Alkaline Phosphatase Activity In Tissues Research Articles

Dynamics of serum and tissue alkaline phosphatase activity after hepatectomy in normal and cirrhotic rat liver

To investigate the pathogenesis of bile stasis after partial hepatectomy in the cirrhotic liver, the activity of alkaline phosphatase in serum and liver tissue, as well as morphologic changes in the bile canaliculi, were compared between cirrhotic and normal rats. In the normal rats, and cirrhotic rats produced by intraperitoneal thioacetamide injection, blood and liver specimens were obtained before and 2, 7, and 14 days after 70% partial hepatectomy. Serum and liver tissue alkaline phosphatase (ALPase) activity, liver weight, and electron histochemical changes in the bile canaliculi and microvilli were assessed. The normal rats showed rapid regeneration and marked elevation in the serum hepatic and tissue ALPase with a peak on day 2. This was associated with the maximal bile canalicular dilation and increased pericanalicular lipid vesicles, but without the microvilli destruction. These changes were normalized by 14 days after hepatectomy. In contrast, in the cirrhotic rats, regeneration and elevation in the serum and tissue ALPase were delayed, and higher total bile acidemia persisted. The bile canaliculi with ectasia before hepatectomy underwent no further dilation, and was associated with marked destruction of the microvilli and weakened ALPase in the bile canalicular membrane. The pericanalicular lipid vesicles was sparse. Persistent cholestasis in the remnant cirrhotic liver after hepatectomy is closely linked to the morphologic derangement and ALPase activity in the intra- and peri-bile canalicular area, and serum ALPase is useful as an index monitoring bile secretory capacity of the remnant liver.

Stimulatory effect of zinc acexamate administration on fracture healing of the femoral-diaphyseal tissues in rats

The effect of zinc acexamate on fracture healing of the femoral-diaphyseal tissues in rats was investigated in vivo. Zinc acexamate (0.3 and 10.0 mg Zn/100 g body weight per day) was orally administered to rats (4 weeks old) surgically fractured the femoral diaphysis for 14 to 28 days. Calcium content and alkaline phosphatase activity in the femoral-diaphyseal tissues were significantly decreased in rats with fracture healing, while bone acid phosphatase activity and protein content were markedly increased. The administration of zinc acexamate (10.0 mg Zn/100 g) for 28 days caused a significant increase in calcium content, alkaline and acid phosphatases activities, protein and deoxyribonucleic acid (DNA) contents in the femoral-diaphyseal tissues of rats with fracture healing. With the lower dose (3.0 mg Zn/100 g), zinc compound had a partial effect on bone components. Femoral mineral density in rats with fracture healing was significantly increased by the administration of zinc acexamate (10.0 mg Zn/100 g) for 28 days. Femoral-diaphyseal zinc content was significantly decreased in rats with fracture healing. This decrease was completely restored by the administration of zinc acexamate (10.0 mg Zn/100 g) for 28 days. The present study suggests that the supplement of zinc compound stimulates fracture healing of the femoral-diaphyseal tissues in rats.

Anabolic Effect of Wasabi Leafstalk(Wasabia japonica MATSUM.) Extract on Bone Metabolism in Rats.

The effect of wasabi leafstalk (Wasabia japonica MATSUM.) extract on bone metabolism in vivo was investigated. The extract was obtained from a homogenate with 20% ethanol and was orally administered 7 times at 24-h intervals (1.0, 10, and 40 mg/100 g body weight) to young rats. Administration of wasabi extract (1.0 and 10 mg/100 g) caused a significant increase in alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in the femoral-metaphyseal tissues; the DNA content was significantly elevated by the dose of 40 mg/100 g. In the femoral-diaphyseal tissues, alkaline phosphatase activity and calcium content were significantly increased by the administration of 1.0 and 10 mg/100 g, while the DNA content was not appreciably altered. The present study demonstrates that wasabi leafstalk extract has an anabolic effect on bone metabolism in rats in vivo.

