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Painters' Paintings at the National Gallery

When Henri Matisse bought a small painting by Paul Czanne, Three Bathers, in Paris in 1899, it was far beyond his means. But he did everything he could to buy it his wife Amlie even pawned a cherished emerald ring. Once acquired, the Czanne became a fixture of Matisse's studio for 37 years. It was the first thing he would look at when he woke up. It changed his life and work. He later said: "It has supported me morally at critical moments in my venture as an artist; I have drawn from it my faith and my perseverance."

This magnificent painting is among the masterpieces in this show of works Hermes enamel necklace copy once owned by artists, from Anthony Van Dyck through Joshua Reynolds to Edgar Degas and Lucian Freud. I doubt any painting was as loved by its owner as Matisse's Czanne. The pictures collected by these artists were far more than just objects of affection and moral support. They used them to educate themselves and others, to pit themselves against their heroes and peers, to show off their connoisseurship, to decorate their show homes.

Painters' Paintings offers the tantalising prospect of a glimpse behind the artist's vision. The show's curators encourage this by showing paintings made by the artist collectors alongside works they gathered. A self portrait accompanies each one.

For this approach to work, though, the comparisons need to be top notch. The picture that prompted the show in the first place is Jean Baptiste Camille Corot's Italian Woman, a work left by Lucian Freud to the National Gallery on his death in 2011. It's a beautiful little picture, marked by the monumentality yet delicacy of the figure and by Corot's loose handling of paint, qualities shared by another picture Freud owned, John Constable's Portrait of Laura Moubray.

Yet the Freud portraits here neither illuminate his response to his possessions nor show him in a good light amid his heroes. An etching that appears to have little to do with the Corot is offered as a spurious comparison, and a self portrait from 2002 is Freud at his least fluent, its surface intense but horribly overworked, the paint limp and dry.

Freud also owned a small early painting by Czanne. His interest in this erotic scene which features nude figures in what might be a brothel amid dishevelled sheets, with a maid bringing in tea is understandable. He was attracted not just by its eroticism but by its awkwardness: he found it funny.

And it is an odd picture, with so much flaccid flesh. Freud even made a huge painting in response to it. Sadly, that work is only shown as a black and white reproduction on a label. Instead, as a comparison we have another minor Freud nude evoking the Czanne, its surface again overworked, a web of thick pentimenti undermining the final image. The Czanne might be awkward in its forms but look closely at the delicacy of the Frenchman's handling, how he built up the surface in dabs and patches that belie the crudeness of the drawing. I suspect Freud would have found this wondrous, given how hard won his own paintings always were.

If the section devoted to Freud is a flawed opening (the show works backwards, chronologically), the three rooms that follow, one dedicated to Matisse and two to Degas, are the show's high points. As well as the startling Czanne bathers, Matisse owned one of Czanne's portraits of his wife, a spellbinding Gauguin and the great Degas painting Combing the Hair, now a favourite in the National Gallery. These paintings possess, in different quantities, so much of what is best in Matisse: high colour and pure harmony, reduction and order, boldness in composition. There are also two of Matisse's Picassos here including a brilliantly dark portrait of Dora Maar that Matisse kept in his Nice hotel room: a spur, a challenge by his most illustrious peer.

Degas bought works by his contemporaries and followers in vast numbers, as well as numerous works by the artists of the generations immediately before him Delacroix, Ingres and Corot among them. Degas gave one floor of his Paris house over to his collection, the paintings standing on a forest of easels.

The Degas rooms capture this voracious collecting and are a thrilling exhibition in themselves. The works evoke his position as a painter between the avant garde and the academy, between classical order and impressionist rupture.

Middling works by minor painters only enhance our appreciation of Degas's own: Georges Jeanniot's Conscripts from 1894 is shown next to Degas's Young Spartans, painted 30 years earlier. The Degas teems with light and colour, energy and ambition; the Jeanniot is leaden by comparison.

Transcendent pairings abound: a Delacroix sky study next to Degas's own; a Corot capturing the Roman campagna and Degas's painting of the Bay of Naples.

After the heights copy cartier earrings of such an inspired artist collector as Degas, the show sags. The problem is that it is too bound by the works in the National's collection. It managed to buy numerous works from the sales of Degas's collection after his death, hence the richness of that section. But it means the rooms that follow feel inadequate. Turner is evoking a sense of loss, lamenting the decline of Britain's naval power. Messages aside, the painting itself is a gorgeous contrast of styles: details of the ship are finely done, the rigging lightly sketched, the paddles of the boat slicing the water, while the sky is a glorious explosion of colour, thick layers of Hermes replica necklace oil paint, the sun and sky by Hermes necklace knock off turns silver and gold, red and purple. In 2005, it was chosen as the nation's favourite painting in a Radio 4 poll.

Self Portrait with Two Circles, Rembrandt, c. At the time of painting, he'd lost his wife, three of his children, and his mistress. He sits upright, square jawed, jaw set defiantly, if a little sourly it's a long way from his early, energetic, playful self portraits. Four years later, after the loss of another son, he would by dead, having killed himself. The painting is a large one: give it some time. It's a peculiar experience to stand in front of it and sympathise with a stranger. It's said van Gogh was spurred to paint the flowers in order to have new work to impress Paul Gauguin, whom he planned to share a studio with. On a diet of coffee and booze, he painted the first four in just six days. While visiting the Sunflowers, be sure to see his famous Chair; A Wheatfield, with Cypresses; and the magnetic 1899 painting Two Crabs, which sometimes, sadly, goes overlooked. Too ill to paint, Matisse invented a new form with the cut outs and the later Blue Nude series, as seen here, are deceptive: they seem simple, but the limbs swirl around each other, and Matisse captures something very human in the posture particularly noticeable here in the slight incline of the neck. Straightforward, but captivating.

