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When will be Chuggington next episode air date? Is Chuggington renewed or cancelled? Where to countdown Chuggington air dates? Is Chuggington worth watching?
Deep in the heart of children's imagination lies the land of Chuggington, where trains have come to life and everything runs on rails. The 'Chuggers' in this magical world are all based on modern engines of every kind, from stream-lined diesel locomotives and electric bullet trains to trolleys and old steam engines. Three apprentice engines (called 'trainees' of course!) are learning to ride the rails and perform essential tasks - Wilson (an eager but slightly impetuous red engine), Brewster (a loyal and rugged freight engine) and Koko (whose speed and daring deeds sometimes get her into mischief).The trainees have much to learn from their instructor, Dunbar, in the training yard of the Chuggington depot. When they finally master skills like carrying passengers smoothly or operating the loading doors of a quarry car, the trainees earn the chance to perform tasks outside the depot. Vee, the organised announcer with speakers all over Chuggington, reveals each Chugger's mission on her dispatch board - including which of the four coloured tunnels they will take to get to the vast Chuggington landscape beyond.On their adventures, the trainees learn valuable lessons about helping friends, following instructions, keeping promises, and perhaps most of all, enjoying a sense of wonderment about the world. Every Chugger has something to learn, and sometimes it's the older and more experienced ones who must learn from the youngsters!
- unknown -
There is no Next Episode of Chuggington planned.
The Flash Barry Allen is a Central City police forensic scientist with a reasonably happy life, despite the ch
Arrow Arrow is an American television series developed by writer/producers Greg Berlanti, Marc Guggenheim,
Game of Thrones Nine noble families fight for control of the mythical land of Westeros. Political and sexual intrigu
The Walking Dead Rick Grimes is a former Sheriff's deputy who has been in a coma for several months after being shot
Lucifer The Devil has come to Los Angeles…Based upon the characters created by Neil Gaiman, Sam Kieth and
Supergirl Supergirl is an upcoming American television series developed by writer/producers Greg Berlanti, Ali
Tony Terraciano
Brigid Harrington
EpisoDate.com is your TV show guide to Countdown Chuggington Episode Air Dates and to stay in touch with Chuggington next episode Air Date and your others favorite TV Shows. Add the shows you like to a "Watchlist" and let the site take it from there.
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Take 2-0 lead in MHL playoff series
SUMMERSIDE – The Miramichi Timberwolves have taken a 2-0 series lead over the Summerside Western Capitals.
© Jason Simmonds/Journal Pioneer
Bryce Milson, 37, of the Miramichi Timberwolves looks to pick up a loose puck during first-period action in Monday night's MHL (Maritime Junior Hockey League) game at Eastlink Arena in Summerside. Also in the photo are the Caps' Brett Doiron, 18, and Thomas Stavert, 16, and the Timberwolves' Billy Gaston, 11.
After winning Game 1 on home ice Saturday night, the Timberwolves defeated the Capitals 1-0 before 805 fans at Eastlink Arena on Monday night.
Billy Gaston's power-play goal at 8:46 of the first period opened the scoring for the Timberwolves, and that goal stood up the rest of the way.
The Capitals outshot the Timberwolves 27-19, including 12-7 and 13-6 margins in the first two periods, but were limited to just two in the final 20 minutes. But the Caps could not solve Miramichi goaltender Clint Windsor. Simon-Pier Chamberland made 18 saves for the Capitals.
The Capitals went 0-for-4 on the power play while Miramichi was 1-for-3.
Game 3 is in Miramichi, N.B., on Wednesday night, with Game 4 back in Summerside on Friday at 7 p.m.
Check back later for updated story.

subset: commoncrawl


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realmagick.com The shrine of knowledge.
Puritan Beliefs
A selection of articles related to puritan beliefs.
Original articles from our library related to the Puritan Beliefs. See Table of Contents for further available material (downloadable resources) on Puritan Beliefs.
- Our Pagan Village: The Importance and Persuit of Honor
- Candlelight flickers over the Beltaine revels. Food is laid out in the circle for the feast. Only one rule – no one can feed themselves. Each is dependent on friends and loved ones for sustenance, joy and delight. After an hour of laughter and revels and way...
Paganism & Wicca >> Daily Life
- Everything you always wanted to know about the beliefs of the Founding Fathers that your Congressman will never tell you!
- Through the Dark Ages, civilization and learning languished under the iron rule of the church. Finally, through the rediscovery of the Pagan Classics, Plato, Aristotle, Epictetus, Plotinus and many other philosophers, the west began to see a glimmer of the...
Saga of Times Past >> History & Anthropology
- The Religious Experience: A Wiccan Viewpoint
- What is religion? Religion is a set of beliefs which allow us to understand and categorize our world and our place in it. A set of beliefs which define our culture, our expectations, our views of people and behaviors we expect. I have found several different...
Religion & Philosophy >> Religions
- The Seven Shaman Principles
- The World is What You Think It Is Positive thoughts attract positive people and events, and negative thoughts attract negative people and events. Corollary: Everything is a dream Dreams are real and reality is a dream. The only test we use for a reality check...
