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Wednesday, 8 March 2017

Take your Bumetanide Studies with a Pinch of Salt



This blog does try to be based on evidence, but sometimes you do have to question the validity of what appears in peer reviewed journals.  It might concern what does, or does not cross the blood brain barrier, or what works in vivo versus in vitro.

Two interesting papers were recently brought to my attention regarding Bumetanide.


With a pinch of salt is an English idiom which means
to view something with skepticism 



In Tyler’s paper it was rats with epilepsy showing big improvements when taking Bumetanide. 

In Agnieszka’s paper, involving mice and Chinese hamsters, researchers are making the point that so little Bumetanide crosses into the brain that its therapeutic value is limited. 

So which is true? 

Well it seems that in some humans with autism enough bumetanide crosses the blood brain barrier (BBB) to show a positive effect.  Perhaps if a more penetrative analogue of Bumetanide was developed, it would show even greater effect, otherwise adjunct therapies may be needed (Acetazolamide, potassium bromide, estradiol etc) to gain the full benefit of lowering intracellular chloride. 

In the past I have made the case for bumetanide possibly reducing the incidence of epilepsy developing in autism and that this really would be important. This does not mean that one person with autism might not develop epilepsy around the same time he started taking bumetanide. In the study below the rats with seizures seemed to be protected by bumetanide and the number of harmful neural connections detected in the brain was significantly reduced. 




Abstract

There is accumulating evidence that bumetanide, which has been used over decades as a potent loop diuretic, also exerts effects on brain disorders, including autism, neonatal seizures, and epilepsy, which are not related to its effects on the kidney but rather mediated by inhibition of the neuronal Na-K-Cl cotransporter isoform NKCC1. However, following systemic administration, brain levels of bumetanide are typically below those needed to inhibit NKCC1, which critically limits its clinical use for treating brain disorders. Recently, active efflux transport at the blood-brain barrier (BBB) has been suggested as a process involved in the low brain:plasma ratio of bumetanide, but it is presently not clear which transporters are involved. Understanding the processes explaining the poor brain penetration of bumetanide is needed for developing strategies to improve the brain delivery of this drug. In the present study, we administered probenecid and more selective inhibitors of active transport carriers at the BBB directly into the brain of mice to minimize the contribution of peripheral effects on the brain penetration of bumetanide. Furthermore, in vitro experiments with mouse organic anion transporter 3 (Oat3)-overexpressing Chinese hamster ovary cells were performed to study the interaction of bumetanide, bumetanide derivatives, and several known inhibitors of Oats on Oat3-mediated transport. The in vivo experiments demonstrated that the uptake and efflux of bumetanide at the BBB is much more complex than previously thought. It seems that both restricted passive diffusion and active efflux transport, mediated by Oat3 but also organic anion-transporting polypeptide (Oatp) Oatp1a4 and multidrug resistance protein 4 explain the extremely low brain concentrations that are achieved after systemic administration of bumetanide, limiting the use of this drug for targeting abnormal expression of neuronal NKCC1 in brain diseases.
  

Prolonged epileptic seizures may cause serious problems that will continue for the rest of a patient's life. As a result of a seizure, neural connections of the brain may be rewired in an incorrect way. This may result in seizures that are difficult to control with medication. Mechanisms underlying this phenomenon are not entirely known, which makes current therapies ineffective in some patients.
A study conducted with a rat epilepsy model at the Neuroscience Center of the University of Helsinki showed that a change in the function of gamma-aminobutyric acid (GABA), a main neurotransmitter in the brain, is an underlying cause in the creation of harmful neural connections.
After a prolonged convulsive seizure, instead of the usual inhibitory effect of the transmitter, GABA accelerates brain activity. This, in turn, creates new, harmful neural connections, says Research Director Claudio Rivera.
The accelerating effect of GABA was blocked for three days with a drug called bumetanide given soon after a seizure. Two months after the seizure, the number of harmful connections detected in the brain was significantly lower.
"Most importantly, the number of convulsive seizures diminished markedly," says Claudio Rivera.
In this study, new indications may be found for bumetanide in the treatment of epilepsy. Bumetanide is a diuretic already in clinical use. Extensive clinical studies have already been conducted with bumetanide regarding its ability to reduce the amount of convulsions or prevent them entirely in the acute phase of seizures. This, however, is the first time that bumetanide has been found to have a long-term effect on the neural network structure of the brain.
Further study of the newly found mechanism may eventually help limit the exacerbation of epilepsy and prevent the onset of permanent epilepsy after an individual serious seizure. It may also be possible that a similar mechanism is responsible for the onset of epilepsy after a traumatic brain injury.
"The next step is to study bumetanide both by itself and in combination with other clinically used drugs. We want to find out the ways in which it may offer additional benefits in the treatment of epilepsy in combination with and even in place of currently used antiepileptic drugs," says Claudio Rivera.



Vitamin D and Autism

Two medical readers of this blog highlighted this recent paper showing an apparent universal benefit of vitamin D supplementation in autism.

Is it too good to be true?  Time for the pinch of salt?

One important point to note is that this study was in Egypt and, although sunny, are children there eating food that has already been fortified with vitamin D, like it is in Western countries?

Randomized controlled trial of vitamin D supplementation in children with autism spectrum disorder

Abstract

BACKGROUND:



Autism spectrum disorder (ASD) is a frequent developmental disorder characterized by pervasive deficits in social interaction, impairment in verbal and nonverbal communication, and stereotyped patterns of interests and activities. It has been previously reported that there is vitamin D deficiency in autistic children; however, there is a lack of randomized controlled trials of vitamin D supplementation in ASD children.

METHODS:



This study is a double-blinded, randomized clinical trial (RCT) that was conducted on 109 children with ASD (85 boys and 24 girls; aged 3-10 years). The aim of this study was to assess the effects of vitamin D supplementation on the core symptoms of autism in children. ASD patients were randomized to receive vitamin D3 or placebo for 4 months. The serum levels of 25-hydroxycholecalciferol (25 (OH)D) were measured at the beginning and at the end of the study. The autism severity and social maturity of the children were assessed by the Childhood Autism Rating Scale (CARS), Aberrant Behavior Checklist (ABC), Social Responsiveness Scale (SRS), and the Autism Treatment Evaluation Checklist (ATEC).

