Effect of steroid hormones on endotoxin-mediated cartilage degradation

Summary Cartilage degradation is a characteristic feature of various types of human arthritis, notably rheumatoid arthritis and osteoarthritis. The influence of glucocorticoid and other steroid hormones on cartilage proteoglycan breakdown was examined in a model system in which breakdown is readily quantified by the release of proteoglycan from cultured bovine nasal cartilage discs. Endotoxin (bacterial lipopolysaccharides) treatment enhanced the depletion of cartilage proteoglycan by 2–3 fold. This was inhibited in a concentration-dependent manner by hydrocortisone (10^–9 to 10^–5M) or other glucocorticoid hormones (dexamethasone, prednisolone, cortisone). Inhibition required the continued presence of the steroid. Removal of hydrocortisone (3 × 10^–7M) after 4 days from endotoxin-treated cultures resulted in the rapid restoration of an endotoxin response, so that proteoglycan release approached maximum levels during a second 4-day culture period. Other C-21 steroid hormones (progesterone, aldosterone) were also inhibitory at 10^–5M, but testosterone and -estradiol showed little influence on endotoxin action. Proteoglycan products of smaller average mol wt (Sepharose CL-2B chromatography), consistent with core protein cleavages, were released from endotoxin-treated cartilage. Cleavage was unaffected by -estradiol, partially blocked by aldosterone and largely prevented by hydrocortisone administration.     

Effect of stress and stress hormones on the hormone (insulin) binding of Tetrahymena

The unicellular Tetrahymena pyriformis GL produce, store and secrete vertebrate‐like hormones. In earlier experiments the effect of different stressors on the hormone levels of Tetrahymena was studied and an elevation of these was found. In the present experiments the hormone binding was investigated, using flow cytometric method. FITC‐insulin binding was elevated after concentrated (5, 10, or 20 mg ml^?1) NaCl or 0.01%, 0.1%, or 0.05% formaldehyde treatment, or after thermal stress (37°C). Serotonin given together with NaCl increased and together with formaldehyde decreased the binding. Histamine always decreased the binding and insulin was indifferent. Four hours after osmotic stress, hormone binding significantly decreased and this was not influenced by hormones. However, 4 h after formaldehyde stress the binding elevated and this was further increased by repeated hormone treatments. The results show that the stress in Tetrahymena provokes an activation of its hormonal system (hormone production and binding), which is differently influenced by exogeneously given hormones. Copyright © 2009 John Wiley & Sons, Ltd.     

Effects of indomethacin on the divisional morphogenesis and cytoskeleton-dependent processes of Tetrahymena

Indomethacin (0.1 mM) causes significantly altered phospholipid synthesis in Tetrahymena and is able to influence the inositol phospholipid signalling system (9). In the present study the effects of indomethacin on the course of cell division, cyclin expression, the cortical microtubular system and on cytoskeleton-dependent processes (motility, phagocytosis) were investigated. As expected from its interference with the synthesis of phospholipids, indomethacin affected Tetrahymena in a number of ways: the structure of the cortical microtubular system became irregular; in many cells the stomatogenesis (development of new oral apparatus) and the development of the fission furrow was not accompanied by elongation of the macronucleus, which is a typical phenomenon of the normal course of mitosis: apparently indomethacin uncouples these phenomena. After indomethacin treatment, the expression of both cyclin A and cyclin B(1) were reduced significantly. The cell growth rate, motility and phagocytotic activity were all considerably reduced. There are probably additional mechanisms responsible for the effect of indomethacin on the systems that control divisional morphogenesis, for microtubule-dependent processes and for the connection between nuclear and cortical alterations during the cell cycle. Effects on protein phosphorylation/dephosphorylation, on cyclin expression and on microtubular functions are probably involved. These possibilities are discussed.     

Direct transcriptional targets of sex steroid hormones in bone

The sex steroid hormones, androgens and estrogens, via their respective nuclear receptors, regulate bone mineral density in humans and mice. Very little is known about the direct targets of the androgen and estrogen receptors in bone cells. First, models of hormone and receptor deficiency in mouse and human bone are discussed. This review then focuses on the direct targets of the receptors in osteoblasts and osteoclasts. A direct target of a NR is defined here as a gene that is regulated by NR binding to the DNA (either through DNA binding or association with a DNA binding protein) at an enhancer or promoter of that gene. The experimental evidence that illustrates androgen and estrogen gene regulation in osteoblasts and osteoclasts will be summarized and compared with the phenotype of the hormones in vivo.     

