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BURLINGAME, Kan. � The grave-site of three Columbia residents who were slain by their husband and father stands just a few hundred yards from the site where their lives were taken. The shared headstone lists their separate names and birthdates, but large, bold letters across the bottom spell out a date they share for eternity: �November 28, 2009.�
An Osage County jury yesterday convicted former Columbia Water and Light Director Kraig Kahler, 48, of capital murder for shooting his wife, Karen Kahler, 44; their daughters, Emily, 18, and Lauren, 16; and Dorothy Wight, 89, Karen Kahler�s grandmother, on that date at Wight�s home in Burlingame.
Few who followed the trial seem to have bought the now-convicted capital murderer�s insanity defense, but they also said no sane man could kill his children the way Kahler did.
�He�s possibly a little insane,� Burlingame resident Troy Prescott said. �I don�t see a reason to ever do what he did.�
Prescott, like many residents of the small rural community of Burlingame, has been keeping up with the capital murder trial through media reports. The quadruple slaying shocked local residents and, to some, altered their small-town sense of security.
Although many residents are open to speaking about the slayings, others who knew Wight, a longtime resident of Burlingame, are angry.
�I knew the family a little bit, and I prefer not to talk about it,� said David Prescott, Troy Prescott�s father. �That was straight-out murder. � He deserves the death penalty.�
Kahler�s sentencing phase begins Monday. The same jurors who returned a guilty verdict on all counts will decide his fate � the death penalty or life in prison without parole.
Death by lethal injection might be too easy a penalty given the crimes, Burlingame resident Arin Zirkle said.
�It�s too easy to just say you�re insane,� she said. �If he was angry enough, he could have done it. � I don�t feel it�s fair to sentence� him �to death. I don�t feel like it�s enough punishment.
�If he is insane, he�s insane now because he has to deal with the fact that he killed them.�
As in Burlingame, some friends of the victims in Columbia yesterday were glad to hear news of the verdict.
�I�ve been checking the newspaper� website �all day,� said Shay Gann, a former member of Karen Kahler�s boot-camp classes she led at Columbia�s Activity & Recreation Center. �I yelled out, �He has been found guilty,� and started texting members of our boot camp.�
She said she didn�t buy the insanity defense, either, but she also questioned how any sane person could murder his children.
Although reading the things the defense alleged against her friend was difficult, Gann said, now justice can be served.
�If given the death sentence, and it is actually carried out, he will still die in a much more humane way than his family did,� she said.


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As plastic surgery has evolved over the years, Phalloplasty procedures has come a long way to where it is today. Complicated ridden multistage tube pedicles which were popular earlier are now replaced by forearm flaps with the advent of microsurgery. It is now also quite possible to reconstruct aesthetically pleasing glans as well which meets both aesthetic & functional desires of patients. However, there still are complications encountered in the reconstructive effort which includes urethral stricture, urethral fistula, flap failure & stiffener related problems.
It is one of the most enduring dreams in medicine so as to possibly replicate or replace human organs which have been lost to disease or trauma. Phallus, which is symbolic of manhood, has however received far less attention than the breast, heart, liver or kidney, possibly just because one can survive without a penis. Another reason which is possible is that function of this organ almost appears to be magical by the way it is able to respond to changes in emotion & environment. Moreover, unlike many other organs, cadaveric transplantation seems to be desecrating. With plastic surgery trying to replace like with like, real phalloplasty would only appear to be a dream. Nevertheless, this, in any case, does not undermine the importance of efforts to reconstruct as perfect a phallus as is possible with materials that are available; perfection being not just in form but in function as well.
Though most phalloplasty cases predominantly consist of gender reassignment cases, indications for this plastic procedure are not limited to this group of patients anyway. Some main indications to opt for phalloplasty procedures are listed below.
Patient's perception of normal form & function is extremely relevant. Patients are generally looking for a scrotum, glans, rigidity & an aesthetically appealing appearance while being nude or wearing a tight swim suit.
Many different techniques have been utilized over the years so as to reconstruct the penis in order to achieve the goals mentioned above. With advancements in plastic surgery, especially with the advent of microvascular surgery, the bar for expected goals has constantly been rising. Various methods utilized for phalloplasty surgery are mentioned below.
Prior attempts at phalloplasty were only directed towards the reconstruction of the form of male genital organ & urethral reconstruction was subsequently added as a secondary goal. Since the construction of neourethra has frequently led to strictures & fistulas, some surgeons have even abandoned progressive attempts. Understanding of patient's requirements however improved & urethroplasty was finally included as a primary goal of phalloplasty. Different techniques have subsequently been described for reconstruction of the urethra. Prefabrication, prelamination, tube-in-tube or even separate flaps have been successfully used for reconstruction of the pendulous urethra. Local vaginal or labial flaps, separate flaps, free skin or mucosal grafts & extra long urethral component of phalloplasty flap have been described for reconstruction of fixed urethra. However, improved outcome has been achieved by use of prelamination techniques, but they are found to increase the number of stages required to complete the phalloplasty procedures.
Reconstruction of glans penis has now become an important portion of phalloplasty patient's wishes for a normal & aesthetic looking penis. This is a procedure which involves raising a circumferential de epithelializing skin flap at level with the proposed coronal ridge which is subsequently rolled up while the free edge of the flap is sutured to its own base in order to form a ridge. Following this, the raw area is covered with graft. Split-thickness skin graft is able to produce a normal-looking coronal sulcus than what a full-thickness skin graft could. While providing the best results, this technique has been regarded as an ideal method for coronal ridge & sulcus construction.
Regarded as one of the most complex reconstruction procedures which plastic surgeons are called upon to perform, total phalloplasty involves replicating form & function of a phallus which is truly unique. Along with this fact, sub-optimal material availability & emotional overlay which is associated with phallus reconstruction, this task assumes a Herculean responsibility for plastic surgeons. Therefore, it is no wonder that phallus reconstruction is fraught with a plethora of complications. Although survival of the flap & appearance is primarily the first requisite to successful phallus reconstruction, it is also the function which is both urinary & sexual, which will eventually determine success or failure.
However, phalloplasty procedures has no clear cut indications & a homogenous population of patients. Even decision for using a particular flap cannot be based upon rates of flap survival & functional outcomes alone. Indication for gender reassignment or for loss can also influence flap selection as visible forearm donor site can stigmatize individuals who are transgender. Plastic surgeons starting out with such challenging reconstructions are advised to train with experts before picking up a particular technique. Radial forearm which portrays a reliable anatomy, long pedicle with good caliber of vessels & pliable sensate skin should primarily be considered as the first choice of treatment for most indications. This has been the gold standard for phalloplasty procedures & various other modifications along with this can prove very useful in particular situations. Flap survival rates are also very high with this technique except for the hair-bearing nature fistulae & strictures which are quite common. Moreover, visibility of donor site is also a serious drawback. Other options which find considerable usage is pedicled ALTF which provides a large amount of skin. Moreover, donor site is also well hidden & total flap loss is unlikely since it is a pedicled flap. This is usually thick but can also be successfully thinned by application of liposuction. Sensory recovery in this phalloplasty technique is, however, poorer when compared to radial forearm flap.
Development of techniques for phalloplasty has effectively paralleled the evolution of plastic surgery, both in terms of popularity & practicability. Tubed pedicled flaps were sensibly followed by RFFF as the standard technique for phalloplasty. Moreover, an osteocutaneous variant of forearm flap & OCFF held several promises including an all-for-one & one-for-all function provided in a single flap. Prefabrications & prelaminations were subsequently introduced in order to reduce rates of urethral complications. Roll of stiffener prostheses was also introduced to assist erection during sexual intercourse. However, the fact is that RFFF method has by far withstood the test of time & is still offering a single-stage technique for total penile reconstruction.
Uncontested space of several conventional free flaps has been effectively challenged in penile implant surgery with the advent of perforator flaps. Similarly, the introduction of ALTF in sensate pedicled format has all of a sudden started replacing RFFF as the best choice of phallus reconstruction. The main advantage of this new-found technique is that the donor site is well hidden within everyday clothing along with no necessity for microvascular expertise required from the surgeon. Earlier, since no sensibility in phallus was expected to be considered, no stiffeners were used in pedicled flaps as these were regarded to be techniques featuring few indications for phalloplasty. However, things have changed & innervated pedicled ALTF has proved to be a workable proposition & has also proved to effectively solve the problem of stiffener retention.
When the cost of healthcare has been rising all around the world, especially in the developed western countries, India has emerged as an ideal alternative providing excellent quality medical procedures at affordable costs for the international patient. Healthcare infrastructure in India is comparable to the best in the world & is composed of globally reputed doctors & surgeons performing all kinds of procedures in internationally accredited hospital facilities. IndianMedTrip is one of the fastest growing medical tourism companies which is based in South Asia & is offering a wide spectrum of low-cost healthcare solutions including plastic surgeries like phalloplasty to people willing to seek medical treatments away from their homeland. Seamless services offered by this company begin with the first telephonic conversation & move on to identifying the best doctor & ideal hospital as per the patient's requirement. Other services provided by IndianMedTrip include assisting in obtaining medical visas for the patient & his/her accomplice, warm reception at the airport on arrival, convenient local transport, comfortable accommodation options including budget, luxury or serviced apartments, scheduled appointments regarding tests & medical procedures with doctors & surgeons, option for availing exotic recuperative vacations at reasonable costs, follow-up evaluation checks & a successful farewell at airport when the patients is heading back home.

