Th1, Th2 and Th17 lymphocytes in autoimmune thyreopathies.

Among the most common organ-specific autoimmune diseases are autoimmune thyreopathies. We have focused on the importance of Th1, Th2 and Th17 lymphocytes in autoimmune thyroiditis (AT). The cohort consisted of 136 treated patients in the full clinical course of AT (24 men, mean age 41.0±16.8 years and 112 women, mean age 44.6±17.6 years). The control group consisted of 17 healthy men (mean age 44.0±5.0 years). Box-Cox transformation of the data, t-tests and Pearson correlation analysis were used for statistical calculations. Lymphocyte subpopulations were determined by flow cytometry. We found statistically significant correlations between Th and Tc lymphocytes (r = -0.5605, p = 0.0000), total T and B lymphocytes (r = -0.4877, p = 0.0000), Th1 and Th17 lymphocytes (r = 0.4346, p = 0.0000), Tc and Th1 lymphocytes (r = 0.4124, p = 0.0000), IRI and Th1 lymphocytes (r = -0.4076, p = 0.0000), total T lymphocytes and NK cells (r = -0.8175, p = 0.0000), memory Th and Th1 lymphocytes (r = 0.7982, p = 0.0000), naive Th and Th1 lymphocytes (r = -0.7995, p = 0.0000), Tc lymphocytes and NK cells (r = -0.4014, p = 0.0000), Tc and total T lymphocytes (r = 0.4551, p = 0.0000), Th and total T lymphocytes (r = 0.4135, p = 0.0000). The determination of lymphocyte subpopulations is an aid in the diagnosis and treatment of autoimmune diseases, helps to clarify the clinical manifestations of the disease and can complement the interpretation of commonly determined autoantibodies. It can help determine whether the phase is destructive (Th1, Th17, Tc lymphocytes) or protective (Th2 lymphocytes, antibodies).

[Psycho-immuno-endocrinology of the thyroid gland]. / Psycho-imuno-endokrinologie stítné zlázy.

Historically endocrinologists and psychiatrists are aware that disturbances in thyroid disease in beginning or even in clinically intensified states of thyrotoxicosis or hypothyroidism exhibit pathological mental manifestations, masking or potentiating the underlying disease. Immune system disorders cause thyroid organ-specific autoimmune process. This autoimmune thyroid disease binds with a number of disorders in both endocrine or non-endocrine organs. This appears in vascular, neurological, skin, connective tissue, gastrointestinal tract and mental pathology. These disorders are part of autoimmune polyglandular syndromes (APS) type I -III, especially the APS type III. Originally it was assumed that these mental disorders are caused by direct exposure to excess or deficiency of thyroid hormones. Recently, however, it appears that these psycho-immune-endocrine disorders have common etiologic mechanisms of formation and on cellular and molecular level they involve similar, if not in some cases, common mechanisms.Key words: antithyroid peroxidase antibody - autoimmune polyglandular syndrome type I., II., III. - autoimmune thyroid disease - bipolar disorder - depression - Hashimotos encephalopathy - postpartum psychosis - psycho-immuno-endocrinology - schizophrenia.

Trace elements and bone health.

The importance of nutrition factors such as calcium, vitamin D and vitamin K for the integrity of the skeleton is well known. Moreover, bone health is positively influenced by certain elements (e.g., zinc, copper, fluorine, manganese, magnesium, iron and boron). Deficiency of these elements slows down the increase of bone mass in childhood and/or in adolescence and accelerates bone loss after menopause or in old age. Deterioration of bone quality increases the risk of fractures. Monitoring of homeostasis of the trace elements together with the measurement of bone density and biochemical markers of bone metabolism should be used to identify and treat patients at risk of non-traumatic fractures. Factors determining the effectivity of supplementation include dose, duration of treatment, serum concentrations, as well as interactions among individual elements. Here, we review the effect of the most important trace elements on the skeleton and evaluate their clinical importance.

Influence of metals on cytokines production in connection with successful implantation therapy in dentistry.