A preliminary study of steroid reproductive hormones in human hair

Nowadays research and clinical studies of human reproductive endocrinology are generally carried out using human blood reproductive hormone assays. However the acquisition of human blood samples has some shortcomings. In search of new approaches, we paid attention to the fact that progesterone can be detected in cow's hair. Consequently we investigated whether or not steroid hormones are measurable in human hair. The results showed that the levels of steroid hormones in hair are not affected by shampoo and do not significantly vary between different segments of hair (i.e. top, middle and basal segments). The menstrual estradiol and progesterone rhythm of female hair is similar to that of female serum. The ratio of hair estradiol to serum estradiol in the female is 41.2% and that of hair progesterone to serum progesterone is 59.0%; the ratio of hair testosterone to serum testosterone in male is 116%. There are significant correlations between hair and serum steroid hormones of healthy human adult: γ (estradiol)=0.395 (n=20), p<0.05; γ (progesterone)=0.440 (n=22), p<0.025 and γ (testosterone)=0.395 (n=25), p<0.05.     

Hormonal interactions in Tetrahymena: effect of hormones on levels of epidermal growth factor (EGF)

Tetrahymena pyriformiswas treated with insulin, histamine or serotonin for 30 min and epidermal growth factor (EGF) level was studied inside the cells using specific antibodies and flow cytometry as well as confocal microscopy. The EGF concentration was highly significantly elevated after hormone treatment, regardless of the hormone used. EGF was localized mainly in the cortical region (mucocysts) and in vesicles and this localization did not differ in untreated and treated cells. The results call attention to the possibility of interactions between hormones at unicellular level and points to the presence of a hormonal system in Tetrahymena that includes receptors, hormones and signal transduction pathways as well as hormonal interactions. This could be the basis of further evolution to the hormonal system of multicellulars.     

Effects of extremely low concentrations of hormones on the insulin binding of Tetrahymena

FITC-insulin binding to previously hormone-treated Tetrahymena was studied by flow cytometry and confocal microscopy. Hormones produced by Tetrahymena were chosen for study and the hormone concentrations were administered between 10(-6) and 10(-21)M for 30 min. Endorphin, serotonin and insulin significantly reduced the hormone binding however histamine did not influence it at all. Endorphin, serotonin and insulin were significantly effective down to 10(-18)M and the effect of insulin and endorphin suggest a similar mechanism. The results call attention to the efficacy of very low hormone concentrations, which can influence the hormone content (earlier experiments) and receptor binding capacity (present study) of a unicellular organism. This seems to be very important, as in wild (natural) conditions the dilution of signaling materials secreted by a water-living protozoan is very high. In addition, the results point to the selectivity of response, as not all of the hormones that deeply influence other physiological indices (e.g. histamine) have an effect on insulin content or insulin receptors.     

2d:4d, sex steroid hormones and human psychological sex differences

Studies on 2d:4d, the ratio between the second and the fourth digit, as a possible indicator of prenatal androgen exposure, have failed to produce consistent results. This paper analyzes the relation between 2d:4d, sex steroids and well-documented sex differences in characteristics such as depression, dominance, and aggressive (ART) and non-aggressive adolescent risk-taking (NART) in a comparatively large sample of adolescent boys (N = 301, mean age: 14.4 years) and girls (N = 298, mean age: 14.3 years). Boys had on average a lower 2d:4d than girls (F = 42.15; p < 0.001). With respect to boys, controlling for age and pubertal development (PD), a small but marginally significant positive association was found between 2d:4d and total testosterone (TT) (r = 0.11; p < 0.05). In girls a significant association was found between 2d:4d and SHBG (r = 0.18; p < 0.01). However, relationships between 2d:4d and hormones depended on the phase of the menstrual cycle, with 2d:4d being negatively associated with FT (B = − 0.013; p < 0.05) once a positive association between 2d:4d and FT for girls in the mid-cycle group (B = 0.019; p < 0.01) is taken into account. With respect to sex differences in characteristics, we found evidence of a relationship between 2d:4d and depression in boys (r = − 0.14; p < 0.05) but not between 2d:4d and dominance, ART or NART. No relationships were found between 2d:4d and any of these variables in girls.     

