Mitogenic effects of growth hormone in cultured human fibroblasts. Evidence for action via local insulin-like growth factor I production.

We examined human growth hormone's (hGH) effect on mitogenesis in cultured human fibroblasts, and the role of local insulin-like growth factor I (IGF-I). With 0.5% human hypopituitary serum (HPS), hGH increased thymidine incorporation (TI) over serum-free medium dose responsively, with half-maximal effect at 10 ng/ml (0.5 nM) (hGH 127 +/- 8.8%; IGF-I 107 +/- 1.7% [SEM]) (n = 10). Similarly, with 0.5% HPS, hGH and IGF-I increased cell replication by 172 +/- 8.2% and 169 +/- 25%, respectively (n = 4). Specific IGF-I monoclonal antibody (Sm1.2) dose dependently blunted TI stimulated by 10 ng/ml hGH or IGF-I (at 1:1000, 38 +/- 6.5% and 30 +/- 14% reduction, respectively). Sm1.2 also reduced cell replication by both 10 ng/ml hGH and IGF-I, respectively, to 32% and 42% of stimulated values. Dexamethasone (0.1 microM) synergistically enhanced TI by both IGF-I and hGH. A 28-h time course for TI showed that hGH stimulated a similar peak to IGF-I, lagging in its effect by 4-10 h. We have provided further evidence that hGH stimulates growth of cultured human fibroblasts via local IGF-I production, consistent with IGF-I's paracrine-autocrine role.

The ontogeny of growth hormone receptors in the rabbit tibia.

To address the question of the mode of action of GH in stimulating longitudinal bone growth, we have used a panel of anti-GH receptor monoclonal antibodies to demonstrate GH receptors in the rabbit tibia and have studied the ontogeny of these receptors. In the neonate, receptors were localized in the hypertrophic zone between the cartilage canals, a region that develops into a secondary ossification center. In support of this finding, receptors were also localized on monolayer cultures of human infant costal chondrocytes. In 20- and 50-day-old rabbits, receptors were localized on reserve and proliferative chondrocytes in the growth plate. In 50- and 130-day-old rabbits receptors were localized on proliferative chondrocytes in the condylar cartilage. In older (180-day-old) rabbits with closed growth plates, GH receptors could not be detected, even in condylar cartilage. These results support the case for revision of the somatomedin hypothesis to accommodate a direct interaction between GH and receptors on epiphyseal chondrocytes.

Visual demonstration of growth hormone receptors on human growth plate chondrocytes.

The sites of action of GH in the human infant remain unclear; recent evidence in animals suggests direct actions on growth plate and other tissues. We have used a monoclonal antibody recognizing the human GH receptor to visually identify and localize GH receptors in the human infant growth plate. Sternochondral cartilage was obtained at postmortem from infants dying of sudden infant death (n = 20), and either decalcified, fixed, and cut into longitudinal sections or digested with collagenase for monolayer culture of chondrocytes. Sections of cultured chondrocytes were stained immunocytochemically with a monoclonal antibody recognizing human GH receptor (MAb 263), using an avidin-biotin system. Sternochondral cartilage was also obtained at operation from adolescents undergoing sternochondroplasty. In infant tissue, GH receptor was identified in sections in chondrocytes of the proliferative and hypertrophic layers, in perichondrium, in osteocytes in new bone, and in hemopoietic precursor cells in marrow. Cultured chondrocytes showed heterogeneous staining for GH receptor. With prolonged culture from 5-8 days, the pattern of staining changed from individual cells to groups of cells. [125I]Human (h)GH showed specific binding to chondrocyte monolayer (0.6 +/- 0.3%), confirmed visually on emulsion autoradiography. In support of specificity of MAb263, it was able to displace [125I]hGH from monolayers by 35%. Adolescent cultured chondrocytes failed to demonstrate specific binding of [125I]hGH. We conclude that GH receptors are widely distributed in a range of mesenchyme cells in the human infant growth plate, including bone and hemopoietic precursors. The expression of these receptors appears to be maturation dependent in both intact tissue and culture, while they may no longer be expressed after the peak growth phase of puberty.

Demonstration and localization of growth hormone receptor in human skin and skin fibroblasts.