Venus, Sandro Botticelli, c. It contains the same ground breaking nudity Christian influence had put a bit of a damper on that for a few centuries beforehand and again, it's all in the eyes here: the expression could mean one of a thousand things. Afterwards, head to the National to see Venus and Mars, from a few years earlier.

Volker H. Schneider / Gemldegalerie, SMB / Jrg P. It's partly famous as the barmaid's reflection has puzzled onlookers since it was first shown: still, Manet's last major work proved to be a hit, and so it has remained.
Piaggio Porter Electric MPV

I have to admit that I wasn't particularly excited at the prospect of testing the Piaggio Porter Electric. All I knew was that it was a small, electrically powered van with a top speed scarcely higher than the 30mph limit. How much fun was that going to be?

By the time it turned up a couple of weeks ago, things were looking different.

The price of petrol was breaking through the 1 per litre barrier, and the fuel protesters were on Hermes replica necklace the television news bulletins threatening to bring the country to a halt. Suddenly, any vehicle that could be charged up from the mains was starting to look pretty attractive.

My feelings towards the Piaggio remained warm throughout our test. But in order to appreciate electric vehicles, you have to understand the limitations and characteristics of the type. The 35mph top speed and maximum 85 mile range seem, on the face of it, to be restrictions that would be almost impossible to live with. However, in town, 35mph is enough. Electric vehicles provide a lot of torque at low revs; if anything, take off from rest is rather abrupt. Driving is otherwise extremely fake Hermes necklace easy. A small lever between the front seats is moved forwards or backwards, depending on the direction of travel. There is no clutch or gear lever you just press the accelerator to go and the brake to stop.

It takes eight hours to recharge the Piaggio, which can be a bit awkward to organise. Nobody wants to leave a downstairs window open overnight, and most letter flaps were not designed to have plugs posted through them. A catflap would work best, or a space in your garage with access to a wall socket.

The Piaggio did once run out of puff on a near full charge, after one of our Cambridgeshire readers ran it at high speed uphill on the Ely bypass (" high speed" and "uphill" being relative concepts in the cases of the Porter Electric and East Anglia respectively). After a short push and a rest, it was able to continue as normal.

One obstacle to the take up of electric vehicles is the high initial price. Whether the sums add up depends on your pattern of driving and the value you place on reducing local pollution; the power station that produces the juice for your electric car is probably on someone else's doorstep, not yours. Electric vehicles are exempt from road tax and the London congestion charge.

In all, the electric experience was a surprisingly enjoyable and interesting one. If, in order to save the planet, we all end up driving Piaggio Porter Electrics, it certainly won't be the end of the world.

Mark Tyler, 35, general practitioner from March, Cambridgeshire, and Emma Tyler, 6,


"The driving position was so high that your view through the windscreen was obscured by the rear view mirror and roofline. I was stooping to get a clear view of the road. The clearance between the front seat base and the A pillar was very narrow. The ride itself was very reasonable, but the absence of power steering made for hard work."

Emma says: "It was a quiet comfortable ride with a good view through the big windows. I though it was too high for small children to get in and out of easily."

Mick King, 50, paramedic from Warboys, Cambridgeshire


My initial impressions were very poor and in keeping with most electric vehicles why can't we have better styling on battery powered vehicles? The start up procedure is simple, with one lever for forward/reverse drive (no gearbox) and the vehicle is driven almost like an automatic car Hermes imitation necklace however, this is where the similarity ends! On inclines the Piaggio can be held on the brake but once this is released the vehicle has a tendency to roll back before the throttle picks up. It can be held on the handbrake but this emits an ear piercing warning not ideal. Overall the driving position was excellent and comfortable, but the drive itself is very underpowered.

Derek Morris, 45, consultant from Sawston, Cambridgeshire


Driving it around town was fine I didn't like the accelerator pedal, though, which had an "on off" quality. The suspension was a bit stiff so there was some harshness over speed bumps. Acceleration and speed were sufficient around town but out on the open road you quickly begin to hold up the traffic even at the indicated 40mph or so that seemed to be its top speed. It was surprisingly roomy inside and the driving position was fine despite being bus like, although the unassisted steering was a little heavy. The interior trim, although it may be normal for a small van, wasn't really up to the price; I was amazed there was no radio, for example. For most cars, participants must be over 26 and have a clean licence.

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Portable Car Air Conditioner

The most popularly installed air conditioner (AC) is the window type, which is applicable for cooling or heating temperature inside an enclosed structure. It is a fixed unit and is not meant for carrying from one area to another. Considering this drawback, portable air conditioners are configured so Hermes Kelly bag knock off that they can be used for creating a suitable environment in temperature sensitive areas, especially during extreme climatic conditions.

A portable air conditioner is basically designed for use in non standard spaces, such as boats, warehouses, vehicles, and recreational camps. In other words, this appliance is a Hermes handbag copy real buy for controlling temperature in small, closed or open areas, which are not suited for central air conditioning or fixation of regular window air conditioners. With a portable air conditioner, you can enjoy an outing to the fullest even in extreme weathers. It is also preferred for cars that have no air conditioning system. While choosing it, look for the model that controls temperature quickly at the expense of low energy.

