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1.
J Pharm Sci ; 89(3): 417-27, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10707021

ABSTRACT

The molecular mobility of amorphous pharmaceutical materials is known to be a key factor in determining their stability, reactivity, and physicochemical properties. Usually such molecular mobility is quantified using relaxation time constants. Typically relaxation processes in amorphous systems are non-exponential and relaxation time constants are usually obtained from experimental data using a curve fitting procedure involving the empirical Kohlrausch-Williams-Watts (KWW) equation. In this article we explore the possible relationship between the KWW curve fitting parameters (tau(KWW), beta(KWW)) and common statistical measures of the average and the distribution (e.g., median, standard deviation) of the relaxation time values. This analysis is performed for several common statistical distributions (e.g., normal, lognormal, and Lorentzian), and the results are compared and analyzed in the context of pharmaceutical product stability predictions. The KWW function is able to describe relaxation processes stemming from several different statistical distribution functions. Under some circumstances the "average" relaxation time constant of the KWW equation (tau(KWW)) is significantly different from common statistical measures of the central value of a distribution (e.g., median). Simply knowing the relaxation time constants from the fit of the KWW equation is not sufficient to completely characterize and quantify the molecular mobility of amorphous pharmaceutical materials. An appreciation of the distribution of relaxation times and the resulting effects upon the KWW constants should be considered to be essential when working with amorphous pharmaceutical materials, especially when attempting to use relaxation time constants for predicting their physical or chemical stability.


Subject(s)
Chemistry, Pharmaceutical/statistics & numerical data , Pharmaceutical Preparations/chemistry , Algorithms , Chemical Phenomena , Chemistry, Physical , Crystallization , Drug Stability , Models, Statistical
2.
Pharm Res ; 16(5): 672-5, 1999 May.
Article in English | MEDLINE | ID: mdl-10350009

ABSTRACT

PURPOSE: To evaluate thermomechanical analysis (TMA) as a technique for determining the viscosity of amorphous pharmaceutical materials. This property of amorphous drugs and excipients is related to their average rate of molecular mobility and thus to their physical and chemical stability. METHODS: Indomethacin was selected as a model amorphous drug whose viscosity has previously been reported in the literature. A Seiko TMA 120C thermomechanical analyzer was utilized in isothermal penetration mode to determine the viscosity of the amorphous drug over the maximum possible range of temperatures. RESULTS: Using a cylindrical penetration geometry it was possible to accurately determine the viscosity of amorphous indomethacin samples by TMA over the temperature range from 35 to 75 degrees C. The results were consistent with those reported in the literature using a controlled strain rheometer over the range 44-75 degrees C. The limiting lower experimental temperature for the TMA technique was extended to significantly below the calorimetric glass transition temperature (Tg approximately 42 degrees C), thus allowing a direct experimental determination of the viscosity at Tg to be made. CONCLUSIONS: Thermomechanical analysis can be used to accurately determine the viscosity of amorphous pharmaceutical materials at temperatures near and above their calorimetric glass transition temperatures.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/chemistry , Calorimetry/methods , Indomethacin/chemistry , Glass , Rheology , Temperature , Viscosity
3.
Nucl Med Biol ; 23(1): 69-73, 1996 Jan.
Article in English | MEDLINE | ID: mdl-9004917

ABSTRACT

Z-17 alpha-iodovinyl-11 beta-chloromethyl-estradiol (Z-CMIV), a new selective estradiol derivative, can easily be labeled with high efficiency by radioactive iodine isotopes. Biodistribution studies and quantitative scintigraphic imaging of human breast carcinoma xenografts in mice demonstrated continuous and selective accumulation of the [123I]Z-CMIV, in estrogen receptor (ER)-positive target tumors, with significantly high target/nontarget ratio up to 48 h post-injection. A receptor-mediated mechanism of concentration of Z-CMIV in target tissues was confirmed by scintigraphic imaging and by biodistribution studies.