Characterization of eleven novel mutations (M45L, R119H, 544delG, G145V, H154Y, C184Y, D289V, 862+5A, 1172delC, R411X, E459K) in the tissue-nonspecific alkaline phosphatase (TNSALP) gene in patients with severe hypophosphatasia. Mutations in brief no. 217. Online.

Hypophosphatasia is a rare inherited disorder characterized by defective bone mineralization and deficiency of serum and tissue liver/ bone/kidney tissue alkaline phosphatase (L/B/K ALP) activity. We report the characterization of tissue-nonspecific alkaline phosphatase (TNSALP) gene mutations in a series of 9 families affected by severe hypophosphatasia. Fourteen distinct mutations were found, 3 of which were previously reported in the North American or Japanese populations. Seven of the 11 new mutations were missense mutations (M45L, R119H, G145V, C184Y and H154Y, D289V, E459K), the four others were 2 single nucleotide deletions (544delG and 1172delC), a mutation affecting donor splice site (862 + 5A) and a nonsense mutation (R411X).

Potent Effect of Zinc Acexamate on Bone Components in the Femoral–Metaphyseal Tissues of Elderly Female Rats

1. The effect of zinc compounds on bone components in the femoral–metaphyseal tissues from elderly female rats (50 weeks old) was investigated in vitro. Bone tissues were cultured for 24 hr in Dulbecco's modified Eagle medium containing either vehicle or zinc compounds (10 −7 to 10 −5 M). 2. Zinc content, alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in the metaphyseal tissues were significantly increased by the presence of zinc sulfate (10 −6 and 10 −5 M), β-alanyl- l-histidinato zinc (AHZ; 10 −6 and 10 −5 M) and zinc acexamate (10 −7 to 10 −5 M). At 10 −5 M, the effect of zinc acexamate on the increase of bone components was more potent than that of zinc sulfate or AHZ. 3. The effect of zinc acexamate (10 −5 M) on the increase of alkaline phosphatase activity in the metaphyseal tissues was remarkable as compared with that of insulin (10 −8 M), estrogen (10 −9 M), insulin-like growth factor-I (10 −8 M), transforming growth factor-β (10 −10 M), sodium fluoride (10 −3 M), dexamethasone (10 −7 M) and vitamin K 2 (menaquinone-4; 10 −5 M) with an effective concentration. 4. The stimulatory effect of zinc acexamate (10 −5 M) on alkaline phosphatase activity and calcium content in the metaphyseal tissues was completely blocked by the presence of dipicolinate (10 −3 M), a chelator of zinc ion, and of cycloheximide (10 −6 M), an inhibitor of protein synthesis. 5. The present study demonstrates that zinc acexamate has a potent anabolic effect on bone components in the femoral–metaphyseal tissues from female elderly rats in vitro. The effect of zinc acexamate may be based in part on protein synthesis related to zinc ion in bone cells.

Anabolic effect of genistein and genistin on bone metabolism in the femoral-metaphyseal tissues of elderly rats: the genistein effect is enhanced by zinc.

The effect of genistein and genistin on bone components in the femoral-metaphyseal tissues obtained from elderly female rats was investigated in vitro. The metaphyseal tissues were cultured for 24 h in a medium containing either vehicle, genistein (10(-8)-10(-5) M) or genistin (10(-7)-10(-5) M). The presence of genistein or genistin caused a significant increase in alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in the metaphyseal tissues. The effect of genistein was greater than that of genistin. The bone components increased by genistein (10(-5) M) or genistin (10(-5) M) were completely blocked by the presence of cycloheximide (10(-6) M). The presence of zinc sulfate (10(-5) M) caused a significant increase in the genistein (10(-5) M)-elevated alkaline phosphatase activity, DNA and calcium contents. The enhancement with zinc was not seen by genistin (10(-5) M). The stimulatory effect of zinc on the genistein-induced increase in bone components of the metaphyseal tissues was completely blocked by the presence of cycloheximide (10(-6) M). The present results suggest that genistein and genistin have an anabolic effect on bone metabolism in the femoral-metaphyseal tissues of elderly rats, and that the genistein effect is enhanced by zinc, an essential trace element.