subset: commoncrawl


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Jack of All Games Founder David Rosenbaum Joins Solutions 2
Orange County, Calif. - May 4, 2011 -- Solutions 2 GO, LLC ( www.solutions2go.com), a value-added distributor of video games in the United States, announced today that David Rosenbaum has joined the company as Vice President of National Accounts. The company offers full distribution and retail marketing services for video game console and handheld systems, games, accessories, and related products for the Sony, Nintendo, and Microsoft gaming platforms with a key focus on providing retail marketing and promotional merchandising solutions for its vendor and customer partners. Solutions 2 GO, LLC is headquartered in Orange County, California. In his role as Vice President of National Accounts Rosenbaum will manage the company's relationship with several key customer partners.
"David's experience in the industry, from his founding and managing of Jack of All Games to his executive involvement with ZOO Publishing, has given him a well-rounded background and understanding of every part of the video game industry," said Nima Taghavi, Chairman of Solutions 2 GO. "David's extensive knowledge of the industry and his tireless work ethic make him a valuable and great fit for our team. We are fortunate to have him," added Solutions 2 GO CEO, Michael Maas.
Rosenbaum has more than 30 years of experience in the Toy and Video Game Industries. Most recently Rosenbaum served as the President of Zoo Publishing, Inc. from April 2009 through March 2011 and prior to that served in various capacities at Zoo Publishing, including Senior VP of Sales, dating back to July 2006.
In 1989, Rosenbaum founded Jack of All Games, which he sold to Take Two Interactive in 1998, but remained on as President through March 2006.
Rosenbaum received a B.A. from the University of Cincinnati.
"Solutions 2 GO is a perfect fit for my experience and knowledge of the video game industry," said Rosenbaum. "I have admired Michael and Nima and their success for over 15 years and I am proud and excited to be part of the Solutions 2 GO Team."
Solutions 2 GO, LLC specializes in direct-to-store shipments of retail-ready products and displays, merchandise assortment planning, unique retail promotions and product bundling, exclusive products, in-store merchandising and display support, retail channel marketing, vendor managed inventory programs with life-cycle product management, and consumer direct fulfillment services. The company has a fully operational warehouse with pick, pack, and ship capabilities, as well as an array of additional value added services.
About Solutions 2 GO, LLC. (USA)
Based in Orange County, California, Solutions 2 GO, LLC (USA) is a value-added sales, marketing, and logistics company of video game entertainment products for the Sony PSP® system, Sony PlayStation®2 computer entertainment system, Sony PlayStation®3 computer entertainment system, Nintendo WiiT, Nintendo DST, and the Microsoft Xbox 360T video game and entertainment system.
Mika Kelly
Clever communications
|[View All]


text:
I took the Geek, Nerd, or Dork test and I'm not surprised at the finding.
Pure Geek
43 % Nerd, 78% Geek, 39% Dork
For The Record:
A Nerd is someone who is passionate about learning/being smart/academia.
A Geek is someone who is passionate about some particular area or subject, often an obscure or difficult one.
A Dork is someone who has difficulty with common social expectations/interactions.
You scored better than half in Geek, earning you the title of: Pure Geek.
It's not that you're a school junkie, like the nerd, and you don't really stand out in a crowd, like the dork, you just have some interests that aren't quite mainstream. Perhaps it's anime, perhaps it's computers, perhaps it's bottlecaps, perhaps it's all of those and more. Your interests take you to events and gatherings that are filled with people you find unusual and beyond-the-pale, but you don't quite consider yourself "of that crowd."
.: 155 words at 12:38 AM in Quizzy Stuff :: Link :: Pings
:: All the Voices Say... (3)
Show comments here »
Show comments here »
Well your one sexy geek! :)
Posted by Terry at September 6, 2005 04:52 PM
Pure Nerd
91 % Nerd, 43% Geek, 30% Dork
And I color code my spice drawer!
Posted by Pam Heath at September 7, 2005 03:48 PM
Joe Normal
34 % Nerd, 43% Geek, 34% Dork
I guess that's good?
Posted by girl at September 17, 2005 06:27 AM
« Hide comments


text:
What Books From The New Thought Greatest Personality Read?
Mother Teresa– St.Augustine's Memories "For what is faith unless it is to believe what you do not see"
Rhonda Byrne – The Science Of Getting Rich by Wallace Wattles this is the legendary book from which the inspiration for The Secret came!'
Bill Gates -The Master Key System by Charles Haanel- These are the techniques which made millionaires over and over again, before the book basically disappeared about 1933. This is the very book that later re-emerged in Silicon Valley decades later as a 'millionaire-maker' all over again. The legend goes that a certain Bill Gates stumbled upon this book when just a Harvard student and went on to create history.
Henry Ford-Napoleon Hill-Norman Vincent Peale- In Tune With Infinity by Ralph Waldo Trine "Within yourself lies the cause of whatever enters into your
life. To come into the full realization of your own awakened
inner powers is to be able to condition your life in exact
accord with what you would have it.
Gandhi– The New Testament JESUS Teachings about Faith-Love and Peace.
Bob Proctor– Think and Grow Rich by Napoleon Hill and The Science Of Getting Rich by Wallace D. Wattles.
Thomas Alva Edison– The Spiritual; Laws by Ralph Waldo Emerson and Treatise by Spinoza.
Graham Bell- William Blake.
I will continue adding more to this list.


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text:
Donald Trump's US tax cuts will help lift global growth further over the next few years though the UK looks set to trail behind, according to latest forecasts from the International Monetary Fund (IMF).
The IMF's World Economic Outlook pencils in expansion of 3.9% for the world economy for this year and next – an upgrade of 0.2 percentage points in each case.
But it forecasts growth in the UK at 1.5% in each of 2018 and 2019, lagging well behind the average pace of fellow advanced economies – having led the pack as recently as 2016.
For the year 2019, the outlook represents a 0.1 percentage point downgrade for Britain.
IMF chief economist Maurice Obstfeld said the main sources of the upturn in world GDP were in Europe and Asia, with the US and Canada and a return to expansion for large emerging markets Brazil and Russia – which shrank in 2016 – as well as Turkey.
The body also said recently-implemented US tax cuts would stimulate growth, adding 1.2% to the world's biggest economy by 2020 – though due to the temporary nature of the some of the policies it would lower growth from 2022 onwards.
The IMF said the impact on the US and its trading partners would "contribute about half of the cumulative revision to global growth over 2018-19".
Video: A very good year: Trump and the US economy
Mr Obstfeld said: "As the year 2018 begins, the world economy is gathering speed.
"This is good news. But political leaders and policy makers must stay mindful that the present economic momentum reflects a confluence of factors that is unlikely to last for long."
He warned that "the next recession may be closer than we think" and that it may be harder to deal with than a decade ago given the scale of public sector debt.
UK growth has been slowing in the wake of the Brexit vote.
It had been, together with Germany, the fastest-growing of the G7 economies in 2016.
The pace of growth of the advanced nations was 1.7% in that year and is expected to have advanced to more than 2% for last year, this year and next.
But the UK is forecast to go in the opposite direction, with growth pencilled in by the IMF at 1.7% for 2017 before slipping further.
Last year's slowdown has been blamed on the squeeze on consumers caused by higher inflation – largely the result of the pound's plunge after the Brexit vote – as well as uncertainty over Britain's future outside the EU.
However, survey reports have also pointed to a boost to Britain's manufacturing sector as the weakness of the pound, combined with the strength of the global economy, lifts exports.
The IMF's latest report cited possible trade barriers and regulatory shake-ups as risks to growth as the UK seeks a new relationship with Europe after Brexit and the US renegotiates its free trade treaty with neighbours Mexico and Canada.
A Treasury spokesperson responded to the report by saying: "We are building a Britain that is fit for the future by improving skills, backing innovation and investing in infrastructure to deliver a stronger economy and guarantee a better future for the next generation."
Shadow Chancellor John McDonnell said: "The IMF report out today confirms what others have been saying – the UK economy is not growing as fast as many other advanced economies. It further exposes the effect of the last seven years of Tory economic failure.
More from Business
"Philip Hammond should drop his plans to continue the austerity policies that have weakened our economy. We urgently need a change of direction from the Chancellor. There is no excuse for a 'more of the same' approach.
"The next Labour government will provide the serious investment our country needs, underpinned by our Fiscal Credibility Rule, to build a high wage, high skill economy for the many not the few."
Source: Sky


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by Ron Paul | April13, 2008
Texas Straight Talk
There has been a lot of talk in the news recently about the Federal Reserve and the actions it has taken over the past few months. Many media pundits have been bending over backwards to praise the Fed for supposedly restoring stability to the market. This interpretation of the Fed's actions couldn't be further from the truth.


