Hi Petra, Salimetrics do an assay for hair cortisol, which is very good. It is perfect for examining chronic stress, (provided that the subjects don't have military hairstyles of course) such as over a period of months. In our opinion at IPRO, they offer some of the best commercial assays on the market and this was confirmed in Psychoneuroendocrinology (2013) 38 50-57 where their salivary assay correlated very well with LC/MS. I believe that their hair method is very good also.

Cortisol can be measured in blood, saliva or hair. There is reasonable agreement between blood and saliva measures (although the absolute blood values are higher). If you like a simple non-invasive point-of-care test for saliva, we have one for cortisol. We commonly use it in sport as a measure of training status, but we also preform cortisol tests frequently for our corporate clients. In both of these applied settings the cortisol is often expressed as a ratio with testosterone or DHEA-s respectively. One of the problems is that cortisol measurements vary greatly as a function of time of day, which is why interpretation can be tricky. It is not even just time of day, but time since awakening that needs to be monitored for effective analysis. Our corporate clients often take several measurements throughout the day, as the shape of the profile is very useful and gives a better picture of stress than a simple one-off measurement.

Joe

([email protected])

Thank you for your answers for now. As the design of the experiment is not ready yet I am not able to provide any more information. The participants of the experiment will definitely be humans.

There is a lot of literature out there on cortisol measurement in stress, and there are many pitfalls in measuring cortisol.

This is not meant to be an exhaustive guide, but from memory you have to deal with

1. The natural circadian rhythm with the morning cort rise (the CAR) and the afternoon nadir. Most "stress tests" are done in the afternoon for this reason.

2. Free vs total cort? in plasma, mostly total cort is measured, but in saliva it is free cort? what do you want to measure and why?

3. cort levels saturate CBG (the main cort binding protein) in the morning, and often a lot of free bioactive cort is seen, wheras in the afternoon, Cort is usually below saturation, so that an increase in plasma total cort is not necessarily seen in saliva as free cort.

As a general rule saliva is the best for beginners. Take as many samples as you can, if you do things like the CAR do repeated morning with multiple (as many as possible) time points per CAR, and if possible do all your testing in the afternoon.

There are many good stress researchers in Germany, including Dirk Hellhammer, Brigitte Kudielka, Stefan Wust to name just a few, and any one of them I am sure would be delighted to help you.

Hope this helps

Jon

Hello Jon, thank you for your helpful answer. It is thought to be an instrument of control, because I want to offer stress course and would like to control the stress level besides. But I have not planned any details.

We use saliva to measure cortisol, but I do agree, that regardless of which measure you choose, circadian variation should be factored in. It will also be important to try and ensure that your subjects are reasonably equivalent regarding usual levels of stress experienced. A flattened amplitude of cortisol circadian variation, low morning cortisol levels or excessive cortisol levels can all result from ongoing stress (and thus can cloud group results). It would also be worth using a pen and paper test to test for Morningness and Eveningness, as the 3 categories (morning/evening/neither) will show different circadian profiles. As neither types are the most common, they would be the best to use. Cortisol is quite variable as a measure and so good experimental controls are really necessary.

You can measure it from saliva more about that in my thesis

Dr Pohler, do you have a summary in English?

Gudrun, if you want to measure acute stress, we have two biomarkers that are much quicker to respond on our real-time device, namely alpha-amylase and secretory IgA. Our real-time device offers quicker and cheaper alternatives to standard Lab assays and can analyse small numbers of samples if need be, at the same unit cost, unlike ELISA that really needs 40 samples at a time to be remotely cost-effective.

The whole thesis is in english, just take a look

Thanks Dr Pohler

Hi Joe,

We also use s-IgA but find it quite variable. Do you have a paper that I can have a look at? You're clearly doing something better than we are to be able to use s-IgA as a biomarker

Thank you for your answers. The programme is growing and step by step it will be going into practice. Then I will find out whether it is a good idea to make stress tests as well.

We usually ure saliva samples too (Salivettes). You must use it carefully and (following some protocols about it´s use and conservation), but they provide good results in cortisol collection.

Good luck with your work!

Our Point of Care salivary cortisol test is now on the market for those interested. It is mainly used in sporting and corporate applications, but would be useful for pilot studies etc in the academic world. For full-blown research, where data is not time sensitive and large numbers are involved you may as well use standard lab assays. For those in Australia, this test is available from Stratech Scientific.

Thank you for your answers. I found another very interesting thread also dealing with this subject: https://www.researchgate.net/post/Which_factors_affect_salivary_cortisol_concentrations?cp=re65_x_p3&ch=reg&loginT=VxvBRpLt_WZJwP2F9Ylshhpa9O3vVqCSf6l9xQT8fpc%2C&pli=1

Hi Petra, Salimetrics do an assay for hair cortisol, which is very good. It is perfect for examining chronic stress, (provided that the subjects don't have military hairstyles of course) such as over a period of months. In our opinion at IPRO, they offer some of the best commercial assays on the market and this was confirmed in Psychoneuroendocrinology (2013) 38 50-57 where their salivary assay correlated very well with LC/MS. I believe that their hair method is very good also.

Hi Petra,

And this method (hair) even lets you to evaluate temporal change of cortisol using different lengths of hair. There are papers about it, I can check my endnote and send you if you are interested in.

Dear Nese, would you please send the paper to me, too?

When one tests stress in hairs one could also test drugs and alcohol for the last three months.

As both measurements have good correlation, the election of one or other, I think, should be related with what you want to study.

