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"The Alexander Technique is a method for improving motor performance by integrating the voluntary and reflex components of a movement in such a way that the voluntary does not interfere with the reflex and the reflex facilitates the voluntary."
Frank Pierce Jones
The Alexander Technique is not a running method; it is a method that addresses the posture that supports running, and, indeed, all movement, and is, therefore, an excellent technique for improving running form and avoiding injury. What I have written in these pages on running, gleaned from 40 years of running and some 35 years in the Alexander Technique, are observations on how runners with excellent posture can run. It is not intended as a "how-to", but more as a "what-not-to-do."
I won't attempt here to explain the Alexander Technique, as there are plenty of good books that go into great depth, including Alexander's own. And, at any rate, there is no replacement for private lessons with a certified Alexander Technique teacher (one certified by STAT, or one of its affiliates, such as AmSAT in the U.S. or CanSTAT in Canada – one should be wary of teachers who don't have STAT affiliate certification). I will, however, briefly explain how I understand the physiology behind the Alexander Technique and how it applies to running.
In vertebrates, movement is initiated, organized, and distributed through the spine. (see The Spinal Engine, which explains how the movement of the legs and arms in running originates with spinal movement) In fact, it is possible to measure muscle action in the upper neck preparatory to the movement of a finger (The Head-Neck Sensory Motor System, ed. Alain Berthoz). The first part of any movement, including walking, running and simply breathing [see: Breathing and running ] involves the adjustment of the head's weight, because the movement of a limb implies a change in the balance of the whole body. Further, movement of a limb demands the support of the spine. The spinal musculature is composed of chains of smaller muscles, which, when co-ordinated, can powerfully extend the spine against the contraction of the large muscles which attach the limbs to the spine. For example, when one lifts a heavy object with an arm, that action should not compress or bend the spine – when a leg extends or recovers in running, that movement should not result in arching of the lower back, or tilting of the pelvis – the extensor muscles of the spine reflexively act to resist the contraction of the large muscles of the limbs, and, in fact, actually prepare to do so before the movement of an arm or leg even begins. At least, this is what happens when good posture is present. In actuality, what we see in many people is quite different: their necks and backs shorten and contract with the effort of moving their limbs. Runners hunch and arch, effectively compressing their trunks and retracting their limbs. They have learned to work against themselves.
At birth, we can see reflexes that will control posture and movement already at work. A normal child has the balanced and interactive muscle tone that defines good posture. Good posture is movement, not positioning. Certain extensor reflexes can be immediately tested in newborns – pressure against the ball of a foot will elicit counter pressure, demonstrating how the body has evolved to lengthen against the force of gravity (as occurs in natural running), or, indeed against any exterior force placed upon it. One can also test, in infants, reflexes that protectively retract the limbs, and completely inhibit the extensor reflexes of the spine. The Moro reflex is a reflex, studied in apes and in man, which can best be described as a grasping reflex. Take an infant and make a sudden movement as if to drop it (well, don't actually do this, take the word of researchers who've studied it), and you will see a stereotyped two-phase response. The first is extensor – reaching out with the hands and arms, the second, flexor, retracting, and adducting the arms and legs and collapsing the neck and spine. In arboreal apes, the purpose of this reflex is evident – the infant must grasp its parent in a time of flight, as the parent needs both limbs available. In most environments, this reflex is only occasionally stimulated. In modern man, it is constantly stimulated.
To be precise, the Moro reflex does not persist beyond a few months of age. However, similar responses do persist and are referred to as startle responses. They look very much the same as the Moro reflex, demonstrating a strong and rapid grasping response, during which the extensor reflexes are inhibited. These retractive, grasping reflexes are stimulated by fear and pain. You can see them at work when you surprise someone. You can see them at work when someone is injured. And you can see them at work when someone is afraid, or simply worried. Children who begin to hunch over at their desks at school, grasping their pencils as if they were life preservers, are demonstrating the stimulation of grasping responses. As I mentioned, this is not a problem if the stimuli are rare and of brief duration, but in most early learning environments they are frequent and sustained. Children are afraid of not doing well, of not being liked, of not being understood. So they retract and hunch over. If they succeed in their tasks in spite of their retracted state, they learn to retract as a means of self-control. Eventually, the extensor reflexes of their spine are constantly inhibited, and all movement becomes more effortful. They over-stabilize their joints and move against the resistance of contracted muscles. Muscle work is no longer reflexively distributed through the spinal musculature, and is instead focused on individual muscles. And then we see what we identify as poor posture – hunched or arched backs, rounded shoulders, shortening necks, etc. The obvious and incorrect response to poorly positioned body parts is to "properly" position them. So we attempt, or we tell children to attempt to stand straight, to pull their shoulders back and down, and we use the incorrect notion of alignment to support our wrong conception of good posture. (see The Myth of Alignment) So much poor advice that we read about running form is about how to "hold" the body. Runners are told to keep the shoulders down, to cup the hands, to keep the elbows at 90 degrees, to recover the legs with the hamstrings, to shorten their strides, to keep the body aligned, and there are running coaches who insist that one can improve running posture by strengthening muscles (you cannot). The problem is, if a shoulder is rounded forward due to the installed pattern of constant retraction, pulling the shoulder back will simply add new effort and increase fixity in the shoulder joint. Trying to stand up straight will simply be adding more work to an already over-worked musculature.
This is where the Alexander Technique comes in. With gentle hands-on work, an Alexander Technique teacher brings a student's awareness to his stabilized articulations and guides the student to permit freedom and movement. The repeated experience of hands-on guidance increases the student's awareness of himself and teaches him to remove retraction and fixity from neutral posture. He may then learn to initiate movement without preparatory hardening and retraction. Hands-on private lessons are essential, because habitual preparation is very difficult to inhibit, and movement initiation by a teacher removes responsibility for the beginning of movement, making it far easier for the student to allow himself to move without preparation. Actually, good posture is dynamic, a condition in which all muscles are lightly active, from which all movement may be accomplished without any preparatory hardening or stabilizing. Running begins, in all vertebrates, with a release of the head and lengthening of the spine.
Again, many of the muscles which extend the spine against the action of large muscles of the limbs are of small mass and are only strong when organized in chains. The constant habitual retraction and adduction of the limbs inhibit spinal extensors, so it is important that one learn to allow the head and limbs to release outward in order to re-establish the spinal extensors. One cannot simply "straighten-up" to good effect.
So, in no particular order, here are a series of articles on aspects of running form. Hopefully, they can serve as guidelines for what happens in running when good use of the body is optimal. My unique contribution to running is my theory of the biarticular muscles in running (see: The Biarticular Muscles in Running).
Good running! Find a certified teacher of the Alexander Technique and take a few private lessons. Here is where to find a teacher:
The Society of Teachers of the Alexander Technique (England and international members):
The American Society of Teachers of the Alexander Technique (U.S.)
The Canadian Society of Teachers of the Alexander Technique


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401-K review and rollover can be a rewarding process
Published 8:35 pm Friday, February 21, 2014
Your 401(k) offers tax-deductible contributions, tax-deferred growth of earnings potential and a variety of investment options — so it's a great tool for building retirement savings. Yet like all tools, your 401(k) must be used properly to get the best results. That's why you should review your 401(k) at least annually and make whatever adjustments are needed.
Depending on where you work, you may get some 401(k) review help from your plan provider. But if that assistance isn't available, you might want to consult with a financial professional to make sure you're getting the maximum benefit from your plan.
As you begin to review your 401(k), your first question should probably be this: "How much should I contribute?" At the very least, try to put in enough to receive your employer's matching contribution, if one is offered. If you don't earn this match, you are essentially walking away from "free money." Beyond this, though, the amount you put into your 401(k) might depend on what other retirement savings vehicles you have available. For instance, if you're eligible, you may also want to contribute to a Roth IRA, which offers tax-free growth potential, provided you've had your account for five years and don't start taking withdrawals until you're 59-and-a-half.
Of course, it's not only how much you put into your 401(k) that determines its success — it's also how you choose to allocate your investment dollars. (Keep in mind that asset allocation does not guarantee a profit or protect against loss.) Your 401(k) may have a dozen or more investment choices, such as stock funds, bond funds and money market funds. To choose the right investment mix, you'll need to consider a variety of factors, including these:
u Your age — Generally speaking, the younger you are, the more aggressive you can afford to be with your 401(k) investments, because you'll have decades in which to potentially overcome the inevitable down periods of the market. As you get older, you may wish to invest somewhat more conservatively, but you'll still need some growth potential in your 401(k) portfolio.
u Your goals — Everyone has different goals for retirement. You might want to retire early and travel the world, while your co-worker desires to work as long as possible and then, upon retirement, stay close to home and pursue hobbies. Because you each have different goals, with different income needs, you also may need to follow different investment strategies within your 401(k).
u Your other retirement income sources — If you have a variety of retirement income sources — a pension from another job, an IRA, a spouse with generous retirement benefits — you may need to invest differently, perhaps less aggressively, than if you had fewer options for retirement income.
Apart from putting away as much as you can into your 401(k) and choosing the right investment mix, what else can you do to get the most out of your plan? Here's a suggestion: ff you have worked at various jobs and acquired multiple 401(k)s, consider rolling them over into one account. You might save money on fees and reduce paperwork, but more importantly, you'll be able to concentrate your resources and pursue a unified investment approach, with your investment dollars working together toward your ultimate retirement goals.
As you can see, a 401(k) review and rollover can reward you in many ways — so do whatever it takes to maximize your 401(k)'s performance.
This article was written by Edward Jones for use by your local Edward Jones Financial Advisor.

