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Located in Spring Valley, Fairfield Inn by Marriott Spring Valley is close to St. Paul's Episcopal Church and New York Country Club. This hotel is within the vicinity of Rockland County Courthouse and Rockland Community College.
Make yourself at home in one of the 104 air-conditioned rooms featuring refrigerators and microwaves. Complimentary wired and wireless Internet access keeps you connected, and cable programming provides entertainment. Conveniences include desks and coffee/tea makers, and housekeeping is provided daily.
Rec, Spa, Premium Amenities
Take advantage of recreation opportunities including a fitness center and a seasonal outdoor pool.
A complimentary continental breakfast is included.
Business, Other Amenities
Featured amenities include complimentary newspapers in the lobby, dry cleaning/laundry services, and laundry facilities. Free self parking is available onsite.
Are you affiliated with Fairfield Inn Spring Valley Nanuet? Apply to the Hotel WiFi Connect program to take control of your account, improve your hotel's confidence score and ranking.
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Sikh delegation meets Prime Minister Narendra Modi at his residence
A delegation of the Gurudwara visited the Prime Minister at his residence at 7 Lok Kalyan Marg today to offer prasad and blessings
New Delhi: Delhi’s Gurdwara Shri Bala Sahib Ji organised an ‘Akhand paath’ on the occasion of Prime Minister Narendra Modi’s birthday. This ‘Akhand paath’ was attended by thousands of Sikh devotees.
It was perhaps the first of its kind initiative when a Gurdwara organised ‘Akhand paath’ for the Prime Minister of the country. Langer, Heath Camp and Blood donation camp were also organised by the Gurdwara on the occasion.
A delegation of the Gurudwara visited the Prime Minister at his residence at 7 Lok Kalyan Marg today to offer prasad and blessings.
The Sikh delegation honoured the Prime Minister by tying a Pagdi and offering a Siropa. An ardas was also performed for the Prime Minister’s long life and good health.
#WATCH | A delegation of Delhi’s Gurdwara Shri Bala Sahib Ji visited Prime Minister Narendra Modi's residence after the Gurdwara organised an ‘Akhand paath’ which started on 15th September and culminated on 17th September, on his birthday. pic.twitter.com/G2yAi4BAEL
— ANI (@ANI) September 19, 2022
Prime Minister expressed his happiness on meeting the delegation and thanked them for their wishes and prayers for his well-being. He said that he was humbled and deeply touched by their gesture for making him feel a part of the Sikh Community.
He reiterated the commitment of the government for continuously working towards the welfare of the Sikh community.
During the meeting, the delegation also thanked the Prime Minister for the path-breaking initiatives taken by him for the honour and welfare of the Sikh community. They recounted several efforts done by the Prime Minister including declaring 26 December as “Veer Baal Divas”, reopening the Kartarpur Sahib Corridor, removal of GST on langars run by gurdwaras, ensuring that the copies of Guru Granth Sahib reached India from Afghanistan, among others.
For those who worship Durga, Mahalaya rings in the festive flavour. Although this year, Durga Puja is still a month away, eminent personalities have extended regards and greetings on Mahalaya via Twitter
PM Modi launches National Logistics Policy, calls it important step towards making of developed India
The Prime Minister called the policy a solution for many problems and said that it would lead to improvements of "all our systems"

domain: news


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today i have a set of frosty winter theme ATC's to share.You can checkout my tutorial on Colourarte blog for making these.I recently got the Finnabair snowflake paste and i just love the look and feel of it,it almost gives the real snow effect,so i thought of making some winter theme ATC's.
Its nearly impossible to see snow in southern part of India,and this year there is no sign of rain or winter yet even though its going to november end:((.So i thought why not i create some in my art work..thats the good part of art you can create anything you want whether it exist or not doesn't matter.Doesn't this give you the chilly feel??just incase if you are wondering what design i have done let me remind you everything got buried deep under snow,..lol,but you can check out my tutorial if you are curios to see what i have used.
13 Art challenge: Frozen in time
Mixed media place: November challenge

domain: design


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Florida Bar chooses first Black president
1/25/2012, 7:30 a.m.
Attorney Eugene K. Pettis selected by his peers Fort Lauderdale attorney, Eugene K. Pettis, 51, has been chosen as the next president of The Florida Bar. When he is sworn in as president-elect at the Bars annual convention in June 2012, and takes over in 2013, he will become the first Black to hold the position. Its an honor to be recognized by my colleagues and we have 93,000 lawyers in the state that are members of The Florida Bar, he said. Its assuring to know they feel I am worthy to serve as their president and I am proud that we have finally broken the race barrier. The Bar was a voluntary organization in 1907 and became a unified, or mandatory organization in 1951. Either date you chose, its clear that we have waited way too long and ignored quality people of color who were able and worthy to serve but were not allowed. I am glad we are on the brink of removing one of the barriers but there are many others that must follow like gender. Pettis, a co-founder of Haliczer Pettis & Schwamm, attended the University of Florida and then the University of Floridas Levin College of Law. The firm was founded in 1996. When he takes over as president of The Florida Bar, he will represent the countrys second-largest bar with a membership in excess of 95,000 attorneys. I will be working in Tallahassee to advocate for the court system to make sure there is adequate funding, he said. Over the past few years we have seen a shortage of money and a disruption of the court system. My goal will be to work with the legislation and the governor to secure a dedicated source of funding. Pettis says there are other topics that while not specifically listed under the duties of his job, he plans to focus on when he becomes president, including: greater diversity among the leadership of The Florida Bar; mentoring and developing young attorneys; and pushing for a better public education system. In Broward County over 53 percent of our Black boys are not graduating from high school, he said. I grew up in Fort Lauderdale and when I visit the jails I often run into old friends or classmates. When we allow young boys to fail and not complete school, we are providing the perfect pipeline to the prison system. Miami-Dade County is in a similar state. We have to begin having some serious dialogue on how to change these issues. The numbers and statistics come out year after year its time that Black professionals commit themselves to ending this cycle. By D. Kevin McNeir email@example.com

domain: law


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How To Get Tested For Std Chesterfield MO 63005
Truths About Sexually Transferred Illness in Chesterfield MO
Illness which spread out through sexual contact are referred to as “Sexually Sent Illness” or Sexually transmitted diseases. As Everett Koop, MD, Former US General Cosmetic surgeon put it “When you make love with someone, you are having sex with everybody they have had sex with for the last 10 years, and everybody they and their partners have actually made love with for the last ten years.”
Here are some facts about Sexually transmitted diseases:
- Although Sexually transmitted diseases impact males and females, the health issue caused due to STDs might be more serious for women.
- The main causes of Sexually transmitted diseases are bacteria, parasites and infections.
- Chlamydial Infection is the most typical of all bacterial Sexually transmitted diseases and it may cause pelvic inflammatory disease (PID) in ladies.
- Gonorrhea is one of the most frequently reported transmittable illness in the United States.
- The extremely first signs of HIV infection may be flu-like signs and inflamed glands, which might appear within a month or 2. Serious signs might take years to appear.
- People who have been contaminated can make it through for several years with medication to battle the HIV infection.
- Sexually transmitted diseases might trigger cervical and other cancers, pelvic inflammatory disease, chronic liver disease and infertility in women.
The risk of acquiring STD is high amongst youngsters who indulge in sexual activity and increases when an individual has numerous sex partners.
Individuals who are contaminated with STDs are most likely to obtain HIV infection when exposed to the virus through sexual contact than uninfected individuals.
A variety of intervention research studies have actually revealed that detection and treatment of Sexually transmitted diseases may decrease transmission of the HIV virus. There are a variety of sites which use useful info on STDs. You can likewise visit a clinic to get yourself checked for HIV.
Top STD Evaluating Tips in Chesterfield MO
Sexually Transmitted Disease screening is critical for men and females who are active sexually. The most common sexually transmitted diseases will be screened by health care service providers. Some of the most common ones include Chlamydia, HIV, Gonorrhea and herpes; the list goes on.
When it concerns herpes, it is hard to identify since the indications or signs are mainly the only evidence; and may appear later on. Syphilis screening is usually suggested to women who are expectant. The following is a breakdown of the elements and tips while testing for SEXUALLY TRANSMITTED DISEASE.
There is Sexually Transmitted Disease screening for blood diseases like HIV and Syphilis. Testing the other sexually transmitted conditions will include taking various samples from impacted areas of the body.
Health experts recommend males and females to opt for STD testing once a year. This will be to inspect for conditions pointed out above consisting of the notorious HIV. Considering that it is hard to understand whether Herpes is present, those with typical signs for the condition should take action prior to the illness intensifies.
Your general physician or healthcare company should be in position to offer STD testing.
Like the time interval that is pegged to each sexually transferred illness concerning screening. For example, HIV screening requires you to do it again after 3 months and again to totally establish the real outcomes. Some Sexually transmitted diseases like Chlamydia require a week to be discovered after sexual relations.
Apart from blood samples, Sexually Transmitted Disease screening as discussed above will involve taking swabs and for instance in males, swabs are drawn from the anus or urethra (remembering sexual preference).
One week suffices to know the outcomes of the majority of tests. If those results are positive, there are treatments/cures offered for a lot of STIs. However, those with the HIV infection might only look forward to managing their condition due to the fact that a treatment is still elusive.
With STDs, avoidance is the sure way to win.
Sexually Transmitted Disease Screening at Home Has Advantages in 63005 Missouri
Medical and health personnel have actually been worrying the significance of getting evaluated for stds. People are now getting educated about std screening and how it can prevent them from spreading diseases or getting them. Below are some benefits of using std home testing.
Privacy in your home in Chesterfield Missouri
The sexually transmitted disease house screening will provide you a customer number and account simply for you. Personal privacy likewise keeps you out of the doctor’s workplace or those totally free centers for sexually transmitted disease screening where everyone will seem to evaluate you.
Convenience of Testing in your home Chesterfield MO
There are std testing sets that can be finished in your house at your own threats. Totally practical for home screening, doing it yourself implies doing all the work and research to make sure the test is done correctly. If you do have a std according to the test, be sure to contact your physician for a visit.
Online Forums for STDs Chesterfield 63005
If you are in need of talking to someone about sexually transmitted disease screening or about where to find totally free clinics for std screening, go online to online forums. Individuals love to talk to others in online forums and lots of will be pleased to provide out details they currently have or refer you to resources they might have access to.
Guarantee Sexually Transmitted Disease Screening Is Done Chesterfield MO
There are cases that have been discussed on the getSTDtested.com site where patients went to their doctor for std testing and were rejected. Some doctors have their own view on screening for such diseases and may feel there is not a requirement, and regrettably it can take place. When you do the std house testing, you will have the opportunity of ensuring it gets done and nobody can refuse you.
Do not let a disease scare you away from getting evaluated. Now that you can gain from sexually transmitted disease screening in your home, you should not be fretted that you may be living with something that could make you self mindful.Where Do You Get Tested For Stds Chesterfield MO 63005
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domain: medical