• If what you want to know is to assess how the patient diary stress level is, for to see if is during at the work time or even is persistent after that, you should choose the analysis of saliva and do a curve starting at 8 am. since 12 pm., one sample each 4 hours.The best, I think, it would be make this curve one time in several weeks for a more accurate assessment. Also could be used for Cushing's syndrome, because you take sample at late at night.
• If you only want to know if the patient has been subjected to a chronic stress for a period of time, you should do the analysis of hair, because it will be the result of the accumulation of this steroid hormone in the hair not a punctual level like in saliva occurs.

For chronic stress, It could be of interest to do a preliminary study in saliva for to characterize the patient and after, go doing the follow up with measurements in hair each xx period of time.

Dear Dr.  ;

         I suspect that you will find interesting the article below, which discusses the function of cortisol in the body as a hormone that inversely mobilizes the body against potassium wasting intestinal diseases, and in http://charles_w.tripod.com/cortisol.html   with the references and links. If you see anything that can be improved please let me know. You may see a journal article that discusses this briefly in the 1998 vol. 51 issue of Medical Hypotheses, p 289-292.

            Sincerely, Charles Weber

THE PURPOSE OF CORTISOL and CORTICOSTERONE

by Charles Weber --- If you use information in this article, get a second opinion from a doctor.

ABSTRACT

It is proposed that the primary purpose of the glucocorticoids, including cortisol (hydrocortisone), is to mobilize the body to resist infection. They do so by normally altering processes which increase pathogens' growth or their adverse effects and then declining when under attack. Cortisol is for intestinal disease (diarrhea) and corticosterone serum disease. Glucocorticoid mobilization for fight or flight is an adjunct, made possible because most processes which resist infection impair fight or flight. A different hormone controls those which do not.

Potassium loss is the most serious aspect of intestinal diseases, so the electrolyte capabilities of cortisol, but not corticosterone, are oriented toward conserving potassium. Low cell potassium reduces adrenal synthesis of cortisol, but not corticosterone. Sodium, water, glucose, amino acids, chloride, hydrogen ion, copper , and numerous others are controlled by cortisol in such a way as to survive during intestinal disease.

Some gram negative bacteria have an endotoxin which subverts this strategy by forcing the secretion of huge amounts of ACTH, which is the chief mediator of cortisol. A glucocorticoid response modifying factor (GRMF) and interleukin-1, raises the effective set point of cortisol. The immune cells thus take over their own regulation, using interleukin-1 to mediate production of cortisol via ACTH.

Scroll down to INTRODUCTION below.

CONTENTS of other chapters Back to INTRODUCTION chapter -- II. Arthritis Research -- III. Arthritis and Potassium -- IV. Roles of Potassium in the Body -- V. Electrolyte regulation (sodium and potassium) -- VI. Purpose of cortisol -- VII. Copper nutrition and physiology -- VIII. Nutritional Requirements -- IX. Potassium in Foods -- X. Processing Losses -- X,cont. Losses in the kitchen -- XI. Supplementation -- Side Effects and Heart Disease -- When Potassium in Blood is too High

For unknown words see this site and for definitions click on "find terms".

INTRODUCTION

This paper will propose that the primary purpose of cortisol (hydrocortisone) and corticosterone in mammals is to mobilize the body's physiological processes against infection and its adverse effects, cortisol against potassium wasting intestinal disease and corticosterone against serum disease. These steroids control a large number of enzymes, hormones, and processes, most of which could enhance growth of pathogens or make the adverse symptoms worse. The few which do not, do not affect immunity either, and are probably opportunistic adaptations of these hormones to peripheral functions. Extinction of juvenile play traits is an example.

Glucocorticoids mobilize immunity by declining their serum concentration. This inverse style is highly desirable, otherwise a pathogen could easily overwhelm the immunity defenses simply by evolving an enzyme which could degrade steroids. Some circumstances controlled inversely enhance an animal's survival from the adverse effects of bacterial poisons or the animal's own defenses. Such a defense would be control of blood pressure. This control, I suspect, is largely to protect infection damaged and copper starved blood vessels from hemorrhage.

Please keep in mind as you read this, that cortisol's functions to inhibit or stimulate become the reverse, to stimulate or inhibit, upon decline respectively. This concept will be handled by use of the phrases "inversely stimulates" or "inversely inhibits" respectively as cortisol declines.

Cortisol is controlled by the pituitary peptide ACTH. 1 ACTH is in turn controlled by the hypothalamic peptide, corticotrophin releasing factor [CRF], 2 under nervous control. CRF is synergistic with arginine vasopressin, angiotensin II, and epinephrine. 2 Therefore ACTH and CRF cannot be overwhelmed by bacterial degradation either. ACTH probably controls cortisol by controlling movement of calcium into the cortisol secreting target cells. 1

Cortisol prevents proliferation of T-cells by rendering the interleukin-2 producer T-cells unresponsive to interleukin-1 (IL-1), and unable to produce the T-cell growth factor. 4 That cortisol often increases during infection does not make this hypothesis invalid because when activated macrophages start to secrete IL-1, which synergistically with CRF increase ACTH, 5 T-cells also secrete glucosteroid response modifying factor [GRMF] as well as IL-1, both of which increase the amount of cortisol required to inhibit almost all the immune cells6 (there is no information in the literature that indicates whether GRMF is the same as transcortin, also called cortisol binding globulin, or not). Thus immune cells take over their own regulation, but at a higher set point. Even so, the rise of cortisol in diarrheic calves is minimal over healthy calves and drops below with time. 7 The cells do not lose all of the fight or flight override because of interleukin-1's synergism with CRF. Cortisol even has a negative feedback effect on interleukin-1 5 which must be especially useful against those diseases which gain an advantage by forcing the hypothalamus to secrete CRF, such as the endotoxin bacteria to be discussed later.

