Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A thirteen-year-old Dutch warmblooded mare was referred to the Faculty of Veterinary Medicine because of a sinusitis. She was thin with a potbellied appearance. Her coat was dull with long wavy hair. Unilateral (left) purulent nasal discharge was evident. A cbc revealed leucopenia (3.9 G.L.-1) and plasma biochemical analysis revealed a plasma glucose concentration of 10.1 mmol.L-1. Thermostable alkaline phosphatase (at 65 degrees C during 2 minutes) could not be demonstrated. Basal plasma cortisol concentration was lowered (114 nmol.L-1) and basal plasma ACTH concentration was highly elevated (815 pg.ml-1), indicating adrenocortical insufficiency. A dexamethasone-suppression test was performed by intramuscular administering of 10 mg of dexamethasone showing a suppression of 32% instead of at least 50%. At necropsy a pituitary pars intermedia adenoma was found.
 ...
PMID:[A horse with Cushing's disease]. 165 May 4

Non-radioactive in situ hybridization employing detection of biotin-labeled probes by an alkaline phosphatase-based procedure has proven useful for demonstrating a wide variety of mRNA species. With certain developers, the alkaline phosphatase reaction product is both light microscopically visible and fluorescent. We have exploited this to perform simple double-stainings for mRNA's and their corresponding peptide products in human insulin and gastrin cells and in rat ACTH, MSH and gastrin cells. Such stainings show that nearly all of these cells simultaneously contain both the peptide hormone and its corresponding mRNA. Human gastrin cells show a differentiated localization of gastrin mRNA and gastrin. Thus, while gastrin immunofluorescence predominates in secretory granules present in the basolateral region of the cells, gastrin mRNA is virtually restricted to the supranuclear region of the cells. Here it may be preferentially associated with granular endoplasmic reticulum. The strict subcellular localization of gastrin mRNA differs from that of general polyadenylated RNA in the G cells and raises questions whether specific transport routes or sites of accumulation for defined mRNA species exist.
 ...
PMID:Combined non-radioactive detection of peptide hormones and their mRNA's in endocrine cells. 166 Aug 59

Plasma adrenocorticotropin (ACTH) levels increase after acute cold exposure. The purpose of this study was to determine if there were parallel changes in pituitary proopiomelanocortin (POMC) mRNA. Male rats were exposed to cold (3-5 degrees C) or a novel environment for 15 or 30 min. Others were unstressed. POMC mRNAs in frozen sections or dissociated cells were hybridized with a photobiotinylated oligonucleotide probe which was detected in situ by streptavidin-alkaline phosphatase. The percentage of area labeled for POMC mRNA was quantified by the Cue-3 color image analysis system. In frozen sections, 24-fold increases in the percentage of area labeled for POMC mRNA were evident in intermediate lobes (IL) 30 min after stress. No change was seen in anterior lobes (AL). If the ALs were dissociated, a 66-99% increase in percentage of labeled cells was detected 2-3 hr after the cold exposure. Fifteen min of cold stress (CS) also caused a 117% increase in the area of label for POMC mRNA per corticotrope. No change was seen after 30 min. Exposure to a novel environment caused a 73% increase in the percentage of area labeled for POMC mRNA per AL corticotrope and an 11-fold increase in the IL. These results indicate that both AL corticotropes and IL melanotropes are stimulated by acute exposure to cold and novel environments.
 ...
PMID:Changes in rat pituitary POMC mRNA after exposure to cold or a novel environment, detected by in situ hybridization. 185 78

In order to determine the nature of afferent fibres contacting thyrotropin-releasing hormone (TRH)-synthesizing neuronal cell bodies in the rat hypothalamic paraventricular nucleus, we used dual immunostaining procedures which employed antibodies to ACTH (to label proopiomelanocortin (POMC) neurons), neuropeptide Y (NPY) and dopamine-beta-hydroxylase (D beta H) and peroxidase-labeled goat anti-rabbit IgG as a first sequence and antibodies to a cryptic fragment (Phe178-Glu199) of pro-TRH (to label TRH neurons) and alkaline phosphatase-labeled goat anti-rabbit IgG as the second sequence. A rich innervation of the paraventricular nucleus by immunoreactive POMC, NPY and D beta H fibres was observed. Numerous NPY and POMC fibres were in intimate anatomic proximity and often appeared to surround in remarkable density TRH-containing cell bodies. Less frequent appositions between D beta H fibres and TRH cell bodies were detected. These results strongly suggest that TRH neurons might be regulated by POMC, NPY as well as adrenergic and/or noradrenergic systems. These interactions might be the neuroanatomical basis for the already observed effects of opiate peptides, NPY and catecholamines on TSH secretion.
 ...
PMID:Anatomical interactions of proopiomelanocortin (POMC)-related peptides, neuropeptide Y (NPY) and dopamine beta-hydroxylase (D beta H) fibers and thyrotropin-releasing hormone (TRH) neurons in the paraventricular nucleus of rat hypothalamus. 190 63