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National POW/MIA Recognition Day is observed across the United States on the third Friday of September each year. Many Americans take the time to remember those who were prisoners of war (POW) and those who are missing in action (MIA), as well as their families.
National POW/MIA Awareness Day gives many Americans across the United States the time to pause to remember the sacrifices and service of those who were prisoners of war (POW), as well as those who are missing in action (MIA), and their families. All military installations fly the National League of Families’ POW/MIA flag, which symbolizes the nation’s remembrance of those who were imprisoned while serving in conflicts and those who remain missing.
The awareness color for POW/MIA Remembrance is Black.
Personalized Cause supports National POW/MIA Remembrance Day with:

domain: sociology


While it all started out fairly close, Port of Parnu soon managed to take control, as they rocketed ahead in the final minutes of the first quarter with an 8-0 run, giving them a 20-11 advantage.
The margin increased to a game-high 12 points in the second quarter, but the Jurmala side were far from breaking down completely.
After the break, the Latvians managed to cut into the deficit very quickly, and nothing but three points separated the two clubs after three quarters.
Jurmala/Fenikss completed the comeback in the final quarter, as they tied it up on the 33rd minute and then jumped ahead. The final minutes were nerve-wrecking for both sides, as neither team managed to get anything to fall.
That was in favor of the Latvian team, of course, as they eventually got the victory.
Arturs Strelnieks was the leading scorer for Jurmala, as he tallied 14 points and 5 rebounds. Meanwhile, Jekabps Rozitis added 12, while Ugis Pinete chipped in with 9 points, 7 rebounds and 5 assists.
On the other side, it was Vaidotas Volkus with the best offensive output, as he scored 16 points. Saimon Sutt, Reinar Hallik and Norman Kabin added 9 points apiece.


If you're looking to get your feet wet in the road racing scene, or merely want a bike that does it all for your weekend rides, look no further than the Tarmac Comp.
For the construction, we utilized advanced aerospace composite optimization software to revolutionize the construction and layup of our new FACT 10r carbon. It's the most advanced layup schedule we've ever made—the shape, schedule, and material optimization allowed us to shed nearly 20% of the frame weight—the perfect recipe for your next hill climb PR.
We started by further improving our Rider-First Engineered™ technology to ensure that the new Tarmac is stiff and compliant in exactly the right places, all while shedding some serious weight. With this revamp, we also updated the geometry—basing it on countless Retül data points and professional rider input. This enabled us to develop a performance road geometry that perfects the combination of a responsive front-end and short wheelbase, delivering instantaneous response and optimal power transfer.
And while stiffness aids in the aforementioned, compliance must also be utilized for an optimal ride quality. That's why we designed a seatpost that builds compliance into the upper 120mm where clamping doesn't happen, dropped the seatstays, and altered the seat tube shape. We also added tyre clearance up to 30mm, which translates to a 28mm Turbo Cotton on a Roval SLX 24 rim. This allows lower pressures for decreased rolling resistance, increased traction, and more comfort. Altogether, these additions still have the Tarmac riding like a true race-machine, but they just take a bit of the sting out of those road imperfections. You'll thank us on your next long ride.
And while this is all great, we know that aerodynamics are the most important thing we can do to make you faster. Both our Bora-Hansgrohe and Quick-Step Floors Pro Tour riders, after all, are demanding aero improvements on every bike. With this, the aero goal was to discover where we could essentially "add aero for free," by not taking anything away from the hallmarks of the Tarmac design. During the six-month iterative process, three areas were discovered where we could do this—a new fork shape, dropped seatstays with aero tubes, and a D-shaped seatpost and seat tube. The result? A bike that's approximately 45 seconds faster over 40km compared to other lightweight bikes in the same category.
This Tarmac comes ready to race out of the box, featuring a crisp Shimano Ultegra 8000 mechanical groupset, resplendent DT R460 wheels, and smooth-as-silk 26mm Turbo Pro tyres.
- Featuring our Rider-First Engineered™ design that ensures every frame size has the same legendary climbing responsiveness and descending prowess you'd expect from a Tarmac. The FACT 10r frame is the perfect blend of value and performance.
- The 11-speed Shimano Ultegra 8000 shifting is extremely reliable, lightweight, and precise.
- DT R460 wheels feature reliable, sealed cartridge hubs, and 20 front and 24 rear spokes that keeps weight low and durability high.
- Tech Specs
Frame Tarmac SL6, FACT 10r carbon, Rider-First Engineered™, OSBB, clean routing, internally integrated seat clamp, 130mm rear spacing, direct-mount rim brake Fork FACT 10r carbon, direct-mount rim brake Stem Specialized, 3D-forged alloy, 4-bolt, 7-degree rise Handlebars Specialized Shallow Drop, 6061, 70x125mm, 31.8mm clamp Tape S-Wrap Roubaix w/ sticky gel Front Brake Shimano Ultegra R8010-F, direct-mount Rear Brake Shimano Ultegra R8010-F, direct-mount Front Derailleur Shimano Ultegra R8000, braze-on Rear Derailleur Shimano Ultegra R8000-GS, medium cage Shifters Shimano Ultegra R8000 Bottom Bracket Praxis, BB30 for Shimano Cassette Shimano Ultegra R8000, 11-speed, 11-30t Chain Shimano Ultegra, 11-speed Crankset Shimano Ultegra R8000, HollowTech 2, 11-speed Front Wheel DT R460, sealed cartridge hubs, 14g spokes, 20h Rear Wheel DT R460, sealed cartridge hubs, 14g spokes, 20h Front Tyre Turbo Pro, 60 TPI, folding bead, BlackBelt protection, 700x26mm Rear Tyre Turbo Pro, 60 TPI, folding bead, BlackBelt protection, 700x26mm Saddle Toupe Comp Gel, 143mm, hollow Cro-Mo rails, Adaptive Edge Seatpost S-Works FACT Carbon Tarmac seatpost, 20mm offset
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UK orders are sent using Royal Mail Tracked 48, Royal Mail Recorded or Royal Mail letter post for small, light orders and Parcelforce for heavier, bulky orders. Please allow approx 2-3 days for delivery after the order has been despatched. Large and heavy items, e.g turbo trainers and work stands, will be sent by Parcelforce (UK Mainland only). Overseas orders are sent using Royal Mail International Signed For service and Royal Mail Airsure service (where permitting). Heavier items e.g. bicycles are sent using Parcelforce Worldwide. Delivery times will vary depending on destination, customs clearance, local postal service etc. Please allow up to 14 working days for delivery. Postage costs include Insurance for the value of the order. Bicycles sent within the UK are sent by Parcelforce 48 Hour Service.


An Unpopular Opinion But... #4 Submitted by Tom O'Hare on 10/13/2018 at 10:46 AM
The brand extension is flailing.
In this week’s column where I look at what I believe to be unpopular opinions among wrestling fans, I am going to dissect why I believe the brand extension is currently floundering & potentially doing more harm than good to the WWE weekly television shows.
On the face of it, the brand extension should work wonders for World Wrestling Entertainment & all of its employees. A lighter schedule, television time for those who wouldn’t otherwise receive it & talent being less over saturated to the point of boredom for fans. The points mentioned above are the positives of the brand extension (or so I believe) however they are outweighed by the negatives which in turn is having a detrimental effect on the WWE product.
WWE would like you to believe that Raw & Smackdown are on a level playing field & are equal in terms of their presentation, story telling, talents & overall viewing experience.
While there is no doubt Smackdown has very often been a better show as far as entertainment is concerned, it’s abundantly clear the WWE still perceives it as the so called lesser show.
The obvious starting point here is the small matter of Smackdown having 60 minute less running time to it on a weekly basis. Fans are therefore conditioned to believe that Raw is more integral given the fact it appears on television for a longer period of time. I am in no way suggesting Smackdown should be 3 hours long by the way, more so that Raw & Smackdown should be equals in how they are presented to us as viewers.
The second point, most things of significance happen on Raw without any real story line explanation given as to why not Smackdown. Let me elaborate; When Goldberg made his long awaited return to the ring after 12 years away from active completion which brand was it he would go to? Sure, he was feuding with Brock Lesnar ( a Raw superstar) but the WWE were again guilty of conditioning fans to believe Raw was the A-Show & a Goldberg like superstar was only worthy of appearing on said show. Ronda Rousey is another example as well as the recently returned Shawn Michaels. John Cena too; an initial Smackdown superstar before the WWE, without explanation, decided he was going to be a free agent & could appear on both programmes. By making Raw appear a higher priority it diminishes the efforts of those on Smackdown.
The third point & perhaps most pertinent one is the lack of pay-per-view main events given to Smackdown talents, most notably WWE Champion A.J Styles. This I believe is not something that is going to change any time soon & thus reaffirming my stance that Raw is a bigger priority to the WWE higher ups than Smackdown. Even the Royal Rumble, as historic as the inaugural woman's bout was, concluded with the lasting image of Asuka’s victory & Ronda Rousey’s debut. Both of whom of course were Raw superstars at the time. Shinsuke Nakamura, a Smackdown star, had to settle for his victory being fourth to last on the card.
The fourth & final point I’m going to write about is the constant presence of one Corey Graves on both shows week in & week out. Earlier in the article I spoke about over saturation of talent & how this can lead fans to feeling that the individual in question has gotten stale. Corey Graves, talented as he is, does not need to appear on both programmes on consecutive nights in consecutive weeks. The brand extension, as WWE would have you believe, offer talent who are exclusive to their brand meaning you as a viewer have to watch whichever show they are on if you are a fan of their’s. With Graves constantly appearing on both Raw & Smackdown it becomes incredibly difficult for us as fans not to get complacement with him as well as being able to identify with his character as “The Raw Guy” or “The Smackdown Guy”. Instead he’s “The guy who appears on both shows & sometimes like heels & sometimes likes faces”. A mistake easily solved in my opinion but one which WWE persists in making.
In closing, I DON’T believe WWE ARE making an error in their idea of the brand extension BUT rather their presentation of it.
My name is Tom O’Hare & this has been my unpopular opinion. Thank you for reading.
*If you’d like to find me on social media I can be located via numerous platforms:


DINNINGTON were denied victory by a last minute try but then saved from defeat by the width of the upright.
After a three week break, Dinnington visited fifth-placed York hoping to continue the good form that they ended 2012 with.
However, York had other ideas and started the game at a furious pace which forced Dinnington back towards their own line.
In the opening minute, Dinnington gave away a string of penalties but good defending prevented York claiming the try they were clearly looking for.
Instead, York had to accept three points from a penalty kick, but three minutes later they did get over the line after spinning the ball along their backs and scoring in the corner.
Dinnington got themselves into the game by driving at the home side with their forwards and they enjoyed a period of pressure in the home half resulting in three points from a penalty kick by Jonny West.
York came straight back but were kept out by some excellent defending and then play switched from end to end.
On their next visit to the York half, another penalty from West cut the deficit to two points but 10 minutes from half time York restored their five point advantage with a penalty of their own.
As both sides battled to gain the upper hand before half-time, discipline began to slip and both sides had a player sent to the bin; firstly, York and then Dinnington a couple of minutes later.
Dinnington made a much better start to the second half, but with only a few minutes played had another player binned, resulting in them having to play two players light for a short time.
However, it was hard to tell as they took the game to York and enjoyed the majority of possession.
Back to full strength, Dinnington began to get on top and just short of the mid-point of the half took the lead when West threw a dummy pass that fooled everyone giving him an easy walk in under the posts.
His conversion put Dinnington 13-11 ahead and another penalty five minutes later opened the gap to five points.
Dinnington then set about trying to defend their lead by keeping the ball close to the forwards but York still had chances and had the home number eight off-loaded more instead of running into tacklers, York could easily have run in several tries.
The visitors defended well, often close to their own line and the more they kept the home side out the more their confidence grew.
In the final minutes West attempted a drop-goal to stretch the lead to more than one score but his effort went just wide.
Two minutes from time, York knocked-on giving Dinnington a scrum inside their own 22.
Dinnington had not struggled at the scrum all game but York turned the ball over and quickly moved it along their backs, scoring in the corner.
Dinnington were deflated at having victory snatched away from them but they were more concerned as the conversion attempt headed towards the posts.
There was a big sigh of relief when it hit the upright and rebounded clear. That was the last action of the game.
Tomorrow, Dinnington are at home to Barnsley.


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When: Tue., June 21, 6:30 p.m. 2016
Cha Gio is the Vietnamese version of the egg roll, served with a delicious dipping sauce on a bed of lettuce. In this class participants will make and then enjoy these egg rolls. Students leave with recipes and confidence to make these at home. For ages 18 and up.