Based on the machinery, there are two basic types of portable air conditioners, namely, evaporative technology and refrigerant technology. The evaporative portable air conditioner is preferable for use in cars, while the refrigerant type is mostly applied Hermes Birkin bag replica for operation on room floors. The evaporative configuration is more energy efficient in comparison to the refrigerant type. Refrigerant technology is more or less similar to window AC, regarding the size and coolant usage. As far as the working of portable air conditioner for cars is concerned, a water reservoir is used for relocation of heat, which then works by cooling and humidifying the air inside the car. This model is usually operated by using a 12 volt power converter. For maintenance handbag Hermes copy of this small car AC, the water reservoir should be refilled whenever required. A purifying filter, as the name indicates, is installed for cleaning the air inside the car and generating healthy air for breathing. The dehumidifier removes excess moisture from inside the car. The lowest price, portable AC for vehicles starts from USD 50. And depending upon the brand and added features, this spot cooling device may cost as high as USD 400 to USD 600 (or even more). To get a good product at an affordable price, always conduct a brief research about the various brands prior to buying this portable unit. Also, check the reliability, energy efficiency, compactness, installation, maintenance, and warranty of the AC before purchasing.

A portable car air conditioner is very convenient to use as it does not vibrate or generate any noise, which is not so in case of window models. One of the major advantages of this cooling appliance is that when you are not driving the vehicle, you can make use of it at home while watching TV or sleeping. Since it is lightweight, you can move it wherever cooling and/or freshening is needed. Hence, if you have a portable air conditioner, fixation of cooling systems in every room may not be required, thus saving the cost of buying and installing them.
Queens Boulevard is Going Down a New Path

Officials say the Queens Boulevard Vision Zero project will make it a safer place for pedestrians and cyclists. But some residents have concerns about the plan. NY1's Clodagh McGowan has the story.

Cyclists now have a clear bike route along Queens Boulevard from Roosevelt Avenue to 73rd Street in these protected bike lanes. It's part of the first phase of the Department of Transportation's $100 million overhaul to make Queens Boulevard Hermes handbag imitation safer.

According to the DOT, this stretch saw the highest concentration of fatalities between 2009 and 2013, with six people killed and an additional 36 severely injured.

Elaina Tolson says she was hit by a bike while crossing Queens Boulevard recently. She hopes cyclists will take caution.

"They ride without helmets, they ride without reflective gear. They take chances as it is, I Hermes bag copy don't think it's necessarily the best idea. Hermes Kelly handbag copy It's pretty risky," said Tolson.

While some Woodside residents say they're happy to see protected bike lanes for cyclists on the boulevard, that doesn't mean they'd feel safe riding in it.

"Not now. It's too tight," said one resident.

But Councilman Jimmy Van Bramer is not afraid to take a ride on what has been dubbed the Boulevard of Death.

"It's proven that when we have bike necklace Hermes replica lanes, when we have protected bike lanes, when we have an expanded bike network, when we have complete streets, fewer people die," said Van Bramer.

Cyclists say having a clear division between the lanes is a great plan.

"I feel way protected from cars. I only wear this because of cars. Not because of pedestrians, not because of other bikers," said one cyclist.

In addition to the bike lanes, some intersections no longer allow cars to cut all the way across the boulevard. It's causing frustration for drivers who say main thoroughfares, like 58th Street, are getting backed up.

"It's too much traffic," said one driver.

A DOT spokesperson tells NY1 the street closures gives pedestrians more opportunities to safety cross the street by connecting the medians. The first project is expected to be complete by October.
Photo gallery of CHEK

Victoria Local heroes, celebrities and others were on the streets of Victoria selling the Times Colonist to raise funds for children's and family literacy programs as part of Raise a Reader. CHEK televison personalities left Gordie Tupper, Dana Hutchings and Jennifer Crosby on Douglas St. June 9, 2009.

Scott Fee and Jennifer Crosby on the news set in March 2007.

The annual CHEK News Christmas Charity Drive Thru helped hundreds of needy families in the Greater Victoria area.

An all candidates meeting in 2006 at CH studios with Liberal Keith Martin, Conservative Gary Lunn , NDP Jean Crowder and Green Party Andrew Lewis, with host CH News Anchor Ed Watson .

Some of the fake hermes H bracelet pink mebers of the CH news team relax in the news studio in 2004 before heading out for their island road trip this weekend. (L2R) Ed Bain, Hudson Mack, Stacy Ross, Bruce McAllister, Veronica Cooper, and Meribeth Burton.

Veteran reporter Harry Maunu on his last day of work in 2000.

1997 CHEK anchors Hudson Mack and Jill Krop.

Michaela Pereira and Gordie Tupper, hosts of CHEK Around, 1997.

Ida Clarkson at CHEK TV in 1982.

New CHEK Television facility on Kings Road, Victoria (1984).

Left to right: Jim Nicholl, CHEK production manager, Lynn Charman, CHEK director, Mike Hill, architect with the firm of Howard/Yano, Terry Lineham, CHEK Collier de Chien hermes bracelet copy building manager and Roy Gardner, CHEK general manager.

The 1976 launch of a new album featuring Vancouver Island artists. Holding copy of the album are, on left Bruce Payne, host of Daybreak program on CHEK TV. Stan Cayer, a Vancouver promoter and head of SGM Records. Gathered around Payne and Cayer are, from left, Jackie Schille of Victoria, Al Cottell of Nanaimo, Linda Delmer of Nanaimo, Tom Gough of Victoria, Glenda Graham of Nanaimo (back), Sherry Smith of Nanaimo (between Payne and Cayer), Barry Worth of hermes mens bracelet replica Duncan and Larry Friesen fake hermes h bracelet Knockoff of Duncan, all featured on the record.
Rapid detection of single bacteria in unprocessed blood using Integrated Comprehensive Droplet Digital Detection

