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Our Michigan medical malpractice lawyers represent clients who have suffered injuries, such as blindness, due to a doctor's carelessness when controlling and maintaining a patient's diabetes. Not properly controlling a patient's diabetes is very serious, often requiring extensive medical treatment. When a medical error causes a death, it can give rise to a Michigan wrongful death lawsuit.
Poorly controlled or uncontrolled diabetes often results in blindness of the patient. Blindness resulting from poorly controlled diabetes is referred to as diabetic retinopathy. According to the CDC, diabetes is the number one cause of adult blindness in the U.S. resulting in 24,000 new cases each year.
What are the causes of Diabetic Retinopathy?
People with all types of diabetes (type 1, type 2, and gestational) are at risk for diabetic retinopathy. The risk of the condition increases the longer a person has diabetes. Studies estimate that up to 45 percent of Americans diagnosed with diabetes have some stage of diabetic retinopathy. Many people are unaware they even have the condition.
The high levels of blood sugar weaken tiny blood vessels in the eye, eventually causing them to bleed. This process leads to diabetic retinopathy. Women who develop or have diabetes during pregnancy may have rapid onset or worsening of diabetic retinopathy.
This bleeding damages the retina, the part of the eye that receives light and sends images to the brain, and can distort vision. In severe cases, it can lead to permanent blindness and loss of vision. Many times, the condition is permanent.
What are the Types of Diabetic Retinopathy Lawsuits?
A person with diabetic retinopathy can file a lawsuit against a physician, optometrist, or ophthalmologist for failing to properly diagnose and treat the condition at an early stage in the disease process. The condition can be detected during a comprehensive dilated eye exam. A trained doctor should be able to diagnose the condition and initiate early treatment to prevent permanent harm to the eyes. The failure to do this may constitute medical negligence and give rise to a medical malpractice lawsuit.
To win this type of lawsuit, it must be proven that the medical provider was negligent in failing to diagnose diabetic retinopathy or in failing to refer the patient to a specialist with the expertise to make an accurate diagnosis. It must also be proven that an earlier diagnose would have led to treatment that would have minimized the severity of the harm to the eyes and vision. We find the top medical specialists in the United States to review the medical records and give expert opinions on these issues.
How much are Diabetic Retinopathy Lawsuit Settlements?
There is no formula or settlement calculator to determine the proper compensation in your case. Many factors are used to determine the amount of an eye injury settlement, including:
- The type of negligence that caused the condition
- The permanency of the condition
- The severity of the harm (complete vs. partial blindness)
- How the condition has affected the victim's life (inability drive, work, read, etc.)
- The age and occupation of the patient
- Whether the person has suffered a loss of income or earnings, and if so, how much money?
- The amount of medical expenses (past, present, and future)
- The type and amount of insurance coverage available to pay your settlement.
Eye injury cases often result in substantial settlements due to the severe impact on a person's life. Our attorneys will discuss your case with you, evaluate the factors of your case, and give your our thoughts and opinions about the settlement value of your case.
Best Michigan Diabetic Retinopathy Lawyers
Our legal team includes the most successful medical malpractice attorneys in Michigan. Buckfire & Buckfire, P.C. and our lawyers have won the top awards in the legal profession, including:
- U.S. News & World Report Best Law Firms
- "Best Michigan Personal Injury Attorney" by American Lawyer Academy
- Named Top 100 Trial Lawyers in Michigan
- Honored as "Super Lawyers"
- Top Martindale-Hubbell Rating (AV) for Skill & Integrity
Our Michigan Lawyer No Fee Promise to You
Our Michigan eye injury attorneys will represent you in your case under our NO FEE PROMISE. This means that you will not be charged any legal fees whatsoever unless we obtain a settlement or recovery for you. You have absolutely no obligation for legal fees unless we win your case. If we do not obtain a settlement for you, you owe us nothing!
Contact Your Michigan Diabetic Retinopathy Lawyers
Our Michigan lawyers are experienced trial attorneys who know what it takes to recover the maximum amount of compensation for your case. If you or a loved one has suffered blindness or loss of vision due to undiagnosed or untreated diabetic retinopathy, call us at (888) 797-8787 to learn about your legal right to a settlement.


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Monday October 21, 2013 CrossFit Stapleton- Denver, CO
Happy Birthday Ann L and Leslie G! We hope you both have a great day!
– CFS Staff
Tay's Paleo Pumpkin Spice Latte Creamer
8oz Canned Coconut Milk (Full Fat)
8 dates (pitted)
1 teaspoon chia seeds
1-2 teaspoons honey
1/2 cup canned Pumpkin
1/2 teaspoon vanilla extract
1 teaspoon cinnamon
1/2 teaspoon ginger
1/2 teaspoon cloves
1/2 teaspoon nutmeg
Blend all ingredients together in food processor. Add 1 tablespoon (or more!) to your 8oz cup of hot coffee. Whisk in coffee until fully incorporated. Enjoy!
Store extra creamer in an airtight container for up to 7 days in the refrigerator.
A) Squat Clean 3-2-2-1; start @75% 1 RM and build; Rest 3 B/T
B) EMOM Box Front squat 20X1 x 2-3 60-70% of 1RM x 6 mins
8 Rounds For Time:
8 Clapping Push Ups
10 Med Ball Cleans 20#/14#
8 T2B OR 12 Sit Ups not anchored

subset: commoncrawl


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As part of a requested restructuring, the DNC would like to introduce our new representatives at the DTES LAPP table, Co-Chairs Richard Cunningham and Jacek Lorek.
Richard Cunningham is President of the DNC, celebrated community advocate, DTES Street Market leader, long time DTES resident and champion of the low income community.
Jacek Lorek is Sargeant-At-Arms of the DNC, coordinator of the DTES Street Market, long time labor organizer, homeless shelter rights advocate, and Gastown resident.


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On selected Saturdays and Sunday during the year, an Ackland Student Guide leads an "Encounter Art" tour, helping visitors gain a deeper understanding of selected works in the Museum's permanent collection based on a particular theme.
Meet in the Ackland lobby. Free and open to the public.
Saturday, 16 April 2016:
- 3:00: "Beauty and the Beast," designed and led Ashley Wee
- 3:30: "Some Assembly Required," designed and led by Anthony Bowman
Sunday, 24 April 2016:
- 3:00: "The Art of Dance," designed and led by Lea Efird
- 3:30: "Ancient Selfie," designed and led by Jasper Goh


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Elsie is TWO! (and a Birthday Park Playdate)
May 29, 2013
Elsie Grace,
Why hello there sweet little girl?! I truly cannot believe that it was TWO YEARS ago that you came into this world and forever changed our lives.
Elsie, you truly have been adding sweetness to our days since you're birth. Our prayer is that you continue to do this very thing every day of your life. {Because everyone could use a little more sweetness in their life?!}
This past year has been full of kisses, dirty diapers, smiles, cheese (yes... cheese is your favorite food), dress up, tantrums and tears, memories, play dates, time impressing your future boyfriend/hubby, Jackson, wars and hugs with your sister, dance parties and most of all, incredible amounts of JOY and GRACE.We absolutely love watching you grow and standing on the sidelines as God works His way in you. Your spirit is oh so very sweet and we're convinced that you'll be the one daughter who won't ever move too far away from her mommy. :) (Can we guarantee that now?!)
For your birthday this year, we spent your actual birthday celebrating with our little family. You opened your gifts (with lots of help from your big sister) and indulged in a red velvet cupcake. The following day we spent the morning celebrating with friends at a nearby park where we played and enjoyed a little birthday cake (all while wearing a pink tutu, because every birthday girl gets to wear a tutu to the park on her special day!).
All in all, it was a great celebration of your life!
Just as we know that God delights in you, we too delight in you day in and day out.
mommy and daddy


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What is needed of a tour host? It's really quite simple. Our coordinators for that particular genre will send out an email saying that they are scheduling a tour for a book and will give you the info. After you are signed up, we send you all the info you will need (Except for the post from the author. That will come closer to your stop date.) You also get the chance to review the book for the tour so we would send you the book in advance. On the date of your stop, all you have to do is post the items that the coordinator has sent. It's as simple as that. There is usually the opportunity to give away a copy of the book that is being toured. (Unless specified otherwise.)
It's a great opportunity to be able to help the author promote their hard work and dedication they have put in the form of words. So if you are interested, please fill out the form below. We are currently looking for a few bloggers to sign on for a couple of tours. Check out our BTS Tour Host site for more info on the tours.


