Salivary Hormone Testing

Saliva is increasingly being used as a preferential biological fluid for the detection of many different biochemical markers.
Sample collection is non-invasive and much more convenient for both patient and health worker in comparison to, for example, the taking of blood samples, particularly when a number of sequential samples may be required during one 24 hour period. Samples can be collected in the quiet and comfort of the home environment with no additional expertise or help required. Full written instructions are provided.

Adrenal Stress Profile Test

Stress is a major underlying cause of many chronic illnesses, from Chronic Fatigue Syndrome to food and environmental allergies. A stressful lifestyle can lead to consistently high levels of Cortisol in blood and saliva.
The adrenal glands which are found just above the kidney are probably one of the most important glands, with a greater amount of blood passing through them than any other gland in the body. These glands help us deal with stress by releasing a number of hormones, Cortisol and DHEA. Some, including cortisol are released in a cycle called the circadian rhythm, with the highest values being in the morning gradually falling to midnight.
Cortisol elevation also impacts on immune responses, such as secretory IgA (sIgA) and antigliadin antibody (AGA) production.
There are a number of factors which will cause the adrenal glands to become stressed such as long-term mental or physical stress, poor nutrition, smoking, stressful work, eating disorders, divorce, death, intense exercise, alcohol consumption, severe illness or major surgery. In such situations, the hormonal levels become imbalanced. When the Adrenal Gland is functioning normally, we feel well, have adequate amounts of energy and good immunity against infections.

Who Should Have the Test?

The Adrenal Stress Profile test is a measure of an individual’s response to stress. It is also an important tool for pointing to adrenal imbalances that may be impacting a patient’s health.
The Adrenal Stress Test would be particularly valuable for those with a higher risk of adrenal problems:

• People in stressful situations
• Chronic tiredness
• Thyroid disease
• Allergies
• Food intolerance
• Skin problems
• Irritable bowel
• Heart disease
• Blood pressure imbalance

Secretory IgA

Secretory IgA (SIgA) is found throughout the gastrointestinal tract, and in mucus secretions throughout the body and is an important immunoglobulin as a first line of defence against.
Very high levels of sIgA can be found in people who have chronic infections and whose immune system is overloaded or hypersensitive. Lifestyle, stress and nutritional factors can all influence sIgA levels. A deficiency of sIgA is quite common. Low levels of Secretory IgA can make us more susceptible to infection and may be found in many of the following problems:

• Asthma
• Autoimmune conditions
• Coeliac Disease
• Chronic infections
• Crohn's Disease
• Candidiasis
• Food intolerances
• Allergies

Test Requirements:

The Adrenal Stress Profile test gives an assessment of the adrenal hormone cycle by measuring the adrenal rhythm of both salivary cortisol and DHEA-s levels. 
The test requires four saliva samples collected at specific times throughout the day, starting in the morning and finishing at midnight. If a single Cortisol or DHEA-s is required a sample is usually collected at 8am on one day.
The Secretory IgA test requires a sample collected at noon on one day.

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Salivary Sex Hormone

Progesterone

Progesterone is produced in the cortex of the adrenal glands in both sexes as a precursor of the other steroid hormones synthesized at this site. Besides this, a much greater amount of progesterone in females is produced in the cells of the Corpus Luteum in the ovaries and during the pregnancy in the placenta. Small amounts of progesterone are also produced in the testicles of males and also in the brain of both sexes.
The most important role progesterone plays in females is to transform the mucous membrane of the uterus from the proliferation to the secretion phase and therefore to prepare the uterus for the implantation of an eventually fertilized ovum. Progesterone is also responsible for maintaining early pregnancy.

Indications

Regarding the physiological role of progesterone, the main indication for assessing its concentration is the dysregulation in females in puberty, in the premenopausal and the perimenopausal phases. There are some other indications for the measurement of progesterone like control of early pregnancy, evaluation of some ovarian neoplasms etc
There will be no implantation of the ovum in the endometrium of the uterus if the progesterone concentration is too low. It is much more common that the progesterone level will be within the normal range but the shape of the progesterone curve during the cycle is irregular; there may be a shortened luteal phase because of an early degeneration of the corpus luteum. Degeneration may also present with a profile with a premature luteinisation. On the other hand, beginning the luteinisation too late after ovulation may result in an insufficient preparation of the mucous membrane for implantation.
The assessment of salivary progesterone (and oestradiol) is also appropriate to differentiate between conception and non-conception cycles in women. Whereas the oestradiol peak just before ovulation is similar in both kinds of cycles, the progesterone concentration decreases in non-conception cycles (about 7 days following ovulation). In conception cycles a further increase can be observed.
To assess a complete progesterone profile during a cycle, repeated samples have to be taken. This might be inconvenient for patients if blood samples are taken. Therefore saliva collection represents a good alternative.
The following figure summarizes the endocrine situation during the menstrual cycle and its effect on the ovaries and the endometrium of the uterus:

