Enteroviruses - revisited

[sometexan84]

Presumably by sending the serum sample directly to ARUP.

You do not have to go through LabCorp or Quest to arrange ARUP test, you can send the sample directly to ARUP, and pay for it yourself.

If you are outside the US, that's the way you would do it. I believe it is only if you have US health insurance that you have to go via LabCorp or Quest.

I'm in the US. I'm paying out of pocket anyway. Is there a place on their website that talks about sending samples directly, and paying ARUP?

 

[Hip]

Is there a place on their website that talks about sending samples directly, and paying ARUP?

Not that I am aware of.

But some years ago I emailed ARUP Lab, saying that I wanted to submit a serum sample for testing, and they sent me back all the info I needed: their address to send specimens, shipping instructions, forms to fill out, and prices.

So I think if you email them, they should send you this info.

 

[sometexan84]

Well, that didn't work...

 

[Hip]

Well, that didn't work...

View attachment 40415

Oh well. Possibly ARUP have changed their policy. Maybe though you might find a lab which has an account with ARUP that might be more competent in sending blood samples than LabCorp or Quest.

This is the email ARUP sent me in 2014 when I enquired about sending samples to them from the UK:

Spoiler: Email from ARUP

Thank you for inquiring about testing at ARUP Laboratories. Included in this e-mail you will find Test/specimen requirements and information, a blank ARUP requisition, shipping instructions and options for payment. We appreciate your business and hope you continue to use ARUP Laboratories as your reference laboratory of choice.

Thank you for prepaying these tests

Test/Specimen Requirements and information (Price is only valid for this submission)

Coxsackie B Virus Antibodies

Price: US$ 438.00

Orderable Test Num: 0060055
Methodology: Semi-Quantitative Serum Neutralization
Reported: 6-9 days
Collection: Serum separator tube or plain red. OR CSF.
Specimen Preparation: Separate serum from cells within 2 hours of collection. Transfer 1 mL serum or CSF to an ARUP Standard Transport Tube. (Min: 0.3 mL) Parallel testing is preferred and convalescent specimens must be received within 30 days from receipt of acute specimens. Mark specimens plainly as "acute" or "convalescent."
Storage/Transport Temperature: Refrigerated.
Unacceptable Conditions: Contaminated, hemolyzed, or severely lipemic specimens.
Stability (collection to initiation of testing): Ambient: 48 hours; Refrigerated: 2 weeks; Frozen: 1 year (avoid repeated freeze/thaw cycles)



Echovirus Antibodies

Price: US$ 445.00

Orderable Test Num: 0060053
Methodology: Semi-Quantitative Serum Neutralization
Reported: 6-9 days
Collection: Serum separator tube or plain red. OR CSF.
Specimen Preparation: Separate serum from cells within 2 hours of collection. Transport 1 mL serum or CSF. (Min: 0.3 mL) Parallel testing is preferred and convalescent samples must be received within 30 days from receipt of acute samples. Mark samples plainly as "acute" or "convalescent."
Storage/Transport Temperature: Refrigerated.
Unacceptable Conditions: Plasma.
Stability (collection to initiation of testing): Ambient: 48 hours; Refrigerated: 2 weeks; Frozen: 1 year (avoid repeated freeze/thaw cycles)


ARUP Requisition

The ARUP Requisition must be completed and submitted to ARUP laboratories along with the specimen. The ARUP Requisition must be filled out completely with the patient name and date of birth on the requisition, matching exactly with the specimen. If the requisition is not submitted to ARUP Laboratories with the specimen or if the patient name / Date of birth on the requisition do not match the specimen, there could be a delay in testing.

Shipping Instructions

Specimen and ARUP requisition should be shipped at sender’s expense to:

Genetics Processing 393 /
500 Chipeta Way, SLC, UT, 84108

Please inquire with local customs into shipping guidelines

Pre-Payment Options

Payment must be submitted prior to specimen/order receipt at ARUP Laboratories. If payment has not been received prior to specimen arrival, testing and results will be delayed until payment can be reconciled.


