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What is the Thunderbolts Adventure and when will it be held?
The 2019 edition of the Thunderbolts Adventure will be run on 13 – 14 April 2019. It will be an overnight gravel adventure in the Barrington Tops in New South Wales designed for competent cyclists who are experienced in long distance riding.
You don't have be be a superfit racing machine to enter this race, but you do need to:
- Understand how to fuel your body for an extended ride covering over 100 km on remote gravel roads
- Be experienced at navigating and following a GPX route
- Have a reasonable level of mechanical know how would you need to perform running repairs on your bicycle eg, punctures, ripped tyres, broken chains, non-functioning gears.
Is it a race?
No. But kind of. A few will race for line honours and Strava bragging rights, but most will race themselves and look to team up with a small bunch of riders of similar ability.
Whether you are racing yourself or others there is a chance you could be riding by yourself at times, so it is most important that you come fully prepared and self-sufficient. Better still bring your mates who you normally ride with and you are sure to have a great ride in spectacular country with friends.
Is there support?
Plan to be self-sufficient. You need to navigate your way along the course as there will be no event markings. You will need to carry your own nutrition and hydration. You need to carry sensible protective clothing keeping in mind that weather conditions can be quite changeable. You need to carry any spares that you feel you might conceivably need on the day. There will be 1 opportunity to top up bottles on each day.
- There is a water top up point at around the 60 km mark on Day One.
- You will be able to leave some water bottles with a vehicle which will deliver them to a designated location around half way through the ride on Day Two.
We will have a sweep car drive through after the event on both days, to make sure everyone is back safely, but you could be waiting a long time for this car to come through and it may not have space for you so don't rely on this as a sag wagon. It is primarily to ensure the safe passage of the riders.
Who will be doing support?
Great question. We are looking for people to help out. Give us a shout if you know someone or would like to volunteer yourself. We will cover your expenses and we reckon there is a good chance you wont have to buy a single drink on Saturday night.
Are there checkpoints?
You will need to sign on at the start of the event. We will have a sign in "Check Point" at Victoria Hotel Moonan Flat Pub at the end of Day One – the second "Check Point" will be at The Roundabout Inn in Gloucester on Day Two.
You will need to sign into both locations as you pass through so we know what section you are on should something go wrong.
What are the route options?
The event is going to be a total of up to 250 kms with approximately 5000 vm of elevation over two days. You'll traverse the entire Barrington Tops and return to the start point in Gloucestor via a mostly remote gravel roads.
This is an Overnighter that will be a tough but enjoyable two days – the event in previous years was completed in one day.
When will I get the route?
We will send you the route a couple of weeks before the event. We will send you a Strava route and GPX file. How you navigate is up to you. If you want cue cards, paper maps, topos etc that will be up to you to get.
We expect every individual rider to be able to navigate the route. You may not be riding in a bunch and this is a bloody big remote area.
What do you mean by remote?
The majority of this event takes place a long way from civilisation. There is little to no traffic on these roads and no facilities. Except for Victoria Hotel Moonan Flat Pub there are no shops along the route. There are very few residents to be able to assist you. Of course, we, the organisers will do what we can to help should you be unable to continue and still a long way from the finish but having a PLB or SPOT Tracker is highly recommended.
Is there phone coverage?
Not really, apart from in Gloucestor. We recommend you do not rely on your mobile phone for navigation or communication.
Do I need a SPOT or PLB?
You are riding at your own risk. If you have a SPOT Tracker that would also be adequate (but note that SPOTs are not as useful in an emergency as a PLB which will directly contact AUSAR). If you don't have one, hire, borrow or buy one. Eg. http://www.epirbhire.com.au/ or Blue Mountains Police Rescue and some NPWS.
What bike do I need to take?
The route is perfect for a Gravel Bike or CX – consider running the biggest rubber that will fit your stead will fit – run at least 38-40c tyres with at least some tread (ie. not slicks).
Please don't bring your roadie, even if you have 28s on it. The Barrington Tops road is hard with large chunks of embedded rocks. A mountain bike would be fine, but it's a long way to ride on a MTB.
There are two big climbs. This first climb is around 1000 vm and the second is about 800 vm which gets over 15%. Low gears would be nice. Some highly credentialed riders ran 42 F x 32 R (ratio 1.31 with 29er Wheel) as their lowest gear last year and found that quite challenging.
What is the quality of the roads?
The roads are all good quality gravel roads with very limited bitumen. Barrington tops rd is very compacted gravel with large embedded gravel which can be tiring to ride. There is hero dirt and lots of smooth gravel as well as some more washed out gravel roads.
There are two sections where extra care should be taken. The descent on the western side of Barrington tops is smooth and fast. It also lack guard rails at many points. In addition these is a very steep and loose descent on the northern return section.
What spares do I need?
Everything to get you through the ride, but if your bike is well maintained then punctures are the main concern. Keeping in mind that there are no bike shops along the way, consider your worse case scenario and plan for that. For example, let's say you have been unlucky and had a few punctures. You are on your own, you are out of tubes and cannisters. If you have a hand pump and a some patches you might be able to repair a tube to get you out of trouble. A $5.00 note is always handy for blocking a torn sidewall.
Other spares you might consider would be a chain breaker, a spare quick link and a spare Derailleur hanger.
In addition to this consider bringing a basic first aid kit and space blanket.
What if I want to pull out of the race?
Don't count on anyone picking you up. We will have a sweep vehicle, but there may not be space on it. The locals are friendly – we had a mate who was lucky to get a lift back to Gloucester last year!
Do I need lights?
Yes. While you'll likely be back before it gets dark – these are a safety feature so please bring at least one rear flashing red light and a solid white front light with 4 hours capacity.
Is this a sanctioned race?
No. You ride at your own risk. We highly suggest you have ambulance cover. The roads will not be closed. The route will not be marked. We are sure other riders will do what they can to help you, but in the end you're on your own, and responsible for yourself.
Is there a KOM?
There maybe a few sneaky segments out there with some prizes at the finish for various achievements. To be eligible you must log your ride on Strava (and have it uploaded by the time we have a beer in the pub at the end of Day Two).
What is cost of the event?
Dinner on Saturday night and breakfast Sunday morning will be catered for and included as part of the event entry fee. We will also be providing a little bit of event schwag, reimbursing expenses for the support vehicles as well a funding the ongoing development of the Graveleur Community.
The total cost per rider will be approx $ 70.00 and entries will be limited. More information about how to secure your place will be available soon.
Camping will be no charge Victoria Hotel Moonan Flat Pub grounds, however if you want a warm bed at the Pub book now. We have arranged a special rate, just mention you are part of the Thunderbolts Cycling event
While the beer is cold (as well as other beverages) – all drinks will be at your own cost.
Will I need to carry my tent?
If you are planning to camp, we can get your tent/swag/sleeping bag dropped at Victoria Hotel Moonan Flat Pub – one less thing to carry on your bike.
This will be returned to Gloucester on Day Two.
If you have any other queries, please email us at email@example.com


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Easy Brackets or Cs4 quick release mounting brackets make saddlebags detachable.
Easy to install ! Lockable !
Easy Bracket docking posts are model specific
Cs4 docking posts are more universal
Prices include 21% VAT.
Nationals of countries outside the EC (European Community) do not pay this VAT.
The VAT will be deducted automatically when you go to "checkout" and you fill in the shipping location.

