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The Role of Intralesional Therapies in Melanoma
The Role of Intralesional Therapies in Melanoma
The US Food and Drug Administration has been rapidly approving new checkpoint inhibitors and targeted therapies for melanoma and other tumors. Recently, it approved the first intralesional therapy, talimogene laherparepvec (T-VEC), for the treatment of metastatic melanoma lesions in the skin and lymph nodes. Several other intralesional therapies (PV-10, interleukin-12 electroporation, coxsackievirus A21 [CVA21]) are entering later-stage testing. Locally injected agents have clearly shown their ability to produce local responses that can be durable. The possibility that they also stimulate a regional and even systemic immune response is exciting, as this potential effect may have utility in combination regimens; such regimens are an area of active research. Favorable responses with minimal toxicities in monotherapy trials have led to the first melanoma studies of T-VEC in combination with the cytotoxic T-lymphocyte–associated antigen 4 inhibitor ipilimumab and, separately, with the programmed death 1–blocking antibody pembrolizumab. Studies of PV-10 with pembrolizumab and of CVA21 with pembrolizumab are also being initiated. Preliminary analyses of the results of the first combination trials, which show higher response rates than with either agent alone, offer some optimism that these locoregional therapies will find application—as treatment for patients who cannot tolerate systemic immunotherapies, to alleviate locoregional morbidity, and perhaps even to "prime" the immune system.
Melanoma accounts for about 4.5% of new cancers (21.6% per 100,000) and 1.7% of all cancer deaths in the United States. The incidence of melanoma has been gradually increasing for several decades, with the annual rise averaging 1.4% per year over the last 10 years. It is estimated that the number of deaths from melanoma approached 10,000 in 2015.[1]
After frustrating decades of little progress, the landscape of melanoma therapy was dramatically altered with the first report in 2010 of significantly improved overall survival with the cytotoxic T-lymphocyte–associated antigen 4 (CTLA-4) inhibitor ipilimumab, compared with a vaccine, and subsequently, compared with the then "standard" chemotherapeutic agent dacarbazine (DTIC). Following the US Food and Drug Administration (FDA) approval of ipilimumab the following year, approvals of other targeted therapies (against BRAF and MEK), and most recently, approvals and extended indications for other checkpoint inhibitors, especially the programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1) inhibitors, have constituted a revolution characterized by higher response rates and the potential for extended survival. Clearly, we are now in an era where new treatments are showing positive results and are becoming available at an unprecedented rate.
Why an Interest in Locoregional Therapies?
Direct treatment of cutaneous melanoma lesions is on a separate track from that of systemic therapies, with its rapid advances. However, interest in strategies focused on the direct treatment of cutaneous melanoma lesions has its own pedigree. Melanoma is unique in its propensity to produce cutaneous, subcutaneous, and nodal metastases. This unique pattern of metastatic spread affords us an opportunity to access the tumor and manipulate it for potential therapeutic benefit. Furthermore, while it is true that metastatic melanoma is a systemic disease—and also that mortality is not caused by dermal or subcutaneous lesions but rather by metastases to the liver, brain, lung, and other sites—locoregional disease is often disfiguring and painful, and locoregional therapy may provide clinical benefit.
A particular challenge is posed by the in-transit metastases that develop in 6% to 10% of patients with primary melanomas > 1.0 mm in Breslow thickness.[2] In some patients, in-transit metastatic disease may remain indolent and regionally confined for years or decades. In these patients, clinical results with systemic therapy have been less than optimal, and locoregional therapeutic options, such as limb perfusion and intralesional treatments, have thus been explored.
Intralesional therapy for melanoma has a long history. The unique relationship of melanoma to the immune system has stimulated research in this area for many years. Murine experiments showed heightened host immune responses against transplanted experimental tumors after inoculation with bacillus Calmette-Guérin (BCG).[3,4] Dummer et al demonstrated BCG-induced increases in melan-A–specific cytotoxic T cells.[5] Morton et al, in 1974, reported 7 years' experience with 151 malignant melanoma patients who received BCG immunotherapy alone or as an adjunct to surgery.[6] In lesions limited to skin, regression was 90% in injected lesions and 17% in uninjected lesions, with one-fourth of patients remaining disease-free for 1 to 6 years.
Enthusiasm for intralesional BCG therapy waned following reports of anaphylactic reactions; disseminated BCG mortality[7]; and, in our own research, the occurrence of punctate abscesses in more than two-thirds of treated patients, as well as failure to impact outcomes.[8] Other agents of mostly historical interest include intralesional interleukin (IL), interferon (IFN), and granulocyte-macrophage colony-stimulating factor (GM-CSF), which have not shown consistent or durable efficacy and have not been rigorously tested in randomized trials. Here we review agents that have been evaluated or are currently undergoing evaluation in the modern era of melanoma therapeutics.
Velimogene Aliplasmid
Velimogene aliplasmid (Allovectin-7), a plasmid/lipid complex with human leukocyte antigen (HLA)-B7 and β2 microglobulin DNA encoding sequences, was granted orphan drug designation by the FDA back in 1999. HLA-B7 and β2 microglobulin are components of major histocompatibility complex class I (MHC-I), expression of which is often altered during cancer progression, allowing tumor cells to evade the immune system.[9] A role in augmenting the immune system's ability to recognize and target melanoma cells was hypothesized because velimogene aliplasmid increases HLA-B7 cytotoxic T-cell frequency fivefold, upregulates/restores MHC-I molecules, and induces a proinflammatory response.
The agent showed promise in a phase II trial, which was conducted in 133 patients with stage IIIB/C and IV M1a/b injectable cutaneous, subcutaneous, or nodal melanoma lesions.[10] Complete responses (CRs) and partial responses (PRs) were reported in 3.2% and 8.7% of patients, respectively, with stable disease (SD) in 25% of the two patient cohorts combined. Time to death was significantly longer in responders than in nonresponders (P = .036). Of the patients with stage IV M1a/b disease, 21% (9/42) had responses in noninjected target lesions. However, the subsequent phase III Allovectin Immunotherapy for Metastatic Melanoma (AIMM) trial failed to show benefit.[11] Among 390 patients with stage IIIB/IV M1a/b melanoma randomized 2:1 to velimogene aliplasmid or to intravenous DTIC or oral temozolomide (TMZ), the primary endpoint of response rate at ≥ 24 weeks was lower in the velimogene aliplasmid group at 4.6%, compared with 12.3% for DTIC/TMZ (P = .010). While the duration of response in the velimogene aliplasmid responders was marginally longer than with DTIC/TMZ (P = .066), overall survival was shorter (median, 18.8 months [95% CI, 16.6–21.3 months] vs 24.1 months [95% CI, 17.1–27.9 months]; P = .491). The velimogene aliplasmid program was discontinued.