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Can you guess what my inspiration for today's post was?
Usually at this point there would be a list of items in the background and a pose credit, but obviously, there's absolutely no point as this is just my face. I hope you'll continue reading, however, because I have a bunch of information about the Lotus skin & Eyes used in this image shown below!
All of the head things :
Head : CATWA Magy
Skin : LOTUS.10 Rebecca Skin (tone 02) (Catwa) @ Pocket Gacha
Eyes : CATWA Mesh Eyes
Eyes : LOTUS. Rebecca Eyes @ Pocket Gacha
Gag/Harness : [Cubic Cherry] Innocence gag @ RLD
Hair : Tableau vivant \\ Nouvelle hair (4 options, pulled forward, pulled back & L&R strands) @ Kustom9
All of the informational things :
For those of you who aren't familiar with Pocket Gacha, it's basically a hud-based game of chance that works exactly like in-world gacha machines. A bunch of creators are involved, you grab a hud and rather than teleporting to a laggy sim where you can't move and nothing rezzes and OMG I'M BEING EJECTED BECAUSE MY SCRIPTS ARE TOO HIGH, you sit in the comfort of your platform/house/that one dance club you like, and click through until you find something that sparks your interest. You then select the option to either pay the hud in Lindens, or specially purchased credits, and play the gacha you desire until you a. get the item you want, or b. give up and cry 5ever. Like all gachas there are a set RARE percentage, and with some creators there's the option to purchase a full set without even playing on the machine and trying your luck! For more information you should check their website at the link up there ^^, because I'm faffing and probably explaining super poorly! :P
Lotus has released this deliciously lovely skin & eye set called Rebecca, especially for the March round of Pocket Gacha. Here are some pictures!
This is the Rebecca skin, in CalWL on Ultra grapics in Firestorm viewer. I haven't used any shadows, and there is no shine on my catwa head, also, zero editing in photoshop other than the cropping effects when I popped the pictures together.
Shown here from left to right are the Tone 3 versions available, LOTUS.7 Rebecca Skin (tone 03), LOTUS.8 Rebecca Skin (tone 03), LOTUS.9 Rebecca Skin (tone 03).
This is my favourite tone! It's my humble opinion that the Lotus skins have come on leaps and bounds in this edition, the textures are much smoother, and the detailing across the nose and lips are much improved! I'm a real sucker for a cute button nose, and I don't feel at all let down here. I'm also absolutely in love with these cat-liners. They're sharp and realistic down to the matte effect on my eyelid, and I am doing some serious heart hands!
Shown from left to right are the Tone 2 versions, LOTUS.10 Rebecca Skin (tone 02), LOTUS.11 Rebecca Skin (tone 02), LOTUS.12 Rebecca Skin (tone 02)
Here are the rare skin hud tones.
It's worth mentioning that ALL the skins, whether common or rare come with the body applier huds for Maitreya and Slink, so you can guarantee a match for all of your bits at no extra cost. Honestly, each one of these skins are lovely and I'm ecstatic to be able to add them all to my collection!
And now for a musical interlude until next time. I'm sure you'll find this familiar.
Kinda apt this'd be a skin post, really... don't you think?
*Scampers off to put the lotion on her skin before she gets the hose again*


text:
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In my article titled Bikini body for anyone? You bet your ass! I mentioned that I would write about the true intention of the BMI. This is the single most important thing you should know. The calculation for the Body Mass Index (BMI) was NEVER MEANT TO BE APPLIED TO AN INDIVIDUAL. Yes. I have deliberately added a lot of emphasis on these words. I love the explanation provided by Dr. David Lewis and Dr. Margaret Leitch in their book Fat Planet – The Obesity Trap and How We Can Escape It.
"The statistical technique for comparing people's weight was invented by the eighteenth-century Belgian statistician and polymath Adolphe Quetelet. At the request of the Belgian government, who wanted a quick and easy technique for determining obesity levels among the general population, he created a standardized measure based on an individual's weight divided by the square of their height. It was an assessment technique that, he always emphasized, could and indeed should, only be used to assess groups and never individuals. (My emphasis added here).
This calculation was developed 300 years ago for a specific purpose similar in use as that of insurance companies and actuarial tables based on statistics applied to the overall population. For example, men under 25 have more car accidents than the rest of the population or smokers are more likely to get sick and to die earlier than non-smokers. Insurance rates are based on these tables. However, it doesn't mean that every male under 25 will be in a car accident or that every smoker will be sick and die sooner than a non-smoker. Following this same logic, being placed into one of five categories (under, over, obese, morbidly obese, just right) based on a BMI number is ludicrous.
Here's an example. Take a 6'2" tall body builder weighing 260 lb. His BMI would be 33.3, which, according to the tables, is considered obese. You might be interested to know I'm describing Arnold Schwarzenegger when he was Mr. Universe.
I'm sure you have seen picture of body builders.
There is no fat to be found anywhere. They are all muscle. How can you term these people as obese?
BMI applied to an individual as a measure of fitness is not only irrelevant but downright irresponsible. It doesn't take into account bone structure and density or muscle mass.
In fact, it doesn't take into account so many other factors that lead to whether or not a person is healthy. So, a 5'2″, 125 lb, completely sedentary person with small bones, little fat and on a diet of junk food with a BMI of 24 would be considered "just right" whereas a 5'2″, 137 lb, person who has a little extra around the middle but eats healthy food, exercises regularly, is full of energy but has a BMI of 33 would be considered obese.
The BMI scale does not reflect overall health and, in my opinion, should be abandoned. It assigns a number, thereby giving the illusion of legitimacy and scientific validity but is a poor indicator of real health.
It's time for the BMI to be put to its intended use, classifying populations instead of individuals for policy making purposes. The more personal system for determining individual health and wellness needs to be re-applied. That means, doctors taking time to speak to and assess their patients and treat each based on his or her respective needs.


text:
Tomahawk Board Members and Committees
Tomahawk Regional Chamber of Commerce Board of Directors
President, Andrew Hein-Foley's Tree Service
Vice President, Ron Volz-Outdoor Solutions of the Northwoods
Treasurer, Gladi Felser-Nokomis ATV Club
Secretary, Audrey Cordova-Tomahawk Community Bank
Jesse Bolder-WJJQ Radio
Tricia Hoffman-Scentability and Friends
Chief Al Elvins-Tomahawk Police Department
John Krueger-Krueger Family Funeral Home
Mike Meinheit-Twin Bay Resort
Frank Sus-Sus Convenience Stores
Jeffrey Johnson-Tomahawk Extended Area Partners in Education
The Chamber has various committees and opportunities to volunteer. One of our most active committees is our Tomahawk Ambassadors. The Ambassador group welcomes new businesses to town by hosting ribbon cutting meet and greets. If you are new to town and interested please contact the chamber office between 8-4 on weekdays. If you are interested in getting involved in one of our committees or volunteer opportunities send us an email and we will be in touch [email protected].


