156 replies to this topic

[resveratrol_guy]

 

This is Team TLR's response:

Please note we can afford the DNase1 material with strongest quality in the amount requested of 45 Mega kiu total material for $11,960.

 

Lead time is approximately 4-7 days.

this would be 20 grams at 1500ku. 

 

I'm sure the price is much higher than other options.  They have agreed to put it in escrow contingent on lab test results.  Finding the lap to conduct the tests has been the difficult.  I have very little background in this field and consider it a hobby.  I know much less about most of this stuff than nearly everyone on these boards.  With that said if someone could help me find someone to properly test this I would appreciate it.   Also if other people are interested at this price I'm happy to include them.  

 

 

According to the researchers, 1 mg contains 1500 KU. So 45 GU (45 MKU) should weigh 30 g. The patient consumed 120 mg/d, so this supply would last a single person 250 days @ $48/d.

 

And what does "strongest quality" mean?

Edited by resveratrol_guy, 03 July 2016 - 10:33 PM.

[resveratrol_guy]

 

I will re-Email them with a modified version of the above Email and see if we cant get a deep discount for our experiment RG.  

I doubt they are aware of the fact that the demand for Dnase may be about to skyrocket.

 

 

Psilociraptor1 has shed some light on how Dnase may work:

 

This is interesting though I'm having some internal conflict about it. Dr Judith Milkossy and Alan McDonald have also posted numerous studies identifying spirochetal infections in alzheimers lesions. My issue is that amyloids are also fundamental components of bacterial biofilms and therefor would be expected to bind to microorganisms without necessarily conferring immunity to the host. Unfortunately I do not have access to that full article, but I'm wondering if the authors are conflating "microbial entrapment" with intentional biofilm formation. There is also the possibility that certain bacteria exploit innate immune function and cross seed with endogenous amyloids making the distinction somewhat ambiguous. My personal feelings are that the presence of accumulated amyloids is more suggestive of biofilm formation than functional immune response. Interestingly I've noticed that many herbs that are reported to reverse beta-amyloid deposits are also potent biofilm erradicators. Ie flavonoids and related compounds such as baicelain and EGCG.

Interestingly the use of DNaseI is also a noted biofilm disrupter. I'm surprised to hear of in vivo efficacy though. Especially for a brain disorder.

 

Pretty much exactly what it sounds like. Something that disrupts biofilm development and or the structural integrity of mature biofilms. There is no "one" mechanism for it. DNases have been said to destabalize [biofilms] by cleaving some of DNA that's wound into the biofilm matrix. I don't know enough to say what their exact structural role is. Chelating agents can disrupt some biofilms that rely on iron for structure. Biofilm disruptors may also be proteolytic enzymes, quorum sensor inhibitors, agents which stimulate bacterial disaggregation pathways etc. This is a pretty understudied science and what works in vivo is almost completely unknown but these are the sorts of things studied in labs at the moment.

http://www.longecity...ndpost&p=780542

 

I haven't looked for any supporting studies of these statements..?

 

I think Biofilm disruptors and bacteria killers might synergise with Dnase..?

Some interesting anti pathogens:
http://www.longecity...ndpost&p=780706

 

 

I think Psilociraptor1 is probably on the right track. Presumably DNase 1 simply breaks down anything it finds containing DNA (except living cells, obviously, which are protected by the lipid bilayer). Depending on the particular cause of a given case of dementia, doing so might disaggregate enough plaque to make a major cognitive difference. As he suggested, it wouldn't need to break down the entire biofilm or plaque aggregate; it would just need to break it into small enough pieces to diffuse throughout the brain more evenly, so that neighboring neurons could once again communicate effectively. In a diseased brain with a compromised BBB, some of that waste could then drain into the circulatory system for excretion.

 

My main worry with all this is that we're mistaking an uncommon case of "bacterial Alzheimer's" for the common case of "plaque Alzheimer's". If that's true, then I would expect little to no benefit from this therapy. To be sure, there is only one way to find out.

[Mian Ali Ismail]

Will this also work on Viral or fungal causes as these two are also thought to cause neurodegeneration !