Electroporation With Plasmid IL-12
With intratumoral electroporation, tumor cell pores are opened when a mild electrical current is introduced directly through an electrode. The intention is to thereby facilitate greater influx of a cytotoxic agent over a longer period of time than would occur with systemic administration of the same agent. Achieving high levels of IL-12 protein expression stimulates a local proinflammatory process, which leads to a targeted immune response; enhances the immune capacity of natural killer (NK) and T cells; and upregulates IFNγ, as well as antigen presentation and processing. With reduced systemic drug concentrations, side effects are minimized. This feature is of particular value with respect to IL-12, a cytokine that is toxic when given systemically.
Interim phase II results[12] of first treatment with electroporation of IL-12 in 28 patients with advanced melanoma, after 24 weeks of treatment, reported at the American Society of Clinical Oncology 2014 Annual Meeting, revealed a 32.2% objective response rate (ORR; the primary endpoint), with a CR in 10.7%. Among lesion responses (n = 85), the CR rate was 45%, and the PR rate was 8%, with SD in 31%. Responses were observed in untreated lesions in 59.1% of evaluable patients (13/22).
The electroporation treatment produced no toxicity and no serious adverse events, aside from injection site pain (69.0%) and inflammation (20.7%). Grade 3 pain was reported in 1 patient.
An exploratory analysis showed that intratumoral NK cells doubled from pretreatment through day 11, and doubled again by day 39, with an increase in the frequency of activated circulating NK cells. In the phase II trial, a maximum of four treatment cycles at 12-week intervals were allowed. A planned expansion protocol in melanoma patients will evaluate increased treatment frequency.
In October 2015—around the same time that the FDA was granting several approvals and indication extensions to checkpoint inhibitors—the agency approved the oncolytic virus (herpes simplex virus 1)–derived therapy talimogene laherparepvec (T-VEC) for the treatment of melanoma lesions in the skin and lymph nodes. T-VEC encodes GM-CSF, and is thought to replicate in tumor cells, lysing cells in injected tumors. Antigen-presenting cells then take up the lysed cells. Local expression of GM-CSF may also evoke an enhanced adaptive antimelanoma response (Figure).
T-VEC was compared with GM-CSF in the phase III OPTiM study in 436 stage IIIB/C and IV melanoma patients who had injectable and unresectable disease.[13] Durable response (complete or partial), the primary endpoint, was defined as a response lasting continuously for at least 6 months and begun within 12 months of initiation of therapy. Patients were randomized 2:1 (295:141) to intralesional T-VEC (initially, ≤ 4 mL × 106 pfu/mL; then after 3 weeks, ≤ 4 mL × 108 pfu/mL every 2 weeks) or subcutaneous GM-CSF (125 µg/m2 daily × 14 days every 28 days).
The durable response rate (DRR) in the intention-to-treat analysis was 2.1% in the GM-CSF arm and 16.3% in the T-VEC arm, a treatment difference of 14.1% (95% CI, 8.2%–19.2%; P < .0001). The ORR in the 141 patients who received GM-CSF was 5.7% (95% CI, 1.9%–9.5%). It was 26.4% in the 295 patients who received T-VEC (95% CI, 21.4%–31.5%; P < .0001), for a treatment difference of 20.8% (95% CI, 14.4%–27.1%; P < .0001). In the T-VEC responders, the CR rate was 41% (10.8% among all patients who received T-VEC; 0.7% for all patients who received GM-CSF). PR rates were 15.6% and 5.0%, respectively, for the entire T-VEC and GM-CSF arms.
In the stage IIIB/C melanoma patients with no disease spread to distant organs, the DRR was 33% for T-VEC compared with 0% for GM-CSF. Differences in DRRs between groups were smaller for stage IV patients (11% for T-VEC, 7% for GM-CSF).
With regard to median overall survival (OS), a secondary endpoint, T-VEC had a 4.4-month advantage over GM-CSF (23.3 months vs 18.9 months [hazard ratio (HR), 0.79 (95% CI, 0.62–1.00); P = .051]); this approached but did not achieve statistical significance.
The OPTiM investigators, in an analysis of lesion-level responses, reported that among 2,116 lesions in patients treated with T-VEC, tumor area decreases of ≥ 50% occurred in 64% of injected lesions, in 34% of uninjected nonvisceral lesions, and in 15% of uninjected visceral lesions.[14]
The most common adverse event was fatigue (occurring in 50.3% of patients in the T-VEC arm, and in 36.2% of those in the GM-CSF arm), with chills (48.6% in the T-VEC arm; 8.7% in the GM-CSF arm) and pyrexia (42.8% in the T-VEC arm; 8.7% in the GM-CSF arm) the next most common. The only grade 3/4 event that occurred in ≥ 2% of patients was cellulitis (seen in 2.1% of the patients who received T-VEC).


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- Photography
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- Surveillance
- Optics
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- Used Dept
Here's the situation: you buy a brand new Mac with an empty 500GB hard drive. Your old computer has a smaller hard drive, but it's completely full and not very organized. You can use software that will transplant the chaotic mess from your old drive directly into your new computer, or you can take your time and manually move files over, organizing as you go. In this post I'll share some tips on how you can turn a new leaf when you get a new computer.
A lot of the information presented in this post can also be applied to PC users. First, here's a little back story. In early 2004, I bought my first Macintosh computer. After six years of dependable service I finally decided to upgrade to a new MacBook Pro with an i5 processor. My old dual 1.8 gHz G5 was still working rather well, but the outdated PPC processors weren't really up to snuff with the latest Mac OS, and I was encountering more and more annoying little problems with software drivers and Internet applications.

subset: commoncrawl


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It's taken a while, but I've finally replaced my yucky old DYI Yahoo! Site Builder website with something a little more professional looking. Happy to report that as of this week www.sarabakescakes.com is up and running! Little kinks still to be worked out, including resizing many of the photos (what a pain in the butt!) and going through Flickr and adding more, but so far so good.
