text:
I Know I can do it – a full potential that can only exist in our minds if it's not lived as the totality of who we are here in one single moment that it can be placed into physical/ practical living application- otherwise we then simply become great parrots.
Through reading Heaven's Journey To Life, I realized that this 'I Know' is no different to hope and having 'faith' in oneself which is a synonym of inaction and further waiting for something/ somehow being able to direct ourselves in the future, and in the meantime the 'I Know' remains as a point of security – as mentioned in the previous blogs, a false-confidence that serves no purpose other than holding a thoughtful-assertiveness without any physical results that prove it to be so in fact.
Continuing from:
- 235. Intelligence = Profitable Props for the Ego
- 236. Perfect Slave with Badges of Honor
- 237. Living IntelliSense as Life in Equality
- 238. I KNOW I Can
So, I've been debunking this self-belief of intelligence as just that: an accepted and allowed tag as 'who I am' that is no different to any other category we tend to reduce ourselves to. Therefore within this self-belief it is no different to holding the idea of a god in our minds having some form of power over others – same when I say 'I Know I can Do it' but not do it.
Memory within the 'I Know I Can' Character – Self Forgiveness and Self Corrective Statements
Event/ Memory: Being worried, stressed out, anxious about an exam when I was either in 2nd or 3rd grade primary school and my mother saw me all worried about it and told me 'You Know You Can, others that really can't should worry, but not you' within this creating a sense of security within the acknowledging of another that 'I can' because I simply didn't trust myself – and from this moment on, I would hear the same words in my mind the moment I would go into any form of stress about exams or any other academic point wherein my 'performance' was going to be measured. Within this, it became like a 'magical motto' that I would use to gain confidence but never really dissected what was it that was implied within this single 'You know you can' statement that became my 'I Know I Can.'
I forgive myself that I accepted and allowed myself to base a security of who I am according to Knowing what I am capable of based on previous results and experiences of getting a high-grade with certain ease wherein I would realize that stressing out was not required since I would always excel, which became a conceited way of existing as a knowledgeable character wherein I became of this knowledge-ego as myself as this certainty that became a self belief, creating a consequence of me really not being self-honest with myself because I stopped giving 'it all' that I could based on comparing myself to Others that would be in a much 'lesser position' and accordingly, measure my abilities/ capabilities as always remaining 'on top' but only through ranking systems wherein I stopped pushing myself further to actually develop skills or improve myself, but create a form of mediocrity wherein all that mattered is that I 'knew' how to do it,and would do it but that was it, there was no longer any push or drive because in my eyes and at the eyes of others, I was apparently already 'good enough' or 'intelligent' so
I forgive myself that I accepted and allowed myself to limit myself within a self belief of 'knowing' which eventually became only a fluffy ego self-belief without any form of actual work, substance behind it, becoming this façade as personality that people would identify myself with, without me precisely working on actually being/ becoming a 'better person' according to my standards then, but just keep myself 'on top' to remain within that position at the eyes of others, but knowing within myself that I wasn't really 'giving my all' and developing further skills and abilities, it only became an ego-driven effort to 'keep my spot' but not actually do it for myself for the purpose of actually expanding and learning more about myself, within this
I forgive myself that I accepted and allowed myself to become apathetic when it came to all things school, because within this ego of 'I know it all' everything became too repetitive within my mind, too useless, too dull and boring because of me believing that I was 'at the top' and there was nothing else to aspire to within this
I forgive myself that I accepted and allowed myself to spite my own predictability of becoming someone 'great' within the world system just like all the other kids with 'great grades' such as what I had back then, and because of deeming this 'excelling' within the system as something that I had to do but also resisted/ feared, I ended up opting for an 'unexpected choice' in life wherein I did all I could to dedicate myself to studies that had nothing to do with 'being intelligent' apparently, only spiting my own character and ending up trapped in my own web – so to speak – because of this choice being made out of spite and ended up spiting myself and getting caught in my own regret as another for of stagnation – which is unacceptable, because I was indeed the one that made the decisions and in no way are such decisions being considering what's best for all, but only spite the entirety of who I had accepted and allowed myself to be and become.
I forgive myself that I have accepted and allowed myself to believe that I had 'nothing else to do' or nothing else to be or become because I had it 'too easy' and as such, would get the same experience in any school or career because the problem was not the school or what I chose to study but my starting point and self experience within it and as such I forgive myself that I have accepted and allowed myself to blame the careers I chose, the people I chose to be with, the likes and preferences I developed and essentially the totality of my choices in life as something that 'didn't work,' without realizing that all of that was simply according to who I accepted and allowed myself to be as this knowing-character with no practical living considerations of what would actually be self supportive in self honesty, because I am well aware that I only sought my 'highest excitement' within my choices in life – therefore
I forgive myself that I accepted and allowed myself to make my choices in life as to what to study, who to establish relationships with, what to be 'inclined' to learn more about, the 'type of personality' that I became was all driven by/ through energy as the mind that sought a point of excitement within what I perceived as a dull life experience, within this not realizing that I deliberately sought to create a form of inner conflict to 'make my life more entertaining' as in having something to be sad about/ be regretful and essentially trapped in my own mind bubble of regrets, without realizing that no matter which choices I would have made, I was going to end up in the same position because there was no principle established in order to live in self honesty and within the consideration of self support, because that didn't exist in my frame of reference back then. Thus,
I forgive myself that I have accepted and allowed myself to try and blame others in my past and all the influences that I had throughout my life for having made the choices and decisions that I made, without realizing that it was only me that participated fully and fool-y within/ as them, playing the character for myself and others while holding a high expectation that was only sustained as a self-belief – not real as an actual physical movement of which one could only get the basics that would allow me to keep such position in place, but within me I Know that I didn't quite give it 'all' myself as an actual self-movement, which ensued apathy and dullness within myself toward anything having to do with being 'creative' or 'good' at something, not realizing that these energetic experiences were the consequential opposite outflow of first having chosen my career and studies based on an energetic high where no 1+1 was considered and I am fully responsible for that, and well aware of it, wherein
I forgive myself that I accepted and allowed myself to make decision in life based on energetic experiences of wanting to seek my 'feel good' point as a certain character/ personality that 'I knew' I could fulfill, without ever actually taking this decision and projecting it to see what are the practical points that I can employ myself in and being realistic about it according to how the world system works, which shows/ reveals that I wasn't in my 5 senses when I made the decision, but was existing as this 'fluffy' type of self-belief of me 'Knowing that I can make it' as this self-faith and self-belief of being 'capable of doing anything I can' but never really testing out and walking the practical physical considerations of what I would be able to work in and do within these careers and actually Do it as an informed decision – but, I didn't hence the consequences.
I forgive myself that I have accepted and allowed myself to hold my mother's words in a similar vein to the 'you know you an do it,' type of self-belief and hearing 'you are going to be great no matter what you do,' and in this believe that I could in fact excel and be the best 'no matter what I do' which is what positive thinking and talk lead to: creating a self-hope and self-faith of being potentially able to be the best wherever and whatever we do and become – which is not a 'bad' thing in itself, but how I lived those words as a form of false-certainty with no practical application.
I realize that within these words that became backchat, I became only a character that could live-out these self definitions within the law of the least effort wherein my results were compared toward others and within this, living up to simply 'maintaining' the character but not really being here as myself being self-directive in everything that I do, which is what we tie ourselves to within this world system: remaining as that specific character For others and neglecting an actual self-development to our fullest potential, which is not even encouraged within our current education system either no matter how many ranks and studies one may have, it is still within the confinement of a selective-preparation that one can do wherein the actual beingness of the person is – most of the times – not regarded, but only accumulating further knowledge and information as a form of 'betterment' that has never been based on actually supporting a human being to become a Living being in the consideration of equality as life or any other living principle that we can live by for the betterment of all – in essence, a blatant self-belief with no actual practical application.
I forgive myself that I accepted and allowed myself to elevate myself to my own expectations of being 'good' at whatever I decide to do, and take these words as an actual 'positive statement,' without realizing that if I would not in fact investigate in a practical physical matter what I would want to do, it would only became just another energetic-driven experience with no self awareness of the practical use of my decisions in life for an actual betterment of life. Within this
I forgive myself that I haven't accepted and allowed myself to realize that this ranking system in the school system more creates a sense of specialness/ uniqueness toward those with 'high grades' without really providing with an actual education and support toward actually being/ becoming something and someone that provides with an actual insight of the necessary points to change/ implement within this society for a common betterment, because none of this has been propagated or taught in schools, which then places into question the entire schooling system in itself, wherein if what's best for all is not placed as the common-goal within any field of studies, then: what are we actually supporting as 'education'? Who are we becoming within this education system within such terms and conditions of not really aiming at doing what's best for all? Is then a so-called intelligent person within this system that is not aiming at supporting life in equality an actual honorable definition of what Intelligence should be in this world'? No, it only becomes a synonym of being well trained to not ask questions, seek your permanent status within such 'higher ranks' in society, get well paid about it, become an example for others to follow which is what enables the system to be perpetrated generation after generation, no one questioning what these 'higher ranks' in society are actually based and founded upon.
