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)

The present study has been undertaken to investigate the efficiency of biotinylated synthetic oligonucleotide probes in detection by in situ hybridization of the mRNAs coding for calcitonin (CT) or calcitonin gene-related peptide (CGRP) in human medullary thyroid carcinomas (MTCs). Tissue sections fixed with formaldehyde were hybridized with 45-base long oligonucleotides, specific for CT or CGRP mRNA. Recombinant DNA probe or synthetic oligonucleotides radioactively labelled with 32P or 35S were used as controls to detect by autoradiography the corresponding mRNAs in the tumour cells. Oligonucleotide probes labelled by fixation of one biotin molecule at their 5'-end, or by incorporation of a tail of biotin-11-dUTP at their 3'-end, were used and were revealed by incubation with streptavidin-alkaline phosphatase associated with the corresponding substrate. Each biotinylated probe stained exclusively the cytoplasm of the tumour cells, the CT probe giving a much higher level of staining than the CGRP probe. The same cells were found to contain CT and CGRP mRNAs. Controls performed with either radioactive or biotinylated probes confirmed the specificity of the staining. These results demonstrate that biotinylated synthetic oligonucleotides can be used as efficient tools to investigate gene expression in tissue sections, thus avoiding the various inconveniences connected with the use of radioactive probes, especially bio-hazards, the use of autoradiography, the limited histological resolution, and the delay in obtaining results.
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PMID:Histochemical detection of the messenger RNAs coding for calcitonin and calcitonin gene-related peptide in medullary thyroid carcinomas with radioactive and biotinylated oligonucleotide probes. 233

The activity of acid and alkaline phosphatases was determined in glomeruli and tubules isolated from 14-day old to adult rat renal cortex. Within a nephron fraction, each phosphatase activity produced a specific pattern which was not similar to that in other fractions. In the glomeruli the acid phosphatase activity increased between 14 and 21 days of life and was always higher than in the tubules where it did not significantly change with age. By using tartrate sodium and formaldehyde it was shown that the acid phosphatase isoenzymes are distributed according to a specific pattern which does not change during maturation. The alkaline phosphatase activity in glomeruli and in tubules increases with age but is always lower in the former fraction than in the latter. Furthermore, in young as well as in adult rats, glomeruli were characterized by a higher DNA content and lower RNA and protein/DNA ratios than the corresponding tubular fragments.
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PMID:Activity of phosphatases and cellular contents in glomeruli and tubules isolated from growing rat renal cortex. 241 90

Sections of formaldehyde-fixed paraffin-embedded cortical and hippocampal brain tissue from five cases with senile dementia of Alzheimer type (SDAT) and five cases with Pick's disease (PD) were immunostained with the monoclonal antibodies (mabs) 147, RT 97, BF 10 and 8D8 with and without pretreatment with alkaline phosphatase (AP) or trypsin (Tr). The mabs 147, RT 97 and BF 10 had previously been demonstrated to bind exclusively to phosphorylated epitopes of neurofilament proteins, while mab 8D8 is shown in this report to bind mainly, but not exclusively, to phosphorylated neurofilament epitopes. The mabs RT 97, BF 10 and 8D8, but not 147 stain most, if not all, Pick bodies (PB) and Alzheimer neurofibrillary tangles (NFT). When sections are pretreated with AP or Tr the immunostaining with mab BF 10 is very resistent in both PB and NFT. This resistance of PB and NFT is in contrast to the reduced staining of axons and of swollen cells in PD by the same enzymatic pretreatment. Immunostaining with mab RT 97 of PB and NFT is reduced moderately by AP and considerably by Tr. Only when stained with mab 8D8 is there a discrepancy between PB and NFT in their reaction to the pretreatment with AP: NFT staining with mab 8D8 is not affected, while that of PB is abolished. Thus, in spite of their different ultrastructure, PB and NFT are very similar immunocytochemically and in the accessibility of their phosphorylated epitopes to enzymatic treatment.
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PMID:Alzheimer dementia and Pick's disease: neurofibrillary tangles and Pick bodies are associated with identical phosphorylated neurofilament epitopes. 244 59

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.
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PMID:Immunocytochemical characterization of mouse monoclonal ACTH antibodies with a note on staining conditions and control procedures. 247 67