Doug won a free website design from a great company called SiteDesignWorks and he gave it to me. I was able to choose from a number of predesigned sites - ended up choosing one called Prismatic. They even designed a logo for me.
Is it my dream website for Sara Bakes Cakes? No. But is it a 1,000 times better than the old one? Absolutely. Check it out!


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Overall a pleasant stay. We were overnighting in Cairns in order to catch an early morning flight. This property represents good value given its location close to the esplanade area. The property is probably showing its age, but has all you need and is clean and tidy. The rooms are very spacious , with a balcony allowing views of the sea. The bed was the best we had experienced in our travels. The check in was efficient and well organised as was the check out. All in all just what you want from a 4 star corporate city hotel, efficiency and value.
- Official Description (provided by the hotel):
- Oceanfront 12-story hotel towers, 3 blocks from city center. ... more less
- Reservation Options:
- TripAdvisor is proud to partner with Booking.com, Expedia, Ctrip TA, Travelocity, Hotels.com, Orbitz, Agoda, TripOnline SA, Hotwire, Cancelon, HotelQuickly and getaroom.com so you can book your Rydges Esplanade Resort Cairns reservations with confidence. We help millions of travelers each month to find the perfect hotel for both vacation and business trips, always with the best discounts and special offers.
- Also Known As:
- Rydges Esplanade Resort Cairns Hotel Cairns
- Rydges Esplanade Hotel Cairns
- Rydges Esplanade Cairns
- Rydges Cairns


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eHere is my made up season of iCarly. It's called iCarly M.U.S (a.k.a., Made Up Season). Here's the season's list of episodes:
- iGet a Concert
- iKiss a Superstar
- iOpen Sam's Secret Jar
- iAm in a Band
- iFind Gibby a Girlfriend
- iFind Out Shelby Marx and Wade Collins are Dating
- iMake My Own Website
- iWill Undo It (Seddie episode)
- iThrow Spencer a Huge Birthday
- iOpen an iCarly Pie Shop
- iCafe
- iMake Sculptures
- iAm NOT Melanie
- iMake am in a Movie
I put some episodes with Carrie Underwood songs (example: iWill Undo It). I hope these episodes are good to you! I will write iGet a Concert later! sorry if i'm copying you (if you're doing it too!). I got this idea from Lotstar. Enjoy! :D


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What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
What if I had reached a maximum submition in a month?Asked by UjangSutresno on November 21, 2015 at 02:15 PM
What if I had reached a maximum submition in a month? can I reset this? Thanks
This is a re-post of a comment on Adding a Feedback Button to Blogger
JotForm Support
There is an automatic account submission limits resets that occrus every 1st Day of the Month.
In case you have exceeded your account limits before the Reset day occurs, your account will go into Overlimit Status and forms will be temporarily disabled.
Disabled forms will not be able to receive submissions and will not load as well.
To continue using the forms and remove disable status, either upgrade the account to the next higher plan (for expanded limits), or wait for the auto reset to occur.
Hope this help. Let us know if you require further assistance.


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This week, the House passed H.R. 436, the Protect Medical Innovation Act. The legislation focused on three issues affecting the health care industry. First, it repeals the medical device tax established under ObamaCare that will take effect next year. The 2.3 percent tax will be imposed on the sale of medical devices by manufacturers or importers. According to an industry study, up to 43,000 American jobs could be eliminated due to this new tax.
"It seems like every day we find a new problem within ObamaCare," stated Westmoreland. "There are new taxes, unsustainable programs even the White House admits won't work, and more bureaucratic red tape than I have ever seen in my lifetime. That's why I strongly support fully repealing this destructive law and, like many Americans, am awaiting the decision by the Supreme Court at the end of this month on the constitutionality of it. In the mean time, I'm going to continue to work with my colleagues here in the House to repeal as many of the harmful aspects of ObamaCare as we can."
H.R. 436 also contains provisions dealing with health-related savings accounts. Under ObamaCare, Americans may not currently use tax-free distributions in their health-related savings accounts to purchase over-the-counter medications without a prescription from a physician. Over-the-counter medications are often just as important -- and as expensive -- as prescription medications and should be an acceptable expense for health-related savings accounts. H.R. 436 fixes this provision of ObamaCare. It also adjusts current law to allow Americans to keep unused flexible spending accounts (FSAs) up to $500 at the end of each year. The unused money would be included as taxable income for the employee.
"These health-related savings accounts that many employers offer are a great way for American families to pay for their health care expenses," stated Westmoreland. "With ObamaCare making health care and health insurance even more expensive, these tax-free savings accounts provide a much needed relief. H.R. 436 makes these small changes to the rules for health-related savings accounts so they are able to be a useful tool for all Americans."
The legislation passed the House with large bipartisan support by a vote of 270-146 and was sent to the Senate. It is unclear at this time if or when the Senate will act on it.
Source: http://westmoreland.house.gov/index.php?option=com_content&view=article&id=659&Itemid=363


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I may have overbaked this one a little but after over an hour it still wasn't done. Now the crust is a bit crunchy but inside it's still tight-crumbed and tender, a pefect combination. I used pecans instead of walnuts -must make a mental note to avoid using nuts altogether as Maya hates them -roll eyes. Use chocolate chips next time to make it even more kid-friendly.
Mark Bittman's ultimate banana bread
(adapted from here, where you'll find the recipe in cups)
Note to self : although this recipe is excellent, I'll omit the 50g white sugar next time as, upon maturing, the bread is quite sweet, not a problem in itself but spread Nutella on top...
110g butter
180g plain flour
60g wholewheat flour
fat pinch of salt
1 1/2 tsp baking powder
100g golden caster sugar
1 sachet vanilla sugar (or 1 tsp vanilla extract)
50g white sugar
2 eggs
3 mashed bananas
1/2 cup chopped nuts
1/2 cup grated coconut


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By Jerry Shriver, USA TODAY
USA TODAY's Jerry Shriver asks top TV chefs....What is the most romantic dining-out experience you've ever had or have dreamed of having?
Pat and Gina Neely
The husband/wife team are part of a family that owns several barbecue restaurants and concessions in Tennessee. They will launch their first Food Network series, Down Home With the Neelys, Saturday (11 a.m. ET/PT).