domain: food


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The stress hormone cortisol (CORT) is slowly incorporated into the growing hair shaft of humans, nonhuman primates, and other mammals. We developed and validated a method for CORT extraction and analysis from rhesus monkey hair and subsequently adapted this method for use with human scalp hair. In contrast to CORT "point samples" obtained from plasma or saliva, hair CORT provides an integrated measure of hypothalamic-pituitary-adrenocortical (HPA) system activity, and thus physiological stress, during the period of hormone incorporation. Because human scalp hair grows at an average rate of 1 cm/month, CORT levels obtained from hair segments several cm in length can potentially serve as a biomarker of stress experienced over a number of months.
In our method, each hair sample is first washed twice in isopropanol to remove any CORT from the outside of the hair shaft that has been deposited from sweat or sebum. After drying, the sample is ground to a fine powder to break up the hair's protein matrix and increase the surface area for extraction. CORT from the interior of the hair shaft is extracted into methanol, the methanol is evaporated, and the extract is reconstituted in assay buffer. Extracted CORT, along with standards and quality controls, is then analyzed by means of a sensitive and specific commercially available enzyme immunoassay (EIA) kit. Readout from the EIA is converted to pg CORT per mg powdered hair weight. This method has been used in our laboratory to analyze hair CORT in humans, several species of macaque monkeys, marmosets, dogs, and polar bears. Many studies both from our lab and from other research groups have demonstrated the broad applicability of hair CORT for assessing chronic stress exposure in natural as well as laboratory settings.
17 Related JoVE Articles!
Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis
Institutions: National Jewish Health, University of Colorado Denver.
Metabolomics is an emerging field which enables profiling of samples from living organisms in order to obtain insight into biological processes. A vital aspect of metabolomics is sample preparation whereby inconsistent techniques generate unreliable results. This technique encompasses protein precipitation, liquid-liquid extraction, and solid-phase extraction as a means of fractionating metabolites into four distinct classes. Improved enrichment of low abundance molecules with a resulting increase in sensitivity is obtained, and ultimately results in more confident identification of molecules. This technique has been applied to plasma, bronchoalveolar lavage fluid, and cerebrospinal fluid samples with volumes as low as 50 µl. Samples can be used for multiple downstream applications; for example, the pellet resulting from protein precipitation can be stored for later analysis. The supernatant from that step undergoes liquid-liquid extraction using water and strong organic solvent to separate the hydrophilic and hydrophobic compounds. Once fractionated, the hydrophilic layer can be processed for later analysis or discarded if not needed. The hydrophobic fraction is further treated with a series of solvents during three solid-phase extraction steps to separate it into fatty acids, neutral lipids, and phospholipids. This allows the technician the flexibility to choose which class of compounds is preferred for analysis. It also aids in more reliable metabolite identification since some knowledge of chemical class exists.
Bioengineering, Issue 89, plasma, chemistry techniques, analytical, solid phase extraction, mass spectrometry, metabolomics, fluids and secretions, profiling, small molecules, lipids, liquid chromatography, liquid-liquid extraction, cerebrospinal fluid, bronchoalveolar lavage fluid
Biochemical Measurement of Neonatal Hypoxia
Institutions: Loma Linda University, Loma Linda University.
Neonatal hypoxia ischemia is characterized by inadequate blood perfusion of a tissue or a systemic lack of oxygen. This condition is thought to cause/exacerbate well documented neonatal disorders including neurological impairment 1-3
. Decreased adenosine triphosphate production occurs due to a lack of oxidative phosphorylation. To compensate for this energy deprived state molecules containing high energy phosphate bonds are degraded 2
. This leads to increased levels of adenosine which is subsequently degraded to inosine, hypoxanthine, xanthine, and finally to uric acid. The final two steps in this degradation process are performed by xanthine oxidoreductase. This enzyme exists in the form of xanthine dehydrogenase under normoxic conditions but is converted to xanthine oxidase (XO) under hypoxia-reperfusion circumstances 4, 5
. Unlike xanthine dehydrogenase, XO generates hydrogen peroxide as a byproduct of purine degradation 4, 6
. This hydrogen peroxide in combination with other reactive oxygen species (ROS) produced during hypoxia, oxidizes uric acid to form allantoin and reacts with lipid membranes to generate malondialdehyde (MDA) 7-9
. Most mammals, humans exempted, possess the enzyme uricase, which converts uric acid to allantoin. In humans, however, allantoin can only be formed by ROS-mediated oxidation of uric acid. Because of this, allantoin is considered to be a marker of oxidative stress in humans, but not in the mammals that have uricase.
We describe methods employing high pressure liquid chromatography (HPLC) and gas chromatography mass spectrometry (GCMS) to measure biochemical markers of neonatal hypoxia ischemia. Human blood is used for most tests. Animal blood may also be used while recognizing the potential for uricase-generated allantoin
. Purine metabolites were linked to hypoxia as early as 1963 and the reliability of hypoxanthine, xanthine, and uric acid as biochemical indicators of neonatal hypoxia was validated by several investigators 10-13
. The HPLC method used for the quantification of purine compounds is fast, reliable, and reproducible. The GC/MS method used for the quantification of allantoin, a relatively new marker of oxidative stress, was adapted from Gruber et al 7
. This method avoids certain artifacts and requires low volumes of sample. Methods used for synthesis of MMDA were described elsewhere 14, 15
. GC/MS based quantification of MDA was adapted from Paroni et al
. and Cighetti et al
. 16, 17
. Xanthine oxidase activity was measured by HPLC by quantifying the conversion of pterin to isoxanthopterin 18
. This approach proved to be sufficiently sensitive and reproducible.
Medicine, Issue 54, hypoxia, Ischemia, Neonate, Hypoxanthine, Xanthine, Uric Acid, Allantoin, Xanthine Oxidase, Malondialdehyde
A High-throughput Method for Measurement of Glomerular Filtration Rate in Conscious Mice
Institutions: University of California, San Diego , San Diego VA Healthcare System.
The measurement of glomerular filtration rate (GFR) is the gold standard in kidney function assessment. Currently, investigators determine GFR by measuring the level of the endogenous biomarker creatinine or exogenously applied radioactive labeled inulin (3
H or 14
C). Creatinine has the substantial drawback that proximal tubular secretion accounts for ~50% of total renal creatinine excretion and therefore creatinine is not a reliable GFR marker. Depending on the experiment performed, inulin clearance can be determined by an intravenous single bolus injection or continuous infusion (intravenous or osmotic minipump). Both approaches require the collection of plasma or plasma and urine, respectively. Other drawbacks of radioactive labeled inulin include usage of isotopes, time consuming surgical preparation of the animals, and the requirement of a terminal experiment. Here we describe a method which uses a single bolus injection of fluorescein isothiocyanate-(FITC) labeled inulin and the measurement of its fluorescence in 1-2 μl of diluted plasma. By applying a two-compartment model, with 8 blood collections per mouse, it is possible to measure GFR in up to 24 mice per day using a special work-flow protocol. This method only requires brief isoflurane anesthesia with all the blood samples being collected in a non-restrained and awake mouse. Another advantage is that it is possible to follow mice over a period of several months and treatments (i.e.
doing paired experiments with dietary changes or drug applications). We hope that this technique of measuring GFR is useful to other investigators studying mouse kidney function and will replace less accurate methods of estimating kidney function, such as plasma creatinine and blood urea nitrogen.
Medicine, Issue 75, Anatomy, Physiology, Biomedical Engineering, Molecular Biology, Nephrology, Kidney Function Tests, Glomerular filtration rate, rats, mice, conscious, creatinine, inulin, Jaffe, hypertension, HPLC, animal model
Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo
Institutions: INSERM, Hôpitaux Universitaires de Strasbourg, Université de Strasbourg.
Metallic implants, especially titanium implants, are widely used in clinical applications. Tissue in-growth and integration to these implants in the tissues are important parameters for successful clinical outcomes. In order to improve tissue integration, porous metallic implants have being developed. Open porosity of metallic foams is very advantageous, since the pore areas can be functionalized without compromising the mechanical properties of the whole structure. Here we describe such modifications using porous titanium implants based on titanium microbeads. By using inherent physical properties such as hydrophobicity of titanium, it is possible to obtain hydrophobic pore gradients within microbead based metallic implants and at the same time to have a basement membrane mimic based on hydrophilic, natural polymers. 3D pore gradients are formed by synthetic polymers such as Poly-L-lactic acid (PLLA) by freeze-extraction method. 2D nanofibrillar surfaces are formed by using collagen/alginate followed by a crosslinking step with a natural crosslinker (genipin). This nanofibrillar film was built up by layer by layer (LbL) deposition method of the two oppositely charged molecules, collagen and alginate. Finally, an implant where different areas can accommodate different cell types, as this is necessary for many multicellular tissues, can be obtained. By, this way cellular movement in different directions by different cell types can be controlled. Such a system is described for the specific case of trachea regeneration, but it can be modified for other target organs. Analysis of cell migration and the possible methods for creating different pore gradients are elaborated. The next step in the analysis of such implants is their characterization after implantation. However, histological analysis of metallic implants is a long and cumbersome process, thus for monitoring host reaction to metallic implants in vivo
an alternative method based on monitoring CGA and different blood proteins is also described. These methods can be used for developing in vitro
custom-made migration and colonization tests and also be used for analysis of functionalized metallic implants in vivo
Biomedical Engineering, Issue 77, Bioengineering, Medicine, Anatomy, Physiology, Biophysics, Cellular Biology, Molecular Biology, Materials Science, Biomedical and Dental Materials, Composite Materials, Metals and Metallic Materials, Engineering (General), Titanium, pore gradient, implant, in vivo, blood analysis, freeze-extraction, foams, implants, transplantation, clinical applications
Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
Institutions: University of Maine.
Localization-based super resolution microscopy can be applied to obtain a spatial map (image) of the distribution of individual fluorescently labeled single molecules within a sample with a spatial resolution of tens of nanometers. Using either photoactivatable (PAFP) or photoswitchable (PSFP) fluorescent proteins fused to proteins of interest, or organic dyes conjugated to antibodies or other molecules of interest, fluorescence photoactivation localization microscopy (FPALM) can simultaneously image multiple species of molecules within single cells. By using the following approach, populations of large numbers (thousands to hundreds of thousands) of individual molecules are imaged in single cells and localized with a precision of ~10-30 nm. Data obtained can be applied to understanding the nanoscale spatial distributions of multiple protein types within a cell. One primary advantage of this technique is the dramatic increase in spatial resolution: while diffraction limits resolution to ~200-250 nm in conventional light microscopy, FPALM can image length scales more than an order of magnitude smaller. As many biological hypotheses concern the spatial relationships among different biomolecules, the improved resolution of FPALM can provide insight into questions of cellular organization which have previously been inaccessible to conventional fluorescence microscopy. In addition to detailing the methods for sample preparation and data acquisition, we here describe the optical setup for FPALM. One additional consideration for researchers wishing to do super-resolution microscopy is cost: in-house setups are significantly cheaper than most commercially available imaging machines. Limitations of this technique include the need for optimizing the labeling of molecules of interest within cell samples, and the need for post-processing software to visualize results. We here describe the use of PAFP and PSFP expression to image two protein species in fixed cells. Extension of the technique to living cells is also described.
Basic Protocol, Issue 82, Microscopy, Super-resolution imaging, Multicolor, single molecule, FPALM, Localization microscopy, fluorescent proteins
A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
Institutions: University of California, San Francisco - UCSF, Buck Institute for Age Research, Purdue University.
Glycans are an important class of post-translational modifications. Typically found on secreted and extracellular molecules, glycan structures signal the internal status of the cell. Glycans on tumor cells tend to have abundant sialic acid and fucose moieties. We propose that these cancer-associated glycan variants be exploited for biomarker development aimed at diagnosing early-stage disease. Accordingly, we developed a mass spectrometry-based workflow that incorporates chromatography on affinity matrices formed from lectins, proteins that bind specific glycan structures. The lectins Sambucus nigra (SNA) and Aleuria aurantia (AAL), which bind sialic acid and fucose, respectively, were covalently coupled to POROS beads (Applied Biosystems) and packed into PEEK columns for high pressure liquid chromatography (HPLC). Briefly, plasma was depleted of the fourteen most abundant proteins using a multiple affinity removal system (MARS-14; Agilent). Depleted plasma was trypsin-digested and separated into flow-through and bound fractions by SNA or AAL HPLC. The fractions were treated with PNGaseF to remove N-linked glycans, and analyzed by LC-MS/MS on a QStar Elite. Data were analyzed using Mascot software. The experimental design included positive controls—fucosylated and sialylated human lactoferrin glycopeptides—and negative controls—high mannose glycopeptides from Saccharomyces cerevisiae—that were used to monitor the specificity of lectin capture. Key features of this workflow include the reproducibility derived from the HPLC format, the positive identification of the captured and PNGaseF-treated glycopeptides from their deamidated Asn-Xxx-Ser/Thr motifs, and quality assessment using glycoprotein standards. Protocol optimization also included determining the appropriate ratio of starting material to column capacity, identifying the most efficient capture and elution buffers, and monitoring the PNGaseF-treatment to ensure full deglycosylation. Future directions include using this workflow to perform mass spectrometry-based discovery experiments on plasma from breast cancer patients and control individuals.
Basic Protocols, Issue 32, Lectins, chromatography, glycopeptides, glycoproteins, biomarker discovery
A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Institutions: University of Sydney, Monash University.
Vitamin C (ascorbate) plays numerous important roles in cellular metabolism, many of which have only come to light in recent years. For instance, within the brain, ascorbate acts in a neuroprotective and neuromodulatory manner that involves ascorbate cycling between neurons and vicinal astrocytes - a relationship that appears to be crucial for brain ascorbate homeostasis. Additionally, emerging evidence strongly suggests that ascorbate has a greatly expanded role in regulating cellular and systemic iron metabolism than is classically recognized. The increasing recognition of the integral role of ascorbate in normal and deregulated cellular and organismal physiology demands a range of medium-throughput and high-sensitivity analytic techniques that can be executed without the need for highly expensive specialist equipment. Here we provide explicit instructions for a medium-throughput, specific and relatively inexpensive microplate assay for the determination of both intra- and extracellular ascorbate in cell culture.
Biochemistry, Issue 86, Vitamin C, Ascorbate, Cell swelling, Glutamate, Microplate assay, Astrocytes
A Strategy for Sensitive, Large Scale Quantitative Metabolomics
Institutions: Cornell University, Cornell University.