Rapid detection of single bacteria in unprocessed blood using Integrated Comprehensive Droplet Digital DetectionDong Ku Kang1, 2, 3, 4, 5 n1, M. Monsur Ali1, 2, 3, 4, 5 n1, Kaixiang Zhang1, 2, 3, 4, 5, 6, Susan S. Huang7, Ellena Peterson8, Michelle A. Digman5, 9, 10, Enrico Gratton5, 9 Weian Zhao1, 2, 3, 4, 5Nature Communications 5, Article number: 5427 (2014)doi:10.1038/ncomms6427Download CitationBacterial infectionBacterial techniques and applicationsMedical and clinical diagnosticsSepsisAbstractBlood stream infection or sepsis is a major health problem worldwide, with extremely high mortality, which is partly due to the inability to rapidly detect and identify bacteria in the early stages of infection. Here we present a new technology termed 'Integrated Comprehensive Droplet Digital Detection' (IC 3D) that can selectively detect bacteria directly from milliliters of diluted blood at single cell sensitivity in a one step, culture and amplification free process within 1.5 4h. The IC 3D integrates real time, DNAzyme based sensors, droplet microencapsulation and a high throughput 3D particle counter system. Using Escherichia coli as a target, we demonstrate that the IC 3D can provide absolute quantification of both stock and clinical isolates of E. coli in spiked blood within a broad range of extremely low concentration from 1 to 10,000 bacteria per ml with exceptional robustness and limit of detection in the single digit regime.IntroductionBlood stream infections (BSIs) are a major cause of morbidity and mortality. Sepsis resulting from a BSI annually affects over 18 million people worldwide and 700,000 in the United States, with a mortality rate of 30 40% (refs 1, 2) Sepsis and other aggressive bacterial infections associated with BSIs are often times managed within intensive care units with associated high costs, which impose significant health care, economic and social burdens. For instance, each septic patient in the United States incurs costs of approximately $25,000 during hospitalization, corresponding to $17 billion annually1,2. The extremely high mortality of blood infections is due, in part, to the inability to rapidly detect, identify and thus treat patients with appropriate antibiotics in the early stages of infection1,2,3. The initial treatment with empirical broad spectrum antibiotics not only is inadequate but also encourages antibiotic resistance4,5. It is widely recognized that effective detection and monitoring of patients to diagnose a BSI at an early stage have a profound effect on survival rates1,2,3. However, the present gold standard, bacterial blood cultures coupled with susceptibility testing for drug resistance, requires days to obtain a result. Recent amplification based molecular diagnosis methods including PCR can reduce the assay time to hours but are often not sensitive enough to detect bacteria that occur at low concentrations in blood (1 100 colony forming unit (CFU) per ml) as is commonly found in adults with BSI and therefore often still require a culture enrichment step6,7. Moreover, these conventional methods typically suffer from poor specificity and high background signal because a target bacteria is surrounded by billions of non target species (for example, red blood cells) in the blood sample. More recent nano hermes bracelet sale replica and micro systems including droplet microfluidics (for example, digital PCR)8,9,10,11,12,13,14,15,16,17,18 can improve detection sensitivity and selectivity but typically are limited to microlitre sample volume, which cannot handle the required clinical sample volume (millilitres) and throughput. Inevitably, the existing methods typically require expensive equipment and lengthy, complex sample processing (for example, cell lysis, nucleic acid extraction, centrifugation, magnetic separation, washing and signal amplification) for target purification and enrichment, which not only results in significant loss of rare target organisms, and therefore contributes to a high false negative rate, but also limit their widespread use especially in a point of care setting (for example, in an ambulance)6.We present herein a platform technology called Integrated Comprehensive Droplet Digital Detection (IC 3D) that is able to selectively detect bacteria from millilitres of unprocessed (although diluted) blood at single cell sensitivity in a one step, and culture and amplification free reaction within 1.5 4h.ResultsOverview of IC 3DOur IC 3D system integrates real time DNAzyme sensor technology, droplet microfluidics and a high throughput 3D particle counter system (Fig. 1). DNAzyme sensors used here are short catalytic oligonucleotides that are identified by in vitro evolution to specifically react with the lysates of target bacteria, leading to a rapid fluorescence signal19. Specifically, blood samples are mixed with the DNAzyme sensor solution including bacteria lysis buffer within a microfluidic channel, which is then immediately encapsulated into 100s of millions of individual picolitre droplets. The confinement of bacteria in droplets that serve as 'microreactors' significantly increases (i) the concentration of released target molecules in the droplets that contain bacteria such that single bacteria can be detected by the DNAzyme sensors in a rapid manner and (ii) target/background ratio to minimize interference from nonspecific binding and noise. In the IC 3D system, droplets are collected following generation into a vial and imitation hermes bracelet sale analysed using a high throughput 3D particle counting system. The 3D particle counter was recently developed by Gratton and colleagues and can robustly and accurately detect single fluorescent particles from millilitre volumes within minutes20,21. In the IC 3D system, the compartmentalized, target specific reaction mediated by DNAzyme sensors is critical to 'light up' the droplets that contain target bacteria so that they can be detected by the 3D particle counter with exceptionally high reliability and clinically relevant throughput.Figure 1: Schematic description of the IC 3D technology.