Vitamin B2 (riboflavin) Positively Modulates The Microbiome

[haidut]

I posted a few studies recently showing that riboflavin has an anti-endotoxin effect and protects from colonic inflammation. Apparently, in addition to its direct effects on the TLR4 receptor, riboflavin is capable of actually changing the microbiome composition when used as a supplement. It seems to increase the type of bacteria in the colon that can use riboflavin as an electron carrier, and that type of bacteria is usually not pathogenic. For example, a recent study found that a synthetic analog of riboflavin-5-phosphate (R5P) can eradicate C. Difficile infection that is resistant to antibiotic treatment. R5P is also known as FMN and the study suggests that FMN/R5P would work just as well as the synthetic compound 5FDQD. Unlike treatment with common antibiotics, the eradication of C. Difficile with the riboflavin analog did not affect the counts of other less pathogenic bacteria commonly found in the colon. The HED was just 1mg/kg daily and the treatment lasted for 5 days, but even a single dose worked very well.
Novel riboswitch-binding flavin analog that protects mice against Clostridium difficile infection without inhibiting cecal flora. - PubMed - NCBI
"...Novel mechanisms of action and new chemical scaffolds are needed to rejuvenate antibacterial drug discovery, and riboswitch regulators of bacterial gene expression are a promising class of targets for the discovery of new leads. Herein, we report the characterization of 5-(3-(4-fluorophenyl)butyl)-7,8-dimethylpyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione (5FDQD)-an analog of riboflavin that was designed to bind riboswitches that naturally recognize the essential coenzyme flavin mononucleotide (FMN, also known as R-5-P) and regulate FMN and riboflavin homeostasis. In vitro, 5FDQD and FMN (R-5-P) bind to and trigger the function of an FMN riboswitch with equipotent activity. MIC and time-kill studies demonstrated that 5FDQD has potent and rapidly bactericidal activity against Clostridium difficile. In C57BL/6 mice, 5FDQD completely prevented the onset of lethal antibiotic-induced C. difficile infection (CDI). Against a panel of bacteria representative of healthy bowel flora, the antibacterial selectivity of 5FDQD was superior to currently marketed CDI therapeutics, with very little activity against representative strains from the Bacteroides, Lactobacillus, Bifidobacterium, Actinomyces, and Prevotella genera. Accordingly, a single oral dose of 5FDQD caused less alteration of culturable cecal flora in mice than the comparators. Collectively, these data suggest that 5FDQD or closely related analogs could potentially provide a high rate of CDI cure with a low likelihood of infection recurrence. Future studies will seek to assess the role of FMN riboswitch binding to the mechanism of 5FDQD antibacterial action. In aggregate, our results indicate that riboswitch-binding antibacterial compounds can be discovered and optimized to exhibit activity profiles that merit preclinical and clinical development as potential antibacterial therapeutic agents."

"...Treatment with a single 10-mg/kg or 5- mg/kg oral dose of 5FDQD did not affect the visible morphology of the ceca and had very little effect on the densities of the cultured cecal flora groups compared to the vehicle-treated mice. The only difference observed was a 5-fold increase in the density of members of the family Enterobacteriaceae. Thus, even at a dose 5-fold above the efficacious dose, 5FDQD did not dramatically alter the cecal morphology or flora. Speculation on why 5FDQD is highly selective is presented in Discussion (see below)."