Purchase order number, Billing Contact Name, and Billing Address:

Credit Card (Please e-mail receipt from PayTrace application to Internationalsubmissions@aruplab.com

https://paytrace.com/cart/donate.pay?MID=555546008700&ID=10991&PID=Invoice


Bank Wire
Zions Bank, Foothill Branch
PO Box 27964
Salt Lake City, Utah 84127
Routing number: 124000054
Account Number: 91401513
Account Name: ARUP Laboratories
Checking Account
SWIFT Routing Number ZFNBUS55

Email for remittances: Internationalsubmissions@aruplab.com

Thanks,

Sergio M. Morales
Problem Resolution Specialist
Integrated Genetics/Oncology Services

500 Chipeta Way, Salt Lake City, Utah 84108-1221Phone: (801) 583-2787 ext.3303
Fax: (801) 584-5249
E-mail: sergio.morales@aruplab.com
Web: <www.aruplab.com>

 

[mrmichaelfreedmen]

@pete_parker I just found two other online Russian pharmacies which sell interferon alpha products, and interferon gamma products.

In Dr Chia's interferon therapy tests, ME/CFS patients given the interferon alpha plus gamma combo seemed to last the longest before relapse (14 months in some cases).

Pavel (OTC) from your above link is excellent

 

[Hip]

Pavel (OTC) from your above link is excellent

Yes, https://otc-online-store.com is a good pharmacy for Russian and other drugs. Been in business since 2013.

 

[Kalina]

Are there any news on the antiviral meds against coxsakie B? Will they come out this year or did the coronavirus stop their development?

 

[Hip]

Are there any news on the antiviral meds against coxsakie B? Will they come out this year or did the coronavirus stop their development?

I've heard no news at all. Usually the news I hear comes from patients of Dr Chia, because Chia is in the loop regarding the drug development (it was actually Dr Chia who is responsible for instigating the marketing of these Rega Institute antiviral drugs, as he explained to the pharmaceutical companies that they could be used to treat ME/CFS, which made them interested).

 

[Cipher]

@Hip

N-Acetyl cysteine effectively alleviates Coxsackievirus B-Induced myocarditis through suppressing viral replication and inflammatory response

Abstract

Viral myocarditis caused by Coxsackievirus B (CVB) infection is a severe inflammatory disease of the myocardium, which may develop to cardiomyopathy and heart failure. No effective specific treatment is available. Our previous study demonstrated that suppression of proinflammatory caspase-1 activation effectively inhibited CVB replication. N-acetyl cysteine (NAC) is a widely used antioxidant. In this study, we found that NAC significantly alleviated the myocardial injury caused by CVB type 3 (CVB3) under in vivo condition. Importantly, NAC treatment simultaneously suppressed viral replication and inflammatory response in both myocardium and cell culture. The antiviral and anti-inflammation mechanism of NAC, while independent of its antioxidant property, relies on its inhibition on caspase-1 activation. Moreover, NAC promotes procaspase-1 degradation via ubiquitin proteasome system, which further contributes to caspase-1 down-regulation. NAC also inhibits the activity of viral proteases. Taken together, this study shows that NAC exerts potent anti-CVB and anti-inflammation effect through targeting caspase-1. Given that NAC is a clinically approved medicine, we recommend NAC as a valuable therapeutic agent for viral myocarditis caused by CVB.

15 mg/kg (body weight) of NAC was given intraperitoneally to mice twice a day beginning at 12 h of p.i. Mice were euthanized at the end of day 5 of p.i.

That should be equivalent to around 1700-2847 mg NAC per day for someone who weighs 70kg:

Spoiler

15mg/kg * 2 (twice daily) = 30mg/kg
30/12,3 (mice -> human conversion) = 2,44 mg/kg
2,44 * 70kg = 170,8 mg
170,8 / 0,1 (6-10% oral bioavailability) = 1708mg/day
170,8 / 0,06 (6-10% oral bioavailability) = 2847mg/day

Could it work for non-cytolytic CVB infections?

 

[Hip]

Could it work for non-cytolytic CVB infections?

Very interesting study, I had not seen this one before.


My guess is that N-acetyl-cysteine (NAC) could work for non-cytolytic coxsackievirus B infections, because the study showed that NAC was able to inhibit Coxsackie B virus RNA replication:

Viral RNA (Fig. 2D), as well as the production of virions (Fig. 2E) were significantly reduced in the cells treated with NAC. These results indicate that NAC exerts potent inhibition on CVB3 replication.

