Era/is
Rising Star
A problem that may not concern those who use sporadically and limited to one day of inhibitors, but which could perhaps involve those who use the various forms of inhibitors: natural or pharmaceutical.
Being a period of plant harvesting, I dedicated myself to the browning and after a period of intake, I found headaches that arises about two hours after they lay down.
Having already experienced some illnesses originating from the Mao inhibitors, I can report that those of the browning would seem selective. They probably interact with mao-b enzymes.
The problem is that by reading on the web what substances act as imao, you wonder why they are products far from feeding. I explain better: our body contains enzymes that oxidize amines and later with age, our body would produce more - perhaps because the amines deflect the mind a bit.
I wonder if drugs acting as inhibitors lead to well-being or not. The periods that you find yourself with inactive enzymes are marked by a general malaise that does not give a glimpse of their pharmaceutical use: headaches, exhaustion, general malaise are the order of the day.
Probably someone will be able to describe more completely their physical and soul state, if they take some psychopharmaceuticals, drugs or constantly use imao on an experimental basis.
From my point of view, you can perhaps notice the increase in vitality for an early period. Once the amines can act without being oxidized, they perform on the body and the psyche as a feeling of stillness that turns into anger if stressed; brief moments of euphoria; non-elastic body - muscle stiffness; disinterest and more.
On some sites I have found various discrepancies regarding the functions of mao enzymes, which leads to the question of whether using substances that make them ineffective is correct. By that I mean to question the constant use, not the sporadic one (from what I have been able to ascertain, to return to the 'normal' situation, it takes months and not weeks).
For example, with well-active enzymes, tyrmine acts as an invigorating one. Serotonin, however, works by giving happiness, if of genuine origin - that is, derived from precursors. The dmt is still active, even if it is a matter of wanting.
Is there anyone who wants to 'disassemble' the aspects of something still little known?
MAOi lead to an indirect increase in amines in the brain whether they are endogenous or external.
Our body tends to rebalance an excess of these amines, ES. serotonin, so when the substance finishes its effect often the balance of these amines is negative and so you have all the various symptoms like headaches, etc...
MAOi are very safe, whether they are of natural origin or of synthesis, problems arise when they are paired with releaser substances such as amphetamines always with regard to serotonin.
In this case the "extra" neurotransmitters cannot be removed from the body because the enzymes are blocked and therefore the effect is extremely more powerful and more durable, but also the risk of serotonin syndrome increases considerably.
Different is the case of DMT and all psychedelic triptammines in fact these molecules are amines and if the enzymes that should oxidize and destroy them are inhibited not only neurotransmitters can not be degraded but the molecule itself remains active for longer.
Extraction of MAO:
https://www.sciencedirect.com/science/a ... 2810000069
,then:
MONOAMINE OXIDASE EXTRACTION
Thaw the samples and centrifuge at 5000 rpm for 10 minutes at 4 ºC.
Wash the mitochondria with “cloreto de potássio 1,15%”.
Centrifuge at 5000 rpm for 10 minutes at 4 ºC.
Wash the purified mitochondria with “tampão de fosfato de potássio 0,1 M, pH 8,2, com Triton
X-100 0,5%”.
Centrifuge during 20 minutes at 10 000 rpm (4 ºC).
Suspend and homogenize the washed mitochondria with “tampão de fosfato de potássio 0,1 M,
pH 8,2, com Triton X-100 1%, colato de sódio 0,5% e -mercaptoetanol 0,002 M” (use an ice
bucket).
Assay for monoamine oxidase and compare with the spectrum from Figure 1.
Maintain the extract with gentle stirring in the cold for 4 hours.
Freeze the samples at – 20 ºC.
ION EXCHANGE CHROMATOGRAPHY
• Packing the column:
Put in the bottom of the column a piece of cotton and a circular net to retain the gel.
Dilute 30 mL of DEAE-Sephacel with 10 mL of equilibrium buffer (“tampão de fosfato de
potássio 0,01 M, pH 7,4, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M”). Put the
slurry of gel in the column slowly.