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I'm a practitioner in the Usui System of Natural Healing in accordance with the Usui Shiki Ryoko tradition.
Reiki is loosely translated to "universal life energy".
Usui Shiki Ryoko is a specific form of practice that allows the student to have contact with the energy of Reiki simply by laying on of hands in self-treatment and treatment of others. Most Reiki treatment involves the practitioner's hands being held on or near a specific part of the body for a varying length of time.
Contact me with any questions you have about the Reiki process or to set up an appointment.
Click here for a more information on the history of Reiki


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Grounds Maintenance
Championship Enterprises Inc. provides quality design services for residential, commercial and municipal properties. Our experienced and knowledgeable staff will work with you to ensure all of your facilities management services are met.
Call us for a consultation to discuss your unique commercial or residential needs. Our services include planting design, landscape and hardscape design and irrigation design. Our experienced staff are with you from start to finish.
We will:
- Conduct on-site measurements of the area or areas that you would like to work with
- Perform a site analysis
- Document all of the existing conditions with pictures and notes
- Schedule a follow-up visit at your home or office
- Review and revise your plan, provide a cost proposal and scale the project to balance your design and budget goals.


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Nina family release statements
Nina's family has reacted to the emotional meltdown she suffered yesterday.
It may seem as though her romance with Miracle is fast deteriorating.
Nina's family has appealed to her thousands of fans never to give up on her as she ends her romance with her fellow housemate, Miracle.
Nina revealed yesterday night to Tobi that she is done with Miracle.
Miracle has often times questioned the loyalty of Nina to him as she maintains her allegiance to her original lover, Collins.
Meanwhile, Miracle is ready to continue the relationship outside the house.
Read the two separate public statements made by her family via her Instagram account below:
Hi Ninafans, Ninalovers, ninanation and all our supporters! We are sure everyone has been overwhelmed with the drama going on in the house! It's been intense but let's all realise that even blood SIBLINGS have their own misunderstanding and no matter how long, they always find their way back if theres real love & trust. Let's not be too quick to judge or tarnish any image all in the name of a show. Let's all strive to be positive both in our comments and actions.
There's always going to be disagreements but the beauty of it all is we will all agree eventually! The parties involved are making decisions right now solely on emotions and the best we can do is support them, criticise positively and keep loving them.
It's two weeks to the finale! Let's not use the judgement of one day to jeopardize this three months dream we have all built. Let's drop the negativity, show love, use positive words and keep Nina's dream alive by supporting her always.
Nina is a human being and not incapable of learning; whatsoever it is that she isn't doing well, when she comes out, she will learn. Had she done or said things that made you angry as fans, it's time for you to overlook those imperfections… Are those things enough to make you not to vote for Nina next week? As Nina fans and Lovers, Please look beyond her imperfections and never give up on her.
Nina is a human being even admins are also human too . The emotional roller coaster are sometimes uncontrollable but isn't it why we love the show? When young people are living together, disagreements are inevitable; may God give Nina more grace to carry on to make us all happier. Good morning and have a blessed day…


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The Bouvier Family is one of the main families in Springfield.
Marge's parents are Clancy Bouvier and Jacqueline Bouvier (nee Gurney). It has been proven by Marge in the episode "The Color Yellow," when she told Homer, "You're not French, my Dad is." This means that Marge and the kids are of French descent.


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Ministry to Special Needs
Uganda has over 4 million children with known special needs (like; cerebral palsy, autism, epilepsy, HIV/AIDS, blind and many more). Orphans and children from very poor families are more at risk, many of such children are abandoned and some die in the early years of their childhood since most parents do not know how to handle such kids at the same time they don't have the financial resources to care for these kids and the government does not provide the necessary help to such parents.
We identify the most vulnerable children with special needs and we give them the required help ranging from medical care, education or place them in rehabilitation homes as need be.


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A Little Horse
We decided to bunk off school last week. We've been nose to the grindstone for a number of months and we thought it was about time we took a slight breather and celebrated a few recent successes.
So we all snuck off to the Albion in Islington and sat in the beer garden all afternoon. Drinking rose, eating great gastro BBQ pub food (although Pigs Ears as starters definitely didn't count and aren't to be recommended) and playing games.
Now as the drink flowed and as a new team we had a lot of fun playing "tell us something we don't know", which ended as "we've just learnt things we wish we didn't know".
Obviously discretion is the better part of valour (and we wouldn't want all our Clients resigning), but there were lots that involved the police, the opposite sex, the same sex, no sex, poo, pooh, phew and even a date with Robbie!
We then had a brisk walk round to Little Voice, where we proceeded to act out the final of Britain's Got (No) Talent. After being informed that we were going to for a sing-song, the some of the team were somewhat disheartened and not really that interested. It's amazing what a few bottles of rose does to ones potential stage fright.
Some 3 hours later we finally gave back our microphones, blow up guitars, wigs and hats, but we hung on to our slightly hazy bizarre memories, including Strings MC'ing a duet with Hamish, Claire wrestling on the floor as she refused to hand back her Mike, Nick being a Rock God, John being too Sexy for his shirt, and Mandy (Holden) actually getting lost in music.
Basically we sang until we all lost our voices, drank till we couldn't drink anymore and all thoroughly had a brilliant night out – Here's to celebrating success.


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FY 2018 Annual Report
PADD Hazard Mitigation Plan
Directions to PADD
KY Council of ADD's
West Kentucky Future
Hello and welcome here! This is the Purchase Area Development District's Info & Data Center.
Here you will find a variety of information about the PADD itself, our service region as a whole and for each of our 8 counties. Hopefully, you will find what you are looking for!
Here you will find information about the Purchase Area Development District (PADD).
Here you will find information and statistical information about the Jackson Purchase Area as a whole.
Here you will find information and statistical information about the individual counties in our region
Here you will find information and links about tourism opportunities and Points of Interest in the Purchase Area, along with an interactive map, event listings and much more.
Click here ...
Here you will find other useful materials and information.
Graves County
Graves County, 75th in order of formation, is located in the center of the Jackson Purchase on the Tennessee state line and is the largest county in that region. It is bordered
by Hickman, Carlisle, Marshall, McCracken and Calloway counties and has an area of 555 square miles. Established in 1824, it was named in honor of Major Benjamin F. Graves
of Virginia, who was a state legislator and a Fayette county soldier killed at the Battler of River Raisin in the War of 1812. Its county seat is Mayfield, Kentucky.
Other Information:
Population (2010): 37,121
Land Area: 555.59 Square Miles
Water Area: 0.87 Square Miles
Population Density: 66.81 people per square mile
Cost of Living Index: 81.2 (low, U.S. average is 100)
Judge Executive Office
Judge Executive
Jesse Perry
101 East South Street
Mayfield, KY 42066
Phone: (270) 247-3626
Fax: (270) 247-7653
Email: email@example.com
Website: http://gravescounty.ky.gov
Office Hours
8:00 AM - 4:30 PM Central Time
Fiscal Court Meetings
2nd + 4th Monday at 4:30 PM
County Clerk:
City of Mayfield
Kathy O'Nan
211 East Broadway
Phone Number:
(270) 251-6251
Fax Number:
(270) 247-2485
City Council Meetings:
2nd Monday at 6:00 PM
City of Wingo
Charles Shelby
230 St. Paul Street
Wingo, KY 42088
(270) 376-2286
3rd Monday at 6:30 PM
External Links for more Statistical Information
Graves County Census QuickFacts
Think Kentucky Demographics County Profile
Graves County City Data
United States Bankruptcy Court - Western District of Kentucky
Page last updated on Mon, 13 May 2019 13:36:00 GMT.