Within this all, I forgive myself that I accepted and allowed myself to live out as an ego that believes can excel in 'everything I decide to do,' wherein it mostly became a sense of ingrained superiority against others as the primary self experience of 'knowing better/ knowing more' and creating this security based on what others believe me to be – only to then find myself obviously dismantling the entire self belief and realizing that if we look at what I actually did is nothing but just surfing on the ranks to maintain a mediocre superiority status for the sake of 'keeping my spot' as being intelligent/ responsible person, but the Who I was within such results was not really here as a self-directive being, actually doing it fully and whole heartedly, because I actually within this 'having faith' within myself, eventually 'lost faith' within myself and everything/ everyone, which became this constant apathetic self experience toward the world as the usual cycles of 'what's the use in this,' without realizing that this is the 'nitty gritty' point that I often hit as a continuous cycle once that 'I know how it will all end anyways' wherein I actually become the predicament of my own thinking processes, wherein we trap ourselves within our own loops of 'knowing how it all ends' and blindly driving ourselves to repeat the same experiences over and over again with no change – why? Because who I am within what I do is not fully self-honest as the realization that I had never in fact lived and that all I pretended to 'know' and be able to project an outcome of was only a way to 'give up' on myself before even starting, give up on any actual self-motivation because of seeing the world as just 'too fucked' to have any change be possible –
I forgive myself that I have accepted and allowed myself to still hold on to a belief deep inside that there is no way we can change humanity, which is the ingrained 'doomsday' type of self-experience that I became wherein I mostly lived up to others' expectations but not really placing all my beingness and effort and self-direction toward an actual living, doing all that I can to in fact develop myself further in a certain area or activity, just because of still holding on to this self-doom shadow as a constant presence of 'nothing that you do will work, will make a difference' which translates into an ingrained dullness and apathy within myself, giving up before even trying and just keeping a certain 'standard' but not really physically directing myself to for the first time motivate myself to live.
I forgive myself that I have accepted and allowed myself to stop challenging myself in life just because of believing that I have gotten to the 'peak' too fast and there was nothing else to 'attain' and within this, I forgive myself that I have accepted and allowed myself to equate life to an 'attainment' of sorts, believing that being alive was a synonym of getting 'high' in any form of rank of what 'success' is now measured within this world and reality, and within that realizing success for what it is, spiting it/ doing all I could to not become that, but only within an energetic-spite that then became as a hopelessness and uselessness wherein it did not matter how much I 'knew' it was 'useless' because it was never being placed within the consideration of what is best for all.
I forgive myself that I have accepted and allowed myself to instead of taking responsibility for our creation as this world system, create an experience about it, which is like complaining about our own fuckup as a form of victimization to not actually take self responsibility for it. Thus
When and as I see myself creating an Experience about our current world condition based on 'what I Know,' I stop and I breathe – I realize that in this I am only focusing on my own self experience and use it as an excuse to not move, instead of simply moving, doing and directing myself to face the consequences of which I am absolutely self responsible for in an equal manner, consider what's best for all and direct myself as it, physically – not thinking about it.
I realize that the perceived 'apathy toward living' is only an elitist self-experience that we are able to embody because we are secluded in this minute bubble of self-experience where the reality and the actual suffering in this world that is existent within everyone that have no access to money is simply disregarded, no considered – which proves how this apathy, hopelessness, helplessness and uselessness is only an egotistical experience wherein we are only looking at 'how we FEEL about life/ our reality' and create this entire self experience accordingly, instead of actually looking at the world physically, understand the flaws, understand how Nothing has worked in the past as a point of change and see where we have missed ourselves as living beings within it all.
I see, realize and understand that within this 'knowing' point, we diminish ourselves to our own self-beliefs as limitations that are and have never been based upon an actual consideration of Life, because Life is not a knowing but a living.
Within this, it is to currently realize that we have to walk the transition point from knowledge and information and all the characters we've become around this self-belief system of fake values, and individually walk our own self correction to establish a self-honest starting point within Anything that we are currently doing – because I realize that no matter what choices I would have made in life, I would have lead myself to the same 'uselessness' experience where nothing seems to work, nothing seems to make a difference because nothing of what is currently existing in this world is Meant to/ designed to/ created to make an actual difference in this world. Therefore
I commit myself to Live the realization that no matter where I am, what I do and the choices I make, nothing of what currently exists as the past and the old system that we are still living in is designed to 'work' and 'function' and be suitable for an actual birthing of life – because this IS precisely our task, our duty, our point of responsibility and within this
When and as I see myself ever again getting to the point of 'oh but nothing I do will change anything within the world/ others' without realizing that it is so, nothing will change and nothing will move if our starting point remains within the same old 'starting point' of this entire world system as it exist today, wherein nothing is veered toward an actual functional best for all outcome. Thus I realize that that is our work, that is our duty, that is our self responsibility that will not emerge 'by magic' but has to actually be conducted within self awareness of the required changes in the system in order to then be able to align ourselves to that which will create a substantial point of support for ourselves and others in this world.
I realize that there will be no quick fixes or results either, as such 'quick results/ fixes' exist only as a mind-interpretation of reality wherein no actual physical processes have been considered, nor any form of actual relationship that exists within ourselves toward each other and all the other living species, which then places into question how we have in fact never lived, because we have only ever equated ourselves to fulfill the same old standards within a system that has never really in fact functioned to support life.
Thus I commit myself to live the understanding and realization that a Knowing can only stand within the past of everything that we have been and become, and that nothing of this has ever in fact been living-conditions for each other – and that the motivation to then actually live won't come as a Knowing of anything that we've been in the past, but must be walked s a practical living daily application of letting go of everything that we Believe we know and instead, walk ourselves through a process of re-directing ourselves to consider physical reality outcomes and current systematic transitions wherein yes, we are in this world, we have to still present a knowledgeable act but! who we are within ourselves is an awareness of this being a single 'transition period' that we have to walk through, facing our failures and manifested consequences and within doing so, concomitantly paving the way for a new ay of living and existing, as I see and realize that the process of birthing ourselves as life won't have any 'precedents' within this system, it is an absolute self movement within the realization that it isn't preprogrammed, it won't come 'easy' or it won't be defined according to 'who I have been' in my past. It is entirely subject to my own self movement here in every moment of breath.
Thus, from the past we can take what is useful and what can be molded/ shaped or corrected in its starting point to be able to function within the Equality System as the Equal Money System, wherein people won't be regarded as 'more' for knowing what everyone will be equally capable of living/ doing – but instead, we will focus on supporting ourselves to develop practical skills that are readily useful to sustain ourselves in this world system as equals – no more hierarchical ranks in schooling systems of any kind.
I commit myself to stop existing in this self-hope of 'I know I can' but instead stop the past within myself by realizing that I don't require to have this self-positive-talk as assertiveness, but simply direct myself to do it, within the consideration of having to walk through the past in order to correct it here as myself.
I commit myself to stop regretting the choices I've made in life and believing that they lead me to failure, without realizing that all in this world is currently failing and that the only way that we can stand up for ourselves is walking through this failure, facing the consequences and concomitantly living the solution within our living application of doing, being, proposing what is Best for All.
I commit myself to live the realization that we have to walk through the past, give it an actual 'good riddance' as a self-corrective process to face what we have become, what we left behind and everything we 'Know' but didn't act upon and instead, give ourselves a start from scratch in order to actually build/ create what we are willing to stand equal and one to as our self-awareness creation, to no longer have to create a 'negative' or 'positive' experience about ourselves and 'who we are' within this world, but instead simply focus on aligning ourselves to be and become equal contributors to a best for all outcome in this physical reality – no more and no less, using the knowledge and information that can provide us with actual insights and data that we can consider in order to establish a world system based in Equality, the same process that we can apply for ourselves to not maximize our capabilities only at a thought level, but rather physically supporting ourselves and each other to ground such potentials toward an actual doing that ensues a result that benefits the whole in equality.
To be continued…
For further support:
Desteni Lite Process – Free Online Course
The following illustration depicts this point, it is only going back to the ground wherein we can rebirth ourselves from scratch. It is then quite obvious how we could not be able to make ourselves 'succeed' from the starting point of everything we have believed ourselves to be. This is our equal point of starting from scratch, and this is how no one can possibly be 'more' than others or 'more advanced' as nothing in this world can possibly provide such advancement without the illusion of progress being tainted by self-abuse.
Time to live in common sense and realize that every breath is our equal-starting point as physical beings that can direct ourselves to live the words we write/ speak in common sense – this will prevent any form of perceived 'failure' in this world system, as we have all in fact already failed to live as equals – hence, the willingness to walk this self-corrective process: there is no way out of this, we have to face our creation.
Good riddance to any form of laureate past since everything we have done and become in this world has been based and founded upon abuse, there is no way to keep any form of honor within what we've become as that would be ego and self separation.
HOPE – the Metaphysical Carrot (Part 3): DAY 239
Day 239: Assumption as God – ADC – Part 86
This blog is part of the After Death Communication Series


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Maryland Injury Guys
Maryland Injury Guys Fulton Personal Injury Attorneys
Maryland Injury Guys Fulton, MD was founded to help the community who were injured in car accidents, work accidents, dog bites, bicycle accidents, and those who suffered a medical malpractice, get justice for the negligence caused by others. Our local attorneys do this through personal injury lawsuits, and workers' compensation claims and appeals. Call our local Fulton law office for a free consultation. Our office serves Fulton, MD. North Laurel. MD. Laurel. MD. Savage. MD. Annapolis Junction. MD. Maryland City. MD. Burtonsville. MD. Spencerville. MD. Cloverly. MD. South Laurel. MD. Maple Lawn. MD. Russett. MD
Business Name:
Maryland Injury Guys
11820 W Market Pl Suite K-1Fulton Maryland 20759United States
(410) 208-7309
About Fulton
Fulton is a census-designated place located in southern Howard County, Maryland, United States. As of the 2010 census it had a population of 2,049. == History == Indigenous peoples, likely Piscataway, used the land now known as Fulton for hunting and farming. The land's first European survey was by Thomas Browne, known as the "Patuxent Ranger", in 1700. In the mid-1700s Richard Snowden, the Quaker grandson of one of Maryland's first iron ore producers, purchased tracts of land up the Patuxent River valley.
