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Canary ClubAdrenal Articles

Clin Endocrinol (Oxf). 2009 Apr;70(4):516-21. Epub 2008 Aug 15.

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Diagnostic value of salivary cortisol in Cushing's syndrome (CS).

Cardoso EM, Arregger AL, Tumilasci OR, Contreras LN.

Endocrine Research Department, Instituto de Investigaciones Médicas A. Lanari, School of Medicine, University of Buenos Aires, Argentina.

Abstract

OBJECTIVE: The diagnosis of Cushing's syndrome (CS) remains a challenge in clinical endocrinology. The aim of this study was to determine the reproducibility and diagnostic value of late-night salivary cortisol (SAF(23)) for CS and its utility along the follow-up of treated patients. In addition, using the same radioimmunoassay reactives, the cut-off values for saliva and serum cortisol, assessed synchronically after the overnight 1 mg dexamethasone suppression test (DST), were defined.

DESIGN: Twenty-one patients with confirmed CS and 121 volunteers were studied. All the subjects collected 24-h urine for cortisol (UFC). On the same day whole saliva was obtained from the subjects at 23 h for SAF(23). The intraclass coefficient of correlation (ICC) of SAF(23) was estimated in 47 subjects (21 CS and 26 C). At 8 h, after DST, simultaneous saliva and serum samples for cortisol (SAF(dex) and F(dex), respectively) were obtained in 51 subjects (17 CS and 34 C). After specific therapy, 18 patients with CS were followed with SAF(23) measurements. SAF and F were expressed as nM.

RESULTS: The intraclass coefficient of correlation of SAF(23) was 0.89 in CS and 0.83 in C. SAF(23) > 3.8 nM showed a sensitivity and specificity of 100% and 97.5%, respectively, for diagnosing CS. SAF(23) correlated positively with UFC (r = 0.685; P = 0.0001). After DST, SAF(dex) significantly correlated with F(dex) (r = 0.61, P < 0.0001). A cut-off value of SAF(dex) > 2.0 nM and F(dex) > 50.0 nM detected CS with 100% sensitivity and specificity. After successful surgical therapy, 13 patients with CS had SAF(23) levels < 3.8 nM (1.4 +/- 0.8 nM).

CONCLUSIONS: SAF(23) and SAF(dex) seem to be good screening tools based on their noninvasive nature, remarkable reproducibility and diagnostic performances. 
 

Clin Chem. 2008 Nov;54(11):1759-69. Epub 2008 Aug 29.

Current status of salivary hormone analysis.

Gröschl M.

Department of Pediatrics, University of Erlangen-Nürnberg, Erlangen, Germany. michael.groeschl@uk-erlangen.de

Abstract

BACKGROUND: Saliva, which offers a noninvasive and stress-free alternative to plasma and serum, is a widely accepted sample source for analysis of steroids and also of certain amines and peptides. In recent years, numerous publications have described the use of salivary hormone analysis in many fields of clinical and basic research.

CONTENT: This review provides an overview of the current applications of salivary hormone analysis. A description of the different modes of hormone entry into saliva is followed by a detailed description of analytical methods and approaches for reliable collection of saliva, including several interesting applications in diverse fields including psychiatry, stress research, clinical endocrinology, sports medicine, and veterinary medicine.

SUMMARY: Although saliva has not yet become a mainstream sample source for hormone analysis, it has proven to be reliable and, in some cases, even superior to other body fluids. Nevertheless much effort will be required for this approach to receive acceptance over the long term, especially by clinicians. Such effort includes the development of specific and standardized analytical tools, the establishment of defined reference intervals, and implementation of round-robin trials. One major problem, the lack of compliance sometimes seen in outpatient saliva donors, requires strict standardization of both collection and analysis methods to achieve better comparability and assessment of published salivary hormone data.

PMID: 18757583 [PubMed - indexed for MEDLINE]Free Article 
 
 
 

Clin Endocrinol (Oxf). 2007 Nov;67(5):656-62.

Salivary testosterone: a reliable approach to the diagnosis of male hypogonadism.

Arregger AL, Contreras LN, Tumilasci OR, Aquilano DR, Cardoso EM.

Endocrine Research Department, Instituto de Investigaciones Médicas A. Lanari, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina.