[Der Springende Punkt]

Today's pubmed alert contains a recent paper about Alzheimer's, spirochetes and biofilms (although with a very questionable claim "Alzheimer's disease (AD) is an infectious disease caused by spirochetes"):

 

Alzheimer's Disease: Assessing the Role of Spirochetes, Biofilms, the Immune System, and Amyloid-β with Regard to Potential Treatment and Prevention.

Edited by Der Springende Punkt, 04 July 2016 - 12:37 PM.

[resveratrol_guy]

Let's keep this moving. Can we settle on a supplier? It seems to me that TeamLTR is the most promising source at the moment. A reasonable fee ($300?) should be paid to the manager for the work, but we can debate about that. Logic, if you want to manage it, you have my vote.

 

@ Der Springende Punkt, this preprint article says the author presents a way to use penicillin to treat the disease, presumably by killing the spirochetes. Penicillin is virtually ineffective against anything these days, thanks to the emergence of resistant bacteria. So even if his theory is correct, the treatment seems unlikely to work in large populations. Having said that, it's clear that hyperactive immune response plays a role in AD, alongside copper and genetics. What we all want to know is whether clearing out the biofilms is enough to halt or reverse progression, even if the other issues are not addressed.

 

While this is certainly a test worth conducting, I'm not optimistic about the outcome, because presumably we would have noticed that cystic fibrosis patients taking Pulmozyme never get AD. The counter to that argument, though, is that they tend to be young and often do not live long enough (rarely over 60) to develop the disease. As I've said previously, it's likely that the clinical definition of AD will soon split into 2 or more branches, one for copper and heavy metals, and the other for infection.

 

For its part, Pulmozyme (dornase alfa) is used to break up mucus in the lungs which may be an immune response to infection. For example, about 70% of middle-aged CF patients are infected with Pseudomonas aeruginosa. But I find it hard to believe that this treatment would help in patients without infection, as mucus does not normally contain cfDNA. So it seems to me, given its empirical effectiveness, that it's likely breaking up biofilms in the lungs, which in turn signals to the immune system to back off because the pathogens seem to be decreasing in number, resulting in less mucus secretion and thus better oxygenation. Whereas in the brain, dissolving biofilms would presumably restore communication between neighboring neurons to some extent.

[Mian Ali Ismail]

Yes I agree lets keep Moving.This is a pretty good deal and we shall know the results in 2 months at most or even a month might be enough.There are many potential buyers interested.

 

They say Dnase1 cleaves the DNA and removes cell free DNA.

 

So how do we start ?

[Mian Ali Ismail]

Lets get this done

Edited by Mian Ali Ismail, 08 July 2016 - 03:24 AM.

[TRUGAN]

I might be interested. How is it stored or how long does it last before expired?

[deetown]

Why did you decide on that particular dose Mian Ali Ismail?

 

 I've been asking around about testing and not gotten very far.  I'm not even sure how to test a large enzyme like this.  Western blotting was mentioned to me by an associate.   I'm not familiar with the technique.  I've been contacting labs Today that I've found just from google searches and so far had no luck what so ever.  I would guess a lab could feed it some human DNA and see if it cleaves it properly.  I'd be willing to purchase a large amount up front for myself and tack on whatever anyone else wants if we could figure out testing.  

[Alex_G]

"This is the quotation of Dnase1 from Teamtlr.com. A months supply would cost close to $210 per person.

 

Yes we do provide DNase1 with highest quality.  It is 1600iu/mg and price depends on quantity.  

30mg at $199.95

100mg at $449.95

200mg at $799.95

500mg at $1399.95"

 

From the study the patient was given 40mg 3 times per day, so 120mg per day. that would be around $600 per day. How do you come up with $210 per month??

[Mian Ali Ismail]

@Mrwhite

 

It has to be shipped with dry ice and a specific temprature has to be maintained while shipping and storing.