subset: commoncrawl


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There's only one way to spell NO.
Posted on 16 January 2019
Sometimes when I'm writing and I'm really passionate / fired up about something, my brain moves too fast for my fingers. I lose track of my thoughts easily and I can't keep up with myself. Runaway train, literally.
This subject is getting to be more and more of an issue for me. I think it's because I am fully aware of it now. VS. having fell a victim to it or being trapped by it in the past-- but regardless, I'm going to do my best to use my words. Because it's not okay and I want to talk about it.
I think most of us can agree that we're all, in some way, by any and many definitions of the word, seeking love.
SIDEBAR: If you don't agree with that statement keep reading, it's not my point.
I, along with people who have a beating functioning heart (my last 2 exes excluded) look for an emotional connection. Something deeper, something that feels real. Romantic, platonic, a bird you talk to outside your window (I do that), something or someone that makes you feel good. Can we agree that love or a connection, in it's most general definition, is something we all crave?
Okay. So with that said, in the current state of affairs aka the "hook up culture"
SIDEBAR: I hate that term. If I can think of a better one while I'm writing this I'm going sidebar it in whenever I do.
or whatever the F you want to call this broken down, 2 wheeled short bus that I see dating as right now-- it could be difficult to find someone with the same values when it comes to relationships. Not that I'm actively looking or currently interested in even holding more than a few texts with someone here and there, it ain't easy. To simplify it even further, people just don't know how to act. They rarely did, but it's truly fading faster and faster.
To me, this "hook up culture" is what has directly effected people's approach to dating. Not only their warped definition of what dating is, I mean their literal approach to another human being. Call me old school, but half assed attempts, hit-em and quit-em dating apps and DMs ain't where it's at.
SIDEBAR: Listen, I'm not saying that you can't just sleep with someone and be done with it, to each his own. I'm also not saying that everything needs to be headed to the alter or nothing at all-- but this casual shit is not for me. At this stage in my life, I'm not sleeping with you unless I love you. Period. Do I sound like a Puritan? GOOD. It's been over a decade since "that wild summer in my 20s where I had a lot of fun at D'jais" so I'm obviously good. Besides, you should be treating your body like a temple, not a fucking trash can but that's a WHOLE 'nother story.
SIDEBAR PT2: Second disclaimer, because; no trolls. Let me just say right now that I do not hate men. I don't hate anyone for that matter. And I am not talking about all men right now. I'm solely speaking from my own recent experiences in hopes to connect to others experiencing the same thing. If this offends you, you're the problem.
Lately, I've find myself in conversations with guys who:
1. Are unable to correctly identify emotions and take responsibility for them.
2. Instantly throw a bitch fest and a hissy fit when their mediocre attempts are not met with the response they're expecting or think they deserve.
SIDEBAR: Rule #1-- Don't expect shit from anyone. No one owes you shit.
3. Have shitty intentions and try to manipulate people to get what they want.
Yes, while points 1-3 are all irritating and extremely aggravating, for me, they do not slightly compare to the rage I feel when #4 is not respected.
And this isn't my first time touching on the situation.On August 21st, I posted this:
And this was was my caption:
Disclaimer: I usually do not engage with blatant idiots but somebody had to be the example. THE STORY: Today a complete and total stranger (on my private acct) told me, amongst other rude comments out of nowhere, to FUCK OFF bc I wouldn't give him my number and or go out with him. After me politely telling him I wasn't interested WEEKS AGO, he felt the need to keep asking, badgering and throwing himself a pathetic little pity party all alone in my DMs as if I would feel different and or sympathetic? NOPE. You're wrong. Wrong girl. Not only did I put your chat on silent WEEKS AGO, you turned me off further when I wasn't even interested to begin with. Lines and lines of unreplied messages is a sign you're in this alone, bro. I can tell you aren't that bright but do you realize that? Which brings me to my real point... What about the word NO is so hard for people to understand? And why am I the bad guy for simply not being interested? Furthermore, who the fuck do you think you are taking out your obviously sick personal baggage on a complete stranger? I chose to not tag this person bc I feel as though their life is most likely embarrassing enough but let me make something very clear... I don't care if it's a text or God forbid something physical, NO MEANS FUCKING NO. One more time for the cheap seats in the back, NO MEANS NO! So homeboy if you're reading this, YOU'RE AN ASSHOLE. How's that for me being "the worst"?! Oh yea, and I'm 32. Get out of grown woman's DMs and go out and meet human beings like a normal person. | #nope #boybye #blocked #rant #greevingcards
I am sick to death of the guilt trip laid on a female that comes after denying a guy. It is NOT OKAY for you to put me down after you fail to understand the simple truth. If I turn you down-- be it repeatedly, after realizing I'm not feeling it or from the jump-- I do it kindly, honestly and with respect for other peoples feelings. I get it, no one likes to be rejected... but be the fucking man you claim you are, take the L and keep it moving. More importantly, how you take it is your problem, not mine.
Nowhere in the Handbook of Life does it claim that you should take a shot at someone just because your ego got burnt. Furthermore, if you had listened to or respected my feelings from the first time you asked me out, we wouldn't even be here. Especially considering I have never, EVER given you any reason to think otherwise.
There has been someone in my life who has been trying to make it happen with me for years. An acquaintance of mine that wasn't always on his bullshit but I mean, trying for YEARS. So long that it's straight astonishing he is even still trying. Not only have I been in relationships, which he has known about and respected at the time, I generally am just not interested in the fucking guy- nor will I ever be. And he has heard that answer plenty of times before.
The fact that I'm not interested? THAT IS OKAY. THAT IS MY RIGHT AND THOSE ARE MY FEELINGS. Nowhere in the Handbook of Life does it also say that you have to give a chance to everyone that likes you and thinks they are worthy of the job. That'd just be bizarre. You like what you like and you want what you want. Plain and simple. You shouldn't have to explain your reasoning to anyone.
Why does everything have to be defined anyway? I said no. NO IS THE SHORTEST WORD IN THE DICTIONARY THAT CAN BE USED AS A COMPLETE SENTENCE. There is nothing else to say. The conversation begins and ends with that word. This isn't "let's make a deal". Or better yet, that other show-- "let's beat her down till she caves".
SIDEBAR: Idk if NO is really the shortest word in the dictionary that can be used as a complete sentence but for the purpose of this discussion, it is. Sounds like it could be true? I will get back to you on that.
SIDEBAR PT2: I'm also fully aware that everyone has a different copy of said Handbook of Life. And that some people have no Handbook, just a half a brain and some bad ideas. Also that this Handbook I keep referencing doesn't exist.
SIDEBAR PT3: Wouldn't it be a lot easier if there was a basic Handbook everyone followed? Written by me? Hmm...
Back to the story-- For the 5,923,562,087th time in my life, last weekend he approached the subject. Via DM (insert eye roll and teeth suck) and he asked me when I'm going to "come to my fucking senses" and that he "doesn't understand what's wrong with me".
3. WHAT'S WRONG WITH ME? What makes you think there is something wrong with me? Because I'm not interested in you? OH RIGHT, 4 YEARS LATER, THAT MUST MEAN I'M THE ODDBALL WITH A PROBLEM.
That is literally not the way this shit works.
I then told him in English, word for word as follows; "I DO NOT WANT TO DATE YOU. NOT NOW, NOT EVER. HOW ELSE CAN I SAY THAT? SO STOP IT." He proceeded to say a few unfavorable remarks and ended with telling me to "GET OFF MY HIGH HORSE."
So let's all get this straight. It's okay to now knock me down because I said no? All of a sudden I ain't shit and am now riding a high horse? But 4 years and 2 minutes ago you wanted me? Right? I was just checking.
And that was my exact response.
He had nothing to say after that. A while later I laughed and blocked him. Because if I didn't laugh at this shit in my life, I'd murder everyone.
Now had this happened to a past version of me? It would have effected her way differently.
For starters, she didn't have a platform to talk about it.
All GaGa's aside, past versions of me had way less confidence. She was a little clueless to the manipulations of life and blindly trusted everyone equally. She sometimes ignored bad vibes for mediocre attention even if it was a part of a situation that didn't flow naturally. She was honestly a little desperate and pretty naive. She would have totally second guessed herself down into a negative thought spiral.
"Maybe there is something wrong with me?" "Maybe I should just go, he must really like me if he keeps trying?" "Maybe he is a nice guy" "Maybe I need to apologize?" "Damn, now I feel like a piece of shit."... and so on and so fourth.
I would have made plenty of excuses for his insecure and generally ridiculous behavior all while taking responsibility for someone else's ludicrous feelings and unjustified response just because I spoke my truth. Then I would have felt bad about "being the bad guy" even though I had already said no years ago.
But, when it comes down to it-- he's not a nice guy. He's disrespectful and selfish with a disturbingly large sized ego. Three qualities in capital letters on my GTFOH List. He insulted me and my integrity because he couldn't handle the truth.
I guess what I'm getting at is this...
I not that girl.
I am not the girl you think you're gonna break down.
I am not the girl who will eat your disrespect and let it make me second guess myself.
I am not the girl who is going to sit back and not call you on your shit.
I am not the girl who is answering your DMs or who you'll find on a swipe for a quickie.
I am NOT THAT GIRL.
If you're looking for someone like that, keep walking.
SIDEBAR: After an extremely brief Google search, it doesn't say anywhere that NO isn't the shortest sentence so I'm sticking with my theory. Smart AND articulate. Learn something new every day. #GOALS.
1 comment
Daniela Amini: September 17, 2019
I know this is a post from January but I happened to come across it just now and can I just say how much you just totally RULE?!?!?!?!? I'm not surprised since I routinely find myself cracking up at the wit and out-of-the-box wisdoms of your cards and other products but this…THIS…is just so well said, all of it. In between snot-running-down-my-face-laughing I would pause and manage to spasmodically eek out the words "TRUTH!" before falling backwards and clutching at my sides in hysterics again. Just everything you wrote, from the overall dismal effects of the "hook up culture" to the brevity of how you dealt with potential haters to the survey of typical behaviors and the way you express yourself overall…soo, so good. xoxoxoxoxoxoxoxoxoxoxo


text:
Search for positions by entering key words in search box below. Use left side navigation to filter by Region, Job Category, Job Type or Salary.
The actual salary shall be determined by the final candidate's qualifications and experience. In addition to the starting salary, employees in the Office Specialist II classification shall also receive an approximate five percent (5%) ongoing non-pensionable compensation pay.
Office Assistant I/II (19952530)
Applications accepted until Wednesday, January 8, 2020 OR upon receipt of the first 150 applications- whichever occurs first
Written Exam: Thursday, January 16, 2020
Oral Exams: TBA
Office Assistant I: Under direct supervision, the Office Assistant I performs a variety of technical office work and other related work as required.
This exciting position supports the mission readiness of Menlo Park Fire's Urban Search and Rescue Team (US&R), Task Force 3 (CA-TF3). The successful candidate will have a keen attention to detail, be solutions oriented, and have an exceptional work ethic. This position is critical to ensuring the team is deployable to respond to disasters when called upon. As such, this person may be called upon in the evenings or on weekends in cases of emergency or national disasters.
Human Resources Specialist (19952146)
Applications accepted until minimum number meeting our qualifications have been received.
Interviews: TBA
Under general direction, performs complex and responsible technical and clerical work for the Director of Human Resources in the administration of the City's Human Resources program. Completes related work as required.
CLOSE DATE: Friday, January 3rd, 2020
II – $29.50-$35.85 Per Hour
III – $29.95-$36.40 Per Hour
(New employees should anticipate being hired at the beginning of the salary range.)
Degree: 15% - LCSW or MFT License.
A complete application packet must be received in the Glenn County Personnel Department by
CLOSE DATE: Friday, January 3rd, 2020
I – $16.31-$19.83 Per Hour
II – $18.47-$22.46 Per Hour
(New employees should anticipate being hired at the beginning of the salary range.)
A complete application packet must be received in the Glenn County Personnel Department by 5:00 P.M., Friday, January 3rd, 2020
Human Resources Technician (19949469)
The City of St. Helena has a challenging and exciting career opportunity to fill a Full-time position in the Human Resources Department. This is a great opportunity to become part of a team of dedicated individuals that take pride in serving the local community.
Human Resources Technician*
Salary Range: $59,343—$72,132/year
($28.53 - $34.68 hour)†
The Position:
This is a full journey level position. Perform a wide variety of responsible bookkeeping and clerical duties under general supervision and within a framework of established procedures with only occasional assistance or instruction. Perform a large amount of the daily bookkeeping and administrative work associated with cash & investments, accounts payable, accounts receivable, payroll, and the general ledger.
Examples of Job Duties:
Green Facilities and Operations Analyst (19949102)
Date Posted: December 5, 2019
Classification: GREEN FACILITIES AND OPERATIONS ANALYST
(This position is grant funded through December 31, 2020.
Continuance in this position is contingent upon additional funding beyond
December 31, 2020)
Open recruitment
The City of San Mateo Community Development Department is looking for an
Executive Secretary to the City Attorney


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IPL 2018 (Match 23) Preview: Mumbai Indians vs Sunrisers Hyderabad
Mumbai, April 23 (CRICKETNMORE) Sunrisers Hyderabad will aim to recover from the recent slump and get back to winning ways when they face an under-pressure Mumbai Indians in a league encounter of the Indian Premier League (IPL) at the Wankhede Stadium here on Tuesday.
Sunrisers have slipped from a comfortable three-match winning start, suffering back-to-back defeats to Kings XI Punjab and Chennai Super Kings (CSK) respectively, and would be keen to bounce back.
Also Read: We backed Nitish Rana from the beginning and assured him support, says KKR coach Jacques Kallis
Defending champions Mumbai, on the other hand have a tough task to rediscover themselves after slumping to their fourth loss in five games.
The two team will face each other in the twenty-third game of IPL 2018 tomorrow i:e Tuesday at 8 PM IST.
Key Players
Sunrisers Hyderabad - Kane Williamson, Yusuf Pathan, Rashid Khan
Mumbai Indians - Rohit Sharma, Ishan Kishan, Krunal Pandya
IPL: Head to Head
Matches - 11, SRH Won - 6, MI Won - 5
Match Prediction (POLL)