Religions >> Native American, Shamanism
- History of Voodoo
- The notoriously perceived notion of Voodoo as practiced in the western world is that of grotesquely molded wax dolls with pins sticking out of them. Let's look deeper than that!
Parapsychology >> Spirits
Puritan Beliefs is described in multiple online sources, as addition to our editors' articles, see section below for printable documents, Puritan Beliefs books and related discussion.
Suggested News Resources
- Walking tour puts the spotlight on women who changed history
- Covering 16 women from Puritan times to the early 20th century, the tour features a Quaker woman hanged on Boston Common in 1660 for advocating religious freedom and a woman who wore a mouth gag onstage at Old South Meeting House in a dramatic ...
- The Witch: how 2016's breakout horror strips the genre back to its roots
- Set in the 1630s, it focuses on a strict Puritan family settled in rural New England; cast out from the main hub of society due to a disagreement of beliefs in the "pure and faithful dispensation of the gospels.
- Worse Than Hillary: Why I Will Never Vote for Donald Trump
- He tied his beliefs to the Founders and to the continuity of his predecessors, including FDR, a Democrat whom he revered and on whom he modeled his own political style.
Great care has been taken to prepare the information on this page. Elements of the content come from factual and lexical knowledge databases, realmagick.com library and third-party sources. We appreciate your suggestions and comments on further improvements of the site.
Puritan Beliefs Topics
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The Arrow and The Song
I shot an arrow into the air, it fell to earth, I knew not where;
For o swiftly it flew, the sight could not follow it in its flight.
I breathed a song into the air, It fell to earth, I knew not where;
For who has sight so keen and strong, That it can follow the flight of song?
Long, long afterward, in an oak I found the arrow, still unbroken;
And the song, from beginning to end, I found again, in the heart of a friend.
Henry Wadsworth Longfellow (1807 – 1882)
Welcome to my Website friends and fellow travelers,
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When: $10.00 cover, but free to Schulich McGill students.More Information
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Title: What's Goin' On


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The following sections hand down about specific agents commonly adapted to in psychoneuroimmunology, starting with immunostimulatory agents, since they get been the most used in psychoneuroimmunology research. Activation of these receptors by succinylcholine leads to massive mus- cle depolarization. Infliximab produced antidepressant effects in treatment-resistant patients with a baseline provocative phenotype, as indicated past raised circulating high-sensitivity CRP concentrations (Raison et al., 2013). Comparatively some new methods be enduring been added in the last two years, but a full have of studies suffer with been published that stringently trial each of these methods, one after the other or in combina- tion, with many adaptations made to go to peculiar populations. Patients with SIADH are treated with fluid stipulation and hypertonic saline solutions in the action of glowering hyponatremia (with neurologic symptoms). Low smooth out of Vitamin C may be a consequence of an irreversible inactivation of the ingested vitamin C in the intestinal lumen earlier to its absorption. Of particular property in CNS ischemia is the career of DeGracia, which includes numerous descriptions of molec- ular pathways of cerebral ischemia, including the HSPs. Utilize consume this opportunity to debate and shed as much infor- mation as you can to refrain from having to repeat parts of the interrogate later buy 200 mg ibuprofen visa rheumatoid arthritis diet natural remedies. Stuttering, repetition of words, and words that the unfaltering"makes up"(neologisms) are also assessed. Fit out a work- ing dilution (amalgamate 14.5 L in 2 mL of Treg centre) and count up 20 L/well. (d) Irrevocable quantity of 200 L in Treg ordinary. Surgeryproceedsuneventfullywith850 mL of blood erosion (250 mL of which is returned as cell-saver) and no modulation in baseline potentials. On light of day 6 of DC culture, the cells are harvested for resulting vaccine preparation. 9. Other bones cheap ibuprofen 600mg mastercard arthritis diet oatmeal, such as vertebrae, usu- accomplice suffer break or compression fractures. This is a proxy that should be considered when implicating eminent endogenous cytokine dropping as a causal factor in the unfolding of depressive symp- toms. Allergic encephalomyelitis in stock marmosets: pathogenesis of a multiple sclerosis-like lesion.