"...In its current form, 5FDQD is a potent C. difficile antibacterial agent with an MIC90 of 1 μg/ml among strains tested herein, including four hypervirulent NAP-1 isolates. The C. difficile MIC90 values for 5FDQD are equivalent to those reported for metronidazole, vancomycin, and surotomycin, a phase III investigational agent for CDI therapy (56, 69). Although 5FDQD is less potent against C. difficile than fidaxomicin is, the available cecal concentration of 5FDQD after a single oral dose is well above the MIC and at a concentration that rapidly kills C. difficile in vitro. Accordingly, 5FDQD can completely prevent the onset of severe antibiotic-induced CDI in mice at an oral dose of 10 mg/kg twice daily for 5 days. In three repeated side-by-side comparisons, 10 mg/kg 5FDQD displayed potency equivalent to that of 3 mg/kg fidaxomicin. Given the high cecal bioavailability after dosing, it is likely that lower doses would also be effective. Future studies could be designed to determine a minimal effective dose of 5FDQD. However, it might also be beneficial to determine the maximal tolerated dose and whether higher concentrations of drug might clear an infection sooner and with fewer doses. Overall, the antibacterial activity and in vivo efficacy of 5FDQD against C. difficile are comparable to those of known CDI therapeutics or development candidates."


This recent press release contains a similar claim backing up the study above. For some reason the article is only available through Google's cache but it should be readable.
Link to causality: Industry needs to better prove pre and probiotic effects

"...Beudeker made reference to DSM’s research in the probiotic abilities of riboflavin and its multifaceted role in the human intestine. "We have recently been able to demonstrate the abilities of riboflavin as an electron acceptor as used by microorganisms contained in faecal bacteria. Just changing the dosage of riboflavin, you can change the composition of the microbiome in the colon. We now need to make that link between those results and the health benefits.""

 

[Sucrates]

http://sci-hub.cc/10.1038/ejcn.2016.119
The prebiotic concept and human health: a changing landscape with riboflavin as a novel prebiotic candidate?

"The effect of riboflavin on F. prausnitzii may be of clinical interest because, in contrast to bifidobacteria and lactobacilli, F. prausnitzii is a bacterium that directly produces butyrate.90 In addition, F. prausnitzii has been shown to possess anti-inflammatory properties and improve gut barrier function, not only through the production of butyrate91 but also by producing specific anti- inflammatory peptides.56,92 Interestingly, IBD and especially Crohn’s disease are characterized by low levels of F. prausnitzii and an increased number of E. coli and other Enterobacteriaceae. This ratio of F. prausnitzii/E. coli may be indicative of oxidative stress during gut inflammation.93 Neutrophils and other immune cells may cause an oxidative burst of reactive oxygen species that raise the redox potential to an oxidative state.94–96 An improve- ment in the ratio of F. prausnitzii/E. coli in favor of F. prausnitzii may, therefore, indicate an improvement in redox conditions, which may be beneficial to restore dysbiosis during a remission period of IBD. Whether the use of riboflavin is a therapeutic or a supportive agent during the treatment of IBD, and especially Crohn’s disease, or in healthy subjects is currently unknown but warrants urgent investigation."

 

[haidut]

http://sci-hub.cc/10.1038/ejcn.2016.119
The prebiotic concept and human health: a changing landscape with riboflavin as a novel prebiotic candidate?

"The effect of riboflavin on F. prausnitzii may be of clinical interest because, in contrast to bifidobacteria and lactobacilli, F. prausnitzii is a bacterium that directly produces butyrate.90 In addition, F. prausnitzii has been shown to possess anti-inflammatory properties and improve gut barrier function, not only through the production of butyrate91 but also by producing specific anti- inflammatory peptides.56,92 Interestingly, IBD and especially Crohn’s disease are characterized by low levels of F. prausnitzii and an increased number of E. coli and other Enterobacteriaceae. This ratio of F. prausnitzii/E. coli may be indicative of oxidative stress during gut inflammation.93 Neutrophils and other immune cells may cause an oxidative burst of reactive oxygen species that raise the redox potential to an oxidative state.94–96 An improve- ment in the ratio of F. prausnitzii/E. coli in favor of F. prausnitzii may, therefore, indicate an improvement in redox conditions, which may be beneficial to restore dysbiosis during a remission period of IBD. Whether the use of riboflavin is a therapeutic or a supportive agent during the treatment of IBD, and especially Crohn’s disease, or in healthy subjects is currently unknown but warrants urgent investigation."

Thanks for the link.