The suppressor cells are not affected by GRMFs, 6 so that the effective set point for the immune cells may be even higher than the set point for physiological processes. It may be that the GRMFs have a different spectrum of effects for each of the physiological processes in order to fine tune the immune response in order to optimize the attack against different organisms.

It seems to me that resources diverted to immunity or denied to non-viral pathogens usually diminish an animal’s performance when fighting or fleeing. Therefore, the cortisol system can be overridden by perceived danger. This is no doubt made desirable because it takes several hours or more for pathogens to rise to dangerous levels, but only a few seconds for a predator to kill an animal. Anxiety is also factored in because, I suggest, cortisol operates by changing the nucleus commands to send RNA for production of enzymes, etc. in almost every case and the various diffusion steps take an hour or more to complete. Therefore, an anticipation of danger would be very desirable.

The desirability of inhibiting activity during infection is no doubt the reason why cortisol is responsible for creating euphoria, 8, p.736 as does aldosterone, 9 and presumably the reverse upon declining. The desirability of not disturbing tissues weakened by infection or of not cutting off their blood supply could explain the inverse stimulation of pain widely observed for cortisol. These neural mechanisms as geared to stress have been emphasized in concepts concerning glucocorticoids as pioneered by Selye up to now. Nevertheless, when a process must move in the same direction for both immunity and fight or flight, a different hormone system controls it for stress. An example is release of ceruloplasmin by the liver, which is controlled for purposes of stress by epinephrine and by an unknown hormone for immunity to be discussed later.

The most dangerous digestive diseases produce a protein poison which stimulates cyclic adenosine monophosphate [c-AMP] hormone in such a way that the intestines cannot remove water from their contents10 and thus cause diarrhea. Since potassium in food and the 2.5 grams or so secreted with digestive fluids can only move into the blood stream passively, 11 this causes a large loss of potassium. Judging by the reduction of the death rate in babies with virulent diarrhea from 34% to 6% by potassium supplements12 in spite of the danger of hyperkalemia (high serum potassium) during dehydration, the loss of potassium implied is the most serious consequence of diarrhea. When this poison first evolved, it must have been catastrophic to terrestrial vertebrates. Even today, after what was probably a major evolutionary transformation of cortisol, the diarrheas are among the most important causes of mortality in the tropics, especially cholera 12a and diarrhea was the third leading cause of death 100 years ago in the USA, ahead of heart disease. Even currently a virus called rotovirus kills 600,000 children each year, most of the deaths in India, sub Sahara Africa, and western Africa 103. It must have been imperative to evolve mechanisms to surmount those pathogens. In most mammals, a wide range of processes are stimulated by cortisol, each of which would make an animal less able to resist potassium and water wasting intestinal disease. Rodents have very little cortisol, which may be related to a marked inhibition of the effect of cholera toxin by rodents' intestinal contents. 13 Also, c-AMP increases water absorption in their ascending colon, opposite to the effect in their descending colon. 14 This makes rodents dubious for experiments on the hypothalamic-adrenal axis and perhaps for any experiments. These attributes are possibly the reason why rodents have the more vertebrate species than any order 102 and why they are so well adapted to desert life.

DISCUSSION

POTASSIUM

The greatest urgency during diarrhea is to prevent loss of potassium, since there is no storage of potassium in any cell. In cells, 88% of the potassium is in free solution. 11 Indeed, one of cortisol's functions conserves potassium. It has been suggested that cortisol tends to move potassium inversely into the cells [cortisone]. 15 If this is the case, potassium is inversely conserved by lower secretion of cortisol (dexamethasone). 16 In order for potassium to move into the cell, cortisol inversely moves out an equal number of sodium ions. 15 It can be seen that this should make pH regulation much easier, unlike the normal potassium deficiency situation in which about 2 sodium ions move in for each 3 potassium ions that move out17, p.445 which is closer to the DOC effect. 15 This is probably the reason why the cell becomes acid during a deficiency caused by low potassium intake. 18 Nevertheless, cortisol consistently causes alkalosis of the serum [inversely acidosis] while in a deficiency pH does not change. I suspect that this is for the purpose of bringing serum pH to a value most optimum for some of the immune enzymes.

Potassium is also inversely inhibited from loss in the kidneys somewhat by cortisol [9 alpha fluorohydrocortisone]. 19 Potassium is primarily blocked from loss in the kidneys by a drastic decline of aldosterone during dehydration. 20 Aldosterone acts on the last part of the kidney tubules and the lower colon. 21 In the colon, aldosterone reverses the normal inward flow of potassium, or at least stops its reabsorption22 and so inversely conserves potassium there. Aldosterone is directly controlled by potassium and inversely by osmotic pressure20 while angiotensin II is required. Thus as osmotic pressure rises during dehydration; aldosterone undergoes a drastic decline. Aldosterone also backs up cortisol by possibly inversely moving potassium into muscle cells somewhat. 22

To be useful in combating a potassium wasting disease, it would be necessary for cortisol to decline at such a time. A high potassium media, which stimulates aldosterone secretion in vitro, also stimulates cortisol secretion from the fasciculata zone of dog adrenals. 23 and potassium loading increases ACTH and cortisol in people [101]. Therefore, low potassium should decrease cortisol secretion by the adrenals in vitro in dogs and people. At the same time, potassium has no effect on corticosterone secreted by the adrenal fasciculata. 24 Since the fasciculata accounts for 5/8 of the corticosterone secreted, the net effect is very little decline in corticosterone secretion. This is evidence that the body does not rely on corticosterone against diarrhea. Potassium chloride supplements do not affect cortisol or corticosterone plasma concentrations in humans in vivo when the cell content is adequate. 25 I know of no experiment which would establish the effect of potassium, cholera toxin or detection of intestinal pathogen microbes on ACTH. ACTH has its greatest effect on cortisol and 18hydroxy 11 deoxycorticosterone [18OH DOC] That last hormone affect is probably to acidify the blood during an infection.

