Hanna-Leena Kaihola

Reference intervals for serum sex steroids and gonadotropins in regularly menstruating women

Reference intervals for novel fluoro‐immunoassays for serum luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol (E2) and progesterone (P) were established in 40 healthy regularly menstruating women. Serum samples for sex steroids, including testosterone (T), androstenedione (A) and dehydroepiandrosterone sulphate (DHEAS), as well as sex hormone binding globulin (SHBG) and gonadotropins, were collected in the early follicular and mid‐luteal phases of the menstrual cycle. The need for the timing of androgen and gonadotropin samples was also determinated. Serum E2 and P were measured by radio‐immuno‐ and time‐resolved fluoro‐immunoassays. Various assay methods correlated closely, but the reference intervals varied considerably from one method to another. In the early follicular phase the LH/FSH ratio only ranged between 0.2 and 1.7 (mean ± S.D.; 0.8 ± 0.3); the variation was greater (from 0.3 to 3.5) in the luteal phase, when the mean was also significantly higher (1.5 ± 0.9, p = 0.0001). Similarly the mean values of T and SHBG were higher in the luteal phase, while the other androgen concentrations and calculated androgen indexes were equal, compared with the follicular phase values. Thus the timing of hormone determinations is warranted in the investigation of the gonadotropin and androgen status in normally menstruating women. Moreover, the upper limit of the LH/FSH ratio examined with new, sensitive methods is lower than that previously stated. The use of an intra‐uterine contraceptive device (IUD) had no effect on hormone levels.

C-peptide determination in the choice of treatment in diabetes mellitus

The predictive value of the intravenous glucagon test in assessing the requirement of insulin therapy in diabetes mellitus was evaluated in 105 adult diabetics. Basal and stimulated C-peptide concentrations and increments of C-peptide concentration were examined separately among newly and previously diagnosed diabetics. The poststimulatory C-peptide concentration of 0.6 nmol/l (Novo, antibody M 1230) proved to be the most reliable basis for the choice of therapy. Adequate therapy could have been assessed in 70 cases (67%) without glucagon stimulation. To derive maximal information of plasma C-peptide concentrations, a biphasic scheme of the use for C-peptide determinations and glucagon stimulation is presented. Basal and stimulated C-peptide levels of insulin-requiring diabetics correlated negatively with the duration of diabetes but they did not correlate with the relative body weights. Basal and stimulated C-peptide levels of non-insulin-requiring diabetics did not correlate with the duration of diabetes, but they correlated positively with the relative body weights.

Serum androgen and gonadotropin levels decline after progestogen-induced withdrawal bleeding in oligomenorrheic women with or without polycystic ovaries *

OBJECTIVE To examine the effect of short-term progestogen treatment on androgen, gonadotropin, and sex hormone-binding globulin (SHBG) levels in oligomenorrheic women. DESIGN Comparative study of changes in hormonal parameters in patients with or without ultrasonographically diagnosed polycystic ovarian disease (PCOD). SETTING Open patient clinic of reproductive endocrinology at University Central Hospital of Turku, Finland. PATIENTS Seventy-five oligomenorrheic women with (n = 51) or without (n = 24) PCOD. MAIN OUTCOME MEASURES Serum concentrations of testosterone (T), androstenedione (A), dehydroepiandrosterone sulfate, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and SHBG. RESULTS The levels of T, A, LH, and the LH:FSH ratios decreased significantly after oral treatment with medroxyprogesterone acetate (10 mg/d for 10 days) in non-PCOD women and in women with PCOD decreasing the frequencies of pathological laboratory findings, in particular elevated levels of LH:FSH ratio and A in PCOD women and of LH:FSH ratio in non-PCOD women. The levels of T, A, and LH as well as the LH:FSH ratio were significantly higher in women with PCOD. Obesity was associated with high free androgen indices, low LH:FSH ratios, and low concentrations of LH, A, and SHBG. CONCLUSIONS The serum samples for hormonal analyses used as an aid in diagnosing PCOD should be obtained without pretreatment with progestogen because it masks the biochemical findings of PCOD.

