text:
Litchfield National Park Day Tour from Darwin
Read what other Viator.com travelers think about the Litchfield National Park Day Tour from Darwin. What they loved, what they liked and what they think could be improved, it's all here to help you make the most of your next trip.
Click here for tour details, schedule and pricing
Not to much driving. Cold bus - I gut sick after that. Amasing sights!!!
Excellent day trip, You get to see and do a lot in the one day. Recommend it to anyone.
I enjoyed the relaxing manner of this trip. The waterfalls were amazing and very beautiful
Great scenery. Very informative tour driver
Stunning scenery, a must do day trip when in Darwin. Great guide with lots of knowledge.
A great tour with a knowledgeable and fun guide/driver. The rock pool swims were refreshing, although the second pool with the steps was easier to get in and out of than the first one. A nice salad and cold meat lunch, and there was plenty of cold water carried on the bus, so we could top up our water bottles whenever we wanted. Definitely worth the money!
Swimming amongst fishes is quite interesting .
A very pleasant day. Swimming in the waterholes was great but very dangerous on slippery rocks to get in.
it's ok to do, only because in darwin there's nothing else to do! so you visit the parks, if you are lucky you see some crocodiles, its very hot, many many flies around, food is not good, you are longer in the bus than outside... this is a kind of holidays..!
Lesley-Ann M
It was a great day, absolutely jam packed. We were well looked after. I would recommend it to a mature person as I was the youngest person on the trip.
Jacqueline M
United Kingdom
A long day with a lot of travelling. Some older people on our trip found the walks to the various falls very arduous in the heat, so be aware of this.
Paul S
The highlight are the Billabongs and rock pools we stopped at. Come prepared for swimming! The trip to/from Darwin is long and less rewarding, so also come prepared to snooze.
Not as amazing as I expected.
It is a "small" tour. Lots of time to relax. It is possible to hike on your own. But don't expect a decent lunch, bring your own food.
We all enjoyed the trip to Litchfield well worth a visit, very enjoyable, the driver(Mick) was great very knowlegable. The only downside was we travelled on a small minibus and although not full, there was nowhere to store bags and wasn't that comfortable for a long journey.


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subset: commoncrawl


text:
Perry pushes goal streak to 7 as Ducks top Wild
By Forrest Lee, The Sports Xchange
Minnesota 1 Anaheim 2
ANAHEIM, Calif. -- Corey Perry is wielding a hot stick.
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text: Web Crawling Jobs 65 were found based on your criteria Post a job. It's free! Newest Jobs First Oldest Jobs First Most Employer Hours Least Employer Hours Most Dollars Paid Least Dollars Paid Highest Feedback Score Lowest Feedback Score 1 2 3 4 5 … › » Real Estate Title Examiner County Records Data Mining Scraper Hourly – More than 6 months – 10-30 hrs/week – Posted 1 day, 23 hours ago New Start-up Real Estate Investor in need of a freelancer(s) to do real estate title research data mining/ scraping research of US Counties Real Estate Property Records. The web date mining/ scraping research will require you to perform due diligence research of county real property records of court(s) website(s) which may include title searching and examining, property tax records search, & property owner name search. After collecting the data from the county court(s) website(s), you will be required to place it into an excel file and manually input data onto provided worksheet in preparation for final submission(s). Job Includes: -Request & obtain realty property list from county -Perform current owner search using online USA county websites -Research debts & liens encumbers of title property -Retrieve documents specifically requested within the instructions -Submit completed results via email attachment (with completed spreadsheet, notes, & questions within the email) -Your work will be reviewed for accuracy Must be are able to demonstrate following: -Dependability (complete the assignment on or before the due date & time) -Independence (ability to follow directions & communicate with manager via email and/ or google hangouts only) -Availability (respond to work orders during US hours 10am to 6pm EST) -Detail-Oriented; ability to locate all requested documents, where available. -Proficient in English -Prior Real Estate Title Experience. Very ... administrative-support data-analysis data-entry data-mining data-scraping internet-research lead-generation legal-research microsoft-excel real-estate-appraisal real-estate-idx real-estate-law virtual-assistant web-crawler web-scraping Fetch specific data from a website Fixed-Price – Est. Budget: $20.00 Posted March 27 2015 I need a simple web crawler that'll fetch info from a specific website web-crawler web-scraping Find supplier of dried raspberries for delivery to Australia Hourly – Less than 1 week – Less than 10 hrs/week – Posted March 26 2015 Need good web researcher to find dried raspberries to deliver to Australia. Here are two US suppliers: http://www.greenbulk.com/dried-rasberries-case-p-572.html https://nuts.com/driedfruit/red-raspberries/premium.html Please note : 1. we are not looking for "freeze dried raspberries". 2. product must be premium quality 3. the cost of air freight to Australia is also a key consideration. internet-research lead-generation market-research web-crawler Data extraction from rottentomatoes.com with import.io or similar Fixed-Price – Est. Budget: $50.00 Posted March 26 2015 Hi, I need a spreadsheet that contains the information shown in the attached image. All data can be found on: http://www.rottentomatoes.com/critics/authors I tried it myself with import.io and I almost got there but there are some problems which I am not able to solve with my limited skills. One challenge (at least for me) is to enter the scraped data into existing columns (meaning not creating new columns). For example, say critic 1 rated ... data-mining data-scraping web-crawler Develop a Scrapy Crawler that able to crawl 900 Domains and extract content from a defined div class Fixed-Price – Est. Budget: $85.00 Posted March 25 2015 I need a scrapy crawl that crawl all domains containing in a csv file. The csv file is attached in this job. The crawler should crawl every url on this domains and try to find a specifc div class The div class is: abstractcomponent openinghours here are some example urls with this div-class: http://www.mercedes-benz-berlin.de/content/deutschland/retail-plz1/berlin/de/desktop/passenger-cars/about-us/locations/location.6150.html http://www.europa.mercedes-benz.be/content/belgium/retail-m-r/mercedes-europa/fr/desktop ... python scrapy-framework web-crawler web-scraping xml Finding email addresses from lists of websites Hourly – 1 to 3 months – Less than 10 hrs/week – Posted March 25 2015 Each day I will provide you with several hundred blogs and websites. I want you to go through each link and find the email address. Automated is preferable, manual can be considered too. data-mining data-scraping internet-research lead-generation microsoft-excel spreadsheets web-crawler web-scraping TicketBot Ticket Drop checker Hourly – Less than 1 week – Less than 10 hrs/week – Posted March 24 2015 I'm looking to hire someone with experience in developing TicketBot/Ticket Drop Checker/Crawler to build ticketbot. Please do not bid/apply if you have no experience in ticketbots . web-crawler Web Researcher Needed Hourly – More than 6 months – 10-30 hrs/week – Posted March 24 2015 Hello, I am in need of good web researchers who can browse through the internet and compile lists of relevant websites based on certain topics. Candidate should know how to use excel, not doesn't need advanced knowledge. This is an ongoing need so if the relationship works out we'll have steady work coming your way. Thanks, Ethan data-entry internet-research php web-crawler web-scraping Lead Generation Professionals for startup Hourly – 1 to 3 months – 10-30 hrs/week – Posted March 23 2015 Looking for a lead generation person/team to create social profiles around a database. We will provide the webistes and tools to generate leads for our sales team. b2b-marketing database-management lead-generation linkedin-api outbound-sales sales socialengine web-crawler Develop new data-mining system (c#, .net-framework, oracle) Hourly – 3 to 6 months – 30+ hrs/week – Posted March 23 2015 I need a data-mining system that manages documents inside. The following features are required: - Searching documents over internet by certain criteria like keywords, countries etc. - Search providers (Google, Yahoo, maybe more) should be implemented as plugins and should be able to be added at runtime - Refining search results to have relevant sets - Analysing documents with special rules by tags, keywords. - Checking existing database of documents to have the latest versions - Updating database with new documents automatically or moderating manually Application ... asp.net-mvc c# data-mining .net-framework web-crawler web-scraping


text:
Front mount drawer pulls - how do they hold up?
My pulls are still back ordered. I'm so annoyed. I saw someone post that RH is having another F&F sale this weekend, and I'm about to cancel my order and just go buy the stuff I really love. I think this might be a sign. Lol
My builder discouraged me from the Gilmore pulls because the pulls screw in from the front. He thought that over time the continual force of pulling the handles in the same direction of the screws would cause them to loosen. He thought that eventually the screws would simply not hold up and the pulls would be loose all the time. He said he wouldnt have a problem installing them if that's what I really want, but he did want to warn me.
Has anyone had front mount pulls over a long period of time? Will they drive me nuts in 10, 20, 30 years?
Don't know anything about front-mount pulls, but as far as the screws go, there is a loctite product that will supposedly prevent the screws from loosening up, but the screws can be removed if/when desired (as opposed to their product that is permanent).
I don't see any reason that you cannot use machine screws (i.e., bolts) from the front, rather than wood screws. You will need to do two things for this to work: choose exactly the right length machine screw (with a tolerance of +0, -1/8"), and countersink the space for the nut.
For a normal installation that uses machine screws from the rear, you generally have to countersink for the screw head anyway, so the second item is not a real big deal. Neither is the first. Also, machine screws are available with any head style, so you can match the look of your wood screw.
I'm guessing that the pulls have screws where the heads match the finish of the pulls (I have chrome bathroom hooks that screw from the front that have matching screw heads) so you may not be that free to change the screws to bolts. I think your guy has a point but given the nature of drawer hardware now and assuming your kitchen doesn't get used harshly (ie kids or heavy handed spouse) I'm guessing it could be quite a long time before they would loosen and there are ways tighten stuff up if necessary. What sort of wood are we talking about?