RESULTS:



Supplementation of vitamin D was well tolerated by the ASD children. The daily doses used in the therapy group was 300 IU vitamin D3/kg/day, not to exceed 5,000 IU/day. The autism symptoms of the children improved significantly, following 4-month vitamin D3 supplementation, but not in the placebo group. This study demonstrates the efficacy and tolerability of high doses of vitamin D3 in children with ASD.

CONCLUSIONS:



This study is the first double-blinded RCT proving the efficacy of vitamin D3 in ASD patients. Depending on the parameters measured in the study, oral vitamin D supplementation may safely improve signs and symptoms of ASD and could be recommended for children with ASD. At this stage, this study is a single RCT with a small number of patients, and a great deal of additional wide-scale studies are needed to critically validate the efficacy of vitamin D in ASD.

Conclusion
Take your research with a pinch of salt.




47 comments:

  1. Well in the study on Bumetanide not crossing the BBB, the nature of how it works is under major scrutiny these days as the BBB seems to be more or less permeable under certain non-pathological conditions and under inflammatory stress (such as in autism) it tends to be more permeable. So in a healthy brain with no inflammatory stress, maybe less Bumetanide makes it into the brain (if you assume the Chinese hamster study to be correct and apply to humans), while in a brain with increased BBB permeability due to reasons such as oxidative stress, you might get more Bumetanide. Also, ventral regions of the brain such as the amygdala and hippocampus are more susceptible to BBB permeability from oxidative stress and also happen to be major subcortical areas that seem to be very dysfunctional in autism. There is also the hypothalamus of which one of its nuclei is not really shielded by the BBB at all so that it can sample the blood for hormones, glucose, salt, and other substances that typically don't pass the BBB at all or else without some sort of regulation.

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  2. I finally got around to reading a paper I had in the queue for a few days on NAFLD (Non-Alcoholic Fatty Liver Disease) but I feel it is relevant to autism because obesity tends to track with autism and abdominal obesity tends to track with NAFLD. In fact, I think Petra mentioned it being one of the biggest comorbid concerns with her child and antipsychotics tend to promote severe weight gain as well:

    http://msb.embopress.org/content/13/3/916

    To summarize the findings, what was first found (in rats) was that there were two main factors causing hepatic steatosis and those were Glutathione exhaustion and NAD+ exhaustion. They then hypothesized that providing the precursors to NAD+ synthesis and GSH synthesis would help improve the symptoms of NAFLD in mice so they decided on a cocktail of N-Acetyl-Cysteine (NAC), Nicotanimide Riboside (NR), and Serine which is a precursor to Glycine which is necessary for recycling GSH in the liver and administered it to mice fed a western-mimicking diet (high sugar and fat) which leads to NAFLD and it largely prevented NAFLD. They then tested serine supplementation in humans with NAFLD and it significantly improved liver markers.

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    1. D-serine (DSR) has been mentioned earlier in this blog. It has been shown to help neurological disorders, particularly schizophrenia.

      "A first generation of proof-of-concept animal and clinical studies suggest beneficial DSR effects in treatment-refractory schizophrenia, movement, depression, and anxiety disorders and for the improvement of cognitive performance"

      https://www.hindawi.com/archive/2014/859735/

      When people check liver enzymes (ALT and AST)in autism, very often the results are odd.

      I think the liver is reflecting what is happening in the brain. The liver is providing a convenient biomarker. So Petra's son could get a double benefit from D-Serine.

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    2. J Agric Food Chem. 2009 Jul 8;57(13):5982-6. doi: 10.1021/jf900470c.
      Acetic acid upregulates the expression of genes for fatty acid oxidation enzymes in liver to suppress body fat accumulation.

      I have already mentioned that moderate ethanol consumption seems to hepl my son's brain process as if he were neurotypical. Ethanol is converted to acetic acid and thought there may be something there relevant to his brain/liver metabolism disorder, particularly in view of such low LDL cholesterol levels in blood and fat deposits in liver.
      If Peter, Tyler, AJ, or anyone here that could comment on this hypothesis, would be really helpful. I know alcohol is a taboo issue here, as most people are trying to treat children, but I am raising this issue just in case there are interventions that could mimic the effect of alcohol.

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    3. Petra, on a slightly different subject, is your son using verapamil?

      Calcium channel blockers as potential therapeutics for obesity-associated autophagy defects and fatty liver pathologies

      https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4502701/

      "Verapamil also reduces hepatic lipid droplet accumulation, insulin resistance and steatohepatitis, suggesting that calcium channel blockers can be used for correction of general NAFLD pathologies."

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    4. Peter, when my son had very high liver enzymes and CRP doctors told me to stop all medication in case the infection was drug induced, so I stopped Verapamil.
      Also he sometimes reacts badly even with low dose magnesium which is a calcium channel blocker.
      This extract is from your recent post:
      Calcium channel blocking drugs used to treat high blood pressure, abnormal heart rhythm, angina pectoris, panic attacks, manic depression and migraine may occasionally cause drug-induced parkinsonism.
      Such things made me think twice about Verapamil.

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    5. Petra, maybe do a trial of Verapamil and then measure the liver enzymes and see if they improve?

      The question is whether the NAFLD is a symptom of an underlying autism-related dysfunction, or just a coincidence.

      Lack of autophagy, oxidative stress and calcium channel dysfunction are features of both autism and NAFLD.

      Almost all drugs have side effects in someone, so you do have to be careful. In theory NAFLD is not treatable, perhaps because it can be caused by different things.