Neighborhoods in Fulton, Maryland
Allview Estates, Arbor Terrace Fulton, Ashton Pond, Bryant Woods, Clarksville Ridge, Crestwood Neighborhood, Dickinson, Donleigh, Dorsey's Search, English Manor Neighborhood Park, Fulton Elementary School, Glenmont, Glenmont, Glenshire Towne, Hallowell
Our Newest Profiles

subset: commoncrawl


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Our President
Adam Weinberg
Adam Weinberg joined Denison University as its 20th president on July 1, 2013. He previously served as president and CEO of World Learning, one of the premier international education, exchange and development organizations. World Learning works with young people from more than 140 countries, helping them develop the ability to address critical global issues.
Previously, he was vice president and dean of the college at Colgate University, where he was a member of the sociology department for more than a decade. At Colgate, he gained national prominence for his work on increasing the level of civic engagement at colleges and universities. He also founded a number of organizations, including the Center for Outreach, Volunteerism, and Education (COVE) and the Partnership for Community Development.
Weinberg brings to Denison a breadth of experience with the liberal arts and global education. He was drawn to Denison by its innovative faculty and motivated students; its momentum on the landscape of higher education; and its academic rigor, scholarly research, civic engagement and steadfast commitment to the liberal arts.
"The world desperately needs the kind of graduates that Denison produces," he says. "The values, skills, habits and knowledge of the liberal arts are critical to the next generation of political leaders, social entrepreneurs and private sector leaders."
A native of Texas, Dr. Weinberg's passion for ice hockey took him to New England, where he attended Deerfield Academy and Bowdoin College, graduating magna cum laude. He studied at Cambridge University before earning his master's and doctoral degrees in sociology from Northwestern University. He has co-authored two books, Urban Recycling and the Search for Sustainable Development and Local Environmental Struggles, and currently is writing a book on the relationship between higher education and civil society. He also has published articles in The Washington Quarterly, The Chronicle of Higher Education, Inside Higher Education, Peer Review and a range of academic journals.
He is a member of the Council on Foreign Relations and of the Higher Education Working Group on Global Issues, and has served on a variety of national and local boards including the Alliance for International Education and Cultural Exchange, InterAction and Vermont Campus Compact. When he arrived at Denison, Weinberg joined the boards of Ohio Campus Compact, The Works and Our Futures of Licking County.
Dr. Weinberg resides in the President's House at Denison, with his wife, Anne, their three children and a new puppy.
Visit the President's Office for a comprehensive view of presidential activities.


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The Motorola Atrix 2 may be one of the most underrated smartphones on the market. It combines a lot of high-end features with a great price tag ($99 base price) and has the unique value proposition of Motorola Webtop for docking as a laptop or desktop. That's why it's our pick for Product of the Week in our inaugural episode of this new video show.
Learn which types of users can benefit from the Atrix 2, the pros and cons of the product, what it means for business and IT, and the information you should know about it the Atrix 2 that you won't find in other reviews. Watch the first episode of this new weekly series. You can also find a quick summary below.
Who's it for?
If you want to do everything possible with an Android smartphone, then the Motorola Atrix 2 is a strong candidate because of its mix of multimedia capabilities and business readiness. Its Webtop software lets you dock it into a laptop or desktop experience and surf the web with Firefox or connect to the corporate network using the Citrix client. The Lapdock can replace a tablet or netbook for some highly mobile users. This phone is also for power users who are looking for a bargin, since it starts at US$99.
- Loaded with high-end features
- Great price, starting at US$99
- Thin but sturdy hardware with a nice feel
- Motorola's Webtop docking experience
- HSPA+ capability (but don't call it "4G")
- Webtop software still has performance issues
- Its 4G is really "faux G"
- Uninstallable AT&T apps (a.k.a. "crapware")
Product of the week: Our approach
If you're looking for a quick two-minute summary of a new consumer product, then this isn't the series for you (check out CNET Reviews). This show will do a deep dive on a product and look at where and how it makes sense to use it for business. The format is highly conversational and each episode will be about 15-20 minutes. You'll occasionally see an episode the week a product comes out — when we get early access to a product — but more often than not, the Product of the Week will be a product that has been on the market for at least a couple weeks so that we can get our hands on it and really use it before making a determination about it.
Since we're just getting started, we'd like your feedback on this format. Does it work? What else would you like to see? What are you looking for in a thorough product review?


text:
- Research
- Open Access
- Published:
Determinants of selenium status in healthy adults
Nutrition Journal volume 10, Article number: 75 (2011)
11k Accesses
107 Citations
9 Altmetric
Selenium (Se) status in non-deficient subjects is typically assessed by the Se contents of plasma/serum. That pool comprises two functional, specific selenoprotein components and at least one non-functional, non-specific components which respond differently to changes in Se intake. A more informative means of characterizing Se status in non-deficient individuals is needed.
Multiple biomarkers of Se status (plasma Se, serum selenoprotein P [SEPP1], plasma glutathione peroxidase activity [GPX3], buccal cell Se, urinary Se) were evaluated in relation to selenoprotein genotypes (GPX1, GPX3, SEPP1, SEP15), dietary Se intake, and parameters of single-carbon metabolism in a cohort of healthy, non-Se-deficient men (n = 106) and women (n = 155).
Plasma Se concentration was 142.0 ± 23.5 ng/ml, with GPX3 and serum-derived SEPP1 calculated to comprise 20% and 34%, respectively, of that total. The balance, comprised of non-specific components, accounted for virtually all of the interindividual variation in total plasma Se. Buccal cell Se was associated with age and plasma homocysteine (hCys), but not plasma Se. SEPP1 showed a quadratic relationship with body mass index, peaking at BMI 25-30. Urinary Se was greater in women than men, and was associated with metabolic body weight (kg0.75), plasma folate, vitamin B12 and hCys (negatively). One GPX1 genotype (679T/T) was associated with significantly lower plasma Se levels than other allelic variants. Selenium intake, estimated from food frequency questionnaires, did not predict Se status as indicated by any biomarker. These results show that genotype, methyl-group status and BMI contribute to variation in Se biomarkers in Se-adequate individuals.
That selenium (Se) plays roles in cancer prevention has been demonstrated in numerous studies with a variety of animal and cellular models [1–4] and in several clinical trials [5–7]. Yet, the largest study, the Selenium and Vitamin E Cancer Prevention Trial (SELECT), failed to detect cancer risk reduction with Se-supplementation [8], indicating that Se-supplementation may not benefit all individuals.
Rayman et al [9] pointed out that the results of SELECT are consistent with those of the Nutritional Prevention of Cancer (NPC) Trial [7, 10]. In NPC, reduced prostate cancer risk was observed mostly among subjects with baseline plasma Se concentrations in the lowest tertile of the cohort, < 106 ng/ml [10]; no risk reduction was noted for subjects with plasma Se > 123 ng/ml, concentrations comparable to those of the subjects in SELECT, which averaged 136.5 ng/ml at baseline [8]. Therefore, it is possible that Se-supplementation may yield anti-cancer benefits in individuals who are below some threshold of Se status without being deficient in the nutrient.
Assessment of Se status is difficult in individuals who are not deficient, such as those of NPC and SELECT. It is well established that Se-deficient individuals show subnormal levels of several Se biomarkers, including some with functional significance, such as the selenoproteins, and others that indicate the amounts of Se in the body, such as the Se contents of tissues and body fluids. Non-deficient individuals, however, have maximal selenoenzyme expression [11–13], rendering those parameters non-informative regarding changes in Se intake. Thus, the total Se content of plasma has become the default biomarker of Se status in non-deficient cohorts.
Plasma Se, while easily measured, is not a single entity. It has several components, which, with our current knowledge, are currently defined as: two selenoproteins (selenoprotein P [SEPP1] and the extracellular GPX3), which specifically contain selenocysteine (SeCys) [14, 15]; Se incorporated non-specifically as SeMet in lieu of methionine in albumin and other proteins [16, 17]; and a small amount of non-protein bound Se [18, 19]. These selenoproteins are fully expressed in Se-adequate individuals; they comprised 22% (SEPP1) and 9% (GPX3) of plasma Se in an individual fed SeMet [19]. In contrast, the non-specific incorporation of Se into plasma proteins appears to be regulated only by SeMet supply for which reason this component would be expected to increase in response to supplementation with SeMet [19], as in SELECT [7], or a source of SeMet such as Se-yeast, as in NPC [8].
We determined multiple biomarkers of Se status in the cohort of healthy Americans in order to elucidate the relationships among those biomarkers and biological, metabolic and genetic factors relevant to Se metabolism. Herein we report the results of an extensive assessment of Se status in that cohort.