Thanks everyone!
We have maple cabinets with plywood construction. I don't really know/understand much more about them. They're Medallion brand cabinets, painted white. Our hardware will be polished chrome.
Well, I went ahead and cancelled my order. It's so frustrating because I ordered August 19th, and there was no indication that anything was back ordered. I received my knobs but not my pulls. They called about 2 weeks after the order and told me it was back ordered until mid-september. They also failed to send me a couple other things I ordered and claimed to have sent it. Nothing but a headache. Anyway, I guess my lesson is that you get what you pay for. At least I can get a lot of the RH stuff locally and order the rest.
I think it was a sign too. My heart was really set on the RH style, but they were much more expensive and my builder wasn't crazy about them. Well, we just went into contract with our current house and the new house is supposed to be completed in 3 weeks. I NEED my hardware!
It all works out! Happy sigh of relief. :-)
Lazygardens- that's exactly what I wanted to hear! :-)
I ran over to RH this morning, and man, they really don't have much in stock. I walked out with 9 Gilmore pulls, 1 Gilmore knob, and 1 bistro pull. Really? I ordered a zillion others online from the store so I got free shipping. I also fell in love with their lightbulbs they had displayed in their chandeliers. Those can only be purchased online. Is RH just a showroom these days?!?!?
I had pulls for 18 years and they never got loose. As a matter of fact, when we were staging the house to sell, I decided to update the pulls. Had a heck of time getting the old ones off. They were so tight, we broke a few of the screws.
I wouldn't worry about it.


text:
Orchestrating transient cloud instances with traditional configuration management tools usually requires pre-installation of agents, intimate knowledge of often complex syntax and perhaps most importantly, hard-earned time.
What if you could exercise complete control over your instances through a simple language, with the ability to pick any machine image as a building block for your services with no requirement for agents?
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View the on-demand webinar as we look at exactly what makes Ansible the perfect tool for simplifying cloud automation and how Eucalyptus, with its EC2 API compatibility, can help unlock potential hybrid scenarios using common Ansible playbooks.


text: What would you do Wednesday! A friend and fellow church member comes to you with a ministry idea. He recently spent quite a bit of time studying church history and apologetics. Believing God gifted him with discernment, he decides to start an online discernment ministry so he can use this gift. He wants to be known for standing up for and promoting truth! However, you have concerns from knowing him for several years and observing his Facebook interactions. You ask if he is the humility, temperament, and biblical grounds to start such a ministry. You also ask what he will …keep reading »


text:
Starship Pitch (up/down) Limit.
View Single Post
Archived Post
Lt. Commander
Join Date: Dec 2007
Posts: 120
# 14
Of cause there are also the dozen or so DS9 shots showing the Defiant flying upside down, doing loops, flying vertically and "swimming" on its back.
So limiting our ships angles is un-cannon.
Paradise Lost -> Defiant fighting the Lakota, Upside down, and rolling around it self.
Sacrifice of Angels -> Defiant does loops.
For the Uniform -> Defiant pitches diagonally to avoid a colliding pylon.
And thoose are the ones I remember just like that.
In TNG there is also the "relics" episode, where the Enterprise pitches 100% vertically (while flying ahead), to get through the doors of the dysons sphere.
Of cause there is the Enterprise episode "The Expanse", there the NX class does a loop to get behind a BoP to kill it off.
I have no problem with loops or flying upside down. I'm just not fond of nearly straight up/down turning abiilty. You can have one without the other.


text:
"Nice hotel in the airport"
Nice views from the room of the tarmac and surrounding mountains.Genuine Expedia guest review
"Fairmont Vancouver Comfort"
We arrived in the evening ater a ten hour flight. We didn't have to leave the airport to access the Fairmont. Valets were waiting to help with luggage, beds crisp and pressed, comfortable room, good restaurant and spa. That was just what we needed plus a very friendly staff.Genuine Expedia guest review
Pretty darn darn fancy!! Just really $$$$Genuine Hotels.com guest review
Very pleasant, fun to be on the airport property. But frankly, it's ridiculous to charge EXTRA for wi-fi when the room already costs $280 a night. I've stayed in $40 motels that DO NOT charge for wi-fi.Genuine Hotels.com guest review
"Very convenient for early morning trips"
The hotel is connected to the airport making it very convenient for early morning trips, especially international ones. Our room was very quiet. You'd never know you were at the airport. The front desk staff were very friendly. The room was modern, clean and featured everything we needed. We were only there long enough to sleep before a morning flight, […] so we didn't experience many of the other services or features. It is a bit expensive but you're paying for the location / convenience. Read moreGenuine Expedia guest review
"Best place to stay when you have a 6 a.m. flight!"
When I called for a 3 a.m. wake up call; a 4 a,m, wheelchair and porter they were right on time, helped me from my room to the Airline counter and handed me off to another wonderful transporter all the way through to my gate. I stay at this Fairmont every chance I get. There is no better way to start your day off for a flight than a good nights sleep […] at Fairmont Vancouver. Read moreGenuine Expedia guest review
"Thee Best Airport Hotel"
I have just completed by 5th stay at this hotel. It is located directly off of the US flight section of the airport. It is stylish, the staff of competent and friendly, and the rooms are clean, spacious, and quiet (yes, quiet), and the beds are fabulous. The gym and pool are fantastic. If you feel the need to head to downtown Vancouver, it is an easy […] trip on the Canada Line which has a station in the airport. Yes, you can find less expensive off-site airport hotels, but the convenience of this hotel is second-to-none. I will be back. Read moreGenuine Expedia guest review
"Where do I put my water bottle on the treadmills?"
If I have to nitpick then you know the hotel is generally excellent and, of course, you cannot beat the location. Departing on an early flight is easy and the 24-hour gym allows you to workout before that early flight. Do not forget to buy a bottle of water in one of the airport vending machines before entering the hotel. The bottle in the room is too […] large for the slots on the gym machines. My issue is with the design of the treadmills. The slot for the water makes the bottle stick out perpendicular so your fight hand bashes it when running. The slope of the rest of the area in front of the runner makes it impossible to put the bottle any place else. You can always tell when someone who does not workout is involved in selecting equipment. The solution: make sure the water bottle cap is on tight and insert the bottle upside down. It keeps the bottle out of the way of your fist and is the best you can do. Read moreGenuine Hotels.com guest review
"Desk Clerk Saves The Day!"
A great sleep for the weary traveler. Quiet and comfortable. Got to registration about 6:30PM. Clerk was confirming accommodations that included a King bed vs the 2 Queens my 18 year old son and I were expecting. All I did was provide a 'funny look' during that confirmation standing with my son and he with out fuss, flinching or blaming me for the RSVP […] oversight...changed it to a set of Queens and we were off to the room. Superior service. Thank you! Read moreGenuine Expedia guest review
"close to airport"
clean, nice staff, beautiful view from window, nice shampoo, good smell in the roomGenuine Expedia guest review
"under impressed overall."
I would say my stay at the fairmont vancouver airport was ok. I picked this hotel for the convenience of location and it's 4 star rating. I had never stayed at a 4 star hotel. The staff was friendly and helpful. The hotel itself is very beautiful. That being said I did some things that were not as I expected them to be. When I checked in I was upgraded […] to one of ther rooms with a view. I thought wonderful we can see the plans land and take off well this was not the case the side of the hotel we were at was a view of the plains parked to let the passanger of not sure I would call this a view room though the mountains were pretty I guess. The room itself was nice clean and spacious with a very comfy bed. The bathroom was very nice on the eyes with a large bath and separate shower. However a pon taking a bath I saw that the tile surround to the tub was dirty and the next morning that very nice looking shower had terrible water pressure. My husband and I had dinner in the restaurant at the hotel and the food was very good but seemed to take an extra long time between our starter and main course. The waitress was nice but uninformative about the menu options. Over all my stay was Ok but not really worth the price I payed for the room. Read moreGenuine Hotels.com guest review
"Beautiful hotel"
We were celebrating our 30th anniversary! Hotel was just gorgeous but the staff were the ultimate best. My daughter-in-law was sick with morning sickness the first morning and the staff was so helpful in getting us down to the health clinic. Then my husband got intestinal flu on Friday and then me and then my son followed suit! So, we all stayed in […] our room for the majority of the trip except for being taxied to the doctor. The valet driver that brought me back to the hotel from the dr was exceptional! Staff worked hard to clean our room. Would love to stay again! Read moreGenuine Expedia guest review
"Convenient, convenient, convenient!"
The convenience of being smack inside the airport is well worth the expense. The Canada Line makes transportation in & out of airport super easy. The rooms were quiet, despite airplanes taking off & landing. Rooms were immaculate, roomy & comfortable. One room, facing the tarmac, did have slight odor of fuel fume. Maybe that room's HVAC system or seal […] has been compromised. Read moreGenuine Expedia guest review
"Needs work"
The wash sink had several cracks, Some of the walls near garbage cans were not clean, you could see soda stains on the wall paper in the room, no power outlet near the bed ,I crawled behind the night stand to see if I could reach the plug and it looked like it had not been dusted for a few months.Genuine Expedia guest review
W always stay at the hotel on Dec 28th every year a we have an early morning flight back to the desert. We love the hotel and the staff is very friendly.Genuine Expedia guest review
"Oasis at YVR"
Great hotel. You would never know that you were at an airport. The gym is amazing, the food perfect and the staff is top notch.Genuine Expedia guest review
"Comfortable, clean stay"
Arrived late to Vancouver airport and made a last minute reservation. The staff was great expediting my check in so I could get to my room faster. The room was nice and clean. Although it was a short stay, it was a nice stay.Genuine Expedia guest review
"Fabulous hotel!"