Subject(s)
Adenocarcinoma/diagnostic imaging , Breast Neoplasms/diagnostic imaging , Estradiol/analogs & derivatives , Receptors, Estrogen/metabolism , Adenocarcinoma/metabolism , Animals , Breast Neoplasms/metabolism , Estradiol/pharmacokinetics , Female , Humans , Iodine Radioisotopes , Isotope Labeling , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation/diagnostic imaging , Ovariectomy , Radionuclide Imaging , Tissue Distribution
4.
Calcif Tissue Int ; 54(4): 304-11, 1994 Apr.
Article in English | MEDLINE | ID: mdl-8062145

ABSTRACT

Ligated ileal loops, 30 cm in length, of 4-month-old male Wistar rats were instilled with 3 ml of a 10 mM CaCl2 solution (added with 0.25 muCi 45Ca) in the absence (control) or presence of 100mM sorbitol, L-xylose, or creatine. Ileal calcium (Ca) transport, measured by plasma 45Ca appearance, was found to be similar 30 minutes after fluid instillation in all four instances. However, thereafter, 45Ca appearance in plasma did not increase further in control animals whereas it increased twice as much during the subsequent 30 minutes in the presence of sorbitol, L-xylose, or creatine. However, when loops of similar length were instilled with only 1.0 ml of such solutions, the sorbitol effect was already observed during the first 30 minutes. The stimulation of ileal Ca absorption induced by the presence of sorbitol appeared to be due to a cellular effect, associated with a decreased flux across the paracellular pathway, as indicated by 3H-mannitol absorption. The presence of sorbitol in instilled ileal solution induced a significant decrease in luminal Na, K, bicarbonate, and Cl concentrations at each time point studied (30, 60, 120, or 240 minutes after instillation). Thirty minutes after instillation, no difference in soluble Ca concentration was observed between control and experimental rats. After 60 minutes, Ca concentration was dramatically decreased in control rats but it remained nearly constant in experimental animals. Thus, the presence of substances enhancing ileal Ca transport favored the maintenance of soluble Ca in ileal solution during longer time periods than their absence. In the ileal enterocyte, these substances induced a twofold increase of ATP content compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Calcium/metabolism , Creatine/pharmacology , Ileum/metabolism , Intestinal Mucosa/metabolism , Sorbitol/pharmacology , Xylose/pharmacology , Absorption/drug effects , Adenosine Triphosphate/metabolism , Animals , Bicarbonates/metabolism , Chlorine/metabolism , Energy Metabolism , Ileum/drug effects , Ileum/ultrastructure , Intestinal Mucosa/cytology , Intestinal Mucosa/ultrastructure , Male , Mannitol/metabolism , Mitochondria/drug effects , Mitochondria/ultrastructure , Phosphates/metabolism , Potassium/metabolism , Rats , Rats, Wistar , Sodium/metabolism
5.
Calcif Tissue Int ; 50(5): 433-8, 1992 May.
Article in English | MEDLINE | ID: mdl-1317741

ABSTRACT

In the femoral extremities of the adult rat containing the metaphysis, the epiphyseal cartilage, and the epiphysis, four alkaline phosphatase (AP) forms were distinguished on polyacrylamide gel electrophoresis. Two soluble forms were present in the 160,000 g supernatant: one of Mr 165 kDa and another of Mr 110-115 kDa, which exhibited a strong catalytical activity. Moreover, from the pellet, three membrane-bound forms of Mr 130, 110-115, and 100 kDa could be extacted with sodium deoxycholate. When denaturated AP was visualized by postelectrophoretic autoradiography of the phosphorylated intermediates, subunits always appeared as three monomers of Mr 75-80, 60-70, and 50-60 kDa. As four native forms but only three types of subunits were found to be present in the femur, it seems that, apart from homodimers, some heterodimers could also occur. Three types of diets were administered to three groups of rats for 5 weeks. Two are known to disturb bone mineralization: (1) a vitamin D3-deficient diet, and (2) the same as (1) but enriched with 12% sorbitol. The third was a normal diet containing vitamin D3. Concerning the effects on AP of dietary sorbitol and the vitamin D3-deficient diet, it was found that rats receiving the diet supplemented with sorbitol showed a substantial rise in the activity of the Mr 165 kDa form with the concomitant appearance of a new monomer of Mr 100 kDa. In contrast, rats fed the vitamin D3-deficient diet always displayed an increase in enzyme activity, principally of the Mr 100 and 110 kDa forms.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Alkaline Phosphatase/analysis , Cholecalciferol/pharmacology , Femur/enzymology , Food, Formulated/analysis , Isoenzymes/analysis , Sorbitol/pharmacology , Alkaline Phosphatase/chemistry , Alkaline Phosphatase/physiology , Animals , Bone Density/drug effects , Cholecalciferol/administration & dosage , Cholecalciferol/analysis , Electrophoresis, Polyacrylamide Gel , Growth Plate/enzymology , Male , Osteoporosis/etiology , Rats , Rats, Inbred Strains , Rickets/etiology , Sorbitol/administration & dosage , Sorbitol/analysis , Vitamin D Deficiency/enzymology
6.
C R Acad Sci III ; 315(1): 31-5, 1992.
Article in French | MEDLINE | ID: mdl-1422917