subset: commoncrawl


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Barbara Frietchie
This poem was written in strict conformity to the account of the incident as I had it from respectable and trustworthy sources. It has since been the subject of a good deal of conflicting testimony, and the story was probably incorrect in some of its details. It is admitted by all that Barbara Frietchie was no myth, but a worthy and highly esteemed gentlewoman, intensely loyal and a hater of the Slavery Rebellion, holding her Union flag sacred and keeping it with her Bible; that when the Confederates halted before her house, and entered her dooryard, she denounced them in vigorous language, shook her cane in their faces, and drove them out; and when General Burnside's troops followed close upon Jackson's, she waved her flag and cheered them. It is stated that May Qnantrell, a brave and loyal lady in another part of the city, did wave her flag in sight of the Confederates. It is possible that there has been a blending of the two incidents.
Up from the meadows rich with corn,
Clear in the cool September morn.
The clustered spires of Frederick stand
Green-walled by the hills of Maryland.
Round about them orchards sweep,
Apple and peach tree fruited deep,
Fair as the garden of the Lord
To the eyes of the famished rebel horde,
On that pleasant morn of the early fall
When Lee marched over the mountain-wall;
Over the mountains winding down,
Horse and foot, into Frederick town.
Forty flags with their silver stars,
Forty flags with their crimson bars,
Flapped in the morning wind: the sun
Of noon looked down, and saw not one.
Up rose old Barbara Frietchie then,
Bowed with her fourscore years and ten;
Bravest of all in Frederick town,
She took up the flag the men hauled down;
In her attic window the staff she set,
To show that one heart was loyal yet.
Up the street came the rebel tread,
Stonewall Jackson riding ahead.
Under his slouched hat left and right
He glanced; the old flag met his sight.
"Halt!"–the dust-brown ranks stood fast.
"Fire!"–out blazed the rifle-blast.
It shivered the window, pane and sash;
It rent the banner with seam and gash.
Quick, as it fell, from the broken staff
Dame Barbara snatched the silken scarf.
She leaned far out on the window-sill,
And shook it forth with a royal will.
"Shoot, if you must, this old gray head,
But spare your country's flag," she said.
A shade of sadness, a blush of shame,
Over the face of the leader came;
The nobler nature within him stirred
To life at that woman's deed and word.
"Who touches a hair of yon gray head
Dies like a dog! March on!" he said.
All day long through Frederick street
Sounded the tread of marching feet.
All day long that free flag tost
Over the heads of the rebel host.
Ever its torn folds rose and fell
On the loyal winds that loved it well;
And through the hill-gaps sunset light
Shone over it with a warm good-night.
Barbara Frietchie's work is o'er,
And the Rebel rides on his raids no more.
Honor to her! and let a tear
Fall, for her sake, on Stonewall's bier.
Over Barbara Frietchie's grave,
Flag of Freedom and Union, wave!
Peace and order and beauty draw
Round thy symbol of light and law;
And ever the stars above look down
On thy stars below in Frederick town!


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How to Make Meaning of the Holidays as a Non-Christian
The recent media attention to whether Jesus was white, whether Santa is white, and the so-called "war on Christmas" distract from a question that has plagued me my whole life: What's a non-Christian South Asian American to do during Christmas?
Growing up, my Indian immigrant family had a variety of responses to the holiday season. My mom, who attended Catholic schools in India as a child, was particularly fond of the holiday. She always wanted a tree and took us to take photos with Santa at the mall. One year, she even had our Catholic babysitter take us to midnight Mass. That night, my brother and I followed Sandra's children up to the priest to get communion — and were angrily yelled at by a parishioner for not knowing what to do after receiving it. We never returned to that service.
Another year, after my grandfather died, we were extending the usual Hindu tradition of no celebrations for 13 days after a death, but my mom still insisted on making a tree shape out of lights to hang in our window. It seemed as if, rather than the repetition of traditions, our holiday season was marked by the search for them.
There were always gifts (on Diwali and, soon after, on Christmas) and time together as my parents, who both worked full time, were around more. We often had visits from family who were scattered around the U.S. or other Indian immigrant families taking advantage of the time away from work at liquor stores, driving taxis, or at hospitals and tech companies. Since college, my holiday traditions became related to overseas travel over the long break as a student and then as a professor.
As a new mother this year, I have started my own quest for tradition. My partner and I are raising a child who has Hindu, Sikh, and Christian heritage. We want to pass down some aspects of our experiences as South Asian Americans.
My husband grew up in a Syrian Christian community of south India, a community that traces its lineage to Saint Thomas and the many versions of Christianity that have developed in the country since the 1st century AD.
However, my partner has not been a practicing Christian for over two decades. We recently bought a tree. I love the smell and the somewhat-hidden history of ancient Egyptians displaying evergreen boughs around the winter solstice as a symbol of renewal, regeneration, and life over death. (Reportedly, the tradition of decorated trees in homes came to the United States via German settlers in the 1800s.)
We've decided to call our tree a "family tree" and, instead of lights and store-bought ornaments, we put photos of relatives, loved ones, ancestors, and small items that represent our families on the branches. It reminds me a bit of the Dia de Los Muertos altars I saw Chicano friends putting together when I was growing up in California. But we have plenty of "living" and other images — my newborn nephew's photo, an ornament made in grade school by my brother, and a set of handmade paper mâché stars bought on a family trip to Goa when I was eight years old.
Who knows if the tradition will stick. We may keep searching with no firm outcome as my family did. I imagine this is what many immigrant families to the U.S. have done. In fact, belief in Santa Claus and his reindeers, the displaying of trees, and the drinking of egg nog were all brought by immigrants from different parts and "indigenized" here in the U.S. over time.
Regardless of where our family traditions end up, for now, looking at pictures of loved ones past and present, with the smell of pine, brings us smiles and feelings of gratitude as we begin a new year full of promise.


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Harvest Fresh Cuts Expansion
Client: | Harvest Fresh Cuts |
Location: | Wacol, Brisbane |
Scope overview: | Following the successful design and construction of the 2,800m2 facility at Wacol, Wiley were engaged again to design and construct the expansion. |
Project Description
Harvest Fresh Cuts are a subsidiary company of the parent One Harvest.
With facilities in Brisbane, Perth and Bairnsdale they specialise in producing a variety of salad bags, kits and bowls, value-added vegetables, prepared deli salads and fresh chilled meals.
Wiley began their relationship with Harvest Fresh Cuts over a decade ago when they were contracted to design and construct a 2,800m2 facility at Wacol, Brisbane. This facility was recognised at the time as one of the most advanced food processing plants in the world.
Wiley was once again approached by Harvest to design and construct an expansion of this facility.
The expansion would incorporate:
- Addition of a new production line
- Relocation of two existing production lines to allow for raw materials and finished goods storage.
Wiley worked with Harvest during initial meetings with overseas equipment suppliers providing valuable insight into the coordination of the process design, management and implementation of this equipment.
Harvest contracted Wiley to continue with the construction phase, and assist through the complete design and commissioning of the equipment.
- Working with a foreign equipment supplier to meet Australian Standards for electrical, safety guarding and general access
- Extremely short shelf life of product. This limited extensive shut downs in which works could be completed
- Managing construction and equipment installation without impinging on the current operations of the facility
- Maintaining client production rates while onsite work took place.
- Provide information on Australian equipment standards and installation procedures to overseas suppliers
- Advise on project management procedures and timelines to suit limited shut down periods
- Provide guidance on services upgrades and maintenance of production rates.
- Determine adequate space for the new production line and relocation of existing lines
- Layout designed for maximum throughput
- Design for compliance with health and hygiene requirements
- Design for integration of new and existing processes within the completely refrigerated processing space, temperature controlled at around 4°C with a humidity range from low to high.
- Coordination of services and assurance that imported, specialised processing equipment was compatible with Australian requirements and conditions
- Work with client to ensure equipment complied with client's requirements and expectations.
- Coordination of construction activities with process equipment installation and commissioning
- Working around the clock to achieve the required results with minimum shutdown periods – including moving an entire wall in 48 hours
- Staging to meet timeframes of equipment coming from overseas
- Minimising impact on existing services and coordinating contractors
- Upgrade council services concurrent to work on-site – external to site then into the site.
Wiley's specialist knowledge in process design and management, aligned with a holistic approach to master planning and project management, has given the client the potential for continuous future expansion—a facility that will grow with the business.
This ensures Harvest Freshcuts will continue to meet demand for minimally processed fresh, healthy and conveniently packed food products.