This study involved healthy men and women living in vicinity of Grand Forks, ND, who volunteered and met the following eligibility criteria: over 18 yrs., no chronic liver or kidney diseases (based on blood chemistries and urine analyses), nonsmokers, not having used supplements providing > 50 ug Se/day in the past 6 months (to accommodate users of most OTC multivitamin/mineral supplements), not currently using medications that adversely affect liver or kidney function, body mass index (BMI) ≤ 40 kg/m2. A total of 356 volunteers were screened. Of 282 who met the eligibility criteria, 261 subjects (106 men, 155 women) were enrolled in the study; selection and randomization of subjects is shown in figure 1 and subject characteristics are indicated in table 1. Each volunteer was given a cash honorarium pro-rated for the duration of his/her participation in the study.
Oversight was provided by the University of North Dakota Human Subjects Committee which reviewed and approved the protocol. The purposes and procedures of the study were explained to the volunteers verbally and in writing, and written informed consent was obtained from each volunteer.
Body weight was measured using an electronic scale. Height was measured using a wall-mounted stadiometer.
Assessment of Dietary Intake
Dietary intake over the three previous months was assessed by a single, self-administered food frequency questionnaire (FFQ) patterned after the Harvard Service Food Frequency Questionnaire format [20]. This FFQ includes 78 food items without serving sizes indicated (natural portion implied: e.g., one cup of milk, one slice of bread). All food items on the FFQ were matched to food codes from the USDA Nutrient Database for Standard Reference [21] or USDA Food and Nutrient Database for Dietary Studies [22]. For each food item, subjects designated their average consumption by marking one of nine frequency categories ranging from "zero per month" to "six or more times per day". The frequency chosen for each food item was converted to daily intake, e.g., a response of "1-3 per month" was converted to 0.07 servings per day (two servings per month). Total energy, protein, carbohydrate and fat intake were calculated based on nutrient data for the food codes matched to the FFQ food items. Selenium intake cannot be calculated as selenium content of foods varies widely, dependent on crop location. We therefore utilized the Se core foods list developed by Schubert et al [23] to provide suggested food intake patterns. Foods were also sorted into groups using information from the MyPyramid Equivalents Database for USDA Survey Food Codes [24] as well as Friday and Bowman [25].
Sample Collection and Preparation
Blood was collected by venipuncture in duplicate 7 ml samples into heparinized, EDTA-treated and non-treated glass tubes. Aliquots of whole blood were subjected to low-speed centrifugation to prepare erythrocyte, buffy coat, plasma and serum fractions. Urine (24 hr samples) was collected in sterile polycarbonate bottles. These specimens were held at 4°C pending the completion of screening analyses; excess portions were held at -80°C. Buccal cells were collected using a sterile toothbrush according to Paetau, et al. [26]; cells were lysed in distilled water and lysates were held at -80°C for analysis.
Clinical Biochemical Analyses
Blood cells were counted using an automated system (CellDyne 3500, Abbott Laboratories, Abbott Park, IL). The activities of aspartate aminotransferase (E.C. 188.8.131.52) and alanine aminotransferase (E.C. 184.108.40.206) were determined in serum using kits (AST2 and ALT2, respectively, JAS Diagnostics, Inc., Miami, FL) and an automated chemistry analyzer (Cobas Mira, Roche Diagnostic Systems, Inc., Sommerville, NJ). The same analyzer was used to determine blood urea nitrogen (BUN) in serum, glucose and protein in plasma, and creatinine in urine (#47381, #0458976160, #44903 and #47003, respectively, Roche Diagnostic Systems, Inc., Sommerville, NJ). Thyroid-stimulating hormone (TSH), non-protein bound thyroxine (T4) and triiodothyronine (T3) were measured in serum using an automated, solid-phase, two-site chemiluminescent immunometric assays (LKTS1, LKF41 and LKF31, respectively, Immulite 1000 System, Diagnostic Products Corp., Los Angeles, CA).
Biomarkers of Se Status
Selenium status was assessed on the basis of the activity of GPX and the amount of SEPP1 in serum, and the amounts of Se in plasma, buccal cells and urine. The activity of GPX3 (E.C. 184.108.40.206) was determined in plasma by the method of Paglia and Valentine [27] as modified by Lawrence and Burk [28]. Due to its affinity for heparin, SEPP1 was measured in serum by an enzyme-linked immunoassay [29]. Selenium was determined in plasma, buccal cells and urine by automated electrothermal atomic absorption spectrophotometry using a reduced palladium matrix modifier and an instrument equipped with L'Vov platforms [7]. Certified Standards were used (Alfa Aesar [Ward Hill, MA, USA], Perkin Elmer [Waltham, MA, USA] and CPI [Santa Rosa, CA, USA]) to prepare a calibration set daily with each batch. Continuing calibration certification and a continuing calibration blank were included at 10% frequency and at the beginning and end of the daily batch. An initial calibration verification from an alternate supplier was validated the calibration set at the beginning and end of each analytical series. Matrix effects for plasma and urine were evaluated using quantitative plasma and urine standards (NIST [Gaithersburg, MD, USA], Seronorm [Billingstad, Norway] and Utak [Munich, Germany]) to validate the percentage recovery of the analyte in these sample matrices. As there is no commercially available quantitative standard for Se in buccal cells, matrix effects of buccal cell preparations were accounted for in the analysis by performing spike recoveries using certified calibration standards added directly to one of the samples.
Other Biochemical Analyses
Folic Acid, B12 and homocysteine (hCys) were measured in serum by automated solid-phase, competitive chemiluminescent enzyme immunoassays (LKF01, LKVB1 and LKH01, respectively, Immulite 1000 System, Diagnostic Products Corp., Los Angeles, CA). Vitamin 8-Hydroxy-2'-deoxyguanosine was measured in urine using a competitive enzyme-linked immunosorbent assay (#21026, Bioxytech, Oxis Health Products, Inc., Foster City, CA).
The genotypes of subjects were determined for genes for several selenoenzymes (two glutathione peroxidases, GPX1, GPX4; the transporter selenoprotein P, SEPP1; and intracellular SEP15). Genomic DNA was extracted from blood samples using a DNA isolation kit (Qiagen, Valencia, CA). DNA was used as a template for PCR amplification by modifications of previous reported procedures [30–33]. Negative controls were included. Unless otherwise noted, PCR conditions were: 95°C - 3 min; 30 cycles at 95°C - 30 s; 55°C - 60 s; 72°C - 90 s; 72°C - 10 min. Amplified DNA was digested with the appropriate restriction enzymes, and digestion products were separated electrophoretically in 3% agarose unless otherwise noted.
SEP15; rs5845 and rs5859
PCR primers [30] 5'-CAGACTTGCGGTTAATTATG-3'and 5'-GCCAAGTATGTATCTGATCC-3' were used to generate a 413-bp amplification product of rs5845 and rs5859. Amplified DNA was digested with Dra I restriction enzyme at position 811, or Bfa I restriction enzyme at position 1125.
GPX4; rs713041
PCR primers [31] 5'-GACCTGCCCCACTATTTCTA-3' and 5'-GTCTGTTTATTCCCACAAGG-3' were used to generate 221-bp amplification product of rs713041. PCR conditions were as follows: 94°C - 2 m; 30 cycles at 94°C - 30 s; 57.5°C - 30 s; 72°C - 1 min; 72°C - 7 m. Amplified DNA was digested with restriction enzyme StyI.
GPX1; rs1050450
PCR primers [32] 5'-TGTGCCCCTACGCAGGTA-3' and 5'-CCAAATGACAATGACACAGG-3' were used to generate a 337-bp amplification product of rs1050450. Amplified DNA was digested with the Apa I restriction enzyme to identify C/T polymorphism at allele position 679, corresponding to amino acid position 198 of the human GPX1 protein.
SEPP1; rs3877899
PCR primers [33] 5'-CACGCATTATTCCTATCTCTATAAGCTTG-3' and 5'-GGAAATGAAATTGTGTCTAGACTAAATTGG-3' were used to generate a 722-bp amplification product of rs3877899. To determine a G/A variant at position 24731 in SEPP1 mRNA and a second G/A variant (rs7579) at position 25191 of the reference mRNA sequence in the 3'UTR of mRNA, PCR products were sent for sequencing (Cogenics, Newton, MA) with sequencing primers 5'-CACGCATTATTC-CTATCTCTATAAGCTTG-3', 5'-TCACCTGACA-GTGTAAAGAAAACCTC-3'.
Statistical Analyses
All statistical analyses (Pearson and Spearman correlations, regression analysis, ANOVA, non-parametric discriminate analysis, Tukey contrast and orthogonal contrast) were performed using SAS Version 9.1.3 (SAS Institute, Inc., Cary NC). Data for buccal Se, urine Se, SEPP1, folate, hCys and vitamin B12 were highly skewed and were logarithmically transformed so that their distributions would more closely approximate a normal distribution. For these variables, we report the geometric mean with a 1 SD interval. All other data are expressed as mean ± SD Pearson correlations and linear regressions were computed between biomarkers of selenium status and demographic variables, Se intake, and measured biochemical parameters. Because of the large variability observed in the intakes of the food groups, Spearman correlations were used to assess the relationship between plasma Se and intakes of various food groups. A discriminate analysis was performed to determine whether estimated Se Intake could predict the relative magnitude of plasma Se using the first and fourth quartiles of the latter. A nonparametric method was used because the intake values were not normally distributed. For the SEPP1 and BMI relationship, further analysis was performed by use of both Tukey contrast and orthogonal contrast to validate the correlation.