The purpose of the study was assessment of the effect of an environment contaminated with heavy metals on the activity of certain enzymes of mixed saliva. The activity was determined of total acid phosphatase and phosphatase resistant to tartrate and formaldehyde, alkaline phosphatase, alanine and aspartate aminotransferase, and alpha-amylase. The studied material comprised 110 saliva samples obtained from three groups of children aged 8 years. Group I of 21 children lived in Szopienice, group II of 30 children lived in Miasteczko Slaskie. In both these localities the children were exposed to mean daily concentrations, above the permitted ones, mainly of lead compounds, in lower degree to cadmium and zinc compounds. Environment contamination in Szopienice was greater than in Miasteczko Slaskie. Group III of 59 children living in Lubowice served as controls. In that town the permissible concentrations of these compounds were not exceeded. Statistical analysis of these results showed that the activity of total acid phosphatase in groups I and II, that is in the contaminated areas, was highly significantly greater than in the control group. The activity of alkaline phosphatase was raised only in the saliva in group I. No differences were found in the activity of alpha-amylase and aminotransferases.
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PMID:[Activity of certain salivary enzymes in school children exposed to excessive concentrations of lead and cadmium]. 264 Jun 43

The enamel organ of the growing rat incisor was fixed with a mixture of formaldehyde and glutaraldehyde and processed for ultracytochemical demonstration of Ca- and Mg-activated membrane ATPase by a one-step lead technique at alkaline pH. To inhibit nonspecific alkaline phosphatase, 5 mM levamisole was added to the incubation media. Intense Ca- and Mg-ATPase activity was demonstrated in the cell surfaces of the secretory ameloblasts, except at the proximal and distal junctional complexes and the gap junctions in the lateral and basal cell surfaces. Deep plasma membrane invaginations at the proximal and distal parts of Tomes processes facing interrod- and rod-enamel growth regions exhibited the strongest enzymatic reaction. Mg-ATPase activity was also shown to be present in the plasma membranes of secretory ameloblasts but it was less intense than Ca-ATPase. Except for a slight reaction in the Golgi membranes, all other cell organelles of the secretory ameloblasts and the adjacent enamel matrix were free of enzymatic reaction. However, when the tissues were incubated in media lacking levamisole, a prominent enzymatic reaction was observed in the newly secreted enamel matrix of the rod and interrod growth regions as well as on the plasma membranes of the cells. In maturation ameloblasts of both ruffle-ended and smooth-ended types, a weak reaction for Ca- and Mg-ATPase was restricted to basal cell surfaces facing the papillary cell layer. In tissues incubated in media lacking levamisole, a variable deposition of reaction products was observed in the Golgi membranes, mitochondrial membranes, tubular elements of smooth endoplasmic reticulum in the ruffled border zone, and along the plasma membranes of the ruffled border. Throughout the secretory and maturation stages, a moderate and/or weak enzymatic reaction for both Ca- and Mg-ATPase was seen in the plasma membranes of the cells of the stratum intermedium and the papillary layer when incubated in media with levamisole. Omission of substrate ATP and/or the enzyme activator CaCl2 from the incubation media for Ca-ATPase produced a negative reaction in the tissues examined. When the calmodulin blocker trifluoperazine was administered to the rats intravenously, Ca-ATPase activity was almost completely abolished from the plasma membranes of secretory ameloblasts, but not of other cell types.
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PMID:Ultracytochemical demonstration of ATP-dependent calcium pump in ameloblasts of rat incisor enamel organ. 294 78

We achieved histological detection of the messenger RNAs coding for vasopressin, calcitonin, or calcitonin gene-related peptide by using biotinylated synthetic oligonucleotides, and defined the technical parameters enabling optimal detection of these mRNAs. Oligonucleotides labeled by fixation of one biotin at their 5' end or by addition of a biotin-11-dUTP tail at their 3' end can be used to detect mRNAs, although the latter are more sensitive. Streptavidin-alkaline phosphatase revealed with nitroblue tetrazolium-bromo-chloro-indolyl phosphate as substrate makes possible detection of the biotinylated oligonucleotides. Increasing formaldehyde concentration in the fixative decreases the signal intensity; 1% formaldehyde fixation provides the most intense signal. Several controls, including those with addition of unlabeled oligonucleotides to the hybridization buffer, confirm the specificity of mRNA detection. The sensitivity of the biotinylated probes is identical or lower as compared to the corresponding radiolabeled oligonucleotides. Histological and subcellular resolution is greatly enhanced with biotinylated probes. The rat vasopressin probes stain magnocellular neurons in the supraoptic and paraventricular nuclei and, under optimal conditions, parvocellular neurons in the suprachiasmatic nucleus. Vasopressin mRNA is present in the cytoplasm of the cell bodies and in the roots of certain processes. Calcitonin and calcitonin gene-related peptide mRNA are found co-localized in the cytoplasm of the same tumor cells in human medullary thyroid carcinoma.
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PMID:Histological detection of messenger RNAs with biotinylated synthetic oligonucleotide probes. 325 49