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Angiotensin II receptor type 1 blockers (ARBs), widely used antihypertensive drugs, have also been investigated for their anticancer effects. The effect of ARBs on prostate cancer in experimental models compared with meta-analysis data from clinical trials is conflicting. Whereas this discrepancy might be due to the use of supratherapeutic doses of ARBs in cellular and animal models as compared with the clinical doses used in human trials, further investigation of the effects of clinical doses of ARBs on prostate cancer in experimental models is warranted. In the current study, we sought to determine the effects of candesartan on prostate cancer cellular function in vitro and tumor growth in vivo, and characterize the underlying mechanisms. Our analysis indicated that clinically relevant doses of candesartan significantly inhibited growth of PC3 cell tumor xenografts in mice. Interestingly, the same concentrations of candesartan actually promoted prostate cancer cellular function in vitro, through a modest but significant inhibition in apoptosis. Inhibition of tumor growth by candesartan was associated with a decrease in vascular endothelial growth factor (VEGF) expression in tumors and inhibition of tumor angiogenesis, but normalization of tumor vasculature. Although candesartan did not impair PC3 cell viability, it inhibited endothelial-barrier disruption by tumor-derived factors. Furthermore, candesartan significantly inhibited expression of VEGF in PC3 and DU145 cell lines independent of angiotensin II type 2 receptor, but potentially via angiotensin II type 1 receptor inhibition. Our findings clearly demonstrate the therapeutic potential of candesartan for prostate cancer and establish a link between ARBs, VEGF expression, and prostate tumor angiogenesis.
Prostate cancer is the most commonly diagnosed cancer among males according to the American Cancer Society (Siegel et al., 2012). About 68% of prostate cancer cases are diagnosed in the 55–74 year age group (Siegel et al., 2012), and this age group is also characterized by the high prevalence of comorbid conditions, most notably cardiovascular diseases (Roger et al., 2012).
Recently, a number of meta-analyses investigating a possible link between cancer incidence and cardiovascular disease drugs have been published (Sipahi et al., 2010; Mearns, 2011). One of the major targets of these analyses was the angiotensin II receptor type 1 blockers (ARBs), which are commonly prescribed for the management of cardiovascular diseases. The results of these analyses were controversial, with some suggesting a causal link between cancer (Sipahi et al., 2010) and ARBs, whereas others dispute such a link (Mearns, 2011). To further complicate the matter, there is a plethora of experimental evidence that suggests a possible beneficial role of ARBs in the management of multiple types of cancer, especially urogenital cancers (Miyajima et al., 2002; Kosaka et al., 2007; Takahashi et al., 2012). Experimental data demonstrated the ability of ARBs to inhibit progression and metastases in bladder, renal (Miyajima et al., 2002), and prostate cancer (Kosugi et al., 2006; Kosaka et al., 2007; Takahashi et al., 2012). This beneficial effect has been consistently reported both in monotherapy settings (Kosaka et al., 2007) and in combination with other antineoplastic agents. The antineoplastic effects of ARBs are believed to be due to their ability to inhibit cancer angiogenesis (Kosaka et al., 2007), which has been shown to be associated with severity and metastatic potential of prostate cancer (Kosaka et al., 2007).
Despite the solid experimental evidence supporting an antineoplastic effect of ARBs, the controversy between clinical and experimental data must be resolved. In the majority of experimental studies, the dose of ARBs used is supratherapeutic and always in combination with angiotensin II (AngII), which cannot be extrapolated to clinical practice (Uemura et al., 2003, 2005a; Takahashi et al., 2012). This point has been critically reviewed, and the importance of using clinically relevant doses of pharmacologic agents in preclinical studies has been noted (Reagan-Shaw et al., 2008). Another important consideration in investigating the effects of ARBs is the concomitant treatment with exogenous AngII (Uemura et al., 2003; Kosaka et al., 2007; Chen et al., 2013), which only blunted AngII-mediated effects. This paradigm ignores AngII-independent effects of candesartan as well as the role of locally produced AngII, which has been well characterized in a variety of tissues and cell types (Reid et al., 2011; Angeli et al., 2013; Lu et al., 2013). Recently, candesartan was shown to be proangiogenic in cerebral microvascular endothelial cells via activation of the angiotensin II type 2 (AT2) receptor (Alhusban et al., 2013). This effect occurred even in the absence of exogenous AngII. These two observations highlight the importance of investigating the potential direct effects of ARBs on tumor cell function and angiogenesis in the absence of exogenous AngII to gain better understanding of physiologic responses. In the current study, the focus was to systematically investigate the effect of clinically relevant concentrations of ARBs on the progression of prostate cancer both in vivo and in vitro. In addition, we investigated the effect of ARBs on tumor angiogenesis and vascular normalization, and the molecular mechanisms leading to ARB action on prostate tumor cells and tumor vasculature.
Materials and Methods
In Vivo Nude Mouse Tumor Xenograft Model.
All animal procedures listed in this article were performed as per the protocol approved by the Institutional Animal Care and Use Committee at the Charlie Norwood Veterans Affairs Medical Center (Augusta, GA; protocol #12-06-049). PC3 cells were grown to confluence in 250-ml flasks. Next, cells were suspended in sterile saline to a concentration of 5 × 106/ml. Cell suspension (100 µl/mouse) was injected subcutaneously in 8-week-old male nude mice (athymic nude mice; Harlan, Indianapolis, IN) (Kochuparambil et al., 2011). Mice were divided into two groups. The groups were subjected to intraperitoneal injections of candesartan (CV-11974 [see Alhusban et al., 2013] dissolved in 100 µl of 0.9% saline) at a dose of 6.5 mg/kg body weight every 24 hours for 18 days (treatment started 6 days after subcutaneous tumor injection when tumors of equal size were clearly visible), and the respective controls were injected intraperitoneally with 0.9% saline every 24 hours. CV-11974 is an activated form of candesartan that does not need to be activated by the liver, and hence can be readily used for both in vitro and in vivo experiments. Tumor diameters were measured with digital calipers on days 6, 12, 18, and 24, and the tumor volume in millimeters cubed was calculated by the modified ellipsoidal formula [tumor volume = ½(length × width2)] (Euhus et al., 1986). Mice were sacrificed on day 24 and tumors were dissected, weighed, and snap-frozen for further Western blot and immunohistochemistry analysis.
Reagents, Cell Lines, and Antibodies.
Human metastatic PC3 and DU145 prostate cancer cell lines were obtained from American Type Culture Collection (Manassas, VA) and maintained in Dulbecco's modified Eagle's medium (DMEM) high glucose (Hyclone, Logan, UT) with 10% fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin in 5% CO2 humidified atmosphere at 37°C. Primary antibodies against phosphorylated (p)–protein kinase B (Akt) (S473), total Akt, p-p38 mitogen-activated protein kinase (MAPK), and p–glycogen synthase kinase 3α/β were purchased from Cell Signaling Technology (Boston, MA). Anti–vascular endothelial growth factor (anti-VEGF) antibodies were purchased from Millipore (Billerica, MA). Primary antibodies against β-actin and laminin as well as AT2 receptor agonist (CGP-121141A; see Alhusban et al., 2013) were purchased from Sigma-Aldrich (St. Louis, MO). Primary antibodies against angiotensin II type 1 (AT1) and AT2 receptors were purchased from Abcam (Cambridge, MA). Anti-mouse and anti-rabbit horseradish peroxidase–conjugated secondary antibodies were obtained from Bio-Rad (Hercules, CA). Alexa-Fluor (488) secondary antibody was purchased from Invitrogen (Carlsbad, CA). Candesartan (CV-11974) was gifted to Dr. Fagan's laboratory from AstraZeneca Pharmaceuticals (Wilmington, DE). Clinically relevant concentrations for the treatment of prostate tumor cells in vitro and athymic nude mice in vivo were calculated using the previously published protocol (Reagan-Shaw et al., 2008).
Candesartan Dose Calculations for In Vitro and In Vivo Experiments.
Two important points were considered when calculating the drug concentrations/doses for in vitro and in vivo studies: 1) dose calculations for intraperitoneal administration of candesartan in vivo, as compared with intravenous drug administration in mice, and 2) comparison of doses used in mice with that prescribed in humans. Therapeutic doses for mice as compared with humans were calculated using the following equation: human dose (milligrams per kilogram) = mouse dose × (mouse Km/human Km) (Reagan-Shaw et al., 2008). According to the manufacturer's recommendation and the literature (Gleiter and Morike, 2002), average daily maintenance doses of candesartan for adults who weigh >50 kg are between 4 and 32 mg/day orally, and the bioavailability of candesartan is 15–40%. Since patients who are diagnosed with prostate cancer are elderly, and given the maximum prescribed dose of 32 mg/day orally, expected bioavailability (the fraction of dose that reaches the circulation) of candesartan is 40%, which is calculated as 32 × 40% = 12.8 mg/day for patients weighing 50 kg or more, and a comparable intravenous dose will be 12.8 mg/day for a 50-kg patient = 0.256 mg/kg. To reach this concentration by intraperitoneal administration, candesartan dose was doubled (0.512 mg/kg) considering the loss of drug through incomplete absorption into circulation and the drug removal through the hepatic system entering through mesenteric arteries. By using the mouse formula (aforementioned) based on human doses, dose of mouse (milligrams per kilogram) = dose of human/(Km of mice/Km of human): 0.512 mg/kg/(3/37) = 6.4 mg/kg (∼6.5 mg/kg).
For in vitro experiments, candesartan concentrations ranging from 0.5 to 25 µM are considered therapeutic, and anything above 25 µM was considered supratherapeutic. Comparisons between therapeutic and supratherapeutic concentrations of candesartan were included in in vitro studies.
Western Blot Analysis.
PC3 cells were cultured in DMEM in six-well plates to reach a monolayer and were serum starved. The wells were treated with serum-free DMEM containing candesartan at concentrations of 0.1–25 µM, or AT2 receptor agonist (CGP-121141A at concentrations of 100 nm to 1000 µM), and control cells were grown in DMEM alone. Whole-cell lysates were prepared using lysis buffer [50 mM Tris-HCl, pH 7.4, 1% Triton X-100, 150 mM NaCl, 1 mM EDTA, 2 mM Na3VO4, and 1× complete protease inhibitors (Roche Applied Science, Indianapolis, IN)]. The protein concentration was measured by the DL protein assay (Bio-Rad). Western blot analyses were performed as described previously (Goc et al., 2013).
Reverse-Transcription Polymerase Chain Reaction Analysis of VEGF Expression.
Both PC3 and DU145 cell lines were cultured to reach a monolayer in DMEM in six-well plates and were serum starved. Wells were treated with serum-free DMEM containing candesartan, and control cells were grown in DMEM alone for 24 hours. Total RNA was extracted using TRIzol reagent (Invitrogen) following the manufacturer protocol. One microgram of RNA was used to make cDNA using an iScript cDNA synthesis kit (Bio-Rad) in a total volume of 20 µl for each reaction following manufacturer protocol. GoTaq green master mix 2× (Promega, Madison, WI) was used for polymerase chain reaction (PCR) amplifications in a total volume of 50 µl. Reverse-transcription PCR amplification was done using an Eppendorf thermal cycler (Eppendorf, Hauppauge, NY) using the following temperature and time periods: initial denaturation at 95°C for 3 minutes, followed by 25 PCR cycles using 1) denaturation at 94°C for 30 seconds, 2) primer annealing at 55°C for 30 seconds, and 3) extension at 72°C for 30 seconds and final extension for 72°C for 5 minutes. Two percent agarose gels were used to detect the RNA bands. The primer sequences used were as follow: VEGF forward, 5′-ctacctccaccatgccaagt-3′, and reverse, 5′-gcagtagctgcgctgataga-3′; glyceraldehyde-3-phosphate dehydrogenase forward, 5′-acccagaagactgtggatgg-3′, and reverse, 5′-agtagaggcagggatgatgtt-3′.
Cell Migration Assay.
Cell migration assay (wound healing assay) was performed as described previously (Goc et al., 2012b; Alhusban et al., 2013). Briefly, PC3 cells were grown on 12-well plates to reach confluence (24 hours), and then scratches were made in the cell monolayers using 1-ml pipette tips followed by treatment with different concentrations of candesartan (0.5, 5, 10, 25, and 200 µM) in serum-free DMEM. Control cells were incubated in DMEM alone. Images for scratches were taken at time 0 and 24 hours. The rate of migration was measured using the equation [(1 − T24/T0) × 100], where T24 is the area at the endpoint (24 hours) and T0 is the area at the start time (0 hours).
Transendothelial Migration Assay.
Transendothelial migration of prostate cancer (PC3) cells was measured using electric cell substrate impedance sensing (ECIS) equipment (Goc et al., 2012a) with human dermal microvascular endothelial cells (American Type Culture Collection) plated on 8W10E+ array chips (Applied Biophysics, Troy, NY). In brief, PC3 cells were plated in six-well plates. After 24 hours, the cells were incubated in serum-free DMEM containing candesartan (0.5, 25, and 200 µM) for 24 hours. To avoid the direct effect of candesartan in the conditioned medium on endothelial cell monolayer, medium with candesartan along with control dimethylsulfoxide containing medium was removed 1 hour after treatment and then supplemented with fresh serum-free medium and subsequent incubation for another 23 hours (a total of 24 hours post-treatment). Control cells were incubated in DMEM alone, then conditioned media were collected and live cells were collected from the plate using cell dissociation buffer [20 mM EDTA in phosphate-buffered saline (PBS; pH7.4)] to avoid integrin/receptor loss due to trypsin digestion. We then added either the cells or their conditioned media directly onto endothelial cell monolayers at a density of 5 × 104 cells/well in 50 µl of medium (or 50 µl of conditioned media). Real-time measurements on the transendothelial migration of PC3 cells were recorded by ECIS up to 10 hours.
Cell Proliferation Assay.
Proliferation of the PC3 cell line was determined using the nonradioactive bromodeoxyuridine (BrdU)-based cell proliferation assay (Roche Applied Science) according to the manufacturer's protocol. In brief, PC3 cells were plated in 96-well flat-bottom plates at a density of 5 × 103 cells per well and allowed to grow for 24 hours. Cells were then treated with candesartan (0.5, 5, 10, 25, and 200 µM) for an additional 24 hours in serum-free conditions. Control cells were incubated in DMEM alone. Cells were then subjected to 5-bromo-2-deoxyuridine assay using the BrdU Labeling and Detection Kit III (Roche Diagnostics, Indianapolis, IN) as done previously (Goc et al., 2011, 2012a). BrdU incorporation into the DNA was determined by measuring the absorbance at both 450 and 690 nm on an enzyme-linked immunosorbent assay (ELISA) plate reader.
Cell Viability Assay.
Number of viable cells was assessed indirectly by means of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay using tetrazolium salt conversion into formazan crystals (Al-Azayzih et al., 2012). In brief, PC3 cells were plated in 96-well plates at 5 × 103 cells per well, and allowed to grow for 24 hours. Medium was then replaced with fresh serum-free DMEM containing candesartan (0, 0.5, 5, 10, 25, and 200 µM). After 24-hour treatment, cell viability was measured using the Cell Proliferation Kit I (MTT; Roche Applied Science) according to the manufacturer's protocol and as standardized in the laboratory. The absorbance at 570 nm was measured using an ELISA reader and used to determine relative cell numbers in each well.
Cell Apoptosis Assay.
Cytoplasmic histone–associated DNA fragments were quantified using the Cell Death Detection ELISAPLUS kit (Roche Applied Science) according to the manufacturer's protocol (Al-Azayzih et al., 2012). In brief, PC3 cells were plated in a 96-well plate at a density of 104 cells per well. After 24 hours, the cells were incubated in serum-free DMEM containing candesartan (0.5, 5, 10, 25, and 200 µM) for 24 hours. Control cells were incubated in DMEM alone. Cells were lysed and centrifuged at 200g for 10 minutes, and the collected supernatants were subjected to ELISA. The absorbance was measured at 405 nm (reference wavelength, 492 nm).
Immunofluorescence staining of the tumor sections for laminin (blood vessels) was performed according to the standard protocol (Goc et al., 2012a). In brief, formalin-fixed, frozen prostate (PC3) xenograft tumor sections from nude mice were subjected to the standard xylene-ethanol dehydration process and permeabilized with 0.3% Triton X-100 in 1× PBS. The nonspecific staining was blocked with 5% goat serum for 1 hour at room temperature. The dehydrated, permeabilized, and blocked tissue sections were incubated with primary antibodies against laminin (dilution 1:750) overnight at 4°C followed by washing with 1× PBS (four times for 15 minutes each). Next, anti-mouse Alexa Fluor 488–labeled secondary antibodies (1:500) were applied for 1 hour at room temperature and washed four times for 15 minutes with 1× PBS. The slides were mounted with Vectashield (Vector Laboratories, Burlingame, CA), and the images were taken by a Zeiss fluorescent microscope (Zeiss Axiovert100M; Carl Zeiss, Jena, Germany). Analysis of the vascular lumen area and wall thickness was determined using NIH ImageJ software.
Statistical Analysis.
All data are presented as the mean ± S.D. of three to four independent experiments. To determine significant differences between treatment and control values, we used Student's two-tailed t test. One-way analysis of variance was used for all of the concentration-dependent analyses in vitro. Significance was set at 0.05 levels (marked with symbols wherever data are statistically significant).
Clinically Relevant Dose of Candesartan Inhibited Growth of Prostate Tumor Xenograft in Athymic Nude Mice.
Our data indicated that candesartan inhibited prostate cancer (PC3) tumor xenograft progression as detected by the size and weight of the tumors after treatment with the clinically relevant dose of candesartan (Fig. 1A). Although we observed a trend in inhibitory effect of candesartan on the growth of tumor xenograft starting from as early as 6 days post-treatment (12 days after PC3 cell administration), a significant antitumor effect was revealed from day 12 postcandesartan treatment (18 days after PC3 cell administration) (Fig. 1B). Additionally, candesartan significantly reduced the weight of prostate tumor xenografts measured after tumor xenograft extraction on day 24 (Fig. 1C). Furthermore, treatment with candesartan resulted in a significant reduction in the number of blood vessels approaching the tumor, suggesting an impairment of PC3 cell ability to attract host vasculature toward the tumor xenograft (Fig. 1D).
Candesartan Had a Modest Effect on the Migration and Apoptosis of PC3 Cells.
Since a clinical dose of candesartan was effective in inhibiting PC3 tumor growth and tumor neovascularization in vivo, we sought to determine if relevant concentrations of candesartan had any effect on PC3 tumor cell function in vitro. Our data showed that relevant concentrations of candesartan used in the in vivo tumor experiment induced a dose-dependent antiapoptotic effect in PC3 cells in vitro, which was observed even at supratherapeutic doses (Fig. 2A). Although modest, the effect of candesartan on tumor cell survival was significant and was in contrast with our data from in vivo analysis of tumor xenografts, which suggested that the effect of candesartan in vivo is much more complicated than its direct effect on tumor cells in vitro. Interestingly, although clinically relevant concentrations of candesartan did not elicit any effect on PC3 cell migration, supratherapeutic doses of candesartan had a paradoxical inhibitory effect, as reported by many other laboratories, which might be due to its toxic effect at high concentrations (Fig. 2B). Surprisingly, clinically relevant concentrations of candesartan did not have any effect on the proliferation or viability of PC3 cells in the short term (Fig. 2, C and D) or in the long term (72 hours) (Supplemental Fig. 1).
Candesartan Did Not Affect the Major Survival Pathways in PC3 Cells.
In contrast to our in vivo data, in vitro data demonstrated a minimal effect of candesartan on the progression of cancer cells. To resolve this discrepancy, we analyzed the major survival pathways in PC3 cells in response to candesartan treatment. Our data demonstrated that therapeutic concentrations of candesartan did not alter the activity of either Akt or glycogen synthase kinase as determined by the changes in activity modulating phosphorylation (Fig. 3A). Consistent with previous reports (Uemura et al., 2003, 2005b, 2006), candesartan inhibited p38 MAPK signaling in a concentration-dependent manner (Fig. 3B), which might be responsible for the modest but significant decrease in prostate cancer cell apoptosis in vitro. A recent study indicated modulation of AT1 and AT2 receptor expression by candesartan in endothelial cells (Alhusban et al., 2013). Hence, we determined if treatment with candesartan will modulate AT1 and AT2 receptor expression in prostate cancer cells. Our study indicated that treatment with candesartan resulted in a significant and concentration-dependent decrease in AT1 receptor expression and increase in AT2 receptor expression in PC3 cells (Fig. 3, C and D, respectively), but not in DU145 cells (Supplemental Fig. 2, B and C), thus suggesting that effects of candesartan on AT1 and AT2 receptor expression, and probably the entire effect of candesartan on PC3 cells in vitro and tumor xenograft in vivo, may be purely limited to PC3 cells.
Candesartan Inhibited Cancer-Induced Angiogenesis in Prostate Cancer Xenograft.
Since candesartan did not elicit a significant inhibitory effect on PC3 and DU145 prostate cancer cells in vitro, we postulated that the effect of candesartan in vivo could be a result of its ability to interfere with the tumor and stromal cell interactions. To assess the involvement of angiogenesis in candesartan-mediated inhibition of tumor growth in vivo, we determined the effect of a clinically relevant dose of candesartan on the expression of the proangiogenic growth factor, VEGF, and on tumor angiogenesis. Accordingly, PC3 tumor xenograft sections were stained with laminin, a marker of vascular density in xenografts, and tumor lysates were subjected to Western analysis for VEGF expression. Candesartan significantly inhibited tumor angiogenesis in PC3 tumor xenografts (Fig. 4, A and B), and this was corroborated with a significant inhibition of VEGF expression (Fig. 4, C and D). Furthermore, our in vitro analysis indicated that treatment with clinically relevant concentrations of candesartan inhibit VEGF expression in both PC3 cells (Fig. 4E) and DU145 cells (Supplemental Fig. 2A) in a concentration-dependent manner. Together, these data indicated that the effect of candesartan on tumor cell VEGF expression, tumor angiogenesis, and growth is indeed a global effect, not cell-specific, and may be via an AT1/AT2 receptor–independent mechanism.
Candesartan Inhibits VEGF mRNA Expression in Prostate Cancer Cells.
Next, we determined whether candesartan inhibits VEGF expression at the mRNA level or if its effect is limited to protein translation. Our study revealed that, in both PC3 and DU145 cells, candesartan treatment resulted in a significant reduction in VEGF mRNA levels at both the 0.5 and 25 µM candesartan concentrations (Fig. 5, A–C), thus indicating that the effect of candesartan on inhibition of VEGF expression in prostate cancer cells is at the transcriptional level.
Candesartan Induces Vascular Normalization in Tumor Xenografts In Vivo.
Since the effect of candesartan in vivo was predominantly on the tumor vasculature, we analyzed the tumor blood vessels to determine the effect of candesartan on tumor vascular normalization, an important feature that determines the efficacy of antiangiogenic and combinational therapies. According to our data, candesartan treatment resulted in significant reduction in the tumor vascular lumen perimeter (Fig. 6A) and lumen area (Fig. 6B) associated with a significant reduction in AT1 receptor (Fig. 6D) and increase in AT2 receptor (Fig. 6E) expression levels, indicating an overall vasodilatory effect of candesartan on tumor blood vessels. In addition, a significant increase in vascular wall thickness in candesartan-treated tumor blood vessels compared with saline-treated controls (Fig. 6C) suggested the ability of candesartan to enhance endothelial-barrier integrity and activation in vivo. Together, these data indicated the effect of candesartan on tumor vascular normalization, potentially via inhibition of tumor vascular permeability, an important feature essential to promote neovascularization in tumors.
Conditioned Media from Candesartan-Treated PC3 Cells Did Not Compromise Human Microvascular Endothelial-Barrier Function.
The effect of candesartan on tumor vascular normalization prompted us to determine its effect on inhibition of vascular permeability by simulating the conditions in vitro using the ECIS technology. Our data showed that conditioned media collected from candesartan-treated PC3 cells preserved endothelial-barrier function in a concentration-dependent manner (Fig. 7A). This finding suggests that candesartan reduces the expression and release of mitogenic mediators such as VEGF by the prostate cancer cells, and that these mediators primarily produce a paracrine rather than an autocrine effect. On the other hand, candesartan-treated PC3 cells did not have a consistent effect on the endothelial-barrier function (Fig. 7B). PC3 cells treated with low concentrations of candesartan induced a trivial breakdown in endothelial-barrier function that was rapidly restored. Interestingly, PC3 cells treated with higher concentrations of candesartan failed to disrupt barrier function of endothelial cells.
Effect of Candesartan on VEGF Expression by Tumor Cells Is Independent of AT2 Receptor Activation.
Finally, we determined whether the candesartan-mediated effect on VEGF expression by PC3 and DU145 cells was through activation of the AT2 receptor. To do this, we treated PC3 and DU145 cells with various concentrations of the AT2 receptor agonist CGP-121141A. Our data indicated that AT2 receptor activation had no effect on VEGF expression in either PC3 or DU145 cells (Supplemental Fig. 2).
Our data demonstrate the ability of clinically relevant doses of candesartan to inhibit prostate cancer xenograft growth. Interestingly, candesartan-induced antitumorigenic effect was mediated indirectly through inhibition of cancer-induced angiogenesis. This finding was confirmed by the lack of significant effect of candesartan on the behavior of prostate cancer cells in vitro. Additionally, whereas conditioned media from candesartan-treated cells were able to maintain endothelial-barrier function in vitro, treatment with candesartan induced vascular normalization in tumor vasculature through inhibition of vascular permeability, thus impairing neovascularization and tumor perfusion.
Candesartan is an AT1 receptor antagonist and is widely prescribed for the management of hypertension and other cardiovascular disorders. It is extremely well tolerated, as are all of the other ARBs), and has an additional advantage of being dosed orally once daily. The pleiotropic properties of the ARBs, beyond blood pressure lowering, include prevention of pathologic remodeling after myocardial infarction, vascular protection, and the reduction of inflammation. The effects of ARBs on angiogenesis are debated and are likely to be dose-, tissue-, and context-dependent (Willis et al., 2011). Candesartan and other AT1 blockers have been demonstrated to inhibit the proliferation of cancer cells when used alone or in combination with other antineoplastic agents (Fujimoto et al., 2001; Miyajima et al., 2002; Kosugi et al., 2006; Kosaka et al., 2007; Chen et al., 2013). This antiproliferative effect was observed with high doses of AT1 blockers that are not clinically relevant and may have serious side effects, such as hypotension and kidney failure. Recently published meta-analyses had confusing results on the risk of cancer among AT1 blocker–treated patients (Sipahi et al., 2010; Mearns, 2011). Accordingly, we assessed the effect of clinically relevant doses of candesartan on the progression of prostate cancer using a xenograft model in nude mice. Similar to previously published data on the use of higher doses of candesartan (Uemura et al., 2006, 2008; Kosaka et al., 2007), our data also demonstrated the ability of candesartan to inhibit the progression of prostate cancer in vivo. This finding has a highly significant translational impact as it provides the first experimental evidence on the effect of candesartan on prostate tumor growth at doses comparable with that used in clinical practice, and can help to resolve the recently raised concerns about the safety of ARBs with regard to the risk of prostate cancer.
The antiproliferative effects of ARBs have been extensively reviewed (Uemura et al., 2006). A common theme among the majority of published literature is the concomitant treatment of prostate cancer cells with AngII (Uemura et al., 2003, 2005b, 2006). In this context, the observed antiproliferative effects of ARBs are basically a reflection of AT1-mediated signaling in cancer cells. This approach does not take into account the possible role of locally produced AngII, which has been demonstrated to be of major importance in other cell types (Reid et al., 2011; Angeli et al., 2013; Lu et al., 2013). In addition, this approach also ignores the possibility of ARB-induced AngII-independent effects which have been demonstrated in other cell types (Alhusban et al., 2013). Another source of complexity in understanding ARB-induced antiproliferative effects is the complex nature of the cancer microenvironment, which has been shown to play a major role in the progression of prostate cancer (Uemura et al., 2005a). To account for all of these variables, we were interested in assessing whether ARB-induced antiproliferative effects are due to the direct effects of the drug on prostate cancer cells. Accordingly, direct cytotoxicity of candesartan was assessed by treating PC3 cells with different concentrations of candesartan in the absence of exogenous AngII. Apoptosis, proliferation, viability, and migration of PC3 cells were assayed. Interestingly, the antiproliferative effect of candesartan was found to be unrelated to direct cytotoxicity against cancer cells. This finding was suggested by the relative lack of effect when candesartan was directly applied to prostate cancer cells. To our surprise, candesartan had an antiapoptotic effect on PC3 cells, and this was observed at all concentrations tested, including the supratherapeutic range. Candesartan-induced antiapoptotic effect was not associated with an increase in either migration or proliferation of PC3 cells. In contrast, higher concentrations of candesartan inhibited the rate of PC3 cell migration. Taken collectively, these data suggest that clinical doses of candesartan do not have an appreciable direct effect on PC3 cells. This finding supports previously published data on the inability of candesartan to affect the progression of prostate cancer cells in vitro (Matsuyama et al., 2010).
Interestingly, candesartan inhibited the phosphorylation of p38 MAPK in a concentration-dependent manner, and at all concentrations used. This finding reproduces previously published data on the effect of supratherapeutic doses of ARBs on prostate cancer in the presence of exogenously added AngII (Uemura et al., 2003). This similarity supports the hypothesis that observed ARB-induced antiproliferative effects are mediated by antagonizing the effects of locally produced AngII. AngII is expected to mediate its proangiogenic effects through AT2 receptors, as has recently been shown in our studies in brain endothelial cells (Alhusban et al., 2013). However, although candesartan treatment resulted in significantly decreased AT1 and increased AT2 receptor expression in PC3 cells and tumor xenografts, this effect was not observed in DU145 cells, suggesting that the common candesartan effects seen may be due to its direct inhibitory effects on AT1, as reported in melanoma (Akhavan et al., 2011). An alternative explanation is the possible AngII-independent effects of ARBs, which require more detailed, in-depth analyses that go beyond the scope of this investigation.
Cancer progression is a well orchestrated interplay between cancer cells and their microenvironment (Uemura et al., 2005a). This interplay can be best demonstrated by cancer-induced angiogenesis (Hanahan and Folkman, 1996). Cancer cells have high metabolic rates and require higher rates of nutrient delivery. To match their metabolic requirements, cancer cells induce angiogenesis, vascular permeability, and perfusion through the release of a variety of angiogenic mediators (Weidner et al., 1993). The lack of concordance between in vivo and in vitro effects of candesartan suggested possible involvement of other cell types in the observed antiproliferative effect of candesartan in vivo. VEGF plays a major role in prostate cancer–induced angiogenesis (Feng et al., 2008; Mahabeleshwar et al., 2008). Our in vivo data demonstrated elevated levels of VEGF in prostate tumor xenografts from saline-treated animals, and a significant inhibition of VEGF expression with a therapeutic dose of candesartan. The effect of candesartan on VEGF expression was also observed in both PC3 cells and DU145 cells. Interestingly, the AT2 agonist (CGP-121141A) had no significant effect on VEGF expression in either PC3 or DU145 cells, thus ruling out the role of AT2 receptor stimulation in candesartan-mediated inhibition of VEGF expression. Our previous studies in a stroke model indicated increased VEGF expression by candesartan (Guan et al., 2011), probably through endothelial cells (Alhusban et al., 2013; Soliman et al., 2014); in the current model, where angiogenesis is mainly driven by the tumor cells, the mechanism leading to inhibition of VEGF expression is not via AT2 stimulation. As reviewed previously, the effects of ARBs on angiogenesis are dose-, tissue-, and context-dependent (Willis et al., 2011)
Since no significant changes in survival pathways and tumor cell function with the clinical concentrations of candesartan were observed in the in vitro experiments, we wanted to rule out that the decrease in VEGF expression in candesartan-treated tumors is due to the paracrine effect from prostate cancer cells. To do this, and to confirm the antiangiogenic effect of candesartan, ECIS technique was used. In this system, endothelial cells cultured to confluence were challenged by either candesartan-treated PC3 cells or their conditioned media, and changes in electrical impedance were measured as a function of time to assess the changes in the endothelial-barrier resistance. ECIS findings demonstrated a protective effect of conditioned media from candesartan-treated PC3 cells on endothelial-barrier function compared with untreated control medium. This finding further supports the paracrine over the autocrine effect of VEGF in prostate cancer proliferation. In support of this, our immunohistochemistry analysis of tumor sections revealed that candesartan induced a vascular normalization effect on tumor vasculature as evidenced by reduced lumen size, increased vessel wall thickness, and more rounded appearance as compared with saline-treated controls. These characteristic features of tumor vasculature are associated with inhibition of tumor vascular permeability and tumor perfusion, thus inhibiting neovascularization and depriving the tumor cells of necessary nutrients to grow, rather than a direct antiproliferative effect of candesartan on tumor cells.
In conclusion, our data demonstrate the ability of candesartan to inhibit progression of prostate cancer through ablation of cancer-induced angiogenesis via inhibition of VEGF expression by prostate cancer cells. Although a role for AT1 and AT2 in candesartan-mediated inhibition of VEGF expression and/or prostate tumor growth is ruled out, existence of an AT2-independent pathway by AngII in candesartan-mediated inhibition of prostate tumor growth and neovascularization needs to be investigated. Nevertheless, our study conclusively demonstrated that clinically relevant doses of candesartan, alone or in combination with other chemotherapeutic drugs, can be developed into a potential therapeutic strategy for prostate cancer. Although monotherapy with candesartan induces vascular normalization and inhibits tumor angiogenesis in prostate tumors in our studies, it will be interesting to know the effect of candesartan in combination with other drugs on prostate tumors. Since vascular normalization is known to affect drug delivery in addition to its effects on tumor angiogenesis, the result of combining candesartan with other drugs can be interpreted in both ways. Additional research will be necessary to address this discrepancy.
Authorship Contributions
Participated in research design: Alhusban, Al-Azayzih, Fagan, Somanath.
Conducted experiments: Alhusban, Al-Azayzih, Goc, Gao.
Performed data analysis: Alhusban, Al-Azayzih, Goc, Gao, Fagan, Somanath.
Wrote or contributed to the writing of the manuscript: Alhusban, Al-Azayzih, Fagan, Somanath.
- Received May 9, 2014.
- Accepted June 30, 2014.
A.A. and A.A.-A. contributed equally to this work.
Funds were provided by the University of Georgia, Wilson Pharmacy Foundation, and in part by the National Institutes of Health National Heart, Lung, and Blood Institute [Grant R01-HL103952] (to P.R.S.). A.A. and A.A.-A. were supported by predoctoral fellowships from Jordan University of Science and Technology. This material is the result of work supported with resources and the use of facilities at the Charlie Norwood VA Medical Center, Augusta, GA. The funders had no role in the study design, data collection, analysis, and decision to publish. Preparation of the manuscript and the contents do not represent the views of the Department of Veterans Affairs or the US Government.
↵This article has supplemental material available at jpet.aspetjournals.org.
- Akt
- protein kinase B
- angiotensin II type 1 receptor blocker
- AngII
- angiotensin II
- angiotensin II type 1 receptor
- angiotensin II type 2 receptor
- BrdU
- bromodeoxyuridine
- Dulbecco's modified Eagle's medium
- electric cell substrate impedance sensing
- enzyme-linked immunosorbent assay
- mitogen-activated protein kinase
- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- phosphate-buffered saline
- polymerase chain reaction
- vascular endothelial growth factor
- U.S. Government work not protected by U.S. copyright