Non-radioactive in situ hybridization using biotinylated oligodeoxynucleotides and a detection protocol involving monoclonal antibiotin antibodies and the alkaline phosphatase-anti-alkaline phosphatase system was employed for quantitation by image analysis. Calibrations of the image analysis system with neutral density filters revealed that the grey levels recorded were strongly linearly correlated to the absorbance (r2 = 0.97) in the range studied in tissue specimens (0-0.8 optical density or absorbance units). Several methodological parameters, including light source stability, section thickness, probe concentration and development time were initially optimized. Model systems revealed that the grey level measured varied linearly with the logarithm of the target concentration. Moreover, histophysiological studies on adrenalectomized and sham-operated rats documented that previous biochemical data on an 8- to 10-fold increase in anterior lobe proopiomelanocortin (POMC) mRNA levels 8 days after adrenalectomy are accounted for both by an increased ACTH cell concentration and content of POMC mRNA, as well as by increases in ACTH cell sizes and cell numbers. Also in agreement with biochemical data, image analysis did not reveal significant differences between OD's of melanotrophs in adrenalectomized and sham-operated animals. To our knowledge, these data are the first to document that non-radioactive in situ hybridization can be employed for relative quantitation. A particular advantage of this approach is the good morphological definition which permits parallel analyses of densitometric values, cell sizes and cell areas/cell numbers.
 ...
PMID:Quantitative non-radioactive in situ hybridization. Model studies and studies on pituitary proopiomelanocortin cells after adrenalectomy. 205 May 40

Rabbit adrenal 17 alpha-hydroxylase activity has previously been shown to increase dramatically following ACTH stimulation. The present study was designed to determine whether the increase in enzyme activity could be correlated with an increase in P-450(17 alpha) protein measured by immunoblotting using an anti-porcine P-450(17 alpha) antibody. It was found that the total and specific contents of rabbit adrenal immunoreactive P-450(17 alpha) were increased 6- to 8-fold and 4-fold, respectively, after ACTH stimulation. The results were similar whether the detection system was 125I-labeled protein A or an alkaline phosphatase-conjugated second antibody. Corresponding increases in 17 alpha-hydroxylase activity were also observed but were slightly less than the increases in immunoreactive P-450(17 alpha), suggesting that not all of the protein was enzymatically active. Comparatively, immunoreactive P-450(21) was increased only 1.3-fold. Antibodies to porcine P-450(17 alpha) and bovine P-450(21) reacted monospecifically with the homologous rabbit and guinea pig proteins as judged by the detection of single bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Inhibition studies showed that in an assay using 125 micrograms per ml of microsomal protein ACTH-stimulated rabbit adrenal 17 alpha-hydroxylase activity was inhibited 72% at a 100 mg per ml concentration of the anti-porcine P-450(17 alpha); however, 47% inhibition was observed at the same concentration of anti-bovine P-450(21). Pre-immune IgG had no effect. Molecular weight, Mr, determinations by SDS-PAGE showed both rabbit and guinea pig P-450(17 alpha) to be 52 kDa; rabbit P-450(21), 54 kDa; and guinea pig P-450(21), 49 kDa.(ABSTRACT TRUNCATED AT 250 WORDS)
 ...
PMID:ACTH-induced increases in rabbit adrenal immunoreactive P-450(17 alpha) and P-450(21). 215 89

