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Biomedical subjects

M T Story

Publications and source records attributed to M T Story.

At least 19 recordsLinked to original sources

Relationships between vitamin and mineral supplement use, dietary intake, and dietary adequacy among adolescents.

OBJECTIVE: To examine patterns of supplement use among US adolescents and the relationship between supplement use and dietary intake and adequacy. DESIGN: Adolescents self-reported 2 days of food intake using the 24-hour recall method and supplement use during a personal interview conducted as part of the 1994 Continuing Survey of Food Intakes of Individuals (CSFII). SUBJECTS: A national sample of 423 adolescents included in the 1994 CSFII survey. STATISTICAL ANALYSIS: chi 2 analysis was used to determine which demographic factors were significantly related to patterns of supplement use. Weighted percentages of adolescents by category of supplement use for selected vitamins and minerals (calcium; iron; zinc; folic acid; and vitamins A, B-6, C, and E) are presented. Relationships between dietary intake of macronutrients and vitamins and minerals among adolescents and supplement use were determined using a least-squares model of general linear regression. RESULTS: Approximately one-third of adolescents reported using supplements, with 15.6% of youth using them on a daily basis. The majority of supplement users reported taking multivitamins (N = 95; 65.5%) whereas only one-third of supplement users reported taking individual vitamins or minerals. Supplement use was found to vary by gender, household size, and US region of residence. Adolescents who reported using supplements had higher mean dietary intakes of most micronutrients and lower intakes of total and saturated fat than those who did not use supplements. More than one-third of adolescents had dietary intakes of vitamins A and E, calcium, and zinc that were < 75% of the US Recommended Dietary Allowance. APPLICATIONS/CONCLUSIONS: The majority of US adolescents do not use vitamin or mineral supplements. Interestingly, adolescents who do use supplements, even on an infrequent basis, consume diets that are more nutrient-dense than those who do not use supplements. Dietary intakes of several micronutrients were inadequate among all adolescents in this study, regardless of supplement use status. There is a need to develop and implement programs aimed at improving the dietary intakes of US adolescents.

Adolescent↗

Characteristics of antigens from an HCG alpha secreting cell line (ElCo) derived from human breast carcinoma in a Native American patient.

OBJECTIVE: Our purpose is to report a cell line derived from a Native American patients that could serve as a model for ethnic diversity in cell culture research. STUDY DESIGN: Tumor biopsy specimens from reproductive tract malignancies were explanted in the cell culture laboratory. Characterization of a successfully established line included a parameter commonly observed in minority populations in rapid moving Type A glucose-6-phosphate dehydrogenase enzyme (G6-PD). Multiple other standard characterization studies were carried out. RESULTS: A breast cancer cell line with a G6-PD-A enzyme was derived from an American Indian patient; this pattern is commonly observed in African-American populations. Results of lymphocyte-mediated cytotoxicity tests indicated that lymphocytes from patients with breast cancer were cytotoxic to ElCo cells. On the other hand, lymphocytes from patients with other types of cancer were not significantly different from healthy donor lymphocytes in their cytotoxicity. These studies indicate that ElCo cells express tumor-associated antigen(s). Many antigens of tumor cell origin were detected with use of antisera raised in rabbits in concentrates of ElCo cell culture fluid. One of these antigens was the alpha subunit of human chorionic gonadotropin. CONCLUSIONS: Chemotherapy testing, vaccine production, and various new treatment schemes are most commonly developed in cell culture systems. These systems should reflect characteristics of the target population so that desired outcomes best fit the population being served. Thus the cell line being reported, which was derived from a Native American woman, represents a tool that can be used in cell culture research as a model for diversity.

Animals↗

Regulation of basic fibroblast growth factor expression by transforming growth factor beta in cultured human prostate stromal cells.

Basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGFbeta1) are potential autocrine growth regulators of the prostatic stroma, and therefore may play a role in the development of benign prostatic hyperplasia (BPH). We reported that TGFbeta1 increased bFGF and bFGF mRNA expression in cultured human prostate stromal cells (PS). The current study extends those studies and investigates the mechanism by which TGFbeta1 upregulates the level of bFGF mRNA. A solution hybridization assay was used to quantitate bFGF mRNA. Treatment of PS for 6 hr with TGFbeta1 (1 ng/ml) maximally stimulated bFGF mRNA expression. TGFbeta2 and TGFbeta3 were similarly active in upregulating bFGF mRNA. TGFbeta1 or cycloheximide each increased bFGF mRNA about 3-fold. The effect of these agents was not additive. This suggested that a labile protein was involved in processing bFGF mRNA. Determination of message stability indicated that the half-life of bFGF mRNA in TGFbeta1-treated PS was 6.8 hr, as compared to 4.3 hr in untreated cells. The data indicated that posttranscriptional mechanisms that increased message stability were, at least in part, responsible for upregulation of bFGF mRNA by TGFbeta1 in PS. Our studies suggest that growth of the prostatic stroma is regulated by the interaction of members of two families of growth modulators, bFGFand TGFbeta. It remains to be determined if an imbalance in this system in favor of stroma hyperplasia plays a role in the development of BPH.

Blotting, Northern↗

Expression of transforming growth factor beta 1 (TGF beta 1), -beta 2, and- beta 3 by cultured human prostate cells.

Transforming growth factor beta s (TGF beta s) are members of a superfamily of polypeptides that control cell cycle progression and a variety of other cellular activities. TGF beta family members, -beta 1, -beta 2, and -beta 3, have been identified in prostate. The levels of expression of these TGF beta isotypes have been reported to vary with the pathologic state of the prostate. While the significance of these observations remains to be elucidated there is little doubt that TGF beta s play an important role in controlling growth of the prostate. The prostatic cells expressing TGF beta s have not been identified. This information would provide insight into the physiologic role of TGF beta s and suggest ways that growth control may be altered in prostate disease. We used stromal (PS) and epithelial (PE) cells, cultured from normal human prostate and benign prostatic hyperplasia (BPH), to study the effect of TGF beta s on cell proliferation and TGF beta transcript and protein expression. The proliferation of PS and PE was inhibited by pM quantities of TGF beta 1, -beta 2, and -beta 3. Both cell types expressed transcripts for all three TGF beta isotypes, but PS primarily secreted TGF beta 1, whereas PE secreted more TGF beta 2 than TGF beta 1. These observations suggest that TGF beta s are antiproliferative agents in vivo, and that the stroma is the source of TGF beta 1 while the epithelium is the major source of TGF beta 2 in prostate. There were no significant differences in the growth response to TGF beta s, the TGF beta-isotype expressed, or the amount of TGF beta secreted by cells cultured from normal prostate or BPH.

Cell Division↗

Regulation of prostate growth by fibroblast growth factors.

Growth of the prostate is controlled by androgen. However, there is information indicating that androgen may not act directly, but may act indirectly through polypeptide growth factors, to control prostate growth. This review will focus on the involvement of members of the fibroblast growth factor (FGF) family in this process. The properties of FGFs and FGF-receptors are described that implicate these molecules in growth control. Information is provided that prostate stromal cells synthesize FGF2 and FGF7. FGF2 is a potent mitogen for stromal cells; whereas, FGF7 is exclusively a mitogen for epithelial cells. Transforming growth factor beta (TGF beta), also produced by prostate cells, inhibit cell growth. This suggests that prostate growth is controlled by autocrine and paracrine mechanisms. Evidence is presented that altered FGF expression accompanies benign prostatic hyperplasia and prostate cancer. A model is proposed whereby androgen regulates TGF beta, influencing FGF2 and FGF7 expression, and in turn regulating growth of the prostatic stroma and epithelium. An imbalance in the influence of these growth factors may contribute to prostate disease.

Animals↗

Regional concentration of basic fibroblast growth factor in normal and benign hyperplastic human prostates.

