Articular cartilage chondrocytes express aromatase and use enzymes involved in estrogen metabolism

Introduction

Sex hormones, especially estrogens, have been implicated in articular cartilage metabolism and the pathogenesis of postmenopausal osteoarthritis. The conversion by aromatase (CYP19A1) of androstenedione into estrone (E1) and of testosterone into 17β-estradiol (E2) plays a key role in the endogenous synthesis of estrogens in tissue.

Methods

We analyzed the expression of aromatase (CYP19A1) in immortalized C-28/I2 and T/C-28a2 chondrocytes, as well as in cultured primary human articular chondrocytes and human articular cartilage tissue, by means of RT-PCR, Western blotting and immunohistochemistry. By means of quantitative RT-PCR and enzyme-linked immunosorbent assay, we also determined whether the aromatase inhibitor letrozole influences estrogen metabolism of cultured chondrocytes in immortalized C-28/I2 chondrocytes.

Results

Aromatase mRNA was detected in both immortalized chondrocyte cell lines, in cultured primary human chondrocytes, and in human articular cartilage tissue. By means of Western blot analysis, aromatase was detected at the protein level in articular cartilage taken from various patients of both sexes and different ages. Cultured primary human articular chondrocytes, C-28/I2 and T/C-28a2, and human articular cartilage tissue reacted with antibodies for aromatase. Incubation of C-28/I2 chondrocytes with 10⁻¹¹ M to 10⁻⁷ M letrozole as an aromatase inhibitor revealed significantly increased amounts of the mRNAs of the enzyme cytochrome P4501A1 (CYP1A1), which is involved in the catagen estrogen metabolism, and of the estrogen receptors ER-α and ER-β. Concomitantly, synthesis of estrone (E1) was significantly downregulated after incubation with letrozole.

Conclusions

We demonstrate that human articular cartilage expresses aromatase at the mRNA and protein levels. Blocking of estrone synthesis by the aromatase inhibitor letrozole is counteracted by an increase in ER-α and ER-β. In addition, CYP1A1, an enzyme involved in catabolic estrogen metabolism, is upregulated. This suggests that articular chondrocytes use ERs functionally. The role of endogenous synthesized estrogens in articular cartilage health remains to be elucidated.     

Identification and Characterization of Cysteinyl-Leukotriene Formation in Tissue Slices from Human Intracranial Tumors: Evidence for Their Biosynthesis Under In Vivo Conditions

In previous studies, it had been shown that human gray and white matter tissue slices have the capacity to synthesize large amounts of cysteinyl-leukotrienes (cysteinyl-LT) in vitro. This study was initiated to investigate cysteinyl-LT formation by tissue slices from intracranial tumors in comparison with cyclooxygenase products such as prostaglandin (PG) F2 alpha and thromboxane (TX) B2. Tissue slices from meningiomas and astrocytomas were found to release large amounts of cysteinyl-LT spontaneously and even higher amounts after ionophore A 23187 stimulation, which could not be accounted for by blood possibly remaining in the tissue slices. Cysteinyl-LT were identified by their immunoreactive characteristics, their biological activity in the guinea pig ileum bioassay, and their retention time on reversed-phase HPLC. With increasing malignancy, astrocytomas were shown to have an increasing biosynthetic capacity for cysteinyl-LT and TXB2 in vitro. In comparison with meningioma patients, malignant astrocytoma patients had an enhanced urinary excretion of LTE4, the major urinary metabolite of cysteinyl-LT in humans, which dropped in level within 7 days after operation by 79%. A correlation exists between the in vivo cysteinyl-LT production in patients with malignant astrocytomas and that observed under basal conditions in the tissue slices in vitro. The results provide evidence that in malignant astrocytoma patients, the tumor tissue produces large amounts of cysteinyl-LT, which may be detected in the patients' urine.     

Does low back pain affect the reliability of postural regulation?

The aim of this study was to examine the intraobserver reliability of a posturographic assessment in patients with low back pain. We investigated 24 symptomatic subjects with defined low back pain (mean: 57.9?years) and a pair-matched control group including 24 asymptomatic persons (mean: 58.1?years). Each participant underwent two measurements on a posturographic device (32?Hz sampling rate) based on the Interactive Balance System (time interval: 7?d). Test procedure consisted of tests on solid ground with eyes open (1) and eyes closed (2). Data analysis included parameters of motor output and a frequency band analysis. Reliability tests were realized using by intraclass correlations (ICC). Coefficients of ICC ranged from 0.36 (95% CI: 0.01–0.73) to 0.94 (95% CI: 0.86–0.97) in both test positions. For 69% (11/16) of the investigated parameters a high level (ICC?>?0.75) of intraobserver reliability was reached. Based on the results, the posturographic measurement system used in this study seems to be appropriate for use in longitudinal study designs in an orthopaedic setting.     

Myositis in Lewis rats induced by the superantigen Staphylococcal enterotoxin A

Molecular Biology Reports - The aetiology of inflammatory myopathies is not clearly known. A predominance of activated Cd8+ T lymphocytes in inflammatory infiltrates has already been detected....     

Olfactory sensitivity of subjects working in odorous environments

The aim of the present study was to investigate whether people with a professional interest in odors also exhibit higher olfactory sensitivity. To this end, we investigated 58 subjects (age 33.6 +/- 11.0 years, mean +/- SD; 55 women) employed in perfume retail outlets and compared their olfactory sensitivity to 58 controls (age 34.6 +/- 9.9 years; 53 women) matched for age, gender and professional activities who did not work in such odorous environments. Olfactory function was assessed using the 'Sniffin' Sticks' test kit which includes tests for n-butanol odor threshold, odor discrimination and odor identification. Subjects working in perfume retail outlets scored higher in odor discrimination tests compared to controls. Working in an odorous environment for a full day had no major effect on general olfactory abilities, as indicated by measures performed at the beginning and end of a working day. Taken together, results from the present study do not support the idea that odorous environments are deleterious to general olfactory function.     

Comparison of intraobserver single-task reliabilities of the Interactive Balance System (IBS) and Vertiguard in asymptomatic subjects

The intraobserver reliabilities of the Interactive Balance System (IBS) and Vertiguard were compared in 30 asymptomatic volunteers. Relative reliability for all IBS single tasks and parameters in the IBS was excellent (intraclass correlation coefficient, ICC ≥0.75). The ICC values ranged from 0.78 to 0.89. The ICC values of the Vertiguard system ranged from 0 to 0.75. The cumulative measure of injury risk of the Vertiguard system was highly reliable.