ABSTRACT

Several compounds, either metabolizable such as sorbitol and creatine or unmetabolizable such as L-xylose or ethyl-ethanolamine, increase ileal calcium transport, similarly to the well documented effect of lactose. These compounds increase the duration time but not the rate of calcium transport. They have no specific effect on intestinal calcium absorption but they prolong the presence of soluble calcium in the ileal loop, thus allowing the calcium absorption. This stimulating effect of these compounds on mineral absorption is associated with an increase of ileal mucosal ATP content.


Subject(s)
Calcium/pharmacokinetics , Creatine/pharmacology , Ileum/metabolism , Phosphates/metabolism , Sorbitol/pharmacology , Animals , Biological Transport/drug effects , Ethanolamines/pharmacology , Intestinal Absorption , Rats , Xylose/pharmacology
7.
Enzyme ; 46(6): 276-83, 1992.
Article in English | MEDLINE | ID: mdl-1308851

ABSTRACT

A kinetic study of the inhibition of several alkaline phosphatase (AP isoenzyme activities by phenobarbital was carried out using p-nitrophenylphosphate (10 mM) as a substrate at pH 9.8 in a 300-mM Hepes buffer. AP from bovine kidney, calf intestine, bovine liver, and rat bone was used. Over a phenobarbital concentration range of 20-400 mM, all these isoenzymes were inhibited in an uncompetitive manner with a Ki of 200 mM for intestinal AP, and in a linear mixed-type manner for all the other isoenzymes tested. The Ki values were 10, 40 and 55 mM for kidney, bone and liver AP, respectively. The use of 15 mM carbonate-bicarbonate or 400 mM diethanolamine buffer did not modify the degree of inhibition of intestinal AP activity. Dixon plots of the reciprocal of reaction velocity versus inhibitor concentration either at different substrate concentration or at different DEA concentration indicate uncompetitive inhibition for the intestinal enzyme. This in vitro inhibitory effect of phenobarbital is in contrast to its in vivo stimulating action on AP. However, in the whole animal, the effects of phenobarbital administration probably represent the sum of multiple effects.


Subject(s)
Alkaline Phosphatase/antagonists & inhibitors , Bone and Bones/enzymology , Intestines/enzymology , Isoenzymes/antagonists & inhibitors , Kidney/enzymology , Liver/enzymology , Phenobarbital/pharmacology , Alkaline Phosphatase/isolation & purification , Animals , Cattle , Ethanolamines/pharmacology , Isoenzymes/isolation & purification , Kinetics , Substrate Specificity
8.
Miner Electrolyte Metab ; 18(1): 61-8, 1992.
Article in English | MEDLINE | ID: mdl-1328835