 

[Tarmander]

I like B2 more and more, but the insomnia effect of it always seems to creep up on me.

 

[nograde]

The hyperlipid guy, who eats mostly butter, explained that the B-Vitamins act as messenger molecules for your intestinal bacteria and influence biofilm formation. Ingesting them at high doses is always like playing russian roulette. For me folate is the worst, be it as very pure supplement or even liver or high amounts of OJ turns things downhill very fast. I strictly do transdermal supplementation only for that reason.

 

[Wilfrid]

Some clinical trials are actually made to evaluate the use of B2 as a novel therapeutic agent for crohn's disease.
However, I think that, if there is indeed beneficial effect of B2 on the disease outcome by stimulation of F. prausnitzii endogenous production, it would at least highly depends on the patient's age.
In pediatric crohn's disease, the inverse correlation is true, high level of F. prausnitzii are found in inflammed/diseased intestine.
As for the n=1 experience, I always reacted very badly to B2 supplementation.
Like nograde, i'm on the cautious side with those therapeutic approaches.

 

[DaveFoster]

@haidut Do you know why R5P products have such a high degree of impurity? Is it bacterial products that Peat talks about? Here's an example of a COA from PureBulk's R5P.

 

[haidut]

@haidut Do you know why R5P products have such a high degree of impurity? Is it bacterial products that Peat talks about? Here's an example of a COA from PureBulk's R5P.

Well, that's just one vendor. A purity that low is probably very cheap to obtain. I have seen much higher purity products from big vendors like Sigma. Not saying anything bad about PureBulk, just that there is higher purity available but it is MUCH more expensive.

 

[DaveFoster]

Well, that's just one vendor. A purity that low is probably very cheap to obtain. I have seen much higher purity products from big vendors like Sigma. Not saying anything bad about PureBulk, just that there is higher purity available but it is MUCH more expensive.

Thanks for the info; I checked out Sigma Aldrich's supply:

http://www.sigmaaldrich.com/catalog/DataSheetPage.do?brandKey=SIAL&symbol=R7774

They have the same assay range (industry standard), but I'd need to request a specific COA. That 1.3% might have some junk in it from PureBulk; although they're an affordable retailer, they can't compete, you're right. I'll post the reply from their e-mail when I get it.

 

[Broken man]

How much B2 I need to try this experiment please? Thank you.

 

[Lucenzo01]

How much B2 I need to try this experiment please? Thank you.

"The HED was just 1mg/kg daily and the treatment lasted for 5 days, but even a single dose worked very well." From the first post. Riboflavin is great.

 

[Broken man]

"The HED was just 1mg/kg daily and the treatment lasted for 5 days, but even a single dose worked very well." From the first post. Riboflavin is great.

Thank you so much, I want try it for my digestion.

 

[Waynish]

So we know its better to have during the day, due to effects on circadian rhythm. But any signs showing if having a lower dose spread throughout the day is more effective than a larger dose around mid-day, or vice versa?

 

[Amazoniac]

Nicotinamide, NAD(P)(H), and Methyl-Group Homeostasis Evolved and Became a Determinant of Ageing Diseases: Hypotheses and Lessons from Pellagra

"Flow of energy largely determines the structure of ecosystems via food webs and taste sensors with appetite and quality controls, that avoid toxins but allows self-medication, and involve ATP sensitive autophagy in hypothalamic neurones [97–99]. Diet with concurrent domestication of microbes as gut symbionts (or yeasts to ferment foodstuffs) that break down proteins or indigestible starches or manufacture vitamins, including nicotinamide, has evolved to supply and translocate energy and information via circadian pulses of NAD(H): particular symbionts, for example, buffer particular diets such as those low in meat and dairy consumption or high in plant based carbohydrates [100, 101]. Disturbance of these symbiotic relationships, such as by the over-use of antibiotics, is receiving considerable attention in several conditions that are increasing in incidence currently [102]. Swopping production of vitamins and some catabolic enzymes from personal metabolism to diet or symbionts appears to help find hydrogen-based energy [103]. Diets (that both depend on and have shaped cultural niche constructions, such as farming selected crops, domestication of animals for milk and meat, and cooking) and the entangled symbionts which form the microbiome in the gut are essential synergies for the evolution of human life and energy harvesting directly through NADH/ATP production [104, 105]. Diet and symbionts compensate for each other by acting as coupled recycling systems at many metabolic, immunologic, and behavioural levels, and both are implicated when energy balances go wrong as in the case of obesity, and when both are insufficient, gut cells may even break down their own components to obtain energy [106–111].