 

@deetown

 

I think 120mg per person is ok as it had no sideeffects according to the researchers of the study.Regarding group buy I think if you buy a large amount you might not be able to sell most of it and it is best people buy of a group buy as it will save you a lot of hassel regarding shipping this product in dry ice 

[Mian Ali Ismail]

 

"This is the quotation of Dnase1 from Teamtlr.com. A months supply would cost close to $210 per person.

 

Yes we do provide DNase1 with highest quality.  It is 1600iu/mg and price depends on quantity.  

30mg at $199.95

100mg at $449.95

200mg at $799.95

500mg at $1399.95"

 

From the study the patient was given 40mg 3 times per day, so 120mg per day. that would be around $600 per day. How do you come up with $210 per month??

 

Sorry problem in my numbers !

Edited by Mian Ali Ismail, 08 July 2016 - 04:12 AM.

[Alex_G]

From your post in this group on June 14th:

In this case 40 mg of human recombinant DNase I (1500 KU/mg) was given orally three times a day in conjunction with the memantine therapy (10 mg daily). 

 

looks to be about the same KU as from the mentioned supplier.

I would be much happier with your numbers, but I don't get the math to work. :(

 

[Der Springende Punkt]

Be careful with TeamTLR. They have a bad reputation. They are said to only ship taurine labeled as every possible chemical.

Edited by Der Springende Punkt, 08 July 2016 - 08:40 AM.

[deetown]

Does anyone know the difference between Human Recominant Dnase I and other Recominant Dnase I?

[Alex_G]

I emailed one of the researchers about their use of DNase I. They have continued to keep their patient on the DNase I and so far so good, the patient has not reverted back.

I was hoping they had taken the patient off DNase and the patient hadn't reverted.

[Mian Ali Ismail]

@Deetown.

It was the Human Recombinant form of Dnase1 that was used in the study.

 

@Alex.

 

Could you ask them that can pulmozyme be given orally ?

[deetown]

TeamTLR is quoting the Bovine pancrease Dnase I.  The rhDnase seems to be from other sources.  If anyone knows the difference let me know.  

[Mian Ali Ismail]

Pulmozyme is minimally absorbed in rats.Does anybody here know about its oral Absorbtion in humans ?

[Logic]

I have 3 replies saying 'We'll look into it" the friendliest of which is:

...For the status of Pulmozyme, as I said previously, it will take 8 weeks for pilot experiment. I see you’re very eager, but it has only been one week. However, I will contact the lab, and then you, sometime mid-July to update each other on the status of the pilot.

As for legal status, Our legal department is looking into it. Once I hear back, I will definitely let you know as well.

I look forward to working with you further. Please feel free to continue corespondances with me, and ask any questions you have along the way. I will do my best to work with you fully.

 

It looks like this may take a while. I wish I knew who had it in stock!?
I hope someone finds somewhere.

The bovine form caused an allergic reaction of some sort in a study I scanned.

[TRUGAN]

I have 3 replies saying 'We'll look into it" the friendliest of which is:

...For the status of Pulmozyme, as I said previously, it will take 8 weeks for pilot experiment. I see you’re very eager, but it has only been one week. However, I will contact the lab, and then you, sometime mid-July to update each other on the status of the pilot.

As for legal status, Our legal department is looking into it. Once I hear back, I will definitely let you know as well.

I look forward to working with you further. Please feel free to continue corespondances with me, and ask any questions you have along the way. I will do my best to work with you fully.

 

It looks like this may take a while. I wish I knew who had it in stock!?
I hope someone finds somewhere.

The bovine form caused an allergic reaction of some sort in a study I scanned.

 

 

Im interested. How does group buy work? Does the supplier ship everyone's share directly to them or does one person have to receive it and then reship to the others?

[MarcB]

I would be interested in the DNase I group buy.

[deetown]

Latest from TeamTLR.  I can at least say that it's a good sign they aren't trying to sell me fake product.  Unfortunately this doesn't bode well for our chances of getting rhDnase at any point. 

 

 

I have reached out to all companies and associates with potential for garnering best supply and price for rhDNase1. 

 

Indeed, as so noted prior, it is quite expensive to produce; as it is even at the very best price far more than the bovine form (bpDNase1).  The best price I could get affords the offer as below.

