All values are log 2 fold change over untreated, time-matched controls. To determine whether this effect was unique to HCT cells, we also treated three other cell lines derived from diverse human tissues. Thus, the ability of propionate and butyrate to increase histone acetylation, but not methylation, can be generalized to multiple cell types. This experiment raised several intriguing questions. First, what differentiates propionate and butyrate from acetate?
It is general knowledge that butyrate can act as an HDAC inhibitor, slowing the removal of acetate from chromatin Davie, This could explain the difference between acetate and butyrate, but it does not necessarily explain the difference between acetate and propionate. While SCFA concentrations in media were theoretically higher than our determined IC 50 values Figure 2C , it has been previously reported that intracellular concentrations of SCFAs are much lower than extracellular concentrations, which could have bearing on this mechanism Donohoe et al.
The calculated values suggest that free SCFAs are rapidly metabolized instead of building up in cells Boets et al. With this in mind, we searched for labeled carbons in downstream TCA cycle metabolites. However, we found no significant labeling of TCA cycle metabolites after propionate and butyrate treatment Figure 2D. Instead, labeled carbons built up as acyl-carnitine species, appearing in as little as 10 min and remaining stable for 24 hrs Figure 2E.
Intriguingly, this buildup did not occur in acetate-treated cells. The same trend occurred in acyl-CoA levels, although these species degrade rapidly during sample processing and are thus difficult to detect by mass spectrometry Figure 2—figure supplement 1. In that study, increases in acyl-CoA lead to a decrease of free coenzyme A. Under our conditions, however, free carnitine and free CoA levels did not significantly change Figure 2—figure supplement 1. All cell lines showed the same upregulation of acyl-carnitines after butyrate and propionate treatment Figure 2—figure supplement 1.
Ultimately, these data suggest that histone hyperacetylation may not be due to the presence of intracellular propionate or butyrate, as the intracellular levels of these metabolites are low, but to the presence of propionyl- or butyryl-CoA. Since propionyl- and butyryl-CoA do not inhibit HDAC at physiological concentrations, this raises the possibility that the histone hyperacetylation phenotype may not be due to HDAC inhibition, but to a different mechanism Table 1 ; Vogelauer et al.
To test whether this occurred in our system, we performed histone proteomics after a 4-hr treatment with 13 C-labeled SCFAs. This data indicates that acyl-CoAs are not metabolized to acetyl-CoA in sufficient quantities to explain the induced hyperacetylation, indicating a different mechanism is at play. Next, we tested whether the rapidly formed propionyl- and butyryl-CoA from propionate and butyrate treatment could directly activate HATs Table 1.
Free propionate and butyrate had no effect. However, since this assay simply measures the release of CoA, the apparent increase in HAT activity could have resulted from increased propionyl- or butyryl-transfer reactions. In addition, these results do not indicate which HAT enzymes may be activated. C Rate of histone acetylation by acetylated, propionylated, or butyrylated p using radioactive acetyl-CoA.
Replicate results from this assay performed with different enzyme preparations on different days are shown in Figure 3—figure supplement 1. Quantification for all replicates including Western blots and radioactive assays available in Source data 1. Most HATs prefer specific histone lysines and catalyze acetyl-transfer to a handful of histone substrates Roth et al.
Both proteins encode an autoinhibitory loop AIL, also called the autoregulatory or activation loop , which must be auto-acetylated to stimulate HAT activity Karanam et al. Large-scale mass-spectrometry surveys have identified sites of propionylation and butyrylation on p, including on several AIL lysines; however, how these modifications are added and whether they have a functional consequence on p activity are not known Chen et al. First, we determined whether unmodified p can catalyze auto-acylation using propionyl- or butyryl-CoA.
Indeed, we found that recombinant p performed auto-acylation with propionyl- or butyryl-CoA at rates indistinguishable from auto-acetylation with acetyl-CoA Figure 3B. Auto-acylation was catalytic, as heat-denatured p did not show increases in acylation under the same conditions Figure 3—figure supplement 1A.
Proteomics analysis confirmed the location of propionylation and butyrylation on several AIL lysine sites under these conditions Table 2. Sites of lysine acetylation, propionylation, and butyrylation on recombinant p treated with acyl-CoAs. Next, we investigated if auto-acylation activates p toward histone acetylation. All forms of acylated p acetylated, propionylated, and butyrylated were significantly more active than an unmodified control Figure 3C.