I would return in a heartbeat! Stayed here over Halloween. The housekeepers were in costumes and were so sweet! There was a pumpkin carving contest and a costume contest for kids in the airport! It was so festive! I tried to tip the man who brought us our supper to our room but he declined. He told me it was already included in the delivery charge. […] I appreciated his honesty! I usually travel with my own pillow too but didn't want to take up that much space in my suitcase this time. I was fine with their pillows and beds. They are absolutely the most comfortable I have ever slept on! The tenderloin steak melts in your mouth and the vegetables are cooked perfectly- not wilted or mushy. We watched the planes land from our window but you can't hear them!!! The blackout curtains do the job too well! I had to leave the inner bathroom light on at night! Read moreGenuine Expedia guest review
"Nice but"
We had to stay here due to a flight disruption. For convenience it cannot be beat. For price . . . it is over priced. Had time permitted we would have looked for something far more reasonable.Genuine Hotels.com guest review
"Always convenient & comfortable for a quick visit"
The best advantage of this hotel is the commute from airport. Nothing can beat going 100m to the hotel when you land late in the evening after a delay with a connecting flight. The Fairmont-level service is followed with friendly staff and a fantastic fitness center and gym. My only pet peeve is there is not 'Fairmont' labeled bottled water in the room […] like in some Fairmonts (Bermuda, Dubai). This is not normally a problem but there isn't bottled water in the gym, either, so if you are running on the treadmill, you either need to buy the bottle of Evian in your room and try to fit it on the treadmill (it doesn't stay) or keep getting off the treadmill to poor a cup of water. Either the room or the gym should have complimentary bottled water. Read moreGenuine Hotels.com guest review
"The dusty hotel room!!!"
The room is dusty.(the 3 room we stay in) After the issue i have bring up and the room cleaner do the good cleaning one of the room. It is better. But I request the next to do the all 3 room the same procedure as last night they did. But they did not do it. The hotel need to find a solution for it. I have toll the manager what is the problem and I hope […] she will bring it up to the management and provide a better and quality service to all the gust visit in the future. Read moreGenuine Hotels.com guest review
We stayed in this hotel the night before an early morning flight because (a) Fairmont hotels are always nice and (b) the convenience of the hotel being actually inside the airport. Our room was on the 10th floor. The bed itself was very comfortable, although I wasn't crazy about the wimpy pillows. The room was clean and very spacious, with a large bathroom […] with separate bathtub and big shower. If you're worried about noise from the flights, don't be - we could barely hear them taking off even though you can see them outside your window. I don't think this airport has many (or any) flights that leave after 11pm, and even if they did, the noise is not enough to keep me awake and I am a very light sleeper. Staff was very friendly. The hotel has several bars and restaurants and a large room service menu with gluten-free and macrobiotic choices if you're willing to spend the money. If not, you can always go to one of the restaurants or the food court inside the airport. There is also a kiosk beside the front desk where you can check-in electronically for your flight on multiple airlines. We would definitely stay here again. Read moreGenuine Hotels.com guest review
"Very nice hotel"
Very nice hotel, but for this price you would think you would not have to pay for internet access.Genuine Expedia guest review
It's so nice, the view and the service.Genuine Hotels.com guest review
"Ideal location at the Vancouver airport"
It made out morning flight stress free to be at a hotel that was inside the airport. The room was clean and well appointed, but not luxurious. I did not like that there was no solid wall between the bathroom and bedroom area so that you could hear the other person in the bathroom very easily. Lighting was hard to find and adjust. The walls seemed thin […] and there was noise in the hallway. Service was very good and very courteous. We had a room service very late dinner as we arrived quite late. The soup was delicious. The sandwich was mediocre and very very expensive. Read moreGenuine Hotels.com guest review
"Nicest hotel ever!"
This is the nicest hotel ever hands down. If you need to stay at the Vancouver Airport, treat yourself to one of the nicest hotel stays you will ever have.Genuine Hotels.com guest review
"Worth every penny!"
I would stay here again in a heart beat. It was amazing to take elevator down to terminal. The room service was excellent and the beds were amazing.Genuine Hotels.com guest review
Response by The Fairmont Vancouver Airport Thank you for your review. We are happy to hear that you enjoyed the convenience of location, room service and comfortable beds. We have shared your kind comments with our teams. We look forward to welcoming you back soon.
"Hotel at the Vancouver International Airport"
Very convenient, luxurious hotel for those leaving Vancouver in the early hours.Genuine Expedia guest review
"End of vacation layover"
Decided to spend a day in Vancouver and take a morning flight instead of the red eye back to Cleveland. We expected to drop off our luggage after a 10am arrival and spend a day in the city, but when we arrived the room was already available, and the bed was extremely comfortable after a week of touring Alaska. Ended up spending most all of the day in […] the room relaxing. Read moreGenuine Expedia guest review
Enjoyed it very much. One of the best hotels I have stayed in.Genuine Hotels.com guest review
So convenient to walk from the lobby into the airport departures. Even had a Keurig machine in the room! Would definitely stay here again the next time I have a departure from Vancouver.Genuine Hotels.com guest review
"Very Convenient!"
this hotel is pricey, but very nice and VERY convenient. It is literally INSIDE the airport. For early morning flights out of YVR, nothing could be easier.Genuine Expedia guest review
"air flight ovrnight"
one of the perks to a two day passageeGenuine Hotels.com guest review
Fabulous shower. Very convenient. Walk in freezer for fishermen like myself. Service is always excellent. Gold floor standards totally worth it. Loved the canapés.Genuine Hotels.com guest review
"close to my airline"
wonderfulGenuine Hotels.com guest review
"Nice hotel, convenient to the airport"
Nice hotel, comfortable rooms. Although basically in the airport, completely soundproof- never heard any planes. The staff was all very friendly.Genuine Expedia guest review
"Exceptional Service"
Exceptional service, not just by airport hotel standards (where guests might expect to sacrifice service for convenience) but by any good hotel standard. Staff are genuinely interested in my satisfaction. We make this hotel a part of our 3-times-a-year vacations from Texas to our Vancouver Island condo.Genuine Hotels.com guest review
"Quiet hotel at the airport"
If you need to stay at the airport - this is the place. Terminals attached to the hotel. Plus it has a lovely bar/casual dining area as well as a very nice restaurant. We visited both and were not disappointed. The rooms are large and comfortable. Only issue is privacy in the bathroom - doors are louver and the sounds travel.Genuine Expedia guest review
"Once again they want you to pay for WIFI."
Once again we went to a Canadian Hotel that promoted WiFI only to find that we had to pay for WIFI Access, we passed on paying and went downstairs to the Vancouver Airport and got free WIFI access. You are locked into the Hotel if you do not have a car.Genuine Expedia guest review
"Nice, literally right at the airport."
Probably one of the best hotels I've ever stayed. Loved everything about it. So convenient how it's connected to the airport.Genuine Expedia guest review
"flying into canada"
staff helpful, room was very comfortable. location for us traveling from US and dealing with lots of luggage and a tired 5 year old was excellentGenuine Expedia guest review
"Great value for air travelers."
The world needs more hotels located directly at airport terminals, without the hassles of shuttles. Fairmont YVR is a terrific convenience for people who are flying in late or flying out early. The rapid transit line to downtown means you still have access to the city. The hotel was fancier than I needed, but the price was competitive for Vancouver. […] No complaints at all! Read moreGenuine Expedia guest review
"Excellent Hotel"
Stayed at this hotel for one night. My daughter and I had an early flight out of Vancouver and this was the ideal place to stay. Every member of staff involved was courteous and helpful. They could not do enough for you. We ate dinner at the hotel and it was also excellent. Would highly recommend this hotel to all travelers.Genuine Expedia guest review
"perfect hotel to use while travelling by air"
Doorman was very curtious and helpful. Reception clerk ,cheerful efficent and gave good information of hotel, including breakfast info and asked if we were in need of anything. Found our room comfortable , clean and quiet. Bellhop, was very helpful as well. I would reccomend this hotel to anyone travelling, wanting to get any time type of flight. We […] will certainly keep this hotel in mind if travelling again in the Vancouver area. Read moreGenuine Expedia guest review
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the fairmont always lives up to expectationGenuine Hotels.com guest review
"great location for business traveler"
clean and comfortable. Most importantly to me it was convenient to the airport.Genuine Expedia guest review
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This is a quiet, very comfortable hotel that allows you to relax, sleep soundly and still catch an early flight.Genuine Hotels.com guest review
"Unparalleled Service"
We make this a regular stop, 3 times a year before flying home after vacationing on Vancouver Island. The high level of service at this airport hotel continues to exceed our expectations each time we stay. Genuinely friendly and courteous staff go out of their way to make sure our stay is perfect. Rooms are very clean, comfortable and well-equipped […] . Read moreGenuine Hotels.com guest review
"When you need a hotel right now!!"
We arrived at midnight; desk clerk helped me make arrangements for next day's transportation by taxi to our next destination and had the information for me by the time we got to our room. In spite of being right beside the runways (great views) the hotel is very quiet. Wish we could have spent more time enjoying this convenient hotel.Genuine Expedia guest review
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Hotel staff make you feel wanted and are very friendly, it would be nice to have the same level of professionalism in hotels in the UK.Genuine Expedia guest review
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The invention provides a reagentless assay kit for analyte in a sample comprising a modular affinity assembly including at least one sensor unit comprising a ligand having binding affinity for the analyte (affinity module) operatively associated with a reporter probe (reporter module) responsive to changes in the sensor unit induced by analyte/receptor complex formation by transduction of a characteristic detectable signal. Assays employing the modular assembly are also provided.