Metabolite profiling has been a valuable asset in the study of metabolism in health and disease. However, current platforms have different limiting factors, such as labor intensive sample preparations, low detection limits, slow scan speeds, intensive method optimization for each metabolite, and the inability to measure both positively and negatively charged ions in single experiments. Therefore, a novel metabolomics protocol could advance metabolomics studies. Amide-based hydrophilic chromatography enables polar metabolite analysis without any chemical derivatization. High resolution MS using the Q-Exactive (QE-MS) has improved ion optics, increased scan speeds (256 msec at resolution 70,000), and has the capability of carrying out positive/negative switching. Using a cold methanol extraction strategy, and coupling an amide column with QE-MS enables robust detection of 168 targeted polar metabolites and thousands of additional features simultaneously. Data processing is carried out with commercially available software in a highly efficient way, and unknown features extracted from the mass spectra can be queried in databases.
Chemistry, Issue 87, high-resolution mass spectrometry, metabolomics, positive/negative switching, low mass calibration, Orbitrap
Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays
Institutions: The University of Mississippi.
Much of the nutrient cycling and carbon processing in natural environments occurs through the activity of extracellular enzymes released by microorganisms. Thus, measurement of the activity of these extracellular enzymes can give insights into the rates of ecosystem level processes, such as organic matter decomposition or nitrogen and phosphorus mineralization. Assays of extracellular enzyme activity in environmental samples typically involve exposing the samples to artificial colorimetric or fluorometric substrates and tracking the rate of substrate hydrolysis. Here we describe microplate based methods for these procedures that allow the analysis of large numbers of samples within a short time frame. Samples are allowed to react with artificial substrates within 96-well microplates or deep well microplate blocks, and enzyme activity is subsequently determined by absorption or fluorescence of the resulting end product using a typical microplate reader or fluorometer. Such high throughput procedures not only facilitate comparisons between spatially separate sites or ecosystems, but also substantially reduce the cost of such assays by reducing overall reagent volumes needed per sample.
Environmental Sciences, Issue 80, Environmental Monitoring, Ecological and Environmental Processes, Environmental Microbiology, Ecology, extracellular enzymes, freshwater microbiology, soil microbiology, microbial activity, enzyme activity
Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development
Institutions: Consiglio Nazionale delle Ricerche.
The involvement of free radicals in life sciences has constantly increased with time and has been connected to several physiological and pathological processes. This subject embraces diverse scientific areas, spanning from physical, biological and bioorganic chemistry to biology and medicine, with applications to the amelioration of quality of life, health and aging. Multidisciplinary skills are required for the full investigation of the many facets of radical processes in the biological environment and chemical knowledge plays a crucial role in unveiling basic processes and mechanisms. We developed a chemical biology approach able to connect free radical chemical reactivity with biological processes, providing information on the mechanistic pathways and products. The core of this approach is the design of biomimetic models to study biomolecule behavior (lipids, nucleic acids and proteins) in aqueous systems, obtaining insights of the reaction pathways as well as building up molecular libraries of the free radical reaction products. This context can be successfully used for biomarker discovery and examples are provided with two classes of compounds: mono-trans isomers of cholesteryl esters, which are synthesized and used as references for detection in human plasma, and purine 5',8-cyclo-2'-deoxyribonucleosides, prepared and used as reference in the protocol for detection of such lesions in DNA samples, after ionizing radiations or obtained from different health conditions.
Chemistry, Issue 74, Biochemistry, Chemical Engineering, Chemical Biology, chemical analysis techniques, chemistry (general), life sciences, radiation effects (biological, animal and plant), biomarker, biomimetic chemistry, free radicals, trans lipids, cyclopurine lesions, DNA, chromatography, spectroscopy, synthesis
High-throughput, Automated Extraction of DNA and RNA from Clinical Samples using TruTip Technology on Common Liquid Handling Robots
Institutions: Akonni Biosystems, Inc., Akonni Biosystems, Inc., Akonni Biosystems, Inc., Akonni Biosystems, Inc..
TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. The geometry of the monolith can be adapted for specific pipette tips ranging in volume from 1.0 to 5.0 ml. The large porosity of the monolith enables viscous or complex samples to readily pass through it with minimal fluidic backpressure. Bi-directional flow maximizes residence time between the monolith and sample, and enables large sample volumes to be processed within a single TruTip. The fundamental steps, irrespective of sample volume or TruTip geometry, include cell lysis, nucleic acid binding to the inner pores of the TruTip monolith, washing away unbound sample components and lysis buffers, and eluting purified and concentrated nucleic acids into an appropriate buffer. The attributes and adaptability of TruTip are demonstrated in three automated clinical sample processing protocols using an Eppendorf epMotion 5070, Hamilton STAR and STARplus liquid handling robots, including RNA isolation from nasopharyngeal aspirate, genomic DNA isolation from whole blood, and fetal DNA extraction and enrichment from large volumes of maternal plasma (respectively).
Genetics, Issue 76, Bioengineering, Biomedical Engineering, Molecular Biology, Automation, Laboratory, Clinical Laboratory Techniques, Molecular Diagnostic Techniques, Analytic Sample Preparation Methods, Clinical Laboratory Techniques, Molecular Diagnostic Techniques, Genetic Techniques, Molecular Diagnostic Techniques, Automation, Laboratory, Chemistry, Clinical, DNA/RNA extraction, automation, nucleic acid isolation, sample preparation, nasopharyngeal aspirate, blood, plasma, high-throughput, sequencing
Real-time Analyses of Retinol Transport by the Membrane Receptor of Plasma Retinol Binding Protein
Institutions: University of California, Los Angeles .
Vitamin A is essential for vision and the growth/differentiation of almost all human organs. Plasma retinol binding protein (RBP) is the principle and specific carrier of vitamin A in the blood. Here we describe an optimized technique to produce and purify holo-RBP and two real-time monitoring techniques to study the transport of vitamin A by the high-affinity RBP receptor STRA6. The first technique makes it possible to produce a large quantity of high quality holo-RBP (100%-loaded with retinol) for vitamin A transport assays. High quality RBP is essential for functional assays because misfolded RBP releases vitamin A readily and bacterial contamination in RBP preparation can cause artifacts. Real-time monitoring techniques like electrophysiology have made critical contributions to the studies of membrane transport. The RBP receptor-mediated retinol transport has not been analyzed in real time until recently. The second technique described here is the real-time analysis of STRA6-catalyzed retinol release or loading. The third technique is real-time analysis of STRA6-catalyzed retinol transport from holo-RBP to cellular retinol binding protein I (CRBP-I). These techniques provide high sensitivity and resolution in revealing RBP receptor's vitamin A uptake mechanism.
Biochemistry, Issue 71, Molecular Biology, Genetics, Cellular Biology, Molecular Biology, Anatomy, Physiology, Ophthalmology, Proteomics, Proteins, Membrane Transport Proteins, Vitamin A, retinoid, RBP complex, membrane transport, membrane receptor, STRA6, retinol binding protein
High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease
Institutions: University of Michigan .
Unbiased discovery proteomics strategies have the potential to identify large numbers of novel biomarkers that can improve diagnostic and prognostic testing in a clinical setting and may help guide therapeutic interventions. When large numbers of candidate proteins are identified, it may be difficult to validate candidate biomarkers in a timely and efficient fashion from patient plasma samples that are event-driven, of finite volume and irreplaceable, such as at the onset of acute graft-versus-host disease (GVHD), a potentially life-threatening complication of allogeneic hematopoietic stem cell transplantation (HSCT).
Here we describe the process of performing commercially available ELISAs for six validated GVHD proteins: IL-2Rα5
, and REG3α3
(also known as PAP1) in a sequential fashion to minimize freeze-thaw cycles, thawed plasma time and plasma usage. For this procedure we perform the ELISAs in sequential order as determined by sample dilution factor as established in our laboratory using manufacturer ELISA kits and protocols with minor adjustments to facilitate optimal sequential ELISA performance. The resulting plasma biomarker concentrations can then be compiled and analyzed for significant findings within a patient cohort. While these biomarkers are currently for research purposes only, their incorporation into clinical care is currently being investigated in clinical trials.
This technique can be applied to perform ELISAs for multiple proteins/cytokines of interest on the same sample(s) provided the samples do not need to be mixed with other reagents. If ELISA kits do not come with pre-coated plates, 96-well half-well plates or 384-well plates can be used to further minimize use of samples/reagents.
Medicine, Issue 68, ELISA, Sequential ELISA, Cytokine, Blood plasma, biomarkers, proteomics, graft-versus-host disease, Small sample, Quantification
Analytical Techniques for Assaying Nitric Oxide Bioactivity
Institutions: University of Texas Health Science Center at Houston , Baylor College of Medicine .
Nitric oxide (NO) is a diatomic free radical that is extremely short lived in biological systems (less than 1 second in circulating blood)1
. NO may be considered one of the most important signaling molecules produced in our body, regulating essential functions including but not limited to regulation of blood pressure, immune response and neural communication. Therefore its accurate detection and quantification in biological matrices is critical to understanding the role of NO in health and disease. With such a short physiological half life of NO, alternative strategies for the detection of reaction products of NO biochemistry have been developed. The quantification of relevant NO metabolites in multiple biological compartments provides valuable information with regards to in vivo
NO production, bioavailability and metabolism. Simply sampling a single compartment such as blood or plasma may not always provide an accurate assessment of whole body NO status, particularly in tissues. The ability to compare blood with select tissues in experimental animals will help bridge the gap between basic science and clinical medicine as far as diagnostic and prognostic utility of NO biomarkers in health and disease. Therefore, extrapolation of plasma or blood NO status to specific tissues of interest is no longer a valid approach. As a result, methods continue to be developed and validated which allow the detection and quantification of NO and NO-related products/metabolites in multiple compartments of experimental animals in vivo
. The established paradigm of NO biochemistry from production by NO synthases to activation of soluble guanylyl cyclase (sGC) to eventual oxidation to nitrite (NO2-
) and nitrate (NO3-
) may only represent part of NO's effects in vivo
. The interaction of NO and NO-derived metabolites with protein thiols, secondary amines, and metals to form S-nitrosothiols (RSNOs), N-nitrosamines (RNNOs), and nitrosyl-heme respectively represent cGMP-independent effects of NO and are likely just as important physiologically as activation of sGC by NO. A true understanding of NO in physiology is derived from in vivo
experiments sampling multiple compartments simultaneously. Nitric oxide (NO) methodology is a complex and often confusing science and the focus of many debates and discussion concerning NO biochemistry. The elucidation of new mechanisms and signaling pathways involving NO hinges on our ability to specifically, selectively and sensitively detect and quantify NO and all relevant NO products and metabolites in complex biological matrices. Here, we present a method for the rapid and sensitive analysis of nitrite and nitrate by HPLC as well as detection of free NO in biological samples using in vitro
ozone based chemiluminescence with chemical derivitazation to determine molecular source of NO as well as ex vivo
with organ bath myography.
Medicine, Issue 64, Molecular Biology, Nitric oxide, nitrite, nitrate, endothelium derived relaxing factor, HPLC, chemiluminscence
One-step Metabolomics: Carbohydrates, Organic and Amino Acids Quantified in a Single Procedure
Institutions: Saint Louis University School of Medicine.
Every infant born in the US is now screened for up to 42 rare genetic disorders called "inborn errors of metabolism". The screening method is based on tandem mass spectrometry and quantifies acylcarnitines as a screen for organic acidemias and also measures amino acids. All states also perform enzymatic testing for carbohydrate disorders such as galactosemia. Because the results can be non-specific, follow-up testing of positive results is required using a more definitive method. The present report describes the "urease" method of sample preparation for inborn error screening. Crystalline urease enzyme is used to remove urea from body fluids which permits most other water-soluble metabolites to be dehydrated and derivatized for gas chromatography in a single procedure. Dehydration by evaporation in a nitrogen stream is facilitated by adding acetonitrile and methylene chloride. Then, trimethylsilylation takes place in the presence of a unique catalyst, triethylammonium trifluoroacetate. Automated injection and chromatography is followed by macro-driven custom quantification of 192 metabolites and semi-quantification of every major component using specialized libraries of mass spectra of TMS derivatized biological compounds. The analysis may be performed on the widely-used Chemstation platform using the macros and libraries available from the author. In our laboratory, over 16,000 patient samples have been analyzed using the method with a diagnostic yield of about 17%--that is, 17% of the samples results reveal findings that should be acted upon by the ordering physician. Included in these are over 180 confirmed inborn errors, of which about 38% could not have been diagnosed using previous methods.
Biochemistry, Issue 40, metabolomics, gas chromatography/mass spectrometry, GC/MS, inborn errors, vitamin deficiency, BNA analyses, carbohydrate, amino acid, organic acid, urease
Hydrogel Nanoparticle Harvesting of Plasma or Urine for Detecting Low Abundance Proteins
Institutions: George Mason University, Ceres Nanosciences.
Novel biomarker discovery plays a crucial role in providing more sensitive and specific disease detection. Unfortunately many low-abundance biomarkers that exist in biological fluids cannot be easily detected with mass spectrometry or immunoassays because they are present in very low concentration, are labile, and are often masked by high-abundance proteins such as albumin or immunoglobulin. Bait containing poly(N-isopropylacrylamide) (NIPAm) based nanoparticles are able to overcome these physiological barriers. In one step they are able to capture, concentrate and preserve biomarkers from body fluids. Low-molecular weight analytes enter the core of the nanoparticle and are captured by different organic chemical dyes, which act as high affinity protein baits. The nanoparticles are able to concentrate the proteins of interest by several orders of magnitude. This concentration factor is sufficient to increase the protein level such that the proteins are within the detection limit of current mass spectrometers, western blotting, and immunoassays. Nanoparticles can be incubated with a plethora of biological fluids and they are able to greatly enrich the concentration of low-molecular weight proteins and peptides while excluding albumin and other high-molecular weight proteins. Our data show that a 10,000 fold amplification in the concentration of a particular analyte can be achieved, enabling mass spectrometry and immunoassays to detect previously undetectable biomarkers.
Bioengineering, Issue 90, biomarker, hydrogel, low abundance, mass spectrometry, nanoparticle, plasma, protein, urine
Multielectrode Array Recordings of the Vomeronasal Epithelium
Institutions: Washington University School of Medicine.
Understanding neural circuits requires methods to record from many neurons simultaneously. For in vitro
studies, one currently available technology is planar multielectrode array (MEA) recording. Here we document the use of MEAs to study the mouse vomeronasal organ (VNO), which plays an essential role in the detection of pheromones and social cues via a diverse population of sensory neurons expressing hundreds of types of receptors. Combining MEA recording with a robotic liquid handler to deliver chemical stimuli, the sensory responses of a large and diverse population of neurons can be recorded. The preparation allows us to remove the intact neuroepithelium of the VNO from the mouse and stimulate with a battery of chemicals or potential ligands while monitoring the electrical activity of the neurons for several hours. Therefore, this technique serves as a useful method for assessing ligand activity as well as exploring the properties of receptor neurons. We present the techniques needed to prepare the vomeronasal epithelium, MEA recording, and chemical stimulation.
JoVE Neuroscience, Issue 37, electrophysiology, multielectrode array, accessory olfactory system