SODIUM

Cortisol is used to stimulate sodium inward for fresh water fish and outward for salt-water fish. 26 The necessity of conserving potassium while still unloading electrolytes to maintain osmotic pressure may explain cortisol's inverse sodium losing power in the small intestine in mammals. 27 By using the intestine to excrete sodium, less water is needed for kidney processes, which is crucial during diarrhea. Sodium depletion does not affect cortisol, 28 so cortisol is not used to regulate serum sodium. It is known that the sodium retaining hormone, 18-hydroxy 11-deoxycorticosterone [18OH DOC] acting on the kidneys is strongly dependent on ACTH. When ACTH sinks to zero, 18OH DOC also does. 29 Therefore, it also is inversely involved in unloading sodium in what little water is excreted from the kidneys. The need for sodium chloride by diarrhea bacteria in order to grow rapidly30 may be the main reason why cholera enterotoxin is so successful for this bacterium and of course increased water undoubtedly assists it also. 18OH DOC is probably the hormone which stimulates hydrogen ion excretion, so loss of this excretion would assist acidifying the serum as mentioned above as well.

If my contention that 16-alpha 18-dihydroxy 11-deoxycorticosterone [DOH-DOC] is relied on to excrete excess sodium and to conserve potassium17 is valid, it should follow that ACTH and/or cortisol either have no effect on DOH-DOC or, possibly more usefully, to inversely stimulate it. It should also be desirable for DOH-DOC to exert its effect in the intestines because in nature it is almost always during diarrhea that the body experiences a potassium deficiency and sodium glut. I have no direct evidence for either phenomenon. However, it is known that DOH-DOC has very little affect on the kidneys.17, p.446 The malaise, headache, loss of appetite, insomnia, and muscle cramps created by DOC injections31 may be due to the loss of potassium and retention of sodium, resulting from increased DOC, causing DOH-DOC to rise, since none of these symptoms appear from a high sodium and potassium diet which stimulates DOC.17 Some of those attributes would be useful during diarrhea, but I have no evidence for DOH-DOC's role. 11-deoxycorticosterone [DOC] is the only steroid left of the four I proposed for electrolyte regulation.17 Sodium retention must never completely disappear. This may be why, as possibly the only renal sodium retainer left, DOC has acquired its auxiliary powers with respect to amino acids and copper to be discussed later and why a fall in leucocyte potassium of over 10% is observed from DOC32 and a decline in muscle potassium,33 thus joining cortisol in inversely conserving potassium. It also probably explains why it is mediated partly by ACTH since ACTH must surely largely be an immune hormone with stress as an adjunct.17,p.445

WATER

Cortisol also acts as a water diuretic hormone. Half the intestinal diuresis is so controlled.27 Kidney diuresis is also controlled by cortisol in dogs.34 The decline in water excretion upon decline of cortisol [dexamethasone] in dogs is probably due to inverse stimulation of antidiuretic hormone [ADH or arginine vasopressin] the inverse stimulation of which is not overridden by water loading.34 Humans also use this mechanism35 and other different animal mechanisms operate in the same direction.

Since loss of water is the circumstance which produces the worst adverse effects of diarrhea, it would seem to be logical to use dehydration as a signal to decrease cortisol. Cortisol has been found to vary directly with water intake 36a ACTH hormone production is inhibited by water deprivation at the pituitary level. Base secretion of ACTH is not affected, but high plasma ACTH resulting from immobilization stress is almost cut in half. Base corticosterone is increased in plasma from dehydration, but the much higher corticosterone from immobilization stress is not affected by water status.36 The above is additional evidence that corticosterone is used by the body to fight serum disease and cortisol is used to fight intestinal disease.

GLUCOSE

Reinforcing the concept that cortisol is relied on more for intestinal disease control and corticosterone for serum disease is the circumstance that corticosterone at physiological levels shows a marked inhibition of insulin and enhancement of glucagon in vitro.37 Cortisol shows a small inhibition of glucagon which reverses in a short time and has no affect on insulin.38 Insulin is used to help prevent hyperkalemia [high serum potassium] by the body. As glucose moves into the cell, it takes potassium with it. This mechanism is only used at low potassium intakes. At an intake of 8 grams per day, insulin stays normal.39 This is logical since there is no need to conserve potassium at high intakes and aldosterone is relied on to lower serum potassium. Cortisone greatly inhibits insulin secretion.38 The cortisone-cortisol equilibrium may explain why in vivo experiments contradict the above.40 It is possible that this equilibrium may permit the body to change cortisol glucose responses for particular kinds of situations.

The inversed stimulation of insulin by corticosterone would lower serum glucose and thus deny glucose to pathogens. Such an aptitude in cortisol would be of little value if my thesis is correct, and could even endanger an animal from hypokalemia [low serum potassium] during diarrhea. A sudden withdrawal of glucose by insulin in a potassium deficiency can lower serum potassium enough to be lethal. However, apparently there is an advantage in locking up the potassium that does enter the cell in a more orderly manner with glycogen, because DOC inversely stimulates glycogen formation.41 Cortisol does inversely cause serum glucose to fall, but this is probably an indirect effect caused by inverse inhibition of amino acid degradation.