text:
- Research article
- Open Access
The impact of RNA sequence library construction protocols on transcriptomic profiling of leukemia
© The Author(s). 2017
- Received: 23 March 2017
- Accepted: 8 August 2017
- Published: 17 August 2017
RNA sequencing (RNA-seq) has become an indispensable tool to identify disease associated transcriptional profiles and determine the molecular underpinnings of diseases. However, the broad adaptation of the methodology into the clinic is still hampered by inconsistent results from different RNA-seq protocols and involves further evaluation of its analytical reliability using patient samples. Here, we applied two commonly used RNA-seq library preparation protocols to samples from acute leukemia patients to understand how poly-A-tailed mRNA selection (PA) and ribo-depletion (RD) based RNA-seq library preparation protocols affect gene fusion detection, variant calling, and gene expression profiling.
Overall, the protocols produced similar results with consistent outcomes. Nevertheless, the PA protocol was more efficient in quantifying expression of leukemia marker genes and showed better performance in the expression-based classification of leukemia. Independent qRT-PCR experiments verified that the PA protocol better represented total RNA compared to the RD protocol. In contrast, the RD protocol detected a higher number of non-coding RNA features and had better alignment efficiency. The RD protocol also recovered more known fusion-gene events, although variability was seen in fusion gene predictions.
The overall findings provide a framework for the use of RNA-seq in a precision medicine setting with limited number of samples and suggest that selection of the library preparation protocol should be based on the objectives of the analysis.
- RNA-sequencing
- Hematological malignancies
- Library preparation
- Ribo-depletion
- Poly-A selection
RNA sequencing (RNA-seq) has become an important technology in the comprehensive analysis of disease transcriptomes and holds great promise for clinical applications including disease diagnosis, therapeutic selection, and precision medicine strategies [1–4]. The technique has been particularly insightful in understanding the pathogenesis and classification of leukemia [5, 6]. For example, it has enabled identification of a wide variety of clinically relevant predictive expression biomarkers [4, 7], fusion-genes and recurrent mutations [8, 9], expressed variants [5], and alternative splicing events [10] in different leukemia types. However, as a relatively new technology, sample preparation protocols and data analysis methods are still in their infancy and require further testing before RNA-seq can be translated to standard clinical practice [2].
Generation of a sequencing library for RNA-seq analysis is a complex, multi-step process and a potential source of significant variation [11, 12]. This process is most commonly carried out using poly-A-tailed mRNA selection (PA) or rRNA depletion (RD) to eliminate rRNAs that are naturally abundant in the sample and which would otherwise dominate the sequence data [13, 14]. However, both of these mainstream methods have their own advantages and limitations. For example, recent studies have noted that the RD protocol captures a wide repertoire of transcripts [15, 16] and works efficiently with degraded RNA [12, 15]. The high number of intron mapping reads in RD datasets may also be advantageous in understanding pre-mRNA dynamics and the post-transcriptional impact of microRNAs [17]. In contrast, Li et al. have reported PA libraries to contain less intronic reads than RD libraries [12] thereby offering a more cost-effective solution for gene expression studies [18]. The PA method also appears to outperform the RD protocol in detecting differentially expressed genes [15, 18]. However, the assessments of different RNA-seq library preparation protocols have mostly relied on non-clinical samples [16, 18–20], emphasizing the need for systematic comparison of library preparation protocols using patient samples. To address this need, our comparative analysis provides recommendations for the application of RNA-seq in clinical or pre-clinical settings with a limited number of samples.
In this study, we tested the performance of PA and RD protocols on samples from acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) patients in a personalized medicine setting. For each patient sample we generated two RNA-seq libraries using the PA and/or RD protocols. We then assessed the effects of the two different protocols on i) expression of protein coding and non-coding RNAs, ii) differential gene expression analysis, iii) pathway analysis, iv) fusion gene detection, and v) expressed variant calling. In addition, we measured the variability introduced by library preparation in technical replicates along with biological replicates and developed metrics applicable in routine medical practice and other small-n settings by integrating RNA-seq data with variant, biomarker, and ex vivo drug sensitivity and resistance testing (DSRT) data. Our analyses showed that PA and RD protocols produced consistent results and that patient heterogeneity represented the largest source of variation. However, the RD method captured more transcriptome features whereas PA outperformed the RD protocol in detecting differentially expressed genes and leukemic markers. Importantly, some of the observed discrepancies were clinically relevant and therefore selection of the protocol is a crucial step in clinical decision-making. Our results are directly relevant for researchers and healthcare professionals aiming to apply RNA-seq in a precision medicine setting to examine transcriptomes of hematological diseases for clinical assessment and indicate that the selection of the library preparation protocol should be guided by the study objectives.
Patient material
The Helsinki University Hospital Ethics Committee has approved the study and collection of samples (permit numbers 239/13/03/00/2010, 303/13/03/01/2011). Bone marrow (BM) aspirates from two AML and two ALL patients were collected after signed informed consent and with protocols in accordance with the Declaration of Helsinki. Mononuclear cells (MNCs) were isolated by density gradient separation from the BM of the patients (Ficoll-Paque PREMIUM; GE Healthcare; Little Chalfont, Buckinghamshire, UK).
mRNA purification and library construction
Total RNA was extracted from MNCs using the Qiagen miRNeasy kit (Qiagen, Hilden, Germany). The kit is capable of isolating all types of RNA from a minimal amount of starting material. Short RNAs (< 200 nt) present in the total RNA were removed prior to preparation of the RNA-seq libraries. Next, RNA was quantified using the Qubit fluorometer (Thermo Fisher, Carlsbad, CA, USA), while the quality of the RNA samples was measured using a Bioanalyzer instrument and RNA nano chips (Agilent, Santa Clara, CA, USA). For the PA protocol, 2.5-5 microgram of total RNA from the ALL 542 and AML 800 samples was then subjected to oligo(dT) selection using the Dynabeads® mRNA Purification Kit (Thermo Fisher) as per the manufacturer's instructions. The RD protocol was carried out from 2.5-5 microgram of total RNA from ALL 542, ALL 668, AML 800 and AML 1867 samples using the Ribo-Zero™ rRNA Removal Kit (Epicentre, Madison, WI, USA) as per the manufacturer's instructions. After PA or RD selection, the samples were purified using Agencourt AMPure XP SPRI beads (Beckman Coulter, Brea, CA, USA) to remove chemical contaminants and short RNAs less than 200 nt in length.
PA and RD samples were further reverse transcribed to double stranded cDNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Thermo Fisher). Random hexamers (New England BioLabs, Ipschwich, MA, USA) were used for priming the first strand synthesis reaction. Samples were prepared for RNA-seq using Illumina compatible Epicentre Nextera™ technology. After limited cycle PCR the RNA-seq libraries were size selected (350–700 bp fragments) in 2% agarose gel followed by purification with the QIAquick gel extraction kit (Qiagen).
RNA sequencing
Each transcriptome was loaded to occupy one third of the lane capacity in the flow cell. The cBot-2 system and TruSeq PE Cluster Kit v3 (Illumina, San Diego, CA, USA) were used for cluster generation, and TruSeq SBS Kit v3-HS reagent kit and HiSeq2000 instrument (Illumina, San Diego, CA, USA) was used to generate paired 100-bp reads according to the manufacturer's instructions. Nextera Read Primers 1 and 2 as well as Nextera Index Read Primer (Illumina) were used for paired-end sequencing and index read sequencing, respectively.
Data analysis
Detailed descriptions of the data analysis methods, tools and information of used published data are provided in Additional file 1.
Real-time quantitative reverse transcription-PCR (qRT-PCR)
Total RNA was extracted from two patients (ALL 542 and AML 800) and four breast cancer cell lines BT-474, MCF-7, KPL-4 and SKBR3 with on column DNase treatment. The RNA was quantified using the Qubit fluorometer. For each sample, the RNA was divided into three fractions for total RNA and PA and RD processing. PA capture was carried out using Dynabeads mRNA Purification Kit (Thermo Fisher). RD was performed with Ribo-Zero Magnetic Gold Kit (Epicentre). The cDNA was synthesized using SuperScript III Reverse Transcriptase (Thermo Fisher). The qRT-PCR reactions were prepared using 10 ng of cDNA from each cell line or patient sample plus the iQ SYBR Green Super Mix (Bio-Rad, Hercules, CA, USA), and reactions run on the CFX96 Real Time System instrument (Bio-Rad). Normalized fold expression values were calculated by the ΔΔCt method using B2M, GAPDH, PGK1, and RPLP0 as reference genes and total RNA as control [21]. The primer sequences are listed in Additional file 2: Table S8.
PCR and Sanger sequencing
To validate the suspected fusion genes standard PCR was performed on cDNA from the ALL 542 and AML 800 samples. The cDNA was synthesized from total RNA using SuperScript III Reverse Transcriptase (Thermo Fisher). Primers were designed for ST3GAL1-NDAG1, MCM4-PRKDC, HBB-B2M, PQLC1-CTDP1, NCL-NR4A1 (Additional file 2: Table S8). The cDNA was amplified with Taq polymerase and using the T Professional thermocycler (Biometra, Göttingen, Germany). No template and GAPDH were included as negative and positive controls, respectively. PCR products were run on a 3% agarose gel, stained with GelRed Nucleic Acid Stain (Biotium, Fremont, CA, USA) and visualized on a standard UV trans illuminator. The DNA fragment for the HBB-B2M fusion gene was excised from the gel, cleaned using the NucleoSpin Gel and PCR Clean-up kit (Macherey-Nagel, Düren, Germany), and quantified using the Qubit dsDNA HS kit (Thermo Fisher). 4.5 ng of the fragment was used for Sanger sequencing using both forward and reverse primers of the HBB-B2M fusion gene and standard sequencing protocols.
Statistical analysis
Statistical analyses were performed with R version 3.3.1 (2016-06-21) and Prism software version 6.0 (GraphPad Software, San Diego, CA, USA). In the qRT-PCR analysis, two-tailed Student's T-test was used to analyze gene expression and P-values <0.05 were considered as statistically significant. Statistical dependence between two variables was calculated by Spearman's rank, Pearson's correlation analysis and hypergeometric distribution as implemented in R.
Overview of the study design
Reads and read mapping statistics
Expression landscape
We observed in total 30,205 features in the PA libraries as compared to 32,830 features detected in their matched RD counterparts using an RPKM (reads per kilobase per million) value of 0.125 as a threshold for minimum expression [22] (Additional file 3: Figure S2). While rather similar sets of features were captured, some differences were observed in specific transcript classes, such as processed pseudogene (7.82% to 7.03%), lincRNA (5.35% to 4.67%), and snRNA (6.99% to 6.54%) elements, as presented in a biodetection plot (Fig. 2b, Additional file 2: Table S1). Overall, the RD protocol detected 20.8 to 26.3% more of these features compared to PA. Antisense and miRNA features were also called at greater level by the RD protocol, while rRNA elements were enriched in the PA protocol. Notably, discrepancies in the calling of lincRNA, miRNA, and antisense genes resulted mainly from protocol differences, given the similar levels of these types of calls across the technical replicates (Fig. 2b). We also observed subtle but biologically intriguing variations in the detection of protein coding gene transcripts. For example, a total of 1380 protein-coding genes were called discordantly between the matched PA and RD libraries at RPKM threshold of 0.125 (Additional file 3: Figure S3), which is close to twice that observed between technical replicates (patient 668 and 1867; Additional file 3: Figure S4). Further analysis of discordantly identified protein coding transcripts revealed most of these to be attributable to RNA preparation (Additional file 2: Table S2). On one hand, 55 of a total of 71 histone genes (hypergeometric distribution p-value <0.05) were overlooked by the PA library protocol at RPKM value of 0.125. On the other hand, protein-coding genes relevant to cancer such as TGF-β1 mediating the activation of TGF-β/SMAD signaling pathway in ALL cells [23], BCL3, a proto-oncogene candidate associated with B-cell leukemia [24], and BRD4, which is associated with transcriptional deregulation in leukemia [25] were overlooked by the RD protocol at this threshold.
rRNA removal efficiency
To compare the rRNA depletion efficiency of the RD and PA protocols, the fraction of reads aligned to known human rRNA sequences in each library was quantified by aligning reads to the rRNA precursor sequences with the RNA-SeQC software [26]. Rather unexpectedly, the PA libraries exhibited higher rRNA mapping read rates than RD libraries (1.8% vs. 0.6%; Fig. 2d). However, the rRNA mapping rates varied highly (2.24% to 0.04% and 0.67% to 0.48%) even between technical replicate libraries.
Reproducibility of transcript abundances
Assessment of the concordance of protein-coding transcript abundances was made by measuring the correlation of RPKMs between different datasets (Additional file 3: Figure S6). We found a high level of concordance between the RPKMs of matched PA and RD samples (Spearman rho >0.95) and technical replicates (Spearman rho >0.98) in agreement with previously published results [18]. Since RPKM values between protocol-matched biological replicates (Spearman rho >0.91) and between AML and ALL samples (Spearman rho >0.88) were less correlated, patient heterogeneity appeared to represent the largest source of variation in these data. The hierarchical clustering of protein-coding transcripts with RPKM >4 and coefficient of variation >20 also corroborated the above findings and revealed that clustering was driven by disease type and patient rather than library type (Fig. 2e).
Accuracy in expression-based leukemia classification
Independent validation of RD and PA expression estimates
Fusion gene detection
To confirm some of the low confidence fusion genes, we tested the presence of five fusion partners with discordant fusion detections in two samples (AML 800 and ALL 542) using a standard PCR assay. PCR amplification of ST3GAL1-NDRG1 (5 spanning reads in ALL 542-PA), MCM4-PRKDC (3 spanning reads in ALL 542-RD), PQLC1-CTDP1 (6 spanning reads in AML 800-RD), NCL-NR4A1 (3 spanning reads in AML 800-PA) and HBB-B2M (8 spanning reads in ALL 542-RD) failed to detect the fusion gene, except HBB-B2M in ALL 542-RD. However, Sanger sequencing did not confirm the HBB-B2M fusion, which could be due to low abundance of the fusion in the sample or false positive prediction by the FusionCatcher tool (Additional file 2: Table S9).
Variant detection
In addition to the capture efficiency analysis, we performed variant effect annotation and filtering analysis on discovered variants. On average, 131,623 variants and 293 filtered variants were discovered in the process (Additional file 2: Table S5). Among these were several affecting one of the top 20 mutated genes for AML or ALL (Additional file 2: Table S6). For example, ALL 542-PA and ALL 542-RD had mutations in CREBBP (p.L161 fs) and ABL1 (p.T315I; present in 73 samples in COSMIC) genes, while AML 800-PA and AML 800-RD supported a mutation in the TP53 gene (p.R273C; present in 142 COSMIC samples). In addition to these three variants supported by both library preparation protocols, AML 800-PA revealed the presence of a mutation in the TET2 gene (p.V1949 fs). The library replicates detected mutations in DNMT3A, NRAS, TET2, BCOR, and CREBBP genes, indicating that variant discovery had high technical reproducibility and concordance.
Effect of library preparation on pathway enrichment analysis
With the unique ability to comprehensively characterize transcriptomes, RNA-seq has the potential to revolutionize clinical testing for a wide range of diseases. However, before the broader translation of RNA-seq into clinical practice, additional knowledge is needed to guide the selection of the library preparation protocol, as different alternatives can have a significant effect on downstream analysis and interpretation of RNA-seq outputs [12, 15, 18]. The performance of RNA-seq library preparation protocols has mostly been tested on non-clinical samples [16, 18–20] and the impact of protocols on clinical decision-making has not yet been addressed systematically. Some studies have even reported marked inconsistencies between RNA-seq data originating from differently processed libraries [19, 38], indicating that RNA-seq library preparation could also influence clinical decision-making.