OBJECTIVES: In most of patients in need of implantation treatment in the oral cavity, implants heal well, nevertheless, there are some individuals, in whom titanium implants fail for reasons, which remain unclear. DESIGN: The aim of our study was to determine if there is a difference between metal influenced IL-1ß, IL-4, IL-6, TNF-α and IFN-γ cytokines production in patients with successfully healed implants compared to those, whose implant therapy was unsuccessful. SETTING: The two study groups included 12 patients with failed dental titanium implants and 9 patients with successfully healed implants. In the subjects, cytokine production was established after lymphocyte cultivation with mercury, nickel and titanium antigens. RESULTS: IL-1ß levels were significantly increased in all patients after stimulation with titanium and in patients with accepted implants compared to patients with failed implants after the stimulation with mercury and titanium. Titanium caused significantly increased IL-6 production in all patients. TNF-α and IFN-γ levels were also significantly increased after the stimulation with titanium. Significantly increased TNF-α levels were found in patients with accepted implants as compared to patients with failed implants. CONCLUSIONS: Increased production of IL-1ß a IL-6 cytokines in reaction to titanium and increased production of TNF-α and IFN-γ cytokines in reaction to mercury, which is very often present in the form of amalgam in the oral cavity of persons in need of implant therapy, can play an important role in immune reactions during implant healing process. In patients with failed titanium implants, decreased production of these cytokines may participate in implant failure.

Relations of prediabetes and type 2 diabetes to the thyroid cancer.

OBJECTIVE: Patients with type 2 diabetes (T2DM) generally experience a higher incidence of cancer. However, the association between T2DM and thyroid cancer is inconclusive. METHODS: Case-control prospective study, 722 patients were screened for T2DM and prediabetes (PDM) and underwent thyroid ultrasound and biochemical tests. The patients were assigned to groups of PDM (n=55), T2DM (n=79) or a non-diabetes group (NDM) (n=588). Fine needle aspiration biopsy was carried out in 263 patients. Histological examinations were done for 109 patients after surgery, with findings of 52 benign (BS) and 57 malignant tumors (MS). RESULTS: 33 % of patients with T2DM and especially PDM were newly diagnosed by our screening: 6.5 % with T2DM and 72 % with PDM, respectively. The percentage of thyroid cancers did not significantly differ between the groups (χ2 test=0.461; p=0.794). Relevant positive thyroid predictors for T2DM (t-statistic=25.87; p<0.01) and PDM (21.69; p<0.01) contrary to NDM (-26.9; p<0.01) were thyroid volume (4.79; p<0.01), thyroid nodule volume (3.25; p<0.01) and multinodular thyroid gland (4.83; p<0.01), while negative relevant predictors included the occurrence of autoimmune thyroid disease (AITD) (-2.01; p<0.05). CONCLUSION: In general, we did not observe an increased risk for thyroid cancer in the diabetic and prediabetic groups in comparison to controls, in spite of well-established increased risk for other malignancies. Structural and benign changes such as larger and multinodular thyroid glands, in comparison to autoimmune thyroid disease, are present more often in diabetics.

Removal of dental amalgam decreases anti-TPO and anti-Tg autoantibodies in patients with autoimmune thyroiditis.

OBJECTIVES: The impact of dental amalgam removal on the levels of anti-thyroid peroxidase (anti-TPO) and anti-thyroglobulin (anti-Tg) antibodies was studied in patients with autoimmune thyroiditis (AT) with and without mercury allergy. METHODS: Thirty-nine patients with AT were tested by an optimized lymphocyte proliferation test MELISA for allergy (hypersensitivity) to inorganic mercury. Patients were divided into two groups: Group I (n = 12) with no hypersensitivity to mercury and Group II (n = 27) with hypersensitivity to mercury. Amalgam fillings were removed from the oral cavities of 15 patients with hypersensitivity to mercury (Group IIA) and left in place in the remaining 12 patients (Group IIB). The laboratory markers of AT, anti-TPO and anti-Tg autoantibodies, were determined in all groups at the beginning of the study and six months later. RESULTS: Compared to levels at the beginning of the study, only patients with mercury hypersensitivity who underwent amalgam replacement (Group IIA) showed a significant decrease in the levels of both anti-Tg (p=0.001) and anti-TPO (p=0.0007) autoantibodies. The levels of autoantibodies in patients with or without mercury hypersensitivity (Group I and Group IIB) who did not replace amalgam did not change. CONCLUSION: Removal of mercury-containing dental amalgam in patients with mercury hypersensitivity may contribute to successful treatment of autoimmune thyroiditis.

Autoimmune thyroiditis and Helicobacter pylori--is there a connection?

