==================================BSR04================================== 4. Effect of growth factors on bone growth or healing. Growth factors: in general or, Epidermal Growth Factor (EGF); Transforming Growth Factor (TGF-beta/alpha); Platelet-derived Growth Factor (PDGF). Bone; Bone formation; Bone induction. Bone healing, Bone Resorbtion. 1 UI - 87126974 AU - McGraw ME ; Price DA ; Hill DJ TI - Somatomedin C deficiency in Asian sisters. AB - Two sisters of Asian origin showed typical clinical and biochemical features of primary somatomedin C (SM-C) deficiency (Laron dwarfism). Abnormalities of SM-C binding proteins were observed, one sister lacking the high molecular weight (150 Kd) protein. MH - Carrier Proteins/BLOOD ; Case Report ; Child, Preschool ; Dwarfism/BLOOD/ *FAMILIAL & GENETIC ; Female ; Human ; Infant ; Insulin-Like Growth Factor I/*DEFICIENCY ; Somatomedins/*DEFICIENCY SO - Arch Dis Child 1986 Dec;61(12):1233-5 2 UI - 87111934 AU - Hanks CT ; Kim JS ; Edwards CA TI - Growth control of cultured rat calvarium cells by platelet-derived growth factor. AB - Platelet-derived growth factor (PDGF), which is released during the "clotting cascade:, has a mitogenic effect on a variety of mesenchymal cells including those of the periosteum. Cell culture provides a method of studying these effects. In the present study, PDGF was studied for its ability to stimulate quiescent calvarium periosteal cells to enter G1, indicated by new protein synthesis. After addition of platelet poor plasma, these cells later entered S phase, indicated by uptake of 3H-thymidine. Three hours after PDGF stimulation, there were quantitative increases of at least 18 protein bands as compared to cells left in serum starvation medium. Two of these new protein bands (31K and 45K) required transcription. These data suggest that PDGF in extravasated blood may stimulate appositional bone formation beneath intact periosteum at the site of injury. MH - Actinomycin/PHARMACODYNAMICS ; Animal ; Cell Division/*DRUG EFFECTS ; Cells, Cultured ; Cycloheximide/PHARMACODYNAMICS ; DNA/BIOSYNTHESIS ; Fetus ; Periosteum/CYTOLOGY ; Platelet-Derived Growth Factor/ *PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Skull/*CYTOLOGY SO - J Oral Pathol 1986 Oct;15(9):476-83 3 UI - 87111655 AU - Cie:slak D ; Szulc-Kuberska J ; Stepie:n H ; Klimek A TI - Epidermal growth factor in human cerebrospinal fluid: reduced levels in amyotrophic lateral sclerosis. AB - Epidermal growth factor (EGF), a mitogenic peptide, is widely distributed within the brain and endocrine cells of the gastro-intestinal tract. Using EGF radioreceptor assay, the EGF level was measured in lumbar cerebrospinal fluid from five patients with amyotrophic lateral sclerosis (ALS) and seven patients with intervertebral disc disease as a control group. The patients with ALS showed reduced EGF levels to 662.4 +/- 207 pg/ml as compared with controls 1013 +/- 182.8 pg/ml (P less than 0.02). These results indicate a possible EGF involvement in the pathogenesis of ALS. MH - Aged ; Amyotrophic Lateral Sclerosis/*CEREBROSPINAL FLUID ; Comparative Study ; Epidermal Growth Factor-Urogastrone/*CEREBROSPINAL FLUID ; Human ; Intervertebral Disk ; Middle Age ; Spinal Diseases/CEREBROSPINAL FLUID ; Support, Non-U.S. Gov't SO - J Neurol 1986 Nov;233(6):376-7 4 UI - 87100644 AU - Linkhart TA ; Jennings JC ; Mohan S ; Wakley GK ; Baylink DJ TI - Characterization of mitogenic activities extracted from bovine bone matrix. AB - The mitogenic activity in the unfractionated mixture of proteins released from adult bovine bone matrix during demineralization with ethylenediaminetetraacetate (EDTA) has been examined. Bovine bone extract (BBE) from 1 to 25 micrograms protein per ml stimulated proliferation of chick embryo calvaria bone cells, newborn mouse bone cells, and osteoblastlike cell lines MMB-1 and ROS 17/2.8. BBE also stimulated DNA synthesis in cells from chick embryo cartilage, skin and skeletal muscle tissues and fibroblastlike BALB/c 3T3 and NRK cells. BBE contained beta transforming growth factor (TGF) activity (NRK cell colony formation in soft agar in the presence of epidermal growth factor EGF). The cell specificity results suggest that BBE contains more than one growth factor, including a beta TGF and a factor that is not specific for bone cells, and all of the bone derived growth factor activities that have been described previously, including SGF, are apparently present in BBE. Maximal stimulation of chick embryo calvarial cell DNA synthesis by BBE was equal to or exceeded maximal stimulation by nonosseous growth factors that have been reported to stimulate DNA synthesis in bone organ cultures (EGF, fibroblast growth factor, platelet-derived growth factor, insulinlike growth factor I, and multiplication stimulating activity). Combinations of BBE with maximally stimulatory concentrations of each growth factor stimulated DNA synthesis to a greater magnitude than did each growth factor alone. These results suggest that combinations of bone derived and systemic factors can coordinately stimulate bone cell proliferation. MH - Animal ; Bone and Bones/CYTOLOGY/METABOLISM ; Bone Matrix/*ANALYSIS ; Cattle ; Cell Division ; Cell Line ; Chick Embryo ; Comparative Study ; DNA/BIOSYNTHESIS ; Growth Substances/*PHARMACODYNAMICS ; Mice ; Organ Culture ; Osteoblasts/CYTOLOGY ; Peptides/PHARMACODYNAMICS ; Rats ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; Tissue Extracts/PHARMACODYNAMICS SO - Bone 1986;7(6):479-87 5 UI - 87074913 AU - Komoda T ; Koyama I ; Nagata A ; Sakagishi Y ; Kurata M ; Kumegawa M TI - A possible mechanism of induction and translocation into blood stream of rat alkaline phosphatase activity by bile duct ligation. AB - We investigated the effects of bile duct ligation on alkaline phosphatase (ALP) activities in liver, calvarium, duodenum, and ileum in rats and its possible mechanism of action. ALP isozyme activities in the ligated rats were significantly elevated in the liver and duodenum, while those in the ileum and calvarium were markedly decreased. The ALP isozyme activity elevated by the ligation was obviously suppressed by prior administration of indomethacin, an inhibitor of prostaglandin synthesis. Moreover, phorbol ester also elevated the ALP activity as well as the phosphatase level in the ligated rat. However, other drugs, such as an inhibitor of protein kinase C and calmodulin, showed different effects: calmodulin stimulated an 11.0-, 1.3-, or 1.5-fold increase in ALP activity in the ileum, duodenum, or calvarium, respectively; whereas the hepatic enzyme activity was not affected. The induction by calmodulin was markedly different from that by the ligation. Moreover, imipramine, an inhibitor of protein kinase C, had little effect. These results suggest that prostaglandin is a possible ALP inducer in ligated rats, probably working by elevating the cAMP level. On the other hand, the ligation induced simultaneously de novo synthesis of the membranous and soluble ALP isozymes; and the release rate of the soluble enzyme was greater than that of the membranous isozymes, indicating that the soluble enzyme might be a main source of the induced serum ALP. Lectin affinity chromatography indicated that the soluble enzyme or induced serum enzyme may contain more fucose than that of the membranous one, suggesting that the sugar moiety in the ALP molecule may relate to the clearance of ALP from or its release into the circulation. MH - Adenosine Cyclic Monophosphate/METABOLISM ; Alkaline Phosphatase/BLOOD/ *METABOLISM ; Animal ; Bile Ducts/*PHYSIOLOGY/SURGERY ; Bone and Bones/ ENZYMOLOGY ; Calmodulin/METABOLISM ; Chlorpromazine/PHARMACODYNAMICS ; Cholestasis/*ENZYMOLOGY ; Enzyme Induction ; Epidermal Growth Factor-Urogastrone/METABOLISM ; Fucose/METABOLISM ; Indomethacin/ PHARMACODYNAMICS ; Intestines/ENZYMOLOGY ; Isoenzymes/METABOLISM ; Ligation ; Liver/ENZYMOLOGY ; Prostaglandins/METABOLISM ; Protein Kinase C/METABOLISM ; Rats ; Tetradecanoylphorbol Acetate/PHARMACODYNAMICS SO - Arch Biochem Biophys 1986 Nov 15;251(1):323-35 6 UI - 87044839 AU - Partsch CJ ; Hermanussen M ; Sippell WG TI - Treatment of Silver-Russell type dwarfism with human growth hormone: effects on serum somatomedin-C levels and on longitudinal growth studied by knemometry. AB - Two patients, aged 5 and 12 years, with Silver-Russell type dwarfism are presented. As shown by standard tests and examination of the spontaneous nocturnal hGH secretion, there was only mild regulative hGH deficiency. HGH treatment was started with daily subcutaneous injections at age 5.5 years (height 90.3 cm, -6.1 SDS, bone age 2.75 years) and 12.6 years (height 125.7 cm, -3.7 SDS, bone age 8.75 years), respectively. Treatment was monitored by serial somatomedin-C (SM-C) determinations and by knemometry (lower leg measurement). SM-C values increased in both patients by 10.5 and 4.8 fold, respectively, and remained above the prepubertal range (greater than 2.5 U/ml) during the treatment periods of 1.5 years. Pretreatment knemometric growth rate was high (after a somatomedin generation test) in patient 1 (0.7 mm/week) and low in patient 2 (0.31 mm/week). It remained at the same level in patient 1 (0.67 mm/week) and increased markedly in patient 2 (0.46 mm/week). During a treatment interruption, in both patients, knemometric growth rates fell to 0.33 and 0.30 mm/week, respectively. After resumption of treatment, now with biosynthetic hGH, growth rates increased again in patients 1 and 2 to 0.64 and 0.48 mm/week, respectively. This lower leg growth pattern was parallelled by similar changes in total body growth velocity. Even after the relatively short treatment period of 14 to 16 months, a slight net gain in statural height could be observed, as standard deviation scores for bone age increased. MH - Age Determination by Skeleton ; Anthropometry ; Body Height ; Case Report ; Child ; Child, Preschool ; Dwarfism/*DRUG THERAPY/PHYSIOPATHOLOGY ; Human ; Insulin-Like Growth Factor I/*BLOOD ; Leg ; Male ; Somatomedins/ *BLOOD ; Somatotropin/*THERAPEUTIC USE SO - Acta Endocrinol [Suppl] (Copenh) 1986;279:139-46 7 UI - 87033424 AU - Hale LV ; Hale JE ; Kemick ML ; Ishikawa Y ; Wuthier RE TI - Development of a new serum-free medium, USC-HC1, for growth and normal phenotype in postembryonic chicken growth plate chondrocytes. AB - A serum-free medium for postembryonic chicken epiphyseal growth plate chondrocytes has been developed from 104 MCDB medium. To enable these fastidious cells to survive, grow, and express normal phenotype, a substantial increase over MCDB 104 in the level of many of the amino acids was required, as well as a change in the buffer system and the addition of SerXtend, a defined, serum-free product containing various growth factors, including fibroblast growth factor. Also required was the provision of cell attachment factors, either by coating culture surfaces with type II collagen, or better, by allowing the freshly released cells to recover for several hours in a medium supplemented with 10% fetal bovine serum before plating. This new serum-free medium, which we call USC-HC1, supports growth and replication, the retention of normal polygonal morphology, the expression of significant levels of cellular alkaline phosphatase activity, the production of sulfated proteoglycans, type II collagen, and the formation of alkaline phosphatase-rich matrix vesicles by the chondrocytes. The major advantage of USC-HC1, however, is that it will provide for the first time an opportunity to examine the effects of various defined growth and hormonal factors on the phenotypic expression and differentiation of growth plate chondrocytes, in the absence of the variable (stimulatory and inhibitory) factors present in fetal bovine serum. MH - Alkaline Phosphatase/ANALYSIS ; Animal ; Cell Differentiation ; Cell Division ; Cells, Cultured ; Chickens ; *Culture Media ; Extracellular Matrix/METABOLISM ; Growth Plate/*CYTOLOGY/GROWTH & DEVELOPMENT/ METABOLISM ; Growth Substances/PHARMACODYNAMICS ; Phenotype ; Support, U.S. Gov't, P.H.S. SO - In Vitro Cell Dev Biol 1986 Oct;22(10):597-603 8 UI - 86318937 AU - Thorner MO ; Vance ML ; Evans WS ; Ho K ; Rogol AD ; Blizzard RM ; Furlanetto R ; Rivier J ; Vale W TI - Growth hormone releasing factor and somatomedin C production: extrahypothalamic localization and possible functional significance. AB - Growth hormone-releasing factor (GRF) is found in the highest concentration (albeit lower compared to other hypothalamic regulatory hormones) in the hypothalamus. There is mounting evidence that GRF-like immunoreactivity is found in other sites in the CNS and in the periphery. The role of GRF, other than to stimulate growth hormone secretion by the somatotroph, is unknown. In addition generation of IGF-1 in response to GRF appears to be dependent on an intact pituitary. MH - Animal ; Endocrine Diseases/PHYSIOPATHOLOGY ; Growth Plate/DRUG EFFECTS ; Human ; Hypothalamus/PHYSIOLOGY ; Insulin-Like Growth Factor I/PHYSIOLOGY/ *SECRETION ; Intestines/PHYSIOLOGY ; Male ; Neoplasms/PHYSIOPATHOLOGY ; Pituitary Gland, Anterior/PHYSIOLOGY ; Rats ; Somatomedins/*SECRETION ; Somatotropin/DEFICIENCY/SECRETION ; Somatotropin Releasing Hormone/ PHARMACODYNAMICS/PHYSIOLOGY/*SECRETION SO - Acta Endocrinol [Suppl] (Copenh) 1986;276:34-40 9 UI - 86312621 AU - van Buul-Offers S ; Ueda I ; Van den Brande JL TI - Biosynthetic somatomedin C (SM-C/IGF-I) increases the length and weight of Snell dwarf mice. AB - An Escherichia coli derived somatomedin-C/IGF-I preparation (rec-IGF-I) with an amino acid sequence identical to the natural IGF-I derived from human plasma, increases body length and weight, as well as the growth of several organs of Snell dwarf mice, when administered for 4 wk. After 2 wk of treatment rec-IGF-I (22.2 micrograms/day) induced a significant increase over buffer treated controls, to a comparable degree as obtained with bacterially synthesized human growth hormone (bhGH; 8.4 micrograms/day). The weight/length ratio of rec-IGF-I and bhGH-treated dwarf mice after 4 wk of treatment were not significantly different. A significant increase over controls was obtained with both preparations. Organs with increased weights after bhGH treatment (brain; submandibular salivary glands; heart, liver, kidneys, thymus, and spleen) were also heavier after rec-IGF-I. Significance was only reached for the kidneys and the spleen and the musculus quadriceps femoris. Organ weights expressed as a percentage of body weight of bhGH and rec-IGF-I treated dwarfs were similar except for the relative weight of the heart of the bhGH group, which was significantly increased compared to the controls and the rec-IGF-I group. These data resolve the issue as to whether or not pure SM-C/IGF-I will induce growth in length and demonstrate the usefulness of recombinant IGF-I in the studies of growth regulation. MH - Animal ; Biometry ; Body Weight/*DRUG EFFECTS ; Dwarfism/PHYSIOPATHOLOGY ; Human ; Insulin-Like Growth Factor I/*PHARMACODYNAMICS ; Mice ; Mice, Mutant Strains/*ANATOMY & HISTOLOGY ; Organ Weight/DRUG EFFECTS ; Recombinant Proteins/*PHARMACODYNAMICS ; Somatomedins/*PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Time Factors SO - Pediatr Res 1986 Sep;20(9):825-7 10 UI - 86304448 AU - Hauschka PV ; Mavrakos AE ; Iafrati MD ; Doleman SE ; Klagsbrun M TI - Growth factors in bone matrix. Isolation of multiple types by affinity chromatography on heparin-Sepharose. AB - The mineralized matrix of osseous tissue harbors abundant mitogenic activity which is extractable