domain: biology


gimmeLIVE presents Aaron Neville at North Shore Music Theatre and MAGIC 106.7 wants to send you to the show! Aaron Neville's velvety croon is as instantly recognizable as any voice in modern music. From his signature 1967 hit, "Tell It Like It Is," to his chart-topping duets with Linda Ronstadt on "Don't Know Much" and "All of My Life," Aaron has enjoyed a remarkable career. He has collected 16 Grammy nominations and was twice named Best Male Singer in the Rolling Stone critics' poll. Charles Neville's Grammy winning saxophone work spans virtually every musical genre and he has played with a who's who of musical giants. Over the past five decades, the indelible spirit of New Orleans has been synonymous with the musical dynasty known as the Neville Brothers. Throughout their long solo careers, Aaron and Charles Neville have kept this spirit alive and the Quintet's vibrant shows are the stuff of legend.
Simply log in below for your chance to win a pair of tickets to the show!


text:
Current situation in the Mekong Delta
Many travellers are concerned about the current flooding situation in Southeastasia. There is a lot of water in the Mekong Delta region in Vietnam at this stage, which is quite usual for this time of the year. The people there are used to this and it is quite important for them, since the soil gets its nutrition from the floodings and there is more fish available.
The water line this year is higher than the previous years. A few roads and some houses or small villages close by the river are flooded. However, tourism activities and our tours are not affected by the water and take place as usual.

domain: travel


text:
POLICE were this week hunting an organised crime gang who are again trying to dupe motorists into handing over cash in a fake gold jewellery scam.
Officers issued the warning after two men flagged down drivers near the M74 motorway slip road at Canderside Toll on Saturday.
The crooks tell motorists that they have run out of petrol, have no money and are either trying to visit a sick family member or taking their wife, who they say is in labour in back of the car, to hospital.
To obtain money, they offer items of worthless jewellery.
The Advertiser exclusively revealed last September how the heartless gang conned two drivers into parting with £100 each in exchange for worthless jewellery on the southbound slip road at Canderside Toll, junction 8 of the M74.
This time the fraudsters appear to have fled empty-handed.
Several members of the public who were targeted on Saturday alerted police.
The suspects are described as in their 30s or 40s with dark complexions. They claim to be a variety of different nationalities and speak English with a foreign accent.
Their vehicle is described as either a black BMW or Nissan Primera.
Sergeant Claire Campbell, of Strathclyde Police, has warned motorists who are flagged down not to stop or hand over any money and instead to contact the police.
“Unfortunately, several people were duped by a similar scam last year,” she said.
“It is extremely dangerous to stop on a motorway or any of its slip roads. If you see someone who you think is in need of assistance, then please contact the police when it is safe to do so. “Although the men are currently using a car similar to a BMW or a Nissan Primera, it is possible they may change vehicles to avoid detection.”
Anyone with information on the gang should contact police on 483300 or call Crimestoppers on 0800 555111, where calls are free and callers can remain anonymous.


text:
EAST RUTHERFORD, N.J. (AP) — The New York Giants are heading back to the University at Albany for training camp for at least another year.
The Giants and the university announced Tuesday that the new deal includes a two-year option to train in New York's capital through 2012.
There has been little doubt that the Giants would return to Albany this year, but the two sides wanted to wait until an agreement was signed before announcing it.
Camp will open on Aug. 1, with the players reporting in the morning and practicing in the afternoon.
"I can't say enough about the cooperation we receive from the university and its staff," Giants coach Tom Coughlin said. "The spirit of a common purpose and objective allows our team to get the work done that is necessary for us to compete at the highest level at the outset of the season."
The Giants have trained at the university, about 150 miles north of the Meadowlands, since 1996. This will be their 15th season in Albany, their longest period at one site in the team's training camp history.
"Albany has been an ideal summer home for our football team," Giants president John Mara said.
The Giants' average attendance since moving to Albany from Fairleigh Dickinson-Madison in New Jersey has been more than 33,000 fans. The attendance last year was a record 46,960.
This camp will be shorter than most of the previous camps, with the team leaving Aug. 20. The team, which missed the playoffs with an 8-8 record last season, will practice 16 days, including seven double sessions.
"The New York Giants' training camp at UAlbany has become part of the fabric of the Capital Region," Albany Mayor Gerald D. Jennings said. "The countless number of fans that live in Upstate New York have an opportunity to watch and interact with players, and, ultimately, form a greater bond with their team."

domain: sports


text:
Good Night Soccer includes soccer fields, soccer balls, passing, kicking, dribbling, heading the ball, shooting, goalkeeping, scoring, playing defense and offense, soccer competition, soccer moms and dads, equipment and clothing, stadiums, fans, referees, and more. It's soccer time!
This book is part of the bestselling Good Night Our World series, which includes hundreds of titles exploring iconic locations and exciting themes.
This charming board book introduces young children to the popular game of soccer. Little ones will be zigzagging around the soccer field while learning about this exciting sport.

domain: library


text:
Farm to Cup Story
Guatemala’s tropical climate is ideal for growing delicious, full-bodied coffee. That’s why we’ve chosen to bring you a Guatemalan coffee that’s medium roasted to sweet perfection. Unlike it’s dark-roasted counterpart 3 Volcanoes, Snowmageddon is sweet and rounded out by a full-bodied mouth-feel. In the cup, you’ll taste the sweetness of hot cocoa paired with cashew and mellow citrus undertones—flavor notes that are sure to please your taste buds and keep you coming back for more as refuge from the bitter cold of winter.
While we experience winter from December-March, Guatemala experiences its dry summer during this same time. So, while we shiver in the cold, the beans we love to drink flourish in the warmth. And, unlike some of their heirloom relatives, many of the varieties of coffee that are commonly grown here (such as Bourbon, Catuai, and Caturra) are known for their hardiness and versatility.
Located in the scenic Sacatepéquez region, this coffee comes from various farmers located in gorgeous Antigua, Guatemala. Here, local farmers grow, harvest and sort their coffee. The farmers then visit the San Miguel mill where the coffee is processed using a washed process. This method ensures clean, crisp flavors without running too high of a risk of acquiring too much flavor from fermentation.

domain: agronomy


- Press Release
- Nov 25, 2022
To the Moon? Yes, But why not use the Space Elevator?
Tech Central pundit Glenn Harlan Reynolds muses in his piece “Is This the Right Way to Return to the Moon?” on NASA’s $104 billion announcement on Monday for a return to the Moon by 2018 and why the money would be better spent on the Space Elevator.
I’m all for the President’s Vision for Space Exploration however the implementation of it appears to be on a rocky road. have a look at this news item by Keith Cowing of SpaceRef, “NASA Revives Apollo – While Starving Space Life Science“.