OBJECTIVES: In this study we examined the anti-Helicobacter pylori (anti-H. pylori) antibodies in patients with autoimmune thyroiditis, with and without different polyglandular involvement, and in healthy controls. MATERIAL & METHODS: Patients with autoimmune thyroiditis (AT) were divided into three groups: Group A: 23 patients with isolated AT, Group B: 30 patients with AT as a part of polyglandular activation of autoimmunity, and Group C: 7 patients with AT as a part of autoimmune polyglandular syndrome type II. Thirty healthy individuals served as controls (Group D). Anti-H. pylori antibodies were determined first by ELISA for classes IgG, IgA, and IgM, and subsequently by immunoblot for classes IgG and IgA. RESULTS: ELISA: The number of patients with IgA antibodies in Group A (39%) and Group B (30%) differed significantly from controls (7%, p<0.05). Immunoblot: Anti-CagA antibodies were found in 13% of patients in Group A, 7% of Group B, 0% of Group C, and 20% of Group D. A higher seroprevalence, as compared to controls, was found for IgG to the VacA (p=0.01), 30 kDa (p=0.001), and 17 kDa (p=0.008) antigens in Group A and for IgG to the 30 kDa antigen in Group C (p=0.037). A significantly higher seroprevalence, as compared to controls, was likewise found for IgA to the 17 kDa antigen in Group A (p=0.015). CONCLUSIONS: A different distribution of antibodies to H. pylori antigens was found in patients with isolated AT compared to patients with AT coupled with a polyglandular syndrome.

Immunomodulatory cytokines in human seminal plasma correlate with immunomodulatory steroids.

7alpha-Hydroxy-dehydroepiandrosterone and its 7beta-hydroxyepimer, which act as local immunomodulatory agents, dehydroepiandrosterone, cortisol, and major androgens, together with four cytokines-interleukins 2, 4, 10, and IFN-gamma, reflecting the activity of TH1 or TH2 cells present in semen, were measured in seminal plasma from 35 male donors. Cortisol, dehydroepiandrosterone, its sulfate, 7-hydroxy-dehydroepiandrosterone epimers, testosterone, and estradiol were also measured in their blood serum. Steroids and interleukins in semen as well as serum steroids and seminal interleukins were mutually correlated to find out whether a relationship between immunomodulatory steroids and cytokines influencing the immune environment does exist. A highly significant (P<0.001) positive correlation was found between seminal 7beta-hydroxy-dehydroepiandrosterone and IFN-gamma, while a negative correlation was found between cortisol and IL-10. Highly significant positive correlations were also found between serum 7alpha-hydroxy-dehydroepiandrosterone and seminal IFN-gamma and between serum 7beta-hydroxy-dehydroepiandrosterone and seminal IL-2, while a negative correlation was found between serum dehydroepiandrosterone and seminal IL-10. Different and in some instances, even contradictory findings concerning the influence of dehydroepiandrosterone and cortisol on TH1 and TH2 cytokines were observed in seminal plasma as compared to those found by others in serum. The differences can be ascribed to the different environments of mucosal and systemic immunity. Correlations between the levels of steroids and cytokines in seminal plasma did not always correspond to the correlations between given cytokines and hormones in sera. The results, however, are in agreement with our recent finding of an autonomous production of these steroids in the male reproductive tract.

Simple plasma work-up for a fast chromatographic analysis of homocysteine, cysteine, methionine and aromatic amino acids.

Simplified sample workup obviating protein precipitation and eluent evaporation commonly employed in earlier reports using chloroformate-mediated derivatization of aminothiols prior to mass spectrometric (MS) detection is presented. The reduction of disulfides in plasma is accomplished with dithiothreitol within minutes. A simultaneous derivatization with ethyl chloroformate (ECF) and extraction of derivatives into organic phase takes place within seconds. Along with S-amino acids, also aromatic amino acids can be determined during a 5-min run. Gas chromatography with flame ionization detection (GC-FID) proved to be sensitive enough to reach plasma homocysteine levels. A prerequisite for a reliable quantitation was fulfilled under the given conditions. Intra-assay precision was <5%, recoveries from spiked plasma complete (101.2%), detection and quantitation limits for homocysteine came to <1 and 3 micro mol/l. Our results were in full agreement with those obtained by liquid chromatography (r=0.999 for homocysteine and 0.987 for cysteine), and were close to two homocysteine immunoassays (r=0.991 and 0.939, respectively).