"My favorite romantic dining experience was when Pat and I went to The Cloister at Sea Island, Ga., in November last year. They brought a table out onto the deck of the room we were staying in, and we had a candlelit dinner overlooking the beach. You could hear the waves, and the dinner was delicious. The most memorable dish was the grilled crab with melted butter. We had on our bathrobes and were barefoot, and the breeze was blowing, but just enough to keep you cool. It was our very first time there and we'll definitely go back."—Gina Neely
"The Cloister is a place of paradise. We didn't anticipate this fantastic romantic dinner because we had just flown in and said we'll just do room service. We took a shower, and then here came these guys with handheld trays. They rolled in a full table on wheels and asked if we wanted dinner in the room or on the patio. The food was outstanding — I had a filet mignon, potatoes and vegetables. But sitting on the deck of the room overlooking the ocean with the sun setting, having a glass of Champagne — it was like the honeymoon all over again." —Pat Neely
Alton Brown
The star of Good Eats (Mondays, 8 p.m. ET/PT) and commentator on Iron Chef America (Sundays, 9 p.m. ET/PT) trained at the New England Culinary Institute and won a James Beard award for his reference work, I'm Just Here for the Food. In March, he'll release Feasting on Asphalt: The River Run, a road journal with recipes.
"It was 1992 and my soon-to-be wife and I were broke, which meant it was the perfect time to go overseas. We flew (had to share a single coach seat) to London, rented a car the size of a microwave and headed south. Along the way we bought some Cadbury chocolate, a pear, a small hunk of cheddar at some roadside stand, and a big bottle of dark beer. After driving through a deluge in southern Dorset we stopped by a field overlooking the sea. We got out our pathetic little bag of goods and picnicked on the grass. The pear was perfect, the cheese was perfect, the beer was … well, you get the point. And then the show began: The sun came out and suddenly hundreds, maybe thousands, of rabbits came up out of their burrows and started hopping around the field. I'd never seen anything like it. It was like a surrealist painting, only tasty. I'll never forget that.
"As for the most romantic meal I'd like to have, I have but two words: Orient Express."
Ingrid Hoffmann
The Colombia native is host of Simply Delicioso (Saturdays, 1:30 p.m. ET/PT), which is shot in her hometown of Miami Beach. She has a book hitting stores Tuesday: Simply Delicioso: A Collection of Everyday Recipes With a Latin Twist.
"I dream of going back to a place I visited with my family about four years ago. It was the perfect setting, and I said I would one day come back with someone special. It's on the edge of Lake Titicaca in Bolivia, in an area called Copacabana, at La Orilla restaurant at the side of the lake. We were there for lunch, which is served on this terrace with this incredible view. It is one of those places touched by God, with strong energy. You feel different. It is not a posh place, but it is where the earth and good food and ambience come together.
"You don't need a lot of candles there. I was served an incredible lake trout, which was served with delicious warm quinoa, and it became the inspiration for the quinoa citrus salad that's in my book.
"If I had to pick a romantic city, it would be Paris and Rome, but being Latin I would add Cartagena (Colombia), one of the most beautiful cities in Latin America because of its well-preserved colonial elements."
Conversation guidelines: USA TODAY welcomes your thoughts, stories and information related to this article. Please stay on topic and be respectful of others. Keep the conversation appropriate for interested readers across the map.


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Nuclear Factor kappa B is central to Marek's Disease herpesvirus induced neoplastic transformation of CD30 expressing lymphocytes in-vivo
Empirical systems biology
Marek's Disease (MD) is a hyperproliferative, lymphomatous, neoplastic disease of chickens caused by the oncogenic
Our results show that a) CD30lo lymphocytes are pre-neoplastic precursors and not merely reactive lymphocytes; b) multiple transformation mechanisms exist and are potentially controlled by Meq; c) Meq can drive a feed-forward cycle that induces CD30 transcription, increases CD30 signaling which activates NF-κB, and, in turn, increases Meq transcription; d) Meq transcriptional repression or activation of the CD30 promoter generally correlates with polymorphisms in the CD30 promoter distinguishing MD-lymphoma resistant and susceptible chicken genotypes e) MDV oncoprotein Meq interacts with proteins involved in physiological processes central to lymphomagenesis.
In the context of the MD lymphoma microenvironment (and potentially in other CD30hi lymphomas as well), our results show that the neoplastic transformation is a continuum and the non-neoplastic cells are actually pre-neoplastic precursor cells and not merely immune bystanders. We also show that NF-κB is a central player in MDV induced neoplastic transformation of CD30-expressing lymphocytes in vivo. Our results provide insights into molecular mechanisms of neoplastic transformation in MD specifically and also herpesvirus induced lymphoma in general.
Lymphomas are the 6th leading cause of cancer mortality in the USA especially in patients younger than 40 years
CD30 overexpression is an evolutionarily conserved process in neoplastic transformation in human and chicken lymphomas of different etiologies
CD30 signaling activates the transcription factor Nuclear Factor-kappa B (NF-κB), which regulates genes associated with cell survival, proliferation, programmed cell death (PCD), stress and immunity
MD transformed lymphocytes have increased MDV oncogene "Meq" expression
Here we show, using MD lymphocytes isolated directly ex vivo that they are either neoplastically transformed and express high levels of CD30 (CD30hi) or are non-transformed and express low levels of CD30 (CD30lo) that: 1) neoplastic transformation is a continuum and the CD30lo lymphocytes within the tumor microenvironment are pre-neoplastic; 2) as the lymphocytes become more neoplastically transformed they become more immune-evasive; 3) the MDV oncogene Meq, has a direct role in this process and 4) NF-κB has a central role in this neoplastic transformation. In vitro
Results and discussion
Because the proteome directly affects phenotype, but the transcriptome merely influences the proteome and thus may only indirectly affect the phenotype
Additional file 1
Additional file 1 contains four worksheets, sheet 1 (CD30 hi Vs. CD30 lo protein expression): contains proteins identified from CD30 lo and CD30 hi cells, worksheet 2 (ChIP identified proteins): contains proteins identified in ChIP experiment. Worksheet 3 (NF κB isoforms subcellular locations): contains actual numbers of peptides of different isoforms of NF-κB in nuclear and extranuclear fractions, and worksheet 4 (Phosphorylated peptides locations) contains actual numbers of phosphorylated and non-phosphorylated peptides identified. (XLS 1901 kb)
Click here for file
Functional modeling
To visualize the differences between the CD30hi and CD30lo lymphocytes proteomes (Figure
Figure 1
Proteomics based network model showing differential expression of proteins in canonical cancer pathways and the intricate involvement of Meq, via hypothesized CD30-NF-κB-Meq pathway
Proteomics based network model showing differential expression of proteins in canonical cancer pathways and the intricate involvement of Meq, via hypothesized CD30-NF-κB-Meq pathway. Canonical Network model of proteome data using differential proteomic expression (CD30hi compared to CD30lo) mapped against cancer specific KEGG pathway "Pathways in cancer" and further modified by adding published Meq interaction and hypothesized Meq-CD30-NF-κB-Meq pathway. Light black solid lines indicate direct interaction, dotted lines indicate indirect interaction. Red dotted lines indicate our hypothesized CD30-NF-κB-Meq pathway. Literature based Meq interacting proteins are indicated by dark black lines originating from Meq.