Uterine Size Measured by Ultrasound During the Menstrual Cycle

In the following study changes in the size of non-pregnant uterus were measured by B-scan ultrasonography. Uterine size in different stages of the menstrual cycle was measured ultrasonically in 16 women, whose periods were confirmed to be ovulatory, both by basal body temperature (BBT) and by the radioimmunological measurement of plasma estradiol and progesterone. It was established that the size of the uterus grows significantly towards the end of the menstrual cycle. In addition to the above mentioned, the examination was performed on a woman of child-bearing age with disturbances in her menstrual cycle. Hormone measurements indicated that her period was anovulatory. No typical uterine growth characteristic of ovulatory cycles could be observed.

Plasma and urinary C-peptide in the classification of adult diabetics

Plasma and urinary C-peptide determinations in the discrimination between insulin-requiring and non-insulin-requiring diabetes were elevated in 61 adult diabetics. Specimens for C-peptide determinations were taken on two consecutive days: on the first day plasma C-peptide concentrations were determined before and 6 min after intravenous glucagon administration. On the second day 2- and 4-h urinary C-peptide excretion was measured after an individual breakfast. Results of urinary C-peptide analyses were expressed as molar concentration and also as molar quantity excreted (without any corrections and related to creatinine excretion). Glucagon-stimulated plasma C-peptide turned out to be a reliable criterion for the detection of insulin requirement. Sixty-nine per cent of diabetics included in this study were classifiable by basal plasma C-peptide concentrations. Two-hour postprandial urinary C-peptide/creatinine quotient turned out to be slightly less sensitive (89%) than the glucagon test (94%) and of equal specificity (96%). Glucagon-stimulated plasma C-peptide and postprandial urinary C-peptide excretion correlated significantly among insulin-requiring diabetics (r = 0.73), but not among non-insulin-requiring diabetics (r = 0.23). We regard determination of stimulated plasma C-peptide as a primary investigation for the direct assessment of endogenous insulin secretory reserves for clinical management decisions. Determination of postprandial urinary C-peptide is applicable in selected situations for non-invasive assessment of insulin secretion.

Haematological laboratory findings in the elderly: influence of age and sex

The effects of age and sex on haematological laboratory parameters were studied in connection with a population study in people over the age of 65 years (n = 347). Serum vitamin B12 was the only parameter which decreased significantly with advancing age. Blood leucocyte count, haemoglobin concentration, haematocrit, erythrocyte count, mean erythrocyte volume, mean erythrocyte haemoglobin and serum ferritin values were significantly higher in males than in females. Serum iron, serum transferrin, and plasma and erythrocyte folate levels did not differ between males and females. Thirteen subjects were anaemic and three of them had iron deficiency anaemia. Five subjects had iron deficiency based on serum iron and transferrin but no anaemia. Serum ferritin measurement did not reveal any further subjects with iron deficiency. No case of folate deficiency anaemia was revealed. Although many of the participants were on medication, most of them were living at home and taking care of themselves and represent relatively fit elderly people. Therefore we suggest that these laboratory data can also serve as reference values for the elderly people.

The grade of hirsutism correlated to serum androgen levels and hormonal indices

Abstract. The degree of hair growth of 64 hirsute women was clinically graded. The patients were divided into two groups with respect to the ratio between the serum luteinizing hormone (LH) and the follicle‐stimulating hormone (FSH). The two groups did not differ in terms of the grade of hirsutism, serum testosterone (T), calculated free testosterone (FTc), androstenedione (A), dehydroepiandrosterone sulphate (DHEAS), sex hormone binding globulin (SHBG), estradiol, or cortisol levels. In patients with a LH/FSH ratio of 3.0 or less (n = 49), FTc and A correlated well (ϱ 0.49, p < 0.001) to the clinically graded hormonal hirsutism score, while SHBG showed an inverse correlation to it. By contrast, no correlations were found in patients with a serum LH/FSH ratio exceeding 3.0 (n = 15). Various indices for hyperandrogenism were calculated. In patients with lower LH/FSH ratio, T/SHBG, T/SHBG + A/100 and T/SHBG + A/100 +DHEAS/100 showed the best correlation with the clinically scored hair growth. These results show that correlations between hirsutism and hyperandrogenism can be demonstrated.