To fill the gap in knowledge and assess the role of library preparation protocols on the detection of clinically important molecular characteristics, we applied two mainstream library preparation protocols to samples from leukemia patients and systematically compared their performance in a precision medicine setting. Although the small number of patient samples in our study is a limitation, especially for differential gene expression and pathway analysis, it absolutely mimics the clinical scenario and provides a fair equivalent to the current personalized medicine practices. Also, it is important to study the behavior of RNA-seq in a small-n setting to understand how RNA-seq performs in situations where only a few samples are available with no possibility of multiple replicates. Otherwise there is a risk that RNA-seq protocols are evaluated using metrics non-optimal for the goals of precision medicine and a wrong protocol is translated to standard clinical practice. Importantly, our study highlighted important differences between RNA-seq protocols, some of which were even clinically relevant, and indicated that that library preparation protocols have differing preferences for differential gene expression analysis, transcriptome characterization, fusion gene detection, and variant discovery.
Read and read mapping statistics demonstrated PA and RD libraries were largely comparable. All libraries were constructed from high quality RNA, had approximately similar insert sizes, and contained roughly the same amount of different types of reads. In line with recently published studies [12, 18], the PA protocol captured more transcripts emanating from exonic regions than the RD protocol. Given that these exon-mapping reads positively affect differential expression analysis [40], the PA protocol is the preferred method in understanding differential expression. In contrast, the higher intergenic or intronic mapped reads counts in the RD protocol can be advantageous in understanding pre-mRNA dynamics and identifying previously uncharacterized transcripts [17]. Moreover, the RD protocol captured 6.3% more non-coding RNAs compared to the PA protocol and depleted less non-coding RNAs in our qRT-PCR results suggesting its superiority at characterizing the non-coding RNA landscapes of the leukemia samples. In agreement with Sultan et al., [15] we found more rRNA mapping reads in the PA libraries than RD libraries, suggesting higher efficiency of the RD method to remove rRNAs compared to the PA method. This finding was further supported by expression estimation of rRNA genes in two patient samples by qRT-PCR. However, the rRNA mapping rates varied highly between technical replicate libraries, indicating that this step was greatly affected by experiment and preparation-dependent factors.
Regarding protein coding gene identification, the RD protocol performed a bit better and captured a wider repertoire. This method, for example, detected many non-polyadenylated protein-coding genes missed by the PA protocol, corroborating results from an earlier study [15]. Despite the better capture efficiency of the RD protocol, several known oncogenes were missed by this method. For example, TGF-β1, BCL3 and BRD4, which all have been linked with leukemia development [23–25] were overlooked by the RD protocol. This suggests that the PA protocol may suit better for characterization of leukemia transcriptomes and indicates that selection of the RNA-seq library protocol should be guided by the objectives of the study.
Protein coding transcript abundances were largely in agreement and a high level of concordance was found between the RPKMs of matched PA and RD samples. If extrapolated to other leukemia studies, our results indicate that biological features impact more on RNA-seq data reproducibility than library preparation. Moreover, disease heterogeneity rather than the protocol limits the comparison of RNA-seq data across leukemia studies. Nevertheless, the PA protocol quantified expression differences between AML and ALL better and provided constantly higher RPKM values for leukemia marker genes. This implies the suitability of the PA protocol for differential expression analysis is partly attributed to the higher number of exon-mapping reads in PA libraries, indicating that the read depth should be a key consideration in the adaption of RNA-seq to leukemia samples.
Validation by qRT-PCR also highlighted the suitability of the PA protocol for gene expression analysis. In particular, the PA protocol detected mRNAs efficiently and more accurately reproduced expression differences in clinical samples. Moreover, the PA protocol performed better in the analysis of breast cancer cell lines, emphasizing that the effect of the library preparation is consistent irrespective of the source of RNA material and type of cancer. In contrast, the RD protocol efficiently depleted rRNA molecules compared to the PA protocol and was better suited for the non-coding RNA detection. Overall, the qRT-PCR results suggested that PA better mimics total RNA when analyzing mRNA transcripts, which could be explained by the higher efficiency of the qRT-PCR reaction in PA libraries and presence of mature mRNAs in this RNA preparation.
Abnormal fusion genes caused by chromosomal rearrangement are important genomic events in leukemia and characterize a substantial population of the leukemia cases. For example, the BCR-ABL1 fusion gene is detected in 25–30% of young adult ALL cases [41] and is a clinical marker for treatment with targeted drugs. Markedly, both RNA-seq protocols successfully identified all known clinical diagnostic fusion genes BCR-ABL1, MLLT4-MLL and TCF3-PBX1 in the patient material with high numbers of supporting reads. This indicates that RNA-seq and fusion gene analysis can sensitively detect fusions despite some previous claims to the contrary [42]. In addition, many potentially false positive predictions were made and fusion genes were called rather discordantly even between protocol and technical replicates. However, most of the discordantly detected gene fusions were supported only by a small number of spanning read pairs, indicating that precision could be improved significantly using stricter filtering parameters. Results from PCR amplification of low confidence fusion genes supported by <10 reads also suggest that fusion genes supported by few spanning reads may be false positives and should be validated by other methods, if these fusions are of interest. However, validation methods such as PCR amplification followed by Sanger sequencing may not be sensitive to detect low expressed fusions.
Overall, this comparative study provides preliminary guidelines for the use of RNA-seq in a personalized medicine and other small-n setting, especially for hematological malignancies. In general, it showed that both PA and RD protocols produced consistent measures and were largely of similar usability. However, the PA protocol outperformed the RD protocol in more tests and it showed improved performance in gene expression analysis, classification of leukemia patients, quantification of leukemic marker genes, and variant analysis, which are all important for clinical sample assessment. Given that the study included only a limited number replicates, it would be beneficial to validate results using a larger cohort. Additionally, the effect of cDNA synthesis on library composition should be evaluated.
The authors would like to thank the patients who contributed samples to the study and personnel at the FIMM Technology Center including Pekka Ellonen, Aino Palva, Anne Vaittinen, Minna Suvela, and Henrikki Almusa for their technical assistance. Mika Kontro provided the clinical details of the patients.
The study was supported by funds from Finnish Funding Agency for Technology and Innovation (DNro 1488/31/09 and DNro 3137/31/13) and the Cancer Society of Finland (Syöpäjärjestöt). AK was supported by grants from the University of Helsinki Research Foundation and the Doctoral Programme in Biomedicine (DPBM), University of Helsinki.
Availability of data and materials
The data sets supporting the results of this article are included within the article and its additional files.
Authors' contributions
AK, MK and CAH designed the study. AK and MK performed the bioinformatics data analyses. AK and AP performed the cell culture work and qRT-PCR experiments. AK wrote the first draft. MK and CAH guided the analysis and supervised the work. AK, MK, PM, OK and CAH all contributed to the preparation of the manuscript. All authors approved the final version.
Ethics approval and consent to participate
The Helsinki University Hospital Ethics Committee approved the study and collection of samples (permit numbers 239/13/03/00/2010, 303/13/03/01/2011).
Consent for publication
Bone marrow (BM) aspirates from two AML and two ALL patients were collected after signed informed consent and with protocols in accordance with the Declaration of Helsinki.
Competing interests
CAH has received research funding from Celgene, Orion, Pfizer, and Novartis unrelated to this study. OK has research funding from Bayer and Roche unrelated to this study.
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text:
Set of 4 Volvo OEM Type R Performance Brake Calipers fits S60R V70R 04-07
- Brand: Volvo OEM
- Interchange #: CAL-ED041
- Volvo Part #: 30648978 // 30648979 // 30665550 // 30665551
- Stock #: CAL-ED041
- Availability: Out Of Stock
- * Please look at pics, condition and verify your fitment before purchasing, If you have any questions please contact us.
Not Available Online
Used Volvo OEM Set of Four Front and Rear Type R Brake Caliper Assemblies with attached brackets and brake lines.
Condition: Good condition with some wear and marks; See photos
Give this Genuine Volvo Used Part a Good Home!
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Stock Number: CAL-ED041
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Volvo Part#: 30648978 // 30648979 // 30665550 // 30665551
Color/Style: Gray w/ "R" Logo
Vehicle Fitment: Fits Volvo S60R V70R 2004-2007
Additional Notes: * Please verify fitment before purchasing.
We ship only within the Continental US. Most items ship out within 1 business day. All orders paid by Electronic Checks, parts will ship out after the payment has cleared through our bank. It will be helpful if you include your phone number and Volvo model & year to speed up delivery time and ensure accuracy. Domestic Shipping rates are based on per item. We ship all parts via UPS unless other is requested.
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If the item is defective or not as described, we will replace at no additional cost. All other returns will incur a 20% restocking fee.
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December 18, 1969
ACTOV - The U.S. Navy's Accelerated Turnover Program
America's Brown Water Navy in Vietnam is now the process of turning over most of its operating craft to the Vietnamese, training them in maintenance, logistics, and related fields, and serving as advisors to an emerging navy which has doubled in size in the space of one year.
At mid-1970, the U.S. Navy's in-country force will have relinquished most of its 500 patrol craft and gunboats and almost all naval operations to the Vietnamese Navy, through a program that will ensure continued success in the naval interdiction effort against communist infiltration and aggression.
That program is called ACTOV (Accelerated Turnover to the Vietnamese), and its task of training a well-drilled and competent force of 30,000 Vietnamese Navy Men, started last November, is in progress.
For the U.S. Navy, this means training the Vietnamese in riverine and coastal naval warfare, and turning over available surface craft and other assets. Before ACTOV ends, the Navy will train its Vietnamese counterparts in many fields from photography to PBR driving and will turn over craft as small as LCMs and as large as LSTs.
Vice Admiral Elmo R. Zumwalt, Commander U.S. Naval Forces, Vietnam, and Chief, Naval Advisory Group, and his staff have seen ACTOV throughits planning stages and successfully launched the concept in November 1968.
While U.S. Navy advisors, both officer and enlisted, worked in the field side by side with Vietnamese sailors, ACTOV experts at the Commander U.S. Naval Forces, Vietnam, headquarters in Saigon coordinate the transfer of Navy assets. ACTOV is represented at each major U.S. and Vietnamese Navy task force base to guarantee on-the-scene coordination.
It is the effort; however, of the individual boat crews and advisors who make ACTOV work for it is their job to see that the Vietnamese sailors are properly trained before the turnover of the craft can be completed.
Training is the key to the success of ACTOV and both navies operate several scholls for Vietnamese sailors.
Vietnamese Naval Training Centers (NTC) located at Saigon, Cam Ranh Bay, and Nha Tranh offer curricula similar to U.S. Navy specialty schools. Nha Tranh NTC offers "A" school or special training courses for electronics technicians, quartermasters, radiomen, engine men, electricians mates, and damage control men; Nha Tranh also houses the Vietnamese Naval Academy. Cam Ranh Bay NTC offers "A" school courses for commissary men, boatswain's mates, yeomen and gunner's mates. Saigon NTC offers lesser specialty short courses in its "C" school.
The U.S. Navy small Boat School is the first step in the training of Vietnamese sailors who will man the boats in the field. Using the audiolingual method of instruction, the students learn conversational English language patterns from American instructors. Then duplicate these sounds in a pronunciation drill, and begin to build sentences. The students then apply the English words to boat-oriented terms and equipment in each naval course.
The 12-week Small Boat school instruction also includes limited "laboratory" training aboard a River Patrol Boat (PBR) or a Swift boat (PCF) in the Saigon safe area.
Upon graduation, the Vietnamese sailor leaves for a three-month, on-the-job training tour under combat conditions with American sailors on river or coastline patrol.
After this, he becomes a regular crewmember aboard an American boat, replacing an U.S. Navy Man. The ACTOV process ends when Vietnamese sailors have completely replaced the Americans and the boat has been turned over to the Vietnamese Navy.
About half of the 500 boats which will eventually complete the Vietnamese Navy this summer have already been turned over. Thirteen "Swift" boats (PCFs), used mainly for coastal surveillance and patrol, were the latest assets to be transferred, and later turnovers in 1970 will be programmed as the ACTOV training schedule provides the qualified officers and men.
When all the boats in an American naval unit have been turned over, the Vietnamese also take charge of that unit's area of operations.
After all the boats are turned over, a big job still remains in the development of a successful support operation. Under the supervision of an ACTOV sub-unit called ACTOVLOG (for Logistics), many of the U.S. Navy river and coastal bases from the Cua Viet River near the Demilltarized Zone to the Ca Mau Peninsula, Vietnam's southern tip, will be gradually turned over.
Working concurrently with the turnover of craft but expected to take sometime longer, most of the bases will first be co-manned by American and Vietnamese sailors, then turned over completely when the Vietnamese can perform their own maintenance and logistics responsibilities.
The naval base at Beb Luc, some twenty miles southeast of Saigon, is now co-manned and will be turned over completely to the Vietnamese in the near future. A former Vietnamese Navy junk base at My Tho in the Mekong Delta was recently improved by U.S. Navy engineers and turned over in November 1969.
At the end of ACTOV's first year, the Vietnamese Navy has grown in size from 8,000 officers and men to more than 30,000. When the program has been completed the Vietnamese Navy will stand as the 14th largest naval force in the world.
To date, the ACTOV program has made possible the transfer of 242 boats and craft from the U.S. Navy to the Vietnamese Navy. The latest transfer of 13 Swift boats, the PCF, marks the half-way point in the turnover program. Subsequent turnovers in early 1970 will be programmed as the training schedule provides the qualified officers and men.
Most importantly, as the boats and craft are turned over, the Vietnamese Navy assumes responsibility for the areas of operations (AO's) previously under the control of U.S. Navy units. Initial accomplishments leave little doubt that the Vietnamese Navy is a viable and professional one.
The U.S. Navy through ACTOV is shaping the Vietnamese Navy into a formidable riverine and coastal force to eventually replace the Americans in Vietnam. The VNN will inherit a tactically successful interdiction operation on the rivers and an effective blockade along the coast. ACTOV is the pivotal program with the imporant function to see that this is done as smoothly as possible.
From the Cua Viet river in I Corps to the tip of the Ca Mau Peninsula in IV Corps, the "Great Green Fleet" carrying the colors of the Republic of Vietnam has come to do a job and in the end, just a few months hence, will force the "Brown Water Navy" into the annals of American history.