Antivirals which inhibit virion entry into cells I believe will not work for non-cytolytic infection, because with non-cytolytic infection, the virus is already living inside the cell as naked RNA, and this RNA self-replicates in the cell.

But since NAC inhibits the replication of the viral RNA inside the cell, in theory I believe this could be useful for non-cytolytic coxsackievirus B.


However, the question is, what dose of NAC would be effective against this low-level of coxsackievirus B RNA found in the cells of ME/CFS patients?

By your above mouse-to-human calculation, a human oral dose of around 1 gram of NAC twice daily would be effective for acute coxsackievirus B infection; but I wonder if that dose would be sufficient for non-cytolytic infection, which replicates much more slowly?

I have taken NAC 600 mg twice daily for periods of weeks and months, mainly for its antioxidant effects (NAC increases levels of intracellular glutathione). But at that dose level, I never noticed any improvements in my ME/CFS.

Possibly trying around 10 times the dose might result in benefits: say 5 grams twice daily. You can buy NAC as a bulk powder.

Here it says:

commonly used maximum doses for IV acetylcysteine are 150 mg/kg (Max: 15,000 mg)

But this maximum dose is for short term use.


The study also tested NAC in some in vitro experiments in HeLa cells, and determined the maximum viral RNA inhibition was achieved at a concentration of 20 mM = 20,000 μM. That corresponds to a concentration of 3260 μg/ml = 3.2 mg/ml (since μg/ml = μM x molecular weight / 1000).

That is a very high concentration to achieve in the blood and body fluids. If we assume 40 liters = 40,000 ml of fluids in a human body, it would require an injectable dose of 128 grams to reach this 3.2 mg/ml concentration in the blood (and that does not even take into account NAC's plasma protein binding of 66-97%, which would necessitate an even higher dose).

 

[godlovesatrier]

Why do you think 5 grams @Hip ? I've never dabbled with nac. Just currently getting into it now. Titrating up currently as nac knocks me out in high doses

 

[Hip]

Why do you think 5 grams @Hip ? I've never dabbled with nac. Just currently getting into it now. Titrating up currently as nac knocks me out in high doses

Well just because I never noticed any ME/CFS benefits from taking NAC at 600 mg twice daily, and I have active coxsackievirus B4 infection. Many ME/CFS patients have tried NAC at around this dose level. So this dose level would appear to be too low for any potential antiviral effects.

So possibly moving up to a higher dose level of around 10 times greater, eg, around 5 grams twice daily, might start showing benefits.

I am not sure if there might be any adverse effects taking a dose that high for a prolonged period (I would guess you may need to take it for a month or two to notice any antiviral effects).

 

[godlovesatrier]

I get you. Could be true. With myself I can usually rely on feeling a bit pants if it has an anti bac or anti viral effect. Usually once that clears I feel a lot better. But beyond that it's hard to guage the benefits.

Even so NAC at higher doses might confer some more energy benefits and/or neuro benefits? Which can only be good.

I'm just thinking that if high dose thiamine doesn't really kick in until 1200mg (for me anyway) and the researchers and Corts surveys showed 1500 to 1800mg. Then there might be an ideal therapeutic dose for NAC. Joshua mentions cysteine in his mitochondrial disease papers for herpes veridea in fairly high doses 3x a day. He's had quite a few successes so far in private patient trials.

In v3.1 of his protocol not to be confused with the mitochondrial paper for herpes veridea he suggests 1000mg 3x a day. If you message him he might be able to explain the scientific reason behind this dosage? He's open to questions.

 

[Cipher]

So possibly moving up to a higher dose level of around 10 times greater, eg, around 5 grams twice daily, might start showing benefits.

I am not sure if there might be any adverse effects taking a dose that high for a prolonged period (I would guess you may need to take it for a month or two to notice any antiviral effects).