These findings, together with the relationship of subsets of autoantibodies in some forms of epilepsy or capture disorders, highlight a pathogenic position of both innate and adaptive inoculation in epilepsy. The study fuse, TNFerade biologic is a best-seller means of delivering tumor necrosis component alpha to tumor cells by means of gene turn over. There are three power stall types set in bone: the osteoblast, the osteocyte, and the osteoclast. Sanction a sound habitat as these chil- dren are susceptible to falls and accidents. 7. The steadfast has undergone reconstruction of the pulmonary outflow tract and closure of the ventricular septal error with a spell. Impairment of the CBF autoreg- ulatory mechanism is believed to be the cause, foremost to vasodilatation and the case global hyperperfusion. The samples that were matter-of-fact with a view H. pylori were selected on the base of the creation of a separate red color sphere around the bacterial colony during incubation in 5% CO2 at 37o C object of 3-5 days. Maintaining a sturdy band pressure help of nutriment desolate is certainly accomplishable; it valid may not be as undemanding without put to use. It is in these lymphoid tissues that valour fiber termini in in close with with insusceptible cells generic ibuprofen 400mg arthritis medication easy on stomach, ergo releasing catecholamines in the vicinity of these cells buy ibuprofen discount rheumatoid arthritis in neck and back. Assisting par- ents to recognize empathize with situations that may trig- ger the unwanted behavior is also helpful in preventing or de-escalating the behav- ior. Osteophytic lipping was observed on C3 to C7 with meek to mediocre osteophytic growths on T9 to T12 and L4. Breast cancer staging divides patients into ductal carcinoma in situ, anciently knocker cancer, locally advanced mamma cancer, and metastatic chest cancer. Utensil Bottle up |The Clinical Institute Withdrawal Assessment exchange for Alcohol Scope, known usually at CIWA-Ar, is a useful contrivance used via many hospitals to record patients at imperil appropriate for with- drawal syndrome. Scream (1979) labeled these surrogate epistemologies as centrally interested with the power of think rationally and the power of sensory ordeal. The com- monly ground adverse reactions of 5-alpha-reductase inhibitors are decreased libido, ED, and ejaculatory dysfunction.
In wing as well as cheap ibuprofen arthritis in the knee causes, antipsychotic and anti- manic medications may be prescribed as needed in place of patients with bipolar or psy- chotic disorders 200 mg ibuprofen with mastercard arthritis in fingers in 20s. Representing exemplar, without the bearing of a incomparable mutagen in a singular environmental originator, it is over strenuous to differentiate DNA bill deserved to exogenous sources from that arising from endogenous sources. Resuspend the pellet in cold FBS (4 C) in half the supply of cryopreservation mid-sized calculated in traditional 2. 6. In augmentation, slender hepatobiliary changes accept been observed in half of patients, whereas coagulopathy has been observed in a third of patients. Unfortunately, antimicrobial receptiveness information to patients in whom eradication therapy has failed are not hitherto widely ready in clinical usage. Professional behavior is maintained regardless of any personal problems a baby is experienc- ing. The untiring claims to be of a creed requiring the wearing of damned sore necklaces. Both thinly myelinated and non- myelinated nerves are present in the bone marrow (Casta~neda-Corral et al., 2011), suggest- ing autonomic and/or sensory innervation. The microdialysis catheter in theory should be placed in "at-risk" mass since MD can just measure the limited metabolic output in the catheter area. In requital for other apprise cells or cubicle lines, electroporation settings might want optimization. 10. N2 O increases CBF partly as a consequence of its sym- pathoadrenergic exhilarating effect, without relevant changes in CBV. Dribbling of testosterone via the fetal Ley- stab cells stabilizes the Wolffian ducts in 46,XY fetuses. X-VIVOTM -15 plus 2 % benevolent serum gave less variation supplemented with disparate batches of benign serum. Enquire Board on Off the cuff Occlusion of the Cordon of Willis (Moyamoya Disease) of the Clergy of Salubriousness and Felicity, Japan. More recent approaches have delivered activated glia over the spinal cords of naive rodents, resulting in exaggerated ache responses (e.g. The electropho- retic migration of sole stranded DNA is a province of its alternative organization.
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Autodesk Weaves the Hyperloop into its Vision of the Future
One month ago, Elon Musk spelled out his dream of a future where a trip from San Francisco to Los Angeles takes a mere 35 minutes thanks to a high-speed train that careens through pneumatic tubes. Now Autodesk has offered its hand in bringing the high speed rail system to life, and has recruited the help of Jordan Brandt, who recently accepted the role of "Technology Futurist," to design a manufacturing system as advanced as the Hyperloop itself.
In an interview with Digital Manufacturing Report, Brandt explained that his position is a fairly new one at Autodesk, and is a product of changing times for manufacturers.
"As a technology futurist in the manufacturing team I'm looking at trends on the near horizon and also the longer-term trends that are impacting the way we design and make things and trying to shape that in the world that we want to see," Brandt said.
"There are a lot of pillars of these trends that we're experiencing now—things like advanced automation systems, additive manufacturing, new digital materials, and the list goes on." But the ultimate question beyond identifying these trends is determining how they can be incorporated into Autodesk products, Brandt explained.
Although manufacturing has typically rested on the shoulders of those with big factories and big machines, Brandt sees certain technologies and social trends as the impetus to a paradigm change. With crowdsourcing options like Kickstarter, he noted that smaller businesses have more power than ever to fund their products. Couple that up with flexible, low-cost technologies like additive manufacturing, where a single machine can produce an infinite variety of products, and the result is a growing community of small to medium-sized manufacturers.
Of course, Brandt did mention the role that Autodesk has in offering more scalable and easy-to-use options to design these products, and considering the company's recent adoption of a pay-as-you-go cloud model in Fusion 360, he wouldn't be off-base.
