Quantitative Testing of BAC's Effect on Bacterial Growth

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)
 
Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
 
RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
 
Anyone who skipped out on a college microbiology 101 course might still pick up a feel for how contamination risks work in a home or kitchen setting by spending some time on mycology as a hobby. Sure, contamination deserves attention, and there are straightforward, common sense steps to follow. But seriously, some of you carry on as though we were engineering gain of function pathogens in bats. What we're actually doing is reconstituting powders that should be properly lyophilized, using certified BAC, ideally with lab grade filters, and we have working immune systems. This exact process happens millions and millions of times every year — not only with peptides, but in clinics, on hospital wards, in the homes of diabetics, and so on. Truthfully, if you stick to normal precautions, the odds of running into a contamination problem are smaller than the odds of getting into a car crash on your commute.
 
bada bing said:

Anyone who skipped out on a college microbiology 101 course might still pick up a feel for how contamination risks work in a home or kitchen setting by spending some time on mycology as a hobby. Sure, contamination deserves attention, and there are straightforward, common sense steps to follow. But seriously, some of you carry on as though we were engineering gain of function pathogens in bats. What we're actually doing is reconstituting powders that should be properly lyophilized, using certified BAC, ideally with lab grade filters, and we have working immune systems. This exact process happens millions and millions of times every year — not only with peptides, but in clinics, on hospital wards, in the homes of diabetics, and so on. Truthfully, if you stick to normal precautions, the odds of running into a contamination problem are smaller than the odds of getting into a car crash on your commute.
Having laminar flow would also be a plus:

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To certain Europeans who skip BAC and don't filter, our concerns might come across as overblown, given that they simply rely on sterile water plus refrigeration.
 
Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
 
RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
 
Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
On top of that, if the gloves don't fit right, there's no point in bothering.
 
Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
In my opinion, the HEPA filter and the vent filter go beyond what's needed. For typical home use, that level of setup just isn't realistic. It's not that extra precautions cause harm, but the harder the routine is to follow, the less likely someone like Joe Blow at home will bother with it at all, and he might even point to the difficulty as a reason to skip everything. There's a narrow boundary between recognizing a risk and letting the process completely run your life.

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
Because each item comes in its own sterile packaging, I'm able to reconstitute and filter while never making contact with the parts of the instruments that need to stay clean. Yep, I'm cutting corners out here with no gloves on. 😆
 
RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
In my opinion, the HEPA filter and the vent filter go beyond what's needed. For typical home use, that level of setup just isn't realistic. It's not that extra precautions cause harm, but the harder the routine is to follow, the less likely someone like Joe Blow at home will bother with it at all, and he might even point to the difficulty as a reason to skip everything. There's a narrow boundary between recognizing a risk and letting the process completely run your life.

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
Because each item comes in its own sterile packaging, I'm able to reconstitute and filter while never making contact with the parts of the instruments that need to stay clean. Yep, I'm cutting corners out here with no gloves on. 😆
Yeah, that's overkill. During the 90's I worked as a microbiologist. A negative hood only came out when we were processing human stool or the odd other specimen. For work that truly required sterility, or when the material was especially hazardous, we turned to the Biosafety Cabinet/Box: you placed your items inside, then slid your arms into long gloves sealed at the glass, so whatever you handled within the box stayed essentially sterile.
 
Mr. B said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
In my opinion, the HEPA filter and the vent filter go beyond what's needed. For typical home use, that level of setup just isn't realistic. It's not that extra precautions cause harm, but the harder the routine is to follow, the less likely someone like Joe Blow at home will bother with it at all, and he might even point to the difficulty as a reason to skip everything. There's a narrow boundary between recognizing a risk and letting the process completely run your life.

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
Because each item comes in its own sterile packaging, I'm able to reconstitute and filter while never making contact with the parts of the instruments that need to stay clean. Yep, I'm cutting corners out here with no gloves on. 😆
Yeah, that's overkill. During the 90's I worked as a microbiologist. A negative hood only came out when we were processing human stool or the odd other specimen. For work that truly required sterility, or when the material was especially hazardous, we turned to the Biosafety Cabinet/Box: you placed your items inside, then slid your arms into long gloves sealed at the glass, so whatever you handled within the box stayed essentially sterile.
That said, your work wasn't in a kitchen lab, and you weren't a compounding pharmacist either.
 
Calm Logic said:

Mr. B said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
In my opinion, the HEPA filter and the vent filter go beyond what's needed. For typical home use, that level of setup just isn't realistic. It's not that extra precautions cause harm, but the harder the routine is to follow, the less likely someone like Joe Blow at home will bother with it at all, and he might even point to the difficulty as a reason to skip everything. There's a narrow boundary between recognizing a risk and letting the process completely run your life.

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

Calm Logic said:

RetCurious said:

If you filter right away once reconstitution is done, there wouldn't be much time for dead bacteria remnants to form, as far as I can tell. Is there something I'm overlooking here?
What classification of clean room is your setup using? That has consistently been the sticking point when it comes to filtering, and it still is today, even with group sterility testing now being a fairly routine alternative (notwithstanding variability within a batch).

(And even having a costly cleanroom did not prevent ProRx from being shut down by the FDA over careless operations.)

If the path runs from a filter, through a fresh sterile needle, and into a new sterile cartridge whose top has been swabbed properly, then the chance for contamination to sneak in seems quite small to me. Is there some angle here I'm not seeing?
Every surface is surrounded by air, and that air carries contaminants.
The move happens between two sealed setups, with the point of transfer cleaned and kept as brief as possible. In my view, whatever risk remains is minimal, perhaps even negligible.
What worries me most is the way even a tiny or negligible risk compounds as time goes on, particularly for those who are reconstituting several vials each week.

