zule

Xhol/EcoRI is a restriction enzyme which recognizes the DNA sequence 5’… CTCGAG…3’ and cuts this sequence between the T and the C. Taking into account that the DNA strand is double, the plasmid will become lineal, doble-stranded, with single strand ends: 3'...AGCT...5' and 5'...TCGA ...3' So, we can use the primer TCGA and AGTC to amplify starting with those free ends. It would be much more easily understable with drawings.

Do you want the sequence of the primers for amplifying the fragments of the plasmid by PCR or what exactly do you want? Is this homework? Because such a specific question is not usually something that you “wonder” while you are having a beer

I wrote this article for a BDSM Spanish magazine, CUADERNOS DE BDSM http://cuadernosbdsm.googlepages.com/CBDSM6.pdf . I haven’t still finished translating it into English but it's only a little left and, as it is quite long, I have thought to already post about half of it and when I have finished the translation, I will post the remainder. I would appreciate your points of view in the subject. Any language corrections will also be welcome. As the article was not directed to a scientific public, I preferred not to indicate numbers of reference in the text for not making more complicated its reading. The whole bibliography is at the end of the article; however, if anyone wants references about a particular topic, they only have to ask for them. BIOCHEMISTRY OF THE SUBMISSIVENESS By zule When I was invited by CUADERNOS DE BDSM to write this article, the purpose was explaining the biochemistry of the pain. But I am afraid I have mainly gotten to generate new doubts instead giving explanations. Although it can be found a relatively high number of BDSM psychological or psychiatric papers in the scientific literature, BDSM has not been practically studied from a biochemical or physiological point of view. In fact, even the biochemical works centered in the pleasure study are few. It seems that hedonism has not been given importance in neurobiology. So, the only thing that I could do to write this paper was extracting ideas from studies which only share closeness to the BDSM topic: some of them deal with the pain, a little with the pleasure, and others don’t even deal with any of two subjects. From these ideas, I have tried to establish hypotheses. I want to make clear that a lot of which I have written here are only hypotheses. Although these hypotheses have a scientific basis, I don’t claim them as immovable truths. For that, I would be delighted if anyone wants to discuss these hypotheses for trying to get close truths about the biochemistry of the BDSM. FEAR It is known that different sensations, both pleasurable and painful ones, prompt a first answer which is unspecific and similar. For example, both fear and sexual arousal, cause adrenaline release. Adrenaline makes the body ready for fly, fight, or freeze. You can check that both fear and sexual arousal cause an increase in the cardiac pace, which is due to adrenaline. Therefore, the fear can probably help to improve a subsequent sexual arousal. In a BDSM relationship, the sub knows that the Dom worries about their integrity. Then, why that sub is reacting with fear if they actually know that their Dom is going to look after them? The amygdala is a part of the brain which is mostly activated by fear. Experiments made in relation with phobias, have shown that the amygdala answers fear before the person is conscious of that fear. Because of that, if somebody has a phobia to snakes, although they clearly know certain snake doesn’t have any danger, they are going to feel terrified by it. The phobic can suffer an anxiety state because the amygdala “thinks” there is a real danger and it is going to activate the release of big amounts of adrenaline. The brain cortex also reacts towards fear, but it does it after the unspecific mechanism started by the amygdala has begun. The brain cortex is the part of the brain that allows us to be conscious about what is happening. We will realize we are seeing a snake; or our Master is thinking about what evil to do to us; or we are feeling aroused by a person. And now, this differentiated conscious sensation is going to interact with the undifferentiated sensation coming from other areas of the brain (like the amygdala) to get what we really feel. On this way, during a scene, it might be that our body takes advantage of the adrenaline produced by the fear to increase the pleasurable sensations of sex, surrender, affection… The fact that the amygdala seems to be involved in unconscious sexual desire, besides being involved in the fear, adds more reasons for the fear being able to lead to pleasure. Several studies seem to indicate this involvement of the amygdala in sexual desire. Besides, we could say that the amygdala is a little “disorganized” organ: there is not a clear distinction between the area in the amygdala that is in charge of fear and the ones which are supposed to do other tasks. Even in an individual an area of the amygdala seems to answer to fear, whereas in another individual this same area answers to a different sensation. Summing up, unconscious fear would cause an increase of adrenaline. The amygdala associates fear to excitement. Then, we would realize trough the brain cortex that we shouldn’t be afraid; but that couldn’t stop yet the adrenaline release process. This adrenaline would be added to the one release because of our sexual arousal, love, surrender, etc. In this way, the pleasurable feelings would increase its intensity by using the adrenaline produced by this irrational fear. PAIN The more widespread biochemical explanation about the pleasure caused by the pain is that endorphins released because of the pain would help to prompt an orgasm. However, studies have been made and it has been found an increase in the release of endorphins neither before, nor during the orgasm. There is an increase in the release of endorphins after the orgasm, which would explain the relax feeling subsequent to the coitus. But endorphins seem to make the orgasm difficult, so the pain would reduce the probability to get it. Endorphins due to the pain would cause pleasure, but not sexual pleasure. On the other hand, endorphins are produced during the sexual attraction. It has been also observed that by putting together submissive and dominant animals, in the submissive ones there is a significant increase in the release of endorphins. For this happening, there is no necessary that the dominant animals do anything to the submissive ones; the only fact of being together prompts this increase in submissive animals, whereas the level of endorphins in dominants animals doesn’t seem to vary. During the orgasm there is a significant increase of dopamine; actually the dopamine is perhaps the chemical of the sexual pleasure. Endorphins also cause the release of dopamine, so, here, we may have a pain-pleasure connection: although the endorphins would make getting the orgasm more difficult, the pleasure with or without orgasm would be greater. But now, we have a new problem with this hypothesis: these days it isn’t clear if dopamine is actually the “pleasure neurotransmitter” or it is only the “desire neurotransmitter”. The production of dopamine during a pleasurable situation leads to want to repeat that situation. However, it is not clear if dopamine also is the cause of the pleasure. But then, what substance would be the cause of sexual pleasure??????? But it seems clear that dopamine lead to want to repeat the behavior which allowed its release. This agrees with a typical psychological explanation about the production of pleasure through pain: perhaps in a moment of their childhood, a person experienced pain and a great pleasure at the same time. A large amount of dopamine would be released because of that pleasure, and in our mind would remain recorded a connection between the pain and the pleasure. Finally, other studies seem to demonstrate that dopamine wouldn’t be only involved in pleasure production (or at least in the desire of repeating that situation which caused the pleasure); but it could also be the cause of the unpleasant psychological feeling which the pain induces in “standard conditions”. I am not referring to the physical pain itself, but to the unpleasant feeling which cause that pain. You perhaps wonder how the same substance could cause unpleasantness and pleasure. Neurotransmitters carry out its effect by joining to receptors, a kind of proteins. A neurotransmitter activates certain receptors, and in this way it is produced the effect of that neurotransmitter. But one only kind of neurotransmitter can activate different kinds of receptors. It seems that the activation of some dopamine receptors is related to pleasure, whereas their merging to other kinds of receptor causes unpleasantness. The result could depend on whether dopamine was released in areas where most of the cells had dopamine receptors which activate pleasure or unpleasant ruts. The hypothesis would be that masochistic people had more receptors for the pleasure than for the pain. Besides, the masochists would have to have neurons with mostly receptors for the pleasure very close to other with mostly receptors for the displeasure. In this way, our body could be cheated: part of the dopamine which would have to go to the displeasure centers, would join to receptors leading to pleasure.