Failure of the supply of NAD(H) as a consequence of malnutrition or starvation triggers other attempts to compensate by autocarnivory to avoid apoptosis or necrotic cell death, particularly important for neurones and other cells that are rarely renewed after birth [112, 113]. Immune/inflammasome modulation with such malnutrition also occurs and allows the organism to tolerate microorganisms which are potentially deleterious but are also capable of supplying NAD(H) or its precursors [114]. Reversion to fermentation occurs in inflamed or cancerous tissue, favouring net NADH export (the Warburg phenomenon) and interacts with autophagic mechanisms with recruitment of somatic oncogene mutations and methyl-group sensitive epimutations to achieve this metabolic milieu [115–119]. All the above may be homeostatic short-term attempts to correct a poor local energy/redox environment whether by producing, redistributing, or wasting energy to heat."

"Both AIDS sufferers and pellagrins get many parasitic infections which may be dangerous homeostatic responses that boost NAD(H) in the host when dietary sources of energy are poor. Tuberculosis, for instance, excretes nicotinamide, is inhibited by nicotinamide and related compounds such as Isoniazid and treatment can precipitate pellagra and many gut infections/parasites break down otherwise indigestible cellulose, ultimately producing NAD(H) [184]. The HIV disease marker CD38 is a NADase that regulates intracellular NAD(H), implying disturbed nicotinamide-NAD(H) metabolism [185]."

"enough NAD from vitamin B3 sources means less toleration of symbionts and at the extreme immune intolerance and allergic disorders [194, 195]."
He's proposing that we're now facing a time of abundance of nutrients, and therefore less dependence on microbes and as a consequence more conflict. It's similar to burtlan that is so self-sufficient and desired that he has to walk with a baseball bat on the streets because the girls just can't accept a "No, I'm not interested".

"In the case of the so-called “infectious chronic unconquerables”, such as malaria and TB, there appears to be an immunological stand-off in which a degree of tolerance may have been traded for an increased supply of an essential nutrient. Many apparently healthy carriers, which were present in the dietary deficient population, literally disappeared as dietary standards improved in Europe and the USA. Thus, with the increasing quality of diet, symbionts become no longer necessary to supplement diet and the immune system can revert from controlling the population size of a mutualist parasite to trying to kill every invader. Recent information on the role of NAD modulating innate immunity suggests complex energy-centred interactions between neuronal and immunological systems. Contextually useful organisms (as seen in cases of pellagra) depend on ecological factors such as diet rather than geography or phylogenetic considerations, and on a good diet with changed prebiotic composition may no longer be advantageous. With the improved western diet, the immunological reaction to the formerly “useful” and actively welcomed biotic antigens concerned may be “confused” and may extend through molecular mimicry and nutrition-related signals to other antigens, thus setting off abnormal inflammatory and autoimmune reactions including to foods and pollens (that contain NAD(P)H oxidases). This is a different explanation to the more restricted hygiene hypothesis where lack of exposure rather than actively shunning “old friends” is postulated to cause immunological hyperreactivity [247–251]."

"Placebo effects that are so prominent in man also have been linked to energy pathways and are particularly effective when the energy future is perceived as healthy, allowing energy to be invested in healing rather than fire fighting [305]."​

 

[GAF]

I have taken 250 to 900 mg B2 daily for 3 1/2 months. I have taken it right before bed on several occasions, usually after a evening of excess dancing fun. Out of maybe 10 times, twice I have had trouble getting to sleep. I can't say whether the B2 was a contributor or if it was I took a good long nap before I went out.