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    6. Ethanol does so many different things, I would not know where to start. The other question with regards to ethanol is how much. I used to ferment kefir (and no ferment Biogaia) every day and depending on how long you let it ferment, you would get different levels of alcohol. On average the kefir I would give my kids was probably 2% alcohol and I didn't really worry about such a small amount because it is not like they drank kefir by the gallon full. Very small amounts of alcohol are easily handled by the body, as are other toxins, but once you go beyond grog levels of alcohol and get into the beer/wine range, then you start getting increasingly negative health and cognitive effects from its use.

      As for drugs that mimic the effects of alcohol, well there are plenty though of course every drug is a little different from another in terms of its full spectrum of effects.

      I am pretty sure alcohol does pretty much nothing positive and only negative for the liver, contrary to what you may read in the press about red wine (the most recent studies controlled for income as wealthier people drink more alcohol because of its association with status and once that was taken into account, there were only negative health benefits from red wine).

      But if you son has cognitive benefits from alcohol, it is likely from its pro GABAergic effects and anti NMDA effects.

      There are of course other possibilities, especially from alcohol's metabolites. Acetylaldehyde causes a histamine reaction (you can get the same result from regular old niacin as well). Acetylaldehyde also has dopaminergic effects. I have read that NAC is good at countering some of the secondary health issues to alcohol exposure and I have read anecdotal resports Nicotanimide Riboside (NR) is good at increasing alcohol tolerance (i.e. preventing drunkeness), likely because alcohol reduces the NAD+/NADH ratio and NR increases the ratio (this is speculation of course).

      One way to give you a hint about whether it is the alcohol itself or the metabolites and secondary effects that are causing these cognitive improvements would be to take NAC and NR a half hour before ingesting alcohol and see if the positive cognitive changes you speak of are not blunted or removed altogether. If they are not, then it is likely the GABAergic effects, and if they are blunted, then it may be dopaminergic from the acetylaldehyde.

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  3. Peter, I would like to trial D-serine for my son.
    It lowers cortisol and lets glucose be utilized by cells, among other possible benefits.
    There is a product from Now Foods 100mg and Solgar 500mg, both from soya. I read that soya may not be as good as bovine sources but then there is always the risk of mad cow disease.
    If you eat 100mg chicken hearts you can get 400mg D-serine in theory.
    What do you recommend?

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    Replies
    1. Petra, use google to look at clinical trials of D-serine in schizophrenia. The doses seem to be 30mg/kg and higher. I would use a standardized dose so you can make a rational trial. Soja based supplements look best. You can probably find feedback from current users via google.

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  4. Tyler, I was wondering if D.serine would be a good choice for my son, what do you think? have you tryed it?
    Valentina

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    Replies
    1. No I have not tried it. Also, in the study I alluded to I believe they used L-Serine which is much different than D-Serine in terms of its effects in the body and brain.

      Delete
    2. Tyler, my son´s godfather arrives next week, I added one more jar of NR and L-Serine from bulk.
      Valentina

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  5. Petra, I think phosphatidylserine would be a better choice than d serine alone and mixed with PE, phosphatidyl ethanolamine and PC,phosphatidylcholine. You can find this mix in amazon, soy free.The brand is Natural Bioscience
    Valentina

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  6. Peter, My son's seizures increased in frequency as I was weaning from breastfeeding. I wonder if the hormones in the milk might have had some effect looking back. I believe the hormones had a neuroprotective effect. Estradiol, progesterone, prolactin, thyroxine, triiodothyronine, cortisol, prolactin, EGF, Thyroid hormones, Relaxin, Beta Endorphins, EPO, GnRH, Insulin, androgens, gastrin, adiponectin, resistin, ghrelin, and leptin......Then they get AA's, nutrients, living cells, and enzymes. Looking at your interesting diagrams I can see how certain hormones might help.

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    Replies
    1. Hormones act in surprisingly connected ways and in autism some are clearly disturbed. It is just not easy to now which one(s) could be modified for a long-lasting positive effect. I think a little extra estradiol may do the trick for some. It will reduce testosterone and I think leptin. Leptin is the subject of a new post and is increased in early onset autism and will contribute to inflammatory-type symptoms. RORalpha is linked to the relationship between testosterone and estrogens. RORalpha4 affects the leptin produced in fat cells. So extra estradiol will affect RORalpha and indirectly hopefully also reduce leptin production.
      RORalpha appears to be a hub where things going wrong in autism can be traced back to.

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  7. Petra, my husband was found to have mild fatty liver a few years back. At that time, I read that phosphatidylcholine was the best for it. He used it for several months. Since then, he has made sure that he eats at least two eggs every day, for the choline. Maybe you can run a small trial and test.

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    Replies
    1. RG- Make sure that the egg yolks aren't overly cooked as this reduces the choline available and requires more antioxidants to process in the body.

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  8. Hi dear all,

    I'm the 1st author of the paper mentioning great improvement of bumetanide on harmful neuronal network connections and subsequent seizures in rats. I'm a PhD in neuroscience, not a MD, so I may not have hints for the patients but could share my modest knowledge on the field. First of all, thank you for lifting up this very interesting topic.
    As concerning the paper demonstrating that bumetanide poorly cross the BBB, this is not the sole and only respectful study showing that and the paper depicted in this blogspot comes from a well-known, respected, talented and real specialist, Dr. W. Loscher who has unequivocally showed for decades the limitations of bumetanide on crossing the BBB. In our paper (beneficial effect of bumetanide on seizure), we in purpose used all the knowledge gained in part by Loscher Lab and others and used bumetanide not iv or ip but bumetanide was applied directly into the brain using osmotic mini-pumps. This to circumvent on the known limitation of this drug in crossing the BBB. I agree with the 1st comment of Tyler claiming that BBB can be disrupted in many cases such as inflammation, traumatic brain injury and so. This said bumetanide had no effect on newborn seizures in humans and had detrimental effect on hearing (http://www.thelancet.com/journals/laneur/article/PIIS1474-4422(14)70303-5/abstract). However, although the exact mechanism of action remain unknowed, nowadays promising resultants under clinical trials show stunning outcomes on autism syndrome disorder using bumetanide in humans (http://www.neurochlore.fr/research/). In conclusion, our intent is to show bumetanide or its pro-drugs (as mentionned in the comments above, and as developed in rodent by Loscher lab) is really potent by itself on epileptic brain in vivo and could not only reduce the convulsions in adult rats but also constrain the underlying neuronal circuit supposedly at the origin of the seizures to have long-term effects. It is a fundamental research aiming at showing proof-of-concept findings rather than possibility for straightforward clinical translation from rat to human.
    But there is more hope for bumetanide since it is an FDA approved drug already available and in use... Also a case report showed direct brain delivery of valproate to dampen seizures in australia (http://www.abc.net.au/news/2016-12-15/world-first-trial-for-new-epilepsy-producing-promising-results/8122668), similarly to what we did in our paper with bumetanide in rats. Many more efforts are needed though if one day bumetanide would be used to cure adult epileptic patients...