text:
DJ Rocko – 'Ground Zero'
By Jack Barnes
DJ Rocko releases his debut mixtape titled "Ground Zero", featuring Fame Shakur, Hot Boy Tay, Dee Jackson, Herb Osirus, and more; with exclusive production from Deezy On Da Beat, Cruz Davinci, Will A Fool, and Sinista On Da Traxxx. The project is executive produced by fellow Astro Gang member, LaTre'. Enjoy this audio stream below after the jump.
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A photographer friend and I were working our shift at Artomatic Frederick and our discussion turned to selling prints. As one who does not sell enough to begin to think of paying for the paper used, the conversation quickly turned to what prints sell for big bucks and lamenting our inability to participate in this category.
I whined about the fact that Steichen only barely makes it into the top ten while Weston only barely makes it into the top twenty-five (no Adams, no Stieglitz, etc.). I mentioned Rhein II, which sold for over four million dollars, and he told me that any insignificant smudge on a wall could serve for such an image and pointed to one. As he prepared his camera to take a picture of such I one-upped him by using my cell phone to photograph the subject (after which he one-upped me by taking the picture with a much older cell phone).
So for your viewing pleasure I present Untitled, my million dollar picture (which would not come close to being in the top twenty-five). I may print this out and offer it, hoping that someone would make the purchase for FOMO (act now and I will sell it for just a hundred thousand dollars).