Basic fibroblast grown factor (bFGF) is a potent mitogen for mesenchymal cells, including fibroblasts cultured from prostate, and has been postulated to play a role in the development of benign prostatic hyperplasia (BPH). If this is the case, it might be expected that bFGF levels would be elevated in the adenomas of BPH and in the periurethral region of the prostate where BPH is believed to arise. This study was undertaken to test this hypothesis. The concentration of bFGF was evaluated in 31 prostates, 13 normal glands and 18 with BPH. A method for quantitating bFGF by radioimmunoassay was developed that enabled growth factor levels to be correlated to the geographic region of the prostate and the histopathology of the specimen. A 2- to 3-fold higher concentration of bFGF (ng./g. of tissue) was noted in the benign hyperplastic prostates when compared with the adult normal glands. Pubertal specimens demonstrated low growth factor levels comparable to those observed in the normal adult group. Two prepubertal prostates analyzed had high levels similar to those measured in the hyperplastic glands. While the levels of bFGF in the normal adult prostates were highest in the periurethral region, statistical analysis failed to demonstrate a significant difference. Similarly, quantitative morphometric evaluation failed to demonstrate any significant differences in bFGF concentration related to the proportion of stromal, epithelial, or lumenal elements in the tissue sections.

Adolescent↗

Characteristics of FGF-receptors expressed by stromal and epithelial cells cultured from normal and hyperplastic prostates.

Three fibroblast growth factors (FGFs), acidic FGF (FGF1), basic FGF (FGF2), and keratinocyte growth factor (FGF7) have been identified in prostate. To understand how FGFs regulate growth of the prostate, and to determine if regulation is altered in benign prostatic hyperplasia (BPH), the mitogenic potential of FGFs, receptor binding, and FGF-receptor (FGFR) gene expression of stromal (PS) and epithelial cells (PE) cultured from normal human prostate and BPH where determined. FGF1 and FGF2, but not FGF7, were mitogens for PS. FGF1 and FGF7 were potent mitogens for PE, but FGF2 was a weak mitogen for these cells. Both PS and PE exhibited high affinity binding (pM K) of iodinated-FGF2. The K was 4-fold and 12-fold higher for PS than for PE cultured from normal prostate and BPH, respectively. Northern analysis indicated that PS, but not PE, expressed FGFR type 1 (FGFR1) mRNA. The reverse transcriptase polymerase chain reaction (RT-PCR) was used to evaluate FGFR type 2 (FGFR2) expression. The size of amplified DNA fragments, and nucleotide sequences, indicated that PS also expressed transcripts for the exon IIIc RNA splice variant of FGFR2. A RT-PCR product with the FGFR2 exon IIIb nucleotide sequence joined with the exon IIIc sequence was amplified with poly A+ RNA from PE and primers spanning both exons. Thus, PE did not alternatively splice mRNA for FGFR2 exon IIIb and exon IIIc. No differences in the mitogenic potential of FGFs, receptor binding (K or number of sites), or FGFR gene expression were found in cells cultured from normal prostate and BPH.

Base Sequence↗

Influence of transforming growth factor beta 1 and other growth factors on basic fibroblast growth factor level and proliferation of cultured human prostate-derived fibroblasts.

Basic fibroblast growth factor (bFGF) has been identified in the human prostate. The level of bFGF has been reported to be elevated in benign prostatic hyperplasia (BPH), compared with normal prostate, suggesting that the growth factor may play a role in this disease of the prostate. Basic FGF is a mitogen for cultured human prostate-derived fibroblasts (PF). PF also synthesize bFGF, suggesting that growth regulation of these cells may be under autocrine control. The current study was undertaken to identify factors that affect PF proliferation and bFGF expression. Transforming growth factor beta 1 (TGF-beta 1) inhibited PF proliferation. The inhibition by TGF-beta 1 was partially overcome by bFGF but not by epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor type 1 (IGF-1), or insulin. Incubation of PF with TGF-beta 1 increased bFGF mRNA and immunoreactive bFGF levels in a dose- and time-dependent fashion. None of the other growth factor studies affected bFGF levels. PF were also found to express TGF-beta 1 mRNA, the level of which was increased two- to fivefold by TGF-beta 1. These observations suggest that PF proliferation is controlled by the interaction of two different growth factors. It is possible that bFGF/TGF-beta imbalance in favor of cell proliferation promotes prostatic stromal hyperplasia.