by demineralizing solvents. In bovine bone powder free of blood and cartilage contamination, the volume concentration of mitogens is up to 20 times greater than in serum. Growth factor activity in bone extracts was quantitated on quiescent mouse BALB/c/3T3 fibroblasts, where [3H]thymidine incorporation for 48 h was stimulated up to 200-fold in a linear, dose-dependent manner. Six distinct bone-derived growth factors (BDGFs) have been resolved and partially purified (up to 44,000-fold) on heparin-Sepharose using NaCl gradient elution. Provisionally named by the NaCl molarity at which they elute, these BDGFs include BDGF-0.45 (25% of total activity). This factor is heat-stable and sensitive to dithiothreitol, and displaces 125I-labelled bovine platelet-derived growth factor in a radioreceptor assay. BDGF-0.45 (approximately 50 ng/g of bone) is closely related or identical to bovine platelet-derived growth factor. BDGF-1.1 (10%) has a pI of 5.2 and shows a 16,600-dalton doublet on sodium dodecyl sulfate-polyacrylamide gel electrophoresis Western blots stained with antiserum to bovine anionic fibroblast growth factor. Two activities with high heparin affinity resemble cationic forms of fibroblast growth factor. BDGF-1.5 is the dominant factor in fetal membranous bone (50%), but is less abundant in adult bone (20%). BDGF-1.7, a 17,500-18,400-dalton triplet, is virtually absent in fetal bone (7%) but abundant (30%) in adult bone and may be related to cartilage derived growth factor. Two minor activities, BDGF-0.1 (10%) and BDGF-2.0 (7%) have not been characterized. Proliferation of bovine capillary endothelial cells was strongly supported by BDGFs 1.1, 1.5, and 1.7, but not by 0.45. These four purified BDGFs and the crude bone extract were also strongly mitogenic for rat osteoblasts while depressing alkaline phosphatase specific activity by 2-3-fold. Bone exhibits the most complex spectrum of growth factor activities of any tissue yet described. Bone cells and other indigenous cell types must be considered as possible sources of the BDGFs, in addition to sequestration from blood. Mechanisms for unmasking or release of BDGFs from the mineralized matrix resulting in local action on target cells are undoubtedly important for the development and maintenance of bone tissue. MH - Age Factors ; Animal ; Biological Assay ; Bone Matrix/*ANALYSIS ; Cattle ; Cell Division/DRUG EFFECTS ; Chromatography, Affinity ; Fibroblasts/ DRUG EFFECTS ; Growth Substances/*ISOLATION & PURIFICATION ; Isoelectric Focusing ; Mice ; Mice, Inbred BALB C ; Sepharose/ANALOGS & DERIVATIVES ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Sep 25;261(27):12665-74 11 UI - 86300772 AU - Easty DM ; Easty GC ; Baici A ; Carter RL ; Cederholm-Williams SA ; Felix H ; Gusterson B ; Haemmerli G ; Hauser-Urfer I ; Heizmann CW ; et al TI - Biological studies of ten human squamous carcinoma cell lines: an overview. AB - Ten cell lines established from surgical specimens of human squamous carcinomas of the tongue and larynx have been investigated with respect to their motility, ultrastructure, karyotypes, certain biochemical features, interaction with normal epithelial and stromal elements and capacity to infiltrate three-dimensional organoid systems. All the cell lines have maintained several morphological and biochemical characteristics indicating a common origin, although the extent to which each line displays this heritage is variable. The phenotypes of each of the individual cell lines are, however, notably stable. Data are provided for epithelial surface markers (including epidermal growth factor, EGF) and for the synthesis and release of prostaglandins and proteases which may be involved in invasive mechanisms. Encounters between the cell lines and organoid substrata (embryonic chick heart spheroids, human amnion, chick chorioallantoic membrane) are described: the results indicate a scale of invasiveness ranging from lack of penetration to full-thickness infiltration by cells showing various distinctive growth patterns. Correlation between in vitro and in vivo findings is discussed, and it is suggested that the biological heterogeneity of the lines may reflect inherent properties of the original carcinoma cell populations which are more distinctly expressed in vitro. MH - Aged ; Animal ; Bone and Bones/PHYSIOPATHOLOGY ; Calcium-Binding Proteins/ METABOLISM ; Carcinoembryonic Antigen/METABOLISM ; Carcinoma, Squamous Cell/METABOLISM/*ULTRASTRUCTURE ; Cartilage/PHYSIOPATHOLOGY ; Cell Communication ; Cell Line ; Cell Movement ; Chick Embryo ; Epidermal Growth Factor-Urogastrone/METABOLISM ; Female ; Human ; Karyotyping ; Keratin/METABOLISM ; Laryngeal Neoplasms/ULTRASTRUCTURE ; Male ; Membrane Proteins/METABOLISM ; Microscopy, Electron ; Middle Age ; Review ; Support, Non-U.S. Gov't ; Tongue Neoplasms/ULTRASTRUCTURE SO - Eur J Cancer Clin Oncol 1986 Jun;22(6):617-34 12 UI - 86278282 AU - Trippel SB ; Van Wyk JJ ; Mankin HJ TI - Localization of somatomedin-C binding to bovine growth-plate chondrocytes in situ. AB - The somatomedins are a family of low-molecular-weight peptides that are thought to mediate the stimulatory effect of growth hormone on skeletal growth. The cells that are directly responsible for skeletal growth are the chondrocytes of the epiphyseal growth plate, and these are the presumed skeletal target cells for somatomedin. As with other peptide growth factors, the cellular effects of the somatomedins are initiated by the interaction of the growth factor with specific receptors on target-cell surface membranes. Chondrocytes that have been isolated from bovine growth plates were previously found to possess specific surface receptors for the principal growth-hormone-dependent somatomedin, somatomedin-C. These studies indicated that the interaction of growth-plate chondrocytes with somatomedin-C involves specific receptor-binding followed by somatomedin-C internalization by the cell, a process identical to that identified in the mechanism of action of other peptide growth factors in other cells. These studies, however, left unanswered the questions of whether there are differences in binding of somatomedin-C by the different cell populations within the physis and whether somatomedin-C has access to cells in intact tissues. The current studies address these issues and indicate that bovine physeal chondrocytes in situ are accessible to exogenous somatomedin-C, that they specifically bind somatomedin-C in situ, and that cells of different physeal zones bind somatomedin-C differently. Labeled somatomedin-C is specifically bound by cells of all physeal zones. However, the binding is greatest for those cells undergoing active synthesis of DNA in the proliferative zone.