Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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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)

Testicular peritubular and prostatic stromal cells produce extracellular matrix elements and paracrine factors that modulate the cytodifferentiation and function of the corresponding epithelial cells. The present paper describes the establishment and characterization of five rat testicular cell lines with peritubular characteristics and one prostatic stromal cell line. Four peritubular cell lines were isolated after transfection of a mixed peritubular-Sertoli cell culture with a v-myc-containing plasmid. The same immortalization procedure applied to prostatic stromal cells yielded one cell line. An additional testicular cell line arose by spontaneous immortalization during serial subculture. Except for one testicular cell line (RTC-8T1), the morphology of all of the immortalized cell lines strongly resembled that of primary cultures of peritubular and stromal cells. Flow cytometric analysis demonstrated that all cell lines scored positive for alpha-smooth muscle isoactin and negative for cytokeratins, confirming their myofibroblast-like nature. None of the cell lines, however, stained positive for alkaline phosphatase, and androgen receptor expression was also lost. Typical Leydig cell characteristics, such as steroidogenesis, and Sertoli cell markers, such as transferrin secretion, were absent. Coculture of the cell lines with Sertoli cells resulted in the formation of tubular structures. A cell attachment assay and an enzyme-linked immunosorbent assay for fibronectin confirmed the production of extracellular matrix elements by all of the established cell lines. Media conditioned by the cell lines stimulated Sertoli cell transferrin production. The active principle was partially purified and resembled the P-MOD-S-like factors produced by primary cultures of peritubular and stromal cells. It is concluded that the immortalized cell lines have retained several of the characteristics of primary cultures of peritubular and stromal cells and may be useful for further studies on mesenchymal-epithelial interactions in testis and prostate.
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PMID:Characterization of newly established testicular peritubular and prostatic stromal cell lines: potential use in the study of mesenchymal-epithelial interactions. 778 11

Caco-2 cell human colon adenocarcinoma cell line was used to study the hormonal regulation of small intestinal epithelial cell differentiation. We had previously shown that insulin-transferrin-selenium and triiodothyronine (5 x 10(-8) M)-supplemented medium can best replace serum after 2 days of culture for both the maintenance and differentiation of Caco-2 cells. The present study demonstrates that precoating petri dishes with complete serum allows the growth and differentiation of Caco-2 cells seeded directly in serum-free medium. On the other hand, precoating with dialyzed serum inhibits alkaline phosphatase and dipeptidyl-dipeptidase IV activities by more than 50%. The results obtained with complete serum-precoated culture plates indicate that there is no synergy between insulin and triiodothyronine because cells maintained in transferrin-selenium and triiodothyronine-supplemented medium, with or without insulin, express comparable enzyme activities. Moreover, large increases in alkaline phosphatase and dipeptidyl-dipeptidase IV activities were observed when triiodothyronine was added to the culture medium by the time confluency was reached. In contrast, gamma-glutamyltransferase was lowered to a greater extent when triiodothyronine was present from the beginning of culture. These findings show that triiodothyronine preferentially stimulates alkaline phosphatase and dipeptidyl-dipeptidase IV activities during the differentiation period whereas it selectively inhibits gamma-glutamyltransferase during the proliferation phase. Triiodothyronine acts in a dose-dependent manner.
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PMID:Alkaline phosphatase and peptidase activities in Caco-2 cells: differential response to triiodothyronine. 788 29