Anabolic effect of genistein on bone metabolism in the femoral-metaphyseal tissues of elderly rats is inhibited by the anti-estrogen tamoxifen.

Whether the anabolic effect of genistein on bone metabolism in the femoral-metaphyseal tissues of elderly rats is inhibited by the anti-estrogen tamoxifen was investigated in vitro. Bone tissues were cultured for 24 h in the medium containing either vehicle or genistein. The presence of genistein (10(-6) and 10(-5) M) caused a significant increase in calcium content, alkaline phosphatase activity and DNA content in the bone tissues. The effect of genistein (10(-5) M) in increasing bone alkaline phosphatase activity and DNA content was equal to the stimulatory effect of 17 beta-estradiol (10(-10) and 10(-9) M) on the bone components. The genistein effect was not significantly enhanced by the presence of 17 beta-estradiol (10(-10) and 10(-9) M). The anti-estrogen tamoxifen (10(-7) M) completely blocked the effect of 17 beta-estrogen in increasing bone alkaline phosphatase activity and DNA content. The genistein (10(-5) M)-induced increases in bone alkaline phosphatase activity and DNA content were completely abolished by the presence of tamoxifen (10(-7) and 10(-6) M), although tamoxifen itself had no effect on the bone components. The present study demonstrated that the anabolic effect of genistein on bone components is inhibited by anti-estrogen tamoxifen. The effect of genistein may be mediated through estrogen-like action.

A new approach using tissue alkaline phosphatase activity to identify early testicular cancer.

To establish the specificity and sensitivity of alkaline phosphatase activity in testicular tumours for the early detection of testicular germ cell tumours (GCTs). Samples from patients with GCTs of the testis were sectioned and stained using the nitroblue tetrazolium salt (NBT)/5 bromo-4-chloro-3-indolylphosphate (BCIP) Sigma Fast substrate technique (referred to as MEDC) and peroxidase antiperoxidase (PAP) assay. Additionally, several cell lines from GCTs and other tumours were cultured using standard techniques and stained using both methods, and lysates from all samples were stained after gel electrophoresis. MEDC showed strong and specific staining of testicular tumours only. The activity on gel electrophoresis appeared as two bands, one at 75 and one at 95 kDa, which was a unique combination for testicular tumours and testis cell lines. The MEDC method has extreme sensitivity and intensity for testicular tumour tissue and has potential use as a powerful tool for the early detection of testicular GCTs by analysing semen samples, particularly in patients with equivocal findings on testicular ultrasonography.

Effect of vitamin K2 (menaquinone-7) on bone metabolism in the femoral-metaphyseal tissues of normal and skeletal-unloaded rats: enhancement with zinc.

The effect of vitamin K2 (menaquinone-7) on bone metabolism in the femoral-metaphyseal tissues of normal and skeletal-unloaded rats was investigated. Skeletal unloading was designed using a model of hindlimb suspension; the rats were fed for the 4 days of unloading. The metaphyseal tissues obtained from normal and skeletal-unloaded rats were cultured for 48 h in medium containing either vehicle or vitamin K2 (10(-6) and 10(-5) M). The presence of vitamin K2 (10(-5) M) caused a significant increase in alkaline phosphatase activity and calcium content in the metaphyseal tissues from normal rats. Such an effect was not seen in the bone tissues from skeletal-unloaded rats. Additionally, the presence of zinc sulfate (10(-5) M) in effective concentration produced a significant increase in alkaline phosphatase activity and calcium content in the metaphyseal tissues from normal and skeletal-unloaded rats. In the presence of vitamin K2 (10(-5) M), the stimulatory effect of zinc sulfate on bone calcium content was appreciably enhanced; although this effect was completely abolished by cycloheximide (10(-6) M), an inhibitor of protein synthesis. This study demonstrates that the effect of vitamin K2 (menaquinone-7) on trabecular bone calcification in rats with skeletal unloading-induced osteopenia is enhanced by zinc in vitro. The enhancement with zinc may be based on a newly synthesized protein in the bone tissues.