(a) Blood samples and DNAzyme sensors are mixed and then encapsulated in 100s of millions of micrometre sized droplets. DNAzyme sensors produce an instantaneous signal in the droplets that contain the bacterium. (b) In the IC 3D, droplets are collected and analysed using our high throughput 3D particle counter that permits accurate detection of single fluorescent droplets in a several millilitre pool of non fluorescent droplets within minutes.DNAzyme sensors selectively detect target bacteria in bulkDNAzymes (also called 'DNA enzymes' or 'deoxyribozymes') are synthetic single stranded DNA oligonucleotides with catalytic activities22,23. These catalytic DNA molecules are generated in vitro from a vast random library using a combinatorial approach called in vitro evolution24,25. One of the most established DNAzymes is a RNA cleaving DNA motif that can cleave a DNA RNA chimeric substrate at a single ribonucleotide junction22,26. Ali and colleagues have recently harnessed this unique property to generate bacteria specific DNAzyme sensors through in vitro evolution of a vast DNA library against crude extracellular matrix (CEM) components of target bacteria as complex targets19. A later study indicated that the cell lysate produces much higher fluorescence signal compared with the CEM by the DNAzyme sensors, suggesting that the cells contain higher amount of targets than CEM27. In our study, therefore, we decided to lyse the cells to facilitate detection and first tested a series of detergents and enzymes to optimize the cell lysis condition and identified that lysozyme most efficiently lyses bacteria without interfering with droplet generation and stability or DNAzyme sensor function (see Methods section for details).We sought to use these rapid, fluorogenic DNAzyme sensors in our IC 3D system. As shown in Fig. 2a, the sensor contains a DNAzyme domain that is ligated with the DNA RNA chimeric substrate where the ribonucleotide cleavage site is flanked by a fluorophore and a quencher. This 'inactive' state has a minimal fluorescence signal because of the close proximity of the fluorophore and the quencher. In the presence of target bacteria, E. coli used herein as a model system, DNAzymes will bind to target molecules produced by bacteria and cleave the substrate. The cleavage event (Supplementary Fig. 1) frees the fluorophore from its quencher, thereby generating a high fluorescence signal (Fig. 2b). Moreover, the DNAzyme sensor is able to distinguish target E. coli from control bacteria or mammalian cells with high selectivity (Fig. 2c). We further demonstrate that the DNAzyme sensors previously isolated using stock isolates of E. coli19 can robustly and selectively detect clinical E. coli isolates that were spiked and then lysed in blood (Fig. 2d). It is interesting to note that although the DNAzyme sensor can detect all clinical E. coli samples, the fluorescence intensity varies between samples, which might reflect the potential molecular heterogeneity between different E. coli strains. Furthermore, fluorescence intensity appeared to be slightly increased in the C. freundii groups comparing to the blood alone control but it was statistically significantly less than the E. coli groups (P0.001, two tailed Student's t test) and was not statistically different than other clinical control isolates (P=0.85, two tailed Student's t test). Nonetheless, this points out the need to rigorously validate the sensor specificity using larger number of patient samples in the future. This also suggests that by including appropriate positive and negative selection targets in the in vitro evolution process, it is feasible to generate DNAzyme sensors that can distinguish different strains of the same bacterium species, which will hermes gold bracelet fake need to be validated in the future work. As our goal is to develop a 'mix and read' assay that uses whole blood with no or minimal sample processing (for example, dilution), we further tested the sensor performance in blood and found that our Fluorescein/Dabcyl modified DNAzyme sensors produced a sufficiently high fluorescence signal in response to E. coli spiked in blood that was diluted by sensor solution to various volume ratios (Supplementary Fig. 2a) with a 10% final blood concentration determined to be optimal and therefore used in subsequent droplet experiments (below). We hypothesize that optimization of dye pairs, especially using near infrared dyes that are less interfered with by blood autofluorescence in the future, can further improve sensor performance (for example, signal/noise ratio) in blood. We further hermes enamel bracelet Knockoff demonstrated that the DNAzyme sensors exhibited sufficient stability in blood within the time frame (1.5 4h), we target for future clinical use (Supplementary Fig. 2b). If necessary, the use of well established chemically modified nucleotides (for example, phosphorothioates) in DNAzymes or RNase inhibitor in the assay buffer can further increase sensor stability in blood.Figure 2: Real time DNAzyme sensors selectively and rapidly detect target E. coli lysates in bulk.(a) Proposed mechanism of how the DNAzyme sensor generates a fluorescent signal upon interaction with the target. The target(s) produced by the bacterium binds to the inactive DNAzyme sequence (red), which undergoes a conformational change to activate the DNAzyme. The activated DNAzyme catalyses the cleavage of the fluorogenic substrate at the ribonucleotide junction (R), leading to the separation of the fluorophore (F) and the quencher (Q) to produce a high fluorescence signal. (b) DNAzyme sensor produces real time fluorescence signal in the presence of the target E. coli K12 lysates. By contrast, a mutated DNAzyme sequence is inactive. Lysates from 10,000 bacteria and 50nM DNAzyme were mixed in a 50 l final volume in HEPES buffer and signal was recorded using a fluorescence plate reader. (c) DNAzyme sensor specifically detects E. coli strains but not non target bacteria or mammalian cell human T cell lymphoblast CCRF CEM and human umbilical vein endothelial cells (HUVECs). Lysates from 10,000 cells and 50nM DNAzyme were mixed in a 50 l final volume in HEPES buffer and incubated for 30min. DNAzyme reaction products were analysed by polyacrylamide gel electrophoresis. The percentage cleavage for each reaction was derived, normalized against DNAzyme alone control and presented as 'Relative fluorescence'. (d) DNAzyme sensors can selectively detect clinical E. coli isolates. Bacteria (1,000CFU) isolated from 11 different patient samples were incubated with 100nM DNAzyme and 1mgml1 lysozyme in 10% of blood for 30min. Fluorescence intensity was obtained using a fluorescence plate reader, normally against DNAzyme alone control (con) and presented as 'Relative fluorescence'. Data are obtained in a single blind experiment. In c and d, all experiments were performed in triplicate.DNAzyme sensors rapidly detect single bacteria in dropletsWe next integrated DNAzyme sensors with the droplet microfluidics system to test our hypothesis that the confinement of bacteria in droplets can significantly increase the sensitivity (that is, single cell) and shorten the detection time. The 'droplet microfluidics' system enables the