Clinical evidence suggests that skin is responsive to GH status in vivo. We sought to demonstrate the presence of GH receptors in human skin and in cultured skin fibroblasts using the techniques of immunohistochemistry and northern blotting. Human foreskin was obtained at surgery for preparation of sections and primary fibroblast cultures. Skin sections and fibroblast monolayers were immunostained using a monoclonal antibody which recognizes the hGH receptor (MAb 263). Positive immunoperoxidase staining was seen in all epidermal layers except the stratum corneum, in dermal sweat and sebaceous glands, and in dermal fibroblasts. In cultured fibroblasts capping of surface GH receptor was observed after aqueous formaldehyde fixation, whereas fixation in Carnoy's solution resulted in granular cytoplasmic staining. Fibroblast poly A+ RNA was prepared from cultured skin fibroblasts, separated by denaturing agarose gel electrophoresis, blotted onto nitrocellulose, and hybridized to a 32P-labeled, 847 base pair (bp) hGH receptor complementary DNA (cDNA) clone. Human liver and non-pregnant rabbit liver total RNA were used as controls. Fibroblast poly A+ RNA contained a single hybridizing species of approximately 5.2 kilobase. Human liver total RNA also contained a single hybridizing species of 4.9 kilobase. We have demonstrated the presence of GH receptor protein in human skin and growth hormone receptor mRNA and protein in cultured human skin fibroblasts. These observations suggest that GH may indeed have a direct role in modulating keratinocyte and fibroblast function.

Immunoreactive growth hormone receptor/binding protein is present on fibroblasts and in serum of Laron-type dwarfs.

Laron-type dwarfism is an autosomal recessive disorder characterised by extreme growth retardation and growth hormone (GH) resistance and has been shown in some cases to be associated with mutations in the GH receptor gene. Limited data suggest that in this condition specific liver GH binding is absent. In the majority of reported cases specific GH binding is also absent in serum. However it is not known whether the GH receptor and/or the serum GH binding protein are expressed in this condition. Using the techniques of immunohistochemistry and Northern blotting we have demonstrated that in cultured skin fibroblasts derived from four patients with Laron-type dwarfism the GH receptor gene is transcribed and the GH receptor protein is expressed on the cell surface. Further study of one of these patients, who has not previously been reported, has also revealed low but detectable levels of GH binding protein in serum using a two-site immunradiometric assay which does not depend on GH binding. These results indicate that the growth hormone receptor/binding protein is expressed in Laron-type dwarfism.

Autoradiographic localization of insulin-like growth factor I binding sites in the bovine adrenal gland and on cultured bovine adrenal chromaffin cells.

Abstract Previous studies have reported the presence of binding sites for insulin-like growth factor I (IGF-I) in membranes prepared from isolated bovine adrenal medullary cells, and IGF-I was found to regulate the secretory function of bovine chromaffin cells. In the present study, binding sites for IGF-I have been localized in sections of bovine adrenal gland and on cultured bovine adrenal medullary cells, using [(125) l][Thr(59)]-IGF-l as the ligand in conjunction with qualitative autoradiographic techniques. Binding sites were present throughout the adrenal gland and were distributed evenly over all cortical cell layers and over both adrenaline and noradrenaline cell types in the medulla. They were also present at lower density over blood vessels and nerve bundles and over the capsule. The binding of radioligand was to a single class of sites with K(d) 0.61 nM, and was completely displaced by excess unlabelled [Thr(59)]-IGF-l and by insulin (Actrapid, K(d) 1.04muM). Binding sites were also identified on single cells in primary monolayer cultures of bovine adrenal medullary cells. More than 96% of the cells possessed binding sites, although only 85% of such cells were chromaffin cells, as previously determined from dopamine beta-hydroxylase immunohistochemical staining. The results suggest that IGF-I may affect the maturation, growth or function not only of adrenal chromaffin cells but also of many others cell types in this tissue.

Consumer health information seeking on the Internet: the state of the art.

Increasingly, consumers engage in health information seeking via the Internet. Taking a communication perspective, this review argues why public health professionals should be concerned about the topic, considers potential benefits, synthesizes quality concerns, identifies criteria for evaluating online health information and critiques the literature. More than 70 000 websites disseminate health information; in excess of 50 million people seek health information online, with likely consequences for the health care system. The Internet offers widespread access to health information, and the advantages of interactivity, information tailoring and anonymity. However, access is inequitable and use is hindered further by navigational challenges due to numerous design features (e.g. disorganization, technical language and lack of permanence). Increasingly, critics question the quality of online health information; limited research indicates that much is inaccurate. Meager information-evaluation skills add to consumers' vulnerability, and reinforce the need for quality standards and widespread criteria for evaluating health information. Extant literature can be characterized as speculative, comprised of basic 'how to' presentations, with little empirical research. Future research needs to address the Internet as part of the larger health communication system and take advantage of incorporating extant communication concepts. Not only should research focus on the 'net-gap' and information quality, it also should address the inherently communicative and transactional quality of Internet use. Both interpersonal and mass communication concepts open avenues for investigation and understanding the influence of the Internet on health beliefs and behaviors, health care, medical outcomes, and the health care system.