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Courtesy of: The American Red Cross
As the season for vacations, barbecues, and town fairs heats up, folks are in desperate need to cool down. While summer provides people with the wonderful opportunity for various outside activities, we need to still be mindful of the summer heat. According to Cindy Erickson, Regional CEO of the American Red Cross, "Excessive heat can be deadly; it has caused more deaths in recent years than all other weather events." In order to beat the heat, these precautionary measures are suggested to have a fun and safe summer.
How to Prepare
- Listen to local weather forecasts and stay aware of upcoming temperature changes.
- Discuss heat safety precautions with members of your household. Have a plan for wherever you spend time— home, work and school—and prepare for the possibility of power outages.
- Check the contents of your emergency preparedness kit in case a power outage occurs.
- If you do not have air conditioning, choose places you could go for relief from the heat during the warmest part of the day (schools, libraries, theaters, malls).
- Know those in your neighborhood who are elderly, young, sick or overweight. They are more likely to become victims of excessive heat and may need help.
- Get trained in first aid to learn how to treat heat-related emergencies.
- Ensure that your animals' needs for water and shade are met.
What to do During Heat Wave
- Never leave children or pets alone in enclosed vehicles.
- Stay hydrated by drinking plenty of fluids even if you do not feel thirsty.
- Avoid drinks with caffeine or alcohol.
- Eat small meals and eat more often.
- Avoid extreme temperature changes.
- Wear loose-fitting, lightweight, light-colored clothing. Avoid dark colors because they absorb the sun's rays.
- Slow down, stay indoors and avoid strenuous exercise during the hottest part of the day.
- Postpone outdoor games and activities.
- Use a buddy system when working in excessive heat.
- Take frequent breaks if you must work outdoors.
- Check on family, friends and neighbors who do not have air conditioning, who spend much of their time alone or who are more likely to be affected by the heat.
- Check on your animals frequently to ensure that they are not suffering from the heat.
Recognize and Care for Heat-Related Emergencies
Heat Cramps
- Muscular pains and spasms that usually occur in the legs or abdomen caused by exposure to high heat and humidity and loss of fluids and electrolytes. Heat cramps are often an early sign that the body is having trouble with the heat.
Heat Exhaustion
- Typically involves the loss of body fluids through heavy sweating during strenuous exercise or physical labor in high heat and humidity.
- Signs of heat exhaustion include cool, moist, pale or flushed skin; heavy
- sweating; headache; nausea; dizziness; weakness; and exhaustion.
- Move the person to a cooler place.
- Remove or loosen tight clothing and apply cool, wet cloths or towels to the
- skin. Fan the person. If the person is conscious, give small amounts of cool
- water to drink. Make sure the person drinks slowly. Watch for changes in
- condition.
- If the person refuses water, vomits or begins to lose consciousness, call
- 9-1-1 or the local emergency number.
Heat Stroke (also known as sunstroke)
- A life-threatening condition in which a person's temperature control system stops working and the body is unable to cool itself.
- Signs of heat stroke include hot, red skin which may be dry or moist; changes in consciousness; vomiting; and high body temperature.
- Heat stroke is life-threatening. Call 9-1-1 or the local emergency number
- immediately.
- Move the person to a cooler place.
- Quickly cool the person's body by giving care as you would for heat exhaustion.
- If needed, continue rapid cooling by applying ice or cold packs wrapped in a cloth to the wrists, ankles, groin, neck and armpits.
Know the Difference
- Excessive Heat Watch— Conditions are favorable for an excessive heat event to meet or exceed local Excessive Heat Warning criteria in the next 24 to 72 hours.
- Excessive Heat Warning— Heat Index values are forecast to meet or exceed locally defined warning criteria for at least 2 days (daytime highs=105-110° Fahrenheit).
- Heat Advisory— Heat Index values are forecast to meet locally defined advisory criteria for 1 to 2 days (daytime highs=100-105° Fahrenheit).
Sun Protection
Limit exposure to direct sunlight between 10 a.m. and 4 p.m., and wear a broad-spectrum sunscreen with a protection factor of at least 30. Reapply sunscreen often. Remember to drink plenty of water regularly, even if not thirsty. Avoid drinks with alcohol or caffeine in them. Protect the eyes by wearing sunglasses that will absorb 100 percent of UV sunlight. Protect the feet – the sand can burn them and glass and other sharp objects can cut them.
During hot weather, watch for signs of heat stroke—hot, red skin; changes in consciousness; rapid, weak pulse; rapid, shallow breathing. If it's suspected someone is suffering from heat stroke:
- Call 9-1-1 and move the person to a cooler place.
- Quickly cool the body by applying cool, wet cloths or towels to the skin (or misting it with water) and fanning the person.
- Watch for signs of breathing problems and make sure the airway is clear. Keep the person lying down.
Download the First Aid App
Another thing people can do is download the free Red Cross first aid app, which puts expert advice for everyday emergencies at someone's fingertips. It offers a great deal of information on preparing for the heat, as well as dealing with heat emergencies. The app is available for direct download from the Apple or Google Play for Android app stores.