1. A modular assembly for affinity separation of an analyte from a sample comprising:
an affinity support comprised of a metal; and
at least one sensor unit comprising:
an affinity module, said affinity module having a binding affinity for said analyte; and
a signal-competent reporter module, said signal competent reporter module being directly or indirectly signal-responsive to complex formation between said affinity module and said analyte, wherein said signal competent reporter molecule is immobilized on said affinity support, and said signal-competent reporter module being spaced from said affinity support such that said complex formation provides a change in the distance between said signal-competent reporter module and said affinity support.
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11. An assay kit for detection of an analyte comprising the modular assembly of one of claims 1-10.
12. A method for detecting an analyte in a sample comprising exposing the modular assembly of one of claims 1-10 to the sample and detecting a signal from the reporter module.
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This application makes reference to PCT Application No. PCT/US98/06871, entitled �Modular Assembly for Reagentless Affinity Separation and Detection of Analyte,� filed on Apr. 10, 1998, and published in the English language on Oct. 22, 1998, which claims priority to U.S. Provisional Patent Application No. 60/043,501, entitled �Modular Assembly for Reagentless Affinity Separation and Detection of Analyte,� filed Apr. 11, 1997. The entire disclosures and contents of the above applications are hereby incorporated by reference.
The inventions described herein relate to the affinity separation and detection of analytes in a sample. In particular, they relate to reagentless binding assays which permit contemporaneous (real time) monitoring of analyte concentration, detection of analyte at very low concentrations, and which are easily generalized for detection and quantitation of a host of different analytes.
1. Field of Art
The assays of the invention employ unique modular affinity assemblies as affinity matrices for affinity separation, detection, and quantitation of analyte. The assembly includes at least one sensor unit, each comprising at least one anti-analyte receptor (affinity module) and at least one signal-competent reporter probe (reporter module) signal-responsive to events associated with analyte/receptor complex formation. While the sensor units are exemplified herein as optical sensor units comprising luminescent reporter probes (luminophors), the use of binding-sensitive reporter probes which transduce non-optical signals such as electron transfer signals such as electron transfer and radioactivity in the modular assemblies of the invention are also contemplated.
The inventions are particularly useful for the detection of very dilute concentrations of analyte (in the ng/ml-pg/ml range and lower), and for continuously reporting changing concentrations of analyte in �real time,� (i.e., contemporaneously with the change in concentration), without further manipulation. Readings of the transduced signal can easily be taken at locations remote from the point of analysis, thus permitting, for example, constant monitoring of environmental air and water for pollutants, from afar.
Other applications for the assays of the invention include the clinical detection and real-time monitoring of trace biochemicals in body tissues and fluids, thus permitting, for example, the diagnosis and monitoring of biochemical markers of disease; the detection of trace amounts of hazardous (bio) chemicals in the environment such as those from medical, radioactive or industrial waste; the detection of pathogens (e.g., microorganisms and viruses or their toxins) in minute quantities in the atmosphere (including closed environments such as health-care facilities) or water supply; the detection of explosive materials; and the detection of proscribed substances such as controlled drugs.
2. Discussion of Related Art
Numerous assays based upon affinity chromatography are well-known for the detection of analyte in a sample. Typically, such assays involve immobilizing an analyte-specific, labelled ligand on a support to form an affinity matrix; contacting the immobilized ligand with a fluid sample potentially containing analyte; and detecting and/or quantitating bound analyte. Competitive binding assays, dependent upon competition between analyte and a labelled analyte analog for ligand binding sites are more complicated, additionally requiring labelled analyte analogs for initial saturation of matrix-bound ligands. Sandwich-type binding assays also additionally require labelled secondary ligands for sandwiching analyte bound to the primary immobilized ligand.
These classic assays have several drawbacks which make them unsuitable for achieving the objectives of the present invention. In both assays, the equilibrium eventually established between the analyte and immobilized ligand reflects a static sample analyte concentration, but fluctuations in the concentration cannot be detected without further addition of reagents, viz., labelled analog or labelled secondary ligand. Further, the assay is not reversible without time-consuming regeneration of the matrix, and even so this is not always possible. The present invention obviates the need for secondary labelling steps that require time for mixing, reaction, and flushing of excess reagent prior to detection and quantitation of analyte, and permits real-time continuous monitoring of analyte concentration without matrix regeneration.
The development of reagentless fluorescence-based sensors has long been a goal in this art. Several reagentless techniques that allow nearly real-time monitoring of chemicals have been developed, but all have been limited to a narrow range of analytes and receptors. In many of these techniques, the receptor itself is a fluorescent molecule (e.g., a metal ion chelator) whose fluorescent properties (e.g., emission intensity, emission wavelength, or lifetime) change upon analyte binding. Accordingly, a large effort has been expended on developing fluors that also have analyte recognition properties.
For example, a variety of cation-specific fluorescent dyes are commercially available that exhibit an intensity enhancement or a shift in their fluorescence emission spectra upon ion binding,. In addition to pH sensitive dyes, there are several known fluorescent probes that specifically chelate ions such as Mg+2 (e.g., furaptra), Zn+2 (TSQ), Na+ (SBFI), K+ (PBFI), and Ca+2 (EGTA-AM) for use in the detection of relevant chemicals (Haugland, R. P., Handbook of Fluorescent Probes and Research Chemicals, Molecular Probes, 1996). These dyes have found wide application both in environmental sensing applications and in biochemical studies. The methods employed for detection of analyte with these dyes, however, have the disadvantage of requiring a specific receptor for the analyte that is also fluorescent, and whose fluorescent properties change upon analyte binding. Thus, they cannot be generalized to arbitrary analytes. Similar restrictions also apply to other optical sensor units used to detect excited state fluorophors or phosphors (Meier, B.; Werner, T.; Klimant, I.; Wolfbeis, O. S., �Novel Oxygen Sensor Material Based on a Ruthenium Bipyridyl Complex Encapsulated in Zeolite-Y: Dramatic Differences in the Efficiency of Luminescence Quenching by oxygen on Going, From Surface-Adsorbed to Zeolite-Encapsulated Fluorophores,� Sensors & actuators B 2: 240, 1995).
A related reagentless approach is the use of environmentally-sensitive dyes with proteins, polymers, and in molecular assemblies (Lundgren, J. S.; Bright, F. V., �Biosensor for the Nonspecific Determination of Ionic Surfactants,� Anal. Chem. 68: 3377, 1996). Dyes such as 6-propionyl-2-dimethylamino-naphthalene (prodan) and 6-dodecanoyl-2-dimethylamino-naphthalene (laurodan) have been used for binding of a wide range of analytes. These dyes incorporate both electron donor and electron acceptor moieties that result in a large dipole moment in the excited state (Haugland, R. P., Handbook of Fluorescent Probes and Research Chemicals, Molecular Probes, 1996). Consequently, the emission spectra of these dyes are extremely sensitive to the polarity of their environment. For example, the emission maximum for prodan varies from about 380 nm when the dye is in a nonpolar environment (e.g., cyclohexane), to about 520 nm when the dye is in a polar environment (e.g., water) (Haugland, R. P., Handbook of Fluorescent Probes and Research Chemicals, Molecular Probes, 1996). In typical sensing protocols, these dyes are incorporated into polymers that swell in the presence of certain types of low molecular weight analytes (Barnard, S. M.; Walt, D. R. �A Fibre-Optic Chemical Sensor with Discrete Sensing Sites,� Nature 353: 338-340, 1991). The degree of swelling of the polymer is directly proportional to the amount of analyte that has been imbibed by the polymer, and the resulting change in the polarity of the environment of the dye affects its fluorescence emission properties, which can then be correlated to analyte concentration. Similar techniques have been used to study the dynamics of cell membranes. For example, the effects of drugs, anesthetics, extracellular proteins, and metal ions on membrane dynamics and structure have been monitored by the incorporation of fluorescent probes such as diphenylhexatriene into the membrane structure (Haugland, R. P., Handbook of Fluorescent Probes and Research Chemicals, Molecular Probes, 1996). Although these general methods for fluorescence-based transduction allow reagentless monitoring of a variety of types of analytes, it is not broadly generalizable and it also lacks analyte specificity, as many background substances can cause changes in the environment of the fluorescent molecule that lead to confounding spectral data.
One type of reagentless, fluorescent sensor unit having a high degree of specificity comprises a specific binding receptor protein modified by covalent incorporation of reporter fluors. These sensor units are engineered so that emission properties of the fluor are changed with analyte/receptor binding. As an example, three general transduction mechanisms in which binding of analyte to the modified receptor results in a perturbation of the fluorescence emission have been identified (see Case et al and Sohanpal, K.; Watsuji, T.; Zhou, L. Q.; Cass, A. E. G., �Reagentless Fluorescence Sensors Based Upon Specific Binding Proteins,� Sensors & Actuators B 11: 547, 1993). These are (1) perturbation of the fluor either through direct interaction with the analyte upon binding, or through indirect interaction via the receptor upon binding; (2) perturbation of the fluor caused by conformational changes in the receptor protein upon analyte binding that results in a change in the local chemical environment of the fluor; and (3) perturbation of the fluor caused by a change in the aggregation state of the receptor protein upon analyte binding. Although these transduction mechanisms are reagentless, reversible, and specific, they are too specific to be generalizable, as the fluor must be preattached to each protein receptor at a site that typically differs depending upon the protein receptor selected.
