The thirteen students in the class, committed to protecting and preserving their environment outside of the classroom, broke up into seven groups to produce Radio and TV promotions and Advertising; T-shirts and Neighborhood PR; Band relations; Print ads and Security; Marketing and Community Relations; web development and Web ads; and signage and directions.
Students from the class feel that the project has helped them to build character. "As a very involved student at Marywood, I constantly strive to live the University's mission of service. This project has allowed me to take the leadership concepts that I have learned in class and use them to advance this project. I think we are all a little impressed by how well we have been able apply what we have learned in the classroom to a life experience." Marywood student, Lindsay Jacob said. "With the tremendous support we have received from the faculty and staff at Marywood as well as our sponsors at Waste Management, we know that this event is going to be a great success" she added.
Local firefighters have volunteered to help lift any heavy equipment out of cars into the tractor trailers to encourage people to participate regardless of the size or weight of their e-cyclables.
It's important to note that the following items cannot be e-cycled:
- Microwaves
- Smoke alarms, Detectors, Fire Alarms
- Thermometers
- Dehumidifiers
- Large Appliances (such as refrigerators and washing machines)
- Medical equipment
- All items that are not dry
- Anything so large that it needs to be put into the back of a large truck
Directions to the drop-off location will be posted all over campus on the day of the event. For further information please visit our website www.greenonproject.com.


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Cammy Bean's Learning Visions
Musings on eLearning and instructional design from Kineo's VP of Learning Design.
Sunday, January 31, 2010
Learning Visions Has Moved
I've moved my blog and switched the feed. Right now I'm just testing to see if the old feed still works if I switch back.
If you'd like to keep reading Learning Visions, I'm still posting away. Just stop by the blog and subscribe to my new feed.
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Rubbing alcohol is a liquid used topically for disinfection or to soothe, cool, or warm the skin! Witch hazel is an astringent which is used to tighten pores & remove excess oil on your skin! They are however very similar in what they can do! ! Any Suggestions here?
The rubbing alcohol, witch hazel and wintergreen can all be purchased at a pharmacy or health Difference Between Rubbing and Polishing Compound Source:http://www.ehow.com/how_2364225_make-rubbing-compound.html
for short- rubbing alcohol is very extremely hazardous to humans and bacteria, and is chemical based. And whitch Hazel is organic and is used for acne cream-cuts-shaving cream etc. rubbing= Isopropyl alcohol- Isopropyl alcohol (also 2-propa
Rubbing Alcohol and Denatured Alcohol RUBBING ALCOHOL is intended for use on the skin only. It was made in the old days for rubbing on the skin, on a inflammed Joint. It was also Rubbed on skin to reduce fevers. It is no longer used much fo… Source:http://wiki.answers.com/Q/What_is_the_difference_between_denatured_and_rubbing_alcohol
Difference In Grain and Rubbing Alcohol If memory serves me correctly, grain alcohol is derived from the fermentation of various grains, with the most common being from corn, and it's safe to consume. On the other hand, rubbing alcohol cont… Source:http://wiki.answers.com/Q/What_is_the_difference_between_grain_alcohol_and_rubbing_alcohol
Rubbing alcohol is most often used for disinfecting purposes, such as cleaning wounds. But the common household product is handy for a number of less obvious tasks. Keep a bottle on hand to get out of some sticky situations. Source:http://www.ehow.com/how_5740030_use-rubbing-alcohol-different-ways.html
Recipe for astringent calls for distilled witch hazel. Should I use witch hazel with alcohol or not? Does the distilled mean one or the other? Which works better? Thanks I mea
Witch Hazel has newt liver and wizard piss
for short- rubbing alcohol is very extremely hazardous to humans and bacteria, and is chemical based. And whitch Hazel is organic and is used for acne
Spamer is not welcome,every link should be moderated.


text:
Product Summary
CE-VG0112-S1 Video Console/Extender extends VGA video and stereo audio transmission over one economical CAT5/5e/6 cable. Supports 2-channel stereo audio. Supports VGA, SVGA, XGA, SXGA and multisync monitors.
General Information
|Product Type
|Video Extender/Console
|Manufacturer Part Number
|Manufacturer Website Address
|SIIG, Inc
|Product Model
|Product Name
|CE-VG0112-S1 Video Console/Extender
|Brand Name
Technical Information
|Graphics Resolution
|1920 x 1200
|Operating Range
|1080 ft @ 800 x 600
|246 ft @ 1920 x 1200
|Connectivity Media
|Category 5
|Category 5e
|Category 6
|Number of Input Devices
|Number of Output Devices
|Graphics Modes
|Other Ports
|1 x USB
|1 x 3.5mm Male Audio
|1 x 3.5mm Female Audio
|Input Ports
|1 x 15-pin HD-15 Male Video Transmitter
|Output Ports
|1 x 15-pin HD-15 Male Video Receiver
|Package Contents
|Standard Warranty
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Russell Martin is reportedly seeking a four-year deal worth between $9 million and $10 million annually, according to Andrew Marchand of ESPNNewYork.com. Marchand doesn't think the Yankees would be willing to re-sign Martin under those terms.
Martin has been highly coveted this offseason. CBSSports.com's Jon Heyman reported the Rangers had Martin on their wish list, and that the Pirates, Mariners and potentially the White Sox could all be looking to add catching.
For more on this story visit: Chris Cwik, CBSSports.com