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Animal ; Autoradiography ; Binding Sites ; Bone Development ; Cartilage/ CYTOLOGY/METABOLISM ; Cattle ; Cell Division ; Growth Plate/CYTOLOGY/ *METABOLISM ; Human ; Insulin-Like Growth Factor I/*METABOLISM ; Iodine Radioisotopes/DIAGNOSTIC USE ; Receptors, Endogenous Substances/ METABOLISM ; Somatomedins/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Thymidine/METABOLISM ; Tritium/DIAGNOSTIC USE SO - J Bone Joint Surg [Am] 1986 Jul;68(6):897-903 13 UI - 86274997 AU - Hizuka N ; Takano K ; Shizume K ; Asakawa K ; Miyakawa M ; Tanaka I ; Horikawa R TI - Insulin-like growth factor I stimulates growth in normal growing rats. AB - The scarcity of purified somatomedin/insulin-like growth factor (SM/IGF) has prevented investigation of the mechanisms of SM/IGF action. Recently insulin-like growth factor I (IGF-I) has been synthesized by recombinant DNA technology. The availability of large quantities of the biosynthetic IGF-I made it possible to study its effects by administering 120 micrograms/day via s.c. implanted minipump to rats for 7 days. After this 7 day administration of IGF-I, the body weight increased to 197.6 +/- 3.5% of initial values; the value was significantly greater than that of the control (179.4 +/- 3.7% of initial values, P less than 0.01). The body length and tibial epiphyseal width in IGF-I-treated rats were also significantly increased over those of control rats. The weights of kidneys, liver, testes and pituitary in IGF-I-treated rats were greater than those in control rats as well. These results provide a first demonstration that IGF-I stimulates growth in normal rats in vivo, and suggest that IGF-I might be useful in the treatment of growth retardation. MH - Animal ; Body Weight/DRUG EFFECTS ; Growth/*DRUG EFFECTS ; Growth Plate/ DRUG EFFECTS ; Insulin-Like Growth Factor I/BLOOD/*PHARMACODYNAMICS ; Male ; Organ Weight/DRUG EFFECTS ; Rats ; Rats, Inbred Strains ; Somatomedins/*PHARMACODYNAMICS ; Support, Non-U.S. Gov't SO - Eur J Pharmacol 1986 Jun 5;125(1):143-6 14 UI - 86272877 AU - Burwell RG ; Vernon CL ; Dangerfield PH ; Hall DJ ; Kristmundsdottir F TI - Raised somatomedin activity in the serum of young boys with Perthes' disease revealed by bioassay. A disease of growth transition? AB - This paper reports a study of the serum somatomedin activity in 67 boys with Perthes' disease and in 43 control boys aged three to 11 years. It was undertaken to evaluate some abnormalities of growth in children with Perthes' disease that have been previously reported. A brief account is given of knowledge relating to somatomedins in postnatal and fetal life. Serum somatomedin activity was measured using a bioassay based on the principle that somatomedins stimulate the synthesis of both DNA and proteoglycans in porcine costal cartilage. In control boys, the serum somatomedin activity increased with age, which is consistent with previous reports for normal children. In affected boys, the normal increase in serum somatomedin activity with age did not occur. The somatomedin activity in affected boys is higher than in control boys at three to five years but not at six to 11 years of age. Findings support the hypothesis that some children with Perthes' disease have an abnormality of the growth hormone-dependent somatomedins. The serum findings together with those of both skeletal age delay and impaired skeletal growth distally in the limbs are consistent with the view that the general disorder of some children with Perthes' disease results from an imbalance in mechanisms that determine the postnatal transition from the "fetal: to the "basic: component of the normal human growth curve. MH - Bone Diseases, Developmental/BLOOD/METABOLISM ; Child ; Child, Preschool ; Femur Head Necrosis/*BLOOD ; Gigantism/BLOOD ; Human ; Hydrocortisone/ BLOOD ; Legg-Perthes Disease/*BLOOD/METABOLISM ; Male ; Somatomedins/ *BLOOD/METABOLISM ; Support, Non-U.S. Gov't SO - Clin Orthop 1986 Aug;(209):129-38 15 UI - 86272813 AU - Bijlsma JW ; Duursma SA TI - Serum concentrations of somatomedins and growth hormone in relation to bone metabolism in acromegaly and thyroid dysfunction. AB - Many hormones are involved in the complex process of formation and resorption of bone. However, only somatomedin is found to directly stimulate cell replication and collagen synthesis in bone. This study was undertaken to examine a possible regulatory role of somatomedin in mediating the effects of growth hormone and thyroid hormones on bone metabolism. Bone metabolism and concentrations of somatomedins and growth hormones were studied in 17 acromegalic, 15 thyrotoxic and 14 hypothyroid patients, before and during treatment. During treatment of acromegalic and thyrotoxic patients parameters of bone turnover, both formation and resorption, decreased parallel to the decrease in concentration of somatomedin. During treatment of hypothyroid patients parameters of bone turnover increased. A positive correlation was found in acromegalic patients between changes in somatomedins and parameters of bone resorption (R = 0.82, P less than 0.01) as well as bone formation (R = 0.63, P less than 0.05) and in thyrotoxic patients between changes in somatomedin and bone resorption (R = 0.87, P less than 0.05). These data suggest that somatomedin may indeed play a role in the regulation of bone turnover. In addition, secondary effects on growth hormone concentrations were observed. MH - Acromegaly/*BLOOD/METABOLISM ; Bone and Bones/*METABOLISM ; Female ; Human ; Hyperthyroidism/BLOOD/METABOLISM ; Hypothyroidism/BLOOD/ METABOLISM ; Male ; Middle Age ; Osmolar Concentration ; Somatomedins/ *BLOOD ; Somatotropin/*BLOOD ; Thyroid Diseases/*BLOOD/METABOLISM SO - Clin Exp Rheumatol 1986 Apr-Jun;4(2):105-10 16 UI - 86261814 AU - Nilsson A ; Isgaard J ; Lindahl A ; Dahlstr:om A ; Skottner A ; Isaksson OG TI - Regulation by growth hormone of number of chondrocytes containing IGF-I in rat growth plate. AB - Whether growth hormone stimulates longitudinal bone growth by a direct effect at the site of the growth plate or indirectly by increasing the concentration of circulating somatomedins (insulin-like growth factors) has been the subject of controversy. Immunohistochemical methods were used to explore the localization and distribution of insulin-like growth factor I (IGF-I) immunoreactivity in the epiphyseal growth plate of the proximal tibia of male rats. Cells in the proliferative zone of the growth plate of normal rats exhibited a bright immunofluorescence, whereas cells in the germinal and hypertrophic zones stained only weakly. In rats subjected to hypophysectomy, the number of fluorescent cells was markedly reduced. When the hypophysectomized rats were treated with growth hormone, either systemically or at the site of the growth plate, the number of IGF-I-immunoreactive cells in the proliferative zone was increased. The results show that IGF-I is produced in proliferative chondrocytes in the growth plate and that the number of IGF-I-containing cells is directly regulated by growth hormone. These findings suggest that IGF-I has a specific role in the clonal expansion of differentiated chondrocytes and exerts its function locally through autocrine or paracrine mechanisms. MH - Animal ; Fluorescent Antibody Technic ; Growth Plate/*CYTOLOGY/DRUG EFFECTS/GROWTH & DEVELOPMENT ; Hypophysectomy ; Insulin-Like Growth Factor I/PHARMACODYNAMICS/*PHYSIOLOGY ; Male ; Rats ; Rats, Inbred Strains ; Somatomedins/*PHYSIOLOGY ; Somatotropin/PHARMACODYNAMICS/ *PHYSIOLOGY ; Support, Non-U.S. Gov't ; Tibia SO - Science 1986 Aug 1;233(4763):571-4 17 UI - 86251292 AU - Sato K ; Mimura H ; Han DC ; Kakiuchi T ; Ueyama Y ; Ohkawa H ; Okabe T ; Kondo Y ; Ohsawa N ; Tsushima T ; et al TI - Production of bone-resorbing activity and colony-stimulating activity in vivo and in vitro by a human squamous cell carcinoma associated with hypercalcemia and leukocytosis. AB - A squamous cell carcinoma of 33-yr-old patient who developed marked leukocytosis and hypercalcemia was transplanted into nude mice in which more marked leukocytosis and hypercalcemia also developed. This tumor (LJC-1-JCK) produced a colony-stimulating factor (CSF) and formed a cyst in the tumor from which a CSF-producing cell line (T3M-1) was established. The CSF causes predominantly formation of granulocytic colonies in addition to macrophage colonies. Bone-resorbing activity (BRA) was detected in the cystic fluid and was eluted as two separate peaks with proteins of an apparent molecular weight of 30,000-50,000 and 10,000-20,000. Colony-stimulating activity (CSA) was eluted at an apparent 30,000 mol wt. The conditioned medium of the T3M-1 cells also contained a BRA with an apparent 14,000 mol wt, whereas CSA eluted at an apparent 30,000 mol wt. PTH, epidermal growth