FIG. 1: Dependence of the relative fluorescence lifetime on distance of separation between a fluorophore and a metal surface, as estimated by Kuhn's approximation theory. (Figure is adapted from Chen, H. C.; Frank, C. W., �Fluorescence Probe Studies of Self-Assembled Monolayer Films,� Lancimuir 7: 1791, 1991).
FIG. 2: Attenuation of fluorescence emission intensity of a rhodamine containing modular assembly formed upon analyte (Cs+) binding.
In this Example, the molecular assembly is a self-assembled monolayer (SAM) formed by the reaction of HS(CH2)10C(O)NH(CH2)2O)2(CH2)2NHC(S)NH�Rh wherein Rh=rhodamine, with the surface of a thin gold film. The cesium analyte binds to the oligo(ethylene glycol) moiety NH(CH2CH2O)2CH2CH2NH in the assembly, resulting in a change in the structure of the molecular assembly which reconforms the rhodamine to a less luminescent conformation.
FIG. 3: Transduction in Modular Assemblies.
In FIG. 3A, R represents a receptor module and L1 and L2 reporter modules. In one embodiment of the present invention, L1 and L2 may be excimer couples where binding of the analyte results in excimer emission. In another embodiment of the present invention L1 may be a luminophore and L2 is an effective quencher (through electron transfer or resonance energy transfer systems) and binding of the analyte results in decreased emission. Yet another embodiment of the present invention may include a reporter where L1 and L2 are capable of photoinduced charge transfer and binding of the analyte can result in either increased or decreased emission depending on the identity of L1 and L2.
FIG. 3B: Binding of the analyte results in displacement of an effector module (R) away from the luminophore, increasing emission. R is, for example a quencher or is linked to a quencher.
FIG. 4A illustrates in schematic form the reagentless transduction for specific binding of a model analyte (streptavidin) to a modular assembly on a metallic substrate (gold) in wbich biotin is the model receptor for stretavidin.
FIG. 4B is a graph of fluorescent spectrums of rhodamine in which open circles labeled �control� represent a fluorescence spectrum of reporter rhodamine from the assembly that has not been exposed to streptavidin, open diamonds labeled �streptavidin� represent a fluorescence spectrum of reporter rhodamine after exposure to streptavidin, and filled triangles labeled �BSA� represent a fluorescence spectrum of rhodamine after exposure to a receptor that does not bind to biotin
FIG. 5A illustrates in schematic form a reagentless transduction of reversible binding of a model analyte (antibiotin to a modular assembly on a metallic substrate (gold) in which biotin is the model receptor for antibiotin.
FIG. 5B is a graph of fluorescent spectrums for experiments based on the reagentless transduction of FIG. 5A in which open circles labeled �control� represent a fluorescence spectrum of reporter rhodamine from an assembly that has not been exposed to antibiotin, open diamonds labeled �antibiotin� represent a fluorescence spectrum of reporter rhodamine after exposure of antibiotin, and filled triangle labeled �biotin� represent a fluorescence spectrum of reporter rhodamine after elution of antibiotin from the assembly surface by exposure to excess biotin.
The invention provides a reagentless binding assay kit for analyte (ligate) in a sample comprising a sensor unit including a receptor or other ligand having binding affinity for the analyte (affinity module) operatively associated with a reporter probe (reporter module) responsive to changes in the sensor unit induced by analyte/receptor complex formation by transduction of a characteristic detectable signal.
In one embodiment, the invention provides a solid state, modular affinity matrix assembly including at least one such sensor unit immobilized on a solid affinity support. The modules of the assembly are physically constrained (ordered) by the assembly so that formation of bound analyte/receptor complexes disturbs the orderedness of the assembly and to cause physical, electrical and/or clinical changes directly or indirectly reportable by the reporter module. Luminescent reporters (luminophors) sensitive to changes in the orderedness of the modular assembly caused by the formation of receptor/antireceptor complexes are especially useful in assays requiring high sensitivity for bound analyte, as these reporters used in accordance with the invention are capable of detectably signalling the presence of as little as a single bound analyte.
The invention further provides assays employing the modular assembly of the invention.
The modular assemblies of the present invention are affinity matrices including de minimis one sensor unit comprising a receptor (affinity module) and a reporter probe (reporter module) immobilized on an affinity support. The affinity module comprises any affinity ligand known in the art capable of forming a bound complex with a target analyte. For many applications, it will be desirable for the receptor to have a high specificity and affinity for the target analyte; however, receptors having broader affinities, such as those cross-reactive with related bacterial species, may be desirable for other applications. Contemplated analytes are myriad, and require only a complementary ligand to be a suitable target for assay according to the invention. Particular analytes include macromolecules such as proteins, whole cells, immunogenic peptides, and heavy metals.
The reporter module comprises any known signal-competent moiety capable of transducing a detectable characteristic signal in response to stereochemical events accompanying formation of bound receptor/analyte complex. These events include reporter perturbation upon binding of analyte caused by direct or indirect interaction with the analyte; conformational changes in the receptor; changes in the aggregation state of the receptor; changes in the chemical or dielectrical environment of the reporter module; and displacement of the reporter module relative to an effector module incorporated in the assembly. As noted above, particularly preferred reporter modules comprise optical reporters including luminophors and chromophors. These modules have emission properties modulatable in response to analyte/receptor complex formation, such as intensity, color, and lifetime of radiation; wavelength distance and frequency, shifts in spectra, and changes in polarization. Complex formation is readily detected and quantitated by a change in such emission properties by known techniques. Other reporters comprise enzyme- or redox-based electron transfer systems known in the art for their ability to transport detectable signals.
The reporter modules typically respond to complex formation by a fluctuation in signal; however, the modules may also have an on/off switch, responding by turning on a previously undetectable signal, or by turning off a detectable signal.
Effector modules are optionally incorporated into the modular assemblies for mediation of the signal response of the reporter module to affinity complex formation. Exemplary effector modules comprise, for example, excimer couples, collisional quenchers and resonance energy transfer donors or acceptors which are directly affected by receptor/analyte complex formation and in turn effect corresponding signal changes in luminophor or other reporters. Other effector modules include components such as metallic particles or films having dielectric properties which change the luminescent properties of luminophor reporters upon displacement of the luminophors relative to the dielectric material upon complex formation.
Thus, the detectable signal response may be 1) signal initiation and transduction (preparation), 2) signal extinction, or 3) signal fluctuation.
The module components of the assembly are disposed on the affinity support in operative relationship so that physical, electrical, or chemical events accompanying complex formation are transmitted to the reporter module, either directly or via an effector module, and result in a detectable signal of complex formation. The affinity module, while associated with the other modules of the invention within the assembly, is either affixed to the support so it is spaced from the reporter module, or the reporter and affinity modules are linked. Each of the sensor units of the assembly comprises at least one receptor and one reporter module immobilized on the support; the modules may be the same or different. The receptor module may differ from unit to unit, but be associated with the same reporter module, and vice versa; each unit may have the same reporters and receptors; or both reporter modules and receptor modules may differ from unit to unit. For example, a first sensor unit may include an antibody to virus A as receptor with an associated fluor reporter, while a second sensor unit may include an antibody to virus B as receptor with an associated phosphor reporter. Such a modular assembly is particularly suitable for sampling an environment for a variety of possible analytes by inducing different signals for different bound analytes. The received signals may then be read and quantitated for specific analyte. Possible subunit constructs include:
The support comprises a solid, optionally rigid, substrate selected from materials known in the art for affinity ligand immobilization. Criteria for such supports are known; for example, the support should be free from extraneous ion-exchange sites, should not promote non-specific binding, should be mechanically strong enough for the contemplated application, should be biologically and chemically inert, and be adapted for ligand immobilization. The support should also be stable under reaction conditions (e.g. for ligand immobilization) and in its intended environment for use. Useful supports for module immobilization according to the present invention comprise organic polymers, metals, semiconductors, and ceramics. The support may comprise a mixture of these materials, such as an organic polymer layer bonded to an inorganic layer to provide attachment sites for certain affinity modules, or a non-conducting oxide layer on a metallic or other substrate. For most applications, the affinity support will comprise a relatively thin film or composite film as further described infra.
Modular engineering of reagentless analyte sensor units according to the invention includes the manufacture of a minimum combination of two modules having the separate functions of (a) analyte binding affinity (affinity module) and (b) recognition-responsive signalling (reporter module). The sensor units are engineered into the assembly so that the binding event between the receptor and analyte affects the signalling properties of the reporter module sufficiently to permit detection of the event. The modular design concept is attractive because it is useful with a variety of receptors and their target analytes. Thus far, however, it has been only used to generate indicators for binding of heavy metal ions in solution.
Modular Assemblies. The modular assemblies of the invention are prepared by covalent or non-covalent attachment of component modules, to a solid affinity support or substrate such as a metal, semiconductor, ceramic or synthetic or natural organic polymer. They are solid-state assemblies. herein distinguished from other affinity matrices having affinity supports comprising fluid, free-standing membranes such as lipid bilayers, vesicles and Langmuir films. The solid state assemblies of the invention are engineered for sufficient structural stability so that direct or indirect perturbation of the reporter module to cause signal changes reliably reflects complex formation rather than extraneous events. The supports comprise known materials used in the art for the immobilization of affinity ligands. These materials include self-assembled monolayers (SAMs); polymeric brushes and grafted polymers. Materials that are ordered assemblies (e.g., SAMs of alkylthiols on noble metals and some semiconductors, of silanes on oxides, or of carboxylic acids on basic oxides, polymer brushes and certain grafted polymers, are herein distinguished from those that are normally relatively disordered such as polymer gels and surface-adsorbed insoluble polymers. There have been only a few reports on the use of such assemblies for reagentless chemical sensing (Motesharei, K.; Myles, D. C., �Molecular Recognition in Membrane Mimics: A Fluorescence Probe,� J. Am. Chem. Soc. 116: 7413, 1994), and no reports of transduction mechanisms based on changes in the orderedness of modular assemblies upon analyte binding and detecting.