The intestinal brush border disaccharide enzymes are inversely inhibited by cortisone.42 If it is cortisol that is actually involved, this could be a mechanism to deny energy to bacteria incapable of using sucrose. However, present day cholera can ferment sucrose43, p.557 so it would have to be an attribute developed against diarrheas which evolved before cholera evolved. It is also possible that it helps prevent the hypokalemia above or make copper more available if those enzymes are copper catalyzed. The fact that sucrose and fructose make a copper deficiency much worse suggests that they are. [100]

AMINO ACIDS

Glucocorticoids have the attribute of inversely lowering amino acids in the serum.44, p.273 They do this by inversely stimulating collagen formation, increasing amino acid uptake by muscle, and stimulating protein synthesis.44, p.273 Cortisol also inversely inhibits protein degradation.45, p.207 Such an attribute would help deny amino acids to bacteria. An additional advantage is that collagen can be very useful in repair of infected tissue. An indication of this last is that loss of collagen from skin by cortisol is ten times greater than from all other tissue in the rat.45 Thus the skin can be a reasonably safe source of energy during stress and be rapidly repaired during damage preliminary to or caused by infection. Lowering serum amino acid or even tissue damage repair during intestinal disease should be not nearly so advantageous. An indication that it is not is that DOC acts in the opposite direction for collagen [mice]46 and thus tends to cancel cortisol's effect if the same thing happens in other animals.

It can be seen that denying amino acids to bacteria above could be very advantageous in a serum infection. However, the inverse generalized stimulation of protein synthesis44, p.273 [I'm not certain how generalized it is] could have additional survival rationale against digestive disease. 40% of the protein synthesis is in the intestines of the rat, much of it for synthesis of IgA.47 IgA acts as an inert, nonlethal coating on bacteria to prevent adhesion to intestinal walls47 and is the predominant immunoglobulin in the human intestine.43, p.597 Its production depends on the vitamin A metabolite retinoic acid 105. Mora, et al discus how specialized B cells from lymphoid tissue associated with the intestines migrate to the intestines’ wall where interleukin 5 and 6 peptide hormones permit retinoic acid to stimulate secretion of IgA112, p1160. This is no doubt the reason why vitamin A supplements decrease mortality from diarrhea by 34% in malnourished children112. Cortisol [opticortinol] probably inversely stimulates IgA precursor cells in the intestines of calves.48 Cortisol also inversely stimulates IgA in serum, as it does IgM, but not IgE49, although serum IgA does not depend on vitamin A. I cannot account for the effects on IgM and IgE.

Cortisol has an opposite effect on liver than it has on muscle, but I cannot tie this for sure into the immune concept now. I suspect that it may be to provide a small amount of maintenance amino acids when the muscles are withdrawing them from the blood and possibly also to provide liver amino acids for IgA. That same inability of mine is true of its inverse activation of luteinizing hormone.

HYDROGEN ION

Sodium, potassium, and chloride make strong bases and acid so that any unilateral movement by any of them has considerable implications in hydrogen ion control. Cortisol inversely inhibits gastric acid secretion.50 Since hydrogen ion interferes with potassium excretion at the kidneys,51, p.215 this could be having a potassium conserving effect, especially since gastric secretion carries 0.6 grams of potassium per day into the stomach as well. Corticosterone has a much greater effect on gastric acid secretion than cortisol.50 I cannot explain why it should have any affect at all unless there is some advantage to keeping the serum at a lower pH during infection for enzyme enhancement a possibility already mentioned. Some leucocyte enzymes have a pH optimum lower than serum. If so, 18hydroxy 11 deoxycorticosterone, which reduces bicarbonate and stimulates hydrogen ion excretion at the kidneys,52 operates in the same direction, since it also declines with ACTH half again more than cortisol.53 Cortisol's only direct effect on the hydrogen ion excretion of the kidneys is to inversely inhibit excretion of ammonium ion by inactivation of renal glutaminase enzyme.54 Glutaminase splits ammonia off of the amino acid glutamic acid, and this provides ammonium ion to take the place of potassium for excretion. However, cortisol's presence is necessary for the other hydrogen ion excretion regulator to operate.54 There would have to be some restraint on hydrogen ion loss because when potassium is deficient, the kidneys fail to absorb chloride and the serum tends toward alkalosis.55 Perhaps cortisol's inverse inhibition of gastric secretion being lower than corticosterone's is a compromise made necessary by the advantage in keeping the stomach reasonably acid, below a pH of 6, in order to help prevent reinfection by cholera bacteria.43, p.556 The acidosis of serum that attends cholera43, p.601 may become too high, so this lower inhibition may also be a compromise to help solve such a situation. The net effect of glucocorticoids is to inversely acidify the serum.

CHLORIDE

Chloride is intimately involved with potassium loss because when the cell loses potassium to take the place of serum losses and sodium migrates in, chloride must also be excreted as the only ion which has a chance of maintaining serum pH. In a potassium deficiency chloride is lost.55 This is a serious circumstance in nature because chloride is not bound very well by soils. It is a seriously limiting element inland where vegetation is devoid of it as a rule. Some indication of its importance is that it is the only essential nutrient we can detect and be attracted to other than water [the salty taste].

Net chloride secretion in the intestines is inversely decreased by cortisol in vitro [methylprednisolone].56 Cholera toxin forces chloride secretion to reverse from flow inward to larger flow outward.57 Thus cortisol tends to inversely neutralize cholera's effect. There is no net movement of chloride by cholera toxin in vivo.58 It is possible that this attribute is related to keeping the serum pH low as mentioned above, that is to say, acidic. It is possible that movement of sodium and/or chloride into the intestines is the chief advantage that diarrhea bacteria attempts to gain from their water losing toxin.