Blood Proteins↗

Polypeptide modulators of prostatic growth and development.

Normal and abnormal developmental events in the prostate are strongly influenced by androgens. There is abundant evidence, however, that androgens are not the only substances present that have the capacity to influence prostatic growth. A number of polypeptides that either stimulate or inhibit growth have now been identified in the prostate. These include members of the HBGF family, TGF-beta family, EGF and TGF-alpha, PDGF, NGF, and the less well characterized osteoblast growth factors. In some cases, the prostatic cell population, stromal or epithelial, that synthesizes the growth factor and its receptor is known. This information and the properties of the growth factors suggest ways in which these polypeptides may be involved in regulating growth of the prostate, including benign prostatic hyperplasia and prostate cancer.

Bone Neoplasms↗

Cultured human prostate-derived fibroblasts produce a factor that stimulates their growth with properties indistinguishable from basic fibroblast growth factor.

Fibrostromal proliferation is believed to be important in the development of benign prostatic hyperplasia (BPH). We found that a mitogen for cultured mesodermal-derived cells was present in extracts of BPH tissue. The mitogen was identified as basic fibroblast growth factor (bFGF). Previous studies did not determine the cell population(s) responsible for bFGF production in the prostate. This information is important to the understanding of the role of bFGF in the etiology of BPH. Human prostate-derived fibroblasts (PF) were initiated in culture. Recombinant bFGF and PF lysates stimulated tritiated thymidine uptake by quiescent PF cells. Greater than 90% of the mitogen in PF lysates bound to heparin-Sepharose and had the same elution profile and apparent molecular weight as bFGF isolated from BPH tissue. The growth factor in PF lysates competed with recombinant iodinated bFGF for binding to antiserum to (1-24)bFGF. Cultured PF incorporated 35S-methionine into protein that was precipitated by antiserum to bFGF. The apparent molecular weight of the radiolabeled protein, about 17,000, was similar to authentic bFGF. The observations are consistent with the interpretation that cultured PF synthesize a growth factor that stimulates their growth with properties that are indistinguishable from bFGF.

Cell Survival↗

Autonomic control of acid phosphatase exocrine secretion by the rat prostate.

In vivo prostatic secretion was collected from retired breeder Sprague Dawley rats using a method for isolated perfusion of the rat prostatic urethra. Enzymatic acid phosphatase determination was performed on the collected effluent. Control acid phosphatase secretion was 24.2 +/- 2.7 nm over 30 minutes. Intravenous phenylephrine 5 mg/kg stimulated a 10 fold increase in acid phosphatase secretion. The secretion seen with phenylephrine was dose dependent and could be blocked with prazosin, but not yohimbine, atropine, or propranolol. Intravenous beta-adrenergic agonist isoproterenol caused no increase in the secretion of rat prostatic acid phosphatase. Intravenous administration of the cholinergic agonist pilocarpine also resulted in a dose dependent rise in acid phosphatase secretion. The stimulation seen could be blocked by atropine but not phentolamine or propranolol. The stimulation of acid phosphatase secretion seen with alpha 1 adrenergic or cholinergic agonists was not additive. Intravenous vasoactive intestinal peptide did not stimulate acid phosphatase secretion nor did it augment the secretion induced by alpha 1 adrenergic or cholinergic agonists. Release of acid phosphatase into rat prostatic exocrine secretion is under both alpha 1 adrenergic and cholinergic control.

Acid Phosphatase↗

Cultured human foreskin fibroblasts produce a factor that stimulates their growth with properties similar to basic fibroblast growth factor.