generation and manipulation of monodisperse, picolitre sized (typically 5 to 100m in diameter) liquid droplets in an immiscible carrier oil fluid (that is, water in oil emulsion)28,29,30,31,32,33. Droplet microfluidics is an emerging platform for ultra sensitive biological detection and analysis. In particular, the pioneering work from the Weitz and Griffiths groups and from companies including Bio Rad and Raindance have demonstrated a range of droplet based 'digital' assays for nucleic acids (for example, digital PCR), cells and organisms10,31,34. However, the traditional droplet system is limited by its low throughput in droplet analysis and therefore not amenable for analysis of large volume samples (see the 'Rapid detection of single bacteria in clinical blood samples' section)31,34.In our study, droplet microfluidics were fabricated using standard soft lithography and operated following previously established procedures (see 'Fabrication of droplet based microfluidic device' in Methods)35. As illustrated in Fig. 3a, the poly(dimethylsiloxane) (PDMS) chip has one oil inlet and two aqueous inlets (one for bacteria containing buffer or blood with the other one for the DNAzyme sensor mixed with bacterial lysis reagent (lysozyme)). Uniform picolitre sized droplets were generated using standard syringe pumps at a rate of approximately 2,000Hz by flow focusing of the resulting stream with HFE 7500 fluorinated oil containing 1.8% (w/w) perfluorinated polyethers with polyethyleneglycol surfactant (Fig. 3b and Supplementary Movie). We generated droplets with different sizes ranging from 5 to 50m in diameter simply by tuning the microfluidic channel size and flow rate. Figure 3c shows a representative image of 30m droplets that contain 10% blood. We found that these droplets can be stably stored without leaking or merging for months at room temperature even when closely packed in a vial. High throughput droplet generation can be achieved using a multi layer microfluidic device that contains multiple, parallel droplet generating structures (see 'Fabrication of droplet based microfluidic device' in Methods and Supplementary Fig. 3 for an eight channel device). To achieve clinically useful throughput, we are currently developing a device that contains 256 droplet generating channels, which is able to convert 1ml blood to 25m droplets containing 10% blood at a generation rate of 2,000Hz per channel in 40min. In addition, the use of larger droplet and smaller blood dilution factor can further significantly reduce the droplet generation time.Figure 3: Workflow of microencapsulation.(a) Layout of the droplet based microfluidic device. Devices were designed with three inlets; one for oil and the other two for blood samples and DNAzyme/bacterial lysis buffer. (b,c) Representative microscopy images showing uniform 30m droplets containing 10% blood and sensor solution are being generated using flow focusing. Scale bar, 200m. In c, blood contents especially red blood cells are clearly visible in droplets. (d) Droplets collected in the cuvette used for 3D particle counter experiments. (e) Representative fluorescence microscope images demonstrate DNAzyme sensors (250nM) light up the droplets that contain single E. coli K12 in 10% blood after 3 h reaction. Left panel: overlay of fluorescence and brightfield. Right panel: fluorescence. Scale bar, 200m.As in adult BSI, bacteria are in low numbers in patient's blood (typically 1 100CFUml1), when encapsulated in picolitre droplets, each droplet will contain one or no bacterium. Therefore, it is critical to test if our system is able to detect a single bacterium in a droplet. Specifically, bacteria were statistically diluted to achieve a range of concentrations from 10 to 107ml1 spiked in HEPES buffer or blood and compartmentalized in droplets. In some experiments, bacteria were stained with Syto17 (red), which allows us to co localize the bacterium with the DNAzyme sensor signal (green) in the same droplet to determine false positive and negative rates. In this section, fluorescent droplets were imaged by conventional fluorescent (or confocal) microscopy or counted on chip in a 1D microfluidic channel at a throughput of 200 droplets per s using a custom built confocal microscope equipped with APD detectors (see '1D on chip detection system' in Supplementary Methods). We first demonstrate that, in buffer, the DNAzyme sensor system is able to detect single target E. coli K12 that is lysed in a droplet (5m in diameter) within 8min (Supplementary Fig. 4). This pilot experiment tested our hypothesis that the confinement of bacteria in droplets enables single cell sensitivity and reduced detection time. For detection of bacteria in blood, we needed to optimize droplet size: although smaller droplet sizes lead to higher target concentrations from single cells (which would increase the signal/background ratio and decrease the detection time), it is technically challenging to encapsulate blood contents including red and white blood cells into too small sized droplets. We determined that droplets 25m in diameter are optimal for this purpose and therefore used for subsequent blood droplet experiments. Using fluorescent microscopy (Fig. 3e) or 1D on chip droplet counting system (Supplementary Fig. 5), our system is able to selectively detect single target E. coli K12 in 10% blood in droplets. Furthermore, by co localizing with the Syto17 signal, we observe that our encapsulated DNAzyme sensor system possesses zero false positive rate and minimal false negative rate (0.5%) from 70,000 droplet counts in triplicate experiments, which we performed using E. coli K12 as positive target and sensor alone or control bacteria as negative controls (Supplementary Fig. 5). Finally, although the DNAzyme sensor signal generation in a 25 m blood droplet is not as rapid as that in a 5 m buffer droplet, a measurable fluorescence signal can be observed within 3h in response to a single bacterium in blood (Fig. 3e and Supplementary Fig. 5; see below for further DNAzyme kinetics study).Rapid detection of single bacteria in clinical blood samplesWe have demonstrated above that the encapsulated DNAzyme sensor system can detect target bacteria in a rapid manner with single cell sensitivity in the droplet. However, the 1D on chip droplet counting system (which is also used in the droplet digital PCR system) and other particle counting systems including flow cytometry suffer from low throughput: they typically operate at 1,000s particles per s and are only able to analyse a total of 100,000s to 1 million droplets (or a total sample volume of tens of microlitre)31,34. T
OUAA football preview