It is called NFX1 in eukaryotics.

I hadn’t found this thread when I opened the related http://www.scienceforums.net/forum/showthread.php?t=29536 one (BDSM and Biochemistry). I want to go back to its post lately, but first, I wanted to answer to shaniquaharris in the hope that he still reads this: I strongly believe you are into the BDSM. There are a lot of people who have felt they are not normal because of those feelings, including me. But the BDSM is merely a way of feeling which a lot of people share to a greater o lesser measure. I also understand your feeling what you have seeing in websites about BDSM is fake, because much of that you see on BDSM websites is fake (generally the more commercial the web, the more fake the situations), and people who really have deep BDSM feelings can feel disappointed. I would advise you to better visit BDSM forums, where people can share their feelings. I don’t know where you are from, but there is an international social web, http://www.fetlife.com where there are a wide range of ways of feeling the BDSM and fetishism, from merely a from-time-to-time role-play to real rooted feelings. I think it must be the bigger social web in the world about BDSM, I belong to it and it has hundreds of thousands of members and hundreds of different groups. But you can also look for other social webs or forums when you can feel well. The important thing is that you know you are normal, what you have is a gift to enjoy. Looking forward to hearing from you. zule Merged post follows: Consecutive posts mergedagentchange’s ideas in the subject are strongly widespread. I myself though in the past I was insane because of my desires. But ours is really only another way of living the sex and the relatioships. I wanted to answer to ParanoiA because there is a lot of discussion about this topic in the BDSM environment. I think you can say there are BDSM people in the same way you can say there are homosexual people. That doesn’t mean there are not levels. There are bisexual people, who could be more biased to homosexuality or heterosexuality and there are people who is mostly heterosexual but are curious about homosexuality and try it. It’s the same with BDSM. There are people whose way of feeling the sex and the relationships is almost exclusively through the BDSM, while other people only use the BDSM to spicy the things up. I consider myself a BDSM person; although I wouldn’t have an entire BDSM lifestyle, what it is called a 24/7 relationship, I need the BDSM to have a full relationship. I wrote an article about “Biochemistry of Submissiveness” for a BDSM Spanish magazine and I started to translate it to English for a forum in the social web I have previously referred. I am uploading it by parts in that forum but, as I have almost finished the translation, when it is complete, I will upload it here in the threat “BDSM and Biochemistry”