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Production Blog
Here's Hoping We Overestimate Another Invasive Pest
ROCKVILLE, Md. (DTN) -- Soybean rust was a best case scenario for an invasive pest. When the tropical pathogen swept up into American soybean fields in the fall of 2004, the country's agricultural community was armed and ready.
By the time springtime rolled around, 35 states had prepared more than 300 "Sentinel" soybean plots, to be checked weekly for the insidious yellow rust spores. A website was launched and ready to track the spread of the disease, which had been known to cause up to 90% soybean yield loss in regions like Africa and South America.
Perhaps most importantly, the university Extension system -- an essential but diminishing institution -- was re-charged and energized. Over the years, funding poured in from USDA, the Risk Management Agency and the United Soybean Board.
The details of this reaction can be found in a recent DTN story, Rust Retrospective (http://bit.ly/…). As I penned it, I found myself hoping I write a similar story 15 years from now -- on a completely different beast.
Old World Bollworm, known as Helicoverpa armigera, isn't well known to American farmers just yet. The pest has long been an agricultural pest of immense influence in other regions of the world -- Australia, Europe, China, Africa and Asia -- but only recently did it breach the Western Hemisphere.
In 2013, it was found ravaging the soybean fields of Mato Grasso, Brazil. A pest of impressive migratory abilities, the moth has been moving steadily north ever since. In September 2014, it was found on pea plants in Puerto Rico. By July of 2015, a handful of male moths had fluttered into the continental U.S. and were found in Florida in the western coastal county of Manatee.
For now, that's as far as the Old World Bollworm has ventured (or rather, been found) in the U.S. But given its broad appetite for over 180 species of plants, including cotton, corn, soybeans, small grains, fruits, vegetables and more, entomologists have told DTN again and again that its spread is likely imminent.
Perhaps most concerning is the pest's ability to develop resistance to insecticides rapidly. Old World Bollworm has already overcome pyrethroids in many parts of the world. Now research has shown that it often carries genes that grant resistance to Bt traits the pest has never even encountered.
Take Vip3a, Syngenta's highly effective Viptera trait targeting aboveground pests like the Old World Bollworm.
The trait has not yet been commercialized in Australia. But scientists from Spain and Australia just published a study showing that "resistance alleles" to Vip3a "can readily be detected in Australian field populations despite no obvious selection."
"Natural variation exists in insect populations that could drive resistance once crops expressing Vip3Aa are introduced unless appropriate resistance management measures are implemented," the scientists wrote.
Vip3a is used in the U.S. -- as are a number of other aboveground Bt proteins, which are deployed broadly each season across both corn and cotton fields. This exposure could give the Old World Bollworm a much greater opportunity to become a Bt-resistant threat, the Australian and Spanish scientists concluded.
"With the recent incursion of H. armigera into the New World, there is enormous potential selection for resistance primarily due to the large areas of corn expressing Vip3Aa proteins to control the closely related Helicoverpa zea and other lepidopteran corn pests," they wrote.
Another study, published in March 2015 by a group of international scientists familiar with Old World Bollworm estimated that $843 million worth of U.S. crops would be vulnerable to the pest, given its climactic preferences.
You can find both studies here: http://bit.ly/…and here: http://bit.ly/….
Soybean rust's threat to U.S. soybean growers never quite materialized. Though it remains a disease of prominence in the South, it has never made it to the Midwest early enough to impact yields there.
But in the wake of the disease's invasion, we were left with a blueprint for detecting and monitoring an invasive agricultural pest.
Like soybean rust, Old World Bollworm has roused funding from USDA's Animal Plant Health Inspection Service (APHIS). In 2015, APHIS committed $1.2 million toward research and surveying efforts -- a twelve-fold increase from the previous year. In 2016, APHIS funding for these efforts continued but declined to $865,000.
Just as scientists watched for rust spores each year, the National Cooperative Agricultural Pest Survey is actively conducting searches for Old World Bollworm. Last year, state, government and academic employees surveyed fields in 26 states for the pest. (Track their surveying efforts here: http://bit.ly/….)
This may seem like too much money and time spent on a pest that isn't established yet, just as the preparation for soybean rust might seem like overkill in retrospect.
This isn't and that wasn't.
Soybean rust was a practice run for a dangerous, landscape-changing invasive agricultural pest that could alter American farming habits forever. We did really well on that test. Let's hope we do just as well with Old World Bollworm and anything else that follows.
Emily Unglesbee can be reached at firstname.lastname@example.com.
Follow Emily Unglesbee on Twitter @Emily_Unglesbee.
© Copyright 2016 DTN/The Progressive Farmer. All rights reserved.