Abstract

OBJECTIVE: This study was to demonstrate that Sal-T is a reliable biomarker of androgen status in the diagnosis of male hypogonadism.

DESIGN: In order to validate the salivary testosterone assay (Sal-T), its reproducibility, the agreement with serum free testosterone levels (Free-T), the correlation with other circulating androgen markers (bioavailable testosterone, total testosterone) and cut-off values were defined.

PATIENTS AND METHODS: We studied 52 eugonadic (E) and 20 hypogonadic (Hy) men. Sal-T was assayed using an adapted radioimmunoassay for serum testosterone. Sal-T concentrations were compared in nine cases before and after citric acid stimulation of salivary flow rate. Free-T and bioavailable testosterone (Bio-T) were calculated by Vermeulen equation and SHBG were determined by binding assay.

RESULTS: Sal-T did not depend on salivary flow rate and morning samples from 07.00 h to 09.00 h were stable. Agreement between Sal-T and Free-T measurements was confirmed in all subjects. Sal-T levels correlated positively with all circulating androgens, showing the best correlation with Free-T in E (r = 0.92) as well as in Hy (r = 0.97). A cut-off value of Sal-T < or = 0.195 nm showed 100% sensibility and specificity to rule out hypogonadism.

CONCLUSIONS: Our data showed that Sal-T is a reliable marker of testosterone bioavailability. The results support the inclusion of this biomarker as a noninvasive approach in the diagnosis of male androgen deficiency.

PMID: 17953627 [PubMed - indexed for MEDLINE] 
 
 

Clin Endocrinol (Oxf). 2005 Sep;63(3):336-41.

Salivary cortisol determined by enzyme immunoassay is preferable to serum total cortisol for assessment of dynamic hypothalamic--pituitary--adrenal axis activity.

Gozansky WS, Lynn JS, Laudenslager ML, Kohrt WM.

Division of Geriatric Medicine, Department of Medicine, University of Colorado at Denver and Health Sciences Center, Denver, CO 80262, USA. wendee.gozansky@uchsc.edu

Abstract

OBJECTIVE: The aim of this study was to determine whether salivary cortisol measured by a simple enzyme immunoassay (EIA) could be used as a surrogate for serum total cortisol in response to rapid changes and across a wide range of concentrations. DESIGN: Comparisons of matched salivary and serum samples in response to dynamic hypothalamic-pituitary-adrenal (HPA) axis testing. Subjects Healthy women (n=10; three taking oral oestrogens) and men (n=2), aged 23--65 years, were recruited from the community. Measurements Paired saliva and serum samples were obtained during three protocols: 10 min of exercise at 90% of maximal heart rate (n=8), intravenous administration of corticotrophin-releasing hormone (CRH; n=4), and dexamethasone suppression (n=7). Cortisol was measured in saliva using a commercial high-sensitivity EIA and total cortisol was measured in serum with a commercial radioimmunoassay (RIA). Results The time course of the salivary cortisol response to both the exercise and CRH tests paralleled that of total serum cortisol. Salivary cortisol demonstrated a significantly greater relative increase in response to the exercise and CRH stimuli (697+/- 826%vs. 209+/- 150%, P=0.04 saliva vs. serum). A disproportionately larger increase in free cortisol, compared with total, would be expected when the binding capacity of cortisol-binding globulin (CBG) is exceeded. In response to dexamethasone suppression, relative decreases in cortisol were not significantly different between the two media (-47+/- 56%vs.-84+/- 8%, P=0.13 saliva vs. serum). Although a significant linear correlation was found for all paired salivary and serum total cortisol samples (n=183 pairs, r=0.60, P<0.001), an exponential model provided a better fit (r=0.81, P<0.001). The linear correlations were strengthened when data from subjects on oral oestrogens (n=52 pairs, r=0.75, P < 0.001) were separated from those not taking oestrogens (n=131 pairs, r=0.67, P<0.001). Conclusions Salivary cortisol measured with a simple EIA can be used in place of serum total cortisol in physiological research protocols. Evidence that salivary measures represent the biologically active, free fraction of cortisol includes: (1) the greater relative increase in salivary cortisol in response to tests that raise the absolute cortisol concentration above the saturation point of CBG; (2) the strong exponential relationship between cortisol assessed in the two media; and (3) the improved linear correlations when subjects known to have increased CBG were analysed separately. Thus, an advantage of measuring salivary cortisol rather than total serum cortisol is that it eliminates the need to account for within-subject changes or between-subject differences in CBG.