Again, these data showed an increase in activity for all three forms of acylated p In all, these data demonstrate that p catalyzes auto-butyrylation and propionylation on AIL sites, and that auto-acylation activates the enzyme in a similar manner to auto-acetylation. To investigate whether this previously unappreciated mechanism functions in vivo, we used pharmacological inhibitors of each potential pathway. First, we treated cells with a potent inhibitor of p, A A inhibits both histone acetylation and auto-acetylation of p, and thus should inhibit histone hyperacetylation due to p activation Lasko et al.
A treatment did not just reverse SCFA-induced hyperacetylation on histone substrates of p, but also on the non-histone p substrate p53 K Thus, if propionate and butyrate act solely as HDAC inhibitors, they should not induce further hyperacetylation under these conditions. Collectively, these results suggest that propionate and butyrate induce histone hyperacetylation by an alternative mechanism to HDAC inhibition, likely through the auto-activation of p by rapidly formed acyl-CoAs.
Values are normalized to total H3 or total p53 before calculating fold changes to the appropriate untreated control. Values are fold change over untreated cells with the same transfection. Activity is measured with a radioactive assay and normalized to concentration of immunoprecipitated p in each sample. Nevertheless, these data strongly suggest that the AIL lysines of p are necessary to induce the full extent of histone hyperacetylation after propionate treatment.
Corresponding western blots also identified increased propionylation and butyrylation of p under these conditions Figure 4—figure supplement 1D. In all, our results demonstrate that p activation, rather than HDAC inhibition, plays a critical role for propionate and butyrate-induced chromatin hyperacetylation at low concentrations.
While it is possible to reach those concentrations in the gut lumen or portal vein, few other organs would normally reach these values. In addition, our previous data suggests that SCFAs can induce histone hyperacetylation in colon, liver, and adipose tissue, which experience SCFA concentrations that differ by orders of magnitude Krautkramer et al. Even in the gut, intracellular concentrations of SCFAs could be significantly lower than exterior concentrations Donohoe et al.
Thus, the idea that increased histone acetylation is primarily mediated by HDAC inhibition requires re-evaluation. In this study, we report a previously unappreciated mechanism and provide compelling evidence that butyrate and propionate activate p at low levels, through the rapid conversion to propionyl- and butyryl-CoA, catalytic auto-acylation, and subsequent activation of p Hyperacetylation induced by high doses of butyrate 2—5 mM are almost immediately reversed after butyrate withdrawal Prasad and Sinha, ; Wang et al.
In contrast, auto-acetylation of p increases its half-life from 4. Thus, even after extracellular SCFAs have been fully metabolized, modified p could still theoretically induce hyperacetylation. Auto-acylation of p may thus prove a primary mechanism in physiological systems where SCFA concentrations are low.
It is important to note that acetate had negligible effect on histone acetylation in the cell-based systems described in this study. Acetate treatment has been previously reported to increase histone acetylation, but only under extreme conditions such as hypoxia Bulusu et al.
Others have reported no effect of acetate in conditions similar to those described here Hinnebusch et al. Heat shock protein production is a common cellular stress response mounted to mitigate the toxic effects of misfolded proteins in cells and is known to be activated by mistranslating tRNAs in yeast 18 , Increased heat shock protein levels were also observed in mice expressing editing-defective aminoacyl-tRNA synthetases 25 , and mammalian cells expressing mistranslating tRNAs after extended transfection periods of up to 72 h We also measured HSP70 levels after more extended transfections periods 24, 48 and 72 h and observed no significant differences between wild-type and mistranslating cells Supplementary Figure S7B.
Cells can also down-regulate mRNA translation at the level of elongation. Eukaryotic elongation factor 2 eEF2 is involved in ribosome repositioning and movement of tRNA from the A-site to P-site during translation Phosphorylation of the eukaryotic elongation factor 2 eEF2 by eEF2 kinase eEF2K at Thr56 reduces translation elongation rates in conditions of nutrient deprivation or various other forms of cellular stress, including proteasome inhibition with MG MG treatment stimulated a significant increase in p-eEF2 levels in both cell lines Supplementary Figure S8 , but we did not observe any difference in p-eEF2 levels in the mistranslating cells compared to wild type.