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The book 8 patch notes are finally live. You can read them all here. Below are the things that stick out in my mind.
Class balance changes: Runekeepers, and Minstrels to a lesser extent, are getting a damage nerf. Blackarrow DoTs will now be curable. Champions will get healed for less when in their highest damage stance.
Thoughts: The first two changes were needed. No-one should be able to out DPS a hunter on single targets, that is their primary class role. And Blackarrows apparently have been insanely overpowered since the last patch.
The Champion change is puzzling. No sane champion would use their high damage stance when trying to tank. All of your active defenses go to zero, and high AC is not nearly enough to make up for that in my experience. Even as an offtank, if you are in a situation where you are expecting to need frequent heals...again you aren't going to use your highest damage stance. You'll go for the middling one or for full defenses. This change ultimately makes little difference to the class, and I am not sure why Turbine thought it was necessary.
Lothlorian gift box changes: low end rewards have been removed, you will be guaranteed at least a rune worth 250K weapon XP. The possibility of first age weapons has also been removed. Instead, the chances of getting a second age have been improved. However, second ages that you get will randomly be 58, 59, or 60. The cost for gift boxes has also been reduce by nearly 40%.
Thoughts: The odds of getting a first or second age weapon out the boxes is so low that there is little reason to even try it in the current game. The low end rewards that you can get are also so bad that they might as well be coal. This sounds like a good change. However, until this goes live it remains to be seen whether the entire Lothlorian gift box system is worth messing with. If the chances of getting a 2nd age legendary are still only 5% or so, I'd say it's still not worth it.
Crafting Revamp: all optional items will now tell you what crafts use them in the description. Optional items will only drop off of humanoids and crafting nodes. Teir VI crafted armor is getting stat boosts.
Thoughts: Being able to tell what a crit item is used for by reading the description is absolutely a needed change. Most of these items currently only say "this item can be used by crafters." I have high level crafters in most of the professions, and half of the time I haven't got the foggiest idea which character to send a given crit item to. I absolutely applaud this change.
The change in drop patterns of crit items honestly bothers me. One of the things I enjoy about the current game is that if you want certain crit items, you have to head out and farm specific mobs. I think it's great that crit items are now available from resource nodes. However, I don't think it's so great that humanoids are the only mobs that will drop crit items now, and that they will drop all of them. It seems like some of the depth of the game has been diminished.
Finally, the stat boost on crafted tier VI armor. As you might surmise if you have read any of my past posts, I feel that this is absolutely a needed change. The current tier VI armor is actually so bad that if you have the best crafted tier V armor there is no reason to upgrade. Even the best guild tier VI armor is barely better than the best tier V armor. As it stands currently, a maxed out tailor can make a few good cloaks and a maxed out metalsmith can mainly make tools. Other than that neither profession makes any gear at all that most characters would care about. I am very glad to see Turbine addressing this. If the new recipes are on par with the normal mode coin drop gear, I'll be happy as a clam.
Now if only Turbine would revisit the PvP gear, we might have an end game as good as what we had 6 months ago.
Thursday, May 21, 2009
Book 8
Labels: Book 8, Lord of the Rings Online
2 comments:
I'm pretty excited about the crafting revamp. I actually like the drop changes they're planning, that way I don't have to go to the web to find out what creature drops the one optional ingredient I need.ReplyDelete
I don't play LOTRO anymore, but I like the idea of the items telling you what they are for. When I tried LOTRO briefly, my bags got filled rather quickly with all sorts of things that I had no idea what to do with. I just vendored most of them because it's just too much hassle, and no fun, to sit there looking each item online to see if what I can do with it. I'd end up spending more time in a browser than in game playing!ReplyDelete
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- Research
- Open Access
The Gαh-PLCδ1 signaling axis drives metastatic progression in triple-negative breast cancer
© The Author(s). 2017
- Received: 10 March 2017
- Accepted: 25 May 2017
- Published: 2 June 2017
Distant metastasis of triple-negative breast cancer (TNBC) to other organs, e.g., the lungs, has been correlated with poor survival rates among breast cancer patients. Therefore, the identification of useful therapeutic targets to prevent metastasis or even inhibit tumor growth of TNBC is urgently needed. Gαh is a novel GTP-binding protein and known as an inactive form of calcium-dependent tissue transglutaminase. However, the functional consequences of transamidating and G-protein activities of tissue transglutaminase in promoting cancer metastasis are still controversial.
Kaplan-Meier analyses were performed to estimate the prognostic values of Gαh and PLCδ1 by utilizing public databases and performing immunohistochemical staining experiments. Cell-based invasion assays and in vivo lung colony-forming and orthotropic lung metastasis models were established to evaluate the effectiveness of interrupting the protein-protein interaction (PPI) between Gαh and PLCδ1 in inhibiting the invasive ability and metastatic potential of TNBC cells.
Here, we showed that the increased level of cytosolic, not extracellular, Gαh is a poor prognostic marker in breast cancer patients and correlates with the metastatic evolution of TNBC cells. Moreover, clinicopathological analyses revealed that the combined signature of high Gαh/PLCδ1 levels indicates worse prognosis in patients with breast cancer and correlates with lymph node metastasis of ER-negative breast cancer. Blocking the PPI of the Gαh/PLCδ1 complex by synthetically myristoylated PLCδ1 peptide corresponding to the Gαh-binding interface appeared to significantly suppress cellular invasiveness in vitro and inhibit lung metastatic colonies of TNBC cells in vivo.
This study establishes Gαh/PLCδ1 as a poor prognostic factor for patients with estrogen receptor-negative breast cancers, including TNBCs, and provides therapeutic value by targeting the PPI of the Gαh/PLCδ1 complex to combat the metastatic progression of TNBCs.
- Gαh
- Metastasis
- PLCδ1
- Protein-protein interaction
- Triple-negative breast cancer
Triple-negative breast cancers (TNBCs) are a diverse and heterogeneous group of tumors that lack estrogen and progesterone receptors and HER2 gene amplification. Because they do not express these receptors, hormonal (e.g., tamoxifen) and targeted (e.g., Herceptin) therapies are not effective in treating TNBCs. Moreover, the majority of TNBCs are highly malignant, and only a subgroup responds to conventional chemotherapy with favorable prognosis [1–3]. Therefore, drug resistance and metastatic progression are major clinical issues in the successful treatment of TNBCs. Although high-throughput molecular analyses, including sequencing, pathway analyses, and integrated analyses of alterations at the genomic and transcriptomic levels, have improved our understanding of the molecular alterations involved in TNBC development and progression, the proper use of this knowledge for the rational selection of therapy remains challenging.
Gαh is a novel GTP-binding protein that is an inactive form of calcium-dependent tissue transglutaminase; a multifunctional protein that is ubiquitously expressed in various tissues [4]. Gαh transmits signals from activated G protein-coupled receptors, e.g., α1-adrenoceptor [5] and follicle-stimulating hormone receptor [6], to phospholipase C-δ1 (PLC-δ1) via protein-protein interactions (PPIs). Activation of the Gαh/PLC-δ1 pathway has been shown to elevate the concentration of intracellular inositol 1,4,5-triphosphate (IP3) and calcium [7], both of which are important secondary messengers in signal transduction and promote cell survival, growth, and invasion [8, 9]. In contrast, an abnormal increase in intracellular calcium due to cellular damage or other stressors has been shown to activate the transglutaminase feature of Gαh activity to catalyze the crosslinking of cellular proteins and ultimately induce cell death [10]. Recently, Gαh overexpression has been observed in breast [11, 12], colon [13], and ovarian cancers [14]; non-small cell lung cancer [15]; and renal cell carcinoma [16] and has been shown to be associated with advanced disease stages and metastatic spread. Importantly, the loss of the GTP-binding, but not transamidating, activity of Gαh was shown to inhibit cancer metastasis in malignant tumor cells [17], implying the critical importance of Gαh/PLC-δ1-dependent signaling in cancer progression. These findings prompted us to evaluate the feasibility of therapeutic inhibition of the PPI of Gαh with PLC-δ1 in combating cancer metastasis.
Here, we find that Gαh expression is causally associated with the metastatic potential of TNBC cells in vitro and in vivo and is strongly correlated with poor distant metastasis-free survival probability in patients with breast cancer. Notably, loss of the G protein function, but not the transglutaminase activity, abolished the Gαh-induced metastatic progression of TNBC cells. Interestingly, blocking the PPI between Gαh and PLC-δ1 using a synthetic peptide blocker corresponding to the binding-interface of Gαh effectively inhibited the metastatic progression of TNBC cells in vitro and in vivo. These findings provide a potential new strategy to develop anti-cancer agents for overcoming cancer metastasis.
Cell lines and cell culture condition
TNBC cell lines MDA-MB-468, MDA-MB-231, and MDA-MB-436 were cultured in Leibovitz's (L-15) medium (Gibco Life Technologies, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and incubated at 37 °C with free gas exchange with atmospheric air. TNBC cell lines HCC1143, HCC1599, HCC70, and HCC38 were cultured in RPMI-1640 medium (Gibco Life Technologies) with 10% FBS and incubated at 37 °C with 5% CO2. HS578T and 293T cells were cultured in DMEM with 10% FBS and incubated at 37 °C with 5% CO2. All cell lines were obtained from American Type Culture Collection (ATCC). All cells were routinely authenticated on the basis of short tandem repeat (STR) analysis, morphologic, and growth characteristics and mycoplasma detection.
Public database
Raw data for pathological information and prognostic values of the Gαh gene were downloaded from the PrognoScan database (http://www.prognoscan.org/). Datasets with p value < 0.05 by Cox regression analysis were included in a meta-analysis for Gαh gene expression. Transcriptional profiling for Gαh was obtained from The Cancer Genome Atlas (TCGA) database and subjected to statistical analysis against differential levels of Gαh mRNA in the subgroups. Kaplan-Meier analysis and the differential analysis of Gαh gene expression were performed on the SurvExpress website.
In vitro invasion assay
For invasion assays, Boyden chambers (Neuro Probe, Gaithersburg, MD, USA) were used according to the manufacturer's protocol. Briefly, a polycarbonate membrane (Neuro Probe) was pre-coated with 10 μg of human fibronectin (Sigma-Aldrich, St Louis, MO, USA) on the lower side and Matrigel on the upper side. Cells (1.5 × 104) were seeded in the top chamber in 50 μl of low-serum medium (0.1% FBS) containing drugs or DMSO. After 16 h, stationary cells were removed from the top surface of the membrane, whereas the migrated cells on the bottom surface of the membrane were fixed in 100% methanol and stained with 10% Giemsa solution (Merck Biosciences, Mendota Heights, MN, USA) for 1 h. The invaded cells were counted under a light microscope (×400, ten random fields from each well). All experiments were performed in triplicate.
Western blot analysis
Proteins (100 μg) were boiled for 5 min in SDS sample buffer (62.5 mM Tris [pH 6.7], 1.25% SDS, 12.5% glycerol, and 2.5% β-mercaptoethanol) and separated on 10% SDS-PAGE gels. After transferring to PVDF membranes, the proteins were incubated with antibodies against Gαh (GeneTex, Hsin-Chu, Taiwan), PLC-δ1 (GeneTex), or GAPDH (AbFrontier, San Diego, CA, USA). The immunoreactive bands were visualized using an enhanced chemiluminescence system (GE Healthcare, Pittsburgh, PA, USA).
GTP-binding assay
Membrane proteins (100 μg) were diluted with 500 μl of 5-fold diluted buffer B (prepared as instructed by the manufacturer of the membrane extraction kit) containing 1 mM sodium orthovanadate, 1 mM dithiothreitol (DTT), and 4% protease inhibitor cocktail (Merck Biosciences). The membrane proteins were then absorbed into GTP-agarose (Sigma-Aldrich) overnight at 4 °C. The GTP-agarose-absorbed complexes/proteins were subsequently analyzed by SDS-PAGE/Western blotting using antibodies against PLC-δ1 or Gαh.
In situ tTG activity assay
A 50 μl aliquot of cell homogenate proteins (10 μg) in coating buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 5 mM EDTA, and 5 mM EGTA) was added to each well of a 96-well ELISA plate (Apogent, Portsmouth, NH, USA) and was incubated at 4 °C overnight. To block the coated wells, a 200 μl aliquot of 5% BSA, 0.01% Tween 20, 0.01% SDS in borate-buffered saline (BBS) was added. After an additional 2 h of incubation at 37 °C, the plate was rinsed once with 1% BSA and 0.01% Tween-20 in BBS. To each well, 100 μl of HRP-conjugated streptavidin (Southern Biotech, Birmingham, AL, USA) (1:1000) in 1% BSA and 0.01% Tween-20 in BBS was added, followed by incubation for another 1 h at room temperature (RT). After the wells were rinsed four times with 1% BSA and 0.01% Tween-20 in BBS, the specimen in each well was incubated with 200 μl of TMB substrate (Sigma-Aldrich) for 10–20 min at RT. The reaction was stopped with 50 μl of 3 N HCl, and the absorbance at 450 nm was measured using a microplate spectrophotometer.
Peptide synthesis
TIPWNSLKQGYRHVHLL, a peptide corresponding to the PLC-δ1 amino acid sequence from 720 to 736, was synthesized with or without myristoylation at the N-terminus and purified to approximately 95% (Mission Biotech, Taipei, Taiwan). Ten millimolar synthetic peptide was dissolved in 1 part of 0.1% trifluoroacetic acid in dd-H2O mixed with 1 part 0.1% trifluoroacetic acid in acetonitrile, aliquoted, and kept at −70 °C as a stock solution.
Plasmid construction
The genes encoding Gαh and PLC-δ1 were amplified from human cDNA (Invitrogen) using standard polymerase chain reaction (PCR) with paired primers (Additional file 1: Table S1) and sub-cloned into the pLAS3w/Ppuro, pLAS3w/Pbsd, or pIRES2-EGFP vector. The Gαh-containing pIRES2-EGFP plasmid was used as a DNA template for site-directed and deletion mutagenesis. PCR for site-directed mutagenesis was performed with paired primers (Additional file 1: Table S1) using a pfu polymerase kit (Stratagene, La Jolla, CA, USA) (for R580A) or primer extension method [18] (for W241A). The PCR products were treated with Dpn1 endonuclease (New England BioLabs, Hitchin, Hertfordshire, UK) to digest the methylated parental DNA template. For the construction of a C-terminal deletion mutant of Gαh (∆657-687), a Gαh-containing pIRES2-EGFP plasmid was used as the template to amplify the target sequence using paired primers (Additional file 1: Table S1). The two PCR products were subsequently digested with NheI and EcoRI and then ligated into pLAS3w/Pbsd. The identities of individual clones were verified via double-strand plasmid sequencing.
Preparation and infection of lentiviral particles