Human fetal kidney explants can be maintained during 5 days in Leibovitz's L15, a basic serum-free medium. Because culture conditions are minimal for growth and differentiation, DNA synthesis drastically decreases during the first 48 h, but stabilizes thereafter. The addition of insulin plus transferrin significantly restores this important cellular function in kidneys of fetuses younger than 16 wk. However, renal explants from older fetuses are more difficult to culture: they respond less to growth factors and are more prone to necrosis. The objective of this study was to verify the influence of tetracycline, an antibiotic with anti-collagenase potential, on cultured kidney explants aged 17 to 20 wk. The addition of 20 micrograms/ml tetracycline did not influence DNA synthesis nor the effectiveness of insulin plus transferrin on cell proliferation. Nor did it change the activities of alkaline phosphatase and gamma-glutamyltransferase, two enzymic markers of brush border differentiation. After 5 days in L15 alone, explants often showed necrosis and an important reduction in both weight and volume. Insulin plus transferrin significantly restored these parameters to control values observed at Day 0, but evidence of necrosis was still present. Tetracycline alone markedly reduced explant necrosis resulting in a significant increase in weight and volume. The effectiveness of insulin plus transferrin on explant morphometry was not improved when tetracycline was added as third factor. These results indicate that insulin plus transferrin restores explant mass through cell proliferation, whereas tetracycline does so possibly through a reduction in extracellular matrix degradation. The two effects are not additive in cultured mid-term fetal kidneys.
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PMID:Positive influence of tetracycline on human fetal kidney in serum-free organ culture. 791 74

Genetic polymorphisms of hemoglobin, transferrin, amylase, alkaline phosphatase, and ceruloplasmin, interior blood indices, and the reproductive function of servicing bulls were studied. Correlations between the heterozygosity level with respect to blood protein systems and enzymes and interior blood indices and the reproductive function of servicing bulls were detected.
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PMID:[The interrelation of the interior indices of sires with the level of the heterozygosity for the polymorphic blood protein and enzyme systems]. 794 Oct 19

The modulation of transferrin secretion by FSH and epidermal growth factor (EGF) was studied in highly pure, primary cultures of immature rat Sertoli cells grown on a reconstituted basement membrane (Matrigel) in bicameral chambers. Sertoli cell purity was assessed by (1) morphometry, (2) alkaline phosphatase cytochemistry (a specific marker enzyme for peritubular cells) and (3) immunocytochemistry for the alpha-isoform of smooth muscle actin in contaminating peritubular cells. Results revealed a less than 0.5% peritubular cell contamination. During initial periods of culture with EGF or FSH alone or in combination, both EGF and FSH alone maintained transferrin secretion over basal values and their effects were additive. At subsequent times, EGF alone maintained transferrin secretion, but to less extent than did FSH alone, and inhibited significantly the ability of FSH to maintain transferrin secretion. The ratio of polarized transferrin secretion in response to FSH, EGF, or in combination was also examined. FSH significantly reversed the polarity of transferrin secretion, whereas EGF, although significantly reducing the ratio of apical to basal transferrin secretion, did not lead to a preferential basal secretion of transferrin. The change in the apical:basal transferrin secretion ratio, however, was not due to a reversal of the apically secreted transferrin towards a basal direction, but rather to an increase in the total basally secreted transferrin. The effects of cell density effects on transferrin secretion were then examined. At low cell density, the relative ability of EGF and FSH together to maintain transferrin secretion was greater than at high cell density, but overall transferrin secretion was greater as cell density increased. The inhibition of FSH by EGF on transferrin secretion was also density dependent: EGF significantly inhibited FSH effects at low cell density, but failed to do so at high cell density. These results suggest that regulation of transferrin secretion by Sertoli cells appears to be under dual control, involving both FSH and EGF and may provide an explanation for the mechanism by which EGF exerts a regulatory role in spermatogenesis.
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PMID:Modulation of transferrin secretion by epidermal growth factor in immature rat Sertoli cells in vitro. 802 75