factor, transforming growth factor-alpha, prostaglandin Es, and vitamin D could not account for the powerful BRA. In contrast to CSA, BRA was not inactivated by trypsin and more stable at 70 degrees C. When T3M-1 cells were transplanted into nude mice, marked hypercalcemia developed in addition to granulocytosis. Our findings suggest that the tumor produces and secretes a powerful BRA in vivo and in vitro, which is different from CSA in terms of molecular weight, heat stability, and trypsin treatment. We speculate that the synergistic action of CSF that stimulates macrophage colony formation and recruits osteoclast precursors, and BRA, which stimulates mononuclear phagocytes and/or osteoclasts were responsible for a marked increase in osteoclastic bone resorption and humoral hypercalcemia in the patient. MH - Animal ; *Bone Resorption ; Carcinoma, Squamous Cell/COMPLICATIONS/ *METABOLISM ; Chromatography, Gel ; Colony-Stimulating Factor/*METABOLISM ; Culture Media ; Epidermal Growth Factor-Urogastrone/ANALYSIS ; Exudates and Transudates/ANALYSIS ; Heat ; Human ; Hydrocortisone/PHARMACODYNAMICS ; Hypercalcemia/*COMPLICATIONS ; Indomethacin/PHARMACODYNAMICS ; Interleukin 1/ANALYSIS ; Leukocytosis/*COMPLICATIONS ; Mice ; Molecular Weight ; Parathyroid Hormones/ANALYSIS ; Prostaglandins/ANALYSIS ; Prostaglandins E/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Trypsin/METABOLISM ; Vitamin D/METABOLISM SO - J Clin Invest 1986 Jul;78(1):145-54 18 UI - 86224651 AU - Lorenzo JA ; Quinton J ; Sousa S ; Raisz LG TI - Effects of DNA and prostaglandin synthesis inhibitors on the stimulation of bone resorption by epidermal growth factor in fetal rat long-bone cultures. AB - We examined two inhibitors of DNA synthesis, hydroxyurea (HU) and aphidicholin (APC), and two inhibitors of prostaglandin cyclooxygenase, indomethacin and flufenamic acid, for their effects on the resorptive responses of fetal rat long-bone cultures to epidermal growth factor (EGF) and parathyroid hormone (PTH). As we have previously found, HU decreased unstimulated 45Ca release but had little effect on the resorptive response to PTH. HU also did not block resorption stimulated by EGF. Addition of the cyclooxygenase inhibitor, indomethacin, did not alter the resorptive responses of unstimulated or PTH-treated cultures in either the presence or absence of HU or the resorptive response of bones cultured with EGF alone. However, indomethacin completely blocked the resorptive response to EGF of bones that were cultured with HU. The effects of indomethacin on EGF-mediated resorption in HU-treated cultures appeared to be related to an inhibition of prostaglandin synthesis since flufenamic acid had similar effects. However, the effects of HU on the resorptive response to EGF may not have resulted solely from its inhibitory action on DNA synthesis since APC, in the absence of cyclooxygenase inhibitors, completely blocked EGF-mediated resorption without significantly affecting the response to PTH. These results demonstrate that the mechanisms regulating PTH- and EGF-mediated resorption in fetal rat long-bone cultures differ, and imply that a component of EGF-mediated resorption in these cultures is dependent on sustained DNA synthesis. MH - Animal ; Bone Resorption/*DRUG EFFECTS ; Diterpenes/PHARMACODYNAMICS ; DNA Replication/*DRUG EFFECTS ; Epidermal Growth Factor-Urogastrone/ *PHARMACODYNAMICS ; Female ; Hydroxyurea/PHARMACODYNAMICS ; Indomethacin/ PHARMACODYNAMICS ; Organ Culture ; Parathyroid Hormones/PHARMACODYNAMICS ; Pregnancy ; Prostaglandins/*BIOSYNTHESIS ; Rats ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; Thymidine/METABOLISM SO - J Clin Invest 1986 Jun;77(6):1897-902 19 UI - 86214656 AU - Kukita T ; Kurisu K TI - Interaction between epidermal growth factor and triamcinolone acetonide in mouse palatal mesenchymal cells in vitro. AB - Palatal mesenchymal cells from mouse embryos (MEPM cells) had a high number of specific receptors for epidermal growth factor (EGF). At 37 degrees C, the number of high-affinity receptors (dissociation constant, Kd of 6.0 X 10(-10) M) was 2.0 X 10(5) per cell and of low-affinity receptors (Kd of 2.9 X 10(-9) M), 3.8 X 10(5) per cell. At 2 degrees C, a single class of receptors (Kd of 4.8 X 10(-9) M) was detected at 1.8 X 10(5) per cell. Triamcinolone acetonide, a potent cleft-palate-inducing agent, slightly inhibited the recovery of 125I-labelled EGF binding capacity in MEPM cells after down regulation of EGF receptors; it did not affect the binding properties of 125I-labelled EGF in these cells. MH - Animal ; Drug Interactions ; DNA/BIOSYNTHESIS ; Epidermal Growth Factor-Urogastrone/*METABOLISM/PHARMACODYNAMICS ; In Vitro ; Male ; Mice ; Mice, Inbred Strains ; Palate/EMBRYOLOGY/*METABOLISM ; Protein Binding/ DRUG EFFECTS ; Receptors, Endogenous Substances/BIOSYNTHESIS ; Triamcinolone Acetonide/*METABOLISM/PHARMACODYNAMICS SO - Arch Oral Biol 1986;31(1):39-44 20 UI - 86196719 AU - Yoneda T ; Urade M ; Sakuda M ; Miyazaki T TI - Altered growth, differentiation, and responsiveness to epidermal growth factor of human embryonic mesenchymal cells of palate by persistent rubella virus infection. AB - We previously demonstrated that human embryonic mesenchymal cells derived from the palate (HEMP cells) retain alkaline phosphatase (ALP) content and capacity for collagen synthesis after long-term culture, and their growth is markedly stimulated by epidermal growth factor (EGF). There was a dramatic decrease in ALP content and capacity to synthesize collagen in HEMP cells (HEMP-RV cells) persistently infected with rubella virus (RV). EGF increased ALP activity and decreased collagen synthesis in HEMP cells, whereas EGF showed no effect on these activities in HEMP-RV cells. Growth of HEMP-RV cells was slightly reduced compared with that of HEMP cells. EGF stimulated growth of HEMP cells and to a lesser extent of HEMP-RV cells. Binding of 125I-EGF to cell-surface receptors in HEMP-RV cells was, to our surprise, twice as much as that in HEMP cells. However, internalization of bound 125I-EGF in HEMP-RV cells was profoundly diminished. Thus, persistent RV infection causes not only changes in HEMP cell growth and differentiation but a decrease in or loss of HEMP cell responsiveness to EGF. The effects of persistent RV infection on palatal cell differentiation as well as growth may be responsible for the pathogenesis of congenital rubella. Furthermore, since HEMP cells appear to be closely related to osteoblasts, these results suggest a mechanism for RV-induced osseous abnormalities manifested in congenital rubella patients. MH - Alkaline Phosphatase/ANALYSIS ; Bone Development ; Cell Differentiation/ DRUG EFFECTS ; Cell Division/DRUG EFFECTS ; Cells, Cultured ; Collagen/ BIOSYNTHESIS ; Epidermal Growth Factor-Urogastrone/METABOLISM/ *PHARMACODYNAMICS ; Human ; Iodine Radioisotopes/DIAGNOSTIC USE ; Palate/ DRUG EFFECTS/EMBRYOLOGY/*PATHOLOGY ; Receptors, Endogenous Substances/ ANALYSIS ; Rubella/CONGENITAL/*PATHOLOGY SO - J Clin Invest 1986 May;77(5):1613-21 21 UI - 86195954 AU - Seyedin SM ; Thompson AY ; Bentz H ; Rosen DM ; McPherson JM ; Conti A ; Siegel NR ; Galluppi GR ; Piez KA TI - Cartilage-inducing factor-A. Apparent identity to transforming growth factor-beta. AB - Comparison of the sequence of the N-terminal 30 amino acids of cartilage-inducing factor-A (CIF-A) from bovine demineralized bone with the corresponding sequence of human transforming growth factor-beta (TGF-beta) revealed 100% identity. Furthermore, CIF-A stimulated normal rat kidney fibroblasts to become anchorage-independent and form colonies in soft agar (in the presence of epidermal growth factor) in a manner similar to TGF-beta. Similarly, TGF-beta from human platelets induced rat muscle mesenchymal cells to