To determine if fibroblasts could be a source of fibroblast growth factor (FGF) in tissue, cells were initiated in culture from newborn human foreskin. Cells were studied in Passages 2 to 8. Fibroblast cell lysates promoted radiolabeled thymidine uptake by cultured quiescent fibroblasts. Seventy-nine percent of the growth-promoting activity of lysates was recovered from heparin-Sepharose. The heparin-binding growth factor reacted on immunoblots with antiserum to human placenta-derived basic FGF and competed with iodinated basic FGF for binding to antiserum to (1-24)bFGF synthetic peptide. To confirm that fibroblasts were the source of the growth factor, cell lysates were prepared from cells incubated with radiolabeled methionine. Heparin affinity purified material was immunoprecipitated with basic FGF antiserum and electrophoresed. Radiolabeled material was detected on gel autoradiographs in the same molecular weight region as authentic iodinated basic FGF. The findings are consistant with the notion that cultured fibroblasts express basic FGF. As these cells also respond to the mitogen, it is possible that the regulation of their growth is under autocrine control. Fibroblasts may be an important source of the growth factor in tissue.

Aging↗

Exocrine secretion of epidermal growth factor by the rat prostate: effect of adrenergic agents, cholinergic agents, and vasoactive intestinal peptide.

Perfusion of the rat prostatic urethra in vivo provided a means of collecting the rat prostatic secretory product. The secretion of the protein epidermal growth factor (EGF) was investigated by radioimmunoassay (RIA). Baseline secretion of EGF into prostatic fluid was less than .03 +/- .004 SEM ng/min. The alpha-adrenergic agonist phenylephrine caused an increase in EGF to 3.6 +/- .4 SEM ng per 30-min period. The stimulation was blocked completely by prazosin and only partially by yohimbine, indicating primarily alpha 1 control. One mg/kg IV phenylephrine produced a maximal response. The beta-adrenergic agonist isoproterenol caused no increased secretion of EGF. The cholinergic agonist pilocarpine stimulated EGF secretion to 3.2 +/- .6 SEM ng per 30-min period. Atropine blocked the cholinergic stimulation. The combination of phenylephrine and pilocarpine did not result in greater stimulation than either agent alone. Vasoactive intestinal peptide (VIP) did not stimulate EGF secretion, nor did it augment either pilocarpine or phenylephrine-stimulated secretion. EGF secretion into rat prostatic fluid is under both alpha 1-adrenergic and cholinergic control.

Animals↗

A growth factor in bovine and human testes structurally related to basic fibroblast growth factor.

Homogenates of human testes, epididymides and prostate, and calf testes and epididymides are mitogenic for cultured human foreskin fibroblasts. The growth factors appear similar in that they are inactivated by boiling and acid, but not by treatment with reducing agent. The growth factor in human and bovine testes was partially purified from tissue homogenates, prepared in high ionic strength buffer (pH 7.6) containing protease inhibitors, by ammonium sulfate precipitation and two cycles of heparin-Sepharose chromatography. The growth factor in calf testes was also partially purified from tissue extracted in ammonium sulfate without protease inhibitors, acidified to pH 4.5, and precipitated by ammonium sulfate followed by two cycles of heparin-affinity chromatography. A predominant 17,500 molecular weight (MW) growth factor was identified from alkaline homogenates of human and calf testes by its reactivity with antisera prepared against synthetic peptides whose sequences corresponded to residues 1-12 (amino-terminal), 33-43 (internal) and 136-145 (carboxy-terminal) of bovine basic fibroblast growth factor (bFGF). A slightly smaller 16,600 MW peptide from acidic extracts of calf testes also reacted with antisera to the three synthetic peptides. A 15,500 MW peptide, lacking immunoreactivity with antiserum to the amino-terminal synthetic peptide, was also seen. These findings suggest that a growth factor is present in human and calf testes that is structurally related to bFGF. The structure of the growth factors appears to be altered during the isolation procedure.

Aged↗

Characterization of growth factors derived from the rat ventral prostate.