In normal cells, MG inhibits both protein degradation see Figure 7C and protein synthesis, leading to no change in the steady state protein levels.
Despite the clear induction of p-eEF2 in MGtreated cells, mistranslating cells show a severely reduced level of protein synthesis in conditions that have high or low p-eEF2 Supplementary Figure S8C. Fluorescent images were captured throughout the time course, and a cytotoxicity assay was completed at the final timepoint. Unlike normal cells, mistranslating cells were unable to increase protein synthesis levels in response to ISRIB. B Fluorescence intensity per cell area of six technical and three biological replicates was quantitated in ImageJ see supplemental methods before and after treatment.
C Cytotoxicity after treatment was measured with CytotoxGlo. Given the synthetic toxic effect we observed in mistranslating cells treated with MG, and previous reports of tRNA anticodon variants promoting increased proteasome activity 40 , we assayed protein turnover in mistranslating cells.
Treatment with MG to inhibit the proteasome was used as a negative control as described In normal cells, MG lead to a reduced rate or protein degradation Figure 7C , while in mistranslating cells MG was not able to further slow their already defective rate of protein degradation Figure 7D.
Protein turnover and clearance of PolyQ aggregates in mistranslating cells. Fluorescence was quantitated in ImageJ see supplemental methods.
Average rate of protein decay B is shown as a change in fluorescence normalized to initial cell area C , D see supplemental information. HTT-exon1 expression was induced with doxycycline 48 h post-transfection and cells were imaged daily by fluorescence microscopy.
After 96 h, Dox was removed and daily imaging was resumed for 72 h. Aggregate counting is described in methods see Supplementary Figure S To assess whether 74Q aggregate clearance was reduced in an independent polyQ model, we used a rat PCderived cell line with a stable, genome-integrated HTT-exon1 74Q fused to EGFP under control of a doxycycline inducible promoter Using mCherry to identify tRNA-transfected cells, we counted the number of transfected cells expressing either wild-type or mutant tRNAs containing at least one aggregate Supplementary Figure S The number of cells containing aggregates was not significantly different during doxycycline treatment, but after doxycycline was removed, aggregates persisted significantly longer in cells expressing the mistranslating tRNA Figure 7E.
Indeed, cells with wild-type tRNA had a significantly reduced number of polyQ aggregates at 48 and 72 h after removal of doxycycline, yet we did not observe any decrease in polyQ aggregates in the mistranslating cells following removal of doxycycline. The data suggest that mistranslating cells are defective in clearing protein aggregates that cause disease. Our data show that huntingtin aggregates form readily in mistranslating cells, but at a slower rate than in wild-type cells.
Since cells expressing the 74Q and mistranslating tRNA strongly suppress protein synthesis, polyQ protein concentrations are less likely to reach the threshold required to seed aggregates We also found that mistranslating cells were generally defective in their ability to degrade proteins.
While mistranslating cells synthesize aggregating proteins more slowly, they are also defective in their ability to degrade huntingtin protein aggregates.
In these cellular models of neurodegenerative disease, the data indicate that natural mistranslating tRNA variants have the potential to affect the onset, progression, and severity of HD. Although spectral counting may overestimate the error rate, even modest levels of mis-incorporation can lead to the accumulation of mis-made protein as we found. Although mistranslating cells were slower in forming HTT-exon1 74Q aggregates, we observed the normally non-aggregating 23Q protein form a significant amount of protein aggregate in mistranslating cells.
In mistranslating cells, we observed 23Q aggregates at 48 h post-transfection. The data suggest that compared to 74Q, higher expression levels and longer times were needed for the 23Q-EGFP protein to aggregate in mistranslating cells. We also observed increased 23Q aggregation in cells treated with the proteasome inhibition. Our observations on huntingtin protein aggregation suggest mistranslating cells are compromised in their ability to both synthesize and degrade protein aggregates.
Our work is the first to explore the interaction between mistranslation and huntingtin polyQ aggregation. Our studies, focused on mistranslation resulting from a tRNA mutant, suggest that other routes to amino acid mis-incorporation, e.