Zinc decrease and bone metabolism in the femoral-metaphyseal tissues of rats with skeletal unloading.

Whether the decrease of zinc content in the femoral-metaphyseal tissues of rats with skeletal unloading is involved in the alteration of bone metabolism was investigated. Skeletal unloading was designed using the model of hindlimb suspension in rats. Animals were fed for 4 days with the unloading. The metaphyseal zinc content were significantly decreased by the unloading. Zinc accumulation in the metaphyseal tissues by a single oral administration of zinc sulfate (20 mg Zn/100 g body weight) was partially depressed by the unloading, although serum zinc concentration was higher than that in normal rats, suggesting an impaired movement of zinc from serum into bone tissues by the unloading. Skeletal unloading caused a significant decrease of alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the metaphyseal tissues. These decreases were completely restored by addition of zinc sulfate (10(-4) M) or beta-alanyl-L-histidinato zinc (AHZ; 10(-5) M) in a culture medium with the metaphyseal tissues in vitro. The effects of zinc compounds were abolished by the presence of cycloheximide (10(-8) M), suggesting that the zinc effect is based on a newly synthesized protein. Dipicolinate (10(-4) and 10(-5) M), a potent zinc-chelating agent, caused an appreciable decrease of zinc content and alkaline phosphatase activity in the metaphyseal tissues. This decrease was restored by zinc supplement. The present results suggest that the skeletal unloading-induced decrease of zinc content in the femoral-metaphyseal tissues plays a role in the deterioration of bone metabolism in the unloaded rats.

Effect of insulin administration on bone formation is impaired in rats with skeletal unloading.

The effect of insulin administration on bone formation was investigated in the femoral diaphysis of rats with skeletal unloading. When the femoral-diaphyseal tissues from normal rats were cultured in the presence of insulin (10(-9) and 10(-8) M) for 48 h, the hormone produced a significant increase of alkaline phosphatase activity, collagen, deoxyribonucleic acid (DNA) and calcium contents in the bone tissues, indicating that it has a direct stimulatory effect on bone formation. Moreover, when normal rats received a subcutaneous administration of insulin (5.0 IU/100 g body weight) 5 times at 24-h intervals, the hormone caused a significant increase of alkaline phosphatase activity, DNA and calcium contents in the femoral diaphysis. Those increases were not seen by insulin administration to the skeletal-unloading rats with hindlimb hang for 4 d. The present results indicate that the stimulatory effect of insulin on bone formation is lost by skeletal unloading. The finding further supports the view that insulin impairment plays an important role in the deterioration of bone formation by microgravity.

Toxicity of the Latex of Euphorbiales. Effect on Acid and Alkaline Phosphatases of the SnailLymnaea acuminata

ABSTRACT The latex of four plants belonging to the family Euphorbiaceae viz. Jatropha gossypifolia, Euphorbia royleana, Euphorbia antisyphlitica and Euphorbia lactea cristata caused significant reductions in the acid and alkaline phosphatase activity in nervous tissue of the snail Lymnaea acuminata. The reduction in the activity of both the phosphatases was time and dose dependent; the order of effectiveness was as follows: Euphorbia lactea cristata > Euphorbia antisyphlitica > Euphorbia royleana > Jatropha gossypifolia. It has been suggested that the inhibition of these enzymes might be through activation of Protein Kinase C.