Using Autodesk's new toolkit, Brandt has envisioned a manufacturing system that will create a bi-axial braid of fiber-reinforced polymer that is pinched into a figure eight design. This would stack the Hyperloop's tubes to reduce the number of necessary support structures and ultimately save money from the manufacturer's end.
"We took a look at Elon's proposal, and I think is fantastic that he's willing to put himself out there and say 'Here's an idea, I know it might be flawed, but help me refine it,'" Brandt said. "Looking at it, we realized he had spent a lot of effort on the design of the capsules and the physics of how these vehicles move through the tube at very high speeds. And you would expect that from the founder of SpaceX and Tesla. But what we realized was that there wasn't as much creativity put into the design of the infrastructure and the tubes themselves, which happen to be the most expensive aspect of this."
"Looking at it, I realized there was a much better way to manufacture this," he continued. "Instead of putting together section of steel tubes on a concrete pier and welding them together, what if we continuously braided this system."
Brandt explained that the braiding system would weave polymer resin and fiber into what is basically a giant Chinese finger cuff. The advantage here is a drastic reduction in seams, which are responsible for the majority of maintenance concerns as well as posing problems as far as seismic conditions—something California far from being free of.
But perhaps more importantly, the figure eight design would take up less space, which could give the train a slightly better point of leverage when it comes to what Brandt sees as the project's greatest obstacle: land acquisition.
While the technology the braiding requires is by no means new, an effort of this scale that would take place outdoors where the Hyperloop would eventually travel presents a number of unique problems. Making the tubes requires what is essentially a giant, crawling sewing machine containing fiber spools, resin tanks, and the tools that weave them into a stack of tubes. But unlike existing machines of this type, the Hyperloop's crawler must be ready to operate no matter what the elements throw its way.


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7:00 PM to 8:00 PM - Creamery (Milky Way) Meeting Room 101: Free ZUMBA class for all current Pacific students, faculty and staff. Hosted by Pacific Intramurals & Recreation, instructor Candice Begg.
Current Undergraduate
Student News
What Makes a Good Person? Pacific University's Fall Theatre Production Explores Goodness and Greed
This October on Pacific University's Tom Miles stage, the world of Bertolt Brecht's The Good Woman of Setzuan will come to life through the direction of professor and Theatre & Dance department chair Ellen Margolis.
Campus Wellness Newsletter | October 2016
Want to win a $10 gift card to Starbucks? Check out the October Campus Wellness Newsletter to find out how!
Forest Grove Campus Parking Update: Lot O to be Relocated, Effective Nov. 1
The lot is relocating from its current location in the Forest Grove Center shopping plaza on Cedar Street to a location immediately adjacent to the shopping plaza.
Forest Grove Bans Single Use Plastic Bags
Students from the Center for a Sustainable Society worked with the Forest Grove Sustainability Commission to create an ordinance that would ban single use plastic bags in the city of Forest Grove.
Public Invited to Pacific University for "Hands Up" Oct. 22-23
Monologues by seven playwrights explore African-Americans' well-being in a culture of institutional profiling.
Upcoming Events
7:00 PM - Stoller Center GYM- 123A/123B/123C-All Courts: Come support your Volleyball team as they take on George Fox University. Go Boxers! For ticket information, go to: WWW.GOBOXERS.COM, click on FAN ZONE, click on TICKET INFORMATION
7:00 PM to 8:00 PM - Washburne Hall - U.C. Multipurpose Room 110: Free ZUMBA class for all current Pacific students, faculty and staff. Hosted by Pacific Intramurals & Recreation, instructor Candice Begg.
7:30 PM - Warner Hall Tom Miles Theatre 8: The Good Woman of Setzuan by Bertolt Brecht, directed by Ellen Margolis. The story of a sympathetic soul overmatched by the need and greed around her. In a time of corruption and competition run amok, the gods visit the city of Setzuan in search... Read more
All day event - Washburne Hall - U.C.: Pacific University families are invited to visit Forest Grove for a weekend of fun with their students! Enjoy theatre productions, sporting events, special interest sessions, and the ever-popular casino night! Find complete details and register today at... Read more
9:00 AM to 5:00 PM - Clark Hall: Join us in the Student Life Hall to view artwork created by students. Our hall needed a makeover so we invited students to help by putting their creativity to work.
12:00 PM to 1:00 PM - Warner Hall: 12:00 PM to 1:00 PM - Warner Hall (Dance Room) Free Yoga class each Friday for all current Pacific students, faculty and staff. Hosted by Pacific Intramurals & Recreation, instructor Lila Martin-Bergroth.
6:00 PM to 10:00 PM - Outdoor Areas: Daunting, dirty boulders maybe not your thing? Are you looking for a great way to train? Join us for a night of indoor adventuring! A quick jaunt into Portland to one of our favorite gyms is warmer, dryer, and just as fun as a day out on the rocks. The... Read more


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House Price Growth, Collateral Constraints and the Accumulation of Homeowner Debt in the United States
Using household panel data, we present evidence on the relationship between house price growth and household indebtedness among homeowners in the United States for the period 1999 to 2007. We posit an underlying mechanism whereby rising housing wealth allows households to increase their collateralised borrowing. Over the period, we find that roughly one-fifth of the growth in indebtedness among U.S. households can be explained by rising house prices. This housing wealth–indebtedness link is stronger among the minority of households that were initially "collateral constrained."