Interactions between other factors regulating protein homeostasis and polyQ aggregation represent a continuing theme in yeast and mammalian cell models of polyQ aggregation and toxicity. For example, work in yeast shows that molecular chaperones 67 , 68 and protein degradation 69 are critical systems to combat polyQ aggregation.
In Drosophila S2 cells expressing long Q, but not shorter polyQ alleles, protein synthesis is downregulated via the translation regulator Orb2 Aggregates were monitored over a 48 h period, and in agreement with our study, no toxicity from the 64Q or even the Q HTT allele was observed In HeLa cells, the same authors used a reporter for nuclear retention of ribosomal protein S2, which signifies inhibition of ribosome biogenesis.
Cells expressing a 97Q allele showed a 3-fold increase in nuclear localization of ribosomal protein S2, demonstrating defective ribosome biogenesis in cells expressing an aggregating HTT allele The authors did not directly measure the rate of protein synthesis, but their data suggest dysregulation of protein synthesis in cells expressing long polyQ alleles. Our data indicate that tRNA variants, which compromise protein homeostasis, may exacerbate the dysregulation of protein synthesis caused by deleterious and longer polyQ alleles.
Using mass spectrometry, we confirmed amino acid mis-incorporation of Ser at multiple Phe codons. Mistranslating cells were characterized by increased cytotoxicity, general inhibition of protein synthesis, sensitivity to proteosome inhibition, and resistance to the neuroprotective stress response inhibitor ISRIB.
The data suggest that some mistranslating tRNA variants can exacerbate polyQ toxicity in a model of neurodegenerative disease. Conversely, the naturally occurring tRNA Ser AAA variant was toxic alone, but did not show additional toxicity with aggregating huntingtin protein. The differences in toxicity may depend on how cells respond to different types of mistranslation.
Furthermore, cells mis-incorporating Ser at Phe codons showed a stronger repression of translation than cells mistranslating Pro codons with Ser. The severely inhibited protein synthesis observed in cells with tRNA Ser AAA may indeed protect the cell from the toxicity of the aggerating huntingtin allele and amino acid mis-incorporation.
A recent study in yeast demonstrated that tRNA variants that result in different types of amino acid change can elicit distinct cellular responses For example, down-regulation of genes involved in cell cycle, DNA replication, transcription, and response to stress was observed in cells mistranslating Pro codons with Ala but not in cells mis-incorporating Ser at Arg codons Phenotypic differences observed from expressing different tRNA variants may also depend on the compatibility of the tRNA in question with a given gain-of-function mutation.
For example, nucleotides in and adjacent to the anticodon sequence are often conserved within tRNA isoacceptor groups 46 , and can be modified to ensure optimal fidelity and efficiency in recognizing certain codon sets.
This is true for wild-type tRNA Phe , wherein modified guanine bases in position 34 73 of the anticodon and the anticodon-adjacent position 37 74 are used to efficiently and accurately decode UUU and UUC codons. Although eIF2a is phosphorylated in our mistranslating cells, their lack of response to ISRIB suggests mistranslating cells are defective in translation regulation.
The synthetic phenotype suggests that mistranslating cells rely critically on protein turnover and activity of the proteasome. We also found that protein turnover rates were reduced in cells expressing tRNA Ser AAA, consistent with an increased burden on the proteasome caused by amino acid misincorporation. In conditions of nutrient deprivation and various forms of cellular stress 62 , cells can also down-regulate elongation through inhibitory phosphorylation of eEF2 76 , which catalyzes ribosomal translocation and repositioning of tRNA from the A-site to P-site of the ribosome While mistranslating cells responded to proteasome inhibition MG by phosphorylated eEF2 similarly to normal cells, expression of tRNA Ser AAA showed a dominant effect on repressing protein synthesis, regardless of stress conditions tested or eEF2 phosphorylation status.
Stressors, including MG, may also alter the activity of the mutant tRNA or the tRNome more broadly, as transcription 78 and modification 79 of tRNAs are regulated in response to stress.
Investigating the interaction between mistranslating tRNAs and the impact of tRNA processing on protein quality control will form the basis of future investigations. Humans encode over tRNA genes.
