Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

A split-flow, air-lift bioreactor for the cultivation of insect cells to produce recombinant protein is described. It can be used advantageously with attached cell systems. This bioreactor incorporates two sections: a rise and a downcomer. Trichoplusia ni BTI-Tn 5B1-4 cells are grown on a support material of glass beads or microcarriers placed in the downcomer. This cell line is more productive than other commonly used insect cell lines, but it has the disadvantage of being difficult to use at large volumes since it is not easily adaptable to suspension culture. Adequate oxygen demand is supplied by sparging without direct exposure of cells to air bubbles. Nutrients are supplied convectively to the attached cells on support material as the fluid flows through the downcomer. The split-flow, air-lift bioreactor appears to be suitable for insect cell culture and is potentially scalable. It can provide a high surface-to-volume ratio and can be operated in batch or continuous mode. A lab-scale prototype bioreactor has been constructed and tested for the production of a secreted, glycosylated recombinant protein (human alkaline phosphatase) or seAP using an Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) vector. With a ratio of riser cross-sectional area to downcomer cross-sectional area of 1, an aspect ratio of 4.4, an air-flow rate of 54 mL/min in the riser, and a bed of 2400 3-min nonporous glass beads, 10.7 micrograms/mL of seAP was produced using an MOI (multiplicity of infection or the ratio of plaque-forming units to cells) of 10.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Production of human alkaline phosphatase, a secreted, glycosylated protein, from a baculovirus expression system and the attachment-dependent cell line Trichoplusia ni BTI-Tn 5B1-4 using a split-flow, air-lift bioreactor. 776 58

A method is introduced for hybridizing large numbers of DNA samples against large numbers of DNA probes on a single support membrane. Denatured DNA from up to 43 samples was fixed in separate lanes on a single membrane mounted in a Miniblotter 45. The membrane was then rotated 90 degrees in the same device, which enabled simultaneous hybridization with 43 different DNA probes. Hybridizations were also performed on lysates of bacterial cells blotted to membranes. A MiniSlot device allowed lysates loaded in parallel channels to be aspirated through the membrane, depositing horizontal lanes on the membrane surface. Hybridizations were performed in vertical lanes with either digoxigenin-labeled whole genomic probes or 16S rRNA-based oligonucleotide probes directly conjugated to alkaline phosphatase. The method permits the simultaneous determination of the presence of multiple bacterial species in single or multiple dental plaque samples, thus suggesting its usefulness for a range of clinical or environmental samples.
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PMID:"Checkerboard" DNA-DNA hybridization. 783 43

The effect of oral epidermal growth factor (EGF) on histological and biochemical changes in epithelium in the small intestine was studied in colostrum-deprived neonatal pigs. Forty-eight pigs were infected at 4 days of age with 2 x 10(7) plaque-forming units of porcine group A rotavirus and orally fed a simulated sow-milk diet supplemented with 0.0, 0.5, or 1.0 mg/L recombinant human EGF. Sixteen noninfected pigs were fed a diet without EGF supplementation. Infected pigs developed severe diarrhea; they also consumed 25% less food and gained 60% less weight than noninfected pigs. Pigs were killed 8 days postinfection to collect samples at seven equidistant points in the small intestine. Rotavirus infection decreased villus height by 37% and reduced specific activity of lactase by 54%, of leucine aminopeptidase by 43%, and of alkaline phosphatase by 54% in the small intestine, compared with noninfected pigs. Only the supraphysiological dose of EGF (1.0 mg/L) consistently increased villus height in the proximal and mid-small intestine and lactase-specific activity in the mid-small intestine of rotavirus-infected pigs. However, this dose was only partially effective in restoring intestinal mucosal dimensions and enzyme activities. Supplemental EGF did not hasten the resolution of diarrhea. These data indicate that high physiological levels of EGF are beneficial in stimulating recovery of epithelium in the small intestine following rotavirus infection.
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PMID:Effect of orally administered epidermal growth factor on intestinal recovery of neonatal pigs infected with rotavirus. 787 90