Simulated weightlessness and bone metabolism: impairment of insulin effect on alkaline phosphatase activity in bone tissue

The effect of simulated weightlessness on bone alkaline phosphatase was investigated after skeletal unloading for up to 4 days. The skeletal unloading was designed by using the model of hindlimb hang in rats. The femoral-diaphyseal fragments obtained from rats bred with skeletal unloading were cultured for 24 h at 37 degrees C in 5% CO2/95% air in Dulbecco's Modified Eagle Medium (high glucose). The bone alkaline and acid phosphatase activity were significantly decreased by skeletal unloading. When the bone tissue was cultured with synthetic [Asu1,7] eel calcitonin (3 and 30 nM), the hormone caused a significant increase of alkaline phosphatase activity in the bone tissues from rats with normal and skeletal-unloading. In culture with insulin (1.0 and 10 nM), skeletal unloading impaired the effect on insulin to increase bone alkaline phosphatase activity. Meanwhile, the culture with zinc sulfate (10 and 100 microM), which can increase bone protein synthesis, caused a remarkable elevation of alkaline phosphatase activity in the bone tissues form rats with normal and skeletal-unloading. Insulin (10 nM) did not alter the zinc effect. These findings suggest that the skeletal unloading with hindlimb hang causes the impairment of insulin's effect to increase alkaline phosphatase activity in the femoral diaphysis of rats, although the effects of calcitonin and zinc were not altered.

Ultrastructural localisation of alkaline phosphatase in the intertubular tissue of the testis in the domestic fowl

Alkaline phosphatase activity in the intertubular tissue of the testes of the domestic fowl was examined using an ultracytochemical technique based on the lead capture method. In the interstitial tissue, the Leydig cells, transitional cells and the fibroblasts displayed enzyme activity on their cell membranes. Vacuoles located in the transitional cells were lined by reaction products of enzyme activity, whereas the vacuoles representing extracted lipid droplets and present mainly in the Leydig cells were free of enzyme activity. In the peritubular tissue the cell processes of fibroblasts showed enzyme activity on the cell membranes and in pinocytotic vesicles. Cell processes lying adjacent to blood vessels showed pronounced activity. In the blood vessel itself some activity was present in the basement membrane and the endothelium. The surface of the red blood cell showed moderate activity. The possible role of alkaline phosphatase in the transfer of hormone from the Leydig cells to the seminiferous tubules and from the seminiferous tubules to the interstitium is discussed. The myoid cells and their processes were devoid of enzyme activity.

Intestinal Infection with Nematospiroides Dubius Selectively Increases Lactase Expression by Mouse Jejunal Enterocytes

1. Intestinal structure, lactase (beta-galactosidase; EC 3.2.1.23) activity and alkaline phosphatase activity have been determined in mouse jejunal and ileal tissues before and during infection with the intestinal parasite Nematospiroides dubius. 2. Oral infection with small numbers of N. dubius larvae caused villus height, crypt depth and enterocyte migration rate to increase in the mouse jejunum. None of these effects occurred in ileal tissue. 3. Lactase activity also increased in jejunal, but not ileal, tissue of infected mice. This increase was associated with a doubling of the rate at which activity appeared in the brush-border membrane of enterocytes during migration over the basal regions of jejunal villi. Alkaline phosphatase activity in jejunal tissue remained unchanged in infected mice. 4. Attention is drawn to the fact that this is the first occasion when crypt cell hyperplasia has been found to be positively correlated with an increase in lactase activity and a decrease in cytotoxic/suppressor T-cells. Further work is needed to establish the primary cause of these effects.

Enzyme histochemical demonstration of alkaline phosphatase activity in plastic-embedded tissues using a Gomori-based cerium-DAB technique.