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Grand Gourmet Flagship Store
To show respect to these handmade products, the shop design was generated by a custom made computer algorithm, the fractal hexagon pattern gives the shop strong identity and organized all functions. The making process involved many cutting edge technologies and traditional handcrafts. The creative combination of the traditional handcraft and the modern technology in design and construction process makes the shop unique.
Continue reading


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CSI: Hard Evidence
Fans of the CSI series have a 4th chance to become part of the popular crime-busting team…but will the evidence reveal a satisfying game? The series that made tedious lab work look cool actually translates well to the adventure world. Of course, I speak as a fan of the original series, but I think the average gamer might find something to enjoy in CSI: Hard Evidence. It's not the challenge of the season, but it IS absorbing.
According to the official web site, CSI: Hard Evidence offers an even more authentic experience than ever before, with improved visuals, more lab equipment to use, and more investigative features. I didn't playCSI: Dimensions of Murder, but compared to CSI: Dark Motives, I tend to agree. The 5 episodes lasted longer, were more in-depth, and had more unforeseeable plot twists.
I was also more impressed by the well-written dialogue in this game. Snappy puns were sprinkled liberally throughout, and the voice actors delivered their lines in a realistic manner. Now, they obviously aren't as comfortable reading lines as they are with acting, but no one's performance was flat enough to stand out.
As always, you'll play the new recruit, investigating everything from a cabbie burned to a crisp to a Wild West shootout done Vegas style. Along the way, you'll have the chance to get in good with Gil (and open bonus material) by collecting those creepy-crawly bugs he loves so much. As I said, each of the stories is engrossing; they're just not that difficult.
Now, if you're in the right mood to play what basically amounts to an interactive, high-tech detective story, Hard Evidence is a beautiful thing. My fear is that unless they are fans, seasoned adventure gamers my feel a distinct lack of challenge. This isn't because the stories are facile, but because the game is designed to lead you to each next step.
For example, you have a wide array of collection and detection tools to choose from. However, the game will only allow you to choose between two or three at a time, meaning that you don't have to learn the proper use for each tool. Team members are sometimes too quick to tell you the next step, even when you don't ask for a hint. You're even told when an area doesn't have anything useful by being given a bonus point for thoroughness. Another easy-making aspect: each area you've searched or item you've collected will be tagged gray or green. If it's green, you've done all you can do with it, so you know that whatever you're missing isn't there. With all this help plus the 29 (I believe) hints you're allowed per episode, there are few opportunities to be truly stuck.
The actual game play will be familiar to any adventure gamer. Green arrows indicate hot spots, and you click to go there. Before collecting evidence for inventory, you'll have to use the proper collection (and sometimes detection) tools. Once you choose the proper tool, the icon will change to a grabby hand. You can further investigate inventory items, which—like everything else in the game—are accessible through your PDA.
But just as in the television show, collecting evidence is only part of the process. The real work takes place in the lab, where you'll have access to a comparison microscope, a trace analysis computer, DNA and chemical analyzers and the assembly table. If you try to confirm a DNA, fingerprint, or chemical match that isn't correct, you'll be scolded. It won't, however, count against your evaluation as I originally assumed it would.
Now, I've mentioned the evaluation a couple of times, so I should explain it. Essentially, you're judged by whether you found and processed all the evidence in each episode, and how many (if any) hints you needed to finish. Bonus points (which offer access to bonus materials) are gained by collecting bugs and by earning "thoroughness points," i.e., clicking on every possible hot spot. Unless you're really a perfectionist who has to finish with a Master Investigator grade, this won't mean much. After all, the fact that you've reached the review stage indicates that you've completed the episode. As for the bonus materials, they consist of sketches, story boards, etc.—things that some people may enjoy.
The graphics in CSI: Hard Evidence had both good and mediocre elements to them. Certain team members (Nick in particular), had a very odd look about them. I had difficulty pinpointing what the matter was, but I think it had something to do with the way their mouths moved. They seemed to "implode inward" on the face, if you can figure out what the heck I mean by that. Gil Grissom looked disturbingly like Hank Hill from King of the Hill, and Catherine Willows resembled any other top-heavy CGI babe. I thought Warrick, Greg and Captain Brass–although not perfect–were much better done.
The backgrounds were crisp and clean, most items looked realistic, and movement was fluid. Once you're in a particular screen, you can move your cursor to the edges to rotate within the scene. This gives a sort of "floating camera" effect that's easy to navigate once you get used to it. Right clicking your mouse allows you to back out of any scene until you reach the original starting point.
Because of the 3D graphics, this game may be more demanding for some computers than for the players, so be sure to check the requirements before you buy. I didn't have any particular problems, but I did notice some very slow loading times. I suspect that someone using an older computer might have more issues, although I have no evidence to support my theory.
I do have evidence of something that really irritated me: the blatant, unabashed product promotions, particularly of Visa. If it were simply a matter of the logo prominently displayed in appropriate places (the bar at the high rollers lounge; the display glass that has to be reconstructed; the cabbie's sign board), it wouldn't have been a big deal. But they went so far as to weave the fraud protection program into a story line, having Brass say at one point, "I called Visa… good folks there." There were other products being pimped as well, but they were smooth enough not to raise my ire. I could understand it if this game were being given away or sold at greatly reduced cost as a promotional piece. But this is a full-priced game, and to have advertisements foisted upon a paying public seems shady.
In the end, we all play games because of the entertainment factor. Whether or not you'll like this game depends on how important difficulty and challenge are to your enjoyment. I generally want more challenge out of my games, but I made an exception for this one because it did the job—it entertained me for several consecutive evenings. As Gil says during each evaluation, "At the end of the day, that's all that really matters." I give this game a B.
Final Grade: B |
System Requirements:
• Windows 2000 / XP / Vista
• 2.0 GHz processor or better
• 64MB 3D-accelerated video card
• 1.5GB hard drive space
• DVD-ROM drive