Start the flow; 1 to 2 mL/min is sufficient (1 drop every 2 seconds)
Note: the slurry of gel cannot be exposed to air!
The column should be equilibrated with 50 mL of equilibrium buffer (“tampão de fosfato de
potássio 0,01 M, pH 7,4, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M”).
• Enzyme dilution:
Dilute the enzyme to about one-and-a-half volume with “tampão de fosfato de potássio 0,01 M,
pH 7,6, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M” (25 mL of enzyme to
37,5 mL of buffer).
• Chromatography:
Elute 10 mL of “tampão de fosfato de potássio 0,02 M, pH 7,6, contendo Triton X-100 a 0,1% e
-mercaptoetanol a 0,002 M”.
Elute 4 mL of the diluted enzyme by employing a sodium chloride gradient (0.0 to 0.5 M) in
“tampão de fosfato de potássio 0,02 M, pH 7,6, contendo Triton X-100 a 0,1% e -
mercaptoetanol a 0,002 M”.
Collect manually 4 mL of eluted enzyme in separate tubes at a flow rate of 1 mL/min.
Read the content of the different tubes at 280 nm. Gather the fractions with higher absorbance.
Wash the column with 10 mL of “tampão de fosfato de potássio 0,02 M, pH 7,6, contendo Triton
X-100 a 0,1% e -mercaptoetanol a 0,002 M”.
Repeat the procedure until all the volume of the dilute enzyme is eluted.
• Ammonium sulfate fractionation
Measure the total volume of the combined fractions and determine the amount of ammonium
sulfate needed for 45% ammonium sulfate saturation (Table I).
Centrifuge at 10 000 rpm, during 25 minutes at 4 ºC.
Dissolve the enzyme in 3 mL of “tampão de fosfato de potássio 0,02 M, pH 7,4, contendo Triton
X-100 a 0,1% e -mercaptoetanol a 0,002 M”.
• Dialysis
Dialize the enzyme during 24 hours at 4 ºC against 500 mL of “tampão de fosfato de potássio
0,02 M, pH 7,4, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M”. Change two
times the buffer solution during the 24 hours.
(cpu translation from italian)
Being a period of plant harvesting, I dedicated myself to the browning and after a period of intake, I found headaches that arises about two hours after they lay down.
Having already experienced some illnesses originating from the Mao inhibitors, I can report that those of the browning would seem selective. They probably interact with mao-b enzymes.
The problem is that by reading on the web what substances act as imao, you wonder why they are products far from feeding. I explain better: our body contains enzymes that oxidize amines and later with age, our body would produce more - perhaps because the amines deflect the mind a bit.
I wonder if drugs acting as inhibitors lead to well-being or not. The periods that you find yourself with inactive enzymes are marked by a general malaise that does not give a glimpse of their pharmaceutical use: headaches, exhaustion, general malaise are the order of the day.
Probably someone will be able to describe more completely their physical and soul state, if they take some psychopharmaceuticals, drugs or constantly use imao on an experimental basis.
From my point of view, you can perhaps notice the increase in vitality for an early period. Once the amines can act without being oxidized, they perform on the body and the psyche as a feeling of stillness that turns into anger if stressed; brief moments of euphoria; non-elastic body - muscle stiffness; disinterest and more.
On some sites I have found various discrepancies regarding the functions of mao enzymes, which leads to the question of whether using substances that make them ineffective is correct. By that I mean to question the constant use, not the sporadic one (from what I have been able to ascertain, to return to the 'normal' situation, it takes months and not weeks).
For example, with well-active enzymes, tyrmine acts as an invigorating one. Serotonin, however, works by giving happiness, if of genuine origin - that is, derived from precursors. The dmt is still active, even if it is a matter of wanting.
Is there anyone who wants to 'disassemble' the aspects of something still little known?
MAOi lead to an indirect increase in amines in the brain whether they are endogenous or external.
Our body tends to rebalance an excess of these amines, ES. serotonin, so when the substance finishes its effect often the balance of these amines is negative and so you have all the various symptoms like headaches, etc...
MAOi are very safe, whether they are of natural origin or of synthesis, problems arise when they are paired with releaser substances such as amphetamines always with regard to serotonin.