Oestradiol

Oestradiol is the most potent oestrogen of a group of endogenous oestrogen steroids, which also includes oestrone and oestriol. Oestradiol is responsible for maturation of long bones, development of breasts, reproductive organs and secondary female characteristics. Oestradiol is mainly produced by the ovaries with secondary production by the adrenal glands
Oestradiol is a C18 steroid hormone with a phenolic A ring. This steroid hormone has a molecular weight of 272.4. It is the most potent natural oestrogen, produced mainly by the Graffian follicle of the female ovary and the placenta, and in smaller amounts by the adrenals, the male testes and in conversion of steroid precursors into oestrogens in fatty tissue.
Oestradiol is secreted into the blood stream where 98% of it circulates bound to sex hormone binding globulin (SHBG) and to a lesser extent to other serum proteins such as albumin. Only a small fraction circulates as free hormone or in the conjugated form. Oestrogenic activity is effected via oestradiol-receptor complexes which trigger the appropriate response at the nuclear level in the target sites. These sites include the follicles, uterus, breast, vagina, urethra, hypothalamus, pituitary and to a lesser extent the liver and skin.
In non-pregnant women with normal menstrual cycles, oestradiol secretion follows a cyclic, biphasic pattern with the highest concentration found immediately prior to ovulation. The rising oestradiol concentration is understood to exert a positive feedback influence at the level of the pituitary where it influences the secretion of the gonadotrophins, follicle stimulating hormone (FSH), and luteinising hormone (LH), which are essential for follicular maturation and ovulation, respectively. Following ovulation, oestradiol levels fall rapidly until the luteal cells become active resulting in a secondary gentle rise and plateau of oestradiol in the luteal phase. During pregnancy, maternal serum oestradiol levels increase considerably, to well above the pre-­ovulatory peak levels and high levels are sustained throughout pregnancy.
Oestradiol measurements are a valuable index in evaluating a variety of menstrual dysfunctions such as:

• precocious or delayed puberty in girls
• primary and secondary amenorrhea and menopause

Oestradiol levels have also been reported to be increased in patients with feminising syndromes, gynaecomastia and testicular tumours.
In cases of infertility, Oestradiol measurements are useful for monitoring induction of ovulation following treatment with, for example, clomiphene citrate, LH-releasing hormone (LH­RH), or exogenous gonadotrophins. During ovarian hyperstimulation for in vitro fertilization (IVF), oestradiol concentrations are usually monitored daily for optimal timing of human chorionic onadotrophins (hCG) administration and oocyte collection.

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Testosterone

In males almost the whole portion of testosterone is produced in the Leydig cells of the testicles. The release of testosterone is regulated by LH (leuteinising hormone) from the anterior part of the pituitary gland. The FSH (follicle stimulating hormone) from the same site has a positive effect on the Sertoli cells of the testicles which produce the androgen binding protein and therefore activate the development of the sperm cells, which is mediated by the testosterone in the cells. Testosterone itself has a negative feedback on the release of GNRH (gonadotrophin releasing hormone) in the hypothalamic region and FSH and LH in the pituitary gland.
In females the testosterone level in the blood is 10 to 20 times lower than in males. Half of the produced testosterone is derived from the cortex of the adrenal glands and the other half from the ovaries. The release of testosterone from the ovaries is regulated via GNRH of the hypothalamic region and by FSH and LH of the pituitary gland, whereas the production of testosterone from the adrenal cortex is adjusted via the CRH (corticotrophin releasing hormone) from the hypothalamic region and by ACTH (adrenocorticotrophic hormone) from the pituitary gland.
Normally the testosterone concentration in blood and saliva should be higher in the morning in females and males but there is no significant morning peak as with the diurnal cortisol profile. It has to be assumed that the testosterone level in men is fluctuating similar to that of progesterone and oestradiol in women because of the fluctuating release of GNRH and LH in the hypothalamic region and the pituitary gland.
In the blood circulation only 1 - 2 % of the testosterone is not bound to proteins. Only this free portion of testosterone has endocrine effects on the target cells. 35 to 80% of the circulating testosterone is bound to SHBG (sex hormone binding globulin); the remaining part is fixed to albumin. As with other steroid hormones, the levels of the binding globulins and therefore the concentration of bound testosterone is dependent on many physiological and pathological situations in humans. The fraction of free testosterone in blood can be calculated by the values of the whole testosterone concentration and that value of SHBG which implicates the addition of the variability of these two assays. On the other hand the fraction of free testosterone in serum can be directly measured. The quality of these assays is strongly dependent on which amount of the 50 to 100 times higher protein bound testosterone portion is also measured by the test.
In the salivary glands the free testosterone is wholly separated from the protein bound fraction. The salivary testosterone level, therefore, actually reflects the level of the active part of the hormone.