ABSTRACT

Calcium transport in the ileal-ligated loop was studied in the adult rat in the presence of either phosphate alone or phosphate-binding compounds, namely either hydroxylated or aminated substances. Sorbitol or creatine (50 mM) added to a 10-mM CaCl2 solution, which was instilled into ileal loop, markedly enhanced calcium transport, as determined by 45Ca radioactivity appearing in plasma and from 45Ca radioactivity disappearing from the loop. The presence of both compounds maintained Ca soluble in an instilled solution at a constant concentration, whereas with a control solution the Ca concentration progressively decreased towards zero after an incubation period of 60 min. Phosphate, which was either simultaneously added with sorbitol or creatine or which was present as sorbitol or creatine phosphate, led to an equally marked decrease in calcium transport. Calcium transfer was even more reduced when phosphate alone was present with calcium in the ileal loop, in the absence of sorbitol. Similar to the above phosphate-binding compounds, adenosine and its constitutive component, ribose, increased calcium transfer, whereas adenine, the other constitutive component of adenosine, was ineffective. Guanosine was twice more active than adenosine in stimulating ileal calcium transport. Interestingly, the structure of guanosine allows the binding of two phosphates, with one binding site being on the ribose and the other on the guanine base moiety. Thus guanosine is capable of binding a greater amount of phosphate than the two other aminated compounds examined, namely adenosine and alanine, when transphosphorylation from ATP is studied with intestinal microvilli preparations.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Calcium/metabolism , Carrier Proteins/metabolism , Ileum/metabolism , Phosphates/metabolism , Adenosine/pharmacology , Adenosine Triphosphate/metabolism , Animals , Biological Transport/drug effects , Calcium Radioisotopes , Guanosine/pharmacology , Male , Microvilli/enzymology , Phosphate-Binding Proteins , Phosphocreatine/pharmacology , Phosphotransferases/metabolism , Rats , Rats, Wistar , Solubility , Sorbitol/pharmacology
9.
Reprod Nutr Dev ; 32(5-6): 453-60, 1992.
Article in English | MEDLINE | ID: mdl-1292482

ABSTRACT

The effect of calcium, phosphate and the sugars lactose and sorbitol on the intestinal absorption of manganese were studied in adult male rats. Gastric gavage showed that lactose (100 mM or 200 mM) increased the hepatic retention of 54Mn, while phosphate decreased it. In situ ileal loop studies indicated that Mn absorption was normally complete in 30 min. Sorbitol had no effect on uptake during this period, but extended Mn absorption from 30 min to 120 min. Low concentrations of Mn (10 microM) did not alter the enhancing effect of lactose on calcium transport (10 mM), but the enhancing effect of lactose on Mn transport was blocked by this high calcium concentration. Intestinal alkaline phosphatase activity was rapidly stimulated by Mn. These similarities plus the competition between cations, especially calcium, suggest that a common mechanism exists in their intestinal transport.


Subject(s)
Calcium/metabolism , Intestinal Absorption/drug effects , Manganese/metabolism , Alkaline Phosphatase/metabolism , Animals , Binding, Competitive , Calcium/pharmacology , Cations, Divalent , Kinetics , Lactose/pharmacology , Liver/drug effects , Liver/metabolism , Male , Manganese/pharmacology , Phosphates/pharmacology , Rats , Rats, Wistar , Sodium Chloride/pharmacology , Sorbitol/pharmacology
10.
Int J Biochem ; 23(2): 175-80, 1991.
Article in English | MEDLINE | ID: mdl-1900247

ABSTRACT

1. There is a good correlation between the capacity of sugars to stimulate calcium transfer and their capacity to be phosphorylated by the intestinal alkaline phosphate with a part of the phosphate liberated from an ester phosphate. 2. On the sugar dependent and sugar independent calcium transfer, inhibitors of this enzyme act differently. 3. Phosphate, a competitive inhibitor suppresses both transfers. 4. Only the dependent sugar transfer was suppressed with phloridzin acting competitively at the sugar site, or with EDTA which could react close to the active site. 5. L-phenylalanine and phenobarbital, not competitive inhibitors does not act on either type of calcium transfer, the sugar dependent or the sugar independent.


