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)

Lymphocytes in the stroma and lymphatics of the extra-cortical central area (ECCA) of the guinea pig thymus have been studied with light microscopy, quantitative microscopy, colchicin-induced mitotic arrest, EA (IgG) and EA (IgM) C adherence, surface immunoglobulins (Ig total, IgM, IgG), alkaline phosphatase activity and the effect of cyclophosphamide administration. The results suggest, that AP-positive, SIg-positive EAC-negative lymphocytes in the ECCA proliferate and maturate into AP-negative, SIg-positive, EAC-positive lymphocytes. The latter leave the thymus through the lymphatics. Calculations show that daily 8-9 X 10(6) B lymphocytes are produced in the ECCA and leave this area through the lymphatics. As this number is more than 10% of the number of T cells which daily leave the thymus, we conclude that in the ECCA a considerable number of B lymphocytes is produced.
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PMID:Production and traffic of B lymphocytes in the extracortical central area of the guinea pig thymus. 34 32

Point mutation in the nucleotide sequence of the structural genes for the TEM-type penicillinases can broaden their substrate spectrum towards all beta-lactams except cephamicins and imipenem. We describe here hybridization techniques for the detection of point mutations by non-radioactive oligonucleotide probes with plasmid DNA carrying bla T genes immobilized in polystyrene microwells. After hybridization in discriminating conditions with corresponding biotinylated oligonucleotide probes, the hybrids were detected by using a streptavidin-alkaline phosphatase conjugate and a fluorogenic substrate, 4-methylumbelliferyl-phosphate. The adsorption of DNA to microwells used in the present work was found to be independent of Mg2+ and Na+ concentrations. By this method, less than 3 fmols of target DNA were sufficient for the detection of point mutation.
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PMID:Detection of point mutation in bla T genes of Enterobacteriaceae by biotinylated oligonucleotide probes using microwell hybridization and enzymofluorometric method. 154 33

The cytocompatibility of a polyepoxy resin (Elf Aquitaine) has been studied using both cell lines and human differentiated cell cultures. The human models were gingival fibroblasts and bone osteoblasts, while the cell lines were Hela cells and 3T3 Balb/c cells. Basal cytocompatibility was assessed by estimation of the cell proliferation, total cell protein content, cell membrane sub-lysis, and cell attachment and spreading. Specific cytocompatibility concerning human osteoblasts, from both alveolar and trabecular bone, was determined by measuring the intracellular alkaline phosphatase activity. Resin colonization by the cells was studied by both TEM and SEM. The behaviour of the two cell lines reveals a significant level of discrepancy, whereas the behaviour of human cells, whatever the model, is comparable; however, osteoblasts look more sensitive. Moreover, the results show that this epoxy resin exhibits a moderate cytocompatibility which could be the result of the cytotoxicity of early released products, associated with the considerable surface roughness.
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PMID:In vitro evaluation of an epoxy resin's cytocompatibility using cell lines and human differentiated cells. 186 79

Biotination of proteins is a post-translational modification that requires a folded acceptor domain. We previously showed that an acceptor domain fused to the carboxyl terminus of several cytosolic proteins results in biotinated fusion proteins in vivo. We now show that proteins encoded by translational gene fusions of two periplasmic proteins, alkaline phosphatase and TEM beta-lactamase, to carboxyl-terminal biotin-accepting sequences are biotinated and exported by Escherichia coli. Expression of the alkaline phosphatase fusion protein in wild type strains resulted in inefficient biotination of the fusion product. This result was due to the rapid export of the acceptor protein before biotination could occur since a very large increase in biotinated fusion protein levels was observed in strains lacking the SecB chaperone protein. The beta-lactamase fusion protein was biotinated but was only stable in strains lacking the DegP periplasmic protease. Both biotinated fusion proteins accumulated in the culture medium in strains possessing defective outer membranes. These results indicate that the export machinery can accommodate both a post-translational modification and a protein domain previously folded into its mature conformation in vivo.
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PMID:Escherichia coli exports previously folded and biotinated protein domains. 205 Jun 59