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Why Relationships Matter
Membership in voluntary associations is relatively high in German-speaking countries and these provided further chances for possible partners to meet. Both individuals should, for starters, feel confident that their partner is willing to devote time and attention to the other. This book will console a person who is going through heartbreak.
About the Author
Learn to use your brain to think sometimes. College students in their sophomore to junior year who have not been in a relationship feel anxious that they are falling behind amongst their peers. They may have also impacted you in ways that you don't realize yet. Sure, opposites sometimes attract but more often than not they repel. Wealth does not define a man.
Eventually, I would agree to an arranged marriage with a man I would never love. Couples are usually wedded through either an arranged marriage or love marriage. Because of that, you are throwing away responsible suitors who ask for your hands in marriage. Going out on a limb can be roller-coaster scary because none of us want to be rejected or to have our heart broken. British writer Kira Cochrane, online after moving to the U.
If you are an educated lady, search for better jobs or businesses you can do to cater to your needs. For me, the main difference between Brazilians and Americans is that you are less dependent when it comes to family. Love marriages are those in which the individuals have chosen a partner whom they like by their own choice prior to marriage, and usually occur with the consent of parents and family. People over thirty, lacking the recency of a college experience, have better luck online finding partners. People usually socialize on online social apps, and more likely to get attached with the people they connect online.
Life is not always like that. Dating customs and habits vary considerably throughout the world. Disappointment can create discouragement affecting other areas of our lives.
Behavior patterns are generally unwritten and constantly changing. It is time to wake up to reality. Which seemed to make him keener. Breakup Separation Annulment Divorce Widowhood.
As with most things, follow your gut. It is important to discuss relationship boundaries with the person you are dating, and be clear in your communications with them. Everyone is perfect in their own ways, but trying to Impress your love you need to be the women your man love till eternity.
All you need to do is to open your heart to love whoever God has appointed for you. Humans have been compared to other species in terms of sexual behavior. When you're developing a relationship with someone, you should hear from them with some sort of consistency that feels predictable and comfortable for you and both your lifestyles. Centers for Disease Control.
25 ESSENTIAL DATING TIPS FOR HER
What if we didn't enter a relationship from a place of need, but instead from a desire for discovery? Love is one of the most profound emotions known to human beings. He must live in an apartment and must have a car, a good one. By Samantha Smithstein Psy. The meeting can be in-person or live as well as separated by time or space such as by telephone or email or chat-based.
Some of the most heartbreaking relationship problems can be caused by lack of communication. Compatibility or restraint? There are many kinds of love, but most people seek its expression in a romantic relationship with a compatible partner. Clerics run officially sanctioned internet dating agencies with strict rules. More modern approaches such as blind dates, speed dating and dating websites are not as popular as abroad, and are not considered very effective by the majority of the population.
Naturally in life everyone needs a good romantic relationship. If the texting in your relationship is done mutually and reciprocally, then it's most likely on the healthier side. Speed dating is a fast and comfortable way to meet people. If any of this is unclear to you, at any point in the dating process, you should discuss it with your partner. Although Britons are familiar with the term dating, the rituals surrounding courtship are somewhat different from those commonly found in North America.
Tips on Relationships and Dating
You will stay long o, sister. But forever I would remember that afternoon at the canal and the possibility of love. However, the majority still takes getting into a relationship seriously.
- Strong relationships are continually nurtured with care and communication.
- Meeting through friends was also commonly cited by those in the National Health and Social Life Survey, co-directed by sociologist Edward Laumann of the University of Chicago.
- Failed relationships happen for many reasons, and the failure of a relationship is often a source of great psychological anguish.
- Matchmaking is an art based entirely on hunches, since it is impossible to predict with certainty whether two people will like each other or not.
Cars extended the range of dating as well as enabled back-seat sexual exploration. There are considerable differences between social and personal values. Who teens rely on for help, and when, may predict when they reach milestones of adulthood. Most people have to work consciously to master the skills necessary to make relationships endure and flourish.
- It is the unrealistic expectations about men you are still holding.
- Feelings sometimes can make you weak and vulnerable it is important to mask your emotions so that you can protect your loved ones with a strong heart.
- Emojis are one of my pet peeves.
- The reasons for dating in Korea are various.
The purpose of the meeting is for the two persons to decide whether to go on a date in the future. You even packed the car right there and gave him the key to sort it out. He must look for ways to fix it. Remember the bad texter I mentioned before? He is engaged in a way that lets me know he's thinking of me even when we're not together, fun facts about online dating and it's allowing me to develop feelings for him.
However, if you are dating a person and neither of you have agreed to date exclusively, then you are not in a relationship and you are both free to also date other people. Relationships between students preparing for college are frowned upon by many parents in China. And actually, it doesn't have so much to do with the relationship with another person as it does with your relationship to yourself.
If you are rejected or ignored, remember that it is not about you. One way of thinking about loners is to ask whether people realize they are better off on their own, and if so, whether they are prepared to live authentically. Partners in strong relationships also feel grateful for one another, openly provide and receive affection, and engage in honest discussions about sex. For many, romantic relationships comprise one of the most meaningful aspects of life, providing a source of deep fulfillment.

domain: psychology


text:
Samsung has been preparing a replacement for its Gear smartwatch for a few months, and this time, the connected watch should adopt a rounded design.
Samsung's next Gear smartwatch will be round. It is a press release retrieved in advance by Android Central which shares the information.
Announcing the upcoming release of the SDK of the next Samsung Gear watch, the press release is accompanied by visuals offering a display in the form of discs.
It must be said that despite an anticipated foray into the smartwatches market, Samsung found that its main competitors opting for a round display sold more easily than its own watches.
And the trend quickly spread with the Android Wear platform, with smart watches more widely opting for a traditional style.
With the release of the Apple Watch, Samsung would also need to stand out from Apple by offering a watch with a clearly different design in order to avoid comparisons.


96kHz/24-bit, FLAC; Decca 4788758 (supplied by www.highresaudio.com)
Glazunov’s romantic concerto provides a balm after the terrors and intensity of the Shostakovich: held back from performance until Stalin’s dictatorship was over. There was far slighter music by that composer – ‘Romance’ from The Gadfly – in Benedetti’s 2012 Decca collaboration with the Bournemouth Orchestra and its Ukranian chief conductor, The Silver Violin. Here, she rises to the challenges of music written for Oistrakh, whose 1955 New York benchmark recording is now on Sony. There’s something of a Heifetzian concentration in her playing (and the coupling sounds even more like a live performance) so it’s a shame she’s given full stage width for the long cadenza and we briefly get a ‘wrong end of a telescope’ effect as the Burlesque opens, although the ear quickly adapts. CB
Sound Quality: 90%
Hi-Fi News Lab Report
The anomalous tone at 20kHz is more irritating from a technical than audible standpoint [see Graph], otherwise this 96kHz rendering is free of compression and makes good use of its bandwidth to capture the thrilling string harmonics. PM
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text:
My husband (Rob) and I (Kerry) are 50+ professionals with a daughter 19 (Sarinah) and a son 16 (Jesse). We live in sunny Western Australia within walking distance of the beautiful Indian Ocean. We love travelling and meeting people from different cultures. We speak English, Indonesian and a little German.
About Our Home
Our large air conditioned family home has four double bedrooms and two spacious bathrooms. There is plenty of room for everyone whether relaxing in the large lounge, the comfortable family room or sunning by the sparkling pool. Those who like to cook will find the large kitchen meets all their needs with a double oven, gas hotplates, microwave and dishwasher. Outdoors there is a large gabled patio with comfortable furniture and,of course, the stunning salt water pool.
- Swimming pool
- Outdoor entertaining area
- Off-street parking
- Outdoor garden
The white sandy beaches of the Indian Ocean are within walking distance and you may participate in many water sports including kitesurfing, windsurfing, boating, fishing and snorkelling. There is easy access to public transport and Perth city is only 20 mins by train.
- Movies/ Theatres
- Public transport
- Parks/ Gardens
Within walking distance of shopping centres and restaurants.
Not yet a member? Register Today and receive the following great benefits
- Choose from thousands of homes located in premier holiday destinations around the world!
- Swap your house and easily save hundreds or thousands of dollars on holiday accommodation.
- Receive the best travel tips from locals who really know the area because they live there!
SAY GOODBYE TO HOTEL BILLS FOREVER WITH HOUSE SWAP HOLIDAYS!
Join Australia's home exchange community today - membership is only $59.95 for 12 months.Get started today and save!
Preferred Travel DestinationsGermany, France, Austria, Switzerland, Italy, Spain
Preferred DatesApril, May, June, July
Closest AirportPerth International (40 kms)
Languages SpokenEnglish, German
Other DetailsNo smoking
Melbourne, Victoria, Australia
Open plan with large living areas at one of Port Phillip Bay's best beaches. Has hosted many family holidays and events. read more
Ettalong beach , New South Wales, Australia
Delightful 1 bedroom apartment within the mantra complex at Ettalong beach read more
Christchurch, Canterbury, New Zealand
Comfortable, country living in lovely area with space to enjoy relaxing read more

domain: geography


text:
Avengers and Co. Create Vines - first chapter
166 days ago
I will write Avengers (and other characters) one-shots. They will usually consist of Avengers, vines, memes, and fan art. I get most fan art from google images, but I´ll try to give credit to the artists if I can. You can request vine stories or link me fan art to write a story about. Just comment your request. I still make my own ideas.

