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Query: EC:3.1.3.1 (
alkaline phosphatase
)
47,916
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Many human cancer cell lines which have been maintained in fetal bovine serum (FBS)-supplemented medium produce and secrete many substances such as
transferrin
, alpha 1-antitrypsin, alpha 2-macroglobulin,
alkaline phosphatase
, gamma-glutamyltranspeptidase, creatine kinase, carcinoembryonic antigen, alpha-fetoprotein, carbohydrate antigen 19/9, and cytokines including colony-stimulating factors and transforming growth factor, and further they may produce small amounts of unknown substances. Usually, small amounts of substances have to be concentrated as highly as possible for detection, but FBS interferes with this procedure. A protein-free culture system is an ideal method for detecting small quantities of substances which originate from cancer cells without interference by FBS. However, we were concerned that protein-free culture may interrupt the production of the substances which have been produced in FBS-supplemented medium. In this study, we investigated the productibility of 46 kinds of well-known substances in ten newly established cell lines derived from human pancreatic cancer. These cell lines were propagated in a protein-free non-FBS-supplemented medium. Of the ten cases, one cell line alone that was derived from acinal cell carcinoma propagated as a semisuspension; on the other hand, nine cell lines that were derived from ductal cell carcinoma propagated as monolayers without piling up. This method prolongs the doubling time, which is not affected by the addition of FBS. The spent media of these cell lines were collected aseptically after the removal of cell debris and concentrated by ultrafiltration using a Pericon cassette followed by lyophilization. Using 46 kinds of available antibodies, we investigated whether or not the substances which react to these antibodies could be detected in the spent media and in the cells by enzyme-linked immunosorbent assay, Western blot analysis, and immunocytochemistry. Among these cell lines, HPC-Y11 produced and secreted the most kinds of substances, and the production of those substances was lowest in HPC-Y0. In conclusion, our protein-free culture system can be available in every laboratory, since this is not only an economical method, but also an effective method for the saving of purification procedures. Moreover, this is a most suitable method for surveying unknown substances derived from cancer cell lines.
...
PMID:Characterization of new human pancreatic cancer cell lines which propagate in a protein-free chemically defined medium. 220 67
Biochemical polymorphism of 116 dwarf goats has been studied by starch gel electrophoresis. The systems haemoglobin, ceruloplasmin and erythrocyte esterase were monomorphic. 4 loci were found to be polymorphic: amylase,
transferrin
,
alkaline phosphatase
, and albumin. The allele frequency of these traits was given.
...
PMID:[Biochemical polymorphism in a dwarf goat population]. 226 70
A series of proteins (albumin,
transferrin
, alpha 1-antitrypsin, alpha-fetoprotein and pancreatic oncofetal antigen) and enzymes (gamma-glutamyltranspeptidase, aminopeptidase M,
alkaline phosphatase
, alpha-glucosidase and protease) was measured in fetal meconium extracts. There were 19 fetuses thought to have cystic fibrosis (CF), 13 with neural tube defects, three with chromosome abnormalities and 19 normal controls, all with gestational ages between 18 and 21 weeks. With the exception of alpha-fetoprotein, all the proteins and enzymes were significantly elevated in the CF meconium extracts. The most definitive indicator of a CF fetus was the albumin concentration, where the mean level was five times that found in the control groups. However, five of 19 fetuses assumed to have CF had albumin in the normal range. In these cases the meconium protease levels were grossly elevated. Furthermore, in the same five fetuses meconium concentration of pancreatic oncofetal antigen, a protein synthesized in the fetal pancreas, was also greatly raised. We suggest that post-mortem examination of a fetus thought to have CF should include measurement of meconium albumin, protease and pancreatic oncofetal antigen.
...