Human cells and tissues are estimated to express between and different tRNA genes While some of these genes may be redundant in function, others are critical for maintaining protein homeostasis. Despite a vast background of tRNA genes, even a single tRNA mutant has the potential to cause amino acid mis-incorporation, thus exacerbating protein misfolding diseases at the molecular level.
Notably, mistranslating cells exhibit severely inhibited protein synthesis, leading to reduced but effective formation of protein aggregates in cellular models of HD. The mistranslating cells were also defective in clearing huntingtin aggregates and were resistant to the neuroprotective compound ISRIB. Because this compound shows promise as a treatment for neurodegeneration 80 , our studies suggest that an active mistranslating tRNA in a patient's genetic background may contribute to drug resistance.
Taken together, our data show that naturally occurring tRNA mistranslators have significant potential to disrupt protein homeostasis and act as modifiers of neurodegenerative disease. Custom imageJ macros are available in the Supplementary Information.
For western blots, full size images are included as a supplementary data file. We would also like to thank Matthew D. Berg, Christopher J. Brandl and Ilka Heinemann for critical discussions and suggestions on the manuscript.
Barratt at University of Cambridge, UK. Funding for open access charge: CIHR. Crick F. The origin of the genetic code. Google Scholar. Steiner R. Regulation of tRNA-dependent translational quality control. Kunkel T. Rna-sequencing from single nuclei.
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Liu, L. Chen, T. Ko, A. Fields, and E. Oncogene, 25 25 —75, Feb Strategies for microarray analysis of limiting amounts of RNA. Brief Funct Genomic Proteomic, 2 1 —6, Apr The last fifty years then become less about me and more about we.
Less about selfishness and more about service. We stop adding more things into our lives and begin to subtract—and simplify. We learn to savor simple beauty, experience gratitude for small miracles, appreciate the priceless value of peace of mind, spend more time cultivating human connections and come to understand that the one who gives the most is victorious. And this becomes, potentially, your gateway into immortality.
Dad was a remarkable man—a tremendously successful business pioneer. He flew airplanes, raced fast cars and loved superb wine. He was so alive. Anyway, the stress and shock of his world collapsing pushed him to do what we could never have imagined. He placed a hand with a hippie ring on a pinky finger onto his heart as he spoke these words, looking both chivalrous and bohemian.
The homeless man interrupted the intimate moment the two were sharing. Thank you. Dismiss these as idealistic words of an elderly inspirationalist if you wish. Our world needs more of us. And yet, I am also a realist. They secure their identity by who they are externally.
They compare themselves to the orchestrated—and fake—highlight reels presented by the people they follow. They measure their self-worth by their net worth. This explains why the majority is sinking in the quicksand of uncertainty, boredom, distraction and complexity. All they do is complain about how bad things are for them instead of applying their primal power to make things better. They take instead of give, criticize instead of create and worry instead of work. Build antibodies to combat any form of average from getting anywhere near your professional days at the office and your private life at home.
Never be a drama mama. If you were standing there with them, you would think he was weird too. Because leadership is a lot less about having a formal title, a large office and money in the bank.
And a lot more about committing to mastery over all you do—and in who you are. Real leadership is about sending out brave work that exemplifies genius, turns your whole field on its head by its scope, innovation and execution, and is so staggeringly sublime that it stands the test of time.
And never work only for the income. Labor for the impact. Make your dominant pursuit the heartfelt release of value that represents an uncommon magic that borders on the poetic. Develop the patience to stick with your dedication to absolute world-class output, even if over a lifetime you only generate a single masterpiece.
Be a virtuoso. A standout. An exceptionalist. So never mail it in. Always bring it on. The homeless man now had his eyes closed. And was down on the floor doing a series of one-armed push-ups.
I read that Khalil Gibran carried the manuscript around with him for four long years and refined it constantly before giving it to his publisher, just so it was pure art. His words keep me reaching for a greater power as an artist, even though I battle procrastination a lot. But I know I can be great. If I could just beat my self-sabotage. And my demons. Beads of perspiration meandered down his angular face. And if you do so, a guaranteed victory is in your future.
And this brings me to the single most important principle of my talk: The greatest starting point for winning in your work and making a splendid life is joining what I call The 5 AM Club. The entrepreneur was now taking notes with a ferocious intensity not previously seen. The homeless man burped, then got down to the floor and held a plank, the kind fitness pros at the gym love to do to build a strong core.