COPPER

The immune system is very sensitive to copper availability. Spleen of copper deficient animals show little growth during infections.59, p.334 Even a mild deficiency causes spleen derived immune cells to be significantly less competent as stimulators in general and also to be stimulated by endotoxin, pokeweed, or concanavalin A.60 Resistance to infection is reduced somewhat by a deficiency.59, p.334 A reduction in neutrophils is the first symptom of a deficiency in children.59, p.336

It is therefore probable that increasing copper for immune purposes is the reason why many copper enzymes are inversely inhibited to an extent which is often 50% of their total potential by cortisol.59, p.337 This includes lysyl oxidase, an enzyme which is used to cross link collagen and elastin.59, p.334 DOC acts in the same direction as cortisol for lysyl oxidase.59, p.337 Particularly valuable for immunity is the inverse shutdown of superoxide dismutase by cortisol61 since this copper enzyme is almost certainly used by the body to inversely permit superoxide to poison bacteria. Superoxide is lethal to cholera.62 Indication that superoxide dismutase is involved in immunity is that phagocytic activity is reduced by free radical scavengers.63

The safest way to transport copper to the immune system would be by the transport protein,59, p.335 ceruloplasmin. This avoids copper toxicity when copper availability to the cells from the liver is increased, since ceruloplasmin copper is not in equilibrium with the serum.59, p.335 The concept that ceruloplasmin is used by the immune cells as a source of copper is supported by the fact that ceruloplasmin quadruples in replete chickens during infection64 and several antigens raise plasma ceruloplasmin in mammals64, p.557 by an unknown hormone, which has been tentatively proposed to be leucocyte endogenous mediator, at low ACTH levels.65, p.557 Cortisol is not used to inversely stimulate ceruloplasmin. I suspect the reason why cortisol is not used is that stress requires extra copper, also, and at high ACTH levels epinephrine is used for this purpose.65, p.556 Transporting copper as the ion is not so important for denying copper to pathogens during digestive disease, which is probably why DOC inversely loses copper from the liver and inhibits liver uptake somewhat thus providing the immune cells with free copper to supplement the ceruloplasmin source.66 Some might argue that it is not likely that the immune cells depend on ceruloplasmin since people with Wilson's disease, in whom ceruloplasmin cannot be synthesized, are not prone to infection. However, such people cannot transport copper to the bile excretory proteins either, so their cells are already loaded and even overloaded with copper.

Cortisol causes an inverse four or five fold decrease of metallothionein,67 a copper storage protein. This may be to furnish more copper for ceruloplasmin synthesis. Cortisol has an opposite effect on alpha aminoisobuteric acid than on the other amino acids.68 If alpha aminoisobuteric acid is used to transport copper through the cell wall, this anomaly would possibly be explained.

MISCELLANEOUS

A large number of other molecules and processes are affected by glucocorticoids which I cannot tie into the immune system definitively at this time. A cursory examination has revealed none to me which is at variance with this thesis. They include smell sensitivity, fear, taste of chloride, pain, appetite, fever, immune cell activity, prostaglandins through arachidonic acid availability, fibronectins, capillary permeability, calcium absorption, intestinal permeability, phosphate, depression, oxidation of chloride, free oxygen formation, blood platelet activating factor, T-cell growth factor sensitivity, and lysosome membrane. Some of these are thought to be controlled by a second message protein, lipocortin, via its effect on phospholipases.69

ENDOTOXIN

Many gram negative bacteria have evolved a very potent way of subverting the cortisol control of immunity. They have a lipopolysacharride called endotoxin on their cell wall. Some endotoxin erodes off the wall and more is released into the blood stream when polymorpholeucocytes eject debris from bacteria which they have engulfed.70 The lipid A part of the molecule stimulates the hypothalamus to secrete large amounts of CRF (corticotrophin releasing factor). An amount of endotoxin which causes no other symptoms than a mild fever causes a six fold rise in ACTH, undoubtedly from CRF stimulation.71 When this way of bypassing ACTH immunity control first arose, it must have been catastrophic for vertebrate life.

A way of detecting endotoxin has apparently evolved and, also, a way of using it to activate a number of responses, some of which are reminiscent of glucocorticoids' inverse effects. Some responses are fever, creation of interferon by spleen cells as well as division of spleen cells, synthesis of IL-6, activation of complement by three mechanisms, creation of hypotension (low blood pressure), stimulation of adherence and oxidative processes of neutrophiles, activation of a burst of activity in macrophages in extremely small amounts, proliferation and maturation of B-cells, suppression of cholera toxin, low serum glucose, metabolic acidosis, and numerous other functions.73 Mice which lack these capabilities are susceptible to gram negative disease.73 Most of these responses are mediated by the peptide hormone cachectin, also called cachexin, or tumor necrosing factor (TNF) secreted by macrophages and they last only the first couple of hours.74 That the detection and cathectin system evolved after the endotoxin assault on ACTH evolved is indicated by the much different appearance of the response curve for endotoxin as opposed to cachectin .99 If both cachectin and gamma interferon are removed by antibodies, bacteria proliferate very rapidly to the host's death. Lipid A fraction of endotoxin enhances local IgA response to mucosally applied antigen [cholera toxin], at least when lipid A and antigen are associated on a liposome carrier.75 GRMFs' secretions are stimulated by endotoxin.76 Antidiuretic hormone quickly rises twenty fold in only 15 minutes.77 Endotoxin must therefore be acting directly on the source of this hormone through cachectin, in my view by a secondary evolutionary response of the hosts. Thus, the body forces endotoxin to mount a preliminary quick response even before the antigens can activate a response, and then quickly turns it off again assisted by a cachectin half life of only six minutes.78

The release of endotoxin by phagocytosis mentioned above is probably the reason why glucocorticoids inhibit digestion but not uptake of bacteria by macrophages.79 This mechanism probably gives the body time to mount its cachectin, GRMF, antibody to endotoxin, and other defenses before the endotoxin containing cell walls are released into the serum.