A mixed pattern emerged with protein levels increasing, decreasing and not changing. However, in several of the pathways described below, key regulatory proteins were differentially expressed: NF-κB, IKK, VEGF, MDM2, CD30, HSPA2, MYC, JUN, TGFβ, and Meq were increased; whereas, RB, PENK, and BRCA2 were decreased. This indicates that neoplastic transformation is being regulated by these key proteins. The MDV oncoprotein Meq interactions, and our hypothesized Meq-CD30-NF-κB feed-forward loop, suggest that Meq interacts with several key proteins involved in neoplastic transformation, immune evasion and cell survival.
Ingenuity Pathway Analysis (
To this point our modeling was on a global scale. Using the same data (Additional file
Growth factor receptors activate the MAPK, JAK-STAT, and, through PI3K/AKT, the MTOR signaling pathways. The MAPK pathway activates JUN, FOS and MYC, and the JAK-STAT pathway activates VEGF and both promote proliferation and angiogenesis. In the MAPK pathway, HRAS was decreased and JUN and MYC were increased. JUN mRNA was decreased and, as JUN transcription is autoregulated by JUN protein (45), and JUN heterodimerizes with Meq. We suggest that even though total JUN protein was increased in CD30hi lymphocytes, it is not available for auto-transactivation, an alternative possibility is that as JUN protein is stabilized by post-translational interactions with Meq, the JUN mRNA may not actually reflect the total JUN protein levels
Activated PI3K phosphorylates AKT, which in turn activates IKKA, MTOR (FRAP1) and MDM2 and inhibits FKHR, CASP9, BAD, p27 and p21 genes
d) A
Biological processes associated with neoplastic transformation and immune-evasion
At a higher level, the
Figure 2
Proteomics based Gene ontology (GO) based quantitative modeling of cellular immunophenotypes of CD30lo and CD30hi cells
Proteomics based Gene ontology (GO) based quantitative modeling of cellular immunophenotypes of CD30loand CD30hicells. Quantitative modeling of the CD3lo and CD30hi lymphocyte proteome by Gene Ontology Biological Processes (A) and immunophenotype using
Next, we compared the CD30hi and CD30lo lymphoma cell immune-phenotypes
Transcriptional regulation
To identify potential direct transcriptional proteome regulation, we used the 44 K Agilent chicken microarray
Additional file 2
Additional file 2 contains three worksheets, sheet 1 contains the results from the statistical analysis of microarray data, sheet 2 contains expression data for only miRNAs and sheet 3 contains predicted targets genes and the relative protein expression of the targets of the miRNAs. (XLS 8422 kb)
Click here for file
Additional file 3
Additional file 3 contains mRNA and protein expression fold changes in terms of CD30 hi over CD30 lo cells. (XLS 859 kb)
Click here for file
Figure 3
mRNA and protein correlation and GO based modeling of 88 concordantly expressed host gene products
mRNA and protein correlation and GO based modeling of 88 concordantly expressed host gene products. Overall correlation (
Specific GO-based BP modeling of these 88 concordantly-expressed genes shows that they are involved in BPs known to be perturbed in, and central to, neoplastic transformation (Figure
We next ranked the genes based on their mRNA: protein expression correlation, and then grouped them into pentiles and compared the distribution of BP by pentile (Figure
We next identified the numbers of putative canonical MDV Meq binding sites (searched for the core AP-1, MERE I and MERE II motifs as described in
Additional file 4
Additional file 4 contains pentile wise Meq binding sites for 88 concordantly expressed host host gene products. (XLS 30 kb)
Click here for file
miRNAs are non-coding post transcriptional repressors potentially important in neoplasia and we identified 152 expressed chicken miRNAs (Additional file
We used the
CD30hi lymphocytes have increased levels of activated NF-κB
Constitutive NF-κB activation is a proposed mechanism by which overexpressed CD30 induces neoplastic transformation in human HL and NHL and in MD
We found that NF-κB p50 (functional NF-κB1), p65 (RelA) and RelB and IKKα proteins all increased in CD30hi lymphocytes (Figure
Figure 4
Schematic diagram of the hypothesized Meq-CD30- NF-κB feed forward loop, subcellular localization and phosphorylation status of different isoforms of NF-κB
Schematic diagram of the hypothesized Meq-CD30- NF-κB feed forward loop, subcellular localization and phosphorylation status of different isoforms of NF-κB. Schematic diagram of hypothesized Meq-CD30-NF-κB feed forward loop and showing differential expression of NF-κB isoforms (A). Comparison of the amount and sub cellular localization of NF-κB isoforms: circle size proportionately portrays relative protein amount; equal size indicates no differential expression at P < 0.05 (B). In CD30hi lymphocytes most IKKα is phosphorylated at the canonical residues that regulate proteasome-mediated degradation and destabilization, whereas in CD30lo lymphocytes most IKKα and IKKβ is unphosphorylated at these same residues (C).
NF-κB transactivates Meq transcription in vitro
Because we proposed a feed-forward loop model of increasing Meq and CD30 expression
Figure 5
QPCR based results of various transcription reporter assays and phylogenetic tree constructed from CD30 promoter sequences of MD resistant and susceptible genotypes
QPCR based results of various transcription reporter assays and phylogenetic tree constructed from CD30 promoter sequences of MD resistant and susceptible genotypes. QPCR results from in vitro transcription assay comparing the efficacy of NF-κB subunits and Meq combinations on transcription from the MDV RB-1B Meq promoter (mean ± sem); a-f indicate statistical differences (A). Unrooted phylogenetic tree constructed from CD30 promoter sequences for MD-resistant (61, N) and MD-susceptible genotypes (7, 72, 15I5 and P) (B). Quantitative effects of Meq on transcription from the MD resistant and susceptible genotypes' CD30 promoter (mean, 95% CI) (C).