Estimation of Dietary Selenium Intake
This cohort was of relatively high Se status, as indicated by plasma Se level (142.0 ± 23.5 ng/ml) (figure 2) other biomarkers of Se status (Table 1) and estimated daily Se intake of 109.1 ± 43.6 μg/d. The major dietary sources of Se were whole wheat bread/rolls, eggs, and spaghetti/other pasta with sauce, which accounted for 21.8% of estimated intake. The estimated Se intakes of men (122 ± 51 μg/d) were highly significantly (p < 0.001) greater than those of women (101 ± 35 μg/d). A core group of 22 foods provided 80% of the estimated amounts of Se consumed by this cohort; 9 of those foods provided half of estimated daily Se intakes. The rankings of these foods as sources of Se were similar for men and women.
Se Distribution in Blood
The major components of plasma Se were estimated from the total plasma Se and the measured plasma GPX3 activities and serum SEPP1 amounts using the following assumptions; for the amount of SeCys, GPX3 enzyme rate constant of 2.8 × 104 nmol/min/mg, molecular weight 92 kD, and (stoichiometry of 4 g-atoms Se per mole as SeCYS [34]; for SEPP1 (glycosylated), average molecular weight 60 kD and stoichiometry of 9.9 g-atoms Se per mole as SeCYS [35]. An assumption inherent in the selection of this blood fraction for SEPP1 is that insignificant amounts of SEPP1 protein are removed during the clotting process. We employ this common assumption, with the further caveat that SEPP1 loss by clotting is assumed to be less than the variance of SEPP1 found in the studied population (Coefficient of Variation ~27%).
The difference between the total measured Se and the amounts of Se corresponding to the activity of GPX3 and measured amount of SEPP1 was taken as the amount of Se incorporated non-specifically into plasma proteins, presumed to be predominately SeMet. By these estimates, GPX3 and SEPP1 comprised approximately 20% and 34%, respectively of Se, while 47% of Se was present as non-specific components (Table 2). Neither the activity of GPX3, the amount of SEPP1 in serum, nor the amounts of Se in buccal cells or urine were significantly related to total plasma Se concentration. However, the non-specific component of plasma Se was positively associated (r = 0.87, p < 0.0001) with plasma Se (figure 3). SEPP1 concentration was associated with plasma concentration of hCys, a marker of methylation status (r = 0.13, P < 0.05) on an exponential basis.
Serum SEPP1, but not GPX3 activity or total plasma Se, showed a significant quadratic relationship with body mass index (BMI) (r2 = 0.054, P < 0.002), being lower in individuals at the low and high ends of the BMI range (figure 4). In addition to the regression analysis shown in figure 4, ANOVA, Tukey contrast and orthogonal contrast tests were performed. There were only 2 subjects with BMI < 18; these were included in the BMI < 25 group. The overall ANOVA was highly significant, p = 0.0012. By Tukey contrast tests comparing the means of the 3 groups, the mean of the lowest BMI group was highly significantly less than the mean of the overweight group, p = 0.0008. An orthogonal contrast test showed a significant quadratic trend in the means of the 3 groups, p = 0.003. The back-transformed SEPP1 means (± 1 S.D.) are as follows: BMI < 25: 3.17 (3.07-3.26); BMI 25-30: 3.69 (3.59-3.80); BMI > 30: 3.46 (3.36-3.58). Figure 4 makes evident the presence of two outlier subjects. With all values included in the quadratic regression of SEPP1 vs BMI, R2 = 0.054, p = 0.0016. Excluding the two high values improved the fit slightly: R2 = 0.0596, p = 0.0009.
Determinants of Se Status
Plasma Se level was not significantly associated with Se intake estimated by FFQ (figure 5); however, it was significantly associated with numbers of servings of foods in four food groups: fish (r = 0.14, P < 0.022), fish subcategory "other" (including fish other than tuna; r = 0.15, P < 0.014); dairy (r = 0.13, P < 0.036), and cured meat (including sausage and luncheon meats; r = 0.15, P < 0.019). A discriminate analysis showed that food Se intake correctly predicted the plasma Se quartile for about 73% of the individuals. The foods selected by the analysis are shown in Table 3. Plasma Se level was not significantly different between men and women, or between users and non-users of nutritional supplements. We assumed that the maximum amount of Se consumed from supplements (<50 ug/day) would fall within normal day-to-day interindividual variations in Se-intake. That assumption is borne out by our finding of no significant differences in the plasma Se levels between supplement users and non-users for plasma Se. Plasma Se was not significantly associated with age, metabolic body weight (kg0.75), or serum concentration of folate, vitamin B12 or hCys.
Buccal cell Se was not significantly different for men vs. women; however, that of users of nutritional supplements (9.22 [5.89, 14.45] 4 ng/mg protein, n = 81) was significantly greater (p < 0.02) than that of non-users (8.03 [5.50, 11.73]4 ng/mg protein, n = 179). Buccal cell Se was significantly associated with age (r = 0.14, P < 0.02) and with serum hCys (r = 0.24. P < 0.0001) on exponential bases. It was not, however, associated with BMI, metabolic body size, or serum concentrations of folate or vitamin B12.
The urinary Se level of women (geometric mean [-1S.D., +1 S.D.]; 57.7 [42.3, 78.6] ng/mg creatinine) was significantly greater (p < 0.003) than that of men (51.8 [39.8, 67.5]3 ng/mg creatinine); that of users of nutritional supplements (geometric mean [-1S.D., +1 S.D.]; 59.3 [44.0, 80.0] ng/mg creatinine, n = 80) was significantly greater (p < 0.01) than that of non-users (53.5 [40.0, 71.5] 3 ng/mg creatinine, n = 180). Urinary Se was not significantly associated with age or BMI; but it was significantly associated with metabolic body weight (kg0.75) (r = -0.14, p < 0.024) and with serum levels of folate (r = 0.17, P < 0.006) and vitamin B12 (r = 0.34, P < 0.0001) and negatively with serum hCys (r = -0.16, P < 0.009) on exponential bases.
This cohort showed heterogeneity with respect to the genotypes for each of the selenoproteins determined, three of which were significantly related to a biomarker of Se status (Table 4). Individuals with the GPX1 679 T/T genotype (rs1050450) constituted 11% of the cohort and had significantly lower (by 7%) plasma Se levels than those with the most prevalent (47%) GPX1 679 C/T genotype. Individuals with the SEPP1 25191 G/A genotype (rs3877899) constituted 44% of the cohort and had significantly lower (by 11%) serum SEPP1 levels than those with the SEPP1 25191 G/G genotype, which constituted a similar portion of the cohort. Individuals with the SEP15 811 T/C genotype (rs5845) constituted 31% of the cohort and had significantly lower (by 15%) buccal cell Se levels than those with the most prevalent (65%) SEP15 811 C/C genotype. No other differences were found in the values of Se biomarkers due to these genotypes, or due to GPX4 genotype (rs713041).
The available biomarkers of Se status have been developed for use in individuals suboptimally nourished with respect to the element. Indeed, they have been found generally useful in such cases, as differences in their values tend to correlate with differences in Se intake. More importantly, two of those biomarkers have direct functional significance: GPX3 participates in antioxidant function; while SEPP1 is involved in Se transport and also has antioxidant properties. However, these biomarkers are not informative of Se status under conditions of Se intake exceeding the level necessary to support their maximal expression, which coincides with the level apparently effective in reducing cancer risk.
This study demonstrates the problem of characterizing the Se status of a cohort that is not deficient in the element. It shows the discordance of biomarkers of Se status in a cohort of healthy Americans and the lack of utility of biomarkers with functional significance to assess Se status in a Se-adequate population. The most commonly used biomarker of Se status, plasma Se content, is presumed to reflect both the amounts of Se in various body pools and the level of Se intake [19]. At 142.0 ± 23.5 ng/ml, it showed that this cohort was of relatively high Se status. Half of subjects had plasma Se levels comparable to those of the upper quintile of the NHANES 2003-2004 cohort [11], and 6% had plasma Se levels in the range achieved only by Se-supplementation in the NPC trial [7]. No subject had a plasma Se level <70 ng/ml, above which Nève [12] noted subjects do not show GPX3 responses to Se-supplementation. Accordingly, plasma Se levels, all of which were above that threshold, were not associated with differences in GPX3 activity or SEPP1 level. This indicates maximal expression of those selenoproteins, rendering their measurement uninformative as biomarkers of Se status in this cohort. Differences in SeMet intake affect only the non-specific component of plasma Se, is also likely applicable to other such populations of relatively high Se status
Hill et al [13] estimated that GPX3 and SEPP1, when maximally expressed, should account for about 80 ng/ml of the Se in human plasma. Their estimate was based on the amount of Se associated with SEPP1 in one apparently Se-adequate individual, and the amount of Se reported by Avissar et al [11] to be associated with GPX3. The GPX3 activities and SEPP1 levels of our cohort indicate a slightly lower value, approximately 73 ng/ml, which may reflect weaknesses in our assumptions, particularly those concerning SEPP1. From the reports of Steinbrenner et al [36] and Méplan et al [37], we assumed the presence of multiple SEPP1 variants with an average molecular weight of 60 kD. We also assumed that SEPP1 contained an average of 9.9 g-atoms Se as SeCys per mole based on the findings of Saito et al [35] with human SEPP1; although the genetic coding of the protein would suggest the possibility of as many as 10 SeCys residues per mole [38]. Our estimate suggests that the amount of Se present as SeCys in GPX3 and SEPP1 comprised about 54% of the total amount of Se in the plasma of these non-deficient subjects, a lower percentage than observed (80%) by Deagen et al [39] for Se-deficient men in China. The differences between these different cohorts may reflect the preferential response of the non-specific plasma pool to SeMet. This phenomenon is indicated by nearly half (47%) of plasma Se occurring in the non-specific fraction in the present cohort. Indeed, the variation we observed in plasma Se level was almost exclusively limited to variation in the non-specific component. That this non-specific fraction was not confounded by such variables such as age, gender, metabolic body weight or methylation status (serum folate, vitamin B12, Hcy) suggests that it may be a useful biomarker of Se status in non-deficient populations; since differences in SeMet intake affect only the non-specific component of plasma Se, these findings are also likely applicable to other such populations of relatively high Se status.