It would be advantageous if ACTH production could be cut off when under attack. Possibly two proteins detoxify endotoxin.80 Apparently, a mechanism has evolved to cause endotoxin to lose its ability to force ACTH secretion in a few hours.81 This loss may be difficult to control because lymphocytes have developed the ability to secrete a protein, interleukin 1 [IL-1], which has a function of stimulating cortisol secretion5, which it does indirectly by stimulating corticotropin releasing factor (CRF)97, as does IL-6 (the mode of IL-6 action is unknown to me). In other words, the immune system takes over its own regulation. Such a system would be necessary if the ACTH decline were severe because even the immune system cells require maintenance amounts of glucocorticoids. Those glucocorticoid hormones cause the immune cells to rise to a peak of activity at low concentrations and then decline again at increasing concentrations.82 The IL-1 system has an excellent negative feedback.83 IL-1 still retains at least part of the fight or flight override, because it is synergistic with CRF in its long term effects. Cachectin also stimulates ACTH production somewhat by a direct effect on the pituitary,84 possibly an advantage the first few hours, especially if the shutdown of ACTH is rapid.

It would seem desirable if the excess cortisol could be destroyed and, indeed, the half life of cortisol becomes markedly reduced.83 What really makes the IL-1 system practical, however, is the development of a glycoprotein produced by T-cells called glucocorticoid response modifying factor (GRMFs, also GAF) which along with IL-1 has the power to inhibit the response of immune cells to cortisol.6 In other words, the set point of cortisol is raised. Thus, the now multiple sources of ACTH stimulation can be accommodated.

The GRMF system has taken on an advantage not enjoyed by the previous cortisol only control. Since GRMFs do not inhibit cortisol's effect on the immune suppressor cells,6 as previously mentioned, the other immune cells must be stepped up to an even greater frenzy. I suspect a primary pressure forcing the evolution of this system was the advent of endotoxin. The pressure must have been intense because some very virulent diseases are endotoxin involved. They include cholera, typhoid, pneumonia, salmonella, campylobacter, and meningitis. Non-gram negative malaria may also synthesize endotoxin85 perhaps, but if so, probably by some ancient recombinant gene event. Evidence has not been obtained yet that GRMFs affect most of the physiological processes affected by cortisol other than immune cell activity. However GRMF does block phosphoenolpyruvate and fails to block Dibutyryl cyclic AMP induced enzyme synthesis and tyrosine aminotransferase.86, 87 I am not familiar enough with these systems to be able to comment on the significance of these phenomena to the immune system.

CONCLUSIONS

If glucocorticoids are truly immunocorticoids as suggested, it should be possible to use existing information to devise strategies for dealing with infection. It would seem likely that keeping the patient free of stressful thoughts and actions, warm,88 on a low food intake [except for virus], and on a high copper intake (prior to infection) would be advantageous. One way to decrease stress seems to be by way of massage.90 If the patient cannot be guarded from stress, then vitamin C (ascorbic acid) supplements would probably be useful, for they are said to have the effect of blocking a rise in corticosterone resulting from stress91. There is a discussion of diseases for which vitamin C would be advantageous, for some, very advantageous. The advantage may disappear at other times because corticosterone is said to rise some, normally.91 Making sure the patient has ample water during serum disease is probably advantageous because of the effect water status has on corticosterone as mentioned under "Water." Fasting at the noon meal may prove to be a good strategy since cortisol shows a surge then if one eats, but not at the evening meal.92>

Also, 250 watt infrared heat lamps creating an artificial very high fever89, 98, especially on tooth abscesses113, directly on the infected part (except for fungae [personal observation] ), probably are very effective. The immune system is stimulated by a rise in temperature. This could be a response arising by interleuken-1 106. This has been demonstrated for interleuken –1 and interleukin-2 in post operative hypothermia 107. Heat also stimulates tumor necrosis factor (TNF) 108. It could be the reason why the ability to create a fever arose 109. Doubling time of pneumococcal meningitis in rabbits is increased at fever temperature, and that bacteria did not grow at all at 41 degrees centigrade in either soy broth or cerebral fluid 110. So it seems that the efficacy of body temperature effectiveness is dependent on more than enhancement of the immune system. It is conceivable in view of their results that rather than the fever evolving in order to enhance an innate characteristic of the immune system, the fever evolved to take advantage of an innate ineffectiveness of most bacteria at high temperatures and the immune system evolved to be most effective during a fever. It is important to create an artificial fever as soon as possible, because it is most effective early on 111.

The efficacies of these strategies should be established as soon as possible with controlled experiments on primates and made known to the public early on. Such experiments would prove to be very cost effective indeed compared to hospitalization. To rely on hunches based on knowledge of similar chemistry, old wives' tales, and alterations of symptoms by chemicals, such as even the medical profession does currently, is sad and inane. Few will alter their life styles unless they are convinced that the matter is established. It is highly desirable that the theory behind any parameter be understood because even small variations in the patient's environment can sometimes make an otherwise desirable strategy backfire. Nutrition intake and ingestion of poisons and medicines vary wildly in our society, so that treatments based solely on empirical studies such as is the usual case at present in the medical profession can be more than mildly disadvantageous in particular instances. It simply is not possible to take anything for granted in the absence of an experiment. I strongly suspect that the current attitude of the medical profession that potassium can never be deficient, or that rheumatoid arthritis can not possibly be a chronic potassium deficiency even though no experiment has ever been performed, will prove to be tragically wrong, for instance.

In addition, there seems to me to be implied possibilities for clinical intervention against virulent diseases. A recombinantly produced antibody against ACTH or CRF could conceivably have considerable value early in diseases which force their secretion. Perhaps even more valuable and safer would be an antibody against endotoxin. Infection is like a waste paper basket fire. It should be snuffed early before it becomes a raging inferno. Recombinant GRMFs might also prove valuable early in almost any disease. Where GRMFs might prove invaluable at all stages could be in those diseases which compromise the T-cells, such as AIDS, and thus hopefully solve the possible relative excess of glucocorticoids in AIDS.93 Of course, the frequency of injections for peptides must take into account the half life of the peptide to be effective. Massive daily doses would be ineffective and possibly dangerous in many cases. Ceruloplasmin injections would probably be in order for people known to be in a copper deficiency.