PMID: 16117823 [PubMed - indexed for MEDLINE] 
 
 

Gynecol Obstet Invest. 2002;53(1):32-7.

The clinical usefulness of salivary progesterone measurement for the evaluation of the corpus luteum function.

Ishikawa M, Sengoku K, Tamate K, Takaoka Y, Kane M, Fottrell PF.

Department of Obstetrics and Gynecology, Asahikawa Medical College, Asahikawa, Japan.

Abstract

The present study was designed to construct reliable daily salivary progesterone profiles throughout the luteal phase to accurately evaluate the corpus luteum function. Furthermore, we investigated the clinical relevance of a simple midluteal salivary progesterone estimation for the diagnosis of luteal phase insufficiency by determining the diagnostic efficiency and cutoff values. A total of 121 women were divided into 3 groups; normal luteal function, luteal phase insufficiency and unclassified group, based on basal body temperature recordings and serum progesterone levels at 3 sampling points during the midluteal phase. Salivary progesterone values across the luteal phase of the normal luteal function group were significantly increased from day 1 to day 4, remained constant from day 5 to day 9 (mean +/- SD, 318 +/- 170 pmol/l on day 5, 287 +/- 169 pmol/l on day 9; urinary LH surge = day 0) and decreased thereafter. Salivary progesterone concentrations in the luteal phase insufficiency group showed significantly lower values compared with those in the normal group between days 3 and 10. The cutoff values of 189 pmol/l in the midluteal phase yielded a sensitivity of 78.0% and a specificity of 76.5%. Our results suggest that daily salivary progesterone profiles during the luteal phase and a simple estimation of midluteal salivary progesterone appeared to be useful for the diagnosis of luteal phase defects.

Copyright 2002 S. Karger AG, Basel

PMID: 11803226 [PubMed - indexed for MEDLINE] 
 
 

Cancer Epidemiol Biomarkers Prev. 2001 Jan;10(1):59-64.

Saliva as a medium for investigating intra- and interindividual differences in sex hormone levels in premenopausal women.

Gann PH, Giovanazzi S, Van Horn L, Branning A, Chatterton RT Jr.

Department of Preventive Medicine, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611, USA. pgann@northwestern.edu

Abstract

Repeated measurement of ovarian steroids in saliva could provide an advantage in studies estimating long-term sex steroid exposure in premenopausal women, by reducing the measurement error associated with collection of serum or urine samples. We previously reported on characteristics of ultrasensitive RIAs adapted for extraction-free measurement of estradiol (E2) and progesterone (PG) in saliva. The purpose of the present study was to evaluate the consistency of E2 and PG levels in saliva in the same women across menstrual cycles, and to compare this with the variation observed between women. We also evaluated the effect of altering the number of consecutive daily samples considered and the method for locating a particular cycle day in relation to ovulation (day 0). Study participants included 12 healthy women who provided daily saliva samples for two consecutive, ovulatory menstrual cycles. A single midluteal serum sample was collected 7-8 days after detection of a luteinizing hormone (LH) peak in urine. We plotted individual cycle profiles and computed intraclass correlation coefficients (ICC) for various definitions of peak and cumulative daily hormone level. For peak PG, determined as the maximal running 3-day mean, ICC was 0.68. For cumulative PG, based on 8 consecutive cycle days (+2 to +9), ICCs were 0.72-0.76 when reverse dating LH peak or rise in salivary PG determined day 0. For E2, ICCs ranged from 0.74 to 0.79 by various dating methods for the 5 preovulatory days (-4-0), and from 0.85 to 0.92 for the 15 days about the center of the cycle (-6 to +8). With exclusion of just the first 5 days of the cycle, the ICC for E2 was 0.91. For both E2 and PG, selection of 5 or 7 days for the estimation of the midluteal mean level provided separation of within and between subject variance that was comparable with a LH-timed serum sample. These results indicate that daily saliva samples can be combined to clarify the interindividual differences in E2 and PG levels in premenopausal women, and that these interindividual differences may be greater than previously imagined.