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  9. Peter, how can I get bumetanide as it's for heart condition, it's hard getting the prescription from a psychiatrist or doctor. I feel like it's not their medicine

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    1. It depends where you live. People in the US/Canada are buying online from Mexico. In Europe it us available in Spain by asking in a pharmacy, no prescription needed.

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    2. Is this the tablet form? Here in Australia is have to get it from doctor.

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    3. It is a prescription drug. It is likely to be approved in Europe as an autism therapy in 2021. Until then it is just an off-label therapy of a drug used to lower blood pressure. It is a tablet. You would have to show the research to your doctor to persuade him to prescribe it, or buy it on line from an overseas internet pharmacy.

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  10. Hi Peter, just last year I've been diagnosed with Aspergers. I tried all possible medications since and nothing really helped me. I think Bumetanide would make a great effect on me, but I am not able to get a hand on it. My doctor would prescribe it to me immediately, if there would only be an official product in my country, which isn't the case. I looked through all of the comments in this blog and googled a lot of the online pharmacies to find a good source for bumetanide. Sadly, all the online pharmacies are rated as "fraud"/"scam" on rating sites. Thus said, I hope that you or any reader may help me to find a reliable way to buy the medicine. I am located in Central Europe. Where can I get Bumetanide? To other Readers from Europe: Where do you buy your Bumetanide? (I read someone from Romania gives it to their child)(someone said, I could just buy it in a Pharmacie in Spain, do they have it in Spain?)

    PS: maybe you should consider making a post about, how to get bumetanide safely. Not to advertise, but to make sure, that people already suffering from autism, do not suffer more by buying drugs online or getting scammed. It is still a long way to go till 2021 and many poeple will find hope in your blog. This Question will thus arise a lot more i think.

    Love your blog and all the best

    Stefan


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    Replies
    1. Stefan, if your doctor will write you a prescription, you will find that pharmacies in other European countries will accept that prescription. Legitamate UK online pharmacies should accept a prescription from another EU country and send bumetanide to you. From personal experience I can tell you that German and Italian pharmacies are very happy to accept prescriptions from foreign countries, even if you want a year's supply.

      It is only a problem if you do not have a prescription. There is a good reason for this.

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    2. Since you mentioned Spain - I know that some time ago it was available in Spain (Fordiuran) and it did not require prescription.

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    3. To some point maybe it is worth to mention that although it has a favorable benefit/risk ratio "The most frequent adverse events were hypokalemia, increased urine elimination, loss of appetite, dehydration and asthenia. Hypokalemia occurred mainly at the beginning of the treatment at 1.0 and 2.0 mg twice-daily doses and improved gradually with oral potassium supplements. The frequency and incidence of adverse event were directly correlated with the dose of bumetanide" as stated in Lemmonier et al., Effects of bumetanide on neurobehavioral function in children and adolescent with ASD Translational Psychiatry 2017. SO IT IS CRUCIAL NOT TO SELF-MEDICATE WITHOUT BEING FOLLOWED BY DOCTORS while taking bumetanide.

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  11. I felt like your blog didn't give the Vitamin D study much credence and wanted to share our experience with high-dose Vitamin D.

    We began giving 3000 IU / day to our 35lb 3 year old son in February. The effects occurred within days, much more rapidly than we had anticipated. His teachers and therapists were stunned and asked us what changed. Before had we had a very irritable child that would cry and fuss for up to a half hour at the beginning of any form of structured play, refused to imitate any words or make substantial eye contact. Even verbally labeling what he was doing would routinely cause a prolonged outburst. After adding the D/K2/A into the mix, he became much more calm, imitative, and interested in playing with us. Word usage has been slower to develop, but he now routinely echos words now and spontaneously uses limited two word phrases like "look outside" or "shoes off." Importantly, he can tolerate us talking to him and telling him words rather than just exploding whenever we talk to him.

    We went from having our son categorized as being severely mentally handicapped at 2.5 years of age (IQ estimated at 47; everything from fine motor skills to language ability assessed as severely handicapped) to an estimated IQ of 80 (to the point the interventionists say he won't require special instruction for cognitive impairment), in the course of three months. His expressive vocabulary went from 2 words to about 20 in the same time, and he started parroting several dozen words upon prompting; learned and would say all the names of his classmates in that time as well. He also became much more tolerant of loud or sudden noises, and has taken great interest in music, and generally become much more affectionate (not neurotypical level, yet, but from nothing springs something). He's learned how to play; build things, and just generally acts more normal. He still has some stimming, still does some things even after being asked repeatedly not too (e.g. throwing sand out of a sandbox or spilling water because he wants to watch it pour out of something), but I think some disobedience is normal at this age. He no longer does self-hurting behaviors as he did before, however.

    4 months after starting Vitamin D, another independent assessor (his speech therapist) has had to throw out her annual treatment plan, because he had already met all the milestones in only 3 months, and no longer fits her original assessment of moderate to severely mentally handicapped, emotionally handicapped, and speech delayed. Instead, he just has delayed speech and some mild signs of autism.