Serum levels of angiogenic growth factors in patients with thyroid gland tumors and parathyroid adenoma.

OBJECTIVE: Angiogenic growth factors bFGF and VEGF ensure vascularisation of the growing tumor tissue. We decided to investigate their peripheral serum concentrations in patients with thyroid gland adenoma and papillary carcinoma and with parathyroid adenoma. We wanted to find the possible serum marker of these tumor diseases. METHODS: 28 patients with thyroid gland tumor (14x adenoma, 14x papillary carcinoma) and 12 patients with parathyroid gland adenoma. Growth factors serum levels were measured by ELISA method. RESULTS: We found significantly higher serum levels of bFGF in both groups of patients with thyroid adenoma (4.93 +/- 3.42 ng/ml) and papillary carcinoma (5.69 +/- 5.58 ng/ml) compared to the healthy population (1.47 +/- 1.77 ng/ml). There were no significant differences of VEGF serum levels between all examined groups of patients (adenoma 213 +/- 197, papillary carcinoma 210 +/- 179, healthy 227 +/- 231 pg/ml). We found significantly higher serum levels of bFGF in patients with parathyroid gland adenoma (7.59 +/- 9.12 ng/ml) compared to those in healthy people (1.47 +/- 1.77 ng/ml). CONCLUSIONS: Higher bFGF serum concentrations in patients with thyroid and parathyroid tumors are in accordance with their immunohistochemical tissue levels described in the literature. Not so in VEGF. bFGF may be a serum marker of thyroid and parathyroid neoplasms.

Thyroid surgery in children and adolescents.

Authors present 114 children and adolescents of the total 3865 patients that underwent surgery on the Department of Otorhinolaryngology and Head and Neck Surgery, 1st Faculty of Medicine, Charles University, Prague, Department of Otorhinolaryngology of IPVZ, Prague because of a thyroid disease in years 1991-2000. The male/female ratio in the group was 1:6.5, 16 (13.9%) males and 99 (86.1%) females. For non-malignant disease, the male/female ratio was 1:6.5 and for malignant disease, it was 1:5.4. Thyroid surgery was indicated for non-malignant disease in 82 (71.9%) cases--11 males and 71 females. Most frequently it was indicated because of Graves-Basedow and Hashimoto toxicosis--in 39 children (34.2%), in 5 children (4.4%) for toxic adenoma. Because of the polynodular goiter surgery was indicated in 13 cases (11.4%), in 22 cases (19.3%) for mononodular goiter. Three girls (2.6%) were recommended for surgery because of Hashimoto thyreoiditis and for mechanic syndrome in diffuse goiter one girl (0.9%). Malignant tumor was found in 32 children (28.1%)--5 males and 27 females. Most frequent histological type was papillary cancer--in 25 cases (21.9%). Follicular cancer was diagnosed in four cases (3.5%) and medullar cancer in three cases (2.6%). TNM classification in children and adults is summarized in Table 1. Surgery was carried out in two children with distant metastases (lung localization). In both children the pre-disease stage lasted more than 12 months. The first manifestation of the disease was a finding of enlarged and palpable nodes on the neck. The authors summarize indications for surgery of thyroid diseases in children and adolescents. A treatment of the thyroid gland is a teamwork. Indication for surgery is carried out by a pediatric endocrinologist in cooperation with a surgeon specialized in thyroid surgery. The possible identification of the recurrent laryngeal nerve by a surgery microscope is an unquestionable advance. Children and adults should be centralized into health-care centers capable not only of surgery, but also of endocrinology care. In children with a malignant disease there is a need for cooperation with a pediatric oncologist and a following oncological treatment on a nuclear medicine department. The authors summarize the information about today's thyroid surgery care possibilities for children and adolescents. They point out the differences in indications and the decision about extent of surgery in children and adolescents.

Serum levels of growth factors HGF (Hepatocyte Growth Factor), TGFbeta1 (transforming growth factor beta1) and IGF-I (Insulin Like Growth Factor I) in parathyroid tumors.