Short-term cultures of purified murine trophoblast were used to investigate the potential trophic effects of a number of cytokines. Both granulocyte-macrophage colony stimulating factor (GM-CSF) and colony stimulating factor-1 (CSF-1) increased [3H]thymidine (TdR) uptake (3-8 times control values) by trophoblast harvested from placentae on day 12 or 14 of pregnancy. In contrast, interleukin-3 (IL-3) had only a mild stimulatory effect ([3H]TdR uptake 1.5 times control), and IL-2 did not alter the level of DNA synthesis in these cells. Immunocytochemical analysis confirmed that the cells engaged in DNA synthesis were cytokeratin-positive trophoblast cells and revealed that these cells predominantly bore markers (alkaline phosphatase, transferrin receptors) characteristic of trophoblast cells from the placental labyrinth. The increased DNA synthesis observed after exposure to GM-CSF or CSF-1 was not associated with a change in the proportion of nuclei involved in synthesis, nor did it result in significantly increased trophoblast cell numbers in the cultures. These findings suggest that DNA-synthesizing trophoblast cells were not proliferating, but were more likely engaged in endoreduplicative cycles leading to the formation of terminally differentiated trophoblast giant cells. These results caution against the presumption of proliferation when measuring [3H]thymidine incorporation by placental or trophoblast cells in standard in vitro cultures. In addition, taken together with the reports of high levels of CSF-1 in the pregnant uterus and the expression of the CSF-1 receptor on placental trophoblast cells, they suggest that the hemopoietic cytokines may play a role in the differentiation and/or function of trophoblast cells in the developing murine placenta.
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PMID:GM-CSF and CSF-1 stimulate DNA synthesis but not cell proliferation in short-term cultures of mid-gestation murine trophoblast. 804 Aug 36

Urea kinetic modelling (UKM) was used to assess adequacy of dialysis in 50 CAPD patients. Nutritional status was assessed from the measurement of visceral protein status (total protein, albumin, transferrin, immunoglobulins, complement), somatic protein status (anthropometry), and dietary intake (1 week weighed dietary inventory and normalized protein catabolic rate (NPCR) from UKM). Morbidity was assessed from the peritonitis and admission history. Mean Kt/V (corrected to x3 weekly dialysis) was 0.66 +/- 0.02. Dietary protein intake estimated from the NPCR (1.08 +/- 0.03 g kg-1 day-1) correlated well (r = 0.72, P < 0.001) with that estimated from the dietary inventory (1.10 +/- 0.04 g kg-1 day-1). There was a strong correlation between Kt/V and NPCR corrected for actual weight (r = 0.65, P < 0.001), but when NPCR was corrected for IBW this correlation was weaker (r = 0.35, P < 0.05). Patients were divided by Kt/V into two groups (> 0.65, n = 22 and < 0.65, n = 28). There were no significant differences in the indices of visceral protein status between the two groups. Weight, height, BMI, fat free mass and arm muscle area were significantly greater in the group Kt/V < 0.65. Residual renal function (creatinine clearance) was higher in the group Kt/V > 0.65 (3.8 +/- 0.7 versus 1.9 +/- 0.5 1/24 h, P < 0.05) and plasma creatinine less (913 +/- 51 versus 1265 +/- 51 mumol/l, P < 0.001). Hb, potassium, bicarbonate, phosphate, alkaline phosphatase, PTH, and blood pressure were not different. Neither was there any difference between the two groups in any of the indices of morbidity.
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PMID:Adequacy of dialysis and nutritional status in CAPD. 815 6

The serum activities or concentrations of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), albumin, gamma-glutamyl transpeptidase (GGT), bilirubin (BIL), cholic acid (CHOL), chenodeoxycholic acid (CHENO), and transferrin with isoelectric point 5.7, and the urinary excretion of albumin were determined among male current or former house painters (n = 135) and house carpenters (n = 71) who had worked in their trades for at least 10 years before 1970. Workers who showed a value above the 90th percentile among the carpenters in at least one of the tests ASAT, ALAT, GGT, BIL, CHOL, or CHENO were regarded as showing "possible signs of liver dysfunction". Each participant's lifetime solvent exposure was evaluated by interview. The painters were divided into categories with low, intermediate, and heavy cumulative exposure during life (LTSE) or during the most exposed year (MEYSE). All participants stated none or slight recent exposure. The prevalence of possible signs of liver dysfunction increased with solvent exposure category according to LTSE as well as MEYSE with a numerically higher risk estimate in the heavy exposure category for MEYSE than for LTSE. ALP activity increased with exposure category according to both exposure estimates. This increase seemed to be due to an interaction between exposure to solvents and current or previous long term intake of medicines potentially toxic to the liver. None of these results was affected by whether or not the subjects had been exposed to solvents during the year before the investigation. The exposure to solvents was not significantly related to any other outcome variable. It is concluded that long term heavy exposure to solvents may elicit changes in conventional liver function tests indicative of a mild chronic effect on the liver. The findings also suggest that heavy solvent exposure during short time periods is a more likely cause of the findings than lifetime cumulative solvent exposure and that an interaction between solvent exposure and medicines potentially harmful to the liver may be important in the causation of the effects.
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PMID:Liver function tests and urinary albumin in house painters with previous heavy exposure to organic solvents. 819 87