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Use Machine Learning Algorithms to Explore the Potential of Your High Dimensional Flow Cytometry Data Example of a 20–color Panel on CytoFLEX LX
Here you will learn how machine learning algorithms like viSNE and FlowSOM can help you identifying specific populations/subsets. We use data from a 20–color CytoFLEX LX panel and provide details on how to build a computational flow cytometry data analysis pipeline within Cytobank. We will discuss how to assess the quality of viSNE maps and FlowSOM clustering results and we will help you recognize how processing steps can affect the result quality of machine learning algorithms.
Due to the advancement of technology in the cytometry field, we are now able to measure many parameters simultaneously on a single cell level. The dimensionality of datasets has increased from traditional 4–5 color low parameter to 10–20 or more color with several commercially available instruments. The sample size per experiment is also getting increasingly larger in terms of the number of events and number of samples collected. This imposes a challenge in regards to data management, collaboration, and data visualization and analysis. Many machine learning algorithmic tools are developed for dimensionality reduction and clustering to handle this increase in data complexity (Figure 1).
Cytobank is a cloud–based analysis platform with integrated analysis algorithms, as well as a structured and secure content management system for flow cytometry and other single cell data. Cytobank's clustering, dimensionality reduction, and visualization tools (SPADE, viSNE, CITRUS, FlowSOM) leverage the scalable compute and collaborative power of the cloud, allowing large analyses to be done in parallel; the cloud–based storage provides the capability to automatically archive and easily share these data securely and safely 2.
In the following we illustrate how machine learning algorithms can help analyze high dimensional data using example data from a CytoFLEX LX flow cytometer 20 color experiment.
Figure 1. Comparison of biaxial plots with machine learning analysis of a 20–color panel cytometry data. A) N by N plot view of a 20–color panel cytometry data resulting in 190 plots. B) viSNE (or t–SNE) map view of the same data that visualize the 24–parameter information on one single map. NOTE: viSNE in Cytobank uses the Barnes–Hut implementation of the t–SNE algorithm 2,5.
Building a High Dimensional Flow Cytometry Data Analysis Pipeline with Cytobank
The unsupervised nature of algorithms such as viSNE, SPADE and FlowSOM increases the chance of discovering previously undescribed phenotypes. viSNE is a dimensionality reduction and visualization algorithm that creates a two–dimensional view of high–parameter data, allowing you to identify interesting and rare biological subsets. viSNE is often used as a first step for exploratory data analysis and in Cytobank can be easily linked to clustering algorithms1. FlowSOM is a Self Organizing Map algorithm that clusters phenotypically similar cells in a simple–to–interpret minimum spanning tree for fast, intuitive analysis and presentation of large high–dimensional data sets5. Combining clustering and viSNE data and displaying clustering data on a viSNE map alongside your native data can help with clustering algorithm optimization and with the assessment of cluster identity. In the next section we provide guidance on how to perfom the required steps as outlined in Figure 2.
Figure 2. High Dimensional Flow Cytometry Data Analysis Pipeline with Cytobank. Key steps outlined for biomarker discovery analysis pipeline. 1) Data acquisition 2) Data compensation, transformation and quality control and 3) data cleanup and an optional pre–gating 4) Exploratory data analysis with viSNE 5) clustering and biomarker Discovery, 6)visualizations to communicate results.
1) Data acquisition
During the experiment setup and data acquisition process, it is important to consider the sample size required for statistical validity, as well as control samples depending on your research questions. If you run your samples in multiple batches, potential batch effects also need to be assessed and controlled.
2) Data compensation, transformation and quality control
Cytobank offers tools to prepare data for the subsequent analysis using machine learning algorithms. Alternatively, Kaluza Analysis Software and the Kaluza Cyotbank plugin may be used for fluorescence compensation and data transformation.
- Compensation: Fluorescence spillover of fluorochromes into channels other than their designated detection channel is corrected by fluorescence compensation. Compensation artifacts can negatively impact downstream analysis if not corrected. Even if samples are acquired with the compensation matrix already applied, it is advisable to check and correct the compensation for all samples.
Figure 3. Cytometry data need to be properly compensated prior to the advanced machine learning analysis. Example of inappropriate (A) and appropriate compensation (B).
- Scale transformation: In flow cytometry, one of the following display scales are typically used, linear, log10, arcsinh, or biexponential transformation. Cytobank will automatically set the scale equation for fluorescent channels to arcsinh for data produced by most modern digital cytometers. It is very important to set the scales correctly for the advanced machine learning algorithms, especially the scale arguments. The algorithms will read the full range of your scale no matter if they are off scale or not, i.e. scale min and max do not matter, but the scale arguments value will affect the results. Please refer to our blog post on how to scale cytometry data effectively.
Figure 4. Scale transformation settings. An example of bad (A) and good scales (B). The CD4 channel was scaled improperly on the left, with a dip around the zero region, where all the negative values should form a single peak around the region, as shown on the right.
- Quality control: The stability of the sample acquisition process can be evaluated by displaying scatter and fluorescence versus time data. Gaps in data acquisition may indicate disruptions of the sample flow by blockages or bubbles. This display allows the user to restrict the analysis to parts of the acquisition when sample flow and signal detection were stable.
Figure 5. Quality control. An example of stable data acquisition shown by the constant fluorescence intensity over time and lack of gaps.
3) Data cleanup and an optional pre–gating
Remove unwanted events, such as from the dump channel, debris, doublets, and dead cells, by pre–gating. These events do not add information to the downstream analysis, could negatively impact data display and confound statistical results if not identified and excluded appropriately. Depending on the desired data visualization and research questions, it may be of value to pre–gate on the population of interest for further analysis.
Figure 6. Data pre–processing by A) gate on single cells, B) gate out debris and C) gate on live cells D) gating on a major subset like CD45+ leukocytes or lymphocytes is optional.
4) Exploratory data analysis with viSNE
- How to set up a viSNE run
- Select the pre–gated population and the files for all of the samples to be analyzed. Set the total number of events to the maximum number of events for viSNE, and select equal sampling.
- Select the clustering channels to use for viSNE. You should exclude any channels used to define subsets during pre–gating, and include all channels that you want to use to define populations. Do not mix channels with linear scales and arcsinh scales in the channel selection, for example by selecting the time parameter or the scatter channels with the fluorescence channels together.
- For fluorescent data, select the appropriate compensation matrix just as you would for any other analysis.
- Adjust the number of iterations and perplexity based on the number of events. For fluorescent cytometry data, a good starting point for iterations is roughly 1000 iterations for every 100k events. NOTE: Mass cytometry data typically requires fewer iterations than fluorescent data. Perplexity can be thought of as a rough guess for the number of close neighbors (or similar points) any given event or observation will have, therefore it is specific to a given data set. Increasing perplexity can also have a big impact on algorithm run time. We recommend starting with a perplexity of 30 if the number of events in the viSNE is less than 1M, 50 for 1M to 1.5M events, and 70 for more than 1.5M events.
- Run the viSNE. If you need to, set up multiple viSNEs runs in parallel on the cloud.
- How to interpret the viSNE results
- Assess the viSNE map quality
Before you can use viSNE for any exploratory data analysis, you'll want to check that you don't need to fine tune any of the advanced viSNE settings. Set up a working illustration that shows the viSNE map for every file on the rows colored by every channel on the columns. Assess the viSNE map quality by examining this working illustration to make sure that it has converged well.
- Assess the viSNE map quality
Figure 7. viSNE Visualization of 20–color flow cytometry data. viSNE ran on gated live single cells, 19 markers selected as viSNE channels. viSNE setting for 100,000 events: 1000 iteration, 30 perplexity and 0.5 theta (default settings). The intensity of each marker expression, which is labeled in the top left corner of each plot, is indicated by the plot's scale on the right.
In a nicely converged viSNE map, cells that have similar expression of clustering markers will form either separate viSNE islands (if they are very different from other cell types) or separate regions within viSNE islands. A poorly converged viSNE map will have overlapping and poorly formed islands that don't separate the expression of a single marker into distinct locations on the map (Figure 8A). Cells expressing a given marker may appear in a string–like or spindly pattern. If the viSNE map is poorly converged, you should rerun viSNE with additional iterations (Figure 8B). viSNE maps can also be converged with the islands not very well separated. In this case, you should rerun viSNE with higher perplexity (Figure 8C). Rerun viSNE if needed, adjusting the settings based on your findings.
Figure 8. viSNE Visualization of 20–color flow cytometry data. viSNE ran on gated live single cells, 19 markers selected as viSNE channels. viSNE settings were chosen as indicated. The intensity of each marker expression, which is labeled on the top of each plot, is indicated by the plot's scale on the right.
Cancellations and failures for viSNE happen in only ~1% of all viSNE runs on the Cytobank platform. If you experience problems with cancellations and failures, it may be because your particular data set demands a lot of memory.
Some things you can try if you experience this are:
- rerun the original viSNE with the same settings
- rerun the viSNE with a lower perplexity and/or lower iterations
- select a subset of events before starting by gating on the time channel and then using the resulting population to split files by population if the number of selected events is much greater than the number of events sampled (visible in the setup page)
- Exploring your data with viSNE
There are several visualizations you can create with viSNE to help explore and explain your results.
- Color the viSNE map by expression of clustering markers. Create a working illustration of dot plots showing the tSNE axes colored by channel with all of the clustering channels on the columns. (see Fig. 7 above)
- If you're looking at differences in functional marker expression, color the viSNE map by expression of functional markers not used for clustering. Create a working illustration of dot plots showing the tSNE axes colored by channel with the functional marker channels on the columns (Figure 9B). This can be done using a grid layout with files on the rows organized using sample tags for outcome groups.
Figure 9. viSNE Visualization of 20–color flow cytometry data. viSNE ran on gated live lymphocytes, 6 markers were selected as viSNE channels viSNE setting for 100,000 events: 1000 iteration, 30 perplexity and 0.5 theta. The intensity of each marker expression, which is labeled on the top of each plot, is indicated by the plot's scale on the right. A shows population markers for orientation purposes B shows functional markers.
- To keep track of the cell populations you are looking at, it may be helpful to manually gate the islands of the viSNE map and then display these on the working illustration. Do this by selecting 'show gates' and using the gate label 'gate name'.
Figure 10. Gate on viSNE identified population. Use the same gating tools for dot plots on the gating window to gate on the viSNE identified population by showing the viSNE plots with third channels.
- Examples of things gone wrong
- Failing to appropriately correct for fluorescence spillover and to optimize data transformation may impact the results of dimensionality reduction with viSNE.
- Inadequate data transformation:
Figure 11: A shows improper scaled data with negative populations for several of the markers displayed being split into 2 populations. Machine learning algorithms are unable to identify this artefact and as a result assume differences between the negative events. This can prevent the viSNE map from converging and may affect coloring by channel.
Figure 11. Effect of data transformation on viSNE visualization of 20–color flow cytometry data. viSNE performed as indicated in Figure 7. Representative histograms for lymphocyte markers show the data transformation. A shows suboptimal data transformation resulting in a poorly converged viSNE map and falsely high coloring for CD19. B shows the same data with optimized transformation settings.
- Inadequate fluorescence compensation may lead to the appearance of false positive or false negative events and will impact the coloring by channel. Figure 12A shows overcompensation of spillover from the CD4 signal into the CD56 channel, resulting in the appearance of extremely low fluorescence intensity of CD4+ cells in the CD56 channel. Undercompensation of the spillover of CD8 in the CD15 channel results in these cells showing in incorrect intermediate signal intensity for CD15. Figure 12B shows results after spillover correction.
Figure 12. Effect of fluorescence spillover compensation on viSNE visualization of 20–color flow cytometry data. viSNE performed as indicated in Figure 7.
5) Clustering and biomarker discovery
Clustering algorithms such as FlowSOM and SPADE can be used to automatically identify cells of similar phenotype in an unbiased manner 3,4. It can be performed on the original uploaded experiment or it can be performed on the generated viSNE experiment and viSNE identified islands.
- Run FlowSOM on the original experiment
- Set up the FlowSOM analysis on the original experiment
- Select the pre–gated population and the files for all of the samples you determined you will include in this FlowSOM run. Set the total number of events to the proper number of events, and select equal sampling. FlowSOM is a fast algorithm that can process up to 4 million total events on the premium server.
- Select the clustering channels to use for FlowSOM. You should exclude any channels used to define subsets during pre–gating, and include all channels that you want to use to define populations. Be sure to not mix channels with linear scales and arcsinh scales with the channel selection, such as select the time parameter or the scatter channels together with fluorescence channels.
- For fluorescent data, select the appropriate compensation matrix just as you would for any other analysis.
- Choose the Algorithm settings.
- SOM selection: You may choose to create a new SOM or use an existing SOM from a previous run.
- Clustering method: choose a method that will be used for metaclustering. The default Hierarchical Consensus clustering works best in most cases.
- Choose a target number of metaclusters: The number of metaclusters for a FlowSOM run determines how many metaclusters the clusters will be organized into. The correct number of metaclusters will depend on many factors and is data specific. A good starting point is to use a number that is slightly higher than the total number of major subsets to be discovered.
- SOM selection: You may choose to create a new SOM or use an existing SOM from a previous run.
- Clustering method: choose a method that will be used for metaclustering. The default Hierarchical Consensus clustering works best in most cases.
- Choose a target number of metaclusters: The number of metaclusters for a FlowSOM run determines how many metaclusters the clusters will be organized into. The correct number of metaclusters will depend on many factors and is data specific. A good starting point is to use a number that is slightly higher than the total number of major subsets to be discovered.
- To configure the PDF output review our online support article How to Configure and Run a FlowSOM Analysis here
- Run FlowSOM by selecting the green button at the top. If you need to, set up multiple FlowSOM runs in parallel on the cloud by copying the settings and starting a new run.
- Run FlowSOM on the viSNE experiment
In order to visualize the FlowSOM clusters on a viSNE plot, you need to initiate the FlowSOM run within the previously created viSNE experiment. You may access the viSNE experiment from the original experiment by going to Actions>View experiment summary. In the Linked experiments section select the viSNE experiment on which you wish to run FlowSOM and then follow the steps outlined in section "Set up the FlowSOM analysis on the original experiment".
Optimizing FlowSOM settings
After successfully setting up and running a FlowSOM analysis, you can perform exploratory or quantitative downstream analysis on your FlowSOM results. As part of the analysis process, you'll want to assess the quality of the FlowSOM analysis, which can be done by displaying FlowSOM metaclusters on a viSNE map and with heatmaps that show the marker expression of the FlowSOM metaclusters. When a FlowSOM run completes, a new experiment is created that contains the original files with new channels added for the FlowSOM cluster ID and metacluster ID. You can access this experiment either from the FlowSOM settings page for the run (click "View created experiment" on the page–level navbar), or from the Experiment Summary page of the originating experiment (click on the FlowSOM run name and you will be taken to the FlowSOM experiment).
Overlaying FlowSOM–identified metaclusters onto a viSNE map
The Working Illustration of the newly created experiment will be configured by default to set up dot plots with metaclusters overlaid on the viSNE map if you ran viSNE first, followed by FlowSOM. This visualization can help you assess the quality of the metaclustering and can inform how you may need to iterate various settings including target number of metaclusters and normalization.
You can compare overlaid clusters on a viSNE map to a contour plot of the viSNE map to gauge whether the number of clusters generated was sufficient to capture distinct, dense populations.
Figure 13. A: Contour plot of the viSNE map. B: manual gates overlaid on viSNE map C–E: FlowSOM metaclusters are overlaid on the viSNE map. Same seed was manually set for all FlowSOM runs. Clustering channels for FlowSOM are the same as the channels used for the viSNE experiment. C: 10 metaclusters; D: 22 metaclusters; E: 35 metaclusters; F: 22 metaclusters, normalized.
Generally, the colors of the overlaid clusters should correspond to the viSNE continent definitions. If the same cluster spans multiple continents, that might indicate a need to increase the target number of clusters and/or metaclusters, or a need to enable or disable normalization. In the example above, you can see how a FlowSOM target number of metaclusters of 10 does not provide enough resolution, and a target of 35 provides too much resolution (though you could further explore this to see if, in fact, it is detecting relevant sub–populations). A target number of metaclusters of 22 appears to yield good correspondence between the FlowSOM metaclustering and the viSNE continents for the data set used here.
While normalization is often a helpful and necessary approach to transforming channel data prior to analysis, for some datasets it is not (see example above), and that can be rapidly detected via the display of FlowSOM metaclusters on a viSNE map, followed up by inspection of cluster expression. If you find that enabling normalization appears to diminish the resolution of the metaclustering, you can also try increasing the target number of metaclusters. There may be cases where enabling normalization appears to lower the resolution and result in poor quality, but by increasing the target number of metaclusters, you may be able to pull out sub–populations.
When a FlowSOM run completes, it writes FlowSOM_cluster_id and FlowSOM_metacluster_id channels into the newly generated files that comprise the FlowSOM analysis experiment. If you have run viSNE on the files first, the files in the FlowSOM analysis experiment will now contain all of the original channels and data, as well as the tSNE1 and tSNE2 channels from the viSNE run, and the new FlowSOM_cluster_id and FlowSOM_metacluster_id channels. The Working Illustration will be configured by default to set up dot plots with metaclusters overlaid on the viSNE map. (You can also run FlowSOM first, and then run viSNE – the same smart Working Illustration view will be set up following the viSNE run.)
6) Visualizations to communicate results
When running FlowSOM in an experiment containing viSNE results, you can take advantage of both the clustering and dimensionality reduction algorithms. You can use FlowSOM to get quantitative clusters to identify unique subsets and or marker expression and then use viSNE for visualization.
Figure 14. Contour map of viSNE map compared toFlowSOM metaclusters overlaid on the viSNE map.
FlowSOM was performed on the viSNE experiment with 22 metaclusters. Clustering channels for FlowSOM are the same as the channels used for the viSNE experiment.
- Identify clusters using heatmaps to visualize marker expression
Now that you have optimized your clustering algorithm run, you can proceed to view the expression of markers of interest in heatmap format. On the Working Illustration page, under "Plot Controls" under the illustration tab, select "Plot Type" –– "Heatmap". Make sure to select the appropriate channels for your heatmap in the "Channels" dimension. Adjust the other dimensions as needed. In order to create a heatmap with multiple channels, use the channels figure dimension box within the Working Illustration. Make sure to set the X axis illustration setting to Panel/Channel Values. This setting defers control of the x axis to the channels figure dimension box and allows for figures with many channels. A common mistake is to not set this axis setting correctly, resulting in every square of the heatmap having the same data value.
Figure 15. Example of how to create a heatmap to compare marker expression between metaclusters. Important settings are indicated by green boxes. FlowSOM metaclusters are listed in rows and marker expression on the column. Displayed are raw median intensity values.
The scale coloring range is the range of the color bar below the heatmap.
Global (Automatic): The coloring range is the same across heatmaps.
Individual: The coloring range is calculated per heatmap.
Global (User–defined): The coloring range is user–defined and global with an inflection point. Currently it is forced to use both sides of the color scale for this configuration.
The desired statistic and equation (e.g. fold) can be chosen using the illustration controls panel on the left side of the working illustration. Heatmaps can be made with any statistic, such as event counts, percent in gate, medians, as well as ratios of these statistics compared between samples (e.g. fold change of event counts).
By default, heatmaps display statistical values and color themselves according to raw, unscaled data values. In order to view the data in the biologically more useful transformed space, the transformed ratio should be used.
When using a comparison equation, make sure to set the control appropriately. A control of table's minimum is usually appropriate to visualize an entire heatmap in scaled mode. When using a single heatmap with many channels, however, a control that normalizes each channel to itself is likely desired. Controls using column or row would be indicated for this, depending on how channels are arranged.
While it is possible to perform viSNE, SPADE and FlowSOM calculations using desktop software, the computer cannot be used for other tasks during this time and the available compute may limit the number of events that can be analyzed per sample, requiring down–sampling. Also, only one run at a time can be performed, slowing down optimization of algorithm settings. Cloud–based data analysis does not suffer from these limitations, multiple runs can be run in parallel, ie varying algorithm settings, and do not utilize desktop compute power.
Combining dimensionality reduction using viSNE, clustering with FlowSOM and visualization of expression patterns using heatmaps can be combined into a data analysis pipeline that will allow unbiased identification of cellular subsets. Performing this integrated workflow on the Cytobank platform allows users to easily combine algorithms and take advantage of smart visualization options. Running analyses in the cloud allows the user to perform different runs in parallel to optimize settings faster.
- Amir, E. D., Davis, K. L., Tadmor, M. D., Simonds, E. F., Levine, J. H., Bendall, S. C., Shenfeld, D. K., Krishnaswamy, S., Nolan, G. P., & Pe'er, D. (2013). ViSNE enables visualization of high dimensional single–cell data and reveals phenotypic heterogeneity of leukemia. Nature Biotechnology, 31(6), 545–552. https://doi.org/10.1038/nbt.2594
- Kotecha, N., Krutzik, P. O., & Irish, J. M. (2010). Web–Based Analysis and Publication of Flow Cytometry Experiments. Current Protocols in Cytometry, 53(1), 10.17.1–10.17.24. https://doi.org/10.1002/0471142956.cy1017s53
- Qiu, P., Simonds, E. F., Bendall, S. C., Gibbs, K. D., Bruggner, R. V., Linderman, M. D., Sachs, K., Nolan, G. P., & Plevritis, S. K. (2011). Extracting a cellular hierarchy from high-dimensional cytometry data with SPADE. Nature Biotechnology, 29(10), 886–891. https://doi.org/10.1038/nbt.1991
- Van der Maaten, L. (2014). Accelerating t–SNE using Tree–Based Algorithms. Journal of Machine Learning Research.
- Van Gassen, S., Callebaut, B., Van Helden, M. J., Lambrecht, B. N., Demeester, P., Dhaene, T., & Saeys, Y. (2015). FlowSOM: Using self–organizing maps for visualization and interpretation of cytometry data: FlowSOM. Cytometry Part A, 87(7), 636–645. https://doi.org/10.1002/cyto.a.22625
* For research use only. Not for use in diagnostic procedures.