PMID: 11205490 [PubMed - indexed for MEDLINE]Free Article 
 
 

Gynecol Endocrinol. 1993 Jun;7(2):101-10.

Assessment of bioavailability of oral micronized progesterone using a salivary progesterone enzymeimmunoassay.

Bolaji II, Tallon DF, O'Dwyer E, Fottrell PF.

Department of Obstetrics and Gynaecology, University College, Galway, Ireland.

Abstract

Salivary progesterone was measured sequentially by enzymeimmunoassay following 1 month and 6 months of oral therapy with 100 mg of micronized progesterone (MOP) in 40 healthy estrogenized postmenopausal women (aged 40-68 years). MOP was administered for 23 days every month. There were striking differences in the absorption of MOP between various subjects. Significant increases occurred in salivary progesterone concentrations over baseline and pretreatment levels and persisted for at least 10 h. Levels of salivary progesterone remained higher than pretreatment levels for at least 24 h after administration of MOP. Maximum mean concentrations of salivary progesterone of 827.2 and 888 pmol/l in the 1st and 6th months of therapy, respectively, were achieved within 2 h of administration and were above the 95th percentile of a control corridor which corresponds to the range found in the luteal phase. The areas under the salivary progesterone curve (AUC0-24 h, pmol/l) were 7177.75 and 7388.20 respectively, in the 1st and 6th months of therapy but the difference was not statistically significant. Serum and salivary progesterone peaked simultaneously and there was a significant correlation between the concentrations measured concurrently (y = 233.08 + 35.575x; r = 0.89, p < 0.001) thus supporting the current concept of a relatively rapid diffusion of steroids from plasma to saliva. Results of this study confirm those of previous investigations which monitored the bioavailability of MOP with the use of serum progesterone measurements and showed that luteal phase progesterone concentrations can be attained easily. The use of non-invasive salivary sampling and a cost-effective, direct enzymeimmunoassay showed a considerable advantage in the present study, compared with previous ones. We conclude that 100 mg MOP should be given at least twice-daily to maintain a stable physiological luteal phase level of progesterone during clinical hormone replacement therapy.

PMID: 8213223 [PubMed - indexed for MEDLINE] 
 
 

Gynecol Endocrinol. 1993 Jun;7(2):101-10.

Assessment of bioavailability of oral micronized progesterone using a salivary progesterone enzymeimmunoassay.

Bolaji II, Tallon DF, O'Dwyer E, Fottrell PF.

Department of Obstetrics and Gynaecology, University College, Galway, Ireland.

Abstract

Salivary progesterone was measured sequentially by enzymeimmunoassay following 1 month and 6 months of oral therapy with 100 mg of micronized progesterone (MOP) in 40 healthy estrogenized postmenopausal women (aged 40-68 years). MOP was administered for 23 days every month. There were striking differences in the absorption of MOP between various subjects. Significant increases occurred in salivary progesterone concentrations over baseline and pretreatment levels and persisted for at least 10 h. Levels of salivary progesterone remained higher than pretreatment levels for at least 24 h after administration of MOP. Maximum mean concentrations of salivary progesterone of 827.2 and 888 pmol/l in the 1st and 6th months of therapy, respectively, were achieved within 2 h of administration and were above the 95th percentile of a control corridor which corresponds to the range found in the luteal phase. The areas under the salivary progesterone curve (AUC0-24 h, pmol/l) were 7177.75 and 7388.20 respectively, in the 1st and 6th months of therapy but the difference was not statistically significant. Serum and salivary progesterone peaked simultaneously and there was a significant correlation between the concentrations measured concurrently (y = 233.08 + 35.575x; r = 0.89, p < 0.001) thus supporting the current concept of a relatively rapid diffusion of steroids from plasma to saliva. Results of this study confirm those of previous investigations which monitored the bioavailability of MOP with the use of serum progesterone measurements and showed that luteal phase progesterone concentrations can be attained easily. The use of non-invasive salivary sampling and a cost-effective, direct enzymeimmunoassay showed a considerable advantage in the present study, compared with previous ones. We conclude that 100 mg MOP should be given at least twice-daily to maintain a stable physiological luteal phase level of progesterone during clinical hormone replacement therapy.