Meq-dependent differential CD30 promoter transcription
It would be reasonable that differences in the CD30 promoter could confer differences in Meq-induced activation or repression of the CD30 gene and is of interest to us because of chicken genotype differences to MD lymphomagenesis after MDV infection. To measure Meq-induced CD30 transcription on different CD30 promoters, we first cloned and sequenced CD30 promoters (2431–2438 bp 5' to the ATG) from two MD-resistant (lines 61 and N) and four MD-susceptible (line 71, 72, 15I5, and P) genotypes of chickens and sequenced these. An unrooted phylogenetic tree (Figure
After cloning into an expression plasmid, each CD30 promoter was used in in vitro transcription assays using a Meq-expressing plasmid. Meq altered transcription from all CD30 promoters' alleles (Figure
Meq interacts directly with proteins central to lymphomagenesis
Meq's functions are modulated by its interacting partners. Here we wanted to identify which proteins were involved with Meq in the context of DNA binding and so we used chromatin immunoprecipitaion (ChIP) using anti-Meq antibodies (rather than traditional immuneprecipitation), followed by 2D LC-MSMS. We used MSB-1 MDCC cells as a model for tumor cells. We identified 31 proteins (PRIDE Accession # 14847–14852, Additional file
Figure 6
Meq interactome showing previously published and novel interactions identified by ChIP experiments and GO based modeling of Meq interacting proteins
Meq interactome showing previously published and novel interactions identified by ChIP experiments and GO based modeling of Meq interacting proteins. Meq interactome based on combined data from proteins identified by chromatin immunoprecipitation (ChIP) experiments (see Methods) and published literature. Differential expression is shown for proteins identified by ChIP. (A). Meq interactome converted to a GO functional network showing the physiological processes that Meq directly affects are consistent with neoplasia. Circle size represents effect size based on numbers of proteins associated with that GO annotation (B).
Evidence supporting a direct mechanistic connection between inflammation and cancer has been mounting over the last decade
In vitro MDV Meq induced CD30 expression and persistently activated NF-κB and ex vivo-derived CD30hi lymphocytes have increased and activated NF-κB protein. Not only does Meq enhance its own transcription (as previously described
Regardless, our data supports our hypothesized model that Meq initiates a self-reinforcing CD30 signaling cycle resulting in constitutive and aberrant NF-κB activation and subsequent neoplastic transformation. Herpesviruses co-evolve with their hosts and and the last common ancestor between EBV and MDV was at least ~300 M years ago; MDV, EBV and KSHV have separately evolved in different target cells the same fundamental result by targeting the NF-κB pathway. Furthermore both MDV Meq and EBV LMP-1
Meq is essential for MD lymphomagenesis
Non-transformed cells are common in lymphomas of all species, and often they form the majority cells in lymphomas. Our work suggests that many of these non-transformed cells are likely not immune responding cells in MD
Overall, and in the context of understanding mechanistic details of CD30hi lymphomagenesis, our results provide direct ex vivo-derived support in a natural animal model for the in vitro results in other species, which propose that ligand-independent and -dependent CD30 signaling-induced constitutive activation of NF-κB is a mechanism of neoplastic transformation in Hodgkin's disease antigen over-expressing lymphomas
Here we identify the neoplastic and non-neoplastic component of lymphoma microenvironment using transcriptomics and proteomics followed by Systems Biology modeling to generates specific hypotheses and then tests these using reductionist methods. This work provides evidence that MD neoplastic transformation is a continuum and the CD30lo lymphoma cells are in various stages of neoplastic transformation towards CD30hi phenotype. We hypothesized that MDV uses its Meq oncogene to activate CD30 transcription to achieve constituent NF-κB signaling resulting in cellular instability and a neoplastic phenotype. Our results show that Meq, CD30 and NF-κB proteins are overexpressed in CD30hi cells and that the majority of NF-κB is intranuclear suggesting an activated state. Using transcription reporter assays, we further show that NF-κB isoforms differentially activate Meq transcription, and Meq and NF-κB isoforms have additive effects. We also show that Meq transcriptionally activates or represses the CD30 promoter depending upon the host genotype from which the promoter is derived. Using ChIP and mass spectrometry we propose a new Meq interactome composed of proteins which are involved in various biological processes inherent in neoplasia. Overall, this study provides crucial insights into various molecular mechanisms of neoplastic transformation active within a heterogeneous lymphoma microenvironment in a natural animal model with functional immune system.
RNA isolation and microarray experiments
Lymphomas were isolated from white leghorn chickens infected with MDV GA/22 strain as described
Protein isolation and protein analysis by 2 dimensional liquid chromatography electro-spray ionization tandem mass spectrometry (2D LC ESI MS/MS)
Proteins were isolated from three replicates from (from different birds) 107 CD30hi and CD30lo cells using differential detergent fractionation (DDF), trypsin-digested and analyzed by 2D LC ESI MS/MS using a LCQ Deca XP Plus (Thermo Fisher Scientific) as described
Co-immmunoprecipitation of Meq interacting proteins
Meq interacting proteins were identified by chromatin immunoprecipitation (ChIP) assays with polyclonal anti-Meq antibody. MSB-1 cells were grown in Leibowitz's L-15 and McCoy 5A media supplemented with fetal bovine serum (15%), penicillin (100 IU/mL) at 37°C. Cells (107) were cross-linked with formaldehyde (1%, 10 min, 37°C), which was added directly to the culture medium. The culture medium was removed and washed twice with ice cold phosphate-buffer saline (PBS) containing protease inhibitor cocktail (Sigma-Aldrich, P8340). ChIP was done using the Chromatin Immunoprecipitation Assay kit (Millipore, 17–295) exactly following manufacturer's recommendations. Immunoprecipitation was performed with anti-Meq polyclonal antibody (1:100), incubated overnight at 4 °C. The DNA/Meq/antibody complexes were purified using Protein A-agarose/salmon sperm DNA beads. The purified complex sample was reverse cross-linked separating the DNA from Meq and its interacting proteins. Proteins that were co-immunoprecipitated with Meq were analyzed and identified by 2D LC ESI MS/MS as described above.