Effects of peptides and steroid hormones on cell kinetic parameters of normal human breast tissue in organ culture

Summary Several studies have shown the importance of different hormones in the regulation of mammary tsssue growth. The use of organ culture techniques has shown tremendous value for the knowledge of cell proliferation in human breast tissue. Therefore, the purpose of these studies was to analyze the length of the cell cycle, DNA-labeling index, mitotic index, and growth fraction under the effects of insulin, hydrocortisone, and 17-β estradiol in 5-d organ culture. Normal tissues obtained from patients who underwent breast surgery for benign lesions were individually cultured at 37°C (95% air:5% CO₂ in Medium 199). Autoradiographic studies indicated that the hormones shortened the length of cell cycle of normal breast tissue in 5-d organ cultures. From the growth fraction studies we concluded that the hormones may have stimulated the cells to reenter the cell cycle from G₀ because these values were increased by the hormones used. Estrogen can alter the S phase duration with a consequent increase in the rate of DNA synthesis which may explain the high DNA-labeling index observed in the present studies. Supported by Public Health Service grant CA38921 from the National Cancer Institute, Bethesda, MD, and by an Institutional grant from the United Foundation of Greater Detroit.     

Cytochrome P450 3A9 catalyzes the metabolism of progesterone and other steroid hormones

The catalytic requirements and the role of P450 3A9, a female-specific isoform of CYP3A from rat brain, in the metabolism of several steroid hormones were studied using recombinant P450 3A9 protein. The optimal steroid hormone hydroxylase activities of P450 3A9 required cholate but not cytochrome b₅. P450 3A9 was active in the hydroxylation reactions of testosterone, androstenedione, progesterone and dehydroepiandrosterone (DHEA). No activity of P450 3A9 toward cortisol was detectable under our reconstitution conditions. Among all the steroid hormones examined, female-specific P450 3A9 seemed to catalyze most efficiently the metabolism of progesterone, one of the major female hormones, to form three mono-hydroxylated products, 6-, 16-, and 21-hydroxyprogesterone. Our data also showed that P450 3A9 can catalyze the formation of a dihydroxy product, 4-pregnen-6, 21-diol-3, 20-dione, from progesterone with a turnover number, 1.3 nmol/min/nmol P450. Based on the V_max/K_m values for P450 3A9 using either 21-hydroxprogesterone or 6-hydroxyprogesterone as a substrate, 4-pregnen-6, 21-diol-3, 20-dione may be formed either by 6-hydroxylation of 21-hydroxprogesterone or 21-hydroxylation of 6-hydroxyprogesterone. As a major isoform of CYP3A expressed in rat brain, the activities of P450 3A9 toward two major neurosteroids, progesterone and DHEA suggested a possible role for P450 3A9 in the metabolism of neurosteroids.     

Increase of Growth Rate and Phagocytic Activity of Tetrahymena Induced By Pseudomonas Lectins

Galactosephilic and mannosephilic lectins from Pseudomonas aeruginosa interact with Tetrahymena pyriformis GL. Specific adsorption of these lectins onto the Tetrahymena can be shown by inhibition of hemagglutination and by peroxidase binding to the cells mediated by the mannosephilic lectins. Interaction with the lectins does not agglutinate the protozoa even after immobilization by Na fluoride, formaldehyde, and glutaraldehyde or after papain treatment. However, inclusion of the lectins in the growth medium increases the growth rate of Tetrahymena and their presence in the medium supplied to starved ciliates increases phagocytosis of Chinese ink and vacuolization.     

Steroid hormones and plant growth and development

The occurrence of steroid hormones in plants is briefly reviewed. Their effects on plant growth, development and flowering are also considered.     

Pubertal timing, hormones, and body composition among adolescent Turkana males

The Turkana, like other East African pastoral groups, are known for their tall adult stature, achieved despite a blunted growth spurt during adolescence and continued growth into the early 20s. To investigate the hormonal mechanisms associated with the pattern of slow and continued adolescent growth, we collected data on hormonal status, height, weight, and trunk skinfolds and ethnographic self-reports of testicular maturation in a cross-sectional sample of 35 nomadic and 37 settled Turkana males aged 14-24. Hormonal determinations included testosterone (T), sex hormone-binding globulin (SHBG), and dehydroepiandrosterone (DHEA) in blood, in addition to urinary DHEA. Self-reports of testicular maturation showed no difference between settled and nomadic subpopulations. However, nomadic boys exhibited significantly higher levels of T, DHEA, and SHBG. Of all the hormones, only SHBG showed a significant relationship with age. Multiple regression models show blood T and SHBG to be significant independent predictors of achieved height as well as weight, controlling for age. Our results suggest that onset of puberty is substantially delayed among Turkana males, and that bioavailable T is related to growth in stature during adolescence. We suggest that SHBG acts to mediate the effects of energy availability on adolescent growth in this energetically limited population. Our findings may also have implications for understanding adolescent growth among Homo erectus.     