All lentiviral vectors, including pLAS3w/Ppuro and pLAS3w/Pbsd and derivatives of shRNA vector (obtained from the National RNAi Core Facility Platform in Taiwan), were transfected into the packaging cell line 293T along with pMD.G and pCMVΔR8.91 plasmids using a calcium phosphate transfection kit (Invitrogen). After incubation for 48 h, the viral supernatants were collected and transferred to target cells, and the infected cells were cultured in the presence of puromycin and blasticidin (Calbiochem, San Diego, CA, USA) at 5–10 μg/ml in order to select the stably transfected cells.
Animal experiments
NOD/SCID mice were obtained from the National Laboratory Animal Center in Taiwan and maintained in compliance with the institutional policy. All animal procedures were approved by the Institutional Animal Care and Use Committee at Taipei Medical University.
For the in vivo lung metastatic colonization assay, cells were implanted into mice through tail vein injection at a concentration of 1 × 106 cells/100 μl of PBS. The mice were sacrificed, and the lungs were dissected for histological analyses at the endpoint. The metastatic lung nodules were quantified after H&E staining using a dissecting microscope.
For orthotopic lung metastasis of breast cancer, GFP/luciferase-expressing MDA-MB-231 cells (5 × 105), which were established by virally transducing cells with an EF1 promoter-driven firefly luciferase vector and an IRES-driven EGFP vector, were suspended in 50 μL of PBS and then were subcutaneously inoculated into the abdominal fat pad of each mouse. In vivo tumor images were captured with an IVIS imaging system (Caliper Life Sciences, Alameda, CA, USA) to measure the signal intensity from the GFP/luciferase vector. The mice were humanely sacrificed at the end of the experiments, and lungs were obtained for histological analyses. The tumors at the primary sites were removed for weight measurement.
Clinical samples and ethics statement
The breast cancer tissues used in this study were from Wan Fang Hospital managed by Taipei Medical University (TMU). Patient information, including gender, age, and histopathological diagnoses, was collected. The surgical specimens had been fixed in formalin and embedded in paraffin before being archived. We used the archived specimens for IHC staining. Follow-up of patients was carried out for up to 60 months. A four-point staining intensity scoring system was devised to determine the relative expression of Gαh and PLC-δ1 in the cancer specimens; the staining intensity score ranged from 0 (no expression) to 3 (maximal expression). All of the IHC staining results were reviewed and scored independently by two pathologists. The study was carried out with the approval of the Institutional Review Boards and with permission from the ethics committees of the institutions involved (TMU-IRB 99049).
Immunohistochemical staining analysis
Paraffin-embedded tumor sections (3 μm thick) were heated and deparaffinized using xylene and then were rehydrated in a graded series of ethanol with a final wash in tap water. Antigen retrieval was performed with a target retrieval solution (DAKO, Woodbridge, VA, USA) in a decloaking chamber (Biocare Medical, Concord, CA, USA). Endogenous peroxidase activity was quenched by hydrogen peroxide. The sections were then incubated with anti-Gαh and anti-PLC-δ1 antibodies (GeneTex) at 4 °C overnight. The Vectastain ABC peroxidase system (Vector Laboratories, Burlingame, CA, USA) was used to detect the reaction products.
Statistical analysis
SPSS 17.0 software (Informer Technologies, Roseau, Dominica) was used to analyze statistical significance. Paired t test was utilized to compare Gαh gene expression in the cancer tissues and corresponding normal tissues. Pearson's test was performed to estimate the association between mRNA and protein expression of Gαh and PLC-δ1 by RNA-sequencing and IHC, respectively. Evaluation of survival probabilities were determined by Kaplan-Meier analysis and log-rank test. One-way ANOVA with Tukey's test was used to estimate the difference in lung colonies and tumor weights in the animal experiments. The Mann-Whitney U test was used to analyze the non-parametric data. p values < 0.05 in all analyses were considered statistically significant.
Gαh upregulation predicts a poor probability of distant metastasis-free survival in breast cancer patients
Next, we examined the transcriptional profiles of Gαh in clinical tissues from breast cancer patients using The Cancer Genome Atlas (TCGA) database [20]. The data showed that the transcriptional activity of Gαh in TNBC and metastatic breast cancer was higher than that of the non-tumor and non-TNBC tissues (Fig. 1c, d). Moreover, the transcriptional activity of Gαh appeared to be enhanced in cancer tissues compared to the adjacent normal tissues derived from breast cancer patients (Fig. 1e).
To validate the clinical relevance of Gαh expression in the metastatic evolution of TNBCs, we evaluated the prognostic significance of Gαh-OE in the lung metastasis of an estrogen receptor (ER)-negative breast cancer cohort (GSE5327) from the SurvExpress database. The data showed that Gαh-OE indicated a poor probability of lung metastasis-free survival (Fig. 1f) and was notable in breast cancer tissues derived from patients with a higher risk of lung metastasis (Fig. 1g).
Gαh overexpression promotes the metastatic potential of TNBC cells
Conversely, we performed Gαh knockdown (KD) in MDA-MB-231 cells (Fig. 2i) to see if the transcriptional inhibition of Gαh could suppress the metastatic potential of TNBCs. Our data showed that silencing Gαh expression by two independent shRNA clones dramatically suppresses the invasive ability of MDA-MB-231 cells (Fig. 2j). Similarly, Gαh-KD appeared to effectively mitigate the metastatic colonization of MDA-MB-231 cells in the lungs of tumor-bearing mice (Fig. 2k–m). These findings demonstrate that Gαh-OE is likely correlated with metastatic progression of TNBCs.
Intracellular GTP-binding activity, but not extracellular transglutaminase activity, of Gαh dictates the metastatic evolution of TNBCs
To validate that the GTP-binding activity of Gαh is critical for mediating invasive activity of TNBCs, we next reconstituted the expression of wild-type and W241A/R580A mutant Gαh in Gαh-silenced MDA-MB-231 cells (Fig. 3e). Our results revealed that restoration of the expression of the wild-type and W241A mutant, but not the R580A mutant, Gαh rescues the GTP-binding activity and invasive ability of the Gαh-silenced MDA-MB-231 cells (Fig. 3e–h). Since restoring the expression of the transamidating activity-deficient W241A mutant was still incapable of recovering the invasive ability of Gαh-KD MDA-MB-231 cells, we concluded that the G protein function of Gαh dictates the metastatic evolution of Gαh-overexpressing TNBC cells.
The upregulation of cytosolic, not extracellular, Gαh predicts poor prognosis and increased metastatic risk in breast cancer
The protein-protein interaction between Gαh and PLCδ1 determines cellular invasion in TNBC cells and serves as a therapeutic target for combating TNBC metastasis
Moreover, IHC results from serial sections demonstrated that the levels of cytosolic Gαh and PLCδ1 are positively correlated (Fig. 5g). Similar to the high level of expression of cytosolic Gαh, increased protein levels of PLCδ1 was identified as poor prognostic factor in patients with breast cancer (Additional file 1: Figure S1A and Table S2) [22]. Moreover, the signature of the combination of high cytosolic Gαh and high PLCδ1 appeared to significantly predict a worse outcome in breast cancer patients (Fig. 5h). In addition, the Cox multivariate analysis of disease-free survival indicated that increased PLCδ1 level and the signature of high cytosolic Gαh/PLCδ1 were independent predictive factors of poor outcomes in breast cancer patients (Additional file 1: Figure S1B and Table S4). Notably, the high level of PLCδ1 and the high cytosolic Gαh/PLCδ1 signature correlated with higher pathologic and N stage in ER-negative breast cancer patients (Fig. 5i). These results highlight the therapeutic value of targeting the PPI of the Gαh/PLCδ1 complex in combating metastasis of TNBC.
The oncogenic role of extracellular and cytosolic Gαh in promoting cancer progression is still controversial. The increased levels of extracellular Gαh have been shown to promote breast cancer metastasis by recruiting integrin-related signaling cascades [10, 23]. Similar findings were found in ovarian cancer where extracellular Gαh promotes metastasis via activating the NF-κB signaling axis [24]. Conversely, here, we show that the increased levels of extracellular Gαh refers to a favorable prognosis in breast cancer patients. Similarly, an increased level of extracellular Gαh was previously shown to inhibit tumor invasion in TNBCs [25]. Moreover, our data showed that the loss of the transamidating, but not G protein, function of Gαh is capable of rescuing the reduced invasive ability in Gαh-silenced MDA-MB-231 cells. Similarly, it has been demonstrated that the catalytically inactive (C277S) mutant of Gαh activates NF-κB and induces HIF-1α expression as effectively as wild-type Gαh [26]. Since secreted Gαh has been thought to be responsible for catalyzing the crosslinking of ECM proteins via its transamidating activity [10], our findings suggest that extracellular Gαh likely functions as a tumor suppressor in TNBCs.
Here, we show that the pronounced accumulation of Gαh in the cytoplasm predicts worse prognosis and causally associates with the expression of PLC-δ1 in breast cancer. The loss of both the G protein function of Gαh and its PLC-δ1 interacting domain reduces the Gαh-enhanced metastatic potential of TNBC cells. Previous studies have shown that activation of the Gαh/PLC-δ1 signaling axis by several G protein-coupled receptors (e.g., α1B-adrenergic [5], oxytocin [27], and FSH [6] receptors) induces fluctuations in the levels of intracellular Ca2+ and promotes the metastatic evolution of TNBC cells by modulating their epithelial-to-mesenchymal transition (EMT), which is known as the initial step of cancer metastasis [28, 29]. Notably, the activation of FSH receptor has been shown to induce EMT by triggering the PI3K/Akt-Snail signaling pathway in ovarian cancer cells [30] and to enhance cellular migration and invasion by modulating actin cytoskeleton activity in breast cancer cells [31]. Accordingly, the activation of oxytocin receptor has been found to elevate the invasive properties of endometrial cancer cells via the PI3K/Akt axis [32] and to promote the migratory ability of prostate cancer cells by coupling to the Gi-dependent pathway [33]. These findings implicate the possible role of the Gαh/PLC-δ1 signaling axis in these events since the activation of the Gαh/PLC-δ1 pathway has been identified to elicit intracellular Ca2+ elevation, promote IP3 turnover, and modulate myosin and actin dynamics [34].
Several studies have demonstrated that Gαh-OE correlates with metastatic progression [35–39] as well as drug resistance [40, 41] in different types of cancer, suggesting that the discovery of inhibitors of PPI in the Gαh/PLC-δ1 complex could be a novel and useful strategy to combat malignant tumors. Although therapeutic targeting of the PPI of Gαh/PLC-δ1 is likely to be effective in treating metastatic TNBCs, further experiments are still needed to identify the Gαh-coupled receptor(s) and related signaling cascades in TNBC cells.
Our data delineated the inverse prognostic values of extracellular and cytoplasmic Gαh and validated that the G-protein activity of Gαh is crucial for promoting TNBC metastasis by recruiting the PLC-δ1-associated signaling axis. Blocking the PPI between Gαh and PLC-δ1 displayed an inhibitory effect on the metastatic capacity of TNBC cells in vitro and in vivo, providing a new avenue for drug discovery in cancer.
We like to thank Tzu-Ling Ting for the technical assistance.
This study was supported by Ministry of Science and Technology (MOST 104-2320-B-038-061-MY3) and by Taipei Medical University (TMU103-AE1-B13) to Dr. Yuan-Feng Lin.
Availability of data and materials
Not applicable.
Authors' contributions
SPH, CLC, and YFL contributed to conceptualization and design. SPH, PYL, CJK, CLC, WJL, and YFL helped in the development of methodology. SPH, PYL, CJK, CLC, WJL, YHT, and YFL helped in the acquisition of data (provided animals, acquired and managed patients, provided facilities, etc.). SPH, PYL, CLC, and YFL analyzed and interpretated the data (e.g., statistical analysis, biostatistics, and computational analysis). SPH, CJK, CLC, and YFL wrote, reviewed, and/or revised the manuscript. CJK and CLC performed the administrative, technical, or material support (i.e., reporting or organizing data and constructing databases). CLC and YFL supervised the study. All authors read and approved the final manuscript.
Competing interests
The authors declare that they have no competing interests.
Consent for publication
Not applicable.
Ethics approval
The experiments were approved by Institutional Review Board (IRB) (TMU-IRB 99049) of Taipei Medical University.
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Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Authors' Affiliations
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The Cadillac Break Property Group stretches for 35 km along the Cadillac Break, a regional fault zone that has produced some 100,000,000 ounces of gold since the early 1900's. Alexandria's initial 14 properties in this group came from two property agreements in late 2006 from Aur Resources Corp. (Teck Cominco) and Cambior Inc. (IAMGOLD Corp.). The Company staked or acquired the remaining properties subsequent to the initial purchase. Alexandria holds 100% interest in all 20 properties, subject to a 2% NSR, which lie in Bourlamaque, Louvicourt and Vauquelin Townships.
Geologically, the dominant feature underlying the properties is the Cadillac Break, a regional fault zone that separates metasedimentary rocks of the Pontiac Group on the south side from predominantly volcanic rocks and associated intrusive rocks on the north. However, this large property package hosts a variety of geological environments favourable to gold mineralization, including those which are known elsewhere to host large gold deposits, including: 1) shear-zone, 2) gold-rich massive sulphide, 3) porhyry gold-copper and skarn, and 3) disseminated gold.
The property group hosts four historic gold deposits: Orenada, Mid-Canada, Akasaba, and Sleepy. Two of them, Mid-Canada and Akasaba, are former producing mines, the latter having produced 40,000 ounces of gold at a grade of 5.14 g/t of gold in the early 1960's. Orenada has had underground development in the form of a 300 m shaft and two horizontal exploration drifts.
The properties have been explored by various operators since the early 1930s, with over 2,400 drill holes completed since that time. Other work includes multi-property geophysical and geochemical programs, and trenching and mapping, all of which Alexandria has been compiling into a single database. It is worth noting that recent blind discoveries in the region (as at Lapa and Goldex), at depths between 500 m and 1,500 m, show that the Cadillac Break Group of properties is highly under-explored, as most drilling has been in the upper 150 m over the whole package, only locally testing down to 300 m depth.
Please note that estimates of grade and tonnage from historical ore deposits discussed are not compliant with National Instrument 43-101, nor does the company intend to convey that these deposits are Current Resources under the guidelines defined in NI43-101. While the company views these as strong advanced stage exploration targets, with excellent potential for advancement; there is no evidence at this time that an economic deposit underlies these properties as defined in the national instrument. More work will be required, as will a thorough review of all data by a Qualified Person with a qualifying report, before these deposits can be classified according to NI43-101.