    Could it be coincidence? Sure. I know better than to say it's definitive that the D that did it. The only other variable we added to the equation was starting 2 mg daily of folinic acid about 2 weeks after the D; and this is after the teachers had noticed the behavioral improvements. I think it may have helped some with the language development but the effects are much less dramatic. Based on the Egyptian study on D, which did note a small percentage (maybe 5%) of non-responders, along with a significant percentage of very strong responders, I think it's something that virtually everyone with autism should try. Of course it's possible that he'd have just had such an amazing transformation without the D because some gene somewhere turned on, or some critical process in his neurodevelopment occurred. But given that this wasn't just one aspect of the disorder that was affected; it was many aspects, which improved in a short time, and noted by at least 3 separate therapists who were unaware of the intervention, I'm inclined to believe the D had something to do with it.

    Are such high levels safe? After doing some homework, I found that even much higher levels of D (20x more than advocated in this study) are generally well-tolerated, especially if co-administered with Vitamin A and K2, which we added to be on the safe side as well.

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    1. JG, it is great that your child has improved so much. Do you know if he was deficient in vitamin D at the start? Are you using a high dose of K2? Some people respond very well to K2.

      Many people were sceptical about the Egyptian study on vitamin D, because of how many responders there were. In most countries it is hard to avoid vitamin D because it is added to basic foods.

      I don't suppose you want to try stopping the vitamin to check the effect.

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    2. JG, there is a fairly large study planned to take place in New Zealand. This should be able to validate the Egyptian study.

      Vitamin D and omega-3 fatty acid supplements in children with autism spectrum disorder: a study protocol for a factorial randomised, double-blind, placebo-controlled trial

      https://trialsjournal.biomedcentral.com/articles/10.1186/s13063-016-1428-8

      You can have too much vitamin D and it is easy to check. Also, high amounts of vitamin D affect calcium levels in your blood. Calcium is a key signaling ion and in some people taking calcium supplements makes them much worse, but in autism usually both extremes exist. I would check 25(OH)D and calcium in his blood. Vitamin D accumulates since it is not water soluble. If 25(OH)D levels are above 150 ng/ml this is considered potentially toxic.

      https://www.vitamindcouncil.org/about-vitamin-d/am-i-getting-too-much-vitamin-d/

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    3. Regarding the vit D , my daughter whi shows some of autistic properties like routine activities, stubborn behaviour beside lowee sociable awarness than her age , she was previously diagnosed adhd but the autism charcter is more obvious , we made an analysis for vit D and the result was 16 , considered deficiency, we started the treatment and we noticed the disappearance of the certain repeated behaviour, like obssesive sounds or arm movements she used to have. In the first week of taking vit D , we should continue till 12 weeks , although the stubborn behaviour and persistance is still there strongly affceting badly her social life

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  12. Hold on with me, this is long but It seems im onto something here:

    This might be to interest to those who use bumetanide, this is a somewhat old study it seems, but relevant:

    Elevated levels of sodium blunt response to stress, study shows:
    https://www.sciencedaily.com/releases/2011/04/110405175012.htm

    As I suspected, water balance/sensors play a huge role in social behavior (and that is to be expected as it is critical for survival).

    Some highlights:

    "All those salty snacks available at the local tavern might be doing more than increasing your thirst: They could also play a role in suppressing social anxiety. New research shows that elevated levels of sodium blunt the body's natural responses to stress by inhibiting stress hormones that would otherwise be activated in stressful situations."

    "Compared with a control group, the rats that received the sodium chloride secreted fewer stress hormones and also displayed a reduced cardiovascular response to stress."

    "The elevated sodium level, known as hypernatremia, limited stress responses by suppressing the release of the pro-stress hormone angiotensin II. CONVERSELY, IT INCREASED THE ACTIVITY OF OXYTOCIN, an anti-stress hormone"

    Now I have mentioned before that I never throw salt on my food, this was not just a concious decision I just never did it!!
    I never questioned myself WHY?, but recently after reading about your bumetanide (diuretic after all) post and the effiacy in autisms it certainly raised my interested.

    On top of that me and a few others have mentioned the excessive thirst, and I have wondered myself If I subconciously was not eating salt in order to induce thirst/mild dehydration thereby somewhat mimicking the effect of bumetanide.

    I think the key to unlocking and better understanding this is to WHY I do not put salt on my food and subconsiously put myself in diuretic mode.

    There is probably something about diuresis that upregulates certain receptors in the brain as a response, the question is what and why.
    Since this study suggest that salt intake INCREASES oxytocin, this makes sense both no sense and it does make sense to me at the same time.

    Basically low water concentrations in the body would lower oxytocin, but what I have found striking about my form of autism (aspergers) is that there is mainly a SOCIAL WANTING DYSFUNCTION, for me the experience of social interaction if I like someone is intact, but only if it happens unexpectedly (through a stressor).

    It seems in autism the brain is not properly capable of 'seeing' the benefits of social interaction (shelter, food, water).
    This results in not calling friends and even not wanting affection from their parents.

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  13. There for my hypothesis is that we have a 'defense mechanism/defense adaptation' dysfunction in the brain, and that the pathways of empathy are actually intact but they just do not get activated.

    Diuresis could cause temporarily 'spark' these underactive systems, reflecting in improvements in behavior

    Social ‘WANTING’ (not social experiencing) dysfunction in autism: neurobiological underpinnings and treatment implications:
    https://jneurodevdisorders.biomedcentral.com/articles/10.1186/1866-1955-4-10



    "This is most likely caused by dysfunction of the dopaminergic–oxytocinergic ‘wanting’ circuitry, including the ventral striatum, amygdala, and ventromedial prefrontal cortex."

    ". In fact, several recent imaging studies on resting-state functional connectivity and DTI confirm disruptive neural activation
    dynamics in ASD within the vmPFC–VS–amygdala circuitry [78-81]. THESE FINDINGS ARE ALSO IN LINE WITH THE IDEA OF ASD AS A NEURFUNCTIONAL DISCONNECTION SYNDROME [82-84], most likely mediated by COMPLEX GENETIC FACTORS (for example, synaptic cell adhesion plasticity [85]), which affect efficient information transfer within the mesocorticolimbic reward circuitry and may cause aberrant motivation, that is, affect ‘wanting’ tendencies"

    Fever, dehydration/diuresis, starvation, sleep deprivation are all forms of stressors for the body and probably induce responses by TEMPORARILY upregulating/strengthening these systems.