domain: artistic


text:
Henriette Fireplace February 17th, 2018 - 07:41:25
Even ventless gas fireplace fire boxes come with a spark arrestor curtain, chain-mail screen to add to the illusion of a wood burning fireplace with the convenience of a gas log installation and remote control use. Custom fireplaces will have inconsistencies in width, height and fireplace depth. While fireplace screens are produced in standardized sizes, not all fireplaces will be that exact and the fire-screen may over-shoot the size. Spark arrestor curtains can be custom made for specific rods to fit inside any custom fireplace opening but free standing fireplace screens are available in standard finished and colors. When selecting a fireplace screen most often the room décor, and style and fitting the size of the fireplace are primary considerations. A fireplace screen can be as ornate or as simple as the style of the room dictates. A fireplace screen can add to the beauty of a room while assuring that the unprotected floor or carpeting is safe from flying sparks and the fire itself kept apart from curious pets and small children.
5. Put Your Fireplace Candelabra on Your Mantel - For those of us who have gas or electric fireplaces, and do not wish to remove the fake logs during the warm season, consider placing a candelabrum or two on your mantel. This is actually a very classic way of decorating your fireplace. Use fireplace candelabra that are made for smaller fireplaces or zero clearance fireplaces. 6. Put Your Fireplace Candelabra on Your Hearth - If you do not have a mantel, you can place your fireplace candelabra on your hearth just as easily. Be sure to add a little greenery in the spring, maybe a sting of fall leaves in the autumn and some pine needles and cones in the winter for an added special touch.
Fireplace screens can serve as spark arrestors for the wood burning fireplace and as decorative items or they can help protect a hearth floor area from rolling logs moving around as flames burn off a wood stack in the case of sturdier types of screens. Most screens are made of metal such as brass or more commonly wrought iron or steel. Since fireplace screens are subjected to the full heat of the fireplace they must be made of durable materials. Perhaps the most common type of fireplace screen is the mesh spark arrestor made of finely wrought interlinked chains, which are hung from a rod suspended from the lintel inside the fireplace opening. The screens can be drawn shut like a curtain when needed or opened when the fireplace is not in use.
4. Take Your Fireplace Candelabra Outside - Candelabra made for your fireplace are usually pretty sturdy. They are generally made out of wrought-iron or some other very durable material. This makes them ideal for use outside. Try using your fireplace candelabra as a centerpiece on your patio table. If you have a long picnic table, you can use a candelabrum that is designed for a smaller fireplace. How about using a couple candelabra to decorate either side of your front door, or back door? If you are worried about your candles blowing out from gusts of wind, you can use votive candles surrounded by glass, but your candelabra cannot have center pins for this to work.


text:
400 m ühistranspordini
|Lähim lennujaam||Hongkongi rahvusvaheline lennujaam (HKG) - Hongkong|
|Kaugus lennujaamast||29,2 km|
Newton Place Hotel, asudes paigas Kowloon ida, Hongkong, on reisijatele populaarne valik. Siit võivad külalised nautida lihtsat ligipääsu kõigele, mida pulbitseval linnal pakkuda on. See modernne hotell on linna popimate vaatamisväärsuste, nagu GOME Mall (Kwun Tong Road), é¦™æ¸¯Apm, Kwun Tong, läheduses.
Newton Place Hotel's muudavad laitmatu teenindus ja ülihead võimalused Sinu siinviibimise unustamatuks. Kui nimetada vaid mõni hotelli võimalustest, oleksid need 24-tunnine turvamine, igapäevane toateenindus, ratastooliga ligipääsetav, 24-tunnine vastuvõtt, vahendid erivajadustega külalistele.
Kõigi majutuste tunnusjooneks on läbimõeldud valik mugavusi, et tagada külalistele võrratu heaolutunne. Pealegi, hotelli vaba aja veetmise info tagab, et Sul siinviibimise ajal ettevõtmistest puudu ei tuleks. Avasta kütkestav segu Newton Place Hotel professionaalsest teenindusest ja mitmekesistest teenustest.
Lapsed and lisavoodid
Lisavoodi võimalus sõltub valitud toast, lisainformatsiooni saamiseks vaata konkreetse toa mahutavust.
Külalised peavad olema vähemalt 0-aastased.
Väikelapsed vanuses alates 0 kuni 1 aastat [kaasa arvatud]
Viibimine tasuta, kui kasutatakse olemasolevat magamisvarustust.
Beebivoodi/-häll soovi korral saadaval
Lapsed vanuses alates 2 kuni 12 aastat [kaasa arvatud]
Viibimine tasuta, kui kasutatakse olemasolevat magamisvarustust.
Kui vajad lisavoodit, küsitakse selle eest lisatasu.
13-aastaseid ja vanemaid peetakse täiskasvanuks.
Peavad kasutama lisavoodit, mille eest tuleb maksta lisatasu.
Newton Place Hotel
Kuva arvustusi, mis mainivad järgmist
accessible and convinient”
I had a pleasant stay at the hotel. The location to very convenient and accessible to all means of transportation. I will definitely come back to this hotel. The staffs were very helpful and attentive to my queries.
GOOD BUSINESS HOTEL”
The check-in was speedily efficient by personable, well-dressed staff. The bed, and shower were excellent, and the room clean with all the required amenities. The check-out was promptly done, also by pleasant staff. I will stay in this hotel again.
lovely hotel ”
room service is very good and clean very tidy . the hotel lobby finishing is attractive, and have his own style. the service counter also very friendly ,and effiency. whatever you want to know about the hotel facility ,or about the nearby shopping area ,she will detail introduce to you'.actually nearby is a lot good shopping shops.
Business trip to H.K.”
Stayed here the first time on a business trip and was over all very please with the hotel. Staff trying to be very helpful. Clean room, they even recognized that international people prefer a different kind of breakfast room than local Chinese people. only drawback would be the location of the hotel and that you need to order the free shuttle bus to the express train station at least four hour in advanced.
First Visit at a hotel at Kung Tong”
The hotel is located in an industrial area but the inside of the hotel is very nice. The services staff is very friendly and helpful. The breakfast buffet is very nice andits price is very reasonable. I consider staying this hotel again when I visit Hong Kong again.
value for money”
well! i have stayed once and this time round i choose the hotel again because is near my relative house. though is far from town but is walking distance from the mrt about 10 minutes. the room is nice and new. i feel comfortable of course. the hotel staff is friendly and they can speak fluent english. the room rate is not expensive and i will stay in the hotel again.
we enjoy our stay in this hotel though is surrounded by industrial building because the price is reasonable and 7minutes walking distance to the mtr. a few food stall is nearby. the most important is that the hotel's room is very clean and comfortable because is only 1 year old. the hotel staff is also very friendly and helpfull.
Its was a simple n nice hotel, minutes away from Kai tak Cruise Terminal. We were given a room with full view of Genting Dream Cruise that we disembarked from the terminal. The view was awesome.!! Location to MTR, shopping mall and eateries were convenient. Will stay in the near future when we are back in Hong Kong.
Well.. think for the amount spending, this consider quite a decent hotel. Staff is friendly but having trouble checking in due to wrong name given by agoda, time solving is very very slow, took few hour. Don't intend to say furthermore.
Value for money”
Value for money, room space is bigger than I thought, deco is ok but not to 4 star standard. Location is ok, 10 mins walk to the subway station. We got lost by just asking people along the street after getting off the station, it seems that less people knew about this Hotel, suggest review the map before you go.
room had an extremely strong smoking/cigarette odour - so bad it was burning my eyes. couldn't change room because the hotel was full. bed was not very comfortable, it was like one you would find in a cheap one night hotel. this is probably what this hotel is, as most of the people checking in looked like they were on tour groups. this is not a good hotel if you plan to stay for more than 1 night and will spend a lot of time in the room.
I was annoyed and truly disturbed by the China Mainlanders. I think this hotel is only designed for the mainland china people. They are noisy on the corridor and the room is not well sound proofed. The rooms are small, beds are small.
nice and enjoy”
every time when i return to this hotel, i feel like going into the new room again. the service is supered. It is very near to my client office therefore, by walking it takes only 15mins. will sure stay in this hotel again if coming back to HK.
The price was decent for the location”
The room conditions were alright but the view was out of my expectation because I would least expect to have such a view from the location away from the city and shopping district. I chose this hotel for its price and therefore I would recommend anyone who knows how to get around in Hong Kong for a stay because it saved us alot of money compared with living near the shopping/tourist area.
Poor Price, Poor Location and Poor Service”
If you use this hotel for business, then I guess is ok consider it is close to a lot of industrial buildings. The staff has no smile, other guests are quite noisy and especailly in the morning. Very bad bathroom and interior. Do not recommend.
Surprising Harbour View”
it was surprised when i stepped in my room when i saw the whole harbour view in front of me. the room is clean and bright. room is reasonable big. the bathroom with glass window. it's located around the commercial/industrial area but short walk to mtr station. there's new trendy shopping mall (apm) and also plenty local eating places. many buses you can jump into and get to anywhere in hong kong. i'm very happy of the stay. if they can provide free in-room wi-fi will be perfect.
Hotel location is far away from downtown”
the hotel is ok to stay if you need to stay in kwun tong to work. service is ok and not too far away from mtr. it's pretty far away from downtown area and nowhere to go at night. it's a business area. but pricewise compare with other, it's acceptable.
not a bad hotel”
this is actually not a bad hotel for business travellers who have business appointments at this commercial cum industrial districts of kwun tong, kowloon. i guess this must be the best hotel in kwun tong. it provides free internet service at the hotel lobby, and one can even get personal help from the hotel staff to log-on to internet, or to check-in air tickets on-line. it's only a few minutes walk from the hotel to a big shopping mall with jusco supermarket on its lower ground floor. there are many different choices of good local & western restaurants and food courts in the mall too. i'll stay at this hotel again if i come to kwun tong the next time.
Good Hotel but a bit out of the centre.”
There is one shopping mall local to the hotel and some good local restaurants. It is about a 10 minuit ride to the centre but this was not a problem. If you don't mind usng the train then it is very easy.
Room Tidy & Clean. Even small but comfortable. Ve”
i have good impression. very silent environment and no worry for the food & drink 24hrs as many different restaurants around the hotel even the hotel have no restaurant facility itself. it is situated near mtr station that traffic is convenient. room is small but very tidy & clean that it is comfortable. and the staffs are so helpful & have smiling faces. they are very polite.