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Police are appealing for witnesses and information after a burglary on Gaydon Road in Bishops Itchington.
The incident happened between midnight on Saturday January 31 and 7.30am on Monday February 2.
Offenders forced entry to the property and stole car tyres worth thousands of pounds.
Police are eager to hear from anyone with information or who saw a dark coloured Landrover or a light coloured Iveco 35 van in the area between these times.
To give information call the investigating officer, Pc Stuart Mucklow, on 101 and extension 5480, quoting incident 32 of Monday February 2.
Alternatively, information can be given to Crimestoppers anonymously on 0800 555 111 or via www.crimestoppers-uk.org


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Mutualism of the Month: Nectar robbers and pollinating birds/
Every month we showcase a relationship between two or more species or groups of species that can be considered a mutualism: a relationship where both members benefit. This month's mutualism is between the nectar-robbing purple sunbird Nectarina asiatica and a small flowering tree, the desert teak Tecomella undulata, particularly how the purple sunbird impacts the relationship between desert teak and its two pollinating birds: the red-vented bulbul Pycnonotus cafer and the white-eared bulbul P. leucotis.
Nectar robbing refers to the act of an animal removing nectar from a flowering plant without pollinating the flower. Typically, this phenomenon occurs with flowers that conceal their nectar in a tubular corolla (botanical term for all the petals) [picture]. The animal pierces through the corolla tube to remove the nectar, incidentally, never coming in contact with the reproductive parts of the flower. Pollination of these flowers usually requires that animals (typically birds or insects) touch the male anthers of the flowers, picking up pollen and then depositing the pollen grains on the female stigma of other flowers.
The term nectar-robbing confers a negative connotation: the nectar-robber steals nectar at the detriment of the plant; this isn't always the case. The positive effects become obvious when fruit-set (the amount of fruit produced by a plant) increases in response to nectar-robbers: a sign that nectar-robbing increases reproductive success.
How could this be?
By damaging the floral parts of the tree and removing the nectar within the flowers, nectar robbers can indirectly alter the behavior of pollinators.
This is what happens in the three-way relationship between the purple sunbird, the desert teak, and the bulbuls. Sunbirds can pierce more than half of the flowers of a desert teak, removing the nectar from the flowers and frustrating the bulbuls. Although nectar is replenished regularly, insufficient amounts of nectar compel the bulbuls to visit additional trees in their search for nectar. This is great for the desert teak as they are self-incompatible; their pollen cannot fertilize their own flowers. Pollinators must bring the pollen to another tree.
Scientists discovered this gem of information out by mimicking the sunbird. They drained all the flowers in eight desert teaks (1296 flowers!) with a small syringe and protected another eight trees (1120 flowers!) from robbing by wrapping the flowers in cellophane tape. They recorded how many flowers were visited by each Bulbul and how many subsequent trees were visited. Bulbuls that visited nectar-drained teaks visited significantly less flowers per bout (5 vs 19), but visited significantly more trees per bout when they couldn't find nectar (7 vs 2). This should increase pollination in a self-incompatible species like teak and it did! Robbed trees had a significantly higher fruit-set. So while bulbuls have to work harder, reducing their benefit in this mutualism, the sunbirds and teaks benefit greatly from this relationship.
To read the original scientific article from the journal PLOS One click here.
Don't forget to check out more mutualisms of the month and all the other great articles on FTDM! Stay hungry!