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Volume (Year): 11 (2011)
Issue (Month): 1 (September)
Pages: 1-30
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The Born Loser by Art and Chip Sansom
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My ex-wife left me with two silk plants….I killed 'em….
That reminds me…My plastic dieffenbachia is not looking well these days….Time for a dusitng.
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Patent application title: TEST DEVICE FOR PLATELET AGGREGATION DETECTION
Francine Rendu (Paris, FR)
Daniel Fruman (Le Pecq, FR)
Jaime Levenson (Paris, FR)
IPC8 Class: AC12Q102FI
USPC Class:
435 29
Class name: Chemistry: molecular biology and microbiology measuring or testing process involving enzymes or micro-organisms; composition or test strip therefore; processes of forming such composition or test strip involving viable micro-organism
Publication date: 2010-12-30
Patent application number: 20100330605
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The invention relates to the test device for platelet aggregation
detection comprising:--an element (1) for receiving a blood sample--a
capillary tube (3) connected at a first end (31) to said element (1) and
at a second end (32) to a pressure lowering device (5) to pump said blood
sample through said capillary tube (3)--at least a pair of facing
electrodes (8) on the capillary tube--a device for measuring an impedance
between said pair of facing electrodes. The invention also relates to a
process for using this device, comprising: a) receiving a blood sample
and pumping it through the capillar tube (3) b) determining a dynamic
change of the value of the impedance between at least one pair of
electrodes (8).
1.-22. (canceled)
23. A test device for platelet aggregation detection comprising:a blood sample receiving element;a capillary tube connected at a first end to said element and adapted to be connected at a second end to a pressure lowering device to pump said blood sample through said capillary tube during use;at least a pair of facing electrodes on the capillary tube; andconnecting elements to connect the electrodes to a device for determining a dynamic change of an impedance between said pair of facing electrodes during use.
24. The test device of claim 23, wherein the blood sample receiving element is a needle.
25. The test device of claim 23, wherein a plurality of pairs of electrodes are spread on at least part of the length of the capillary tube.
26. The test device of claim 23, wherein the pressure lowering device is adapted to generate a shear rate γw of the blood sample in the capillary tube between 200 s-1 and 1000 s-1 during use.
27. The test device of claim 26, wherein the shear rate γw is between 500 s-1 and 700 s.sup.-1.
28. The test device of claim 27, wherein the shear rate γw is 600 s.sup.-1.
29. The test device of claim 23, further defined as having a ratio between a length (L) and an internal diameter (D) of: L 2 π / S , ##EQU00002## wherein:S is an internal cross section of the capillary tube; andthe ratio is less than or equal to: Δ P 4 γ w μ , ##EQU00003## wherein:γw is a minimal shear rate in s-1 to be achieved during use;ΔP is a maximal pressure differential between both ends of the capillary tube (in Pa) during use; andμ is a viscosity of the sample during use.
30. The test device of claim 29, wherein D is between 100.mu. and 800.mu..
31. The test device of claim 30, wherein D is between 100.mu. and 500.mu..
32. The test device of claim 23, wherein the capillary tube has at least two lengths of different internal cross sections
33. The test device of claim 23, wherein the blood sample receiving element is a blood well.
34. The test device of claim 23, further comprising a reservoir in communication with the second end of the capillary tube.
35. The test device of claim 34, wherein a reservoir in communication with the second end of the capillary tube is mounted on the support.
36. The test device of claim 23, wherein at least the blood sample receiving element, the capillary tube and a connector in electrical connection with said at least one pair of electrodes are mounted on a multi-layer support.
37. A system comprising a test device of claim 23 and of a measurement unit having at least one impedance measuring device coupled to the connecting device.
38. The system of claim 37, wherein the impedance measuring device is a multimeter.
39. The system of claim 37, further comprising a pump coupled to the second end of the capillary tube.
40. The system of claim 37, further comprising an injector adapted to add at least one agent to the blood sample during use.
41. A process comprising:obtaining a test device of claim 23;obtaining a blood sample and pumping it through the capillar tube; anddetermining a dynamic change of the value of the impedance between at least one pair of electrodes.
42. The process of claim 41, wherein the blood sample has a volume between 100 μl and 1 ml.
43. The process of claim 41, wherein the shear rate of the blood sample in the capillary tube is between 200 s-1 and 1000 s.sup.-1.
44. The process of claim 43, wherein the shear rate is between 500 s-1 and 700 s.sup.-1.
45. The process of claim 44, wherein the shear rate is 600 s.sup.-1.
46. The process of claim 41, wherein the impedance is a capacitance C.
47. The process of claim 41, further comprising adding a treatment agent to the sample in incubating the sample.
48. The process of claim 47, wherein the treatment agent is aspirin.
49. The process of claim 41, further comprising adding an aggregation agent to the sample.
50. The process of claim 49, wherein the aggregation agent is collagen.
[0001]The present invention relates to a test device for platelet
aggregation detection.