We are not aware of buccal cells, which offer the advantage of sampling a metabolically active tissue, having been used previously to assess somatic cell Se. We found buccal cells to contain appreciable amounts of Se; however, their Se showed no significant correlation with plasma Se level. The levels of Se in buccal cells and urine were each positively associated with the use of nutritional supplements. In this regard, these biomarkers may be more useful than plasma Se, which did not show such an effect.
Urinary Se, which consists mostly of methylated selenosugars [40–42], was the only Se biomarker not in dynamic equilibrium with other pools of Se in the body, although it would be expected to show first-order relationships with such pools. That urinary Se was also positively associated with serum folate and vitamin B12, but negatively associated with serum hCys, is consistent with its metabolic production being dependent on the availability of methyl groups. This is supported by the findings of Gonzalez et al. [43] that serum Se level was positively associated with serum folate level and negatively associated with serum hCys level, the latter explaining nearly 6% of the variance of serum Se. The lack of a relationship between plasma Se and urinary Se suggests that these two pools are not in a first-order relationship. Because the non-specific Se was the only variable component of plasma Se, it's apparent that very little of that protein-bound pool turns over into urine. The dimorphic Se excretion between men and women adds to the list of sex-specific differences in (hepatic) Se metabolism, which has been observed in both rodents and humans [44].
The relationship of serum SEPP1 and BMI (r2 = 0.054; p < 0.001), with greatest values among subjects with BMIs of 25-30 (figure 4), was unexpected as Kimmons et al [45] noted low plasma Se levels (<100 ng/ml) somewhat more frequently among women in this BMI class in the NHANES III cohort. That this may reflect the dysregulation of gluconeogenesis in obesity is suggested by studies in cultured cells that have shown SEPP1 to be regulated as a gluconeogenic enzyme [46–48].
It is also clear that genetic variability contributes to variance of Se biomarkers. Of the four allelic selenoprotein variants studied three, GPX1, SEPP1 and SEP15, were significantly related to the values of a Se biomarker (Table 4). That individuals with GPX1 679T/T alleles showed significantly lower plasma Se levels than those of the C/C alleles is of particular interest, as the former genotype has been associated with increased risk to cancers of the lung [49] and breast [50]. That some SEPP1 genotypes have less (11%) SEPP1 expression than others, and that low BMI individuals had significantly lower SEPP1 than those with mid-level and high BMI, it is possible that SEPP1 in some individuals in this cohort may not be maximally expressed, even at these apparently adequate levels of Se intake.
The estimated intakes of macronutrients were similar to those reported for other cohorts of Americans [24]. However, estimated Se intake did not significantly correlate with any Se-biomarker. This is not surprising, considering the inherent errors in determining nutrient intakes from an FFQ and in estimating the amount of Se in particular foods, which can vary considerably depending on the location and/or means of food production [51]. The results of the FFQ method suggested that a core group of 22 foods provided 80% of the Se consumed by this cohort, a number similar to that estimated by Schubert et al [23] for the American population. This core included pork, beef and wheat products, the Se contents of which Finley et al [52, 53] showed can vary enormously (by 3-, 11- and 57-fold, respectively, for items purchased in the upper Midwest). Such great uncertainty severely compromises the value of Se intake estimated in this way as a useful indicator of Se status.
The assessment of Se status in individuals that are not deficient in Se calls for the use of non-classical/unconventional parameters that may be informative despite their limited direct functional significance. In a cohort of healthy Americans, Se intake, estimated using accepted FFQ methodology, was not associated with any biomarkers of Se status; the high degree of uncertainty inherent in this approach rendered it unsuitable for predicting Se status. Two widely used Se biomarkers, GPX3 and SEPP1, showed little correlation with one another. Each appeared to be maximally expressed, neither being associated with total plasma Se. In contrast, Se in the non-specific component of plasma was positively associated with total plasma Se.
It is clear that plasma Se comprises the Se specifically incorporated as SeCys in GPX3 and SEPP1, as well as Se present as SeMet incorporated non-specifically into the plasma proteins. One would also expect plasma also to contain small amounts of Se-metabolites some of which may be noncovalently associated with plasma proteins. In this cohort the non-specific pool was the dominant component, comprising 47% of plasma Se and representing most of the variance in that biomaker. It would therefore appear to be the most useful parameter of Se status in such a cohort.
What we have called the non-specific component of plasma Se was imputed from our measurements of other biomarkers. In order to use it for assessing Se status in non-deficient individuals, it will be necessary to assess it directly. This will mean developing means to assess its various components, which we would expect to include mostly protein-bound SeMet, but also smaller amounts of Se bound to protein-thiols and non-protein bound Se-metabolites including selenosugars.
These results demonstrate that factors other than Se intake that can contribute to variance in Se biomarkers used to assess Se status. Such factors include genotype, methyl-group status, and BMI. It is possible that these factors may contribute to heterogeneity in biomarker responses to Se-supplementation, a problem noted by Ashton et al [54].
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Saito Y, Sato N, Hirashima M, Takebe G, Nagasawa S, Takahashi K: Domain structure of bi-functional selenoprotein P. Biochem J. 2004, 381: 841-846. 10.1042/BJ20040328.
Steinbrenner H, Alili L, Stuhlmann D, Sies H, Brenneisen P: Post-translational processing of selenoprotein P: implications of glycosylation for its utilisation by target cells. Biol Chem. 2007, 388: 1043-1051. 10.1515/BC.2007.136.
Méplan C, Nicol F, Burtle B, et al: Relative abundance of selenoprotein P isoforms in human plasma depends on genotype, Se intake, and cancer status. Antioxid Redox Signal. 2009, 11: 2631-2640. 10.1089/ars.2009.2533.
Hill KE, Lloyd RS, Burk RF: Conserved nucleotide sequences in the open reading frame and 3' untranslated region of selenoprotein P mRNA. Proc Natl Acad Sci USA. 1993, 90: 537-541. 10.1073/pnas.90.2.537.
Deagen JT, Butler JA, Zachara BA, Whanger PD: Determination of the distribution of selenium between glutathione peroxidase, selenoprotein P and albumin in plasma. Anal Biochem. 1993, 208: 176-181. 10.1006/abio.1993.1025.
Gammelgaard B, Madsen KB, Bjerrum J, et al: Separation, purification and identification of the major selenium metabolite from human urine by multi-dimensional HPLC-ICP-MS and APCI-MS. J Anal Atomic Spectrom. 2003, 1: 65-70.
Ogra Y, Ishiwata K, Takayama H, Aimi N, Suzuki KT: Identification of a novel selenium metabolite, Se-methyl-N-acetylselenohexosamine, in rat urine by high-performance liquid chromatography--inductively coupled plasma mass spectrometry and--electrospray ionization tandem mass spectrometry. J Chromatogr B Analyst Technol Biomed Life Sci. 2002, 767: 301-312. 10.1016/S1570-0232(01)00581-5.
Kobayashi Y, Ogra Y, Ishiwata K, Takayama H, Aimi N, Suzuki T: Selenosugars are key and urinary metabolites for selenium excretion with the required to low-toxic range. Proc Natl Acad Sci USA. 2002, 99: 15932-15936. 10.1073/pnas.252610699.
Gonzalez S, Huerta JM, Fernandez S, Patterson AM, Lasheras C: Homocysteine increases the risk of mortality in elderly individuals. Br J Nutr. 2007, 97: 1138-1143. 10.1017/S0007114507691958.
Stoeder M, Renko K, Hög A, Schomburg L: Selenium controls the sex-specific immune response and selenoprotein expression during the acute-phase response in mice. Biochem J. 2010, 429: 43-51. 10.1042/BJ20091868.
Kimmons JE, Blanck HM, Tohill BC, Zhang J, Khan LK: Multivitamin use in relation to self-reported body mass index and weight loss attempts. Med Gen Med. 2006, 8: 3-10.
Burk RF, Hill KE: Some properties of selenoprotein P. Biol Trace Elem Res. 1992, 33: 151-153. 10.1007/BF02784004.
Speckmann B, Walter PL, Alili L, et al: Selenoprotein P expression is controlled through interaction of the coactivator PGC-1alpha with FoxO1a and hepatocyte nuclear factor 4alpha transcription factors. Hepatol. 2008, 48: 1998-2006. 10.1002/hep.22526.
Speckmann B, Sies H, Steinbrenner H: Attenuation of hepatic expression and secretion of selenoprotein P by metformin. Biochem Biophys Res Commun. 2009, 387: 158-163. 10.1016/j.bbrc.2009.06.143.
Ratnasinghe D, Tangrea JA, Andersen MR, et al: Glutathione peroxidase codon 198 polymorphism variant increases lung cancer risk. Cancer Res. 2000, 60: 6381-6383.