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Skegness & District runners embarked on their now annual relay, taking in all the town's that are in the area the running club covers.
This year's relay saw the runners wearing hula skirts - pictured - as well to add to the carnival atmosphere of the event.
More than 40 club runners took part with Andy Shelton and Stuart Cragg getting the relay going at 6am.
The route travelled from Skegness through Wainfleet inland to Midville, down to the Hobhole on the quiet back roads through to Witham Country Park in Boston.
It was then up towards Woodhall Spa and Horncastle, to Spilsby then back through Burgh le Marsh and into Skegness for a welcome supper of fish and chips, just more than 12 hours after starting.
Shelton and Cragg both ran more than 30 miles on a route of about 75 miles in total.
Sarah Coupland stretched her legs to record more than 22 miles and Helena Shelton managed 25 miles.
The relay was a great way to get the club's newest runners involved too as the legs were distances from three miles upwards.
There was double club success for Skegness & District at the Newark Half Marathon.
With 12 runners representing their club, two trophies were won.
Stuart Cragg was second in the Vet40 class in a time of 1.19:28 and Colin Chambers once again collected the prize for first vet70 in a stunning time of 1.43:55 .
Skegness and District Running Club are continuing to support Ben Smith's 401 challenge, as he attempts to run 401 marathons in as many days to raise awareness and funding for antibullying charities.
Member Kim O'Connell joined runners at Clumber Park, near Worksop, for Ben's marathon number 348, and she managed her furthest distance to date, just more than 22 miles, which was a great achievement having taken up running around a year ago.
Stuart Cragg and Melanie Standbrook travelled to Ladybower Reservoir for marathon 350 with runners from around the area.
Both Stuart and Melanie ran the full marathon distance, after Ben visited Skegness last December and inspired members.
Stephen Bark had a fabulous run in the Sedbergh Hills Fell Race, a technical 14 miler with and ascent of 5100ft.
The route was a bit of a roller coaster through the hills and valleys, but the dry weather had made the trail a very runnable surface which helped.
Stephen, representing Skegness and District RC, finished in 3.14:02 which beat his previous best by 50 seconds.
Feeling strong in the second half of the race, Stephen was 37th from 96 starters.