The enzyme TEM beta-lactamase constitutes a versatile gene-fusion marker for studies on membrane proteins and protein export in bacteria. The mature form of this normally periplasmic enzyme displays readily detectable and distinctly different phenotypes when localized to the bacterial cytoplasm versus the periplasm, and thus provides a useful alternative to alkaline phosphatase for probing the topology of cytoplasmic membrane proteins. Cells producing translocated forms of beta-lactamase can be directly selected as ampicillin-resistant colonies, and consequently a beta-lactamase fusion approach can be used for positive selection for export signals, and for rapid assessment of whether any protein expressed in Escherichia coli inserts into the bacterial cytoplasmic membrane. The level of ampicillin resistance conferred on a cell by an extracytoplasmic beta-lactamase derivative depends on its level of expression, and therefore a beta-lactamase fusion approach can be used to directly select for increased yields of any periplasmic or membrane-bound gene products expressed in E. coli.
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PMID:Beta-lactamase as a probe of membrane protein assembly and protein export. 207 55

Human deciduous teeth undergoing physiologic root resorption were extracted and fixed with a mixture of formaldehyde and glutaraldehyde and processed for scanning (SEM) and transmission (TEM) electron microscopy, and for acid (ACPase) and alkaline phosphatase (ALPase) cytochemistry. The resorbant organ, rich in odontoclasts, cementoblasts, fibroblasts, and macrophages, formed prominent resorption lacunae in root dentin. SEM observations of resorption lacunae treated with trypsin solution showed islands of newly-formed cementum matrix in part of the resorbing dentin surfaces. Such cementum consisted of bundles of densely-arranged collagen fibrils and, in part, contained forming cementocytic lacunae and canaliculi. Active cementoblasts adjacent to odontoclasts on resorbing dentin surfaces showed cuboidal outlines and were characterized by the presence of numerous cisterns of rough endoplasmic reticulum, well-developed Golgi complexes, secretion granules, and many mitochondria. They sometimes formed a thin layer of cementoid and/or cementum matrix upon the resorbing dentin surface. These cementoblasts had ACPase-positive lysosomes in the cell bodies and exhibited intense ALPase activity along the plasma membranes of whole cell surfaces. These results suggest that, during root resorption, 1) active cementoblasts are present adjacent to active odontoclasts and 2) these cementoblasts are involved in remodeling the resorbing dentin surfaces.
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PMID:Possible role of cementoblasts in the resorbant organ of human deciduous teeth during root resorption. 214 74

For the improvement of the adult osteoblast culture, the osteoblasts of young adult rabbit endosteal from long bones were isolated by collagenase digesting separation. 0.1% of type-I collagen precoated culture flasks were used as substrate for isolated bone cell growth. Morphological examination of cultured cells under a phase-contrast microscope, SEM and TEM observations showed a structure similar to osteoblast in vivo. Histochemical examination of alkaline phosphatase demonstrated 97% purity of cultured osteoblasts. The presence of calcium deposit activity in cultured cells was demonstrated by Van Kossa stain. High activity of alkaline phosphatase and inorganic pyrophosphatase in cultured osteoblasts as determined by biochemical analysis. High calcium uptake in cultured osteoblasts was demonstrated by radioisotope labelled 45CaCl12. According to these methods, it was indicated that the cells isolated from young rabbit long bone endosteal were osteoblast-like and still maintained their biological function. Our system for culturing osteoblast-like cells is a successful attempt in growing bone tissue in vitro starting from isolated bone cells. Therefore, this modified method for bone cell culture on collagen precoated culture flasks could be used as the experimental model in studies concerning the osteoblasts in vitro.
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PMID:Functional evaluation of cultured rabbit osteoblast-like cells. 255 21