differentiate and synthesize cartilage-specific macromolecules in a manner equivalent to CIF-A. These data show that CIF-A and TGF-beta are closely related or identical molecules and that these factors may be involved in cell differentiation including cartilage formation as the first step in endochondral bone formation. MH - Amino Acid Sequence ; Animal ; Bone and Bones ; Cattle ; Cell Division/ DRUG EFFECTS ; Cell Line ; Chromatography, High Pressure Liquid ; Comparative Study ; *Growth Substances ; *Proteins/ISOLATION & PURIFICATION/PHARMACODYNAMICS SO - J Biol Chem 1986 May 5;261(13):5693-5 22 UI - 86177577 AU - Ibbotson KJ ; Harrod J ; Gowen M ; D'Souza S ; Smith DD ; Winkler ME ; Derynck R ; Mundy GR TI - Human recombinant transforming growth factor alpha stimulates bone resorption and inhibits formation in vitro. AB - Human recombinant transforming growth factor alpha (TGF alpha), which binds to the epidermal growth factor (EGF) receptor and causes several biological effects similar to those caused by EGF, was compared with murine EGF for its effects on a number of parameters of bone cell metabolism. TGF alpha stimulated bone resorption in two organ culture systems, the fetal rat long bone and neonatal mouse calvarial systems. TGF alpha stimulated bone resorption at concentrations as low as 0.1 ng/ml. TGF alpha effects on bone resorption in mouse calvariae were inhibited by indomethacin, suggesting that, like EGF, its effects were mediated by prostaglandin synthesis. TGF alpha had a different time course of action on bone resorption from that of EGF, causing more rapid release of previously incorporated 45Ca from bone cultures, suggesting that TGF alpha does not function on bone as a simple EGF analogue. TGF alpha also caused effects on osteoblast function resembling those of EGF. It inhibited alkaline phosphatase activity in cultured rat osteosarcoma cells with the osteoblast phenotype and inhibited collagen synthesis in fetal rat calvaria at concentrations of 1.0 ng/ml. The lowest concentration of TGF alpha (expressed as nanogram equivalents of EGF per ml) required to produce a response in all of the systems tested was about 1/10th of that needed for EGF to produce a similar effect. These results indicate that TGF alpha is a potent stimulator of bone resorption and inhibitor of bone formation as assessed by inhibition of collagen synthesis and alkaline phosphatase activity and are consistent with the hypothesis that TGF alpha may be responsible, at least in part, for the bone resorption associated with some tumors. MH - Alkaline Phosphatase/METABOLISM ; Animal ; Bone Resorption/*DRUG EFFECTS ; Collagen/BIOSYNTHESIS ; Comparative Study ; Dose-Response Relationship, Drug ; Epidermal Growth Factor-Urogastrone/*PHARMACODYNAMICS ; Indomethacin/PHARMACODYNAMICS ; Peptides/*PHARMACODYNAMICS ; Rats ; Recombinant Proteins/*PHARMACODYNAMICS ; Sarcoma, Osteogenic/ENZYMOLOGY ; Support, U.S. Gov't, P.H.S. ; Time Factors SO - Proc Natl Acad Sci USA 1986 Apr;83(7):2228-32 23 UI - 86161011 AU - Moseley JM ; Suva LJ TI - Molecular characterization of the EGF receptor and involvement of glycosyl moieties in the binding of EGF to its receptor on a clonal osteosarcoma cell line, UMR 106-06. AB - The epidermal growth factor receptor in cells of the UMR 106-06 clonal osteoblast line has been shown to be structurally similar to that previously characterized in other cell lines. A specific receptor component of approximately 165,000-185,000 Mr has been identified by polyacrylamide gel electrophoresis using the chemical crosslinker disuccinimidyl suberate to crosslink 125I-EGF to its receptor. Tunicamycin treatment of cells resulted in a dose-dependent loss of binding suggesting involvement of glycosyl moieties in EGF binding to its receptor. Competitive binding studies carried out using wheat germ lectin (WGL), concanavalin A (CON.A.), soybean lectin (SBL), and lentil lectin (ILL) to compete for binding of 125I-EGF revealed that CON A, WGL, and to a lesser extent LL could inhibit EGF binding; SBL was without effect. Treatment of the cells with neuraminidase which cleaves terminal sialic acid residues resulted in total loss of binding while alpha-glucosidase, beta-N-acetylglucosaminidase and alpha-mannosidase were without effect. These data indicate a specific interaction of EGF with terminal sialic acid residues of the EGF receptor. However, it would seem that the mannose residues which appeared to modify EGF binding were not available for the action of the above enzymes due to the presence of sialic acid. MH - Animal ; Binding Sites ; Binding, Competitive ; Bone and Bones/METABOLISM ; Clone Cells/METABOLISM ; Epidermal Growth Factor-Urogastrone/ *METABOLISM ; Kinetics ; Lectins/METABOLISM ; Molecular Weight ; Rats ; Receptors, Endogenous Substances/ISOLATION & PURIFICATION/*METABOLISM ; Sarcoma, Osteogenic/*METABOLISM ; Sialic Acids/METABOLISM ; Support, Non-U.S. Gov't SO - Calcif Tissue Int 1986 Feb;38(2):109-14 24 UI - 86202917 AU - Duursma SA ; Bijlsma JW ; van Paassen HC ; Slootweg MC TI - Oestrogens and bone metabolism: a hypothesis. AB - The explanations so far proposed for the effects of oestrogens on bone metabolism in post-menopausal women have been based on the changes seen in calciotrophic hormones. However, we demonstrated in a previous study [6] that there is a decrease in the concentration of somatomedin (an insulin-like growth factor) and an increase in that of growth hormone (GH) in oestrogen-treated post-menopausal women. On the basis of those findings and the relevant data published in the literature, we advance a hypothesis to the effect that the bone-metabolism changes that occur in post-menopausal women following oestrogen replacement therapy come about via changes in serum somatomedin and growth hormone concentrations. MH - Alkaline Phosphatase/BLOOD ; Bone and Bones/*METABOLISM ; Calcium/BLOOD/ URINE ; Ethinyl Estradiol/*METABOLISM ; Female ; Human ; Hydroxyproline/ URINE ; Menopause/DRUG EFFECTS ; Middle Age ; Phosphates/BLOOD ; Somatomedins/BLOOD ; Somatotropin/BLOOD SO - Maturitas 1986 Mar;8(1):1-6 25 UI - 86192132 AU - Canalis E ; Centrella M TI - Isolation of a nontransforming bone-derived growth factor from medium conditioned by fetal rat calvariae. AB - Previous studies have indicated that medium conditioned by 21-day-old fetal rat calvariae contains bioactive proteins termed bone-derived growth factors (BDGF) I and II. In the present studies we have purified the nontransforming BDGF II by dialysis, molecular sieving, three reverse phase HPLC steps, and preparative polyacrylamide gel electrophoresis. The second HPLC step (HPLC-2) yielded a recovery of 22% of the biological activity and achieved a 1500-fold purification, resulting in 20 micrograms protein/liter calvarial conditioned medium; the third HPLC step was of limited value in the purification of BDGF. Analytical PAGE revealed that the majority of the protein in HPLC-2-purified BDGF migrated with a relative molecular mass (Mr) of 11,000 and two additional proteins were seen at a Mr of 22,000-23,000. On preparative PAGE, the material migrating with a Mr of 11,000 stimulated parameters of bone and fibroblast growth in vitro, whereas the material with a Mr of 22,000-23,000 had less biological activity. Isoelectric focusing revealed that BDGF had an isoelectric point (pI) of 5. BDGF enhanced the incorporation of [3H]thymidine into DNA in fibroblast and calvarial cultures and of [3H]proline into collagen and noncollagen protein in calvariae. In conclusion, fetal rat calvariae secrete a BDGF with an estimated Mr of 11,000 and a pI of 5; this material stimulates bone and fibroblast growth in vitro. MH - Animal ; Biological Assay ; Bone and Bones/EMBRYOLOGY/*METABOLISM ; Cell Division ; Chromatography, High