Interbacterial binding is considered an important colonization mechanism for many of the organisms that inhabit dental plaque. Porphyromonas gingivalis, a periodontal pathogen, can adhere to species that comprise early plaque, such as Streptococcus gordonii. In this study, the molecules of S. gordonii G9B that mediate binding to P. gingivalis were investigated. Biotinylated surface molecules of S. gordonii were extracted and mixed with P. gingivalis cells. Interactive streptococcal components were identified by SDS-PAGE of the P. gingivalis cells followed by electroblotting, and visualization of the adsorbed streptococcal molecules with streptavidin-alkaline phosphatase. S. gordonii molecules of 45 kDa and a doublet of 62/60 kDa were observed to bind to P. gingivalis. Polyclonal antibodies raised to the 62/60 kDa proteins inhibited the binding interaction. These antibodies demonstrated an antigenic relationship between the 62/60 kDa molecules and the 45 kDa protein. Both molecules were also antigenically related to, and may be breakdown products of, a larger molecule of 170 kDa which is antigenically related to the P1 antigen of S. mutans. Cloning and expression in Enterococcus faecalis of the gene for the P1-like molecule from S. gordonii M5 resulted in a phenotype that expressed the 62/60 kDa and 45 kDa antigens and was capable of binding to P. gingivalis. These results suggest that a P1-like molecule in S. gordonii is involved in adherence to P. gingivalis. Processing of the P1-like molecule into smaller fragments of 62/60 kDa and 45 kDa may be required for binding activity.
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PMID:Molecules of Streptococcus gordonii that bind to Porphyromonas gingivalis. 801 3

Using a recently developed chemiluminescent assay enabling alkaline phosphatase (ALP) quantification in nanolitre volumes of gingival crevicular fluid (GCF), we have investigated GCF ALP levels in health and in the presence of gingivitis. In gingival health, there was a site-specific pattern of ALP concentration with higher enzyme concentrations around the upper and lower anterior teeth. Furthermore, clinically normal sites that had been subjected to different levels of plaque control produced significantly different ALP levels, (p < 0.03). This indicates that biochemical components of GCF may be used to measure subclinical inflammatory status. The ratio of GCF to serum ALP varied from 6:1 to 11:1, suggesting that a major source of the enzyme is through local production. The main cross-sectional study of 30 patients with gingivitis (276 sites) demonstrated that total GCF ALP levels, collected over a 30-s sampling time were higher for a gingival index of 1 than of 0 (p < 0.014). There was no significant relationship between total GCF ALP and plaque levels of the enzyme, and analysis of plaque within the study group demonstrated very low levels of ALP, indicating that the enzyme is likely to be largely derived from the periodontal tissues. The ratio of GCF ALP levels to those of saliva within individuals was 530:1, thereby eliminating saliva contamination as a risk, when total GCF ALP is being measured.
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PMID:Site-specific alkaline phosphatase levels in gingival crevicular fluid in health and gingivitis: cross-sectional studies. 808 43

Previous studies in our laboratory demonstrated messenger RNA for bone morphogenetic protein-2a in human calcified plaque, suggesting that arterial calcification is a regulated process, similar to osteogenesis. To further test this hypothesis, we have isolated and cloned a subpopulation of cells from bovine aortic media that show osteoblastic potential. These novel cells are primarily distinguished from smooth muscle cells by expression of a surface marker preliminarily identified as a modified form of the ganglioside sialyl-lactosylceramide (GM3). Osteoblastic potential was indicated by high levels of alkaline phosphatase and collagen I, expression of osteopontin and osteonectin (SPARC), and production of bone-specific osteocalcin and hydroxyapatite. Cultures of these cells were stimulated to form increased numbers of calcium-mineral-producing nodules by the oxysterol 25-hydroxycholesterol as well as by transforming growth factor-beta 1, both known to be present in atherosclerotic lesions. The stimulation of calcifying vascular cells in the artery wall by these two factors suggests a possible mechanism for the colocalization of calcification with atherosclerosis in vivo.
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PMID:TGF-beta 1 and 25-hydroxycholesterol stimulate osteoblast-like vascular cells to calcify. 818 41