Chemotactic activity of oligopeptides containing an EWS motif on Tetrahymena pyriformis: the effect of amidation of the C-terminal residue

Chemotactic properties of 3-7-mer peptides containing an EWS motive and their peptide amides synthesized and characterized by us were investigated in Tetrahymena pyriformis GL model. Analysis of the peptide acids shows that SEWS possesses exceptionally strong (660%+/-21; 430%+/-18) chemoattractant ability at 10(-12) and 10(-11) m respectively. The shorter peptide (EWS) possesses chemorepellent activity, while longer peptides display neutral (WSEWS) or moderate chemoattractant (EWSEWS and GEWSEWS) chemotactic ability. Amidation of the C-terminus can significantly modify the character of peptides: it points to the conclusion that a free alpha-COOH group at this position is required for the high efficiency of SEWS, while in the shorter (EWS) and longer peptides (WSEWS and EWSEWS) amidation can result in chemoattractant ligands. Evaluation of the structure-function relationship of these compounds establishes the significance of Glu (E) with its high surface-exposed area and negatively-charged side chain. The high discriminative ability and good chemotactic responsiveness of Tetrahymena support the theory that a chemotactic signalling mechanism working in higher levels of phylogeny is a well conserved and inducible one even in protozoa.     

Insulin antagonizes the phagocytosis stimulating action of histamine in Tetrahymena

Histamine increased specifically the phagocytic activity of the unicellular Tetrahymena, whereas insulin had no influence on it. Insulin antagonized the phagocytosis stimulating action of histamine after simultaneous exposure and after preexposure two days earlier as well, although in the latter case to a lesser degree. Double exposure to a combination of histamine + insulin didn't influence the phagocytic activity at all, demonstrating the histamine antagonizing effect of insulin in this model.     

A general response to stressors by the unicellular Tetrahymena: effect of stress on the hormone levels

Hormone (ACTH, endorphin, serotonin and T(3)) content of the unicellular Tetrahymena was studied by using immunocytochemical and flow cytometry methods. The cells were previously treated with different concentrations of salts (NaCl or KCl for 1 h), were kept in low (+4 degrees C) or high (+37 degrees C) temperature for 1 h and were treated with formaldehyde or ethanol. High concentrations of salts (20 mg ml(-1) medium), all levels (0.1, 0.05, 0.01%) of formaldehyde and high temperature almost doubled the hormone contents and 0.1% alcohol also significantly elevated them. The experiments call attention to the presence of a general adaptation syndrome (GAS)-like phenomenon at a unicellular level and points to the possibility of deduction of GAS to a low level of phylogeny.     

The detection and monitoring of early pregnancy in the vervet monkey (Cercopithecus aethiops) with the use of ultrasound and correlation with reproductive steroid hormones

Twenty early pregnancies were diagnosed and monitored in vervet monkeys by ultrasonography. Non-gravid uteri became increasingly echogenic from cycle days 7 to 26. The first definite sign of pregnancy was a gestational cavity of 2 mm (+/- 0.80) at 33.0 (+/- 1.48) days menstrual age, which was also used to date all subsequent features. Earlier signs, such as an endometrial line swelling or endometrial 'pregnancy' ring, as reported for other non-human primate species, could not be reliably and consistently used to diagnose pregnancy in vervet monkeys. A rapid increase of the gestational cavity size from days 37 to 49 corresponded closely to a rapid increase in plasma progesterone concentration from day 39 to 49. The first yolk sac was recognizable at 38.0 days (+/- 3.10) and measured 3.3 mm (+/- 0.40) in diameter. A heart beat could be detected at 45.5 (+/- 1.73) days and the size of the first measurable embryo at 35 days was 2 mm. The dating of most features was within the range reported for other non-human primate species.