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On Tennis: Nadal's finale in Melbourne brings Federer to tears
Published 10:00 pm, Sunday, February 1, 2009
Three years ago Roger Federer wept in the arms of Rod Laver as he accepted the winner's trophy at the Australian Open. There were more tears this week Down Under, with the Swiss not too happy this time.
How could he be, after dropping yet another five-set thriller to nemesis Rafael Nadal? Federer must have thought he had Nadal right where he wanted him, given the bullish Spaniard spent more than five hours on court in his captivating semifinal against another Spaniard, Fernando Verdasco, two days earlier.
Nadal's 7-5, 3-6, 7-6 (3), 3-6, 6-2 win in 4 1/2 hours denied Federer the historic 14th Grand Slam and has to be an even bigger blow to the world No. 2 than last July's Wimbledon defeat that ended 9-7 in the fifth. Federer sobbed during the trophy presentation, Laver nearby, receiving consolation from none other than Nadal, who acted like a big brother.
"In the first moment you're disappointed, you're shocked, you're sad, then all of a sudden it overwhelms you," Federer said. "The problem is, you can't go in the locker room and take it easy and take a cold shower. You can't. It's the worst feeling."
The stars seemed aligned for Federer. He landed on the gentler side of the draw, avoided defending champion Novak Djokovic in the semifinals and had an extra day of rest between the semi and final.
But Nadal, not picked by many to win the title because of ongoing knee problems and Andy Murray's emergence, is a different specimen. He rallied from 4-2 down in the first set and thwarted all six break points in the third. Indeed, winning the third was the turning point. (Curiously, Federer actually won more points altogether, 174-173.)
Federer's serve always seems to malfunction against the scurrying Mallorcan, and it happened again: The first serve percentage stood at 51.
Federer felt he played a "terrible" fifth set -- Nadal didn't. Like at Wimbledon, Nadal protected his serve gamely in the decider, conceding a miserly three points.
It's now six Grand Slams for Nadal at the tender age of 22. He's a U.S. Open away from completing his Grand Slam collection.
He's the undisputed No. 1 and will be for a while.
SIZZLING SERENA: How dominant was Serena Williams in the women's final? Here are some stats that prove the point: Her unfortunate victim, the affable Dinara Safina, won only eight points the first set, and in the entire match, Safina triumphed on six points behind her own second serve.
The 6-0, 6-3 rout was so inevitable that journalists began wondering what the shortest finals in Grand Slam history were. Safina lasted 59 minutes, by the way, not close to any records.
The tennis gods were on Williams' side throughout the fortnight, too. Trailing rising star Victoria Azarenka by a set in the fourth round, the Belorussian quit in the middle of the second, succumbing to a virus. In the quarterfinals, the always-jittery Svetlana Kuznetsova couldn't finish Williams off, failing to serve out the encounter in the second set.
As usual, Williams picked up her game when it really mattered, steamrolling Elena Dementieva, the hottest player on the tour, in the semis before dispatching Safina.
After claiming the U.S. Open, a second Serena Slam is very much a possibility. Williams collected major No. 10, three more than older sister Venus.
"Well, I wanted to get to 10," Williams said.
Less important to Williams, she regained the No. 1 ranking from the disappointing Jelena Jankovic.
NOT AGAIN: Australian Open organizers had a tough fortnight. A riot broke out between rival fans, a streaker invaded a doubles match involving the Williams sisters, the extreme heat policy was called into question, and scheduling turned out be a major gripe, especially for Djokovic.
Officials blundered by making Djokovic's quarterfinal tilt with Andy Roddick an afternoon affair a round after the Serb finished extremely late. To boot, the pick of the day's matches usually features in the evening, and it didn't take a genius to figure out Federer's encounter against Argentinean Juan Martin Del Potro, which ended up being played at night, would be a dud. Djokovic and Roddick, meanwhile, had ample history following their ill-tempered meeting at the U.S. Open.
While he got shafted by the schedule, there's no excuse for Djokovic retiring -- again -- in a big match. He quit in the fourth set against Roddick, undone by the oppressive heat. Roddick didn't appear bothered by the 105-degree temperatures one bit.
A-ROD'S OZ: Roddick can't be disappointed with his exertions Down Under. He put away foes he should have -- and then got Federer. Federer reads Roddick's bullet serve well, and it's no contest when the two engage in baseline rallies.
Roddick trails Federer 16-2 in their head-to-heads, and to get a third victory he needs the latter to have one big off-day, as was the case in Miami last year.
Perhaps they'll meet in the first round of the Davis Cup world group in March, when the U.S. hosts Switzerland in Birmingham, Ala..
"Overall I think it was a good event," Roddick said. "There's a whole laundry list of positives."
SECOND SERVES: Out injured, Maria Sharapova dropped out of the top 10 for the first time since 2004. ... Federer advanced to his 18th Grand Slam final, one shy of Ivan Lendl's men's record.


text: While there were no 'formal' discussions about the possibility of moving to an 8-game football schedule at the recent Pac-12 meetings according to Utah athletic director Chris Hill, the topic was certainly bantered about among the coaches, whose ultimate desire is to have a consistent format for all conferences. As for Hill, he said he prefers keeping the 9-game schedule. "I like playing as many Pac-12 teams as possible," he said. One of the reasons he feels it is important to do so is for strength of schedule, a factor that will become a bigger player once college football employs its new post-season format. "We feel that is why the Big Ten is going to nine games (in 2016), because they need that strength of schedule," he said. Other topics that were discussed was the men's basketball tournament which was held in Las Vegas at the MGM Grand in the first stint of a three-year deal. The tournament was just one of four, along with the Mountain West, West Coast and Western Athletic conferences, that the city hosted but the first for the MGM Grand's Garden Arena. From what Hill said, the first time went over well. "People really liked the location, atmosphere and enthusiasm around it," he said. The other topic discussed was what teams can do about concussions. Hill said he feels strongly about being proactive and wishes for more research on how college football can become safer. "I would welcome more input and support it," he said. "We need to do as much as we can. It's a topic we can't ignore and has to be a top priority." - Lya Wodraska