It seems conceivable that if a strain of cholera bacteria could be developed which could not synthesize c-AMP toxin or any other toxin, encapsulated in enteric tablets in order to bypass the stomach acids, and swallowed in large amounts, it could act as a preventative to cholera during an epidemic by furnishing overwhelming competition to virulent cholera in the intestines. It might even be effective after an infection.

In any case, it seems to me to be very foolish to administer cortisol to any class of people whose immune system is known to be weak, such as arthritics. If it is desired to raise cortisol's affect in the body, why not use something safe like potassium supplements, or better and safer yet, leafy unboiled vegetables?17,p.447 At the same time, it would solve the problem of the other affects of low whole body potassium content which consistently afflicts arthritics. Arthritics have been shown to improve with a vegetable diet.94 Arthritics have normal cortisol,95 so the lower number of glucocorticosteroid receptors,95 or possibly an abnormal GRMF secretion, must be involved, perhaps triggered by the potassium deficiency itself or some poison. Attempting to solve the problem by injecting cortisol for more than a short time strikes me as dangerous. One author summed it up thus "It is amazing how effective cortisol is in getting a seemingly hopeless patient on his feet again. Sometimes it is so effective that he can walk all the way to the autopsy table". Cortisol is not a medicine, it is a hormone, a hormone whose effects ramify through multiple functions in most of the cell groups in the body. An indication of how fundamental it is, is that the liver's RNA synthesis in adrenalectomized rats is simulated 2-3 fold by cortisol.96 It is urgent that the effects of every known essential nutrient and poison known to be currently ingested be tested against arthritis, especially potassium, which last has never been tested.

The immune system is extremely important to us, so current exploration of immunity should continue on by all known means. However, as you explore, please differentiate between cortisol and corticosterone, use the natural versions, use physiological quantities for at least part of the experiment, use animals other than rodents, and translate jargon. As to this last, immunity is important and extremely complicated. Few theorists are expert in all phases of it and all vocabulary.

Under no circumstances should recombinant experiments be performed which give to any microbe the ability to synthesize cortisol, ACTH, CRF, or any hormone molecule which declines in concentration or effect during infection. No experiment of any kind should be performed on any microbe which synthesizes endotoxin, such as Escheriischia coli. There are thousands of other species.

EPILOGUE

The health of people in the USA is abysmal (numerous statistics), and a major part of it is poor nutrition. As the 12th century physician, trying to cure by diet before he administers drugs, said; “No illness that can be treated by diet should be treated by any other means" or as Hippocrates expressed it in 460 - 377BC; "If we could give every individual the right amount of nourishment and exercise, not too little and not too much, we would have found the safest way to health." It would seem that a healthy life style has been known for a long time. It is my belief that an unprocessed, unfrozen, not canned, high in vegetables diet would keep a large majority of people reasonably healthy and without the need for fad diets. 80% of Americans do not eat adequate vegetables, but even though 72% of Americans take vitamin or mineral supplements daily or sometimes [Sardi p148], their health is atrocious, especially old people..

I would suggest that a partial solution to the problem of poor potassium nutrition would be to place a tax on all food that has had potassium removed by food processors and completely fund all Medicare and workman’s compensation for injuries and disease that relate to rheumatoid arthritis, heart disease, and high blood pressure. This would also take the onerous tax burden now incurred for them and place it on the shoulders of those who cause the problem

The author, Charles Weber, has a degree in chemistry and a masters degree in soil science. He has researched potassium for 45 years, primarily a library research. He has cured his own early onset arthritis (33 years old). He has published articles on allied subjects in; The Journal of Theoretical Biology (1970, 1983), The Journal of Applied Nutrition (1974), Clinical and Experimental Rheumatology (1983), and Medical Hypotheses (1984, 1999).

Joe and Petra:  I am interested in the paper that discusses using hair to measure cortisol levels.  Could you please forward it to me?  Thank you!  

Kristina Babbitt

Hello,

I am IT student and I have to develop a prototype which can measure, process and send salivary cortisol to another hardware. At the moment, the greatest difficulty I am facing is to read the cortisol values from saliva. My question is,"Is there sensor available in the market which can measure the cortisol values????" If not then please guide me some simplest method to measure the cortisol values....

Thanks and looking forward for your response......

Hi Ahsen, I have just sent you an e-mail

I would like to measure Cortisol in tissue (adrenal gland). Does anybody have experience with this matrix? I would need an extraction protocol,..

Many thanks, Barbara Riond

Hi, has anyone injected cortisol into mice aimed at mimicking  the chronic psychological stress? If yes, what was the dose per injection used, 10mg/kg? Thank you!

Zenker, N. & Bernstein, D. E. The estimation of small amounts of corticosterone in rat plasma. J. Biol. Chem. 231, 695-701 (1958).

For chronic stress serum cortisol to salivary cortisol which is better provide adequate counseling can be provided prior to venepuncture.... Does any body have a study on chronic stress bio markers.... 

Different methods can assess different time periods. Are you interested in acute or chronic stress? Overall HPA functioning? This will determine the superior method.

The only reliable saliva measure is to send away to a lab. Hair cortisol is another measure that could be used which measures chronic or historical cortisol over 3 months. It is the HbA1c (referring to glucose off course) of cortisol and hence stress measurements. We are one of few in the world offering this and the only to reliably measure multiple hormones.