PMID:Biochemical analysis of meconium in fetuses presumed to have cystic fibrosis. 242 27
The end-stage maturation of neutrophilic granulocyte precursor cells isolated from normal human bone marrow by Ficoll density centrifugation was studied in a liquid culture assay system used previously to study the maturation of guinea pig granulocyte precursors. Dialyzed normal human serum induced end-stage morphological maturation of human granulocyte precursors and this induction was proportional to a serum level of up to 5.0% in the assay medium. At serum concentrations greater than 5.0% a pronounced inhibition of maturation was observed. Passage of serum through a DEAE-Fractogel 650S column equilibrated with 0.01 M phosphate buffer (pH 7.0) resulted in the binding of the end-stage granulocyte maturation factor to the column. The activity eluted from the column in a fraction containing 17% of the starting serum protein that was inhibitor-free and was also capable of inducing the appearance of granulocyte
alkaline phosphatase
, a specific biochemical marker for granulocyte end-stage maturation. GMF is most likely a protein since it was destroyed by protease digestion. The data also indicate that neither purified human
transferrin
nor human recombinant granulocyte colony-stimulating factor can substitute for human serum GMF as a granulocyte end-stage maturation factor in this assay system. It was observed, however, that purified human
transferrin
greatly potentiated the effect of GMF suggesting that
transferrin
plays a supporting role in the end-stage maturation of human granulocytes in vitro. To our knowledge the evidence presented here indicates for the first time the existence of a neutrophilic granulocyte end-stage maturation factor in normal human serum.
...
PMID:Evidence for a factor in normal human serum that induces human neutrophilic granulocyte end-stage maturation in vitro. 247 45
Proximal tubules were prepared from rat kidney cortex by collagenase digestion and purified by Percoll gradient centrifugation. Their enrichment was estimated by comparing the specific activities of various cell-specific enzymes in homogenates of renal cortex and of the isolated tubules. The tubules were cultured in a 50:50 mixture of Dulbecco's modified Eagle's and Ham's F12 media supplemented with insulin,
transferrin
, epidermal growth factor, hydrocortisone, and prostaglandin E1. After 2 to 3 d an extensive outgrowth of epithelial cells developed from the attached tubules. After 5 to 7 d near confluent monolayers were obtained. Hormonal responsiveness, marker enzyme activities, and transport properties were determined to further characterize the primary cultures. The cultured cells exhibited increased cyclic AMP production in response to parathyroid hormone but not calcitonin or vasopressin, consistent with the absence of cells derived from distal and collecting tubules. The cells also retained significant levels of 25-hydroxyvitamin D3-1 alpha-hydroxylase,
alkaline phosphatase
, and gamma-glytamyl-transpeptidase, three enzymes that are primarily associated with the proximal tubule. The cultured epithelial cells also exhibit a Na+-dependent phosphate and glucose transport systems. Therefore, the cells retain many functional properties that are characteristic of proximal tubules. Thus, the primary cultures should be suitable for the study of processes that occur specifically within this segment of the rat nephron.
...
PMID:Characterization of primary cell cultures derived from rat renal proximal tubules. 254 89
The effect of 17 beta-estradiol (E) on an osteoblast-like cell line, UMR106, was studied in vitro. The concentrations of
transferrin
and seven enzymes (gamma glutamyl transferase,
alkaline phosphatase
, acid phosphatase, lactate dehydrogenase, creatine kinase, alanine aminotransferase and aspartate aminotransferase) were measured in these cells after incubation in culture medium containing either E or the vehicle. E treatment increased five of the seven enzymes and increased the
transferrin
concentration in the UMR106 cells while simultaneously reducing the proliferation rates. 4-Hydroxytamoxifen, an estrogen antagonist, produced a mild estrogen agonist action on growth rates and enzyme concentrations in the UMR106 cells. When E was present simultaneously, the agonist properties of 4-hydroxytamoxifen were enhanced. These studies show that E enhanced activity of five enzymes and the
transferrin
content of UMR106 cells after a 2-day incubation. 4-Hydroxytamoxifen enhanced the E effect, illustrating that estrogen antagonists may manifest agonist or antagonist properties depending on the model. These results extend our previous observations showing a direct effect of E in vitro on osteoblast-like cells.
...
PMID:Biochemical effects of 17 beta-estradiol on UMR106 cells. 256 66
Postmenopausal women lose bone mineral density and this loss can be prevented by estrogen administration. Although the skeletal effects of estrogens have been regarded previously as indirect, estrogen receptors have been discovered in cultured human osteoblasts and related cell lines. The UMR106 cell line derived from a rat osteogenic osteosarcoma is such an osteoblast model. We have shown direct effects of estradiol (E) on these cells in vitro, inhibiting growth and stimulating
alkaline phosphatase
activity (AP) corrected for cell number. This response was maximal at E conc. of 10(-10) M in serum and Phenol Red free medium, was metabolite specific and cell cycle-dependent. These cells contain high affinity binding sites with a Kd of 0.5 nM. Estrogen receptors were detected by the monoclonal antibody H-222 on Western blot after initial immunoprecipitation to concentrate the proteins. E treatment increased several enzymes including creatine kinase and LDH isoenzymes along with increments in intracellular
transferrin
. Transforming growth factor-beta is secreted by these cells. Secretion of this peptide was stimulated by E. TGF-beta mediated the transient growth inhibition associated with E treatment. Insulin like growth factors (IGF) are also secreted by these cells with IGF-II concentrations in the culture medium being eight times higher than IGF-I levels. E treatment increased the concentrations of both IGFs in the culture medium after a 3 day incubation. Exposure of E treated cells manifested a mitogenic response and reduced AP, indicating that E induced receptors for IGFs. These findings establish direct effects of E on osteoblastic cells in vitro and demonstrate responses to E at many levels. These osteoblast responses in vitro suggest an important role for sex steroids in the development and function of the osteoblast lineage.