Plasmid construction
The CD30 promoters (2431–2438 bp upstream from the ATG start codon) of six different chicken lines (61, 71, 72, 15I5, N and P) were amplified by PCR with Pfu polymerase (Promega) and primers CD30-F and CD30-R. The amplified promoters were ligated into pCR®2.1-TOPO® (Life Technologies) producing pCR®2.1-CD30 plasmids. The cytomegalovirus (CMV) promoter in the pd2EGFP-N1 plasmid (Takara Bio) was removed by digestion with XhoI and VspI; linear DNA was blunt-ended by T4 DNA polymerase and then self-ligated producing pd2EGFPΔCMV. CD30 promoters were released from the pCR®2.1-CD30 plasmids by EcoRI digestion and ligated into EcoRI-linearized pd2EGFPΔCMV resulting in production of the six new expression plasmids pd2EGFP-CD30. The Meq promoter (1 kb upstream from the Meq start) of the virulent MDV-1 strain RB-1B was amplified by PCR with primers MEQ-F and MEQ-R. The promoter was first cloned into pCR®2.1-TOPO®, then released by EcoRI digestion and re-cloned into EcoRI-linearized pd2EGFPΔCMV producing the reporter plasmid pd2EGFP-Meq. The chicken cDNA encoding the NF-κB p100
Promoter assays
The activity of CD30 and Meq promoters was analyzed in vitro by promoter reporter assays. First, the reporter gene d2EGFP was placed under the control of the CD30 and Meq promoters and the coding sequences of transcription factors (Meq oncogene, and NF-κB p65, p100 and p105) were cloned into the expression plasmid pBK-CMV (see Construction of plasmids above). The promoter reporter plasmids and transcription factor expression plasmids were then transfected into SOgE cells
SOgE cells were grown in Dulbecco's modified Eagle's minimum essential medium supplemented with 10% fetal calf serum, penicillin (100 IU/mL), streptomycin (100 mg/mL) and amphotericin B (0.25 μg/mL) at 37°C with 5% CO2. Plasmids were transfected in triplicate into SOgE cells in 24 well plates at 80% confluence using Lipofectin® reagent (Life Technologies) following the manufacturer's instructions. Each well was transfected with 200–400 ng of DNA. To determine the effect of the Meq oncogene on the activity of the chicken CD30 promoters SOgE cells were transfected with either pUC18 alone (200 ng/well, negative control), pd2EGFP-N1 alone (200 ng/well, positive control), pd2EGFP-CD30 alone (200 ng/well, to measure the baseline level of expression from each of the CD30 promoters), or with a mix of pBK-CMV-Meq and pd2EGFP-CD30 (200 ng of each plasmid/well). To determine the transactivation effect of the NF-κB transcription factors alone or in combination with the Meq oncoprotein on the Meq promoter SOgE cells were transfected with plasmid mixtures and DNA. Plasmid pUC18 was added to transfection mixtures to give total amount of 400 ng plasmid DNA per well whenever it was necessary. Total RNA was isolated from transfected SOgE cells 48 h post transfection using TRI reagent (Molecular Research Center) following the manufacturer's instructions. Isolated RNA was treated with DNaseI, extracted with phenol/chloroform, precipitated with ethanol and resuspended in water.
The d2EGFP mRNA levels in transfected SOgE cells were quantified using the Platinum Quantitative RT-PCR ThermoScript One-Step System (Life Technologies). Both, d2EGFP and 28S rRNA amplicons, were designed using Beacon Designer (PREMIER Biosoft). The reaction mixture consisted of 2X ThermoScript Reaction buffer, 10 μM of each primer, 1 μM each of probes, Platinum
Gene ontology (GO) based phenotype modeling
GO was used to identify the phenotype of CD30hi and CD30lo cells, specifically with respect to GO terms which are associated with cancer
mRNA and protein expression comparison
We calculated the fold change in amount of mRNAs and proteins transcripts in CD30hi cells compared to CD30lo cells in semi-quantitative manner. For microarray data we calculated the fold change in terms of ratio of normalized fluorescent intensities; for proteomics data, fold change was calculated by taking the ratio of mean sum of XCorr of that protein in CD30hi to CD30lo cells.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
JB did chicken infection experiment, RNA and protein isolation. SK and JB did protein isolation. SK did microarray, proteomics, and systems biology data analysis and wrote the manuscript. DK did plasmid construction, and transcription reporter assays and SNP assays. HI did the microarray experiments and data analysis. KP did the proteomics 2D LC ESI MS/MS experiment and data analysis. SS did ChIP assay. SB conceived the project, did data analysis and wrote the manuscript. HC and HZ provided the necessary support for microarray, western blot and ChIP experiments and reviewed the manuscript. All authors read and approved the final manuscript.
This paper was supported by USDA NRI 2006-35204-16549.
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Oil Pan Too Weak
CarComplaints.com Notes: Although it has a stellar record in terms of vehicle problems, initially the 4th generation (2003 - 2009) 4Runner didn't earn our top "Seal of Awesome" rating because it received poor crash test ratings for rear impacts. However it's proven to have one of the lowest driver death rates out of all vehicles (not just SUVs), so we've bumped it back up to our top award.
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Herbert Walwick is a character in L.A. Noire. He is a patrolman.He is seen arresting Cliff Harrison in the A Slip of the Tongue case.


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This is a list of bosses in New Super Mario Bros. Koopaling Chaos.
|Pom Pom
|Pom Pom is one of Bowser's minions, and Boom Boom's partner.
|World 1 Tower
|Wendy O. Koopa
|Wendy gets everything she wants- except defeating Mario.
|World 1 Castle
|Negative Wendy O. Koopa
|Negative Wendy is a more powerful version of Wendy.
|World 1 Secret Castle
|Boom Boom
|Boom Boom is Pom Pom's long time partner.
|World 2 Tower
|Ludwig von Koopa
|Ludwig is odd and "kooky".
|World 2 Dragon's Den Castle
|Negative Ludwig von Koopa
|Negative Ludwig is even more kooky, and you could even say creepy.
|World 2 Secret Tower
|Boom Boom and Pom Pom
|The two partners working together!
|World 3 Tower
|The Robot Operating Buddy is mad at Mario... For some unknown reason. (Maybe He Wants Revenge From Deafating Him In SSBB)
|World 3 Factory
|Iggy Koopa
|Iggy has been obsessed with magic ever since Bowser got him a wand!
|World 3 Pyramid Castle
|Koopa Kid
|Koopa Kid wants revenge on Mario from the Mario Parties!
|World 3 Secret Factory
|A thunder Lakitu.