text: By Barry Frankel In Family Portraits On MauiPosted January 7, 2016 Maui Engagement Session True romance is a rare and beautiful thing, and for Dija and Robert it was love at first sight. A chance encounter at a stop light during the drive home brought these two together. A couple of [...] 1 Read More


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Conveniently packaged at a special price. A red Emus soprano ukulele conveniently packaged with a red bag and offered at a special set price. This set (item EUB-5000) includes: - Emus high-gloss red soprano ukulele with machine heads - Red heavy-duty nylon bag with sturdy zipper and reinforced webbing handl All of our ukuleles are inspected and pre-tuned to your choice of tuning before shipping.


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Make it with
Gibson's Finest 12
083664870387 750 ml
- 1 ½ oz whisky
- 1 oz fresh lemon juice
- ½ oz simple syrup
- 1 dash egg white
Add all ingredients into a cocktail shaker. Shake without ice for about 15 seconds to add texture. Add ice and shake for another 10 seconds to chill. Strain into a glass filled with ice and garnish with a lemon twist.
Why use egg white?
Adding egg white to a cocktail isn't about taste, it's all about texture. As you shake the cocktail, the egg white adds a delightful frothy texture that brings a whole new dimension to your drink.
But is it safe?
Absolutely! Using egg white in a cocktail is 100% safe. As with any food item, make sure you wash your hands and the egg shell itself prior to use.


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Welcome to the RF Cafe Jobs website, where a mix
of engineer, technician, manager, and sales job opportunities are posted on behalf
of companies that are actually doing the hiring. If you are responsible for hiring
in your company, please contact me AFTER reading the
Terms of Use. See most recent
list of job openings here.
This list provides a good sampling of what
is available if you are seeking to enhance your career, whether it be via searching
for a new job, improving your status and abilities in your current job, or starting
your own company.
Engineering Jobs listed on RF Cafe.
Job Resources: 2014 |
2015 |
2016 |
2017 |
2018 |
May 6, 2019 - Advanced Test Equipment Rentals is a leading provider of test and measurement equipment. ATEC has established solid relationships with industry leading manufacturers worldwide. ATEC offers competitive pay and benefits. Advanced Test Equipment Rentals is looking for a Technical Support Representative. Our ideal candidate has knowledge of electronic testing equipment. Please visit our website at www.atecorp.com to learn more about our company.
What Will You Be Tasked With?
What Are We Looking for in Candidates for This Position?
What Does ATEC Have to Offer You?
About About Advanced Test Equipment Rentals
ATEC is a leading provider of test and measurement equipment, Advanced Test Equipment Rentals has established solid relationships with industry leading manufacturers worldwide. ATEC offers competitive pay and benefits. Please visit our website at www.atecorp.com to learn more about our company.
Jerami MaganaSenior HR Generalist Advanced Test Equipment Corporation10401 Roselle Street San Diego, CA 92121 Email: email@example.com Phone: 858-558-6500 ext. 210 Web: www.atecorp.com
Posted May 5, 2019
RF Cafe began life in 1996 as "RF Tools" in an AOL screen name web space totaling
2 MB. Its primary purpose was to provide me with ready access to commonly needed formulas
and reference material while performing my work as an RF system and circuit design engineer.
The Internet was still largely an unknown entity at the time and not much was available
in the form of WYSIWYG
All trademarks, copyrights, patents, and other rights of ownership to images and text
used on the RF Cafe website are hereby acknowledged.
My Hobby Website: AirplanesAndRockets.com