Chemicals interactions allow to have thoughts; however, thoughts are not chemicals, but abstracts entities. So, for me, it doesn’t make sense to talk about the dimensions of the thoughts.

It is all about energy transformations. Electron transference liberates energy, because of the difference in the Gibbs’energy between the different complexes in the ETC. This liberated energy is used to create a proton gradient: in a side of the membrane the concentration of protons is higher than in the other. This difference of concentration means a potential energy: when the concentrations on both sides of the membrane are allowed to level, this potential energy is liberated and used by ATP synthasa to synthesise ATP. The potential energy of the proton gradient is similar to the potential energy of the height: we need energy to rise an object, but then, we get energy when we let that object fall.

I like this video http://www.cnr.vt.edu/DENDRO/forestbiology/photosynthesis.swf But if you want to have a wide knowledge of the photosynthesis, I think you could find it better at books you surely have in the library of your college.

Genecks: I think the splinter wouldn't be broken down; it would be encapsulated and/or expelled from the body.

I don’t think we will never get to create a being as us. For creating the brain of a being as us, we only have our own brain, and I believe that to copy something, we need a much more complicated tool than the thing we copy. So, I believe that, if the world still exists for a very long time, perhaps evolved humans with one much more powerful brain than ours could be able to create a being as us, but not a being as them.

I think you are wrong: If you crossbreed a cream-colored horse with a black one, you will get a brown colt, the same which occur in humans. If you want a black with white spots colt, the horse or/and the mare parents will have to be spotty. The difference is that we don’t have spotted human father or mother in order to get a spotty baby. As far as I know, the only animal where you can get a spotty cub from two plain parents is the cat, as GDG has explained.

Both, fungi and animals, need organic matter to survive, whereas plants nourish from inorganic matter. This is because only plants can execute the photosynthesis.

I don’t actually have any reference. My post came from a course I took about biochemistry of drugs. But now, I have gone over my notes from that course and I have read that if the use of marihuana is occasional, the THC is eliminated in a week, but in chronic users it can accumulate in adipose for about two months. So, you are right. I have tried to look for any reference in the pubmed but I only find time limits of detection, which depend on the measurement method. Anyway, two months is not a long time for consider withdrawing the marihuana a good treatment to figh against the problems it could cause in certain people, not needing any additional one.

No, THC is completely eliminated from your body in about a week. The only place you can detect it for a long time is in your hair, but your hair is dead, the THC doesn't go back to your blood from there.

In my teens and first youth, I took too too too … much recreational drugs: I ate LSD as if they were candies; I took a lot of speed and cocaine; of course marihuana and alcohol; and occasionally heroine, amphetamines and everything that turned up in my hands. This was mainly between 14 and 25 years old and I haven’t taken any illegal drug since before I was 30. I even gave up smocking 5 years ago. So, I can say that I have healthy habits if I compare them with those I had before. Still, I have too much carbohydrates, I am addicted to them, but when I see what most of people usually eats in certain countries, I realise my mostly Mediterranean diet is quiet healthy. Regarding alcohol, I don’t drink often, but when I drink, I usually actually drink. But the chairman of the “Grupo Español de Neurotransmisores (GEN)” always repeats that Spanish red wine is neuroprotector. And as the remainder of the members don’t want to be excessively nationalist; we also have Scotch whisky, Cuban run, American bourbon, Russian vodzka… I send you some photos from a meeting closing ceremony, so you can see how we are neuroprotected. I am the blonde with the blue shirt in the last photo.