You could hear The Spellbinder begin to cough even more fiercely. A brutal—and sustained—pause followed. Next, he uttered these words, haltingly. He was wheezing audibly. His voice began to quiver like a novice telemarketer on her very first sales call. Joining The 5 AM Club is the one behavior that raises every other human behavior. This regimen is the ultimate needle mover to turn you into an undefeatable model of possibility.
The way you begin your day really does determine the extent of focus, energy, excitement and excellence you bring to it. Each early morning is a page in the story that becomes your legacy. Each new dawn is a fresh chance to unleash your brilliance, unprison your potency and play in the big leagues of iconic results. You have such power within you and it reveals itself most with the first rays of daybreak.
Please do not allow past pains and present frustrations to diminish your glory, stifle your invincibility and choke the unlimited possibilitarian that lurks within the supreme part of you. In a world that seeks to keep you down, build yourself up.
In an epoch that wishes you would stay in the dark, step into your light. At a time that mesmerizes you to forget your gifts, reclaim your genius. Our world requires this of each of us. To be champions of our crafts, warriors for our growth and guardians of unconditional love—for all of humankind.
Display respect and compassion for all other people who occupy this tiny planet, regardless of their creed, color or caste. Lift them up in a civilization where many get energy tearing others down. Help others sense the marvels that sleep within them. Show us the virtues we all wish more would practice. And, once done, to spend the rest of your days reconnected with it.
Accept this opportunity to human mastery and I promise you that a synchronicity of success as well as an orchestrated magic well beyond the boundaries of logic will infuse the remainder of your days.
And the larger angels of your grandest potential will begin to visit you regularly. Actually, an orderly series of seemingly impossible miracles will descend onto your most genuine of dreams, causing the best of them to come true.
And you will evolve into one of those rare and great spirits who upgrade the whole world by the simple act of walking amongst us. The conference hall was now dark. The entrepreneur let out a sigh the size of Mexico City.
The artist was motionless. The homeless man began to cry. He then stood on a chair, raised his arms like a preacher and boomed these words of Irish playwright George Bernard Shaw: This is the true joy in life, the being used for a purpose recognized by yourself as a mighty one; the being a force of nature instead of a feverish little clod of ailments and grievances complaining that the world will not devote itself to making you happy.
I am of the opinion that my life belongs to the whole community, and as long as I live it is my privilege to do for it whatever I can. I want to be thoroughly used up when I die, for the harder I work the more I live. I rejoice in life for its own sake. It is a sort of splendid torch which I have got hold of for the moment, and I want to make it burn as brightly as possible before handing it on to future generations.
The homeless man then fell to his knees. Kissed his holy beads. And continued to weep. The skill to mold the material into what we want must be learned and attentively cultivated. Let me do this for you cats. Your lives will start to look glorious—within a fairly short time.
And the ride with me will be fun. Not always easy, as we heard from the old guy on the stage. But valuable and prolific and beautiful. Maybe even as wonderful as the ceiling of the Sistine Chapel. He then raised his shirt to show Greek god abdominal muscles. A long finger of a grimy hand moved along the contours, the way a raindrop zigzags down the stem of a rose after a May shower.
Work is how I got all lean and chiseled up like this. Plenty of push-ups, pull-ups, planks, sit-ups and seriously sweaty cardio sessions, often on my special beach. Values few believe in these days, where so many have an entitlement mentality, expecting a rich, productive and fulfilling life to just show up one day like a sparrow at the beginning of spring. And expecting everyone around them to invest the effort they are responsible for inputting. Not judging, just saying. Not complaining, just reporting.
Less talk and more do is what I say. Oh, and check this out. Victors adore education. Some of your most valuable and sensational moments ever will unfold there. You need to take a trust walk with me, people.
And, to be clear, empires arrive in many forms—economic is just one of them. You can also create empires of artistry, productivity, humanity, philanthropy, personal freedom and even spirituality.
You guys can swim in the sea, go snorkeling with the dolphins and fly over the sugar cane stalks that dance in the wind in the helicopter I own. And should you both accept my heartfelt invitation to visit me, I insist you stay at my home. It was becoming increasingly evident that, like many in his field, he was acutely emotional, vigilant to the infinitesimal and carried a sensitivity born of latent pain.