Hu YJ, Diamond AM: Role of glutathione peroxidase 1 in breast cancer: loss of heterozygosity and allelic differences in the response to selenium. Cancer Res. 2003, 63: 3347-3351.
Combs GF: Selenium in Global Food Systems. Br J Nutr. 2001, 85: 517-547. 10.1079/BJN2000280.
Finley JW, Mattys L, Shuler T, Korynta E: Selenium content of foods purchased in North Dakota. Nutr Res. 1996, 16: 723-728. 10.1016/0271-5317(96)00062-0.
Keck AS, Finley JW: Database values do not reflect selenium contents of grains, cereals and other foods grown or purchased in the upper Midwest of the United States. Nutr Res. 2006, 26: 17-22. 10.1016/j.nutres.2005.11.009.
Ashton K, Hoopere L, Harvey LJ, Hurst R, Casgrain A, Fairweather-Taite SJ: Methods of assessment of selenium status in humans: a systematic review. Am J Clin Nutr. 2009, 89: 2025S-2039S. 10.3945/ajcn.2009.27230F.
The authors gratefully acknowledge the expert contributions of the following members of the Grand Forks Human Nutrition Research Center staff: Brenda Ling for subject recruiting; Wesley Canfield MD, for subject health oversight; Emily Nielsen RN, Doreen Rolshoven LPN, Judy Schumacher RN, and Ruth Christianson LPN, for coordinating subject visits and sample collection; Sandy Botnen, Teresa Numedahl, Wendy Mayer, Pat Wilson and Kay Williams for sample preparation and clinical biochemical analyses; Bonnie Hoverson, Elvira Bell, Sue Sherette, Charlene Kuntz and Doris Zidon and Mary Jo Peltier for preparation and distribution of supplement packets; Laura Idso for sample preparation and enzyme analyses; and Craig Lacher and Bill Martin for Se analyses.
Additional information
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
The authors' responsibilities were as follows: GFC, JCW, CDD and JAM designed the research; GFC, JDW, MIJ, HZ, AJS, EOU, LS, AH and CSH conducted the research; LKJ analyzed the data; GFC and MIJ wrote the paper and had responsibility for its final content.
The authors read and approved the manuscript.
Gerald F Combs Jr, Jennifer C Watts, Matthew I Jackson contributed equally to this work.
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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Cite this article
Combs, G.F., Watts, J.C., Jackson, M.I. et al. Determinants of selenium status in healthy adults. Nutr J 10, 75 (2011). https://doi.org/10.1186/1475-2891-10-75
DOI: https://doi.org/10.1186/1475-2891-10-75
- Food Frequency Questionnaire
- SeMet
- Buccal Cell
- Metabolic Body Weight
- Serum hCys


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Rising Star: Connecticut Guard Ryan Boatright
By Andrew Kahn
Shabazz Napier and DeAndre Daniels jump off the stat sheets from Connecticut's wins at Madison Square Garden this past weekend. The play of junior guard Ryan Boatright was just as important. On Friday, after Iowa State had cut a 17-point deficit to four with two minutes left, Boatright made what his coach called the play of the game. Driving right off a ball screen, he somehow spotted Niels Giffey in the left corner and contorted his body to snap a crisp pass that led to a back-breaking three.
Boatright hit some big shots of his own against Michigan State in Sunday's regional final, such as his three to push the lead to 10 in the second half, but it was his defense that was critical. Michigan State's senior point guard Keith Appling only scored two points and had twice as many turnovers (four) as assists. When the Spartans turned to Travis Trice for an offensive boost, Boatright smothered him. Trice was 0 for 4 from the field and had to be taken out of the game because Boatright made it difficult for him to get the ball up the court.
"[My coaches] tell me all the time that I'm the defensive stopper for the point guards," said Boatright, who has six steals in the past two games. "I take pride in my defense, picking them up 94 feet, the entire length of the floor. Even if I can't pick a steal, just turning them and turning them and getting them uncomfortable so they can't run the offense so smoothly. So even though my shots weren't falling today, I was going to do something to impact the game and today it was the defense."
>>More College Basketball Rising Stars
Boatright has an inconsistent release on his shot—he's shooting 38 percent this season—and, earlier this season, had a tendency to showcase his exceptional handle at the expense of ball movement, but he's playing his best basketball in the Tournament. He scored 16 on 4 of 10 shooting against Iowa State and scored 11 against the Spartans. Despite his size—he's generously listed at 6-feet, an inch shorter than his backcourt mate Napier—he averages 3.4 rebounds on the year.
"Ryan is growing up," said UConn coach Kevin Ollie, a former point guard who is very passionate about what he expects from that position. "Ryan is allowing us to coach him now. He's opening up and trusting us more. He's meant a lot to us, not only in the games you see [on TV] but what he's done in practice, being more vocal, being a leader." Ollie praised Boatright's unselfishness, and it has been clear during this Tournament run that he's been looking to distribute to the hot hand.
UConn will face No. 1 overall seed Florida in the Final Four on Saturday. The Huskies beat the Gators at home on Dec. 2 on a Napier buzzer-beater. Boatright had nine points and four assists in that game, but it was UConn's work on the other end of the floor that stood out. "I just remember a lot of scrambling on defense," he said after beating Michigan State. "We were everywhere—helping each other, closing out right, and rebounding the basketball. When we got to go out there and play them [again], we've got to play great defense."
Andrew Kahn is a contributor to CBS Local Sports who also writes for Newsday and The Wall Street Journal. He writes about college basketball and other sports at AndrewJKahn.com. Email him at email@example.com and follow him on Twitter at @AndrewKahn
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Happy New Year! I'm afraid my seminar reports are still queued awaiting certain vital feedback before the next one can go up, so instead here's something I've had ready to write for ages. The subject header is, perhaps sadly for our societies, not a phrase one hears often, but happily for you my readers, it is completely appropriate to the subject of this post. That subject is a charter that I read while pulling together a comparative section for my chapter in the volume Allan Scott McKinley and I are editing from the Leeds conference sessions we used to run, now in press.1 The chapter has a substantial section setting pre-Catalan documentary phraseology against that used in its contemporary Asturias-León. This, of course, takes me into the territory inhabited by the expertise of Wendy Davies and Graham Barrett, and in fact I'd heard Graham talk about this charter at Kalamazoo some time ago and then again more locally and recently, as it forms one of a group of documents that tell us that certain counts of the Leonese court took it upon themselves to start bringing public suits against adulterers, adulterers who then often had to pay off the quite unpayable fines by giving lands to the counts. Kalamazoo papers are short, and one has to be selective about what one includes, and that is the only reason I can imagine why Graham would not have told this story himself then—and he may have done in his thesis, even now nearing completion—but, there is more than he told and that more is substantially CHEESE. What do I mean? Well, read this translation.2 It's a bit rough, because the original is not the smoothest, and I've only modernised a few of the names where I'm sure what modern forms would be, but you'll get the idea.
In the name of the Lord. I, Letasia, am infamous to many, indeed it is most well-known to many people that I mixed myself up in adultery with a slave of Hermenegildo, Ataulfo by name, who was holding a tenement of his, and we ate four cows of the animals there and sixty cheeses in secret and they led me before the judge, namely Bishop Froarengo. And the selfsame judge decided that I should pay for those same cows and cheeses twofold, and I was to make over eight acceptable cows and a hundred and twenty cheeses, the which judgement left me well-pleased. On this account it has pleased me, Letasia, for all of this crime which I have professed before the selfsame judge, and thus I pay to you Hermenegildo the whole inheritance I have in the villa where my father Cristobal or my grandparents Abolino, Deodatis and Violicus lived, in the territory of Tamara, that is, land, fruit-trees and all kinds of fruits, meadows, pastures, water-meadows, waters with all buildings or whatever is for the use of men. Thus, so that from this day and time today it be erased from my right and handed over and conceded to your right and you may have power fully in God's name. If, however, any man, what I do not believe shall be brought about, should come against this my act to disrupt it, let him pay you two pounds of gold, and you have it in perpetuity. This little charter of payment or agreement made the 8th Kalends of September, Era 896. Letasia, in this testamentary or judicial scripture, have made the sign of my hand. Sisibert, witness. Savarigo, witness. Assiulfo, witness. Daco, witness. Ebregulfo, witness. Mirello, witness. Ostouredo, witness. Quirico, witness. Ermorico, witness.
I mean, I grant you there are all kinds of interesting implications of language and social practice here. It's more or less built out of formulary phrases without much attempt to get them joined up into sense, but obviously they have been chosen for the job even so. Letasia's husband is not mentioned; one might expect him to be, really, if there were one, which suggests that there wasn't, but the crime is still adultery. Nonetheless, she was not actually required to compensate for the adultery, which was presumably not considered worth punishing; it would have been hard to argue, perhaps, that it had cost Hermenegildo anything except a few hours of his slave's labour (ahem) but for the, well, inconspicuous consumption of four head of cattle and sixty cheeses. I mean, how long was this going on? It's not a one-off, is it, and even a four-off involves enough cheese per person that they would have been pretty easy to catch. Letasia may indeed have been pleased by the judgement, as she could according to the Visigothic Law that still ran here have been put to death or enslaved herself, although not to Hermenegildo but to her own heirs.3 Nonetheless, though she had got away lightly, she had eaten more than she cared to pay back four times over, which gives us some idea how much of a hit Hermenegildo had been able to take without, apparently, noticing. In other words, we're looking here at lifestyles of the rich and infamous in ninth-century Galicia, and those lifestyles on this occasion included a certain amount of sexual impropriety and some seriously big amounts of cheese. We have proof!