While the big six the defending champion Western Mustangs, Ottawa Gee Gees, Laurier Golden Hawks, McMaster Marauders, Guelph Gryphons and Queen Golden Gaels have held all the playoff spots the past four years, each team has significant holes to fill.

The runnerup Gee Gees don have national MVP/quarterback Brad Sinopoli (Calgary Stampeders), the Gaels lose mammoth left tackle Matt O (Cincinnati Bengals camp) and the Golden Hawks are without defensive player of the year Giancarlo Rapanaro.

do feel like there is a bit of a changing of the guard, McMaster copy h hermes bracelet coach Stefan Ptaszek said. of us have had some graduates and seniors moving on. The team that answers the questions most decisively is likely going to be on top in the OUA. to break playoff droughts are the Windsor Lancers, the University of Toronto Varsity Blues, the York Lions and the Waterloo Warriors, who return to the league after a school imposed, one year suspension in the wake of a steroid scandal.

had 60 kids with us last year for scrimmages and practices, Warriors assistant coach Marshall Bingeman said. reality, it been the longest training camp in CIS history. Lancers have replaced 13 year bench boss Mike Morencie with rookie head man Joe D while the Blues have a Hamilton flavour with ex Ticat players Greg Gary and Donnavan Carter as new head coach and defensive co ordinator, respectively.

like McMaster a lot, I like Western, Gary said. that, we all kind of bunched up. Some of the teams like ourselves are trying to figure out how to be competitive. winner of the 104th Yates Cup game on Nov. 12 will have a decent shot of making it to the Vanier Cup on Nov. 25 in Vancouver, which is back as part of the Grey Cup week festivities for the first time since 2007 in Toronto.

The OUA winner travels to Moncton to face the hermes enamel bracelet replica Atlantic champ in the Uteck Bowl a national semifinal on Nov. 19.

Just one Atlantic team the Saint Mary Huskies in 2007 has advanced to the Vanier Cup in the past seven years.


Last season: 7 1 (Beat Ottawa in Yates Cup, lost to Laval in CIS semifinal)

Coach: Greg Marshall (24 8 in four seasons at Western)

Returning starters: 15 (nine offence, six defence)

Who to watch: DT Mike Van Praet (OUA all star), SB Zach Bull, QB Donnie Marshall

Hello: RB Tyler Varga (turned down offers from Baylor and Dartmouth), OL Winston Miles (member of Team World)

Farewell: LB John Surla (CFL Montreal), DB Craig Butler (CFL Saskatchewan), WR Nick Trevail

The skinny: Defensive co ordinator Paul Gleason has to replace all of last year starting linebackers . Deep at running hermes clic clac fake back with talented freshman Varga, Nathan Riva and Jerimy Hipperson . Have been in Yates Cup game past four years, winning three . Ranked No. 2 in first national poll.


Last season: 6 2 (lost to Western in OUA semifinal)

Coach: Stefan Ptaszek (27 13 in five seasons)

Returning starters: 16 (six offence, 10 defence)

Who to watch: QB Kyle Quinlan, WR Michael DiCroce, LB Ryan Chmielewski (all OUA all stars)

Hello: WR Max Cameron (Team Ontario), DB Nick Shortill (York Region high school MVP)

Farewell: OL Justin Glover (Two time all Canadian), OL hermes bracelet replica cheap Michael Hoy (two time OUA all star)

The skinny: Star kicker Tyler Crapigna ready to go after breaking leg last year . Quinlan spent time in CFL training camp with Hamilton as part of league internship program for young QBs . DiCroce one of top all around threats in league . Three starting OL are gone . Team, ranked third in nation, looking to make first Yates Cup appearance since 2004.
Objetivos de Desarrollo del Milenio de la ONU

Indicadores de los Objetivos de Desarrollo del

El sitio web de los Indicadores de Objetivos de Desarrollo del presenta los datos oficiales, definiciones, metodologas y fuentes para ms de 60 indicadores que sirven para medir el progreso alcanzado en la consecucin de los Objetivos de Desarrollo del . Los datos y anlisis son productos del trabajo del Grupo Interinstitucional y de Expertos sobre los Indicadores de los Objetivos de Desarrollo del (IAEG, en ingls), coordinado por la Divisin de Estadstica de las Naciones Unidas. Adems en el sitio web tambin estn disponibles los replica hermes clic h bracelet documentos e informes oficiales elaborados por el Grupo. Los enlaces a documentos y a sitios relacionados, as como las noticias actualizadas constantemente, le mantendrn al da acerca de las actividades en curso relacionadas con los Objetivos de Desarrollo del .

Mecanismo de supervisin de los objetivos de desarrollo delEl Mecanismo de supervisin de los Objetivos de Desarrollo del muestra el progreso de cada pas en sus esfuerzos por alcanzar los objetivos. Con la fecha lmite en 2015 cada vez ms cerca, ahora es ms importante que nunca saber qu objetivos van por buen camino y cules necesitan mayores esfuerzos o apoyo, tanto a escala nacional como internacional. El Mecanismo de supervisin de los Objetivos de Desarrollo del est concebido como instrumento para que los encargados de la formulacin de polticas, los especialistas en desarrollo, los periodistas, los estudiantes y otras personas, realicen un seguimiento del progreso mediante mapas interactivos y perfiles especficos de cada pas, conozcan los desafos y logros de cada pas, reciban las ltimas noticias y presten apoyo a las organizaciones que trabajan para alcanzar los Objetivos de Desarrollo del en todo el mundo.