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Subject: Expanded Examination Cycle Eligibility
Date: January 6, 2017
To: Chief Executive Officers of All National Banks, Federal Savings Associations, and Federal Branches and Agencies; Department and Division Heads; All Examining Personnel; and Other Interested Parties
Description: Final Rule
The Office of the Comptroller of the Currency (OCC), the Board of Governors of the Federal Reserve System, and the Federal Deposit Insurance Corporation have published an interagency final rule that adopts without change the February 29, 2016, interim final rule amending the regulations governing eligibility for the 18-month on-site examination cycle. Pursuant to the Fixing America's Surface Transportation Act (FAST Act), the interim final rule made qualifying 1- and 2-rated national banks, federal savings associations, and federal branches and agencies (collectively, banks) with less than $1 billion in total assets eligible for an 18-month (rather than a 12-month) examination cycle.
Note for Community Banks
Qualifying 1- and 2-rated banks with less than $1 billion in total assets are eligible for an 18-month examination cycle.
Prior to the interim final rule, 18-month examination cycles were available only to qualifying 1- and 2-rated banks with less than $500 million in total assets. Through the interim final rule, the OCC revised 12 CFR 4 to raise the threshold for qualifying 1- and 2-rated banks with less than $1 billion in total assets, consistent with the FAST Act and section 7(c)(1)(C) of the International Banking Act of 1978. Other qualification criteria remained the same.
Specifically, the interim final rule provided that to qualify for the extended 18-month examination cycle, 1- and 2-rated banks with less than $1 billion in total assets must be well capitalized, not be subject to a formal enforcement proceeding or order from a federal banking agency, and not have experienced a change of control in the preceding 12-month period. Additionally, a national bank or federal savings association must have a management rating of 1 or 2 to qualify.
Under the interim final rule, the OCC retained the authority to maintain the current 12-month on-site examination schedule for an institution, or adopt a more frequent schedule than every 18 months, if the agency determines it would be necessary or appropriate.
The final rule adopts each of these provisions without change.
Further Information
Please contact Deborah Katz, Assistant Director, or Melissa Lisenbee, Attorney, Legislative and Regulatory Activities Division, at (202) 649-5490; or Scott Schainost, Midsize and Community Bank Supervision Liaison, Midsize and Community Bank Supervision, at (202) 649-8173.
Amy S. Friend
Related Link