    A pill that mimics these stressors would fix empathic dysfunction im 100% sure of it.
    I believe I have posted it before on here and certainly have on other forums, the day AFTER 12-15 beers (YES THAT IS WHAT OTHERS EXPERIENCE AS A HANGOVER), im 100% cured for like 5-8hours, also sleep deprivation fixes me up to 85% emotional capacity.

    Now alcohol is pure toxic, hell I would never ever recommend it to anyone, in fact I have alcohol aversion (as explained before in my older posts, complex, but long story short has to do with what I believe diminished protein kinase a activity).

    During a hangover loads of things happen, serotonin depletion in the amygdala, NMDAr upregulation, 5ht1a autoreceptors upregulation (this decreases serotonin!), not to mention alcohol is also a strong diuretic and powerfully lowers blood sugar.
    Alcohol also strongly increases cortisol and aspergers is associated with a decrease in cortisol awakening response.

    Absence of a normal cortisol awakening response (CAR) in adolescent males with Asperger syndrome (AS):
    https://www.ncbi.nlm.nih.gov/pubmed/19304400

    Consider this:

    With regards to low serotonin and loneliness, think of it this way, if you cannot feel loneliness (due to apathy/blunt affect) ofcourse there is a social wanting dysfunction, why would you want to be around others if you dont feel alone when socially isolated and have restricted interests.

    Basically we could describe aspergers as a 'lack of craving for oxytocin'.

    CDP-choline seems to benefit me by improving the response in the amygdala.

    ReplyDelete
  14. Sulforaphane seems to also targets the brain water regulating systems!!!!, it goes beyond nrf2/glutathione/antioxidant:

    Sulforaphane enhances aquaporin-4 expression and decreases cerebral edema following traumatic brain injury.
    https://www.ncbi.nlm.nih.gov/pubmed/16211562

    "Aquaporin-4 (AQP4) water channel is expressed at high levels in brain astrocytes, and the bidirectional transport of water through these channels is CRITICAL FOR THE MAINTENANCE OF BRAIN WATER HOMEOSTASIS."

    "Postinjury administration of sulforaphane (SUL), an isothiocyanate present in abundance in cruciferous vegetables such as broccoli, attenuated AQP4 loss in the injury core and further increased AQP4 levels in the penumbra region compared with injured animals receiving vehicle. These increases in AQP4 levels were accompanied by a significant reduction in brain edema (assessed by percentage water content) at 3 days postinjury. These findings suggest that the reduction of brain edema in response to SUL administration could be due, in part, to water clearance by AQP4 from the injured brain."

    ReplyDelete
  15. Oddly enough bumetanide seems to do the oposite??
    Can somewhat translate this to human words what this study says:

    Inhibition of aquaporin-1 and aquaporin-4 water permeability by a derivative of the loop diuretic bumetanide acting at an internal pore-occluding binding site.
    https://www.ncbi.nlm.nih.gov/pubmed/19403703

    ReplyDelete
  16. I just found another bumetanide study published last month:

    https://www.nature.com/articles/s41598-018-21958-x.pdf?origin=ppub

    Here we show that bumetanide treatment in autism normalizes amygdala activation in response to eye contact, and that it increases the time spontaneously spent in the eyes of dynamic emotional faces. These data complement our two previous studies that showed (1) increased activation in the social brain of participants with autism for emotional vs. neutral faces a er bumetanide treatment in an unconstrained paradigms, and (2) increase of subcortical face-processing network activation in autism compared to controls, including in the amygdala, when gaze is constrained in the eye region.

    1 mg per day for 10 months, but only 9 participants.

    ReplyDelete
    Replies
    1. Thanks for highlighting this study.

      The question I have is just what percentage of people with an autism diagnosis actually respond to bumetanide and are the apparent non-responders really gaining no benefit whatsoever, or in some cases the benefit is too subtle to get picked up in autism rating scales?

      Delete
  17. Wow krilloil.... Im blown away.

    So this has been day 2 on krilloil (only taking half the recommended dose) and let me tell you im absolutely blown away so far by this stuff, I was like 'meh' I'll give it a try as a replacement of regular fishoil. Let me you tell you (for me personally atleast) it is not on the same page as fishoil, hell its not even in the same book, the effect is very profound.

    Here are the ingredients, Im taking one capsule (which has half the content listed here).

    Im not sure if this due to astaxanthin, but my skin quality seems to dramatically improve aswell (regular fishoil never done this for me). The astaxanthin content in this seems low, but the phospholipids can greatly enhance the absorption.


    The supplement I got (was pretty cheap actually):
    https://ibb.co/dSB9Z7


    Astaxanthin alleviates cerebral edema by modulating NKCC1 and AQP4 expression after traumatic brain injury in mice

    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5007682/

    "Background
    Astaxanthin is a carotenoid pigment that possesses potent antioxidative, anti-inflammatory, antitumor, and immunomodulatory activities. Previous studies have demonstrated that astaxanthin displays potential neuroprotective properties for the treatment of central nervous system diseases, such as ischemic brain injury and subarachnoid hemorrhage. This study explored whether astaxanthin is neuroprotective and ameliorates neurological deficits following traumatic brain injury (TBI).

    Results
    Our results showed that, following CCI, treatment with astaxanthin compared to vehicle ameliorated neurologic dysfunctions after day 3 and alleviated cerebral edema and Evans blue extravasation at 24 h (p < 0.05). Astaxanthin treatment decreased AQP4 and NKCC1 mRNA levels in a dose-dependent manner at 24 h. AQP4 and NKCC1 protein expressions in the peri-contusional cortex were significantly reduced by astaxanthin at 24 h (p < 0.05). Furthermore, we also found that bumetanide (BU), an inhibitor of NKCC1, inhibited trauma-induced AQP4 upregulation (p < 0.05)."