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UK Independence Party leadership hopeful Steven Woolfe MEP is conscious and recovering in hospital after an altercation with a fellow party MEP in Strasbourg.
In a statement, Mr Woolfe said:
"The CT scan has shown that there is no blood clot in the brain. At the moment I am feeling brighter, happier, and smiling as ever. As a precaution, I am being kept in overnight awaiting secondary tests to make sure everything is fine.
"I would like everyone to know that the parliamentary staff, the UKIP MEPs with me and hospital staff have been brilliant. Their care has been exceptional.
"I am sitting up, and said to be looking well. The only consequence at the moment is a bit of numbness on the left hand side of my face."
Several sources are reporting that the man who punched Mr Woolfe is Mike Hookem MEP, the party's Defence Spokesman, although this is unconfirmed. Police were not called to the incident and it is currently being treated as "a medical incident". It's thought that Mr Woolfe, the party's Immigration and Financial Affairs Spokesman, collapsed approximately half-an-hour after the incident took place, shortly after voting in the European Parliament chamber. Unconfirmed reports suggest that Mr Woolfe hit his head on a petal pole after being punched.
Mr Woolfe, a married father of one daughter, is 49 today and the early favourite to become the new leader of the party, after announcing his intention to stand yesterday. He previously stood in the last leadership contest but was blocked by the party's National Executive Committee. Diane James, who won the contest on September 16th, resigned after 18 days into the job on Tuesday night. Raheem Kassam, a former senior advisor to Nigel Farage and editor-in-chief of Breitbart London who is also running for leader, cancelled an appearance on BBC's Daily Politics out of respect for Mr Woolfe.
The news comes as an unexpected development in a saga enveloping the party. New Media Central Europe are hoping to bring more news about Mr Woolfe's condition and further detail on the incident as soon as we have it.
- Farage: Tory Brexiteers will "melt away" - November 6, 2018
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- UKIP's youth wing preparing for its biggest event yet #YIRevival conference announced for November 10th - August 17, 2018
- Guardian columnist calls on radio station to "dump" "far-right extremist" Farage - July 24, 2018
- "UNCONSCIONABLE" – Krishna Maharaj's 30-year battle for freedom NMC's Jack Oliver Smith speaks to human rights lawyer Clive Stafford-Smith about wrongly-imprisoned UK citizen - June 4, 2018