Pressure Liquid ; Collagen/BIOSYNTHESIS ; Culture Media/ANALYSIS ; DNA/BIOSYNTHESIS ; Electrophoresis, Polyacrylamide Gel ; Fibroblasts/METABOLISM ; Growth Substances/ *ISOLATION & PURIFICATION/PHARMACODYNAMICS ; Isoelectric Focusing ; Molecular Weight ; Rats ; Support, U.S. Gov't, P.H.S. ; Tissue Culture SO - Endocrinology 1986 May;118(5):2002-8 26 UI - 86184127 AU - Isgaard J ; Nilsson A ; Lindahl A ; Jansson JO ; Isaksson OG TI - Effects of local administration of GH and IGF-1 on longitudinal bone growth in rats. AB - The effect of local administration of growth hormone (GH) and insulinlike growth factor 1 (IGF-1) on longitudinal bone growth was studied in the proximal tibia of hypophysectomized rats, by using the tetracycline method. Human GH (hGH) stimulated local bone growth when administered into the epiphysial growth plate, into the epiphysis through an implanted cannula, or into the knee joint intraarticularly. In contrast, hGH administration into the metaphysis did not cause such a stimulation. The effect of hGH was dose dependent, and the lowest daily dose of hGH that caused a stimulation was 50 ng. hGH produced by cloned bacteria was as effective as pituitary-derived hGH, excluding the possibility of a pituitary growth factor being the active compound. GH from other mammalian species (rat GH, ovine GH, and bacterially produced bovine GH) also stimulated local bone growth. Ovine prolactin (oPRL) stimulated local bone growth but the threshold dose of oPRL was approximately 100 times higher than that of hGH, suggesting that contamination of this preparation by GH may account for the stimulation. Reduced carboxymethylated human GH, that has a greatly reduced anabolic activity, did not stimulate local bone growth. Local administration of 5 micrograms of bacterially produced human IGF-1 per day produced a small but significant effect on unilateral bone growth. Simultaneous administration of hGH had no additive effect with, nor did it potentiate, the stimulatory effect of IGF-1. The present study confirms and extends earlier investigations, showing that local injection of GH at the site of the epiphysial growth plate stimulates unilateral bone growth. The study also shows that local administration of IGF-1 stimulates longitudinal bone growth. MH - Animal ; Bone Development/*DRUG EFFECTS ; Cattle ; Dose-Response Relationship, Drug ; Epiphyses ; Human ; Hypophysectomy ; Insulin-Like Growth Factor I/ADMINISTRATION & DOSAGE/*PHARMACODYNAMICS ; Knee Joint ; Male ; Rats ; Rats, Inbred Strains ; Sheep ; Somatomedins/ *PHARMACODYNAMICS ; Somatotropin/ADMINISTRATION & DOSAGE/ *PHARMACODYNAMICS ; Support, Non-U.S. Gov't SO - Am J Physiol 1986 Apr;250(4 Pt 1):E367-72 27 UI - 86183910 AU - Kaufmann S ; Jones M ; Culler FL ; Jones KL TI - Growth hormone deficiency in the Rothmund-Thomson syndrome. AB - We describe the first proven occurrence of growth hormone deficiency in an individual with the Rothmund-Thomson syndrome. This was suspected because of the patient's severely retarded growth and bone age and her failure to respond normally to growth hormone stimulation testing with l-DOPA, arginine, and growth hormone releasing factor. In addition, we have briefly reviewed other genetic and malformation syndromes that have been found associated with growth hormone deficiency. We recommend that growth hormone deficiency be considered in these syndromes, especially when the growth failure is more marked than expected. MH - Arginine/DIAGNOSTIC USE ; Case Report ; Child ; Dwarfism/ETIOLOGY/ FAMILIAL & GENETIC ; Female ; Human ; Insulin-Like Growth Factor I/BLOOD ; Levodopa/DIAGNOSTIC USE ; Poikiloderma Congenitale/FAMILIAL & GENETIC/ *METABOLISM ; Skin Diseases/*METABOLISM ; Somatotropin/*DEFICIENCY ; Somatotropin Releasing Hormone/DIAGNOSTIC USE ; Support, U.S. Gov't, P.H.S. SO - Am J Med Genet 1986 Apr;23(4):861-8 28 UI - 86164152 AU - Boepple PA ; Mansfield MJ ; Wierman ME ; Rudlin CR ; Bode HH ; Crigler JF Jr ; Crawford JD ; Crowley WF Jr TI - Use of a potent, long acting agonist of gonadotropin-releasing hormone in the treatment of precocious puberty. AB - Studies utilizing the administration of GnRH in various GnRH-deficient models have revealed the critical importance of the dose and mode of delivery of this releasing factor in determining the subsequent pituitary response. Chronic administration of long acting GnRH agonists (GnRHa), like continuous infusion of high doses of the native peptide, results in suppression of pituitary gonadotropin secretion. This selective and reversible suppression of gonadotropin secretion suggested several therapeutic applications for these analogs, particularly in the treatment of central precocious puberty (CPP), a disorder for which the previously available therapies lacked uniform efficacy and were associated with potential side effects. In our series, 74 children with CPP have been treated during the last 5 yr with the potent GnRH agonist, [D-Trp6, Pro9-ethylamide(NEt)]GnRH. Having selected a dose and route of administration that produced uniform suppression of spontaneous and stimulated pituitary gonadotropin secretion, GnRHa therapy resulted in a fall of gonadal sex steroid levels into the prepubertal range, a halting or regression of secondary sexual development, and a complete cessation of menses. Growth velocity slowed during therapy, with this slowing more pronounced during prolonged treatment periods and among those patients with more advanced chronological and skeletal ages. Skeletal maturation was retarded to a greater degree than linear growth, with resultant increases in the predictions for adult stature. Moreover, these benefits have been achieved in the absence of significant side effects. Complete reversal of the suppression of gonadarche has followed discontinuation of therapy; however, patterns of growth and skeletal maturation after discontinuation of GnRHa administration remain to be characterized. Thus, the impact of GnRHa therapy on final height must await further longitudinal study. The selective nature of GnRHa suppression of gonadarche also permits an investigation of the natural history of adrenarche and its discrete influences upon skeletal growth and maturation. In addition, GnRHa therapy of CPP provides a unique opportunity to study the effects of gonadal sex steroids on GH secretion and somatomedin-C (Sm-C) generation during sexual maturation. Finally, the detailed characterization of children with precocious puberty has helped to define more precisely a subset of patients whose precocity occurs in the absence of demonstrable gonadotropin secretion.(ABSTRACT TRUNCATED AT 400 WORDS) MH - Adrenal Glands/PHYSIOLOGY ; Adrenal Hyperplasia, Congenital/COMPLICATIONS ; Bone Development ; Child ; Child, Preschool ; Dehydroepiandrosterone/ ANALOGS & DERIVATIVES/BLOOD ; Estradiol/BLOOD ; Female ; Hamartoma/ COMPLICATIONS ; Human ; Hypothalamic Neoplasms/COMPLICATIONS ; Insulin-Like Growth Factor I/SECRETION ; LH/SECRETION ; LH-FSH Releasing Hormone/*ANALOGS & DERIVATIVES/ADMINISTRATION & DOSAGE/ADVERSE EFFECTS/ THERAPEUTIC USE ; Male ; Pituitary Hormone Releasing Hormones/PHYSIOLOGY/ SECRETION ; Puberty, Precocious/*DRUG THERAPY/ETIOLOGY/PHYSIOPATHOLOGY ; Review ; Somatotropin/SECRETION ; Support, U.S. Gov't, P.H.S. ; Testosterone/BLOOD SO - Endocr Rev 1986 Feb;7(1):24-33 29 UI - 86135772 AU - Boland CJ ; Fried RM ; Tashjian AH Jr TI - Measurement of cytosolic free Ca2+ concentrations in human and rat osteosarcoma cells: actions of bone resorption-stimulating hormones. AB - Influx of extracellular Ca++ into bone cells has been postulated as an early action of PTH and other bone resorption-stimulating factors. To test this hypothesis directly, we measured the cytosolic free Ca2+ concentration ([Ca2+]i) in two hormone-responsive