    _____

    Effectiveness of astaxanthin in a valproic acid model of autism:

    Astaxanthin improves behavioral disorder and oxidative stress in prenatal valproic acid-induced mice model of autism.

    https://www.ncbi.nlm.nih.gov/pubmed/25732953

    _____

    @Peter
    Would this mean that astaxanthin works the same as bumetanide? Also they both affect aquaporin-4, sulforaphane also does this.

    ReplyDelete
    Replies
    1. Aspie, one of the many things that increases NKCC1 expression is inflammation. Astaxanthin acts by reducing the expression of NF-κB-mediated pro-inflammatory factors. If you are a bumetanide responder and have activated NF-κB, then astaxanthin should be helpful. But you might be a bumetanide responder and not have activated NF-κB.

      Bumetanide is an NKCC1 blocker while Astaxanthin reduces the number of NKCC1 transporters if NF-κB is activated, so the effect is similar but different.

      The question is how much Astaxanthin you would need, probably a lot.

      Delete
    2. Not sure, but from what ive noticed so far is that krilloil feels very very different than any fishoil supplement ive ever taken before.

      My skin quality is rapidly improving, literally in 2 days some yeast like symptoms seem to be cleaning up, this astaxanthin dose in it seems very very small though, but apparantly its a very potent antioxidant and is one of the few that can pass through the BBB.

      And nope I have not tried bumetanide yet (Im planning on though), quite a lot of pubmed results with it regarding to autism (especially the social problems which you know that is the one thing im actually trying to improve).

      Delete
    3. ..and as mentioned in an earlier discussion, Astaxanthin is a p38 inhibitor.
      I haven't been able to give it to my daughter (and am not very keen on it anymore due to the p38 inhibition which in our case seems to be the opposite to what we want). So instead I am popping a capsule a day for a couple of weeks back. I haven't noticed much, myself. Guess it has a very specific and beneficial effect for you Aspie!

      /Ling

      Delete
    4. Hi Ling,

      Im not sure if its down to the astaxanthin as there is only 100mcg! in 2 pills and I have been taking one pill so thats 50mcg astaxanthin/day. Now the phospholipids and fatty acids that are naturally present in krilloil do enhance absorption around 7-8 fold from what I have read in papers.

      Delete
    5. Aspie, maybe you should check up the Krill Oil + MFGM trail discussed in the comments here: https://epiphanyasd.blogspot.se/2018/03/autism-and-non-antibiotic-properties-of.html

      I have the Sports Research Krill oil that contains 1 mg of Astaxanthin per capsule, because this is the same manufacturer as in the study I try to copy.

      /Ling

      Delete
  18. Seems there is a possible overlap (atleast partially) between the efficiency of keto diet/bhb and bumetanide!
    It turns out bumetanide can activate GPR35, just like kyrunenic acid (often low in asd/autism and high in shizophrenia).

    https://en.wikipedia.org/wiki/Kynurenic_acid

    Kynurenine Pathway in Autism Spectrum Disorders in Children.
    https://www.ncbi.nlm.nih.gov/pubmed/29694960

    "RESULTS:
    The mean serum level of KA was significantly lower in the ASD group than in the healthy controls (28.97 vs. 34.44 nM, p = 0.040), while the KYN/KA ratio was significantly higher in the ASD group (61.12 vs. 50.39, p = 0.006). The same relative values were found when comparing the childhood autism subgroup with the controls. Also, the mean serum level of TRP was significantly lower in children with a subdiagnosis of childhood autism than in those with Asperger syndrome (67.26 vs. 77.79 μM, p = 0.020)"

    GPR35 Is a Target of the Loop Diuretic Drugs Bumetanide and Furosemide
    https://www.karger.com/Article/Abstract/335127

    Novel aspect of ketone action: β-hydroxybutyrate increases brain synthesis of kynurenic acid in vitro.
    https://www.ncbi.nlm.nih.gov/pubmed/20838951

    Ketogenic diet increases concentrations of kynurenic acid in discrete brain structures of young and adult rats
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3359463/

    Kynurenic acid is a nutritional cue that enables behavioral plasticity
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4334586/
    "Targeting mechanisms that result in increased concentrations of kynurenic acid (KYNA) in the brain has been considered as a therapeutic approach for the treatment of epilepsy and certain neurodegenerative disorders. Recently, KYNA has been implicated in the effects produced by the high-fat and low-protein/carbohydrate ketogenic diet (KD) in a report demonstrating an increased production of KYNA in vitro by one of the ketone bodies, β-hydroxybutyrate, elevated by the KD. To further explore this association, brain concentrations of KYNA were compared in young (3 weeks old) and adult (8–10 weeks old) rats that were chronically exposed to the KD and regular diet. Exposure to the KD resulted in the anticipated elevations of β-hydroxybutyrate with accompanying decreases in glucose concentrations. In comparison to rats fed the regular diet, KYNA concentrations were significantly (p < 0.05) increased in the hippocampus (256 and 363% increase in young and adult rats, respectively) and in the striatum (381 and 191% increase in young and adult rats, respectively) in KD-fed rats"

    Remember Peter saying to take panax ginseng to help with OAT function to potentiate bumetanide.
    Well panax ginseng (through a newly identified compound called gintonin) is a lysophosphatidic acid agonist and LPA ligands also lock onto GPR35

    Gintonin, a ginseng-derived lysophosphatidic acid receptor ligand, potentiates ATP-gated P2X1 receptor channel currents
    https://link.springer.com/article/10.1007/s10059-013-2293-x

    Gintonin, Newly Identified Compounds from Ginseng, Is Novel Lysophosphatidic Acids-Protein Complexes and Activates G Protein-Coupled Lysophosphatidic Acid Receptors with High Affinity
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3887723/

    GPR35 is a novel lysophosphatidic acid receptor.
    https://www.ncbi.nlm.nih.gov/pubmed/20361937