Colorectal epithelium is composed of a variety of cell types, including absorptive, mucous and endocrine cells. All of these cell types are thought to arise from stem cells located at the base of the crypt. However, the factors which control these differentiation pathways are poorly understood. In attempts to establish differentiated in vitro systems, one approach has been to grow primary human colorectal adenocarcinomas as cell lines. Some of these cell lines retain a sufficient number of the differentiated features of their tissue of origin to make them useful experimental systems for studying differentiation. This study describes the characterisation of such a cell line, the HRA-19 line. HRA-19 cells were derived from a primary human rectal adenocarcinoma. The cells grew as monolayers in vitro on tissue-culture plastic and remained pleomorphic even after 150 passages in vitro. Some colonies of cells expressed alkaline phosphatase activity, an enzyme normally expressed in vivo by absorptive cells of the upper crypt and surface epithelium. No evidence of differentiation into goblet or endocrine cells was obtained in monolayer cultures of HRA-19 cells. Xenografts of this cell line contained cells with the ultrastructural characteristics of absorptive and endocrine cells. These endocrine cells exhibited Grimelius silver staining, displayed formaldehyde-induced fluorescence and contained many basally located, electron-dense granules. When grown as monolayers, clones of this cell line retained the heterogeneity with respect to morphology and alkaline phosphatase expression of the parent cell line. It is proposed that this cell line is derived from malignant progenitor cells which retain the ability to differentiate.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Endocrine differentiation by a human rectal adenocarcinoma cell line (HRA-19). 356 97

Male and female albino Wistar rats were exposed to concentrations of 0, 1, 10 or 20 ppm formaldehyde vapour during 6 h/day, 5 days/wk for 13 weeks. Treatment-related changes observed at 20 ppm included in both sexes: stared coats, uncoordinated locomotion and excitation during the first 30 minutes of each exposure, yellowing of the fur, growth retardation, a decreased level of plasma protein, severe and extensive karatinized stratified squamous metaplasia of the nasal respiratory epithelium, and focal degeneration and squamous metaplasia occasionally accompanied by keratinization of the olfactory epithelium; in males only; increased activities of plasma aspartate amino transferase (ASAT), alanine amino transferase (ALAT) and alkaline phosphatase (ALP) and squamous metaplasia of the laryngeal epithelium. Lesions seen at 10 ppm included yellowing of the fur and moderate squamous metaplasia of the nasal respiratory epithelium. The only change observed in three out of twenty 1 ppm exposed animals that might or might not be treatment-related was minimal focal epithelial hyperplasia and squamous metaplasia of the respiratory epithelium lining the nasal septum and maxillary turbinates. No histopathological evidence of hepatotoxicity was detected in any of the formaldehyde-treated groups. An in vivo/in vitro cell proliferation study showed an increase in [3H]-thymidine labeling index of the respiratory epithelium lining the nasoturbinates of rats exposed to 10 or 20 ppm formaldehyde on three successive days, whereas at the 1 ppm level the labeling index was similar to that of controls. It was concluded that under the conditions of the present 13-week inhalation study, formaldehyde at concentrations up to 10 ppm was not hepatotoxic to rats. At the 20 ppm formaldehyde level, a slight effect on the liver of male rats cannot be completely excluded. The study was inconclusive with respect to 1 ppm formaldehyde being a cytotoxic or a no-cytotoxic effect level for the nasal epithelium.
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PMID:Subchronic (13-week) inhalation toxicity study of formaldehyde in rats. 361 96

Photobiotin was used to prepare biotinylated, nonradioactive nucleic acid probes for the detection of the RNA of barley yellow dwarf virus (BYDV) in plant extracts. A 1.7-kb cDNA of the PAV isolate of BYDV in the plasmid pUC8 vector was biotinylated and then used intact or as sonicated double-stranded DNA fragments. Simple methods were developed for the preparation of partially purified nucleic acid extracts of cereals and their spotting, after formaldehyde treatment, onto nitrocellulose membranes. After hybridization, biotin-labelled DNA bound to BYDV RNA on the nitrocellulose was detected with an avidin-alkaline phosphatase conjugate. BYDV RNA was readily detected with a sensitivity similar to that found with the same probe labelled with 32P by nick translation. Healthy plant extracts gave colourless spots.
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PMID:Nonradioactive, photobiotin-labelled DNA probes for the routine diagnosis of barley yellow dwarf virus. 365


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