 

Quote is as follows:

 

30g $56,000USD

60g $98,000USD

 

Notably, all other pricings received were far higher. Pulmozyme likely can only be produced at what is potentially a somewhat lower cost basis due to mainly the huge scale and  as well production refinements. Both are such that only a big pharma company can potentially achieve with a rh protein such as this, though they even may have a similar cost basis as to the basis I was afforded and relates to the offer.   Obviously though, the offers as above given are still over 80% lower than the cost basis in comparison to retail price of Pulmozyme.

 

I wish it were possible to be produced at a better rate, but regrettably this is appearing to be by far the best offer.  Kindly let me know how you wish to proceed.

 

Thank you~

 

 

 

 

[Mian Ali Ismail]

I got this Email from the researcher who conducted the trial.So plain oral administration which is available in liquid form  wouldnt be helpful and we cant put the liquid in capsule form.WHat should I ask the researcher for any questions regarding administration in IV/IM form ?

 

Dnase (Pulmozyme) is an recombinant protein and as you know proteins may be destructed in the stomach and intestine. That is why many proteins are given IV/IM or per os in the form of capsules.  I hope it would be helpful.

[Major Legend]

I would be interested in this if it wasn't impossible to convince my grandparents that I am not trying some crazy.

 

Transport sounds like it will be an issue, biologics are not very stable substances. The capsules may have to be specifically designed, or at the very least have some kind of delivery technology e.g hydrogel.

[Mian Ali Ismail]

I think IV/IM route is much better .But what should I ask them about the protocol to follow for IV/IM route ?

[resveratrol_guy]

Just a thought... since IV/IM is a whole problem in and of itself, could we just take a proton pump inhibitor to shut down acid production, then take it orally before production resumes? These drugs are available over the counter or can easily be obtained through a doctor in most countries.

 

My other thought, given the enormous expense in question, is that we should just look for cheap Pulmozyme that we can still trust. For example, many drug companies sell their drugs cheaper in third world countries.

 

Maybe asking the labs is the wrong approach because we need bacteria (and hamsters, apparently) to synth it. It's way more complex and subject to error than producing a small molecule such as resveratrol.

 

Have we tried hard enough to find cost-effective Pulmozyme? Seriously, for the money we're talking about here, it would be more practical to fly half way around the world to buy it, then stay in a hotel for a week or two while dosing. (Pulmozyme party, anyone? Ha!) In theory, it should cause sustained cognitive remediation because unless there's active infection in the brain, we wouldn't expect the cfDNA to magically return after dosing cessation.

 

[Mian Ali Ismail]

This thread seems to be moving too slow.I heard that Dnase1 was also given IV for kidney disease in Israel.can anyone find the paper published or an article about it ?

[Linda Gray]

http://www.ncbi.nlm....pubmed/27073834
Is this what you are looking for?

[deetown]

Another update from TLR.  

 

Hi Mischa,

After receiving what was by far the best quote for rhDNase1 and remitting the offer to you, bearing I knew it was still likely higher than what was feasible, I subsequently inquired to a colleague of mine who specializes in protein expression optimization as to his thought to creating an optimized system and within that what best cost basis could be garnered. Usually he gets around $4000 upfront for this service, but as we both do many favors for each other and that he strongly supports Project TLR he was willing to explore it at no charge. 

Within this, his initial assessment is that an optimized expression system that he can derive should be able to provide a cost basis that is about half of the prior pricing offered. 

 

If something of that nature is workable for you I will inform him to proceed. At this point he does not know the actual firm cost basis with certainty, but he seems confident within pursuing it he can get the expression to yield means for production that produces the ability to offer a price that is around that price basis.

 

So, let me know if this seems to be within the area that will be viable.  Such would apparently seem to have ability to be under $1/mg basis, which I hope is tenable.  If so, he will than work to refine the expression and look to achieve a best cost basis that should hopefully be somewhere in accord with this initial estimated cost basis.  Once such is established, from there I will remit the firm offer to you based on such. 

Brgds~