...
PMID:Estrogens and the skeleton: cellular and molecular mechanisms. 262 18
When
transferrin
receptors of human erythroleukemic cells were pulse-labeled with [35S]methionine and then chased in the absence of radioactive precursor, the first detectable immunoprecipitable form of the receptor had a molecular mass of 85 kDa. This form of the receptor was converted to the mature form of 93 kDa with a half-time of about 40-60 min. Both the immature (85 kDa) and mature (93 kDa) receptors associated as dimers, the native form of the receptor. The 85-kDa, as well as the 93-kDa, receptors bound to a monoclonal antibody raised against the transferrin receptor or to
transferrin
-Sepharose. In order to determine whether glycosylation was necessary for ligand binding, purified receptors were isolated from cells grown in the presence of tunicamycin. When K562 cells were grown in the presence of tunicamycin, an 80-kDa nonglycosylated form of the receptor was synthesized. This nonglycosylated receptor was also capable of dimer formation; however, much less of it reached the cell surface than the fully glycosylated form, although both untreated and tunicamycin-grown cells appeared to synthesize
transferrin
receptors at similar rates. Although the number of receptor molecules/cell was similar in control and tunicamycin-treated cells, the nonglycosylated receptors exhibited a much lower affinity for
transferrin
than those of untreated cells; in contrast, when receptors were purified by immunoprecipitation and digested with bacterial
alkaline phosphatase
, no difference was observed between the affinity of these receptors and undigested immunoprecipitated receptors. These results suggest that glycosylation is not necessary for specific binding of
transferrin
to its receptor, but the affinity of this binding can be influenced greatly by the presence or absence of carbohydrate residues.
...
PMID:Changes in glycosylation alter the affinity of the human transferrin receptor for its ligand. 272 54
The Ascanian multi-foetus and pure-bred Karakul sheep reared in the steppe region of the Ukraine are characterized by five-allelic status of
transferrin
and by diallelic serum arylesterase and
alkaline phosphatase
. Besides the basic HbA and HbB alleles the rare HbC type has been revealed in the multi-foetus Karakul. Reliable differences in concentrations of the
transferrin
and haemoglobin alleles between the multi-foetus Karakul population and that of the pure-bred Karakul have been found.
...
PMID:[Comparative analysis of the gene pool of the Ascanian polycarpous and purebred Karakul' sheep populations]. 274 96
Growth characteristics and the expression of trophoblast-associated markers by six cell lines generated from midgestation chorioallantoic placentas of outbred (Holtzman) and inbred (Lewis, PVG.RT Ir8) rats were evaluated. The cells comprising all cell lines were epithelioid (contained cytokeratin-type intermediate filaments), had normal (2n, 4n) DNA content, and synthesized the extracellular matrix glycoprotein laminin. Variability was observed among the lines in all other characteristics: median cell size, rate of growth, serum dependency, responses to
transferrin
and dibutyryl cyclic adenosine-3',5'-monophosphate, synthesis of some major proteins,
alkaline phosphatase
activity, and the expression of immunoreactive placental lactogen-II. In general, cell lines with smaller mean cell sizes grew rapidly and required little serum for maintenance in vitro; cell lines with larger mean sizes grew more slowly and preferred higher concentrations of serum. Some associations between mean cell size/rate of growth and other characteristics were observed. No major differences were apparent between cell lines generated from outbred and inbred rat placentas. Trophoblast cell lines expressing distinct phenotypes provide a valuable new approach for studying a wide range of trophoblast cell activities.
...
PMID:Isolation of phenotypically distinct trophoblast cell lines from normal rat chorioallantoic placentas. 278 30
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