    The emerging pharmacology and function of GPR35 in the nervous system
    https://sci-hub.se/https://doi.org/10.1016/j.neuropharm.2015.07.035

    "Single nucleotide polymorphisms of GPR35 have linked this receptor to coronary artery calcification, inflammatory bowel disease and primary sclerosing cholangitis, while chromosomal aberrations of the 2q37.3 locus and altered copy number of GPR35 have been linked with autism, Albight’s hereditary osteodystrophy-like syndrome, and congenital malformations, respectively"

    Discovery of Natural Phenols as G Protein-Coupled Receptor-35 (GPR35) Agonists
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4025658/

    Some GPR35 ligands
    Baicalin = full agonist
    Quercetin and Luteolin = partial agonist

    ReplyDelete
  19. Happy new year to all!,

    Some interesting things regarding bumetanide and water regulation/behavior:

    Bumetanide blocks the acquisition of conditioned fear in adult rats. (2018)
    https://www.ncbi.nlm.nih.gov/pubmed/29235092

    "BACKGROUND AND PURPOSE:
    Bumetanide has anxiolytic effects in rat models of conditioned fear. As a loop diuretic, bumetanide blocks cation-chloride co-transport and this property may allow bumetanide to act as an anxiolytic by modulating GABAergic synaptic transmission in the CNS. Its potential for the treatment of anxiety disorders deserves further investigation. In this study, we evaluated the possible involvement of the basolateral nucleus of the amygdala in the anxiolytic effect of bumetanide.

    EXPERIMENTAL APPROACH:
    Brain slices were prepared from Wistar rats. extracellular recording, stereotaxic surgery, fear-potentiated startle response, locomotor activity monitoring and Western blotting were applied in this study.

    KEY RESULTS:
    Systemic administration of bumetanide (15.2 mg·kg-1 , i.v.), 30 min prior to fear conditioning, significantly inhibited the acquisition of the fear-potentiated startle response. Phosphorylation of ERK in the basolateral nucleus of amygdala was reduced after bumetanide administration. In addition, suprafusion of bumetanide (5 or 10 μM) attenuated long-term potentiation in the amygdala in a dose-dependent manner. Intra-amygdala infusion of bumetanide, 15 min prior to fear conditioning, also blocked the acquisition of the fear-potentiated startle response. Finally, the possible off-target effect of bumetanide on conditioned fear was excluded by side-by-side control experiments.

    CONCLUSIONS AND IMPLICATIONS:
    These results suggest the basolateral nucleus of amygdala plays a critical role in the anxiolytic effects of bumetanide."

    Bumepamine, a brain-permeant benzylamine derivative of bumetanide, does not inhibit NKCC1 but is more potent to enhance phenobarbital's anti-seizure efficacy. (december 2018)
    https://www.ncbi.nlm.nih.gov/pubmed/30248303

    "These data show that CNS effects of bumetanide previously thought to be mediated by NKCC1 inhibition can also be achieved by a close derivative that does not share this mechanism. Bumepamine has several advantages over bumetanide for CNS targeting, including lower diuretic potency, much higher brain permeability, and higher efficacy to potentiate the anti-seizure effect of phenobarbital."

    Neural correlates of sensory hypersensitivity in autism spectrum disorder
    https://www.sfari.org/funded-project/neural-correlates-of-sensory-hypersensitivity-in-autism-spectrum-disorder/

    parabrachial nucleus - https://en.wikipedia.org/wiki/Parabrachial_nuclei

    Why no talk about this before???

    2 Function
    2.1 Arousal (speaks for itself but also most likely can control seizures)
    2.2 Blood sugar control (autism and obesity connection)
    2.3 Thermoregulation (and thus PGE2)
    2.4 Taste (food aversion/sensitivities)
    2.5 Pleasure (reward, including social reward)

    ReplyDelete
  20. On top of that it seems to control water intake:


    A role for the lateral parabrachial nucleus in cardiovascular function and fluid homeostasis
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235290/

    "In addition, inactivation of the LPBN induces overdrinking of water in response to a range of dipsogenic treatments primarily, but not exclusively, those associated with endogenous centrally acting angiotensin II. Moreover, treatments that typically cause water intake stimulate salt intake under some circumstances particularly when serotonin receptors in the LPBN are blocked. This review explores the expanding body of evidence that underlies the complex neural network within the LPBN influencing salt appetite, thirst and the regulation of blood pressure."


    Activation of serotonergic 5-HT(1A) receptors in the lateral parabrachial nucleus increases NaCl intake.
    https://www.ncbi.nlm.nih.gov/pubmed/16360657

    "In contrast, WAY did not affect water intake produced by 8-OH-DPAT. WAY-100635 injected alone into the LPBN had no effect on NaCl ingestion. Injections of 8-OH-DAPT (0.1 microg/0.2 microl) into the LPBN also increased 0.3 M NaCl intake induced by 24-h sodium depletion (furosemide, 20 mg/kg bw plus 24 h of sodium-free diet). Serotonin (5-HT, 20 mug/0.2 mul) injected alone or combined with 8-OH-DPAT into the LPBN reduced 24-h sodium depletion-induced 0.3 M NaCl intake. Therefore, the activation of serotonergic 5-HT(1A) receptors in the LPBN increases stimulated hypertonic NaCl and water intake, while 5-HT injections into the LPBN reduce NaCl intake and prevent the effects of serotonergic 5-HT(1A) receptor activation."

    ReplyDelete
  21. Aspie1983, that Bumepamine article is very interesting. It's from the same group working on BUM5 that was mentioned a year ago or so here. What I find intriguing is that they claim to have found something that works better than bumetanide, but also tell us that they don't have a clue on why it works. They mention NKCC1 a couple of times, but not KCC2 which is odd - they must have read all the articles on Bumetanide. The big question is if they just are ignorant of the KCC2 effect, or if they actually think it is something else that does the trick?

    Thanks anyway for the link.

    /Ling

    ReplyDelete

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