PMID: 8213223 [PubMed - indexed for MEDLINE] 
 

Gynecol Endocrinol. 1993 Jun;7(2):101-10.

Assessment of bioavailability of oral micronized progesterone using a salivary progesterone enzymeimmunoassay.

Bolaji II, Tallon DF, O'Dwyer E, Fottrell PF.

Department of Obstetrics and Gynaecology, University College, Galway, Ireland.

Abstract

Salivary progesterone was measured sequentially by enzymeimmunoassay following 1 month and 6 months of oral therapy with 100 mg of micronized progesterone (MOP) in 40 healthy estrogenized postmenopausal women (aged 40-68 years). MOP was administered for 23 days every month. There were striking differences in the absorption of MOP between various subjects. Significant increases occurred in salivary progesterone concentrations over baseline and pretreatment levels and persisted for at least 10 h. Levels of salivary progesterone remained higher than pretreatment levels for at least 24 h after administration of MOP. Maximum mean concentrations of salivary progesterone of 827.2 and 888 pmol/l in the 1st and 6th months of therapy, respectively, were achieved within 2 h of administration and were above the 95th percentile of a control corridor which corresponds to the range found in the luteal phase. The areas under the salivary progesterone curve (AUC0-24 h, pmol/l) were 7177.75 and 7388.20 respectively, in the 1st and 6th months of therapy but the difference was not statistically significant. Serum and salivary progesterone peaked simultaneously and there was a significant correlation between the concentrations measured concurrently (y = 233.08 + 35.575x; r = 0.89, p < 0.001) thus supporting the current concept of a relatively rapid diffusion of steroids from plasma to saliva. Results of this study confirm those of previous investigations which monitored the bioavailability of MOP with the use of serum progesterone measurements and showed that luteal phase progesterone concentrations can be attained easily. The use of non-invasive salivary sampling and a cost-effective, direct enzymeimmunoassay showed a considerable advantage in the present study, compared with previous ones. We conclude that 100 mg MOP should be given at least twice-daily to maintain a stable physiological luteal phase level of progesterone during clinical hormone replacement therapy.

PMID: 8213223 [PubMed - indexed for MEDLINE] 
 
 

Clin Chem. 1990 Oct;36(10):1769-73.

Sensitive salivary estradiol assay for monitoring ovarian function.

Worthman CM, Stallings JF, Hofman LF.

Department of Anthropology, Emory University, Atlanta, GA 30322.

Abstract

Measurement of steroids in saliva has excited interest because of the numerous potential clinical applications; noninvasive, convenient sampling; and apparently accurate reflection of the concentrations of physiologically active unbound steroid in the circulation. Although assays of saliva for several steroid hormones are available and widely used, assays for salivary estradiol are not, primarily because of methodological limitations. By modifying a commercially available kit for serum estradiol, our laboratory has developed a procedure that is sensitive, highly specific, and reliable for measuring salivary estradiol. Assay sensitivity is 0.5 fmol (0.14 pg; sample concentration 1.3 pmol/L) with a mean interassay CV of 10.8% at low concentrations. Clinical studies showed that values for serum and saliva are highly correlated (P less than 0.001), and demonstrated reliable detection of estradiol peaks during normal ovulatory cycles in serial samples from 15 women. Salivary estradiol peaked at 5.4 (SD 1.9) pmol/L on cycle day 14.4 (SD 3.2), 1.2 (SD 0.8) days before ovulation detected by ultrasound. This assay may be particularly helpful in investigating ovarian function and free estradiol in women at various stages of the reproductive cycle.

PMID: 2208652 [PubMed - indexed for MEDLINE] 
 
 

Crit Rev Clin Lab Sci. 1986;23(2):95-146.

Hormones in saliva.

Vining RF, McGinley RA.

Abstract

Since the collection of saliva is noninvasive, nonstressful and usually very convenient there have been many recent studies examining the clinical relevance of measuring various hormones in saliva. It now appears that the measurement of most unconjugated steroids in saliva will provide clinically useful data whereas the measurement of conjugated steroids, thyroid hormones, and protein hormones is unlikely to be clinically relevant. The key factors determining whether the salivary concentration of a hormone or drug is likely to be clinically relevant are the mechanisms by which the material enters the saliva; the "free to protein bound" ratio for the material; and the structure of the material, i.e., its molecular weight, polarity and the presence of ionizable groups.