B2 has not made my pooping perfect.

I have not been ill, gotten a cold, or anything since experiment began.

Now I am taking 400 mg in the morning usually but sometimes early afternoon, and sometimes both.

B2 affects a person head to toe. My results have been spectacular. Nothing else I have ever tried is even measurable in comparison.

 

[dfspcc20]

I have taken 250 to 900 mg B2 daily for 3 1/2 months. I have taken it right before bed on several occasions, usually after a evening of excess dancing fun. Out of maybe 10 times, twice I have had trouble getting to sleep. I can't say whether the B2 was a contributor or if it was I took a good long nap before I went out.

B2 has not made my pooping perfect.

I have not been ill, gotten a cold, or anything since experiment began.

Now I am taking 400 mg in the morning usually but sometimes early afternoon, and sometimes both.

B2 affects a person head to toe. My results have been spectacular. Nothing else I have ever tried is even measurable in comparison.

Do you take any other B vitamins?

 

[Mossy]

I have taken 250 to 900 mg B2 daily for 3 1/2 months. I have taken it right before bed on several occasions, usually after a evening of excess dancing fun. Out of maybe 10 times, twice I have had trouble getting to sleep. I can't say whether the B2 was a contributor or if it was I took a good long nap before I went out.

B2 has not made my pooping perfect.

I have not been ill, gotten a cold, or anything since experiment began.

Now I am taking 400 mg in the morning usually but sometimes early afternoon, and sometimes both.

B2 affects a person head to toe. My results have been spectacular. Nothing else I have ever tried is even measurable in comparison.

If it helps, I can tell you for certain B2 effects my sleep. Before I read the end of your sentence, and read "I have taken it right before bed...", the first thing I though was wow, and it didn't' affect his sleep? But, I see that you did have some trouble part of the time. It does give life to my body, but the poor sleep burns me out in the end. When I have the courage, I am going to do this 5 day regimen, in the hopes it will really revitalize my gut. I say courage, because if I take 82 mg/day for 5 days, I'll be replicating da Vinci's Sistine Chapel ceiling on my bedroom ceiling, will all the energy and focus I'll have.

 

[GAF]

Yes, B1 500mg and B6 maybe weekly and an occasional B complex/energin. For my experiment, I have been a B2 bomber.

My point is that B2 is the easiest, cheapest and fastest way to increase energy and focus, if one is lacking. My 84 yr old parents will agree.

Try this, next time you guys get in the car to take a 5 hour drive, down a big B2. You still have to eat and drink...

 

[Lucenzo01]

"Flow of energy largely determines the structure of ecosystems via food webs and taste sensors with appetite and quality controls, that avoid toxins but allows self-medication, and involve ATP sensitive autophagy in hypothalamic neurones [97–99]. Diet with concurrent domestication of microbes as gut symbionts (or yeasts to ferment foodstuffs) that break down proteins or indigestible starches or manufacture vitamins, including nicotinamide, has evolved to supply and translocate energy and information via circadian pulses of NAD(H): particular symbionts, for example, buffer particular diets such as those low in meat and dairy consumption or high in plant based carbohydrates [100, 101]. Disturbance of these symbiotic relationships, such as by the over-use of antibiotics, is receiving considerable attention in several conditions that are increasing in incidence currently [102]. Swopping production of vitamins and some catabolic enzymes from personal metabolism to diet or symbionts appears to help find hydrogen-based energy [103]. Diets (that both depend on and have shaped cultural niche constructions, such as farming selected crops, domestication of animals for milk and meat, and cooking) and the entangled symbionts which form the microbiome in the gut are essential synergies for the evolution of human life and energy harvesting directly through NADH/ATP production [104, 105]. Diet and symbionts compensate for each other by acting as coupled recycling systems at many metabolic, immunologic, and behavioural levels, and both are implicated when energy balances go wrong as in the case of obesity, and when both are insufficient, gut cells may even break down their own components to obtain energy [106–111].