Using extracts of AtT-20 cell nuclei, protein binding sites on the POMC gene 5'-flanking region were examined with an exonuclease protection approach. One such binding site, located from -119 to -106 bp upstream from the mouse POMC gene transcription initiation site, which exhibited a close homology to the activator protein-2 (AP-2) site [1]. A double-stranded oligonucleotide containing this site was subsequently used in gel shift assays to demonstrate AP-2 consensus sequence binding activity in extracts of AtT-20 cell nuclei. Gel shift competition experiments using both homologous and heterologous competitor DNA sequences revealed that the AP-2 like factor(s) exhibited specific binding to the mouse AP-2 consensus sequence. Furthermore, AP-2 factor binding was also modulated by a CTF/NF1-like factor. Pretreatment of AtT-20 cell nuclear extracts with alkaline phosphatase prior to inclusion in gel shift assays led to a reduction in the intensities of AP-2 factor-specific bands, indicating a potential involvement of protein phosphorylation in AP-2 factor binding in AtT-20 cells.
 ...
PMID:A putative AP-2 binding site in the 5' flanking region of the mouse POMC gene. 233 36

The course of serum zinc (S-Zn), plasma albumin (P-Alb), urinary zinc, serum alkaline phosphatase, and plasma alpha 2-macroglobulin levels was monitored in 14 adult hospitalized patients receiving oral glucocorticoid therapy, about 40-50 mg prednisone daily for various skin diseases. Within 3 days S-Zn decreased slightly from 12.6 +/- 2.3 mumol/liter (mean +/- SEM) to 11.1 +/- 2.5 mumol/liter. Then the level rose to about 14-15 mumol/liter and remained elevated, but within the normal range for the next 2 weeks. The P-Alb level showed parallel fluctuations although less pronounced. The S-Zn/P-Alb ratio increased from 0.024-0.029. No consistent patterns could be seen in the fluctuations occurring in the additional parameters studied. The possible role of ACTH on the S-Zn regulation is discussed.
 ...
PMID:Serum zinc levels during oral glucocorticoid therapy. 242 19

Mouse monoclonal antibodies (MAbs) have been produced against porcine ACTH and tested for their immunocytochemical utility. Ten out of 12 MAbs reacted with formaldehyde-fixed human ACTH[1-39] and fragments thereof. Cytochemical fragment testing revealed that 6 of the 10 MAbs recognized epitopes in the vicinity of the region where porcine ACTH differs from mouse ACTH (amino acids 26, 29 and 31). Both tissue and cytochemical model data indicate that many of the MAbs detected porcine ACTH with somewhat higher potency than human and rat ACTH (rat ACTH[1-39] is identical to mouse ACTH[1-39]). MAbs Nos. 5, 8 and 12, in particular, revealed a highly satisfactory signal to noise ratio also in formalin-fixed, paraffin-embedded specimens. Most of the MAbs were potent in detecting both the high concentrations of ACTH congeners in corticotrophs and melanotrophs and the lower concentrations of such peptides in human antropyloric gastrin cells. Blocking of tissue endogenous peroxidase activity reduced reactivity towards the MAbs. This could be circumvented by use of biotinylated primary antibodies combined with avidin/streptavidin-alkaline phosphatase detection. Availability of MAbs and of the corresponding synthetic antigen also made some quantitative comparisons and analyses of appropriate control procedures possible.
 ...
PMID:Immunocytochemical characterization of mouse monoclonal ACTH antibodies with a note on staining conditions and control procedures. 247 67

The presence of bovine pituitary intermediate lobe peptides in intraglandular colloid, the holocrine secretion of intermediate lobe cells, is explored by ELISA. Intraglandular colloid collected immediately after sacrificing the animal, is placed in phosphate buffered saline, pH 7.6. This material is homogenized, centrifuged to remove extraneous tissue, lyophilized and stored at -20 degrees C. ACTH in intraglandular colloid is measured by competitive ELISA. Human ACTH (1-24) is used in the preparation of the solid phase antigen and as the standard for competition. The antibody is rabbit anti-human ACTH (1-24), and the alkaline phosphatase conjugate is goat anti-rabbit IgG with p-nitrophenyl phosphate as substrate. It is concluded that ACTH is present in bovine pituitary intraglandular colloid of intermediate lobe origin and that the colloid may serve as a transport medium for intermediate lobe materials.
 ...
PMID:ACTH in bovine pituitary intraglandular colloid, the holocrine secretion of intermediate lobe cells. 283 85


<< Previous 1 2 3 4 5 6 7 Next >>