Subject(s)
Alkaline Phosphatase/antagonists & inhibitors , Calcium/metabolism , Intestinal Absorption/drug effects , Animals , Binding Sites , Binding, Competitive , Carbohydrates/pharmacology , Duodenum/metabolism , Edetic Acid/pharmacology , Ileum/metabolism , Male , Phenobarbital/pharmacology , Phenylalanine/pharmacology , Phlorhizin/pharmacology , Phosphates/pharmacology , Phosphorylation , Rats , Rats, Inbred Strains
11.
Arch Int Physiol Biochim ; 98(4): 141-8, 1990 Aug.
Article in English | MEDLINE | ID: mdl-1707609

ABSTRACT

For four weeks after weaning, rats were fed either on a diet without any calcium utilization factors (-D) or on the same diet with cholecalciferol (+D) or sorbitol (S). In the -D group, blood calcium levels decreased whilst alkaline phosphatase activities in blood and bone were increased. For +D and S groups, these parameters were normal. Using everted or in situ ligatured loops, calcium transfer from a CaCl2 + 45Ca solution was measured in the duodenum, the jejunum and in the ileum. Alkaline phosphatase activity from these regions was also measured. For the three diets and for all regions of the intestine, there was a good correlation between calcium transfer and phosphatase activity. These values were higher in the duodenum than in the ileum or jejunum, and also higher in the ileum in the +D group than in the -D and S groups although this was not significant. These low levels in the S group which were, sometimes, even lower than those seen in the -D group contrasted with blood and bone levels of alkaline phosphatase, which were normal for the S and +D groups. There was also a discrepancy between the low values found for both phosphatase activity and calcium transfer in rats S in the experiments where the calcium transfer assay was carried out in calcium solution and those found in experiments were both calcium and carbohydrate were present. In the latter, enhanced levels of intestinal phosphatase activity were observed, as well as a marked, albeit delayed, increase in intestinal calcium transfer. Onset latency and rapid offset are reminiscent of induction of bacterial enzymes by carbohydrates.


Subject(s)
Alkaline Phosphatase/metabolism , Calcium/metabolism , Cholecalciferol/pharmacology , Diet , Intestine, Small/metabolism , Sorbitol/pharmacology , Alkaline Phosphatase/isolation & purification , Animals , Biological Transport/drug effects , Bone and Bones/metabolism , Calcium/blood , Dietary Carbohydrates/pharmacology , In Vitro Techniques , Intestinal Absorption/drug effects , Intestine, Small/enzymology , Male , Rats , Rats, Inbred Strains
12.
Acta Biochim Pol ; 37(4): 445-50, 1990.
Article in English | MEDLINE | ID: mdl-2100896

ABSTRACT

Four alkaline phosphatase forms from adult rat femur were distinguished on polyacrylamide gel electrophoresis: two soluble forms of Mr 165,000 and 110,000 in the water extract, and three membrane-bound forms of Mr 130,000, 110,000 and 100,000 extractable with deoxycholate. Alkaline phosphatase after SDS-treatment disintegrated into three kinds of monomers: of Mr 80,000, 65,000 and 50,000. The soluble fraction (extract I) contained subunits of Mr 80,000 and 55,000--whereas the pellet fraction (extract II), subunits of Mr 65,000 and 50,000. Since for native forms only three types of subunits were found it seems that, apart from homodimers, there are also some heterodimers composed of the Mr 65,000 and 50,000 subunits forming the native enzyme of Mr 110,000-115,000. Two denatured monomers: of Mr 80,000 and 50,000 may form two native homodimeric forms of Mr 165,000 and 100,000 while in the pellet two monomers: of Mr 65,000 and 50,000 may correspond to three native alkaline phosphatase forms: of Mr 130,000, 110,000-115,000 and 100,000. Probably the Mr 110,000-115,000 form is a heterodimer composed of subunits of Mr 65,000 and 50,000.