Tissue homogenates of rat ventral prostate were examined for growth factor activity using a fibroblast mitogenesis assay. G-75 Sephadex gel filtration separated the growth factor activity into two peaks. A broad first peak contained 98% of the protein and several growth factor moieties. A smaller second peak (MW 6,000) contained epidermal growth factor (EGF) as determined by binding in both a competitive receptor binding assay and a radioimmunoassay using anti-mouse epidermal growth factor (anti-mEGF). The broad first peak also contained substantial amounts of EGF-like activity as higher MW forms of EGF. The broad first peak was further fractionated by heparin-Sepharose affinity chromatography. A major fraction with growth factor activity eluted at 1.5 M NaCl and this fraction was shown to contain bFGF by immunostaining with antisera prepared against synthetic peptides corresponding to amino acid sequences 1-12 (amino terminal), 33-43 (internal), and 136-145 (carboxy terminal) of basic fibroblast growth factor (bFGF). EGF-like and bFGF-like molecules account for the major growth factor activity in the rat ventral prostate.

Animals↗

Prostatic growth factor: purification and structural relationship to basic fibroblast growth factor.

Prostatic growth factor (PrGF) was purified from alkaline homogenates of human benign prostatic hyperplastic tissue by a combination of ammonium sulfate precipitation, heparin affinity chromatography, and cation-exchange chromatography. The 17,600-dalton, basic (pI 10.2) PrGF is related to basic fibroblast growth factor (bFGF) since antisera raised against synthetic peptides with sequence homologies corresponding to an internal peptide and amino- and carboxyl-terminal peptides of bFGF react with the growth factor. The growth factor appears larger than bFGF, suggesting that additional amino-terminal sequences may be present as a result of alkaline extraction in the presence of protease inhibitors.

Amino Acid Sequence↗

Amino-terminal sequence of a large form of basic fibroblast growth factor isolated from human benign prostatic hyperplastic tissue.

Homogenization of human benign prostatic hyperplastic tissue in high ionic strength alkaline buffer containing protease inhibitors resulted in the isolation of a 17,400 molecular weight growth factor. When tissue was homogenized in ammonium sulfate at pH 4.5 without protease inhibitors a smaller, 16,600 dalton, growth factor was isolated. Both growth factors reacted with antisera against synthetic peptides whose sequences corresponded to the amino-terminal (1-12), Internal (33-43) and carboxyl-terminal (135-145) portions of basic fibroblast growth factor (bFGF). This suggested that the smaller growth factor was not a truncated form of (1-146) bFGF and that the larger growth factor may contain additional sequences. Amino-terminal sequencing showed the larger growth factor to have the sequence: Ala-Ala-Gly-Ser-Ile-Thr-Thr-Leu-Pro-Ala-Leu-Pro-Glu-Asp-Gly-Gly-Ser-Gly- Ala-Phe-Pro-. These results show that the larger growth factor is an 8 amino acid extended from of (1-146) bFGF and it is likely that the smaller growth factor is a proteolytic cleavage product of the larger growth factor produced during the extraction procedure.

Amino Acid Sequence↗

Mitogenic effects of hydroxyapatite and calcium pyrophosphate dihydrate crystals on cultured mammalian cells.

Synthetic hydroxyapatite (HA) crystals in 1% serum stimulated 3H thymidine uptake into quiescent canine synovial fibroblasts and human foreskin fibroblast cultures, as did 10% serum. The onset of stimulation and peak uptake of thymidine after crystal addition were delayed by 2-3 hours as compared with the effects produced by 10% serum. Stimulation of 3H thymidine uptake was proportional to the serum concentration used. HA crystals (50 micrograms/ml) stimulated nuclide uptake at each serum concentration used. 3H thymidine uptake was also proportional to the dose of HA or calcium pyrophosphate dihydrate crystals, although larger doses of the latter crystal were required to produce equivalent effects. Not all particulates were effective mitogenic agents. Latex beads and diamond crystals had no effect. Monosodium urate crystals modestly stimulated and calcium urate crystals markedly stimulated nuclide uptake. The more complex crystals found in a naturally occurring condition (calcinosis) were as mitogenic as the pure synthetic HA. The synovial cell hyperplasia sometimes associated with crystals might be explained in part by their mitogenic activity.

Animals↗