Those who feel more than most people sometimes believe they have been cursed. In fact, they have been granted a gift: one that allows them to sense what others miss, experience the delights that most neglect and notice the majesty in ordinary moments. Yes, such people get hurt more easily, yet they are also the ones who create great symphonies, architect dazzling buildings and find cures for the sick.
And believe me, they named it accurately. Just say yes to life. Like the guru on the platform said, a magic will show up for you the more you start exploiting the terrific opportunities that appear along your path, seemingly by accident. But you need to do your part and go all in when windows of opportunity appear. Oh— and if you come to my home on the island, the only thing I ask is that you stay long enough for me to teach you the philosophy and methodology that my secret adviser shared with me.
Take all the time you need. Maybe even something magical. I know how insane that sounds. And I am fascinated by this mentor he keeps talking about, this teacher who sort of sounds like a modern-day master.
And he obviously seems to have a lot of experience. But just look at him! Man, the guy looks completely down and out—a complete mess. His clothes are all ripped. And sometimes he talks total crazy talk. We have no idea who he is. This could be dangerous. He could be dangerous. Definitely super-weird.
Her eyes still seemed melancholic, though. A daughter growing up without a father is incredibly scary. To be honest, I still carry a lot of the emotional trauma with me. Yes, this made me extraordinarily tough on the outside. And ruthless in some ways. The chip on my shoulder over the loss of my father gave me my drive and my ambition. Yet the loss also left me empty within. I felt inspired when The Spellbinder spoke about never doing something for the money but, instead, reaching for world-class as a leader and a person for the meaning it provides, for the opportunity to grow it provokes and for a shot at changing the world.
His words made me feel so hopeful. I only came to this meeting because my mom gave me a free ticket. I guess I just feel safe with you. His body language showed he was engaged. He no longer anxiously played with his goatee and dreadlocks. And my gut tells me this down-and-out man who wants to teach us how an excellent morning routine can build creative, productive, financial and happiness empires really can help me. And help us. I love that he expresses himself so poetically sometimes and so passionately at others.
He thinks so vividly and quotes George Bernard Shaw like his life depended on it. Really cool. And you and I just met as well. You seem like such a nice person. A few rough edges maybe. I think I understand where those come from. I know it. He glanced at the homeless man, who was eating slices of avocado from a plastic bag. While he was munching on his snack he was also talking on a relic of a mobile phone and staring at the ceiling. A special opportunity to access a whole new universe of originality.
This might be the best thing yet for my art. What truly horrible lives they must lead. Some instinct is also telling me to do this. They both stood up and made their way to the homeless man, who was now sitting with his eyes closed.
Your Mindset is an enormously potent tool for private greatness, prodigious productivity and creative victory—along with your Heartset, Healthset and Soulset. Anyhoo, back to why I closed my eyes. Nearly every morning, I envision my ideal performance for the day ahead. Then I go out and do my finest to live out that perfect day.
Good science is confirming that this practice helps me upregulate my genome by turning on genes that were previously asleep. Not to worry, cats. Oh, in case you were wondering, an SOP is a standard operating procedure. I assume you two are coming? I desperately want to improve my performance and my daily productivity. Take us to your village. Give us some coconuts. Let us ride your dolphins. And improve our lives.
And it will be supported by strong data, the latest research and immensely practical tactics that have been battle-tested in the tough trenches of industry. Get ready for the greatest adventure you cats will ever experience! He looked somewhat surprised by the extent of his graciousness.
All expenses are on me. I thank you. The homeless man smiled tenderly and scratched his beard thoughtfully. You only need a heart full of grace.
A soul generated by love. Increase the state of your fellow human beings and, naturally, your own state of being ascends. Success is cool. But significance is rad.
Generosity—not scarcity—is the trait of all of the great men and women who have upgraded our world. And we need leaders, pure leaders and not narcissists obsessed with their own self-interests, as never before. The two listeners grinned. He then grew quiet.
And let out a deep breath. And most important, have the courage to follow your heart and intuition. They somehow already know what you truly want to become. Set for a new beginning. Such eerie—and sometimes violent—dreams.
This was a terrible idea. It was AM. He was dressed in black with a ruby red polka-dotted bandana on his left wrist.
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