We examined the alterations of proliferative activity and c-myc expression of a colon cancer cell line (Caco-2) during its spontaneous differentiation. Caco-2 cells were cultured in various types of media and the degree of differentiation was monitored in terms of dome formation in cell monolayers and expression of alkaline phosphatase (ALP) activity. In Caco-2 cells cultured with Eagle's minimum essential medium (EMEM) containing 10% fetal calf serum (FCS), dome formation was demonstrated and ALP activity was markedly increased after the cells reached confluence. Five-fold reduction of c-myc mRNA and a marked decrease in S-phase cells were observed in the differentiated cells. These changes were not induced in FCS-free EMEM. The addition of insulin and transferrin to FCS-free EMEM did not induce cell differentiation or reduction of c-myc mRNA expression. When Caco-2 cells were cultured with three different serum-free media, the induction of dome formation and the increase of ALP activity were observed to varying degrees. Expression of c-myc mRNA in the cells cultured with one serum-free medium decreased to a level similar to that in fully differentiated cells cultured with EMEM containing 10% FCS. These results suggest that a spontaneous switch from a proliferative state with high c-myc expression to differentiated phenotype occurs after cells reach confluence and depends on the culture conditions.
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PMID:Changes of proliferative activity and phenotypes in spontaneous differentiation of a colon cancer cell line. 839 33

Isolated proximal tubular (PT) and distal tubular (DT) cells from rat kidney were cultured for up to 9 days under serum-free, hormonally-defined conditions on 35-mm polystyrene culture dishes. Several hormonal and growth factor supplements were assessed for their ability to promote growth (increased protein and DNA content) and stability of differentiated phenotype (high activities of gamma-glutamyltransferase and alkaline phosphatase as brush-border membrane markers in PT cells; maintenance of high activities of glutamate dehydrogenase as a mitochondrial marker in both PT and DT cells; maintenance of low and high activities of lactate dehydrogenase in PT and DT cells, respectively; expression of cytokeratins). Basal supplemented media (DMEM/F12, 1:1 v/v) contained insulin, hydrocortisone, epidermal growth factor, sodium selenite and transferrin as supplements. Additionally, triiodothyronine selectively promoted growth and stability of differentiated phenotype in PT cells and thyrocalcitonin selectively promoted growth and stability of differentiated phenotype in DT cells. On Day 3 of primary culture, PT and DT cells were incubated for up to 8 h with either tert-butyl hydroperoxide (tBH; 0.5-10 mM), methyl vinyl ketone (MVK; 1-10 mM), or p-aminophenol (PAP; 1-10 mM) and cellular injury, as assessed by cellular release of lactate dehydrogenase, was determined. DT cells were significantly more susceptible to injury from both tBH and MVK, but the two cell populations were equally susceptible to injury from PAP, which is the same susceptibility pattern seen in freshly isolated cells. These results suggest that primary cultures of rat renal PT and DT cells reflect similar biochemical properties as freshly isolated cells and are, therefore, useful models for study of chemically induced injury.
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PMID:Susceptibility of primary cultures of proximal tubular and distal tubular cells from rat kidney to chemically induced toxicity. 854 48


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