It seems like higher doses of NAC are safe (apart from potential nausea) and have more potent effects compared to lower doses according to these studies:

N‐acetylcysteine reduces disease activity by blocking mammalian target of rapamycin in T cells from systemic lupus erythematosus patients: A randomized, double‐blind, placebo‐controlled trial (2012)

Abstract

Objective
Systemic lupus erythematosus (SLE) patients exhibit T cell dysfunction, which can be regulated through mitochondrial transmembrane potential (Δψm) and mammalian target of rapamycin (mTOR) by glutathione (GSH). This randomized, double‐blind, placebo‐controlled study was undertaken to examine the safety, tolerance, and efficacy of the GSH precursor N‐acetylcysteine (NAC).
Methods
A total of 36 SLE patients received either daily placebo or 1.2 gm, 2.4 gm, or 4.8 gm of NAC. Disease activity was evaluated monthly by the British Isles Lupus Assessment Group (BILAG) index, the SLE Disease Activity Index (SLEDAI), and the Fatigue Assessment Scale (FAS) before, during, and after a 3‐month treatment period. Mitochondrial transmembrane potential and mTOR were assessed by flow cytometry. Forty‐two healthy subjects matched to patients for age, sex, and ethnicity were studied as controls.
Results
NAC up to 2.4 gm/day was tolerated by all patients, while 33% of those receiving 4.8 gm/day had reversible nausea. Placebo or NAC 1.2 gm/day did not influence disease activity. Considered together, 2.4 gm and 4.8 gm NAC reduced the SLEDAI score after 1 month (P = 0.0007), 2 months (P = 0.0009), 3 months (P = 0.0030), and 4 months (P = 0.0046); the BILAG score after 1 month (P = 0.029) and 3 months (P = 0.009); and the FAS score after 2 months (P = 0.0006) and 3 months (P = 0.005). NAC increased Δψm (P = 0.0001) in all T cells, profoundly reduced mTOR activity (P = 0.0009), enhanced apoptosis (P = 0.0004), reversed expansion of CD4−CD8− T cells (mean ± SEM 1.35 ± 0.12‐fold change; P = 0.008), stimulated FoxP3 expression in CD4+CD25+ T cells (P = 0.045), and reduced anti‐DNA production (P = 0.049).
Conclusion
This pilot study suggests that NAC safely improves lupus disease activity by blocking mTOR in T lymphocytes.

Glycine and N‐acetylcysteine (GlyNAC) supplementation in older adults improves glutathione deficiency, oxidative stress, mitochondrial dysfunction, inflammation, insulin resistance, endothelial dysfunction, genotoxicity, muscle strength, and cognition: Results of a pilot clinical trial (2021)

Abstract
Background
Oxidative stress (OxS) and mitochondrial dysfunction are implicated as causative factors for aging. Older adults (OAs) have an increased prevalence of elevated OxS, impaired mitochondrial fuel‐oxidation (MFO), elevated inflammation, endothelial dysfunction, insulin resistance, cognitive decline, muscle weakness, and sarcopenia, but contributing mechanisms are unknown, and interventions are limited/lacking. We previously reported that inducing deficiency of the antioxidant tripeptide glutathione (GSH) in young mice results in mitochondrial dysfunction, and that supplementing GlyNAC (combination of glycine and N‐acetylcysteine [NAC]) in aged mice improves naturally‐occurring GSH deficiency, mitochondrial impairment, OxS, and insulin resistance. This pilot trial in OA was conducted to test the effect of GlyNAC supplementation and withdrawal on intracellular GSH concentrations, OxS, MFO, inflammation, endothelial function, genotoxicity, muscle and glucose metabolism, body composition, strength, and cognition.
Methods
A 36‐week open‐label clinical trial was conducted in eight OAs and eight young adults (YAs). After all the participants underwent an initial (pre‐supplementation) study, the YAs were released from the study. OAs were studied again after GlyNAC supplementation for 24 weeks, and GlyNAC withdrawal for 12 weeks. Measurements included red‐blood cell (RBC) GSH, MFO; plasma biomarkers of OxS, inflammation, endothelial function, glucose, and insulin; gait‐speed, grip‐strength, 6‐min walk test; cognitive tests; genomic‐damage; glucose‐production and muscle‐protein breakdown rates; and body‐composition.
Results
GlyNAC supplementation for 24 weeks in OA corrected RBC‐GSH deficiency, OxS, and mitochondrial dysfunction; and improved inflammation, endothelial dysfunction, insulin‐resistance, genomic‐damage, cognition, strength, gait‐speed, and exercise capacity; and lowered body‐fat and waist‐circumference. However, benefits declined after stopping GlyNAC supplementation for 12 weeks.
Conclusions
GlyNAC supplementation for 24‐weeks in OA was well tolerated and lowered OxS, corrected intracellular GSH deficiency and mitochondrial dysfunction, decreased inflammation, insulin‐resistance and endothelial dysfunction, and genomic‐damage, and improved strength, gait‐speed, cognition, and body composition. Supplementing GlyNAC in aging humans could be a simple and viable method to promote health and warrants additional investigation.