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Summer Seminar
July 24 – August 4, 2006
Theory from the Periphery:
Struggles for Social Justice
Seminar Leaders:
Michael Hames-Garcia, Binghamton University
Barbara and Carlisle Moore Distinguished Visiting Professor, University
of Oregon and Associate Professor of English, Binghamton University
Michael Hames-García grew up primarily in Oregon, where he
also attended college at Willamette University in Salem Oregon, receiving
a B.A. in English. He then moved to New York, earning a Ph.D. in
English from Cornell University and taking his first teaching position
at the State University of New York at Binghamton. He was granted
tenure and promoted to Associate Professor of English at Binghamton
in 2004, with joint titles in Comparative Literature and in Philosophy,
Interpretation, and Culture. He served as director of undergraduate
studies for the English Department from 2003-2005. He was also a
Hewlett Visiting Fellow at Stanford University's Research Institute
for Comparative Studies of Race and Ethnicity (RICSRE) in 2002-2003,
and is currently the Moore Distinguished Visiting Professor of English
at the University of Oregon.
Paula M. L. Moya
Associate Professor of English, Stanford University
A native of New Mexico, Paula Moya spent time in Texas (where she
earned a B.A. in English at the University of Houston) and New York
(where she earned a Ph.D. in English at Cornell University) before
coming to California in August 1996 to begin a career as an assistant
professor at Stanford. After being tenured in January 2002, Moya
served for three years as Director of the Undergraduate Program in
the Center for Comparative Studies in Race and Ethnicity (CCSRE)
and as Chair of the Comparative Studies in Race and Ethnicity (CSRE)
Seminar Description:
In this two-week, intensive summer seminar, we will be concerned
with analyzing, evaluating, and producing theory from the perspective
of minorities struggling for social justice. Some of the questions
we will ask include: How can theorists both develop theory praxically
and extrapolate it to larger contexts? How do we move from individual
experiences to larger social meanings? How might we analyze broader
social movements while attending to individual subjectivities?
How do our social contexts affect our personal choices? The readings
for the course will be drawn from the disciplines of sociology,
philosophy, literary theory, and psychology, and will feature such
thinkers as Linda Martín Alcoff, Manuel Castells, Daniel
Little, Maria Lugones, Walter Mignolo, Chandra Talpade Mohanty,
Satya Mohanty, Tobin Siebers, Julia Sudbury, Rosemarie Garland
Thomson, and Alison Wylie.
The seminar incorporates two workshops, one taught by the poet,
essayist, and activist Minnie Bruce Pratt (Syracuse University),
and the other jointly led by the social psychologists Hazel Markus, Dorothy Steele, and Claude Steele (all from Stanford University).
Seminar members will participate in the two-day colloquium organized
by the Future of Minority Studies Research Project on July 28-29.
Doctoral students who have completed at least two
years of their Ph.D. work and junior faculty in temporary or tenure-track
positions who are working on minority issues. Minority scholars and
those who are at HBCUs and other minority-serving institutions are
especially encouraged to apply. For the twelve scholars selected
to participate in the summer institute, subsidy will be available
to cover room, board, and (if needed) travel costs. FMS does not
charge tuition or fees. Application deadline: December 20, 2005.