A method for measurement of free testosterone in premenopausal women involving equilibrium dialysis, chromatography, and radioimmunoassay

A three-step procedure to evaluate the quantity of true free testosterone in female serum was established. Unbound and bound testosterone were first separated by equilibrium dialysis using undiluted serum. Testosterone in dialysate was extracted and separated from cross-reactive steroids by column chromatography and finally quantified with a sensitive radioimmunoassay. The detection limit of the radioimmunoassay was 2.7 pmol/L and the detection limit of the whole method was 6 pmol/L. The intra-assay coefficients of variation of the method were 20.6%, 16.8%, and 9.5% for free testosterone concentrations 12.0 pmol/L, 21.4 pmol/L, and 31.4 pmol/L, respectively. The interassay coefficients of variation were 26.3% and 14.8% for free testosterone concentrations 10.4 pmol/L and 21.3 pmol/L, respectively. Serum free testosterone concentrations were determined in 27 regularly menstruating control women. The mean concentration of actual free testosterone was 10.0 +/- 4.9 pmol/L (mean +/- SD, range from below 6 to 19 pmol/L, n = 27).

Androgen parameters in hirsute women: Correlations with body mass index and age

The recent work was designed for studying the effects of body mass index (BMI) and age on serum hormone values in hirsute women of reproductive years with nontumorous hyperandrogenism. Two hundred patients were studied in the follicular phase of the cycle. The serum levels of testosterone (T), androstenedione (A), or dehydroepiandrosterone sulfate (DHEAS) were not associated with BMI, whereas the T/sex hormone-binding globulin (T/SHBG) ratio was significantly correlated with BMI independently of the effect of age. All serum androgens were significantly negatively correlated with age, and the serum levels were significantly lower in women older than 41 years when compared with women aged 25 years or less. After correcting for the effect of BMI, there was also a negative correlation between the T/SHBG ratio and age. The correlations between serum androgen parameters were calculated. We conclude that already in reproductive years, the age of the patient has modulating effects on serum androgens, and it should be taken into account when studying the hormone profile of hirsute women.

Acute Effect of Exogenous Thyroxine Dose on Serum Thyroxine and Thyrotrophin Levels in Treated Hypothyroid Patients

Acute effects of exogenous thyroxine on the serum thyroxine (T4), free thyroxine (FT4) and thyrotrophin (TSH) levels were studied in 20 hypothyroid women during the long-term maintenance therapy. All patients were euthyroid by clinical and laboratory parameters (T4). In nine patients TSH secretion was totally suppressed (less than 1 mu/l) by the mean dose of 178 micrograms thyroxine. Eleven patients had TSH values greater than 1 mu/l. Their mean thyroxine dose (132 micrograms) was not significantly lower than that which totally suppressed the TSH secretion. Ingestion of thyroxine resulted in maximal T4 and FT4 levels at 2 h after taking the drug. Simultaneously TSH concentration reached its minimum. The results indicate that it is necessary to know when the patient has taken the thyroxine in relation to determination of T4, FT4 and TSH levels in order to make the right decisions about the substitution dose of thyroxine. Thus it is essential to take blood sample for hormone determination before the patient ingests the daily thyroxine dose. However, among hypothyroid patients substituted by thyroxine there are individuals whose TSH secretion is not suppressed by normal T4 and FT4 levels.