We describe a new method for light microscopic demonstration of alkaline phosphatase (ALP) activity in plastic-embedded sections. Rat tissues were fixed in acetone (-20 degrees C), infiltrated in glycol methacrylate (GMA), and embedded at 0 degrees C. Sections were cut at 1 and 2 microns, dried at room temperature, and incubated in the conventional Gomori medium. Cerium chloride was used to convert calcium phosphate into cerium phosphate, which was subsequently converted into cerium perhydroxide. The slight yellow precipitate of cerium perhydroxide was amplified using 3,3'-diaminobenzidine tetrahydrochloride (DAB). For comparison, tissue sections were processed according to the calcium-cobalt method. The method described combines exact localization of ALP activity with optimal preservation of tissue morphology.

Comparison of alkaline phosphatase activity in normal, lactating and neoplastic rat mammary tissue

1. 1. Alkaline phosphatase activity in NMU-induced rat mammary tumours was compared with activity in normal and lactating mammary gland. 2. 2. Both tumour and normal mammary alkaline phosphatase were sensitive to heat inactivation and inhibition by phenylalanine. 3. 3. Specific activity of enzyme in tumours was comparable to normal mammary tissue. 4. 4. Mammary gland alkaline phosphatase increased markedly in late pregnancy and early lactation. 5. 5. Bromocryptine treatment had no effect on enzyme activity in lactating mammary gland.

Tissue calcification and alkaline phosphatase activity in uraemic rats.

The activity and localization of alkaline phosphatase activity (AP) in aorta and heart, and the incidence of calcifications in aorta, heart and kidney as well as cardial fibrosis were studied in uraemic rats treated with 1,25-dihydroxycholecalciferol (1,25-DCHH) and Nifedipine. 1,25-DHCC treatment elevated the serum Ca x P product and aggravated the development of renal and aortic calcifications and cardial fibrosis. Nifedipine did not protect against calcifications, but decreased the incidence of cardial fibrosis. The activity of AP was increased in the thoracic aorta in uraemia independent of 1,25-DHCC or Nifedipine treatment or presence of calcification. No changes of the AP activity were found in the heart.

Alkaline Phosphatase Activities Following Repeated Suramin Administration in Some Rat Tissues in Relation to Their Functions

The effect of repeated administration of suramin, a trypanocide and filariacide, on the level of alkaline phosphatase activity in some rat tissues and organs was investigated. Daily administration of this drug to rats resulted in a very significant increase in enzyme activity in the kidney and small intestine immediately after the first dose. Activity observed in the liver was not affected until after the third dose when the level was increased. In the heart and the large intestine, administration of the drug did not affect the enzyme activity levels throughout the duration of drug administration. These results suggest that repeated suramin administration may result in very large increases in alkaline phosphatase activity in the tissues and organs which are involved in active transport mechanisms.

Distribution of 20 enzymes in the midgut region of the earthworm, Lumbricus terrestris L., with particular emphasis on the physiological role of the chloragog tissue

1. 1. Lumbricus terrestris midgut was separated into peripheral chloragocytes, peripheral intestinal epithelium and typhlosole, and the distributions of 20 enzymes were assayed. 2. 2. The peripheral chloragog tissue contained ⩽4% of the midgut dehydrogenase activities, but about 44% or more of the total catalase, Superoxide dismutase, β- d-glucuronidase, alkaline phosphatase, esterase, (δ-aminolevulinate dehydratase and porphyrin synthetase activities. 3. 3. The midgut contained about 70% or more of the total midbody isocitrate dehydrogenase, glutaminate dehydrogenase and NADH- and NADPH-diaphorase activities. 4. 4. Most of the intermediary metabolism, including amino acid deamination, probably takes place in the midgut epithelium, which is also the site of the mixed function oxidase system. 5. 5. The chloragog tissue probably synthesizes the blood hemoglobin and is responsible for the destruction of Superoxide and hydrogen peroxide from the interaction between blood hemoglobin and molecular oxygen. 6. 6. The high alkaline phosphatase activity in ehloragog tissue may reflect transmembrane transport activity or may be involved in glycogen break-down. 7. 7. Neither xanthine oxidase nor uric acid, xanthine or hypoxanthine were present in detectable amounts.