1. To my current understanding this can be cited as J. Jarrett, "Comparing the Earliest Documentary Culture in Carolingian Catalonia" in Jarrett & A. S. McKinley (edd.), Problems and Possibilities of Early Medieval Charters, International Medieval Research 19 (Turnhout forthcoming).
2. I'm quoting this from Antonio Cumbreño Floriano (ed.), Diplomática española del periodo Astur. Estudio de las fuentes documentales del Reino de Asturias (718-910), 2 vols (Oviedo 1949), doc. no. 68, but it has been more recently edited in Pilar Loscertales de Valdeavellano (ed.), Tumbos de Monasterio de Sobrado de los Monjes, 2 vols (Madrid 1976), doc. no. 75. The text as Floriano gave it is: "In Dei nomine. Ego Letasia manifesta quidem sum multis, set et multis manet notissimum, eo quod commiscui me in adulterio cum servo Hermenegildi, nomine Ataulfo, qui eius bustum tenebat, et comedimus de ipsis animalis IIIIor vaccas Lxa caseos furtim et adduxerunt me ante iudicem nomine Froarengum episcopum. Et ipse iudex iudicavit ut parierem ipsas vaccas et ipsos caseos in duplum, et facerem octo vaccas placibiles, et centum viginti caseos, quod Iudicum bene mihi complacuit. Ob inde placuit mihi Letasia, ut pro omni ipso furto, quod ante ipsum iudicem manifestavi, pariarem tibi Hermenegildo omnem meam hereditatem integram quam habeo in villa ubi pater meus Christovalus habitavit sive tionis mei Abolinus, Deodatis et Violicus habitaverunt, in territorio tamarense, id est, terras, pumares et omnia genera pomorum, pratis, pascuis, paludibus, aquas cum omnibus edificiis vel quicquid ad prestitum hominis est. Ita ut de hodie die et tempore de meo iure abrasa et tuo iuri sit tradita atque concessa et plenam in Dei nomine habeas potestatem. Si quis tamen homo, quod fieri non credo contra hunc meum factum ad irrumpendum venerit pariat tibi auri libras duas, et tibi perpetim habituram. Facta cartula pariationis vel placiti viiio Kalendas Septembris, era DCCCa LXXXX VIa. Letasia in hac scriptura testamenti vel placiti manu mea signum feci (signum). Sisibertus testis (signum). Savarigus testis (signum). Assiulfus testis (signum). Daco testis (Signum). Ebregulfus testis (signum). Mirellus testis (signum). Ostouredus testis (signum). Quiricus testis (signum). Ermoricus testis (signum)."
3. That said, Letasia's case, as an apparently-freeborn woman with no husband messing with somebody else's slave but clearly at her will and with no intent to marry him, is hard to find an exact ruling for in the Law. The closest fit, whence I get the enslavement idea, seems to be Karl Zeumer (ed.), Leges Visigothorum, Monumenta Germaniae Historica (Leges Nationum Germanicum) I (Hannover 1902, repr. 2005), online here, transl. S. P. Scott as The Visigothic Code, 2nd edn. (Boston 1922), online here, Book III Title IV cap. xiv.


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This sample demonstrates how a model is generated out of a dataset using the Support Vector Machine (SVM) algorithm using tuned hyper parameter values. You can find these parameter values in the
<ML_HOME>/samples/tuned/svm/hyper-parameters file. The sample uses a data set to generate a model, which is divided into two sets for training and testing.
Follow the steps below to set up the prerequisites before you start.
- Download WSO2 Machine Learner, and start the server. For information on setting up and running WSO2 ML, see Getting Started.
- Download and install jq (CLI JSON processor). For instructions, see jq Documentation.
- If you are using Mac OS X, download and install GNU stream editor (sed). For instructions, see GNU sed Documentation.
Executing the sample
Follow the steps below to execute the sample.
Navigate to
<ML_HOME>/samples/tuned/svm/directory using the CLI.
- Execute the following command to execute the sample: ./
Output of the sample
Once the sample is successfully executed, you can view the summary and the prediction of the model as described below.
By default , the sample generates the model in the
<ML_HOME>/models/ directory of your machine. For example, the generated file is in the following format denoting the date and time when it was generated:
Viewing the model summary
You can view the summary of the built model using the ML UI as follows.
Log in to the ML UI from your Web browser using
admin/admincredentials and the following URL: https://<ML_HOST>:<ML_PORT>/ml
Click the Projects button as shown below.
- Click MODELS button of the new analysis which you created by executing the sample as shown below.
- Click VIEW of the built new model as shown below.
You view the summary of the built model as shown below.
Viewing the model prediction
The sample executes the generated model on the
<ML_HOME>/samples/tuned/svm/prediction-test data set, and it prints the value
] as the prediction result In the CLI logs.


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For months I've been reading about other people's Life Lists and wondering if I could ever make one without driving myself crazy with all the achieving and in-my-head competition.
Would my list be cool enough?
Would I really do everything I said I would?
What huge adventures await?
Inspired by Rosie's birthday list, Sarah's try new things before her birthday list and Amy's Life List, I've decided to put together a list of things I would like to do before I turn 43.
Why 43 instead of 42?
My birthday is in 4 months and I don't want to hurt myself getting it all done before then. My list is l-o-n-g, makes me think I needed to start this thing in my 20s.
My intention with this list is to remind myself to have more fun, take care of myself and to do things out of the ordinary.
I'll keep you posted on my progress.
The List:
1. Take my birthday (March 20th) off – no work, no obligations – just fun.
2. Take an anti-gravity yoga class.
3. Road trip to Portland with Ruby.
4. Take Ruby to see live music.
5. Read 43 books.
6. Put my feet in the ocean with Ruby.
7. Pick berries.
8. Go on retreat.
9. Teach an active yoga class.
10. Go to the movies by myself. {Yep, I've never done this. I have, however, gone to a rock show on my own so don't feel too sorry for me.}
11. Attend a service at a Universal Unitarian church.
12. Explore the Taiwanese culture in Sacramento.
13. Learn 10 words/phrases in Mandarin.
14. Run a 5K.
15. Write a fan letter.
16. Rent a house at the coast or in the mountains.
17. Teach a class on social media and blogging to teachers.
18. Sleep outside.
19. Do a swim workout.
20. Watch my first students graduate from high school.
21. Spend a week away from the Internet.
22. See a play.
23. Go to a sing along at the Crest.
24. Take a photography class.
25. Take a cooking class.
26. Take a private yoga class.
27. Actually make something I've pinned on Pinterest.
28. Go to the ballet.
29. Have a family portrait taken.
30. Watch a movie outside in the backyard.
31. Take Ruby on a hike.
32. Help a friend start a blog.
33. Build an outdoor shower.
34. Make a wall of bookshelves in the dining room.
35. Redesign the entryway and make it pretty and useful.
36. Start a mom/play group in Midtown.
37. Teach a class at my gym.
38. Organize the pantry.
39. Get rid of all the shoes I don't wear because they are uncomfortable.
40. Take a nap in a hammock.
41. Host a baby shower.
42. Take a photo every day.
43. Make the Book of Me into a book.
Do you make yearly plans? A Life List? Would you like to?
I'd love to hear about yours.
And I'd love for you to join me in making my list a reality. Please let me know if you'd like to do any of my 43 with me.


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Things to Have In Mind When Hiring Home Rebuilding Service
When moving into a new house, remodeling services are important to improve its state to meet your standards. Only employing remodeling experts on your job can see you through your project's objectives. There are qualified renovating tradesmen within you that can offer you quality advice on the right fixtures to employing on your job. You need to be clear on the qualities your renovation contractor must possess because many of them post the same working qualities on their company profile. Therefore, it is important to consider professional help from people that sourced this services awhile before you because they have experience on the whole operation. You need to evaluate company profiles owned by the services present to take up your job to determine whether their working qualities meet all your standards. There are important things you need to have in mind when choosing a renovator for your home. Therefore, here are the key factors for you to consider when choosing a home renovator.
The cost of operation of your remodeling service deserves great deliberations. All the remodeling contractors will not offer you the same cost quotes on the same job you have on board. In this case, it is important to be certain on the payment options adopted by the home remodeling service of your choice because these are the exact rates they will employ on your job. You need to stray from low-cost quotes because many of them are posted by home renovators whose motives are destined to lure potential clients on their extortion plan while deep down their working skills are wanting. You need to come up with a budget that will help you have a clear mind on the right resources to allocate for your job.
You need to ponder the distance between you and the home rebuilding service of your choice. A home rebuilding contractor that great understanding on the local industry will be effective because they will not strain to try to locate reputable suppliers for your fixtures. All the home remodeling contractors want to market their quality output to other local potential clients and only good work can acquire great sells in the market and considering them will offer you a guarantee of premium-quality services. There are people within you that you can rely on for quality information regarding the quality of service output offered by the renovator of your choice in order to know whether you are hiring the right service.
You need to ponder the expertise of your home remodeling service. Excellent verbal and written communication skills that helps them interact effectively with the client and other co-workers to analyze the whole rebuilding situation to come up with a perfect solution.
How I Achieved Maximum Success with
Looking On The Bright Side of