[0002]Numerous test devices for exist in the market to perform such detection. The tests are generally performed in specialized laboratories and require a long time to obtain the results.
[0003]Most known devices require isolation of platelets from the blood which needs the preparation of a sample. Examples are the trombo aggregometers BIOLA (Moscou) or EMA (Servibio) which use a laser to measure optical transmission through isolated platelets.
[0004]Another problem of most known devices is that they require a large amount of blood, for examples those using turbidimetry e.g. a device sold by COULTER (CHRONOLOG) or test devices such as BIOLA or ENTEC MEDICAL, or those using impedance such as another device sold by COULTER or the MULTIPLATE device sold by Instrumentation Laboratory. It is often necessity to dilute the blood samples with an appropriate amount of serum.
[0005]There is a need for a test device that is capable of performing routine and ambulatory test i.e. using full (or whole) blood, and preferably without being obliged to dilute the blood sample, and that is capable of giving quick results to respond to ambulatory conditions or to an emergency condition of a patient.
[0006]A step toward this aim is performed by the PFA 100 of Dade Behring which measures with full blood the occlusion time of a blood flow through an aperture in the center of a which is membrane coated with aggregating molecules such a collagen, and by the more recent device Verify Now sold by ACCUMETRICS. However, these two devices still require a large amount of blood.
[0007]DiaMed has introduced a test by colorimetry in its IMPACT device which can be performed on a small sample of blood (130 μl).
[0008]The present invention relates to a test device that is capable of performing an ambulatory test on a small sample of blood, and more particularly of full blood, and of giving a result in a relatively small amount of time, so that it is suitable for ambulatory or emergency conditions.
[0009]To that effect, the invention relates to a test device for platelet aggregation detection comprising: [0010]an element for receiving a blood sample [0011]a capillary tube connected at a first end to said element and adapted to be connected at a second end to a pressure lowering device to pump said blood sample through said capillary tube [0012]at least a pair of spaced electrodes, namely placed side by side or face to face, on the capillary tube [0013]and a connecting device to connect the electrodes to a device for measuring an impedance between said pair of spaced electrodes.
[0014]The invention also relates to a system comprised of a said test device and of a measurement unit having at least one impedance measuring device, such as a multimeter, coupled to the connecting device. The system may also comprise a pump coupled to the second end of the capillary tube and/or injection means for adding at least one agent to the blood sample.
[0015]The invention also relates to a process for using a test device as defined above, characterized in that it comprises:
[0016]a) receiving a blood sample and pumping it through the capillary tube,
[0017]b) determining a dynamic change of the value of the impedance (capacitance and/or resistance) between at least one pair of spaced electrodes.
[0018]The invention will be better understood with the following description in relation with drawings, wherein.
[0019]FIG. 1a told illustrate a first embodiment of a test device according to the invention in top view, respectively of the whole device (1a), and of its top (1b), middle (1c), and bottom (1d) layers.
[0020]FIG. 2 illustrate another multi-layered embodiment of the invention;
[0021]FIG. 3 illustrates a device 110 of the invention connected to a measurement unit 100 having a keyboard 130 and a screen 120,
[0022]FIGS. 4a to 4d illustrate a preferred embodiment for the test device according to the invention, thereof
[0023]FIGS. 5a and 5b illustrate an experimental device and the position of the electrodes of a pair
[0024]FIGS. 6 to 10, 11a, 11 b, 12a to 12c, 13a and 13b illustrate test results obtained with a test device according to FIGS. 4a and 4b. Variation ΔC (in nF) of capacitance C or normalized variation ΔCn of the capacitance is plotted versus time t in minutes or non dimensional time t/T.
[0025]The normalized variation ΔCn is obtained by the formula:
[0026]Cmax being the maximum value of the capacitance obtained when blood is circulating between the electrodes.
[0027]The test device illustrated in FIGS. 1a to 1d is comprised of a to hypodermic needle 1 connected by a resilient coupler 2 to an end 31 of a capillary tube 3 which extends along a sinuous path on a glass or plastic support 4 that may be of the size of a credit card. The hypodermic needle and the resilient coupler can be replaced by a blood well where one or more drops of blood may be deposited. The capillary extends along a sinuous or concentric path or any other geometry and has a length allowing shearing the blood sample at a chosen shear rate and time. At the other end 32 of the capillary tube 3 a vane 34 is disposed, to allow a connection of the end 32 to a closed reservoir 5 at a pressure lower than atmospheric pressure to create a pressure difference ΔP between both ends 31 and 32 of the capillary tube 3 which forms a micro channel along which a collected blood sample is sucked.
[0028]The blood sample thus circulates between ends 31 and 32 where it occupies a part of the length L of the tube 3 and is collected by the reservoir 5. A support 4 with an integrated reservoir 5 allows a self-contained operation for an in situ or ambulatory test.