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Cardiovascular Risk Factors in Marietta, GA
According to the American Heart Association, half of all people who are diagnosed with congestive heart failure never live more than five years past their diagnosis. This makes heart disease a very scary and serious condition. More troubling, however, is that heart disease, on the whole, is largely preventable.
Early detection and lifestyle modification are key.
Congestive Heart Failure and Heart Disease Risk Factors
These are the leading contributors to developing some form of heart disease whether it be congestive heart failure, coronary artery disease or even a heart attack.
- Hypertension
- Smoking
- Diabetes
- Lack of exercise
- Stress
- Fatigue
- Poor diet, including high sodium
- High Cholesterol
- Obesity
- Age: with age comes physiological changes in myocardial tissue increasing the risk of heart disease
- Men have a greater risk of developing some form of heart disease, however once women hit menopause the odds even out and both genders are at risk.
- A family history of heart disease is a major contributing factor to developing cardiovascular issues.
Other Signs and Symptoms of Heart Disease
While the well-known signs of chest pain, arm numbness, and difficulty breathing are hallmarks of severe cardiovascular issues-and warrant immediate medical attention-here are some overlooked signs that may help you detect heart failure early:
- Clubbing: This is not hitting the town, partying at the latest after-hours hotspot. Clubbing refers to a gradual malformation of the nailbeds on fingers and toes, typically the thumb and forefinger first. Look at where your nails meet your finger near the cuticle. From a side profile, the angle where the nail meets the finger should be about 180-degrees, or straight across. However, if the angle is greater than 180-degrees, this can be a sign of impending heart failure (or pulmonary issues as well). From the side profile again, if-between your knuckle and tip if your finger-the area where your nail connects is elevated, forming a rounded mound of sorts, this is clubbing.
- Hypertrophic osteoarthropathy: If you feel pain and burning in your extremities (lower legs, fingers, toes, hands, and arms) which is typically worse at night, hypertrophic osteoarthropathy may signal the onset of heart disease and eventual heart attack.
- Peripheral edema: This is the swelling of feet and lower legs/ankles. Fluid retention could be a result of poor circulation, caused from a decrease in efficiency of heart function.
- Baldness: Male pattern baldness (baldness that originates at the crown) has been connected to heart failure.
- Xanthomas: If you have yellowish spots under your skin, including your elbows, legs and arms, these fat deposits known as xanthomas could be a result of heart issues and cardiovascular disease.
- Gum disease: While the mouth is a long way from the heart, gum disease, and gum bleeding are often signs of cardiovascular risk and diminishing heart health.
- Tissue edema with weight gain: Weight gain as a result of fluid retention and swelling can be a major red flag, signaling cardiovascular disease.
- Cataracts: As a result of atrial inefficiency to the eye, cataracts may form due to an underlying heart disease like congestive heart failure.
- A night cough: If you have a cough that worsens at night, especially when lying down flat, this could be a sign of heart failure as well.
- Frequent urination: Decreased blood flow, especially the kidneys will reduce their operating efficiency, resulting in your need to urinate more frequently.
Official Tests to Determine Heart Failure
While the signs and symptoms listed above can help with early detection, official medical tests will still be required to confirm a diagnosis of heart issues and determine their severity. An EKG, blood test , stress test, echo, chest X-ray, and an angiogram all have a use in diagnosing cardiovascular system issues.
However, before you reach the diagnosing stage-as the adage goes-an ounce of prevention is worth a pound of cure. It's never too late to improve your heart health, even if you've been diagnosed with a condition like congestive heart failure. It's best to check with your doctor first about the right treatment methods for you, but lifestyle changes, diet improvement, and exercise can make a significant impact on your cardiovascular health.
Regardless of the treatment option, you decide to choose, if you've been diagnosed with a heart condition or simply want to prevent a future one, doing something today is the first step.
Request more information about cardiovascular risk factors today. Call (770) 674-6311 or contact Natural Medical Solutions Wellness Center online.
Natural Medical Solutions Wellness Center
1130 Upper Hembree Rd.Roswell, GA 30076
(770) 674-6311
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The SHOT Team is pleased to announce our second Webinar of 2021 covering the Haemolytic Transfusion Reactions (HTR) chapter of the Annual SHOT Report 2019, with a look at hyperhaemolysis cases and patients with haemoglobinopathies.
This 1-hour webinar is free to access and will include:
- A 40 minute presentation by the SHOT Team and the SHOT HTR Working Experts Dr Anicee Danaee, Dr Tracey Tomlinson and Dr Joseph Sharif covering the key highlights from the HTR chapter of the 2019 Annual SHOT Report
- A 20 minute interactive Q&A session
- An excellent opportunity for CPD through reflective practice
- This webinar will be relevant to all health care professionals involved in transfusion Laboratory Staff, Clinicians, Transfusion Practitioners and Haemovigilance Specialists, as well as Quality Assurance Professionals
Register for the event here: https://nhsbt.zoom.us/webinar/regist...RYa_p2f6s1tm_Q
The Zoom link to the live event is contained in the registration email.
You can also contact us directly to register by e-mailing firstname.lastname@example.com
* *The event will start promptly, please join at least 5 minutes prior to the start**