INTRODUCTION: HGF (Hepatocyte Growth Factor), TGFbeta1 (Transforming Growth Factor beta1) and IGF-I (Insulin Like Growth Factor I) are cytokines that are involved in the parathyroid tumors formation and growth. We tried to determine, if there are changes and relationships in the production of these cytokines by tumor cells of parathyroid tumors. MATERIAL AND METHODS: We determined concentrations of HGF, TGFbeta1 and IGF-I in serum from peripheral blood of 16 patients with parathyroid adenoma and of 8 patients with parathyroid secondary hyperplasia before and after parathyroidectomy. Results were compared with serum levels in healthy people. RESULTS: Both preoperative and postoperative HGF serum levels in patients with parathyroid adenoma and secondary hyperplasia are significantly higher than in healthy people. Preoperative and postoperative serum levels of TGFbeta1 in parathyroid adenoma and postoperative TGFb1 serum levels in parathyroid secondary hyperplasia are higher, compared with those in the healthy population and in parathyroid secondary hyperplasia preoperatively. There are no significant differences of IGF-I serum levels among the all investigated groups of patients. CONCLUSIONS: Changes in the growth factors production by parathyroid tumor cells are reflected by their concentrations in peripheral blood. The elevation of HGF serum levels in patients with parathyroid adenoma and hyperplasia can be explained by very high HGF production by tumor cells. Nevertheless, there is no decrease of HGF serum levels after the parathyroidectomy. That may be the result of the extratumoral production of this cytokine. Also TGFbeta1 and IGF-I serum levels indicate high possibility of the extratumoral production of these cytokines. Higher postoperative IGF-I serum levels (but not significantly) in parathyroid secondary hyperplasia are in accordance with its bone production.

Mercury and nickel allergy: risk factors in fatigue and autoimmunity.

This study examined the presence of hypersensitivity to dental and environmental metals in patients with clinical disorders complicated with chronic fatigue syndrome. Three groups of patients were examined through medical history, dental examination, and by using a modified test of blast transformation for metals-MELISA(R). The three groups consisted of the following: 22 patients with autoimmune thyroiditis with or without polyglandular autoimmune activation; 28 fatigued patients free from endocrinopathy; and 22 fatigued professionals without evidence of autoimmunity. As controls, a population sample or 13 healthy subjects without any evidence of metal sensitivity was included. Healthy controls did not complain of marked fatigue and their laboratory tests did not show signs of autoimmunity and endocrinopathy. We have found that fatigue, regardless of the underlying disease, is primarily associated with hypersensitivity to inorganic mercury and nickel. The lymphocyte stimulation by other metals was similar in fatigued and control groups. To evaluate clinical relevance of positive in vitro findings, the replacement of amalgam with metal-free restorations was performed in some of the patients. At a six-month follow-up, patients reported considerably alleviated fatigue and disappearance of many symptoms previously encountered; in parallel, lymphocyte responses to metals decreased as well. We suggest that metal-driven inflammation may affect the hypothalamic-pituitary-adrenal axis (HPA axis) and indirectly trigger psychosomatic multisymptoms characterizing chronic fatigue syndrome, fibromyalgia, and other diseases of unknown etiology.

Steroid profiling in pregnancy: a focus on the human fetus.

In this review we focused on steroid metabolomics in human fetuses and newborns and its role in the physiology and pathophysiology of human pregnancy and subsequent stages of human life, and on the physiological relevance of steroids influencing the nervous systems with regards to their concentrations in the fetus. Steroid profiling provides valuable data for the diagnostics of diseases related to altered steroidogenesis in the fetal and maternal compartments and placenta. We outlined a potential use of steroid metabolomics for the prediction of reproductive disorders, misbalance of hypothalamic-pituitary-adrenal axis, and impaired insulin sensitivity in subsequent stages of human life. A possible role of steroids exhibiting a non-genomic effect in the development of gestational diabetes and in the neuroprotection via negative modulation of AMPA/kainate receptors was also indicated. Increasing progesterone synthesis and catabolism, declining production of tocolytic 5ß-pregnane steroids, and rising activities of steroid sulfotransferases with the approaching term may be of importance in sustaining pregnancy. An increasing trend was demonstrated with advancing gestation toward the production of ketones (and 3ß-hydroxyl groups in the case of 3α-hydroxy-steroids) was demonstrated in the fetus on the expense of 3α-hydroxy-, 17ß-hydroxy-, and 20α-hydroxy-groups weakening in the sequence C17, C3, and C20. There was higher production of active progestogen but lower production of active estrogen and GABAergic steroids with the approaching term. Rising activities of placental CYP19A1 and oxidative isoforms of HSD17B, and of fetal CYP3A7 with advancing gestation may protect the fetus from hyperestrogenization. This article is part of a Special Issue entitled 'Pregnancy and Steroids'.