Luminescence-Based Sensor units. A number of virtues make measurement of changes in luminescence (e.g., fluorescence, phosphorescence) properties a desirable method of transduction in affinity-based sensor units. Under appropriate conditions, luminescence spectroscopy can be used to probe extremely low concentrations of analyte; changes in fluorescence spectra and lifetimes can even be detected from a single molecule (Ambrose, W. P.; Goodwin, P. M.; Martin, J. C.; Keller, R. A. �Alterations of Single Molecule Fluorescence Lifetimes in Near-Field Optical microscopy,� Science 265: 364, 1994). Luminescence can essentially function as an amplification mechanism, as a single luminophor can be subjected to multiple cycles of excitation and emission, each of which reflects the binding state of a receptor with which the luminophor is associated. Other useful properties of luminescence include the fact that intensity of absorbance and emission at various wavelengths, luminescence polarization, and excited state lifetime, are all properties of the luminophor that are, in general, sensitive to the chemical and electrical environment of the luminophor. Hence, these properties can all be used to advantage in a multi-measurement analysis of the binding state of a receptor with which the luminophor is associated for real time monitoring of analyte. Recent advances in measurement of excited state lifetimes, for example, are particularly relevant to affinity complex sensing, as they have led to the development of compact instrumentation for measuring phosphorescence and fluorescence lifetimes in schemes that are relatively insensitive to external interferences such as spurious light sources and photobleaching effects (Meier, B.; Werner, T.; Klimant, I.; Wolfbeis, O. S., �Novel Oxygen Sensor Material Based on a Ruthenium Bipyridyl Complex Encapsulated in Zeolite-Y: Dramatic Differences in the Efficiency of Luminescence Quenching by Oxygen on Going, From Surface-Adsorbed to Zeolite-Encapsulated Fluorophores,� Sensors & actuators B 29: 240, 1995).
As described above, the present inventions are based on modular assemblies that incorporate affinity and reporter modules selected and disposed on the support so that binding of an analyte to the receptor (affinity) module results in one or more changes in the architecture (order) of the molecular assembly to which the luminescent or other properties of the reporter module are sensitive. Examples of such changes are: (1) a change in the structure of the modular assembly resulting in a change in the average distance between the luminescent centers of a luminophor module and a solid substrate having appropriate dielectric properties (e.g., n and k) so that the original luminescent properties of the luminophor are changed upon analyte binding; (1) (b) a change in the structure of the assembly resulting in a change in the average distance between an electroactive reporter probe, e.g., a redox reporter such as viologen or a redox enzyme such as horseradish peroxide; (2) a change in the structure of the modular assembly so that the transport properties of quenchers such as oxygen are modified upon analyte binding; (3) a change of the structure of modular assemblies so that the luminescent properties of a luminophor reporter are indirectly affected by effector modules built into the modular assembly such as excimer couples, collisional quenchers and resonance energy transfer donors or acceptors; (4) a change of the structure of modular assemblies so that the luminescent properties of luminophor reporter modules such as fluors and phosphors are directly affected by the analyte, for example, through collisional quenching, photoinduced electron transfer, or resonance energy transfer. Several examples of covalently-linked receptor and transducer modules based on photoinduced electron transfer (PET) are known; (see, e.g., Prasanna de Silva, A.; Gunnlaugsson, T.; Rice, T. E., �Recent Evolution of Luminescent Photoinduced Electron Transfer Sensors,� Analyst 121: 1759, 1996; Fabbrizzi, L.; Licchelli, M.; Pallavicini, P.; Sacchi, D.; Taglietti, A. �Sensing of Transition Metals Through Fluorescence Quenching or enhancement: A Review,� Analyst 121: 1763, 1996, incorporated herein by reference). Such mechanisms permit the generalization of the present affinity matrices to permit the facile detection of a host of analytes.
A particularly significant aspect of the invention is the enablement of binary or higher order modularity of function provided by the inventive modular concept. In addition to the affinity (receptor) and reporter (e.g., luminophor) modules in the assemblies, modules performing additional functions may also be included. For example, for sensing in biological fluids an anti-fouling module comprising, for example, polyethylene glycol or a highly-hydrated polysaccharide can be included to eliminate biofouling of the affinity matrix.
Within a distance approximately equivalent to the thickness of the modular assemblies envisioned here (typically about �10 Å-1 μm), the radiative emission of fluorophors and phosphors is known to be very sensitive to their separation from metals and other materials (Pockrand, I.; Brillante, A.; Mobius, D., �Nonradiative Decay of Excited Molecules Near a Metal Surface,� Chemical Physics Letters 69: 499-504, 1980). For example, FIG. 1 illustrates the dependence of fluorescence lifetime on distance of separation from a metallic surface, as estimated by Kuhn's approximation theory. For rhodamine (λemission �600 nm), and n1=1.45 (the approximate value for close-packed hydrocarbon assemblies), a value of 1.0 on the X-axis corresponds to a separation distance of �65 nm. This separation is in the midrange of thicknesses for the thin film molecular assemblies envisioned here.
To a first approximation, the components of the complex index of refraction (n and k) of the substrate can be used, together with the separation distance and the orientation of the fluor or phosphor relative to the substrate surface and the roughness of the substrate surface, to estimate how strongly the surface will influence the lifetime and emission intensity of the reporter (Chen, H. C.; Frank, C. W., �Fluorescence Probe Studies of Self-Assembled Monolayer Films,� Langmuir 7: 1791, 1991). Several new types of synthetic molecular assemblies are described below that can take advantage of these phenomena in the transduction of binding of an analyte to a modular assembly. In the following Examples a synthetic (bio)organic assembly is formed either directly on a metallic surface (e.g., gold, silver), on an nonconducting oxide on a metallic surface or on any other solid support (e.g., semiconductor, oxide, organic polymer) that can function either as a spacer from the metallic surface or, if designed with the appropriate dielectric properties as known in the art, also as the module that perturbs the signalling properties of the reporter module. The following Examples demonstrate transduction by direct measurement of fluorescence lifetime, or by measurement of fluorescence intensity and/or polarization that may correlate with fluorescence lifetime. Very thin films of metals or other materials that are transparent to excitation and emitted light can be formed on supports for compact sensing platforms based, for example, on evanescent wave excitation.
In addition to luminescent reporters, other reporters sensitive to changes in their distance from a metallic or other support may be employed. These include redox reporters such as electroactive chemical and biochemical compounds, e.g., ferrocene, viologen, and enzymes that catalyze redox reactions.
Various receptors, including some based on native biomolecules (Sohanpal, K.; Watsuji, T.; Zhou, L. Q.; Cass, A. E. G., �Reagentless Fluorescence Sensors Based Upon Specific Binding Proteins,� Sensors & Actuators B 11: 547, 1993) and some based on synthetic molecules (Fabbrizzi, L.; Licchelli, M.; Pallavicini, P.; Sacchi, D.; Taglietti, A. �Sensing of Transition Metals Through Fluorescence Quenching or enhancement: A Review,� Analyst 121: 1763, 1996), exhibit changes in the conformation of their molecular chains upon binding of an analyte. On co-localization of these receptors with luminophors, electrophors (electron-transfer sensitive probes), or other reporters, in a modular assembly in the vicinity of a metal or other film that influences the emission properties of the reporter, the resulting change in receptor conformation results in a change; e.g., in the average distance between a luminophor reporter and the metal or other surface, and thus in a change in detectable luminescent properties of a luminophor. Any receptor change resulting from complex formation that influences the relative distance between the luminophor and the surface or otherwise affects its dielectric properties is utilizable for transduction according to the invention. FIG. 2 illustrates this transduction mechanism. In this illustration, a fluor covalently linked to the receptor is used to transduce a signal responsive to analyte binding.
Several other transduction mechanisms based on changes of assembly architecture are contemplated:
decrease in average separation of luminophors from a metallic or other surface (e.g., as a result of crosslinking or decrosslinking of the assembly upon analyte binding). Crosslinking and decrosslinking reactions can be induced by the presence of analyte through competitive dissociation. In this scheme, in the absence of analyte, the receptor is non-covalently bound to an analyte analog co-immobilized (e.g., covalently) in the molecular assembly. Upon exposure to analyte in solution, the receptor-analyte-analog couple is competitively dissociated owing to the formation of receptor-analyte complexes. In this embodiment, the receptor is labelled with a transducing probe [e.g., luminophor or redox probe (electrophor)], whose transduction properties are strongly influenced by the metallic or other support. Upon competitive dissociation via exposure to analyte (i.e., decrosslinking) the distance between the labeled receptor and the substrate changes (e.g., via diffusion). The sensor is made reversible by (1) linking the labeled receptor (e.g., covalently) through a flexible tether molecule (e.g., polyethylene glycol) to the support; or (2) by confining the receptor/analyte complex to the vicinity of the sensor surface by means of a selectively permeable membrane that does not allow transport of the larger receptor molecule (e.g., an antibody) but allows transport by the smaller analyte molecule.
increase in average separation of luminophors from a metallic or other surface (e.g., as a result of swelling of the modular assembly upon analyte binding). Swelling is usually caused by the excluded volume of the analyte, or by the removal of crosslinks by the analyte.
changes in average orientation of luminophors with respect to a metallic or other surface upon analyte binding.