PMID: 3512171 [PubMed - indexed for MEDLINE] 

Publication Types, MeSH Terms, Substances 
 
 

Horm Behav. 2004 Jun;46(1):39-46.

Quantifying blood leakage into the oral mucosa and its effects on the measurement of cortisol, dehydroepiandrosterone, and testosterone in saliva.

Kivlighan KT, Granger DA, Schwartz EB, Nelson V, Curran M, Shirtcliff EA.

Behavioral Endocrinology Laboratory, Department of Biobehavioral Health, The Pennsylvania State University, University Park, PA 16802, USA.

Abstract

The impact of blood leakage due to microinjury to the oral cavity on the measurement of salivary hormones was examined. Saliva samples were collected before, immediately after, and then every 15 min for 1 h following vigorous tooth brushing. Blood in saliva was quantified by visual inspection of discoloration, Hemastix reagent strips to detect hemoglobin, and an immunoassay for transferrin. The presence of blood in saliva immediately after microinjury was confirmed by all methods. Hemoglobin and transferrin levels remained elevated over baseline for at least 30 min. Levels of salivary testosterone increased over baseline and remained elevated for 30 min in response to microinjury. Microinjury induced change in salivary testosterone was more closely associated with the change in transferrin than hemoglobin levels or discoloration ratings. On average, levels of salivary dehydroepiandrosterone (DHEA) did not increase in response to microinjury. However, individual differences in microinjury induced change in DHEA were associated with discoloration ratings. Salivary cortisol levels, on average, were neither responsive to microinjury, nor were individual differences in cortisol change associated with blood contamination measures. Neither diurnal nor gender-related differences in baseline hormone levels predicted the impact of blood leakage on quantitative salivary measurements. The findings suggest ecologically valid minor-to-moderate level microinjuries to the oral cavity have negligible effects on the measurement of salivary cortisol, but may be important to quantify and control when assessing other hormones especially testosterone.

PMID: 15215040 [PubMed - indexed for MEDLINE] 
 
 

J Clin Endocrinol Metab. 2008 May;93(5):1526-40. Epub 2008 Mar 11.

The diagnosis of Cushing's syndrome: an Endocrine Society Clinical Practice Guideline.

Nieman LK, Biller BM, Findling JW, Newell-Price J, Savage MO, Stewart PM, Montori VM.

Program on Reproductive and Adult Endocrinology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. govt-prof@endo.society.org

Abstract

OBJECTIVE: The objective of the study was to develop clinical practice guidelines for the diagnosis of Cushing's syndrome.

PARTICIPANTS: The Task Force included a chair, selected by the Clinical Guidelines Subcommittee (CGS) of The Endocrine Society, five additional experts, a methodologist, and a medical writer. The Task Force received no corporate funding or remuneration.

CONSENSUS PROCESS: Consensus was guided by systematic reviews of evidence and discussions. The guidelines were reviewed and approved sequentially by The Endocrine Society's CGS and Clinical Affairs Core Committee, members responding to a web posting, and The Endocrine Society Council. At each stage the Task Force incorporated needed changes in response to written comments.

CONCLUSIONS: After excluding exogenous glucocorticoid use, we recommend testing for Cushing's syndrome in patients with multiple and progressive features compatible with the syndrome, particularly those with a high discriminatory value, and patients with adrenal incidentaloma. We recommend initial use of one test with high diagnostic accuracy (urine cortisol, late night salivary cortisol, 1 mg overnight or 2 mg 48-h dexamethasone suppression test). We recommend that patients with an abnormal result see an endocrinologist and undergo a second test, either one of the above or, in some cases, a serum midnight cortisol or dexamethasone-CRH test. Patients with concordant abnormal results should undergo testing for the cause of Cushing's syndrome. Patients with concordant normal results should not undergo further evaluation. We recommend additional testing in patients with discordant results, normal responses suspected of cyclic hypercortisolism, or initially normal responses who accumulate additional features over time.

PMID: 18334580 [PubMed - indexed for MEDLINE] 

Scientific Books and Articles

Diagnosis and Treatment of Adrenal Fatigue (mild adrenal insufficiency.)

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