Subject(s)
Alkaline Phosphatase/chemistry , Bone and Bones/enzymology , Alkaline Phosphatase/metabolism , Animals , Autoradiography , Calcification, Physiologic/physiology , Chemical Fractionation , Electrophoresis, Polyacrylamide Gel , Femur , Immunoenzyme Techniques , Male , Molecular Weight , Rats , Rats, Inbred Strains , Serum Albumin, Bovine
13.
Acta Biochim Pol ; 37(4): 451-6, 1990.
Article in English | MEDLINE | ID: mdl-2100897

ABSTRACT

The effect of vitamin D3-deficiency and dietary sorbitol on serum calcium level, the activity and alkaline phosphatase (AP) pattern in femoral epiphysis were studied. Rats fed a diet supplemented with sorbitol or vitamin D3 showed the same serum calcium concentration and AP activity in serum and femur. Rats fed a vitamin D3-deficient diet displayed decreased serum calcium concentration and increased AP activity both in serum and femur. Four forms of AP were isolated from the femur of these rat groups: of Mr 100,000, 110,000, 130,000 and 165,000. Rats receiving the diet supplemented with sorbitol showed a marked rise in the activity of the Mr 165,000 form, and appearance of a new monomer of 100,000, never formed in two remaining groups.


Subject(s)
Alkaline Phosphatase/metabolism , Epiphyses/enzymology , Sorbitol/administration & dosage , Vitamin D Deficiency/enzymology , Alkaline Phosphatase/analysis , Alkaline Phosphatase/chemistry , Animals , Autoradiography , Calcium/blood , Electrophoresis, Polyacrylamide Gel , Femur , Male , Molecular Weight , Rats , Rats, Inbred Strains , Sorbitol/pharmacology
14.
Biol Cell ; 69(3): 179-89, 1990.
Article in English | MEDLINE | ID: mdl-2097003

ABSTRACT

The effects of vitamin D on the intramuscular distribution of total and bound calcium, phosphate and on available cytosolic calcium, were investigated in skeletal muscle. Total calcium and phosphorus were measured on ashed subcellular fractions of muscles from vitamin D-repleted and vitamin D-deprived rats. The variations in available calcium were followed by determining the activities of calcium-sensitive enzymes in isolated cytosol. Bound-calcium was revealed ultra-microscopically by pyroantimonate. In vitamin D-repleted muscles, the pyroantimonate method revealed specific areas of intense bound-calcium deposition: the myofibrils, where they formed pronounced lines parallel to the Z-bands. In vitamin D-deficient muscles, the calcium-pyroantimonate deposits appeared clearly reduced. This loss was accompanied by a marked reduction in total calcium and phosphorus in all the subcellular fractions, as compared to vitamin D-repleted muscles. Unexpectedly, the activity of the Ca(+)-activated isocitrate-dehydrogenase was increased in the cytosol, while that of the Ca2(+)-inhibited pyruvate-kinase decreased. Prolonged vitamin D-administration to vitamin D-repleted rats led to an intensification of calcium-pyroantimonate deposits and a general increase in total calcium and phosphorus, but no change in the cytosolic Ca2(+)-sensitive enzyme activities. Cessation of vitamin D-administration to vitamin D-repleted rats produced a regression of calcium-pyroantimonate deposits, a general decrease of total calcium and phosphate levels, and stimulation of the Ca2(+)-activated isocitrate-dehydrogenase accompanied by lowering of the Ca2(+)-inhibited pyruvate-kinase. The results clearly indicate a correlation between vitamin D-repletion and the total and bound calcium content of skeletal muscle. In addition, they demonstrate an apparent contradiction between the decrease of total and bound calcium, and the activities of cytosolic Ca2+ sensitive enzymes during vitamin D-deprivation, which can only be explained by an increase in available calcium. It is suggested that vitamin D stimulates intramuscular mechanisms tending to lower available calcium by inactivating the cation via the formation of calcium chelates.