OAs were provided capsules of glycine (1.33 mmol/kg/day) and cysteine (0.81 mmol/kg/day, provided as N‐acetylcysteine [NAC]) prepared by a licensed pharmacist ...

The glycine dosage is equivalent to 0.1 mg/kg/day (7g/day for someone who weighs 70kg). When it comes to the NAC dosage I'm a bit unsure what they mean, 0.81mmol of cysteine or N‐acetylcysteine. It's either 0,098mg/kg/day (6,86 g for 70 kg weight) or 0,132 mg/kg/day (9,24 g for 70 kg weight). From what I can see they didn't specify how many times a day they took the NAC/glycine.

To minimize nausea it might be prudent to not take more than 1.2 grams of NAC at a time, as 2.4 grams per dose gave 33 % of patients nausea in the first study.

 

[Pyrrhus]

Possibly relevant:

Enhanced glutathione levels may protect against some infections:
Role of glutathione in immunity and inflammation in the lung (Ghezzi, 2011)
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3048347/

Supraphysiological doses of cysteine (NAC) have a sedative effect:
(currently a popular use of high-dose NAC in psychiatry)
Cystine/Glutamate Exchange Regulates Metabotropic Glutamate Receptor Presynaptic Inhibition of Excitatory Transmission and Vulnerability to Cocaine Seeking (Moran et al., 2005)
https://www.jneurosci.org/content/25/27/6389.long

 

[sometexan84]

@Hip

Regarding the NAC... what if it was given IV?

 

[Hip]

Regarding the NAC... what if it was giving IV?

That's an idea to explore, though given NAC bulk powder is cheap, and its oral bioavailability is approximately 10%, it may just be easier to give an oral dose 10 times the intended intravenous dose.

I am not sure of the maximum dose of NAC that is safe for continuous long term use. For paracetamol poisoning, 10 grams of intravenous NAC is given, but this is a one-off dose.

I found this study of a case of fatal poisoning from an intravenous NAC overdose: a nurse in hospital had accidentally administered 100 grams of intravenous NAC rather than the 10 grams she was supposed to give, and the patient died of the adverse effects 12 days later.

 

[Hip]

The authors of the NAC Coxsackie B virus study seem to think N-acetyl-cysteine's inhibition of caspase-1 activation is the main antiviral mechanism of NAC for CVB. They say:

we believe that the activation of caspase-1 is crucial for CVB3 replication

So taking other caspase-1 inhibitors with NAC may boost its antiviral effects. This study says that some flavonoid supplements inhibit caspase-1, see table 2 of the paper.

Myricetin looks the most potent in that table, as it as the lowest Ki value (strongest effect). Whether it is possible from oral dosing to achieve blood concentrations that are able to inhibit caspase-1, that is a more difficult question.

There is also a caspase-1 inhibitor drug called belnacasan (VX-765) available, which Wikipedia says has proved well tolerated in human clinical trials.

 

[sometexan84]

I found this study of a case of fatal poisoning from an intravenous NAC overdose: a nurse in hospital had accidentally administered 100 grams of intravenous NAC rather than the 10 grams she was supposed to give, and the patient died of the adverse effects 12 days later.

Oh my god.

 

[sometexan84]

Regarding Enterovirus (testing, results, labs), I've created a poll. I'm hoping to get enough responses so we can see how common it is, and maybe learn more about testing accuracy.

If you get a moment, please have a look and respond - https://forums.phoenixrising.me/thr...r-enterovirus-b-which-lab-results-poll.83744/