text:
Our History
Creative Office Installations has been in operations since June of 1998.
Our Customers
We have serviced well over 300 customers and have business relationships with dealerships in the Sacramento region. We want to build long term relationships with all our customers.
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We are a full service commercial furniture installation company. Our installers have the training and experience to handle any size job while also meeting your budget and schedule. We provide turnkey and custom installations.
Our Mission
We pride ourselves on our customer service and completing the jobs on time. We focus on high quality craftsmanship and completing the job with minimal punch.
Our Team
We have a full staff ready to help and answer you requests.


text:
Copyright ©. Custom Home Pools | Privacy Policy
Your Custom Home Pool is designed to stay cleaner. Our efficient design
allows for fewer cycle times - saving energy! We use the most advanced engineering with a Dual Zone System. We use all 2" pressure pipe and schedule 80 2.5" Neverlube Valves. Then, we independently plumb all fixtures with their own dedicated pipe lines.We then pressure test all lines. The heart of our system is the energy-efficient Stealth Professional-Series 2220 Volt Pump, with a separate dedicated cleaner pump.


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Las Vegas, NV, 4/27/17-The World Players of Handball, WPH Outdoor, and 3WallBall are proud to present the 2017 WPH Outdoor 6 Stop #4 3WallBall Vegas Lte May 12-14, featuring the top men's, women's, and junior outdoor 3-Wall Big Ball handball players in the world. The WPH Outdoor 3WallBall 6 Stop #4 Vegas Lte is the fourth stop on the WPH Outdoor 6 season, as the top players continue their quests for ranking points and titles in "The Race to Vegas" and the WPH Outdoor 3WallBall Outdoor World Championships and Player's Cup Finale in September. The Vegas Lte event will be held at the Stratosphere 2-Court Employee Side Court Complex, 2000 S Las Vegas Blvd., Las Vegas, NV 89104.
The WPH Outdoor 3WallBall 6 Stop #4 Vegas Lte will feature 13-time WPH Outdoor singles champion and WPH Outdoor #1 Samzon Hernandez seeking to continue his dominance on the Outdoor 3-Wall courts. Hernandez will be joined in the Vegas Lte draw by New York's cross over handball superstar and Vegas Lte defending champion Timbo Gonzalez. Red Bull and Rollout sponsored Gonzalez earned his first WPH Outdoor 3-Wall singles title at the 2016 Vegas Lte, defeating then WPH Outdoor #1 Santos in one of the best singles finals in WPH Outdoor history.
Hernandez and Gonzalez will face a strong field at the Vegas Lte that includes WPH Outdoor stars Shorty Ruiz, Ricky Ruiz, Alfredo Morales, Josue Tellez, Sal "The Wiz" Duenas, and Chava Cordova. Each of the WPH Outdoor superstars has his sites set on the coveted Vegas Lte title and yellow jersey that accompanies any WPH Outdoor title.
Adding to the excitement at the 3WallBall Vegas Lte is the return of 9-time 4-Wall national pro singles champion David Chapman. The 13-year pro #1 4-Wall legend and former 3-Wall national champion will be making his first tournament start in more than three years and is eager to challenge himself against the best doubles teams on the WPH Outdoor tour. "I am looking forward to competing against the top Big Ball players in Las Vegas," stated Chapman. "I believe that Wil (Lemp) and I have a great chance in the doubles to win. With my front court skills and Wil Lemp's power, I believe we will match-up against any team."
When not competing, the WPH Outdoor stars will coach junior handball clinics and serve as mentors and role models for the next generation of players.
For information on the event, including how to enter and follow the draw, click here
For more information on the WPH Outdoor 3WallBall Vegas Lte, go here
WPH Outdoor Player's Cup: The World Player's of Handball's WPH Outdoor Player's Cup Series was formed in 2012 to showcase the game's best 3-Wall Big Ball Stars. Since 2012, the WPH Outdoor Player's Cup Series has hosted 33 events in California, Mexico, Utah, and Nevada, building the stage for hundreds of top big ball stars to compete against one another in professional, competitive and family-friendly events that promote sportsmanship and community spirit while inspiring the next generation of players. The WPH Outdoor 6 season features events in Southern California, Utah, and Nevada. All WPH Outdoor events host Junior WPH clinics with WPH Outdoor Elite players serving as mentors and coaches and junior tournament divisions starting at nine-and-under for boys and girls.
For more information on the WPH Outdoor Player's Cup Series, please contact World Players of Handball Executive Director David Vincent at email@example.com, World Players of Handball Development Director David Fink at email@example.com or WPH Outdoor Director Tom Flores at email@example.com. For more information on the World Players of Handball, please visit wphlive.tv


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I wanted to give a shout out to our favorite Friendly Local Game Store, DiceHouse Games in Fullerton.
We have not devoted much space to our gaming hobby here on OKOS, but once a month I run a White Wolf game under the banner of the local Dead Gamers Society of Orange County. At least that's what I do now. Honestly, I've been gaming since junior high, when I received one of the infamous "red box" sets of D&D from my equally geeky older brother.
But being a female gamer has it's challenges – one of the biggest being to find a game store where one can review books, talk shop with the owners, and sling dice with what is still a vastly male clientele, without feeling marginalized or like she's digging around a garage sale in her dorky little brothers basement. I've been to way too many shops that cover both.
As a college project, I imagined a game store with comfortable furniture, tastefully decorated and neatly stocked. A store with decent lighting. A store with clerks that didn't think that possessing a pair of breasts automatically made you unworthy of learning the finer points of putting together a killer Magic the Gathering deck, or who didn't just assume you must be shopping for your boyfriend.
Apparently someone else also had this vision, because not long after Dice House opened and it's been everything I've wanted in a game store since. Yes, I can get many books cheaper or sometimes even faster on the internet, but I like buying my gaming gear from people who know what good customer service means. I like store owners who take the time to get to know their regulars and will take it upon themselves to not only let us know when something we'd be interested in is available, but will set it aside for us. I like store owners that offer a safe, comfortable environment to play the games they sell, give you space to play games with other hobbyists without having to give out your home address (or making them deal with your cat/dog/crazy roommate), and who might even take the time to show you the ropes of a game your interested in learning.
There's something both very forward thinking and old fashioned about Dice House that should be encouraged, and I happily recommend them to any fellow gamers I run into that haven't already given their shop a go. Even if they don't have exactly what you're looking for, they are usually great about ordering it for you, or helping you find something else to tide you over until the next book release. Because that's how they roll.


text:
The Asian Development Bank (ABD) has warned that inflation will reach its highest level in a decade in most of Asia this year, threatening to reverse recent gains in productivity as well as creating fiscal strains for governments that provide massive subsidies for fuel and food.
The ADB haslowered its economic growth forecasts for the region as a global slowdown weighs on exports and expansions in China and India cool.
Bloomberg reports that Asia excluding Japan is predicted to expand 7.6 percent this year, less than a September estimate of 8.2 percent, the Manila-based institution said in a report today. The economies grew 8.7 percent in 2007, the fastest clip in almost two decades.
Central banks will pursue policies to quell inflation rather than spur economic growth, the ADB said, the second organization this week after the World Bank to warn of the threat of rising energy and food prices to the region. China will implement a ``tight'' monetary policy this year, the government's State Council said yesterday.
``The major risk lies not so much in softer growth but in rising commodity prices and accelerating inflation,'' the ADB said. ``Appropriate macroeconomic responses to accelerating inflation are likely to include tighter monetary policy and some exchange-rate appreciation.''
Crude oil rose to a record $111.80 a barrel last month, while prices of food staples including rice, wheat and soybeans and palm oil have surged amid increased demand and higher fuel and freight costs.
Wednesday, April 02, 2008
1 comment:
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