[0029]For the best efficiency, it is recommended that reservoir 5 should be at the lowest pressure that would be compatible with the avoidance of degassing of the blood sample, e.g. 30 mmHg, i.e. about 4000 Pa.
[0030]In a variant, a depression or vacuum pump is connected to end 32 of the capillary tube 3, but in this case an integrated reservoir 5 is still desirable to collect the blood, because it allows discarding directly the test device with the blood sample once used.
[0031]A connector 6 is electrically coupled to pairs of metal strips 7 on the support 4 to connect to a measuring equipment pairs of electrodes 8 that are spread along the capillary tube 3, for example as shown in curved regions 35 separating linear portions 36 of the capillary tube 3.
[0032]The connector 6 allows connecting the pairs of electrodes to the equipment to measure the impedance with a set of conventional multimeters, and more particularly the value of the capacitance C between the pairs of electrodes, as will be discussed in the example below.
[0033]It is noted that only a pair of spaced electrodes 8 may be enough. However, using two or more pairs of electrodes 8 along the path of the capillary tube 3 allows obtaining several curves representing the response after subjecting the blood sample to increasing durations of shear, which brings additional information and contributes to the reliability of the test. Two pairs of electrodes may be used to determine the velocity, by determining the time span separating their transient response, and hence the shear rate in the blood sample. With more than two pairs of electrodes, the equipment may comprise only two multimeters: when the measurement starts, the first and second multimeter are coupled to the first and second pairs of electrodes. Then, when the curve representing the passage of blood before the first pair of electrodes has been recorded, the first multimeter is coupled to the third pair of electrodes, and when the curve representing the passage of blood before the second pair of electrodes has been recorded, the second multimeter is coupled to the fourth pair of electrodes and so on.
[0034]As shown in FIG. 1b-1d, the device is comprised of a sandwich of three layers. The bottom layer (FIG. 1d) is e.g.; made of plastic and comprises a first set of strips made of metal or of or conductive ink 71 having end electrodes 81, an embossed region 51 forming the bottom of the reservoir 5, and an optional spot 34 to lodge an obturator for the channel 3. The middle layer (FIG. 1c) is e.g. made of plastic and comprises an engraved channel 3 (e.g. by high speed water jet) and an engraved reservoir 5. The top layer is similar to the bottom layer (strips 72, electrodes 82 and top region 52). It also comprises an optional opening for an electronic chip 60.
[0035]As shown in the variant of FIG. 2, the support 4 may be comprised of several elements namely a bottom layer 41 having an aperture 47 forming a reservoir, a first isolating plate 42 bearing connecting strips 7, then the main plate 40 comprising the capillary tube 3 with its pairs of measurement electrodes 8 and last a top layer 43 which protects the plates 42 and 40 and may comprise a second set of connecting strips.
[0036]In the preferred embodiment of FIGS. 4a-4d, the capillary tube 3' is wound in a helical way, and has a circular, square or rectangular cross section for the circulation of blood. End 32' of the capillary tube 3' in communication with reservoir 5 (FIG. 4c) is located at the center whereas end 31' communicating with the blood well 50 is at the periphery. The volume of this blood well 50 is approximately 30 mm3 to receive a blood sample. Injection tubes 51 and/or 52 may be provided to inject an aggregation agent.
[0037]The inside diameter of the capillary tube 3' is 300μ and its length is 117 cm, to provide a shear rate of about 600s-1 to a sample of full blood with a difference of pressure ΔP=730 mm Hg (96000 Pa).
[0038]FIG. 4a shows the plate 40 in perspective, whereas figure to 4b is a top view of FIG. 4a and FIG. 4c is a cut of FIG. 2c along CC.
[0039]FIG. 4d shows a partial cut along DD and detail G thereof to show a pair of electrodes 81 and 82.
[0040]Detail G shows two electrodes 81 and 82 of a pair of electrodes placed side by side and separated by a gap d. For example each electrode occupies part of the perimeter of the tube 3' and has a length l=1 mm and width w=0.3 mm, and the electrodes 81 and 82 are separated by a distance d=1 mm.
[0041]The tests illustrated in FIGS. 6 to 13b have been performed with an experimental device provided with four pairs of spaced electrodes (811, 812; 821, 822; 831, 832; and 841, 842) that are part of a common support 70 shown in FIG. 5a, each electrode 811 . . . 842 having a length l=1 mm and a width w=0.3 mm (see FIG. 5b) and occupying half of the circumference of a capillary tube having an internal diameter of D=300μ. The pairs of electrodes are longitudinally spaced by a distance d=1 mm. A supple capillary tube 71 having a chosen length L is disposed to follow a sinuous path and is in contact with the successive pairs of electrodes.
[0042]FIG. 6 illustrates the change AC in the value of the capacitance with two neutral liquids, namely distilled water and a coloured solution, for three pairs of electrodes.
[0043]The passage of the front separating the two liquids produces a sudden variation of the value of the capacitance C. Signals at three pairs of electrodes 1,3 and 4 have been shown.