Failure of the supply of NAD(H) as a consequence of malnutrition or starvation triggers other attempts to compensate by autocarnivory to avoid apoptosis or necrotic cell death, particularly important for neurones and other cells that are rarely renewed after birth [112, 113]. Immune/inflammasome modulation with such malnutrition also occurs and allows the organism to tolerate microorganisms which are potentially deleterious but are also capable of supplying NAD(H) or its precursors [114]. Reversion to fermentation occurs in inflamed or cancerous tissue, favouring net NADH export (the Warburg phenomenon) and interacts with autophagic mechanisms with recruitment of somatic oncogene mutations and methyl-group sensitive epimutations to achieve this metabolic milieu [115–119]. All the above may be homeostatic short-term attempts to correct a poor local energy/redox environment whether by producing, redistributing, or wasting energy to heat."

"Both AIDS sufferers and pellagrins get many parasitic infections which may be dangerous homeostatic responses that boost NAD(H) in the host when dietary sources of energy are poor. Tuberculosis, for instance, excretes nicotinamide, is inhibited by nicotinamide and related compounds such as Isoniazid and treatment can precipitate pellagra and many gut infections/parasites break down otherwise indigestible cellulose, ultimately producing NAD(H) [184]. The HIV disease marker CD38 is a NADase that regulates intracellular NAD(H), implying disturbed nicotinamide-NAD(H) metabolism [185]."

"enough NAD from vitamin B3 sources means less toleration of symbionts and at the extreme immune intolerance and allergic disorders [194, 195]."
He's proposing that we're now facing a time of abundance of nutrients, and therefore less dependence on microbes and as a consequence more conflict. It's similar to burtlan that is so self-sufficient and desired that he has to walk with a baseball bat on the streets because the girls just can't accept a "No, I'm not interested".

"In the case of the so-called “infectious chronic unconquerables”, such as malaria and TB, there appears to be an immunological stand-off in which a degree of tolerance may have been traded for an increased supply of an essential nutrient. Many apparently healthy carriers, which were present in the dietary deficient population, literally disappeared as dietary standards improved in Europe and the USA. Thus, with the increasing quality of diet, symbionts become no longer necessary to supplement diet and the immune system can revert from controlling the population size of a mutualist parasite to trying to kill every invader. Recent information on the role of NAD modulating innate immunity suggests complex energy-centred interactions between neuronal and immunological systems. Contextually useful organisms (as seen in cases of pellagra) depend on ecological factors such as diet rather than geography or phylogenetic considerations, and on a good diet with changed prebiotic composition may no longer be advantageous. With the improved western diet, the immunological reaction to the formerly “useful” and actively welcomed biotic antigens concerned may be “confused” and may extend through molecular mimicry and nutrition-related signals to other antigens, thus setting off abnormal inflammatory and autoimmune reactions including to foods and pollens (that contain NAD(P)H oxidases). This is a different explanation to the more restricted hygiene hypothesis where lack of exposure rather than actively shunning “old friends” is postulated to cause immunological hyperreactivity [247–251]."

"Placebo effects that are so prominent in man also have been linked to energy pathways and are particularly effective when the energy future is perceived as healthy, allowing energy to be invested in healing rather than fire fighting [305]."​

Extremey interesting article. You should made a thread with it.

 

[Antonello]

@haidut For some reason this study doesn't work for me, when I take riboflavin it mess up my digestion and I get painful diarrhea and gas, it's like it feeds intestinal bacteria but the bad one because once it stop the diarrhea if feel awful for a few days. I was reading about b vitamins soluble in water and etanol like your energin. do you think topical application can avoid the digestion but still have a good effect on bacteria?
do you need etanol for topical absorption or a simple bath with a pinch of riboflavin will to the trick?