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"Don't Tell The Boss You Saw Me Here"
Last weekend I stopped at a seasonal garden center in the parking lot of a strip mall in Berwyn, IL. The little garden center is located in a corner of the Cermak Plaza that has the strangest collection of public art I've ever seen and most of it has been there since I can remember. The exception being the sculpture that was made out of trash that was torn down a couple of years ago.
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At a press conference on Friday, the Boulder Police Chief and the district attorney for Boulder County laid out the next steps in the laborious process that lies ahead in bringing justice to the victims of Monday's mass shooting at a local grocery store.
Among that work includes determining a motive, something Police Chief Maris Herold and District Attorney Michael Dougherty said they were still working through.
"The family of victims are desperate to know the motive (of the shooting)," Dougherty said Tuesday. "Whether or not we'll be able to determine that remains to be seen."
When asked directly if international terrorism could have led to the shooting, Herold noted that the FBI and state agencies were doing a "deep dive" on the suspect's background but had no specific updates to share.
She added that questions of the timing and location of the shooting will be "haunting for all of us" until authorities can determine a motive.
Ten people were killed during the shooting at the King Sooper's grocery store, including a Boulder Police Officer. According to authorities, his actions and the actions of other police officers helped save lives that day.
Dougherty added that police responding to the scene faced a "significant amount of gunfire" and put their lives at risk to save others.
Herold confirmed on Friday that the one officer who exchanged gunfire with the suspect, 21-year-old Ahmad Al Aliwi Alissa, had been placed on administrative leave, per department policy. She added that an independent, multi-agency panel will review the facts of the case before determining whether his use of force was justified.
Authorities did not provide much insight into the current state of their investigation, citing the defendant's right to a fair trial. But they noted that much work is still ahead of them.
Herold described the "vastness" of the crime scene and the meticulous work of identifying every shot fired. She noted that it's unlike any other crime scene she's encountered in her career.
The press conference came one day after Alissa, made his first appearance in court.
During that court appearance, the suspect was formally charged with 10 counts of murder and one count of attempted murder. Appearing in a wheelchair due to a bullet wound suffered during the incident, the suspect did not speak outside of confirming to the judge that he understood his rights.
At that hearing, attorneys for the suspect requested her client's mental health be evaluated.
"We cannot begin to assess the nature and depths of Mr. Alissa's mental illness until we have the discovery from the government," Colorado Public Defender Kathryn Herold said Thursday.
District Attorney Michael Dougherty did not object to that motion on Thursday. During Friday's press conference, he noted that he plans to file additional charges of attempted murder against the suspect in the weeks ahead.
The judge then said the status conference would be set in the next 60 to 90 days.
Several vigils honoring the victims of Monday's shootings have been held in Boulder throughout the week.
Read more about the victims of Monday's shootings here.


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Our downloadable cut files are optimised for use with Cricut, Silhouette and Brother Scan n Cut machines, but will work with all electronic cutting machines.
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Celtic Christmas
Classic (and new) seasonal Celtic tunes
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Orchestral and vocal solo holiday music
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A Smooth Jazz celebration of Jewish holidays
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An all-instrumental mix of Smooth Jazz holiday favorites
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A celebration of Hugh Martin and Ralph Blane's 1944 Christmas classic
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A 50/50 mix of Country hits and holiday carols from Country stars like Reba McIntire, Darius Ruck...
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Classical favorites mixed with beautifully performed holiday pieces
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Half Smooth Jazz, half smooth holiday tracks, all good!
One Song Radio: "Blue Christmas"
Featuring dozens of different versions of Elvis Presley's beloved 1957 hit
One Song Radio: "White Christmas"
The Irving Berlin classic in multiple versions -- like snowflakes, each one different!
Curator's Choice: Gary's Christmas Card
Gary puts together the Magic Sunny Lite Hits channels. Here's what he's enjoying at the holidays.
Curator's Choice: Jon's Family Holiday
You can enjoy the same holiday music that Jon, our Classical music programmer, will enjoy with hi...
Curator's Choice: What the Holidays Mean to Jack & Yvonne
Jack is our man behind the Christian channels. He's sharing his and his wife Yvonne's playlist of...
Curator's Choice: Barb's Holiday Greetings
R&B curator, Barbara, brings her love of soul music to this collection with her favorite Christma...
Fifty/Fifty Celtic + Celtic Christmas Blend
The second-best Irish "half & half," a 50/50 concoction of Celtic and Holiday cheer
Songs of Santa
Our listeners' favorite songs about Saint Nicholas from artists like Eartha Kitt, Harry Connick J...
Definitive Versions Christmas
The critics' choice for the single best recording of 100 holiday classics.