Reagentless transduction can also be based on the use of quenchers capable of quenching excited states of a luminophor in the sample or sensing medium (e.g., O2 in air or dissolved in water). In these assemblies, binding of the analyte to a receptor results in a change in the modular assembly which inhibits the transport of a molecular quencher (e.g., O2) to the luminophor. Transport (e.g., diffusion) inhibition can be affected by, inter alia:
an occlusion of the surface of the assembly by the analyte (e.g., a protein);
a decrease in the porosity of the assembly,
a change in the solid state diffusivity in the modular assembly.
Several reagentless transduction mechanisms based on the concept of modular design of responsive molecular assemblies are shown in FIG. 3.
The modular incorporation of receptor and luminophor modules are disposed in close proximity in the assembly to cause a change in the photoemission characteristics of a luminescent reporter upon binding of the analyte to the receptor. Typical luminophor properties affected by the analyte include photoinduced charge transfer, resonance energy transfer, and changes in the local dielectric environment of the luminescent probe.
Examples are given in FIGS. 4 and 5 which schematically illustrate other specific and reversible reagentless transductions in the modular assemblies of the invention.
Patent Citations
Non-Patent Citations
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text:
That's why I make my batches .25 gal larger then my planned finish size. All the trub goes into the fermenter.
Yeah that's my current solution, increasing the batch size. Just curious if there are other things people are doing.
_________________________Fermenting: Golden Sour w/ECY01, Dark Sour w/2nd gen ECY01, plum wineBottle Conditioning: Leftover Grain Pumpkin Brown AleDrinking: Day Drinker Mild, Saison w/Brett, Berliner Weisse, Imperial Stout (4 variants)
Except standard trub management (irish moss, whirlpooling), sometimes I'm using cheese cloth. Sanitized by boiling and soaking in starsan. It fits perfectly into the big funnel. And wort is just poured through it. Always some tiny particles will find their way to the fermenter, but mostly what is left, it's a sludge from hops & cold break material
Anyone here tried going straight from a BIAB hot into a Sanke for a BIAB - No-Chill combo for max simplicity?
Did my first BIAB this weekend and i was shooting for 1.050 and hit 1.039. I used the recipe from here
My water temperature didn't change much after adding grains so I started mashing at 158. I didn't want to add cold water since I was pretty sure my amount estimate was going to be very close or high. Temp dropped about 1.5 degrees every 10 min.
I think part of the problem was I can't remember if I waited 10 minutes after getting to heating to 170, son was in the garage with me and it just slipped my mind to wait. So, basically after mashing, I raised temp to 170, then I think cut the heat, got pulley setup, then pulled it out, allowing it to drip for 15 min. Would it be normal to pick up that many gravity points from the 170 degree "sparge?" I also ended up leaving between .5 and a whole gallon in the kettle, most of which was trub, but some may have been viable wort. Ran out of room in the fermenter, hind sight i should've put it into 2 fermenters with a little more than 3 gallons in each.
Kinda sounds like you didn't get the evaporation you planned for if you ran out of room, which would definitely contribute to lower gravity.
After looking back at my plans, the recipe, and some calculations I think you're right. Pre-boil my gravity was 1.015, I had planned to do a 90 minute boil since I used Pilsner malt, but only ended up doing a 60 minute due to being distracted and just operating by habit from my first 6 or 7 brews.
The extra 30 min of boil would've "burned off" .5 gallon going by what I think my evaporation rate was. With a pre-boil of 1.015 and an OG of 1.039, looks like I was getting .012 per half hour. Add .012 to 1.039 and you get 1.051. So, maybe my estimation were pretty much spot on, just forgot to boil for an extra 30 minutes.


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Greg Warren brings himself and a cast of characters to the Louisville stage.
Former Louisvillian comes home for the weekend.
Tim Northern is rocking a full weekend of comedy at Laughing Derby
Local comedian Tom Mabe returns to Louisville for a good cause.
Comedy legend Joan Rivers returns to Louisville tonight.
Local comedy club supports not only their hometown team, but their community, as well.
there's something for everyone in Louisville this weekend.
where would I want to go?
Get ready as Robocop takes on his most formidable opponent...his friends.
Some thoughts on the passing of comedy legend Tim Wilson.
Moon Taxi was cleared for landing to rock a sold-out crowd on Friday night at Headliners Music Hall.
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Light Industrial, Clerical & Event Staffing
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We have a long history of filling positions with some of the top brands and companies in the Minneapolis and St. Paul metro area for over 30 years. We specialize in keeping our clients fully staffed and finding jobs for good people. We pre-screen, interview and train all of our employment candidates so they are ready for any position on the first day of work.
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text: Through offices in Wausau and Eau Claire, nearly 100 employees at Ruder Ware work together to provide exceptional service to our clients. Our attorneys provide counsel to many of the same clients that have been with the firm since its humble beginnings. An equal opportunity employer, Ruder Ware offers employees a professional environment in which they can grow and be challenged. At the largest law firm headquartered north of Madison, employees have an opportunity to work with a diverse client base, from privately held businesses to international corporations to individuals.


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PBs since age 60: 5k- 24:36, 10k - 47:17. Half Marathon- 1:42:41.
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MEC Trail Race Series - race one January 11 2015 Point Pleasant Park DNS
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Halifax Bluenose HM May 2015
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My next one is March 21 in Moab, Utah...I'm getting psyched!


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Maintenance Man II
After years of being out of the gigolo lifestyle, Malcolm has good looks, his rolodex, and his reputation. This time he vows that nothing will stand in his way of getting back on top and creating a franchise of escort services worldwide. He disguises his gigolo lifestyle as a nightclub owner in Miami -- the perfect cover to mix business with pleasure. But with high rewards come high risks.
Publisher: [S.l.] : Baisden Publishing, LLC, c2012.
Characteristics: 320 p. ;,24 cm.
Alternate Title: Maintenance Man two
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%% Image selected per Image Pickin' thread: http://tvtropes.org/pmwiki/posts.php?discussion=1334162598081780800
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->''"Grantville, West Virginia was the mold that produced Jeff Higgins. All things said and done, it was as good a mold as any and a better one than most."''
-->-- ''Literature/SixteenThirtyTwo''
This species has seldom been seen and naturalists have considered putting it on the endangered species list. The All-American boy is [[CaptainObvious male]], probably white, blond-haired and blue-eyed. He is marked by his love of baseball, by having a skill with rural machinery and hunting firearms beyond his years, and his propensity to emit sounds like "gee whiz". The All-American boy usually dwells in a QuirkyTown in the heart of {{Eagleland}} within which he is as [[FreeRangeChildren free as the air, pedaling everywhere on his bicycle]] or spending hours having innocent fun in his treehouse. He is naive but charming and always polite, and he treats his parents (who most likely are a StandardFiftiesFather and a HouseWife) and other elders with respect. He is probably a [[ScoutOut Boy Scout]] (or a Cub Scout if still in elementary school). If he has a sibling, it will be an [[BigBrotherWorship older brother to idolize]] or a little sister [[BigBrotherInstinct to protect]] -- perhaps both.
There are variants of this trope. The geeky variant is similarly characterized by ingenuity, self-reliance, and wholesomeness, but he applies his interest to at-home science experiments and the like. The high-school variety wears a letterman's sweater, plays football or baseball, and spends his off hours using his mechanical skills to restore an old car. You might have a "wilder" boy with a mischievous streak in him, but he'll grow out of it eventually and become the fine, upstanding man his parents raised him to be.
An All-American boy often gets a job as a KidDetective. If he joins the military when he grows up he will almost inevitably become a SouthernFriedPrivate. He may want to see more of the world and venture into TheCity, but expect him to return to his hometown, disillusioned with the citygoers' air of cynicism and greed. Your All American Boy will almost certainly marry his [[VictoriousChildhoodFriend high school sweetheart]], and then settle down to raise a family, with at least one son just like him.
The closest DistaffCounterpart would probably be GirlNextDoor.
[[folder: Advertising ]]
* Used in some commercials for Smuckers jams and jellies. Typically feature young boys (apparently the guys who would later found the company) on bicycles riding through orchards and playing together during TheFifties (or thereabouts).
* Ditto for the Blue Bell Ice Cream commercials, especially the radio variety.
[[folder: Art ]]
* NormanRockwell depicted many variations of him in his paintings, notably "A Day in the Life of a Boy," illustrations for the Boy Scouts of America, the Willie Gillis series, his illustrations for ''TomSawyer'' and ''HuckleberryFinn''... Really, it's hard to think of a Rockwell painting that doesn't involve him in some way.
[[folder: Comics ]]
* Steve Rogers (aka CaptainAmerica) pretty much fits the general personality of this trope, although he was a terrible athlete as a child, in large part due to his GeekPhysique. After he got the Super-Soldier Serum, he was able to embody the trope even more.
* Archie Andrews of ''ArchieComics'', especially in TheFifties and TheSixties.
[[folder: Fanfiction ]]
* If [[NationsAsPeople America]] shows up in one of the ''many'' ''AxisPowersHetalia'' HighSchoolAU fanfics, there's a pretty good chance he'll be this, for [[ExactlyWhatItSaysOnTheTin obvious reasons]].
[[folder: Film ]]
* Audie Murphy in the beginning of ''To Hell And Back''.
* The title character in ''MrSmithGoesToWashington'' is a grown-up example, as well as all the boys in his [[ScoutOut "Boy Rangers"]] group.
* ''Film/TheLeagueOfExtraordinaryGentlemen'' smuggled the probable TropeCodifier into its MassiveMultiplayerCrossover, despite this making no contextual sense.
* ''Film/BigFish'' is a great example. Quirky small town, baseball, etc.