|World 4 Tower
|Mecha Bowser
|A new version: 2.0! The last one mysteriously disappeared...
|World 4 Factory
|Morton Koopa Jr.
|The first of the Jrs, oddly seen wanting lots of gold in this game...
|World 4 Gold Castle
|Koopa Kid 1, 2, and 3
|Koopa Kid can split up into three!
|World 4 Airship
|Negative Morton Koopa Jr.
|The first of the Negative Jrs, who likes gold too... If it is flaming!
|World 4 Secret Airship
|Lemmy Koopa
|Lemmy likes magic just a little less than Iggy.
|World 5 Tower
|Wario is Mario's evil twin.
|World 5 WarioWare Factory
|Ultimate Lemmy Koopa
|Still the same person, but ultimately "undefeated"!
|World 5 Circus Castle
|Morton Koopa Sr.
|Who do YOU think Morton Jr. was named after?
|World 5 Hidden WarioWare Factory
|Waluigi is Luigi's evil twin. If Mario has one, Luigi does too!
|World 6 Tower
|Pink Boo
|A boo that is pink.
|World 6 Haunted Tower
|King Boo
|The king of all the boos!
World 6 Luigi's Castle
|Dry Morton Koopa Sr.
|You defeated him. Now he's been skeletonized.
|World 6 Hidden Haunted Tower
|Wario Brothers
|The evil brothers want REVENGE! Escape from their monster truck... If you dare!
|World 7 WarioWare Factory
|Bad. Yoshi. That's how he got his name.
|World 7 Island
|Cosmic Clone
|He can gain your power-ups too!
|World 7 Tower
|Roy Koopa
|The strongest, meanest, KOOPALING... Oh wait, I dropped my papers...
|World 7 Arena Castle
|Uh, still got to get this in order... Okay. Magic Vs. Mario = Uh-oh!
|World 8 Tower-Mushroom
|Larry Koopa
|Oh, so sad... The LAST of the Koopalings... Probably, I haven't found all my papers yet...
|World 8 Castle-Mushroom
|Head of the Koopatrol
|World 8 Tower-Flower
|Bowser Jr. In Koopa Clown Car
|Who could forget Bowser Jr, the second Jr? I did.
|World 8 Castle-Flower
|Shadow Mario
|He was Bowser Jr. Who is he now?
|World 8 Tower-Leaf
|Metal Mario
|Not a power-up anymore.
|World 8 Castle-Leaf
|All of them?!
|World 8 Castle-Koopaling
|Negative Koopalings
|Defeat all of the Negative Koopalings separate and you can get to them!
|World Nega Tower
|He wants you... To be defeated!
|World Nega Castle
|Dry Bowser
|The end... Of you!
|World Nega Bowser's Castle


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Modfire vs. Target
Written by Katie on March 19, 2014. Permalink
Ouch. It physically pains me to see blatant rip-offs*. On the left, you have Modfire's Urbanfire; on the right, Target's recent Threshold™ Chiminea. Notice that trademark symbol? Not cool. For more information, visit Modfire's facebook page.
Purchase Information:Price: $1,350.00
Available from: Modfire


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6547 Gran Via Drive NE, Rockford MI, 49341
Sold Price:
3 beds , 2 Full/1 Half baths
3,100 sqft
Status: Closed
Last Updated: 7/27/2017 1:57 PM
Description for 6547 Gran Via Drive NE Rockford, MI 49341
Located in the desirable Lake Bella Vista community, this home has 3 bedrooms, 2.5 bathrooms, and over 3,000 square feet of living space. It is located less than a block away from the common area with beach/lake access to all-sports Lake Bella Vista. Open floor plan on the main floor has spacious family room with wood-burning fireplace, formal dining room, kitchen, half bath, and flex room perfect for an office, playroom, or formal living room. Hardwood floors run throughout the foyer, kitchen, and dining room. Upstairs are 3 bedrooms and 2 full bathrooms including the master suite with large walk-in closet, jetted bath, and separate shower. Basement is finished and has an open living space suitable for a theater room/recreation area and a 4th non-conforming bedroom.
Sold Price:
Sold Date:08/14/2015
Cannon Twp
2 Full/1 Half
3,100 (approx)
Listing Information
Cannon Twp
Other, Public Sewer
Cable TV, Natural Gas
School Information
Room Information
Family Room
Eat-In Kitchen, Butler Pantry
Interior Features
Gas Range/Cooktop, Dishwasher, Dryer, Convection Oven, Range, Refrigerator, Washer
Central Air, Ceiling Fan(s)
Gas Heat, Forced Air
Basement, Full
1, Wood-burning
Wood Floors
Exterior / Lot Features
Attached Garage, 2 Car Garage
In Ground Sprinklers
115 x 113
Lake, Garden, Beach Front, Corner, Paved Road, Acreage
Community Features
Park Play Area
Driving Directions
North on Meyers Lake Drive, West to Gran Via Drive, home on corner of Gran Via Drive and Laguna Vista Drive.
Financial Considerations
Listing Price History
Rockford Real Estate
Rockford is located in Michigan. Rockford, Michigan 49341 has a population of 6,134.
The median household income in Rockford, Michigan 49341 is $48,641. The median household income for the surrounding county is $52,716 compared to the national median of $53,482. The median age of people living in Rockford 49341 is 32.6 years.
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Помогите нам: эти порнозвезды присутствуют в этом видео?
Pornstar Name:
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she is moaning before she is even getting fucked L0L!@!@!@!@!@!
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this chick faked it just to get him off haha
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This means if your child has a fever at 3pm or 3am they should NOT come to school the next day. This is to prevent the spread of illnesses that are contagious during a fever. We love our students and love that they want to be at school, but this is to prevent spread of illnesses to others.
Also, it is not appropriate to send ANY over the counter medicine with a student to take at school unless it has a doctor's order/note and parent signature and nurse has checked them.
Thanks for helping us stay healthy together!
Carson James, a 6th grade band student, performed The Star Spangled Banner on his alto saxophone before the basketball game versus Porter Ridge Middle School
On November 29, 2016, EUMS held their annual School Spelling Bee. Students from each grade level were represented. Several family members attended and watched the students compete. After over an hour of competition, and nearly 200 words, we were able to name a winner.
The East Union Robotics Team and Mrs. Davis's classes competed in the 2016 Fall STEM Competition on Friday, November 18 at SPCC. Events included Dragster, Catapult, Bridge Building, and Robotics.