human (SaOS-2 and G-292) and two rat osteosarcoma cell lines (Ros 25/1 and Ros 17/2.8) and in primary cultures of bone cells from neonatal mouse calvaria using the fluorescent Ca2+ indicator Quin 2. Actions of bovine PTH-(1-34), vasoactive intestinal peptide, epidermal growth factor, prostaglandin E2, and ionomycin were studied. Medium cAMP (20 min; 37 C; 25 microM 3-isobutyl-1-methylxanthine) was quantitated by RIA. Basal [Ca2+]i was: SaOS-2, 126 +/- 8 nM; G-292, 61 +/- 6 nM; Ros 25/1, 109 +/- 15 nM; Ros 17/2.8, 363 +/- 42 nM; and primary cultures, 266 +/- 39 nM (mean +/- SE; n = 3-14). In each cell type, no acute (1 sec to 20 min) spike in [Ca2+]i was observed in response to PTH (24-120 nM), vasoactive intestinal peptide (100 nM), epidermal growth factor (17 nM), or prostaglandin E2 (2.8 microM). However, in SaOS-2 cells only, PTH reproducibly increased [Ca2+]i 10-15% above basal values beginning about 3 min after hormone addition, and this small increase returned to baseline at 15-20 min. Ionomycin (100 nM) elicited an immediate spike in [Ca2+]i to levels 2- to 4-fold above basal in all cells; the peak [Ca2+]i decayed rapidly (within 4-5 min) to baseline in G-292, Ros 25/1, and Ros 17/2.8 cells. The decay of peak [Ca2+]i in SaOS-2 was prolonged. To test for intact hormone responses in Quin 2-loaded cells, cAMP accumulation was measured. In SaOS-2 and Ros 17/2.8, both control and Quin 2-loaded cells showed similar increases in cAMP in response to PTH. Considering the limitations of the Quin 2 technique, we conclude that in the four hormone-responsive bone cell lines and primary cultures of bone cells tested, acute elevation of [Ca2+]i is not an inevitable consequence of receptor occupancy and/or adenylate cyclase activation by bone resorption-stimulating hormones. MH - Adenosine Cyclic Monophosphate/METABOLISM ; Aminoquinolines/DIAGNOSTIC USE ; Animal ; Bone Resorption/DRUG EFFECTS ; Calcium/*METABOLISM ; Cell Line ; Cells, Cultured ; Cytosol/METABOLISM ; Epidermal Growth Factor-Urogastrone/PHARMACODYNAMICS ; Ethers/PHARMACODYNAMICS ; Human ; Membrane Potentials ; Mice ; Parathyroid Hormones/PHARMACODYNAMICS ; Prostaglandins E/PHARMACODYNAMICS ; Rats ; Sarcoma, Osteogenic/ *METABOLISM ; Support, U.S. Gov't, P.H.S. ; Vasoactive Intestinal Peptide/ PHARMACODYNAMICS SO - Endocrinology 1986 Mar;118(3):980-9 30 UI - 86125923 AU - Thorngren KG ; Hallengren B TI - Bioassayable growth hormone activity in blood from healthy individuals and acromegalic patients. AB - Biological growth activity (bioassayable GH) was determined in blood from healthy individuals and from patients with acromegaly using the rate of longitudinal bone growth in hypophysectomized rats with tetracycline as intravital marker. Also radioimmunoassayable GH and somatomedin A activity were determined. In pooled plasma or serum from normal subjects no bioassayable growth activity was demonstrated. In the clinically active acromegalic patients as a group as well as in one individual patient there was a significant (P less than 0.05) bioassayable growth activity in serum as compared to serum from normal subjects. The bioassay determination of GH in plasma/serum from normal subjects and acromegalic patients was hampered by the toxicity and the problems connected with the administration of large volumes. MH - Acromegaly/*BLOOD ; Adult ; Aged ; Animal ; Body Weight ; Bone Development ; Female ; Heart/ANATOMY & HISTOLOGY ; Human ; Hypophysectomy ; Male ; Middle Age ; Organ Weight ; Radioimmunoassay ; Radioligand Assay ; Rats ; Somatomedins/BLOOD ; Somatotropin/*BLOOD ; Support, Non-U.S. Gov't SO - Acta Endocrinol (Copenh) 1986 Jan;111(1):3-9 31 UI - 86118707 AU - Bertolini DR ; Nedwin GE ; Bringman TS ; Smith DD ; Mundy GR TI - Stimulation of bone resorption and inhibition of bone formation in vitro by human tumour necrosis factors. AB - When leukocytes are exposed to mitogens or antigens in vitro, they release bone-resorbing activity into the culture supernatants which can be detected by bioassay. Like many lymphocyte-monocyte products, this activity has been difficult to purify because of its low abundance in activated leukocyte cultures and the unwieldy bioassay required to detect biological activity. Partially purified preparations of this activity inhibit bone collagen synthesis in organ cultures of fetal rat calvariae. Recent data suggest that both activated lymphocytes and monocytes release factors which could contribute to this activity. Recently, monocyte-derived tumour necrosis factor alpha (TNF-alpha) and lymphocyte-derived tumour necrosis factor beta (TNF-beta) (previously called lymphotoxin), two multifunctional cytokines which have similar cytotoxic effects on neoplastic cell lines, have been purified to homogeneity and their complementary DNAs cloned and expressed in Escherichia coli. As both of these cytokines are likely to be present in activated leukocyte supernatants, we tested purified recombinant preparations for their effects on bone resorption and bone collagen synthesis in vitro, and report here that both cytokines at 10(-7) to 10(-9) M caused osteoclastic bone resorption and inhibited bone collagen synthesis. These data suggest that at least part of the bone-resorbing activity present in activated leukocyte culture supernatants may be due to these cytokines. MH - Animal ; Bone Development/*DRUG EFFECTS ; Bone Resorption/*DRUG EFFECTS ; Cells, Cultured ; Escherichia Coli/GENETICS ; Fetus ; Glycoproteins/ GENETICS/*PHARMACODYNAMICS ; Growth Inhibitors/*PHARMACODYNAMICS ; Human ; Rats ; Recombinant Proteins/*PHARMACODYNAMICS ; Support, U.S. Gov't, P.H.S. SO - Nature 1986 Feb 6-12;319(6053):516-8 32 UI - 86109201 AU - Heinze E ; Brenner R ; Nguyen-Thi C ; Vetter U ; Leupold D ; Pohlandt F TI - Skeletal growth in fetal rats. Effects of glucose and amino acids. AB - The modified hyperglycemia-hyperinsulinism hypothesis, which characterizes intrauterine growth of diabetic pregnancy, was studied in fetal rats. From day 19 to day 21 postconception, pregnant rats were constantly infused with saline, amino acids, or glucose. In the fetus, serum somatomedin activity was determined, with the porcine bioassay and the incorporation of 3H-thymidine into rib cartilage and isolated chondrocytes in vivo in response to serum from normal maternal or fetal rats. In comparison with control fetuses, body weights were decreased in glucose-exposed fetuses (4.66 +/- 0.25 versus 3.75 +/- 0.99, N = 121; P less than 0.001), and increased (4.87 +/- 0.57, N = 105; P less than 0.05) in amino acid-exposed fetuses. Serum somatomedin activity (U/ml) was higher in glucose-treated (0.79 +/- 0.40, N = 11; P less than 0.05) and amino acid-treated animals (0.90 +/- 0.16, N = 10; P less than 0.001) than in controls (0.55 +/- 0.04, N = 13). In vivo labeling with thymidine resulted in a higher radioactivity of cartilage in small fetuses compared with large fetuses when the dams had been infused with saline (r = -0.531, N = 56; P less than 0.001) or amino acids (r = -0.292, N = 52; P less than 0.01). Opposite results were obtained in hyperglycemic animals (r = 0.542, N = 54; P less than 0.001). When isolated chondrocytes were incubated with serum from normal fetal rats, the incorporation of thymidine was about 10 times higher into cells from small fetuses than from large fetuses, irrespective of the infusion regimen.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Amino Acids/BLOOD/*PHARMACODYNAMICS ; Animal ; Blood Glucose/ANALYSIS ; Body Weight/DRUG EFFECTS ; Bone and Bones/DRUG EFFECTS/*EMBRYOLOGY ; Female ; Fetus/DRUG EFFECTS/PHYSIOLOGY ; Glucose/*PHARMACODYNAMICS ; Insulin/PHARMACODYNAMICS ; Male ; Pregnancy ; Rats ; Rats, Inbred Strains ; Somatomedins/BLOOD ; Support, Non-U.S. Gov't SO - Diabetes 1986 Feb;35(2):222-7