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Mining Cable Type G & GG-C. 200' 2 AWG Aluminum XLP USE-2 RHH RHW-2 Direct Burial Cable Black 600V. 2 AWG Aluminum XLP USE-2 cables are primarily used for power distribution, in raceways in accordance with National Electrical Code, except that the cable is not rated for use in cable tray. USE-2 is approved for direct burial and for service entrance below ground. USE-2 cables may be used in wet or dry locations, with conductor temperatures not exceeding 90°C.
Compact Soft Drawn Bare 8000 Series Aluminum, with Class B stranding per ASTM. Cross-Linked Polyethylene insulation XLPE - Abrasion, heat and moisture resistant. NEMA WC-70 - ASTM B800 - ASTM B801 - RUS Accepted - Sunlight resistant. Picture May Not Represent Actual Cable and Is Only Used As A Representation Please Refer to The Specifications For Product Accuracy. Signature required for all deliveries. Feedback will be left as soon as feedback is left for us. This item is in the category "Business & Industrial\Electrical Equipment & Supplies\Wire, Cable & Conduit\Wire & Cable\Multiple Conductor Cable". The seller is "bobdraper85" and is located in this country: US. This item can be shipped worldwide.


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Apology Letter for our Customers
Dear customers,
Please accept our sincere apologies for the inconvenience you may have recently experienced in respect to the technical hitch and SPAM attack both on our e-mail and web-site. We regret that many letters were lost, many questions of yours weren't answered, many orders you placed weren't received by our sales manager.
At Gymnastics-Fantastic, we take pride in ensuring our customer's satisfaction. Unfortunately, we did not meet your expectations. Upon thorough review of the situation, we've made arrangements for our web-store system.
As a testament to our strive for perfection, we have taken steps to ensure that this will never happen again by improving also our e-mail and web-site safety. I do assure you that now we work with maximum performance.
We highly appreciate your relationship with Gymnastics-Fantastic and are committed to providing you with the highest level of service simply because our customers deserve the very best. If you have any further questions or comments regarding this matter, please feel free to discuss it with us at email@example.com or firstname.lastname@example.com (this is my e-mail and it's an ironclad guarantee that your letter will be resent to the sales manager)
If we don't answer your question or order during 24 hours, please, write us one more time. We're doing our best to satisfy any request from our customer and provide every gymnast in the world with a beautiful competition leotard!
We hope for your understanding and further cooperation. Thank you for choosing our company!
Yours in service,
Chief Web-Store Manager
- Don't forget to get some girls gymnastics shoes in reserve.
04 Sep 2015 at 07:00
18 Oct 2012 at 07:00
20 Apr 2017 at 20:13
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OuterVision® Power Supply Calculator
Real Power Consumption
OuterVision Power Supply Calculator is the most accurate PC power consumption calculator available and is trusted by computer enthusiasts, PC hardware and power supply manufacturers across the Globe. Are you building a modern gaming PC, low power HTPC media server, or maybe you need to figure out power requirements for a rack in a data center? We've got you covered - OuterVision PSU Calculator will help you to select a suitable power supply unit and even Uninterruptible Power Supply (UPS) for your system. Building cryptocurrency mining rig? Check our Mining Rig Builder tool.
Video Card Overclock
Basic version of the OuterVision Power Supply Calculator allows users to quickly estimate power consumption with minimal selection of PC parts. On the other hand, our Expert, more advanced version of the PSU Calculator greatly extends the ability to select various PC parts and components, adds CPU and Graphics card overclocking, and allows consumers to calculate PC energy consumption, compare PSU efficiencies, and ultimately project energy cost.
Recommended Power Supply
What power supply do I need? To answer this question, OuterVision PSU Calculator analyzes dozens of power supplies per each certification category, compares their efficiency, unit price, computer power consumption, overall PC energy cost, and payback period. Based on these analysis, we provide recommendation to our users with the best bang for the buck power supply per each certification category. The user has an option to compare the recommended power supplies, adjust electricity cost to see PC's overall energy cost per year, and select the right power supply that fits their budget.


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- Size: 91″ x 91″ x 42″ | 231 x 231 x 96 cm
- 7 seats
- 65 jets
- Capacity: 505 gal | 1,911 L
- Dry Weight: 1050 lbs | 476 kg
- Pumps: (5) 3.0 BHP 1-Spd
Pumps are rated at Brake Horse Power (BHP). BHP is a maximum value measured by the motor manufacturer with no load applied. Dry weight is shipping weight, including package and wooden runners. Dimensions, capacities and weights are approximate. Specifications are subject to change without notice. Spa images have been digitally rendered and may or may not show optional features.
The innovative patented DIRECTFLOW Personal Control®Switchless Motor and Direct Plumbing System combined with the VARIABLE FLOW CONTROL (VFC) provides each user the ultimate individual control and hydrotherapy experience.
- AIR CONTROL – Air pressure adjustment
- PUMP SWITCH – Turn individual seat pump on/off
- Variable Flow Control – High or low jet pressure
• Warranty:
– Lifetime Structure
– 7-Year Surface
– 5-Year Parts, Labor, Plumbing and Electrical
– 2-Year on Monarch Cabinets
• Satin SS Helix and Hydro Extreme Jet Design
• Satin Trimmed Accent Package:
Passivated Satin Stainless Steel Jets, Weir Gate and LED Lighting
Anodized Control Bezels, Waterfall Covers and Cabinet Trim
• Morbern 5 in. Maximum Tapered ASTM Cover
• Full Foam with PermaBase ABS Pan
• WhisperPure™ Circulation System with Microfilter
• Monarch Cabinet
• DynaBrite LED:
(1) 5 in. LED Light, (7) 2 in. LED Lights, (4) Cabinet Corner Lights (N/A with Rock Cabinetry), Cup Holders,
Pillows and Waterfall (Pelican Bay, Piper Glen, Dove Canyon)
(1) 5 in. LED Light, (6) 2 in. LED Lights, (4) Cabinet Corner Lights (N/A with Rock Cabinetry), Cup Holders,
Pillows and Waterfall (Quail Ridge)
• DirectFlow® Personal Control
• Advanced Digital Touch Control System:
Color LCD Panel, WiFi Ready (Module Required), Multi-language Interface
• Titanium Heater: 5.5 kW North America (60 Hz), 3.0 kW Export (50 Hz)
• Crystal AOP™ Brilliance: (FROG®@ease™ with SmartChlor™* & Crystal AOP™)
• 9 in. Waterfall: (2) Pelican Bay, (1) Piper Glen, Dove Canyon, Quail Ridge
• Stereo System:
1) Premium Infinity® Sound System by Harman:
Bluetooth Receiver, AM/FM, USB, Aux-in
(2) Pop-up Speakers, (2) 6.5 in. Exterior Speakers, Power Subwoofer
2) bba2™ Audio
Fully Integrated Bluetooth Audio Streaming
(2) Pop-up Speakers, Subwoofer
• Diamond AOP™
• Nature2® Cartridge
• Pump/Heater Valves
• Worldwide WiFi App Module
Download app for smart device to control hot tub wirelessly
• ProGuard 6 in. Maximum Tapered ASTM Cover
• Rock Cabinetry
* Where allowed by law
Individual DirectFlow™ Control Manages Hydrotherapy at Each Seat
• One Pump Per Seat | Increases efficiency by supplying optimum power and maximum reliability.
• One Control Per Seat | Each spa seat occupant has their own individual control.
• Direct Water Flow | Water flows directly from the pumps to the jets with no diverters, valves or elbows resulting in maximum output with minimum resistance.
Variable Flow Control – Per Seat
• Rotate to High or Low | Rotate to "HI" for a full force massage or to "LO" for a more relaxed massage.
• Full Range | Turn the VFC to any massage power, air pressure and water flow desired, in-between low and high.
• Energy Efficient | When the VFC is turned to low the pumps are using 40% less power consumption than a traditional spa with diverters.
More Power to Control Energy Efficiency – Individual Seat Pumps
The power of massage can be individually regulated by the sophisticated Platinum Elite design and engineering of individual seat pumps, direct plumbing and separate control pads – easily and economically.
No other spa can give you the DIRECTFLOW™ personal control!


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Get Involved
Get Involved!
When the Founding Fathers enacted the Electoral College in 1789, they created a system that made a great deal of sense at the time. Communications technology, access to transportation and voter education were all very poor, limiting the ability of the public to make well informed judgments about candidates.
Now 216 years later, candidates have both the technology and the transportation ability to campaign in all areas of the country, making the voting population well informed and capable of choosing its own president. Although times have changed, the Electoral College has remained the same, becoming a source of disenfranchisement over the years, rather than a source of stability.
This archaic 18th century election system will not merely disappear. We need you to create public awareness and pressure your legislators – local, state and national – to support direct election for the president. Find out all of the ways you can help!
Join FairVote Action's community of democracy activists and take action now to transform democracy.
Support FairVote!
Make a contribution to FairVote through our donation page.
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Lease Revenue Bonds
Featured Links
Proposition 1B
Bond Accountability
Follow-Up Accountability (Audit)
Proposition 1B includes 16 distinct programs or accounts, each having a unique intent and involves a specified set of expectations or desired outcomes. Some are new programs, but many provide additional funding to augment or supplement existing programs with already established guidelines and criteria. Some programs involve a project selection and approval process, while others refer to apportionments as grants or disbursements. Responsibilities for program implementation and delivery also vary with each program. The California Transportation Commission (CTC) and the California Department of Transportation (Caltrans) are identified as the administrative agencies for most of the programs, while the remaining programs fall under the responsibility of the California Air Resources Board (ARB), the California Emergency Management Agency (Cal EMA), or the Department of Finance (DOF). Recipient agencies vary greatly from one program to another, and include State agencies, regional transportation planning agencies, local entities such as counties and cities, transit districts, rail operators, ports, school districts, or others. An accountability plan specific to each of the programs, conforming to the three part accountability structure identified in Executive Order S-02-07, is developed by the administrative agency responsible for the program's implementation or delivery.
To view program specific Follow-Up Accountability Plans, select one of the links below or return to the Proposition 1B overview page.