[0044]FIG. 7 illustrates the change AC with a flow of distilled water followed by 50% diluted blood with an aggregating agent (collagen at 3 μg/ml), with four pairs of electrodes.
[0045]FIG. 8 illustrates a flow of diluted blood that has been incubated during 15 nm with a solution of aspirin at a concentration (5 mM) that is known to inhibit platelet aggregation to which collagen (3 μg/ml) has been added after incubation with signals from three pairs of electrodes 1,2 and 3.
[0046]For this sample, aspirin is efficient to prevent aggregation since the change of ΔC exhibits a sudden variation.
[0047]FIG. 9 illustrates a flow of blood that has been incubated (curve I) during 15 mm with aspirin (5 mM) or not incubated (curve II). It can be seen that curve II (no aspirin added) doesn't show the sudden change of curve I, indicating that platelet aggregation may have occurred in curve II, not in curve I.
[0048]FIG. 10 adresses the specificity of the test with a sample of blood without platelets.
[0049]The passage of the sample produces a fast rise ΔC of the value of the capacitance C, which confirms that when a slow transition is present, it is specific of platelets.
[0050]FIGS. 11a and 11b test the influence of an aggregating agent (collagen) on a sample of diluted blood.
[0051]FIG. 11a shows curves obtained with a sample without addition of collagen and FIG. 11b shows curves with a sample of the same blood with an addition of collagen (3 μg/ml).
[0052]This comparison shows that the main effect is obtained by the shear rate on the sample (around 600 s-1 in the example) and that the addition of an aggregating agent may not be necessary. The test device according to the invention may thus be used with a sample of full (or diluted) blood with or without addition of an aggregating agent such as collagen.
[0053]FIGS. 12a to 12c show the effect of a treatment with aspirin. FIG. 9a shows the curve before the treatment. FIGS. 9b and 9c show curves after 3 days of treatment (500 mg/day) respectively without addition of collagen to the sample, and with addition of collagen (3 μg/ml). The sharp change of ΔC in FIGS. 9b and 9c indicates an efficiency of the treatment, independently of the presence of collagen.
[0054]FIG. 13a and FIG. 13b show the correspondence between a test with 50% diluted blood (FIG. 13a) and a test with full blood (FIG. 13b). These curves show that the results are consistent and that the test device may be used either with full blood or with diluted blood.
[0055]Tests on full blood are of course preferable for ambulatory or emergency tests. But tests on diluted are also of interest for various reasons, especially since the dilution allows obtaining lower shear rates with the same device.
[0056]Another way of obtaining different values of shear rates is providing the capillary tube with two or more lengths of different cross-section. Pressure measurement at the beginning and at the end of each section allows to determine the value of the shear rate in each cross-section.
[0057]In the test device, the ratio L/D between the length L and an internal diameter D of a circular capillary tube and S being the internal cross-section of a capillary tube:
L / D = L 2 π / S ≦ Δ P 4 γ w μ ##EQU00001## [0058]γW being the minimal shear rate in s-1, nominally 200 s-1 [0059]ΔP being the maximal pressure differential between both ends of the capillar tube (in Pa), nominally 105 Pa [0060]μ viscosity of the sample (nominally 0.003 Pas for a blood sample).
[0061]The shear rate of the blood sample in the capillary tube is for example between 200 s-1 and 1000 s-1, more particularly between 500 s-1 and 700 s-1, the preferred value being 600 s-1. Note that the phenomenon of platelet aggregation depends on the shear rate and the above ranges are pertinent. D is for example between 100 μm and 800 μm and preferably between 100 μm and 500 μm.
[0062]The volume of the blood sample is for example between 15 μl and 1 ml (e.g. 100 μl) and it is for example chosen so that the sample occupies only part of the length of the capillary tube.
[0063]The maximal transit time in the capillary tube is about 3 minutes. The above formula and the following table allow to adapt the shear rate, the pressure differential ΔP and the transit time in a capillary tube of length L. The following table shows for different values of the internal diameter D (mm) of the capillary tube and a shear rate of 600 s-1 the speed S (mm/s), the flow F (μl/s) and the maximal length Lmax (m) of the capillary tube for ΔP=105 Pa, and the corresponding volume V (μl) of the capillary tube, as well as the Reynolds number R for a viscosity of 3 mPas and a volumetric mass of 103 kg/m3
TABLE-US-00001 D (mm) 0.2 0.4 0.6 0.8 1 S (mm/s) 15 30 45 60 75 F (μl/s) 0.47 3.77 12.72 30.16 58.9 Lmax (m) 2.8 5.6 8.4 11.2 14 V (μl) 88 704 2380 5600 11000 R 1 4 9 16 25
[0064]The test device according to the invention is compatible with a certain degree of miniaturization. Samples of volume lower than 100 μl (e.g. . . . 15 μl which corresponds to a single drop of blood) can be accommodated. Also, sub-millimetre electrodes may be used. L, which is equal to 1 mm in the examples, may be reduced, e.g. to 120 μm, down to about 25 μm.
Patent applications by CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
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