Perhaps is only questions of semantics, iNow, but I am not agree with your statement about not being a place where the memory is stored. It seems to be stored in the neocortex, but it is true that it is not stored as if the brain was a DVD. The form of storage would be the circuits which are formed due to the new synapses constituted by synaptic plasticity. The activation of these new built circuits would allow remembering. Regarding the first link, it doesn’t actually talk about storing memories, but about encoding them. The memories first have to be encoded in the hippocampus in a process in which neurotransmitters dopamine and acetylcholine seem to be mainly involved. Then, memories are progressively transferred to the neocortex to be stored. Thank you very much for the second link. It seems amusing. I will look at it carefully. I don’t know how you do to find always the best links.

Atoms are arranged in molecules and these are in turn arranged in macromolecules and so on successively in more complex structures in a way they have the minimum energy. So, it is necessary a big amount of energy to destroy all these structures and so get the atoms free to recombine in other way, which always will be the one of minimum energy again at the end. Any way, I think that this post should be in “Physics”, where I am sure somebody would explain it much better.

I got that idea mainly for my job as a researcher in neurochemistry. But there is a high probability that I am wrong, because I am blonde When someone is talking about connections between neurons, at least they specify other thing; they are talking about synaptic junctions. Glutamate is the main neurotransmitter implicated in memory formation. The activation of glutamate receptors prompts a signalization cascade that will lead to “short time memory” (seconds or minutes). For “long term memory”, we need repetitive stimulations of glutamate receptors. These stimulations lead to activation or inhibition of genes, prompting synaptic plasticity: molecular changes are going to happen, being the more notable the variation in the number and distribution of connections between the different neurons. The more accepted theory about the long term memory is that memory is stored by means of changes in the connexions between the different neurons

Simplifying, memory is stored by means of changes in the connexions between the different neurons.

Who told you your problems might be due to masturbation???????

There are of course genes which predispose to nicotine addiction; those ones are related with nicotine receptors and with dopamine. But nicotine receptors have not evolved in our body with the purpose to interact with the nicotine of the tobacco. They have evolved to interact with our own neurotransmitter acetylcholine and so to carry out its function. On the other hand, the tobacco plant has found an evolutive advantage imitating the effect of acetylcholine in its interaction with nicotinic receptors. As DH has indicated Europeans have been in contact with tobacco for about 20 generations, while the acetylcholine has acted on nicotinic receptors for millions of years… Regarding dopamine, this neurotransmitter and its receptors are implicated in the addiction to all drugs, because it’s the neurotransmitter of the reward system. Genetic variations in this neurotransmitter or its receptors will lead to different tendencies to drug addiction.

As iNow has indicated, during the birth there is a great increase of oxytocin in the mother’s blood. The oxytocin prompts the desire of protection, so the mother accept an offspring although it isn’t its. When levels of oxytocin have decreased, the mother has probably habituated to its new offspring.

I was tired of so hot it was being in Madrid. So, I have spent 15 days in England (Bath, London and Brignton). I have got tired of so cold it was there, but English people is nicer than it is said about them.

Welcome, Victoria. I don’t think you must apologize for being you first post. I would swear that every member who has written on this forum has had a first post When insulin joins to its receptor on the cell, prompts a signal cascade. This leads, not only to the activation of the glucose transporters that insert glucose into the cell, but also to other effects. Those effects are mainly focused in increasing the synthesis of metabolites (glycogen, fatty acids) and reduce their degradation. When we have carbohydrates, those are degraded and the glucose coming from this degradation activates the release of insulin. So, in a healthy body, the glucose is going to be taken by the cells due to the releasing of insulin. But if more insulin is added as you say, too much glucose is going to be taken and transformed into glycogen and fatty acids. Due to the fact that brain, in physiological conditions, uses only glucose for working, a little time after you have eaten, your brain is going to ask for more carbohydrates: You are going to have hungry and your brain is not going to work properly until the levels of insulin decrease enough. But this would be the case if you inject the insulin. However, you were talking about “drinking” the insulin. The insulin is a protein which is going to be degraded into amino acids in the digestive system before passing to the blood. So, I suppose that nothing is going to happen if you drink insulin.

I coudn't express better my own opinion