The cytochemical localization of enzymatic activity by means of backscattered electron imaging (BEI) is reviewed and the application of BEI to changes in acid phosphatase and ATPase distribution during physiological (programmed) cell death in Heliothis midgut is explored. Programmed cell death entails the release of nascent free acid phosphatase as extracisternal hydrolase. This shift can readily be detected by means of the atomic number contrast imparted by BEI of the lead phosphatase reaction product, thus enabling the distribution of dying cells to be mapped. BEI is particularly useful in this context as it allows the examination of bulk specimens at low magnification. Death of cells is also accompanied by a collapse in ATPase activity which shows up as cytochemically negative areas in the X-ray microscope and by means of BEI. Acid phosphatase in normal cells is localized in the apical microvilli and lysosomes. Senescent or dying cells, however, clearly show a basally situated free hydrolase which migrates throughout the cell. Parallel TEM results confirm that this enzyme is ribosomal and extracisternal rather than lysosomal in origin. ATPase activity is largely limited to the apical microvilli, although there is some activity associated with the basal plasma membranes. The apical ATPase, however is partially resistant to ouabain. Young and mature cells are positive although in the latter case some microvilli may be lost as the cells acquire a negative cap or dome. Inhibition by bromotetramizole indicates that apical activity is not to any significant extent contributed to by alkaline phosphatase. Degenerate or dead cells are negative and can be seen as a mozaic of "black patches" among normal cells when imaged by means of BEI or X-ray microscopy.
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PMID:The use of backscattered electron imaging, X-ray microanalysis and X-ray microscopy in demonstrating physiological cell death. 297 76

The histochemistry and ultrastructure (SEM and TEM) of the spermatheca of Biomphalaria glabrata was investigated to elucidate the function of this organ and to compare its structure and function to similar organs found in other species. The spermatheca has a debris-filled lumen surrounded by a thin wall of tissue. The cells adjacent to the lumen are of three columnar epithelial cell types. Two cell types have abundant microvilli and mammalian cell-like organelle distribution and morphology. The above cell types differ in the electron density of their cytoplasms, nuclear morphologies, and organelle content. The third cell type differs from the other two in its cytoplasmic makeup. However, the most distinctive difference is the presence of large numbers of cilia at the apical surface with no evidence of microvilli. These columnar cells rest on a basal lamina adjacent to a two to three cell thick muscle layer. The entire organ is surrounded by an adventitia of unusual morphology. Histochemical investigation demonstrated that DNAase, RNAase, and protease are present in the lumen, alkaline phosphatase is associated primarily with the microvilli, small amounts of acid phosphatase are concentrated in the midcell area of the columnar epithelium, and ATPase activity is localized in the muscle cells and just below the absorptive surface of the microvillous cells. The luminal contents and adventitial areas are Sudan Black B positive, all areas of the lumen and organ wall are PAS positive, the cell nuclei and amorphous masses in the lumen showed Feulgen staining, and large vesicles in the columnar cells were Oil Red O positive. Apparently, the spermatheca of B. glabrata is both a digestive and absorptive structure. Although this organ shares functional similarities with those found in opisthobranchs and terrestrial pulmonates, the epithelia of the spermatheca differ dramatically in these groups.
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PMID:Structure and function of the spermatheca in a snail host of schistosomiasis, Biomphalaria glabrata. 364 39

In this study, Sprague-Dawley rats were exposed in a TEM chamber to 20-MHz (HF-band) continuous-wave radiofrequency radiation (RFR) for 6 hr/day, 5 days/week up to 6 weeks. The average E-field intensity was 2686 +/- 164 V/m (mean +/- SD) and the calculated specific absorption rate was 0.3 W/kg. Randomly sampled rats killed on Days 8, 22, 39, and 42 after initiation of exposure showed no statistically significant differences from controls for body mass, spleen cell density, erythrocyte and leukocyte counts, hematocrit, hemoglobin, methemoglobin, erythrocyte fragility, bilirubin, creatinine, SGPT, alkaline phosphatase, calcium, sodium, potassium, and spleen cell chemiluminescence. Splenic mass differences were statistically significant (p less than 0.05) only on Day 22. Spleen to body mass ratios differed significantly between exposed and control groups on Days 22 and 39 (P less than 0.05 and P less than 0.025, respectively). Histologic examination of the rats revealed the successive accumulation of phagocytic cells, lymphoid proliferation, development of lesions, and tissue necrosis characteristic of respiratory mycoplasmosis. In a followup experiment, a separate set of rats was exposed for 6 weeks to identical levels of RFR. No significant differences were found in splenic parameters and spleen cell peroxidative activity. Histologic examination of these animals revealed no evidence of mycoplasma infection. The observed differences between exposed and control animals of the first experiment appear to have resulted from subclinical respiratory mycoplasmosis rather than exposure to RFR.
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PMID:Effects of 20-MHz radiofrequency radiation on rat hematology, splenic function, and serum chemistry. 402 74


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