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The center of energy-informational medicine
"Center of energy-informational medicine" was founded in 2001 and became one of the first medical facilities in Samara, working in the field of bioresonance medicine. At the present time in different parts of the city are four branches of the Center, preparing the opening of the fifth.
The main direction of CAIM – computer electropuncture diagnostics and bioresonance therapy.
Highly qualified specialists using modern diagnostic techniques identify deviations in health status at the stage of preexisting diseases, determine the causes of certain diseases, and based on these data, the individual program of treatment.
The appointment is conducted in a comfortable environment – the test procedure is painless, does not require the introduction into the body in a short period of time the patient receives a large amount of information about your health condition and doctor's recommendations for treatment and health improvement. According to the results of diagnostics conducted a session of bioresonance therapy.
For 8 years the specialists of "Center of energy-informational medicine" has developed a number of software copyright:
program "Life without drugs" provides a comprehensive survey of the body comprehensive therapeutic programs without the use of pharmacological agents.
the program "Eat and grow thin" will help everyone who is interested in the problem of excess weight. Spetsialistidele causes of metabolic disorders, make an individual Wellness program, choose products, accelerating the metabolism for getting rid of excess weight in the comfort of the body. Continue reading
Bio-energy therapy (Energy healing)
Managing the energy of the human biofield, as discussed in the article "bio-energy impact", allows you to perform diagnostic and healing steps.
Briefly describe some moments of this practice.
Some of the techniques of diagnosis
Each cell of a living organism gives a whole range of signals: sound, color, vibration, temperature. The ability to recognize these signals in combination with in-depth knowledge of anatomy, physiology, allows a maximum precision to carry out medical diagnostics.
Touch diagnostics is an important addition to the hardware and laboratory diagnostics. Is it as auras directly on the basis of tactile sensations of the hand with a framework of indicators or pendulum, internal, or mental vision.
When conducting autodiagnostika it should be borne in mind that the form of the aura of a healthy person is round or oval, ovoid, smooth; tactile sensations – tight, elastic. If there are any abnormalities in the body shape of the aura varies, as well as the sensation. It is expressed in the emergence of the so-called energy pits, craters, humps and breaks.
Another common type of sensor diagnostics – diagnostics by the chakras. As already noted, the human biofield is maintained at a particular desired level of the energy centers – chakras.Ancient and modern researchers have found that the chakras are conventionally conical shape or the shape of a Lotus flower with a funnel located at the front just below the skin and the top few millimetres of the spine. Each chakra is correlated with a specific group of organs. Violation of energy of the particular chakra indicates a predisposition relevant group of bodies to a functional or organic disorders. Diagnosis can be performed using the framework of indicators. Continue reading
How animals heal humans
Animals are not only loyal people, they know how to treat. It has long been known that cats and dogs are able to relieve the stress of the owner, to normalize the pressure, and with horses and dolphins can cure even mental diseases. But the "animal" therapy choice deliberately, to attract professionals or closely monitor your pet. After all, the same cat, for example, may not be in the mood to treat you…
Animal-assisted therapy – the treatment of animals is shrouded in myths. Medical literature likes to describe this process literally as a panacea, almost comparable with the effects of traditional medicine. Unfortunately, failure to provide medical assistance, hope for a positive outcome in the application of alternative methods may lead to irreparable consequences. And such cases already described quite a lot. Therefore, it is better to understand the purpose of animal therapy as a psychological "crutch". support the person in a difficult situation for him.
Thus, dogs often "work" with children with autism and those with down syndrome, patients with cerebral palsy. No matter whether it's a cute little dog Maltese or reliable, faithful friend Labrador, in any case, socializing with friends person particularly well in locatedhere psychophysiological development. problems of communication when the child is aggressive or, on the contrary, reserved and shy. And this is a great work for the dog. Because sometimes she just needs to lie still with your sick hours so that he relaxed and began to trust her, sometimes crawling with the child or to overcome obstacles. And sometimes she even obliged to endure shoving, punching, loud cry of a baby – in General, any behavior that goes beyond. However, she has no right to growl, to bark, growl or otherwise show their displeasure. Continue reading


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My current series is called "becoming" and depicts girls between 12 and 13 years old.
I find this to be a fascinating time as it marks that important transitional period from childhood to adolescence. I am attempting to catch those last moments of innocence before that sense of "knowing" arrives. While my charming models have invariably shared a self-confident persona with me, I recognize that they are also volatile and unpredictable. They can be mature, affectionate and delightful, but also self-centred, moody, surly and backtalking. As this is a liminal stage when the individual is "neither here nor there; they are betwixt and between" (Victor Turner) I will be including portraits where the model shows a pensive, thoughtful moment, or is being goofy and silly as well as images of the girls being competent and powerful. Being a teenager has never been easy and I wish to portray some of the inconsistencies I have noted as these young women deal with the challenge of feeling invincible and invisible at the same time.
Project Date
June 2016 to the present


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"Knowing is not enough; we must apply. Willing is not enough; we must do. I have come to the frightening conclusion that I am the decisive element. It is my personal approach that creates the climate. It is my daily mood that makes the weather. I possess tremendous power to make life miserable or joyous."
Johann Wolfgang von Goethe (1749 - 1832)
What free enterprise words are these: "I am the decisive element."
That says it all!
That defines what freedom, and individual liberty, and endowed rights, and economic self interest, and personal leadership, and individual development, and free-market capitalism, and private property are all about!
Private property? Yes, we are our own private property. We improve ourselves, maintain ourselves, we add and subtract value, and grow or diminish ourselves!
And Goethe arrived at that point and found that "frightening conclusion" to play a huge part in his personal success or failure.
Thus it is with free enterprise. We can be our own best asset or our own worst enemy. To the extent that we improve (by ourselves and with the help of others) our domain, or our product, or our service, or our abilities, we add value and assets.
When humans are interested in something they tweak it! I tweak my blogs from time to time. I tweak my website often, almost daily! The people who create and manage the apps on our phones are continually offering updates!
That is what free markets do. And they do it automatically. There is no overlord directing the ebbs and flows of businesses large and small. The "Invisible Hand" of the free marketplace is on autopilot, and each of its biggest and smallest parts are doing their own things in response to its direction.
So, as Goethe suggests, knowing is important, but different than applying. Both must be done.
Wanting and willing to do is not enough; we must be willing to do.
And the power of free enterprise rests within us.
So, let us go! Leave us to do!
Let those who are the most knowing get to the successful applying!
Let those who are the most willing and get to the successful doing!
Let those who are the most desirous to experience joy be successful in creating it!
Goethe's conclusion is that we need to realize who we are and then apply ourselves.