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It sounds simple enough: Hire and promote the best among us. Do what you must to entice high performers who are competent in areas of leadership, strategy, strategic thinking, etc. Find the superstars – the industry rock stars – and get them to lead your teams. And when you can't find or recruit them, do your best to develop them, hope for them and even steal them when necessary to ensure your organization holds a strategic and competitive advantage to achieve success.
Oh, but my clients – and my own management experiences – tell me it's not so simple. Many qualified candidates and experienced managers don't actually know what strategic thinking is or what it looks like. Words like strategy, strategic thinking, strategic thinker and strategic leadership have all become part of the latest array of buzzwords that managers use in executive and team meetings and candidates put on their résumés without truly understanding the nuts and bolts of it all.
Strategic thinking is a highly sought skill. Whether you are the executive trying to put together a strategic leadership team or the applicant on the prowl for your next position or promotion, this is useful information for you. There are several things you can do to show that you are a strategic thinker. You can practice these six habits of true strategic thinkers as well as demonstrate the following three unmistakable signs of strategic thinking.
1. Strategic Thinkers Make Better Decisions By Being More Reflective
Strategic thinkers do it differently. They understand and dig deeper with regards to analyzing processes, developing and applying performance metrics, collecting data and producing analytics to make more informed decisions. Strategic thinkers have high levels of critical thinking skills. When it comes to decision making, they are more contemplative and ask more thoughtful questions like the ones listed below. They challenge themselves, colleagues, employees, supervisors and executives to consider answers for these questions.
Strategic thinkers are more observant and reflective than the average person. Here are some questions they might process through and consider in order to make better decisions for any particular issue, problem, event or strategy.
2. Strategic Thinkers Advance Their Careers By Being Better Leaders
Strategic thinkers are – more often than not – all over leadership. Not all great thinkers are great leaders, but the ones who are leadership material are likely better leaders than those who fail to think strategically. The sum of the whole is much greater than the individual parts.
Strategic leadership is what happens when leaders move beyond the role of having and communicating a vision to fully understanding, thinking about, planning and executing the necessary strategies to realize it. I further define it as what happens when we are exploring, deciphering and interpreting the best concepts for achieving organizational excellence, anticipating and responding to change and aligning organizational systems, processes and people with enhanced services and/or products.
But it's not just about your organization. If you want to advance your brand and your career, be more strategic. When an individual is being 'strategic' about it, she is focused on and considers both the hard and soft side of the system and weighs the implications and consequences of decisions to determine whether or not they impact and affect the team's ability to accomplish strategic outcomes.
If you are doing this – connecting the dots and taking into account the entire system – you are being a strategic thinker who also demonstrates exceptional leadership. These two competencies together are what we know to be strategic leadership.
To learn more specific things that strategic thinkers do to get promoted, read the companion article to this one "3 Things Strategic Thinkers Do To Get Hired In Senior Management Positions."
3. Strategic Thinkers Demonstrate Organizational Value By Creating More Strategic Thinkers
To demonstrate their value to the team and the organization, strategic thinkers ask questions that reveal their propensity for and commitment to strategy. It is important to find and balance the value in strategic thinking. You can't ignore strategy and be a strategic thinker. You can't disregard the importance of delivering value as a leader and be a strategic thinker. And you can't be too busy to make it a priority to develop a culture where others are encouraged to do the same. Strategic thinkers are the ones in the organization who ponder and prompt questions that are designed to create even more strategic thinkers.
Again, the benefits of strategic thinking are twofold. The advantage is not just for achieving organizational goals. Strategic leaders – individuals who demonstrate their competence and talent to both lead and think strategically – have a unique advantage when it comes to owning their careers as well as leading their organizations. If you apply these strategic thinking principles to areas of career advancement, salary negotiations, relationship development, networking, performance agreements and brand development, you will surely experience immense professional benefit.


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- Infantry
- Cavalry
- Artillery
- Safety
- Waterloo
- About Us
- Re-enactors Guide
- The World of King George
- Of All the Kings Men
- Of England and the English
- Of Fashion and Manners
- Of Food and Drink.
- Of Other People, their countries and manners
- Of Pastimes and Politics
- Of People on People and the Nicknames they used.
- Of Words, Phrases and Things from Dictionary
- Of the City our Towns and our Counties
- Of the Professions
- Almanac
The Grenadier Guards
Submitted by Wagg on Mon, 11/01/2010 - 09:09.
By general order on the 29th of July 1815.
Saturday, 29 July 1815
By general order on the 29th of July 1815.
The First Regiment of Footguards are granted the title 'Grenadier'.
This honour was awarded to the regiment for its gallant service at Waterloo.


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Yes, getting too hooked with social media and spending way too much time in front of the computer could make you gain weight. However, the social media could also be a powerful tool to help you in your weight loss journey. Want to know how?
First, you could go searching for fitness bloggers. Set up a Tumblr account and follow people who are having blogs regarding their fitness routines and activities. Tumblr is not only a mere blogging site. It is also really social. You could interact with the people you follow by sending them asks and you could also interact with people who are trying to lose weight just like you. Twitter is pretty much the same. It is just that with Twitter, you do not have to spend about five minutes reading an article. Twitter feature microblogging which means you could get really helpful one-liner fitness tips. Social media and the Internet in general are great tools for feeding your mind with the knowledge you would need.
Social media gives you the opportunity to interact with people who are in the same journey as you or who have succeeded in their weight loss efforts wherever they are in the world. Share gym stories with a newfound friend from France or ask a vegetarian from Thailand on how to make really delicious vegetable-based meals for your diet. Not only does this give you connection to people who may help you. Engaging in social media also helps you feel that you are not alone in this journey.
Also, tweeting or posting your current fitness status or experiences on social networking sites may help you get and remain motivated. It keeps a record of how well you are doing in your endeavors and it shows you a timeline of your progress. It does not have to be a Pulitzer Prize winning piece. Just pour your heart out. After all, it is your Facebook, Twitter, and Tumblr.
Not only could you write articles or even short sentences. You could also post pictures and videos of your workout, diet and many other efforts to lose weight and be fit. Doing so may also help you track your progress, and better, they may help encourage people who are also on their journey especially those who are having a hard time. Get motivation from your friends wherever they are as long as they see a status update or a newly uploaded photo.
However, be reminded that you are not living online. Do not be so hooked that you are only doing your efforts to please people online. Run every morning, eat healthy meals, and hit the gym for your own wellbeing and not to impress anybody. Also, do not forget to connect with real-life people. Interact with those who are physically present. While social media could be a great help in your weight loss journey, nothing beats helping hands and shoulders to lean on that are actually right beside you. At the end of the day, your success or failure is up to you.


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- A Church without Spot of Wrinkles-Eph 5:27
- A Church that is dead to the flesh-Rom. 8:6, Gal. 2:20, 1 Cor. 15:31
- A Church that does not follow the world pattern but Christ pattern-1 John 2:15-17
- A Church that does not compromise her faith for worldly affairs-Luke 9:62
- A Church that base all her teaching and doctrine on the true word of God-Matt 5:37, James 5:12
- A Church that does not harbor Sin but rather expose it-Prov. 28:13, 1 John 3:8
- A Church that speaks the truth at all times-John 8:32, Prov. 12:19, Prov. 23:23
Prepare to Your Meet God: How Prepared are You?
Jesus is coming soon for a Glorious Church
- A Church that shun hypocrisy and doctrine of men-Col. 2:8, 2 John 9,10
- A Church that does not graven any form of image(s) of worship-Exo. 20:4,5,23
- A Church that is ready to die for the sake of the Gospel Phil.1:21


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Bosque de Los Aluxes is one of Playa del Carmen's newest luxury condominium facilities. It has a convenient location on 5th Ave at 34th Street in the popular North End of Playa del Carmen. This is a quieter section of 5th Avenue and yet it's just a 5 minute walk to North Playa del Carmen beach near Shangri-La Caribe, or the cafes, restaurants and shops in the "Little Italy" section of Playa which are just south of Bosque de Los Aluxes.
This Bosque de Los Aluxes building of modern architecture, and pure lines, respecting nature and mingling with the environment through the use of traditional materials thus minimizing the impact. The building was designed with some basic premises in rnind: spacious units, privacy, views of each unit, and a direct connection between balconies and the living room in each unit.
The building is made up of three levels connected in the first floor by the El Callejon de los Suenos, a commericial area, that is actually a continuation of the 5th Avenue. The total built surface area is of 54000 sq.ft, combining the units with very well adapted open areas. The 16 units are organized in three different types: duplex apartments with three or four bedrooms, duplex apartments, with two bedrooms,of about 970 sq.ft, to 2100 sq.ft. plus deck.
El Callejón de los Sueños leads to the general entrance.The building has two circular elevator shafts strategically located in order to connect every unit, wich can be accessed from the parking areas in the first floor. To view the property or for information contact us at www.american-development.com.


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Morning Bulletin
- There was a second consecutive blowout UK Manufacturing Production print Friday morning, which saw Sterling rally in FX space. There was a 2.1% rise in Manufacturing Production for December, smashing the expected 0.5%.
- Further bolstering Sterling, there was a narrowing in the UK's Goods Trade Balance reported. A deficit of 11.5Bn was expected however the actual figure was 10.9Bn.
- Canadian data took centre stage Friday afternoon and it didn't disappoint with 48,300 (-10,100 Exp) more people in employment in January and the official unemployment rate dropping to 6.8% from a previous 6.9%.
- There were also two US economic data releases of note Friday afternoon:
- Import Prices m/m 0.4% as expected
- Prelim UoM Consumer Sentiment 95.7 (97.9 Exp)
- Speaking Friday afternoon ECB member Nowotny said the central bank would look to discuss options regarding their QE program towards the middle of 2017; adding at present there is no reason to deviate from the current plan.
- Japan Prelim GDP q/q 0.2% (0.3% Exp)
- Speaking overnight, US Fed Vice Chair Stanley Fischer acknowledged that there is significant uncertainty about US Fiscal Policy under Trump's administration; however he said the Fed would continue to operate according to the law, maintaining full employment and hitting 2% inflation.
- Speaking on German radio EU President Juncker said the Greek plan is 'on shaky ground'.
- No data releases of any note due today so could be a quiet day in progress.


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Talent Your Body
TALENT YOUR BODY – learn to recognise your resources and generate vital strength!
My professional experience has taught me the great virtues of the body:
- It informs you about your posture, your position in a space or social environment, how you feel, what you see and what you do or don't like, at all times due to its proprioception which means perception of yourself.
- It provides vital support, because if you are strong and feel good about yourself you have the spirit and motivation you need to pursue your goals.
- It shows you who you want to be with, whether a social environment tires or stimulates you, who you find easy to love and who you fight with. This information is indispensable if you want to learn from your social evolution.
- This indicator works when everything's going well and also when everything seems to require an incredible effort. It's a road towards your talent.
- It leads you to the best decisions, for the best state of balance between body and mind.
Being able to see, feel and experience all these qualities, gain a better understanding of
yourself from different perspectives, and leave behind: repetitive patterns from the past, obsolete beliefs, limiting pain and tensions, unexpressed emotions and/or old ideas about yourself, allows you to know and feel who you are today and to write your own future.
People have two main skills that stimulate us and make us feel whole. One is to feel recognized by our social circle, to feel seen and touched. The other is to learn through and from the difficulties we encounter throughout our lives. These skills are with us at all times, irrespective of the nature of our family, cultural environment, age or experience.
TALENT YOUR BODY is a systematic education that covers body and mind, and aligns them with our intentions and current challenges. Everyone is a world, so there is no pre-established programme. The process is focused on maximising your and the group's potential. That means:
Connecting with your body so that you feel your best potential. This will allow you to form connections with others that lead to genuine and real relationships, in which everyone can give their best, socially and culturally. This will allow you to create things together that benefit the whole of the community.