Subject(s)
Calcium/metabolism , Muscles/metabolism , Vitamin D/pharmacology , Aconitate Hydratase/metabolism , Animals , Antimony/pharmacokinetics , Calcium/administration & dosage , Calcium/pharmacokinetics , Cytosol/enzymology , Isocitrate Dehydrogenase/metabolism , Male , Muscle Proteins/metabolism , Phosphorus/metabolism , Pyruvate Kinase/antagonists & inhibitors , Rats , Rats, Inbred Strains , Vitamin D/administration & dosage , Vitamin D Deficiency/metabolism
15.
Acta Biochim Pol ; 37(1): 177-80, 1990.
Article in English | MEDLINE | ID: mdl-2087908

ABSTRACT

The influence of ethanol on formation of phosphorylated intermediates of different forms of alkaline phosphates in brush border of adult rat was studied. Ethanol enhanced phosphorylation of the Mr 65,000 subunit of jejunal enzyme but had no effect on the Mr 90,000 subunit. Commercial highly purified alkaline phosphate in the presence of ethanol also displayed a two times higher phosphorylation ability.


Subject(s)
Alkaline Phosphatase/metabolism , Ethanol/pharmacology , Jejunum/enzymology , Microvilli/enzymology , Muscle, Smooth/enzymology , Animals , Cattle , Kinetics , Macromolecular Substances , Molecular Weight , Phenobarbital/pharmacology , Phosphorylation , Rats , Rats, Inbred Strains
16.
Acta Biochim Pol ; 37(1): 191-6, 1990.
Article in English | MEDLINE | ID: mdl-2087910

ABSTRACT

The effect of vitamin D3 on the activity of different forms of alkaline phosphatase as well as on the ability of their subunits to form phosphorylated intermediates in the presence of [gamma-32P]ATP was studied. In experiments with rats fed a diet enriched in vitamin D3 phosphatase F3 activity was doubled and the radioactivity of the phosphorylated intermediate of the F3 subunit was greatly enhanced. In contrast, in the case of phosphatase F1 the enzymic activity and the radioactivity of F1 subunit phosphorylated intermediate remained unchanged. In rats fed a vitamin D3-deficient diet, the phosphatase F3 activity was greatly reduced and there were only traces of radioactivity of F3 subunit phosphorylated intermediate.


Subject(s)
Alkaline Phosphatase/metabolism , Calcitriol/pharmacology , Intestine, Small/enzymology , Isoenzymes/metabolism , Microvilli/enzymology , Vitamin D Deficiency/enzymology , Adenosine Triphosphate/metabolism , Animals , Duodenum/enzymology , Ileum/enzymology , Jejunum/enzymology , Macromolecular Substances , Male , Molecular Weight , Phosphorylation , Rats , Rats, Inbred Strains , Reference Values
17.
Pathol Biol (Paris) ; 36(6): 819-24, 1988 Jun.
Article in French | MEDLINE | ID: mdl-3138625

ABSTRACT

Adults rats receive by gavage 10 mM CaCl2 [+45Ca] solution containing or not 100 mM glucid, 10-100 mM EDTA or these both compounds. Calcium transfer is determined by the evaluation of [45Ca] in intestine and feces as well as in plasma and femur. Basic Ca++ transfer which corresponds to the CaCl2 solution was doubled in the presence of glucids. EDTA addition abolishes completely the glucid effect, exercising any influence on basic CA++ transfer. Injected into ligaturated ileal loop, the glucid gives the same effect. But the addition of phosphate not only removed glucid action but also inhibits the basic Ca++ transfer. The glucids, known acceptors of phosphate, increase Ca transfer. EDTA and phosphate, alkaline phosphatase inhibitors, exhibit an opposite effect. Phlorizin, as it was seen previously, acts exactly as EDTA. All these facts question: the simultaneous transfer of Ca and glucid, the possibility of a glucid phosphorylation, the part in these events of alkaline phosphatase, phosphorylating, phosphatasic and phosphorylable microvillar protein.