Test Requirements:

The Salivary Female Hormone Profile test requires eleven saliva samples collected around noon on specific days throughout the monthly cycle. (Because hormone release is episodic, it is preferable to collect the sample over two hours; i.e. 11am to 1pm.)
The Salivary Sex Hormone Profile and Salivary Male Hormone Profile requires four saliva samples collected a specific times throughout the day, starting in the morning and finishing at midnight.
The Salivary Sex Hormones only requires a sample collected around noon on one day. (Because hormone release is episodic, it is preferable to collect the sample over two hours; i.e. between 11am and 1pm.)

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Melatonin

Melatonin is a hormone produced by the pineal gland. It secretes melatonin in a circadian fashion. It has an in built rhythm of about 24.5-25.5 hours. Levels start to rise at around 8.00 p.m. and are maximal at 2-3.00 a.m. and fall by 8.00 a.m. It is the main internal timegiver to the body and is the key neuroendocrine modulator of annual and Circadian biorythms in man. The intrinsic rhythm can be altered by external clues such as the light/dark cycle, social clues such as an alarm clock going off or having to go to work and also therapeutically by light therapies and can be interfered with by various pharmaceutical agents. The commonest are beta blockers which block the beta receptors of the pineal gland inhibiting melatonin synthesis producing insomnia and non-steroidal anti-inflammatories which can inhibit melatonin synthesis also.
Rhythms are important to the body as they control chaos. Integration of the endocrine, immune and nervous systems are very important for communication, co­ordination and control over the body's processes. Rhythmic changes in hormone levels are the main way stability is ensured in the body. Clinically, problems can arise with various situations. The commonest encountered are those concerning the effects of jet lag and shift work. However, melatonin abnormalities have been demonstrated in the so called seasonal affective disorder which is where people have high levels of winter time melatonin accompanied by carbohydrate craving. More recently abnormalities have been found in people suffering from CFS (ME) and may be one of the reasons their illness is prolonged. Biologically its actions are:
1.         Sleep induction via the histamine interleukin prostaglandin pathway.
2.         Buffering of stress hormones such as cortisol.
3.         Immune system conditioning.
4.         Anti-oxidant effects, being the body's most powerful natural anti-oxidant.
5.         Potentiation of other hormones, e.g. T4 to T3 conversion is enhanced by melatonin.
6.         Inhibition of gonadotrophin hormones.
There is an interesting group of patients who often present with multiple food allergies/sensitivities. A common symptom patients have is feeling bloated after food.
They can be shown on occasions to have high levels of melatonin released from the gastrointestinal tract after eating — this can be treated with anti-histamines. All treatments should last between 4-6 months. Hopefully, the external imposition of a rhythm will entrain the brain to produce a normal rhythm of its own after this time.
In comparison to serum levels, salivary melatonin concentrations in the night time are generally lower. However the position of the melatonin profile (acrophase) does not differ within a subject in the concomitant measurement of serum and salivary melatonin. Therefore, the measurement of salivary melatonin levels seems valid for studies on melatonin rhythms (1)
(1)        Maurizi C P : The therapeutical potential for tryptophan and melatonin: Possible roles in depression, sleep, Alzheimers disease and abnormal aging Medical Hyp1990)

Test Requirements:

The Salivary Sleep Profile & Melatonin Profile requires four saliva samples collected at specific times throughout the day, starting in the morning and finishing at midnight.
The Salivary Single Melatonin only requires a single sample collected at midnight or as required.

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