* The Film/AndyHardy film series.
[[folder: Literature ]]
* TropeCodifier TomSawyer and HuckleberryFinn.
* TheHardyBoys.
* A pair of {{Kid Detective}}s in ''The Crow and the Castle'' by Keith Robinson.
* Jeff and the "four horsemen" in ''Literature/SixteenThirtyTwo'', who lean more toward the geeky variant in this case.
* Some of Creator/RobertAHeinlein's works are this RecycledInSpace.
* Henry Huggins in the ''RamonaAndBeezus'' books by BeverlyCleary.
* Galen Waylock in ''Literature/WarOfTheDreaming'' is an example, with the slight variation that he is a warlock trained to follow in the family AncientTradition of guarding humanity.
* Billy Coleman in ''WhereTheRedFernGrows''.
[[folder: Live-Action TV ]]
* Theodore "Beaver" Cleaver, from ''LeaveItToBeaver''.
* Opie Taylor and his pals on ''TheAndyGriffithShow''.
* Cory Matthews of ''BoyMeetsWorld'' is a modern example. He begins by caring about more baseball than anything and seeing his father as Superman. Since the show follows him from grade school to college, it gradually shifts from playing the trope straight to deconstructing it at times.
[[folder: Newspaper Comics ]]
* ComicStrip/DennisTheMenaceUS is closer to this than the name suggests, unlike his [[ComicStrip/DennisTheMenaceUK British counterpart]].
[[folder: Radio ]]
* Jack Armstrong, the All-American Boy.
[[folder: Theater ]]
* Biff in ''DeathOfASalesman'' is this as a kid. As he grows up, not so much.
* ''Theatre/ByeByeBirdie'': The number "A Healthy, Normal American Boy" describes Conrad Birdie as this in a series of BlatantLies.
[[folder: Video Games ]]
* Ninten from ''VideoGame/{{MOTHER|1}}'' and Ness from ''VideoGame/EarthBound'', both of whom are bat-wielding Everyman boys from a small town.
* Mike Jones, teenaged ace pitcher from ''VideoGame/{{Startropics}}''. His All-American-ness--contrasted with and found strange by the natives of the islands he's visiting--is a large part of the game's humor and tone.
* ''VideoGame/TeamFortress2'''s [[{{Delinquent}} Scout]] seems to be a somewhat DarkerAndEdgier, "grown-up" parody of this trope. Even in his mid-twenties, he clearly hasn't matured at all. He's a rough-and-tumble, mischievous baseball fan from Boston in the 60's with a habit of calling others "chuckleheads" and the like, and carries around a baseball bat... which he uses to bash others' head in. [[http://www.teamfortress.com/scoutupdate/force-a-nature.htm Hot dog!]]
* ''VideoGame/WolfensteinTheNewOrder'''s [[NewMeat Private Wyatt]] tends towards this. He's young, idealistic, and [[GoshDangItToHeck almost never swears,]] even in combat. If he [[spoiler: survives the first encounter with [[BigBad Deathshead,]]]] his partner in the resistance is a similarly young Rock'n'Roll enthusiast.
[[folder: Western Animation ]]
* According to WordOfGod, Rocky of ''WesternAnimation/RockyAndBullwinkle'' is an All-American Boy in squirrel form.
* Hank and Dean Venture of ''WesternAnimation/TheVentureBrothers'', at least initially.
* Davey Hansen in ''WesternAnimation/DaveyAndGoliath''.
* Orel Puppington of ''MoralOrel'' is a Deconstruction of this; it's specifically a parody of DaveyAndGoliath
* Ralphie Tennelli from ''TheMagicSchoolBus''.
* Hank Hill of ''WesternAnimation/KingOfTheHill'' thinks Bobby should be one. Bobby's knack for marksmanship and attending meetings of the Order of the Straight Arrow means he fulfills it from time to time.
[[folder: Real Life ]]
* Institutionally invoked by the Boy Scouts of America.
* ''Boys' Life'' magazine is marketed to this demographic, which makes sense as it's published by the aforementioned Boy Scouts.


text:
Shorts sales/foreclosure process?
I also posted this on the Homes forum, but I thought I would ask try here as well. After a year of trying to sell our old home without success, we hired a lawyer to help negotiate a short sale or deed in lieu of foreclosure with our lenders. When we first met with this lawyer, he said it would be highly possible to get our second lender to settle for $5000 instead of the $50000 we owe. He also said he has realtors he works with regularly for short sales. Since we hadn't been able to find a realtor experienced with short sales and had trouble even finding a lawyer to take the time to meet with us, we went ahead and retained him, even though it all sounded a little too easy. That was three months ago, and so far he has done nothing. He said that the forclosure process takes at least six months and that our lenders would not even consider anything until we were at least three months behind on payments. This month we are officially three months behind.
This lawyer also advised us to rent out the house and not tell the property management company we hired what is going on with the house. So now we are contracted with a prop management company and have renters in the house on a 6 month lease. Meanwhile our lenders are still calling us, the lawyer hasn't done anything except send us an itemized bill that shows almost all of the retainer being spent on things like "dicated memo to client" and faxes to the lenders that were never received. It takes several phone calls to get him to call us back, and then he charges us for the time spend talking with us, even though the purpose of the call was to remind him to follow up on things he had not done.
I feel very uncomfortable with this whole situation, not making the payments and having this nice unsuspecting family living in the house not knowing that they could end up getting evicted if we foreclose before their lease is up. I checked the laws in the state and the bank can foreclose in as little as 90 days, which is where we are at. The few times I have spoken with the lawyer since the original meeting, he has assured me that this will take at least 6 months and we will have plenty of time to end the contract with the prop management co. and get the tenants out, but I have to wonder if this is correct.
Does it sound we were taken by an attorney who has no intention of helping us, or is this the normal process? He told us not to talk with the lenders, but I ended up having to the other day because I couldn't get in touch with him to do it for us and now I am nervous I gave away too much information. If anyone has experience and advice on this kind of situation, I would appreciate the input. Thank you.
Hi, this is a too-bad situation. What you really needed to do is first somehow get references on this person, even if it meant going to the local Bar association or somewhere to do it, because it does sound like he may not have done the proper thing. I'm not speaking of the process itself, the sequence, etc., as I certainly don't know anything about it, but the way in which he seems to have gone about things in general is a bit strange, not the least of which would be 'counselling' you to be shifty with the mgmt. co. I do not think it's a 'normal' way to do anything and I'd ask the Association or the local real estate board what to do about it all. You don't want to get worse advice now (from others) on top of bad. Good luck!
Sounds like you hired a scumbag.
Will the second holder settle for $5k on $50k - probably not. They might be wise to since they are likely to get zero, but "banks" and "wise" aren't in the same sentence too often recently. Frankly, the chance of a short sale working with 2 lenders involved is pretty slim.
Will the lender foreclose? Eventually. You lawyer is probably right that it will take a while though.
His advice about renting the house is incredibly unethical. Fortunately, recent laws protect the renters from your unethical behavior. Even if the property is foreclosed on, the new owners are bound by the 6 month lease. The current tenants cannot be evicted as long as they make payments.
At this point, your best bet is to contact the National Foundation of Credit Counseling. That is the real, legit public service group - not one of the infomercial people. They have links on their site for finding a certified counselor in your area. I'm sure they will tell you to drop this bozo lawyer and to start making realistic plans to improve your financial situation.
Here is a link that might be useful: NFCC


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Latest The Lancet Stories
AMSTERDAM, November 4, 2014 /PRNewswire/ -- EBioMedicine brings together worlds of basic research and clinical science in a new, open access journal
King Richard III of England died after suffering multiple blows from weapons, UK researchers have found. Modern forensic analysis of the king's 500 year old remains has identified where the fatal blows probably landed, and what kinds of weapons were likely to have delivered them.
LONDON, September 5, 2014 /PRNewswire/ -- Clinical Microbiology and Infection and New Microbes & New Infections to be published
AMSTERDAM, August 14, 2014 /PRNewswire/ -- Elsevier [http://www.elsevier.com ], a world-leading provider of scientific, technical and medical information products and services,
The Lancet Oncology today publishes a major new Commission examining the challenges to effective cancer control in China, India, and Russia – which together experience 46% of all new cancers worldwide, and account for more than half (52%) of all cancer deaths globally.
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- Forsooth! indeed! originally a parenthetical phrase used in repeating the words of another with more or less contempt or disdain.


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From The Collaborative International Dictionary of English v.0.48 [gcide]:
Envenom \En*ven"om\, verb (used with an object) [imp. & p. p. {Envenomed}; p. pr. & vb. n. {Envenoming}.] [OE. envenimen, F. envenimer; pref. en- (L. in) + F. venin poison. See {Venom}.]
1. To taint or impregnate with venom, or any substance noxious to life; to poison; to render dangerous or deadly by poison, as food, drink, a weapon; as, envenomed meat, wine, or arrow; also, to poison (a person) by impregnating with venom.
Alcides . . . felt the envenomed robe. --Milton.
O, what a world is this, when what is comely Envenoms him that bears it! --Shak.
2. To taint or impregnate with bitterness, malice, or hatred; to imbue as with venom; to imbitter.
The envenomed tongue of calumny. --Smollett.
On the question of slavery opinion has of late years been peculiarly envenomed. --Sir G. C. Lewis.
From WordNet (r) 3.0 (2006) [wn]:
1: cause to be bitter or resentful; "These injustices embittered her even more" [syn: {embitter}, {envenom}, {acerbate}]
2: add poison to; "Her husband poisoned her drink in order to kill her" [syn: {poison}, {envenom}]
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