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TRW LMR (Low Maintenance Rifle) in its final shape, right side, with attached M6 bayonet
TRW LMR (Low Maintenance Rifle) in its final shape, left side, with 30-round magazine attached
Caliber: 5.56x45mm M193
Action: Gas operated, locked by rollers
Overall length: 870 mm
Barrel length: 493 mm
Weight: 3.63 kg loaded
Rate of fire: 450 rounds per minute
Magazine capacity: 20 or 30 rounds
The Low Maintenance Rifle project was initiated by US Army as a result of field experiences, gained through Vietnam war. The standard issue M16 rifle was found to be very sensitive for improper maintenance, and it was decided to look for an infantry rifle, which could work with much less attention paid to routine maintenance, cleaning and lubrication. The one possible niche for such weapons was the "insurgency weapon" for various pro-US (or anti-communist)movements in the third world countries. Development of the LMR project was handled to the US company Thompson-Ramo-Wooldridge (TRW) Inc, and work commence din 1971. Several prototype rifles were built between 1971 and 1973, when US Army decided to stop funding for LMR, as the Vietnam war was over, and M16A1 rifle was considered to be mature enough.
It must be noted that at least one LMR prototype was built for 5.6mm XM216 ammunition that fired saboted sub-caliber flechette projectile. However, most of LMR rifles were built for then-standard 5.56×45 M193 ammunition.
Required "low maintenance" was achieved by using new and improved corrosion-resistant coatings for all metallic parts, as well as thin layers of dry lubricants applied to all moving parts. The number of parts was also decreased, compared to the M16 rifle. TRW LMR was simple in use and could be disassembled and re-assembled without any tools.
TRW Low Maintenance Rifle is gas operated weapon that uses long-stroke gas piston, located at the right side of the barrel. Gas port is located near the muzzle, and gas block serves as a base for front sight. The barrel is enclosed in tubular metallic handguard with oval cooling slots. TRW fired only in full automatic mode, andfrom open bolt. Locking is achieved by a pair of rollers, that engage respective cuts in the barrel extension when in battery. Unlike most other roller-locked designs, LMR bolt had rollers at the top and the bottom, not at left and right.Charging handle was attached to the gas piston rod, and when turned down, locked the bolt in forward position for added safety. Trigger unit and pistol grip were borrowed from US M60 machine gun; manual safety was located at the left side of pistol grip. Magazine (of standard M16 pattern) was inserted horizontally from the left; empty cases were ejected to the right through the ejection port, which has dust cover that opens and closes automatically as gun is fired. Since the LMR was built into so called 'straight' or 'in-line' layout, open sight were put onto high bases. Front sight was of post type, rear sight was a flip-up diopter with two apertures. LMR comba tcapabilities could be further enhanced with addition of light, removable bipod and M6 bayonet, which was attached at the top of gas cylinder, to the right of the barrel axis.


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Key changes from May's report:
- Upgrade Value from Neutral to Positive, Downgrade Growth to Negative from Neutral.
- Upgrade Industrials from Neutral to Positive, Downgrade Technology to Neutral from Positive
- § Upgrade Developed International from Negative to Neutral, Downgrade Emerging Markets to Neutral from Positive.
We expect stocks to continue to grow into their valuations on the back of tremendous earnings momentum as the economy fully reopens, vaccine distribution continues, COVID-19 cases plummet in the United States, and stimulus spending continues to work its way through the economy. As economic growth accelerates, we believe conditions will remain favorable for cyclical value stocks to continue their recent run of outperformance.
Our S&P 500 earnings per share forecast for 2021 is $187.50 to $190, and our 2021 year end S&P 500 fair value target range is 4,400—4.450. After a nearly 90% gain since the March 2020 lows, the S&P 500 may be due for a pause, consistent with the second years of post-WWII bull markets that were characterized by increased volatility. Key risks include COVID-19 spread outside the United States, inflation, rising interest rates, higher taxes, and geopolitics.
- Our equities recommendation remains overweight. We continue to favor stocks over bonds based on our expectation for a very strong economic and earnings recovery in 2021, supported by the full reopening of the U.S. economy and massive fiscal and monetary stimulus.
- As the economic recovery progresses in 2021, we would expect cyclical value stocks to lead. Our favored sectors—mostly on the value side of the ledger—include financials, industrials, and materials.
- Our positive view of small caps is supported by the early-stage bull market and economic expansion and strong earnings. Valuations appear reasonable and recent weakness creates a potentially attractive entry point.
- After upgrading our international developed view and downgrading emerging markets (EM), our regional views are aligned at neutral.
- U.S. stocks enjoy a better economic growth outlook than developed international, but better value-style performance may enable international stocks to match U.S. returns.
- Geopolitical and regulatory threats and a shrinking economic growth advantage have left us with less conviction on EM.
- We continue to recommend an underweight allocation to fixed income. Although we've seen a big move higher in rates this year, Fed policy and manageable inflation may limit the risk of further large moves in the near term. Higher rates may still put some pressure on bond returns while economic improvement may help support equities going out a full year.
- We favor a blend of high-quality intermediate bonds that is underweight U.S. Treasuries with an emphasis on short-to-intermediate maturities and sector weightings tilted toward mortgage-backed securities (MBS).
Click here to download a PDF of this report.
This material has been prepared for informational purposes only, and is not intended as specific advice or recommendations for any individual. There is no assurance that the views or strategies discussed are suitable for all investors and they do not take into account the particular needs, investment objectives, tax and financial condition of any specific person. To determine which investment(s) may be appropriate for you, please consult your financial professional prior to investing. Any economic forecasts set forth may not develop as predicted and are subject to change.
Stock investing involves risk including loss of principal. Because of their narrow focus, sector investing will be subject to greater volatility than investing more broadly across many sectors and companies. Value investments can perform differently from the market as a whole and can remain undervalued by the market for long periods of time. The prices of small and mid-cap stocks are generally more volatile than large cap stocks. Bonds are subject to market and interest rate risk if sold prior to maturity.
Bond values will decline as interest rates rise and bonds are subject to availability and change in price. Corporate bonds are considered higher risk than government bonds. Municipal bonds are subject to availability and change in price. Interest income may be subject to the alternative minimum tax. Municipal bonds are federally tax-free but other state and local taxes may apply. If sold prior to maturity, capital gains tax could apply. U.S. Treasuries may be considered "safe haven" investments but do carry some degree of risk including interest rate, credit, and market risk. Bond yields are subject to change. Certain call or special redemption features may exist which could impact yield. Mortgage backed securities are subject to credit, default, prepayment, extension, market and interest rate risk.
Credit Quality is one of the principal criteria for judging the investment quality of a bond or bond mutual fund. Credit ratings are published rankings based on detailed financial analyses by a credit bureau specifically as it relates the bond issue's ability to meet debt obligations. The highest rating is AAA, and the lowest is D. Securities with credit ratings of BBB and above are considered investment grade. Duration is a measure of the sensitivity of the price (the value of principal) of a fixed-income investment to a change in interest rates. It is expressed as a number of years.
Alternative investments may not be suitable for all investors and should be considered as an investment for the risk capital portion of the investor's portfolio. The strategies employed in the management of alternative investments may accelerate the velocity of potential losses.
Commodity-linked investments may be more volatile and less liquid than the underlying instruments or measures, and their value may be affected by the performance of the overall commodities baskets as well as weather, geopolitical events, and regulatory developments. The fast price swings in commodities and currencies will result in significant volatility in an investor's holdings.
Investing in foreign and emerging markets securities involves special additional risks. These risks include, but are not limited to, currency risk, geopolitical risk, and risk associated with varying accounting standards. Investing in emerging markets may accentuate these risks. All information is believed to be from reliable sources; however, LPL Financial makes no representation as to its completeness or accuracy.
Earnings per share (EPS) is the portion of a company's profit allocated to each outstanding share of common stock. EPS serves as an indicator of a company's profitability. Earnings per share is generally considered to be the single most important variable in determining a share's price. It is also a major component used to calculate the price-toearnings valuation ratio.
Gross Domestic Product (GDP) is the monetary value of all the finished goods and services produced within a country's borders in a specific time period, though GDP is usually calculated on an annual basis. It includes all of private and public consumption, government outlays, investments and exports less imports that occur within a defined territory.
All index data from FactSet.
For a list of descriptions of the indexes referenced in this publication, please visit our website at lplresearch.com/definitions.
Securities and advisory services offered through LPL Financial (LPL), a registered investment advisor and broker/dealer (member FINRA/SIPC). Insurance products are offered through LPL or its licensed affiliates. To the extent you are receiving investment advice from a separately registered independent investment advisor that is not an LPL Financial affiliate, please note LPL Financial makes no representation with respect to such entity.
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