In this case the "extra" neurotransmitters cannot be removed from the body because the enzymes are blocked and therefore the effect is extremely more powerful and more durable, but also the risk of serotonin syndrome increases considerably.
Different is the case of DMT and all psychedelic triptammines in fact these molecules are amines and if the enzymes that should oxidize and destroy them are inhibited not only neurotransmitters can not be degraded but the molecule itself remains active for longer.
Extraction of MAO:
https://www.sciencedirect.com/science/a ... 2810000069
,then:
MONOAMINE OXIDASE EXTRACTION
Thaw the samples and centrifuge at 5000 rpm for 10 minutes at 4 ºC.
Wash the mitochondria with “cloreto de potássio 1,15%”.
Centrifuge at 5000 rpm for 10 minutes at 4 ºC.
Wash the purified mitochondria with “tampão de fosfato de potássio 0,1 M, pH 8,2, com Triton
X-100 0,5%”.
Centrifuge during 20 minutes at 10 000 rpm (4 ºC).
Suspend and homogenize the washed mitochondria with “tampão de fosfato de potássio 0,1 M,
pH 8,2, com Triton X-100 1%, colato de sódio 0,5% e -mercaptoetanol 0,002 M” (use an ice
bucket).
Assay for monoamine oxidase and compare with the spectrum from Figure 1.
Maintain the extract with gentle stirring in the cold for 4 hours.
Freeze the samples at – 20 ºC.
ION EXCHANGE CHROMATOGRAPHY
• Packing the column:
Put in the bottom of the column a piece of cotton and a circular net to retain the gel.
Dilute 30 mL of DEAE-Sephacel with 10 mL of equilibrium buffer (“tampão de fosfato de
potássio 0,01 M, pH 7,4, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M”). Put the
slurry of gel in the column slowly.
Start the flow; 1 to 2 mL/min is sufficient (1 drop every 2 seconds)
Note: the slurry of gel cannot be exposed to air!
The column should be equilibrated with 50 mL of equilibrium buffer (“tampão de fosfato de
potássio 0,01 M, pH 7,4, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M”).
• Enzyme dilution:
Dilute the enzyme to about one-and-a-half volume with “tampão de fosfato de potássio 0,01 M,
pH 7,6, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M” (25 mL of enzyme to
37,5 mL of buffer).
• Chromatography:
Elute 10 mL of “tampão de fosfato de potássio 0,02 M, pH 7,6, contendo Triton X-100 a 0,1% e
-mercaptoetanol a 0,002 M”.
Elute 4 mL of the diluted enzyme by employing a sodium chloride gradient (0.0 to 0.5 M) in
“tampão de fosfato de potássio 0,02 M, pH 7,6, contendo Triton X-100 a 0,1% e -
mercaptoetanol a 0,002 M”.
Collect manually 4 mL of eluted enzyme in separate tubes at a flow rate of 1 mL/min.
Read the content of the different tubes at 280 nm. Gather the fractions with higher absorbance.
Wash the column with 10 mL of “tampão de fosfato de potássio 0,02 M, pH 7,6, contendo Triton
X-100 a 0,1% e -mercaptoetanol a 0,002 M”.
Repeat the procedure until all the volume of the dilute enzyme is eluted.
• Ammonium sulfate fractionation
Measure the total volume of the combined fractions and determine the amount of ammonium
sulfate needed for 45% ammonium sulfate saturation (Table I).
Centrifuge at 10 000 rpm, during 25 minutes at 4 ºC.
Dissolve the enzyme in 3 mL of “tampão de fosfato de potássio 0,02 M, pH 7,4, contendo Triton
X-100 a 0,1% e -mercaptoetanol a 0,002 M”.
• Dialysis
Dialize the enzyme during 24 hours at 4 ºC against 500 mL of “tampão de fosfato de potássio
0,02 M, pH 7,4, contendo Triton X-100 a 0,1% e -mercaptoetanol a 0,002 M”. Change two
times the buffer solution during the 24 hours.
(cpu translation from italian)