Terephthalic acid at 20 mg/kg/day in lowered serum cholesterol and triglyceride levels in rats. The cholesterol content was lowered in the lipoprotein fractions. The effects of the agents on de novo lipid synthesis showed that similar enzymes were affected in rat liver and small intestinal mucosa cells as when compared to in vitro tissue culture cells from rats and humans, e.g. reduction of acyl CoA cholesterol acyl transferase and elevation of neutral cholesterol ester hydrolase activities suggest that net cholesterol esters deposition in foam cells should be reduced and plaque growth should be slowed. The suppression of LDL receptor binding and degradation by the drug suggest that less apoB lipoproteins are taken up by peripheral tissues. The elevated HDL receptor binding and internalization in the liver suggest that the drug accelerates cholesterol return to the liver. Additional studies show that cholesterol and bile acid secretion in the bile is elevated. However, the bile acids secreted are not lithogenic. Acute toxicity studies show that the agent appears to be safe in rodents. Two observations of increased serum alkaline phosphatase levels and increased liver vacuolation suggest some alteration of hepatic cell morphology, which requires further investigation.
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PMID:Terephthalic acid in Sprague-Dawley rats as a hypolipidemic agent. 844 23

We developed a colorimetric microtiter plate-based assay for the detection and quantification of polymerase chain reaction-amplified DNA fragment specific for Actinobacillus actinomycetemcomitans. We amplified the 396-bp leukotoxin-specific DNA fragment by using two oligonucleotide primers, one carrying a biotin group at the 5' end and another one with a digoxigenin at the 5' end. Following amplification, the biotinylated polymerase chain reaction products were applied to a microtiter well precoated with avidin. The colorimetric detection and quantification were achieved by an enzyme-linked immunosorbent assay using alkaline phosphatase-conjugated anti-digoxigenin antibody. The detection limit of the colorimetric assay was found to be as little as 500 fg of purified A. actinomycetemcomitans DNA and as few as 50 A. actinomycetemcomitans. Therefore, this colorimetric assay was able to estimate the amount of A. actinomycetemcomitans in subgingival plaque samples. We concluded that the colorimetric assay of the PCR product is a very useful method not only to detect the presence of A. actinomycetemcomitans but also to quantify the amount of A. actinomycetemcomitans in large numbers of subgingival plaque samples.
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PMID:Colorimetric microtiter plate based assay for detection and quantification of amplified Actinobacillus actinomycetemcomitans DNA. 860 46

This study examines acid and alkaline phosphatase activities in gingival crevicular fluid (GCF) to learn whether bone turnover dynamics can be monitored in human subjects during orthodontic tooth movement. Three female subjects were observed longitudinally to assess tooth movement, plaque, and inflammation. For each subject, one randomly selected premolar served as the control and was not treated, and another was moved buccally with 100 gm of force. The GCF was collected weekly and assayed for phosphatases. Alkaline phosphatase peaked between the first and third weeks, followed by an increase in acid phosphatase between the third and sixth weeks. After the first week, tooth movement averaged 0.9 mm. Additional 0.9 mm of movement occurred during the next 3 weeks, followed by 1.4 mm during weeks 4 to 6. Thirty additional patients, randomly divided into headgear/biteplate, bionator, and control groups, were also sampled cross-sectionally at the maxillary first molars. The GCF phosphatase activities were assessed as functions of location on the tooth, treatment modality, duration of treatment, gingival inflammation, and plaque accumulation. The plaque index did not show a relationship to either acid or alkaline phosphatase activity on the mesial or distal in the treated groups. However, alkaline phosphatase increased with inflammation on the distal in treated groups and acid phosphatase was consistently higher on the mesial than on the distal in the treatment groups. Alternating peaks of acid and alkaline phosphatase were found in the GCF of treated teeth as functions of treatment duration. The sequence of these changes is similar to that reported for alveolar bone turnover in a rodent orthodontic tooth movement model. We conclude that phosphatase activities in GCF may be a useful means for monitoring tissue responses to orthodontic treatment.
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PMID:The measurement of acid and alkaline phosphatase in gingival crevicular fluid during orthodontic tooth movement. 860 74

In chemostat culture, the microaerophilic, CO2 requiring, gingival-plaque-associated bacterium Capnocytophaga gingivalis responded to the addition of glucose (1-6 g I-1) by doubling its growth rate and increasing its biomass yield fivefold. The data suggest that the glucose is catabolized by a fully aerobic route. Rather than repressing hydrolytic enzymes which might be associated with pathogenic properties, glucose enhanced the specific activity of aminopeptidase, trypsin-like protease, acid and alkaline phosphatase and alpha-glucosidase in comparison with a control culture grown in a tryptone/thiamin medium. Thus, the supply of glucose could be of importance in maximizing the pathogenic potential of this organism.
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PMID:Capnocytophaga gingivalis: effects of glucose concentration on growth and hydrolytic enzyme production. 876 Sep 30


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