Steroid metabolome in plasma from the umbilical artery, umbilical vein, maternal cubital vein and in amniotic fluid in normal and preterm labor.

The boost in placental production of CRH in late pregnancy is specific for human. CRH receptors are expressed in the fetal zone of the fetal adrenal (FZFA). Hence, we evaluated the associations between the steroid metabolome and gestational age (GA). The levels of 69 steroids and steroid polar conjugates such as 3beta-hydroxy-5-ene steroids (3betaOH5S), 3-oxo-4-ene steroids (3O4S), progesterone 5alpha/beta-reduced metabolites, 20alpha-hydroxy-metabolites of C21 steroids, C19 5alpha/beta-reduced metabolites, 7alpha/beta-hydroxy-metabolites of 3betaOH5S, estrogens and 16alpha-hydroxy-metabolites of 3betaOH5S and 3O4S, were measured by GC-MS in plasma from the umbilical artery (UA), umbilical vein (UV), and maternal cubital vein (MV) and in amniotic fluid (AF) in 12 women at normal labor and 38 women at preterm labor due to pathologies unrelated to steroid status. Using multivariate regression, prediction models for GA were completed for the individual body fluids. The conjugated 3betaOH5S (the key products of the FZFA), estrogens, some polar conjugates of progesterone 5alpha/beta-reduced metabolites and some steroid 7alpha/beta- and 16alpha-hydroxy-metabolites showed strong positive correlations with the GA. The predictivity decreased in the following sequence UV (R=0.950), UA (R=0.945), MV (R=0.895), and AF (R=0.891). Although the predictivity of steroids in maternal blood was slightly less effective when compared with the UV and UA, it was the best solution for further practice.

Short-term effect of soy consumption on thyroid hormone levels and correlation with phytoestrogen level in healthy subjects.

OBJECTIVE: Since soy isoflavones may influence the thyroid hormone feedback system by interference with their biosynthesis, secretion and metabolism, we tested whether their controlled shortterm consumption affects thyroid function. METHODS: Eighty six volunteers--university students (32 males and 54 females) were eating unprocessed boiled natural soybeans (2 g/kg body weight/day) for 7 consecutive days. Thyrotropin, free thyroid hormones, antibodies to thyroid peroxidase and to thyroglobulin, and actual levels of unconjugated major soy phytoestrogens, daidzein and genistein, were measured in sera collected before, at the end and one week after finishing soy meal consumption. RESULTS: Both phytoestrogens increased significantly (p<0.0001) at the end of soy-diet and fell down after its termination nearly back to the initial values. No significant changes were found in female group, while in males a significant transitory increase of thyrotropin (p<0.0001) was recorded. When actual levels of phytoestrogens were related to thyroid parameters, the only significant correlations were found between basal levels of daidzein and thyrotropin, daidzein and antithyroglobulin at the end of soy consumption in males, and between daidzein and free thyroxine at the end of the soy ingestion in females. CONCLUSION: Though only modest and transitory effects on thyroid parameters occurred after controlled short-term soy consumption, some actual thyroid hormone parameters do correlate with actual isoflavone levels.

Novel and traditional biomarkers of bone turnover in postmenopausal women.

Urinary markers of bone collagen degradation such as N-terminal telopeptide, deoxypyridinoline, hydroxyproline and proline-hydroxyproline dipeptide as a novel marker were measured in both morning and daily urine samples collected from 40 postmenopausal women on 10 consecutive days. In addition, osteocalcin, bone alkaline phosphatase and procollagen C-terminal propeptide were analyzed in one serum sample from each of the women. DXA densitometry was employed to differentiate between osteopenic and osteoporotic women. Spearman rank correlation, factor analysis and receiver-operating characteristics were computed to evaluate the results. The highest correlation with a single common factor was found between telopeptide assayed in daily urine, followed by the dipeptide and hydroxyproline assayed in morning urine. These markers also showed high sensitivity for the diagnosis of osteoporosis, while osteocalcin had a high sensitivity for the diagnosis of bone disease. The concentrations of telopeptide, dipeptide and osteocalcin differed significantly between the osteopenic and osteoporotic subjects. Because of the ease of non-hydrolyzed urine processing, the speed of chromatographic analysis and the negligible costs, the dipeptide assay opens new possibilities in the field.