In the Peptide Test filtering video, gloves are worn, a HEPA filter is used, and a filter is placed on the vent needle. My own view is that gloves matter more than airflow does, though I could be wrong.
Because each item comes in its own sterile packaging, I'm able to reconstitute and filter while never making contact with the parts of the instruments that need to stay clean. Yep, I'm cutting corners out here with no gloves on. 😆
Yeah, that's overkill. During the 90's I worked as a microbiologist. A negative hood only came out when we were processing human stool or the odd other specimen. For work that truly required sterility, or when the material was especially hazardous, we turned to the Biosafety Cabinet/Box: you placed your items inside, then slid your arms into long gloves sealed at the glass, so whatever you handled within the box stayed essentially sterile.
That said, your work wasn't in a kitchen lab, and you weren't a compounding pharmacist either.
If you work as a compounding pharmacist, that makes sense. At home in the kitchen, though, you'll be fine as long as you take the proper sterile precautions.
 
Skidude said:

Here's what's been running through my mind regarding this:

When it comes to killing bacteria, BAC does the job well. However, the number and strain of organisms involved will determine how many Endos end up in the solution.

When only a tiny quantity of bacteria gets introduced, the fast kill rate appears to stop a large buildup of bacteria and Endos from taking hold.

On the other hand- should bacteria already be in there because the lyophilized powder wasn't sterile, that's where trouble could arise.

This appears to leave us with Static protection against what gets introduced, rather than any protection tied to sterility.

I'd really appreciate hearing what others think about this....
This is a misconception. The absence of visible bacterial cultures does not automatically prove the bacteria are dead. It may simply mean they are unable to reproduce, or at least not to a degree that becomes visible to the naked eye, during the period covered by the test.
 
Enyola said:

This is a misconception. The absence of visible bacterial cultures does not automatically prove the bacteria are dead. It may simply mean they are unable to reproduce, or at least not to a degree that becomes visible to the naked eye, during the period covered by the test.
I had the same understanding, and I'd come across the distinction between bacteriostatic and bactericidal.

That said, they were there, since they got added for the testing described here.

It goes against what you'd expect, yet the results of the tests say otherwise.

Could you lay out your reasoning for why bacteria being introduced and then no longer being detectable doesn't amount to killing?

I really am puzzled by this.

With no BAC present, they showed up right away.
 
These tests did not include any "immediately" condition.

Each sample was used to inoculate nutrient agar plates that were then incubated for 3 days to allow for good colony formation.

Click to expand...
Those 3 days had already passed before the photos were taken — nothing was captured right away. Individual bacteria are invisible to the naked eye; what shows up are the colonies formed once each one has multiplied through several generations.
 
Enyola said:

These tests did not include any "immediately" condition.


Those 3 days had already passed before the photos were taken — nothing was captured right away. Individual bacteria are invisible to the naked eye; what shows up are the colonies formed once each one has multiplied through several generations.
"immediately" when the measurement was taken-

Kindly keep the focus on the actual question.

I'm not trying to argue with you — I'm only after facts.

Take a look at the test results below-

At room temperature, after 1 DAY, nothing was detected in .9%.





05acfebccd42535818a1f684ec4aab5e5ef16413cb70dfe49c70123a33fd51b8.webp


this is how .3% appeared

40de8fae6d998aa0a9ef9e9a720120770ac5be15314596160e5e87d912e384f3.webp


How .9% appeared AFTER 1 DAY.

75f4fa9836742b47c5c1c1e0fbb6cbe0d6a2d8bd5212aad59640caacea7467f4.webp
 
My apologies — I had myself tangled up, and I'm withdrawing my earlier statement.

One thing matters a lot when looking at this test: the findings shouldn't be stretched beyond the scope of what was actually studied. Sensitivity to disinfecting chemicals varies enormously from one bacterial species to another. Skin bacteria were the ones chosen by the author. Those tend to fall among the more sensitive groups. At the opposite extreme, you have spores from genera such as bacillus or clostridium spp., which are considered resistant to benzyl alcohol. Somewhere in the middle sit other bacteria that might persist longer in a dormant state, until their systems shut down and they eventually starve.

Is there any information about which bacteria are present in typical grey vials? I haven't come across anything on that.
 
Enyola said:

My apologies — I had myself tangled up, and I'm withdrawing my earlier statement.

One thing matters a lot when looking at this test: the findings shouldn't be stretched beyond the scope of what was actually studied. Sensitivity to disinfecting chemicals varies enormously from one bacterial species to another. Skin bacteria were the ones chosen by the author. Those tend to fall among the more sensitive groups. At the opposite extreme, you have spores from genera such as bacillus or clostridium spp., which are considered resistant to benzyl alcohol. Somewhere in the middle sit other bacteria that might persist longer in a dormant state, until their systems shut down and they eventually starve.

Is there any information about which bacteria are present in typical grey vials? I haven't come across anything on that.
While the expectation is that filtering would help with whatever is present before reconstitution, it appears BAC additionally contributes to removing dangers rather than only stopping growth.
 
Skidude said:

While the expectation is that filtering would help with whatever is present before reconstitution, it appears BAC additionally contributes to removing dangers rather than only stopping growth.
Absolutely — the degree of bacterial 'elimination' is far greater than I had ever assumed. My understanding (shared by plenty of others, I suspect?) was that .9% BA merely held bacterial particles at a constant level...Reading this report has really opened my eyes, and it's useful information to have...

Even so, filtering into a sterile vial is something I'll keep doing every time. The idea of just leaving the solution sitting in its original vial — which I'm certain some people will soon be pushing for — is not something I can picture myself doing.... 😳 😎
 
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