Portal de datos de la ONU

El portal de datos de la ONU, UNdata, desarrollado por la Divisin de Estadstica del Departamento de Asuntos Econmicos hermes h bracelet fake y Sociales de las Naciones Unidas, pone al alcance de todos las bases de datos estadsticas de la ONU a travs de un nico punto de entrada en Internet por el cual los usuarios pueden realizar bsquedas y descargar fuentes estadsticas del sistema de las Naciones Unidas. El diseo innovador permite a los usuarios acceder a numerosas bases de datos de la ONU, ya sea buscando series de h hermes bracelet fake datos o mediante bsquedas por palabra clave. Actualmente, existen 14 bases de datos y 6 glosarios que contienen ms de 55 millones de datos y cubren un amplio rango de estadsticas que incluyen poblacin, industria, energa, comercio y cuentas nacionales. es un robusto sistema de base de datos que se encarga de supervisar el progreso alcanzado en la consecucin de los Objetivos de Desarrollo del . Genera tablas, grficos y mapas para informes y presentaciones. ha sido desarrollado en cooperacin con el sistema de las Naciones Unidas y mediante la adaptaci de la tecnologa del replica hermes bracelet enamel banco de datos ChildInfo del UNICEF. La base de datos organiza los indicadores por perodos de tiempo y reas geogrficas para hacer un seguimiento de los compromisos con el desarrollo humano sostenible.

Seguimiento y estadsticas de UNICEF

UNICEF mide la situacin de los nios y las mujeres y realiza un seguimiento del progreso mediante la recopilacin y el anlisis de datos. Mantiene y actualiza bases de datos internacionales y promueve la difusin de datos basados en pruebas para la planificacin y la promocin. UNICEF es el organismo lder de las Naciones Unidas responsable del control mundial de los Objetivos de Desarrollo del relacionados con los nios.
Palisades Boulder Field Trail

This awesome hike begins from the highest point of the New Jersey Palisades and steeply descends 530 feet to replica hermes h bracelets the Hudson River. Part of the trail is an obstacle course through a sea of boulders and part of it is a series of stone steps leading downward. Wear sturdy hiking shoes because some of the huge rocks are smooth and slippery particularly when weather conditions are wet or icy. Although this hike is strenuous in places, it's well worth any muscle aches you may have afterward, for the views beneath volcanically formed vertical columns are exceptional. No doubt, Italian explorer Giovanni de Verrazano was impressed when he discovered the Palisades in 1524, which he referred to as a "fence of stakes." Heavy quarrying once threatened this area, but when Congress established the Palisades Interstate Park Commission, the natural beauty of these basaltic cliffs was preserved for all to enjoy. The cliffs of the New Jersey section, whose present appearance dates back about 12,000 years to the Ice Age, have been named a National Historic Landmark and also a National Natural Landmark.

This awesome hike begins from the highest point of the New Jersey Palisades and steeply descends 530 feet to the Hudson River. Part of the trail is an obstacle course through a sea of boulders and part of it is a series of stone steps leading downward. Wear sturdy hiking shoes because some of the huge rocks are smooth and slippery particularly when weather conditions are wet or icy. Although this hike is strenuous in places, it's well worth any muscle aches you may have afterward, for the views beneath volcanically formed vertical columns are exceptional. No doubt, Italian explorer Giovanni de Verrazano was impressed when he discovered the Palisades in 1524, which he referred hermes clic h bracelet fake to as a "fence of knock off hermes bracelet stakes." Heavy quarrying once threatened replica hermes enamel bracelet this area, but when Congress established the Palisades Interstate Park Commission, the natural beauty of these basaltic cliffs was preserved for all to enjoy. The cliffs of the New Jersey section, whose present appearance dates back about 12,000 years to the Ice Age, have been named a National Historic Landmark and also a National Natural Landmark.

This hike is awesome! I started early at 7 am. Started at the interstate area, headed down the stone steps to the base of the waterfall, then headed south along the river across the boulder field. The sun was smiling, and the hawks were flying the whole time. A few hikers passed by, but by no means a festival. There are several options to complete the loop. I chose to stay on the river trail up to the park headquarters, affording a much longer loop. It was well worth the effort. Will post pics shortly.
old woman dead after Hwy 8 crash hermes Constance bag blue replica in Stoney Creek

The child was the granddaughter of the woman's neighbour, says Det. Const. Wes Wilson of the Hamilton Police Collision Reconstruction Unit (CRU). The girl's buy Hermes bags replica grandmother wasn't feeling well, so the woman offered to escort her across Highway 8, just east of hermes Birkin Bag replica Green Road in Stoney Creek, so the girl could attend St. Francis Xavier Catholic Church for a confirmation ceremony. Friday. The girl was taken to hospital, where she was treated for minor injuries and released.

Wilson says there was imitation hermes Birkin bag price no "criminality" involved, meaning the 42 year old woman driving the van was not impaired by drugs or alcohol and was not speeding. The van will undergo a mechanical inspection Monday.

No charges have been laid, says Wilson, and the investigation is ongoing. Any witnesses are asked to call him at 905 546 4755.

The westbound lanes of Highway 8 from King Street East to Green Road were closed until midnight while the CRU investigated.

It was the second fatality involving a pedestrian this week. The CRU is now identifying Richard Thelland, as the 48 year old man from Val Caron, near Sudbury, as the man killed when struck by a tow truck at a Stoney Creek gas station Monday.
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