Subject(s)
Alkaline Phosphatase/antagonists & inhibitors , Calcium/metabolism , Intestinal Absorption/drug effects , Alkaline Phosphatase/pharmacology , Animals , Calcium/administration & dosage , Carbohydrates/pharmacology , Edetic Acid/administration & dosage , Edetic Acid/pharmacology , Male , Rats , Rats, Inbred Strains
18.
Life Sci ; 43(25): 2059-65, 1988.
Article in English | MEDLINE | ID: mdl-3210900

ABSTRACT

Three forms of alkaline phosphatase have been isolated from different sections of the small intestine: F3 180 kDa from the duodenum; F2 150 kDa principally jejunal; F1 120 kDa the only ileal form. Their catalytic properties have been compared as well as the electrophoretic properties the dimer and monomer of their phosphorylated intermediates. Pi was a competitive inhibitor of F1 and F3, whereas glycerophosphate was competitive inhibitor only of F3. Pi was a non competitive inhibitor of F2 and of a mixture F1 plus F3. Heating the phosphorylated enzyme preparations led to their dissociation into the phosphorylated monomers: F1 and F3 appear to be homodimers 65 kDa and 90 kDa peptides respectively whilst F2 seems to be a dimer formed from one of each monomer. F1 was phosphorylated faster but less intensively than F3. F2 was strongly phosphorylated over a long time-course and its 65 kDa monomer fraction was phosphorylated more strongly for longer than that from F1.


Subject(s)
Alkaline Phosphatase/isolation & purification , Intestine, Small/enzymology , Isoenzymes/isolation & purification , Alkaline Phosphatase/metabolism , Animals , Duodenum/enzymology , Ileum/enzymology , Isoenzymes/metabolism , Jejunum/enzymology , Kinetics , Macromolecular Substances , Molecular Weight , Organ Specificity , Rats , Rats, Inbred Strains
19.
C R Acad Sci III ; 307(10): 585-90, 1988.
Article in French | MEDLINE | ID: mdl-3142642

ABSTRACT

Semi-purified preparations were obtained from duodenal, jejunal or ileal mucosa containing one of the three alkaline phosphatase forms. The Mr of the isoenzymes were for F1 120, F2 150, F3 180 kDa. F1 was the only species found in the ileum; F2 was duodenal but mainly jejunal; F3 was found mainly in duodenum. These enzymes forms were the only phosphorylable proteins in these preparations. Following treatment with denaturing agents they were separated on gel electrophoresis into monomers F': F'1 65, F'2 65 and 90, F'3 90 kDa. Thus F2 could be an heterodimer. All were far more phosphorylated from ATP than from inorganic phosphate. As compared with F1, F3 was relatively more sensitive to ATP and less sensitive to inorganic phosphate.


Subject(s)
Alkaline Phosphatase/metabolism , Intestine, Small/enzymology , Animals , Duodenum/enzymology , Ileum/enzymology , Intestinal Mucosa/enzymology , Jejunum/enzymology , Phosphorylation , Rats , Rats, Inbred Strains
20.
Reprod Nutr Dev (1980) ; 27(3): 641-8, 1987.
Article in French | MEDLINE | ID: mdl-3616125

ABSTRACT

At alkaline or neutral pH and with PNPP or ATP as substrates, the effect of cations on alkaline phosphatase of jejunal microvilli was shown to differ, i.e. Mg++ had an activating effect at high concentrations, Zn++ slightly inhibiting, Ca++ and Co++ were inactive at all concentrations. At neutral pH with PNPP as a substrate, Mg++ and Co++ were active, Ca++ inactive and Zn++ slightly inhibiting at high concentrations. Using ATP as a substrate, all four cations were activators at all concentrations studied (0.2 to 5 mM). In the latter physiological conditions (pH and substrate), similar effects of the cations were observed with microvilli from other segments of rat small intestine.


Subject(s)
Alkaline Phosphatase/metabolism , Jejunum/enzymology , Metals/pharmacology , Adenosine Triphosphate/metabolism , Animals , Calcium/pharmacology , Cations, Divalent/pharmacology , Cobalt/pharmacology , Hydrogen-Ion Concentration , Jejunum/drug effects , Magnesium/pharmacology , Microvilli/drug effects , Microvilli/enzymology , Nitrophenols/metabolism , Organophosphorus Compounds/metabolism , Rats , Rats, Inbred Strains , Zinc/pharmacology
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