Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Surface antigens of adult filarial parasite S. digitata was isolated by employing techniques from manual dissection to treatment with detergents. Among the surface antigen preparations (SAPs), the activities of marker enzymes such as alkaline phosphatase, adenosine triphosphatase and 5' nucleotidase were higher with that isolated by triton X-100 technique (SAP2). On SDS-PAGE, the SAP2 has three major proteins with molecular weights 17, 29 and 36 KD which were consistent with the PBS soluble cuticular proteins (SAP1). Besides these, few other minor protein bands were also observed with the other SAPs. All SAPs were antigenic and showed positive reaction against antiserum to SAP2, and the results confirmed the SAP2 as a better preparation. The release of 29 KD surface protein during in vitro culture of adult parasite and its cross-reactivity with antiserum to surface antigens revealed the possible natural shedding of surface molecules into the host system.
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PMID:Isolation and analysis of surface antigens of filarial nematode Setaria digitata. 176 14

BALB/C female mice were given different dosages of TNF in 0.1 ml sterile PBS containing 1% human serum albumin. Control mice were injected with PBS and human albumin alone. Autopsy examination was carried out and blood biochemistry studied. The results showed that the LD50 was 6 X 10(7) mu/kg. There were serious hyperemia and inflammation of the organs of dead mice, while other smaller dosages of TNF caused acute toxicity of different degrees, except for the 3 X 10(6) mu/kg dosage. Changes of alkaline phosphatase were significant compared with control. Blood sugar increases correlated with the TNF dosage. Changes of GPT and BUN were insignificant. TNF levels in the sera of humans and rabbits were also studied following TNF injection. The serum level of TNF decreased rather quickly in both animals and patients: about 85% of TNF was lost within 5 min after TNF injection, and no TNF could be detected 6 hrs after injection.
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PMID:[Studies on acute toxicity and serum level changes of tumor necrosis factor]. 253 78

A sensitive enzyme-linked immunosorbent assay (ELISA) to measure antibodies against capsular polysaccharide was developed, based on the enhanced binding of polysaccharide to polystyrene microtitration plates. The wells of the microtitration plate were primed with an adipic acid dihydrazide derivative of bovine serum albumin (AH-BSA) (100 micrograms/mL, 0.01 M NaPO4-0.14 M NaCl, pH 7.2 (PBS]. Capsular polysaccharide, the glucuronoxylomannan of Cryptococcus neoformans serotype A, was oxidized with NaIO4 for 5 min; the reaction was then quenched with ethylene glycol. The partially oxidized polysaccharide was dialyzed vs. PBS, and its concentration was adjusted to 50 micrograms/mL with PBS. This solution (100 microL/well) was covalently bound to the AH-BSA primed microtitration plates through formation of a Schiff base between the hydrazide group on the AH-BSA and the aldehyde groups on the polysaccharide. Antimouse IgG-alkaline phosphatase conjugate was used in an indirect ELISA to measure captured murine monoclonal antibodies directed against glucuronoxylomannan. Mean absorbances, after 15 min, were 0.13 in negative control wells, and greater than 0.7 in test wells. No intermediate steps were required to block nonspecific binding of antibody.
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PMID:Enhanced binding of capsular polysaccharides of Cryptococcus neoformans to polystyrene microtitration plates for enzyme-linked immunosorbent assay. 306 47

Immunohistochemical and histochemical methods are increasingly used and their application in surgical pathology is obvious. Especially we used these methods on bone marrow core biopsies. Optimal and comparable results have been obtained by using different methods after halving the biopsy cores longitudinally and/or transversally. The two halves were used for cytologic imprints. Two parts of the biopsy cores were embedded in polymethacrylate at low temperature (-20 degrees C). The methacrylate-embedded biopsy part for routine histology was fixed in Schaffer's solution (methanol-formalin-fixative). The methacrylate-embedded undecalcified section of 4 microns may be stained by most stains commonly employed in routine histopathology after removal of the plastic. The sections are virtually free of artefacts such as shrinkage and swelling in the light microscope. The second methacrylate-embedded part of biopsy cores was fixed in 2% paraformaldehyde with 5% sucrose in 0.02 M phosphate buffer (pH 7.4) and dehydrated in ethyleneglycolmonobutylether. All procedures were carried out at 4 degrees C. This method permits the use of immunohistochemical and histochemical procedures. The immunohistochemistry was carried out at sections of 4 microns after removal of the plastic with methoxide and use of proteolytic enzyme (0.1% alpha-chymotrypsin) to unmask antigens in sections. Surface and intracellular immunoglobulins were very well detected with the indirect FITC method. The histochemical procedures are carried out at sections of 7-8 microns after removal of plastic with xylene and toluol. The sections were incubated for specific esterase and nonspecific esterases, acid and alkaline phosphatase and then examined by light microscopy. A third part of biopsy cores may be immediately frozen, and cryostat sections are stained and evaluated for rapid diagnosis and used for immunohistologic analysis with mono- and polyclonal antibodies (FITC method) and/or histochemical investigations. Imprints of biopsy cores are evaluated for cytological, cytochemical and/or immunocytological analysis with mono- and polyclonal antibodies (FITC method). The cryostat sections and the imprints are fixed for all methods with 2% paraformaldehyde and 5% sucrose in PBS (0.02 M, pH 7.4) at 4 degrees C for 30 minutes. The best diagnostic results were obtained in the myelo- and lymphoproliferative disorders using the combination of methods described here. Examples were demonstrated.
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PMID:[Immuno- and enzymehistochemical studies of methacrylate-embedded biopsy material, especially iliac crest biopsies]. 313 12

Intense alkaline phosphatase (ALPase) activity has been localized in the outer plexiform layer of the developing chick retina. To elucidate the functional significance of this enzymatic activity, we have injected an ALPase inhibitor, levamisole, into embryonic eyes on either the 13th or 15th day of incubation. The retina was fixed between the 15th and 20th day of incubation and examined by electron microscopy. Levamisole injection on the 13th day caused various morphological alterations in retinal development, including the appearance of solitary photoreceptor cells in the subretinal space as well as folding of both the outer plexiform and outer nuclear layers. Pedicles of photoreceptor cells in the outer plexiform layer displayed rather smooth configurations with a reduced number of invaginations by post-synaptic neurites. The outer plexiform layer was thinned and the neuritic extensions in this layer appeared much less developed than in the control (PBS-injected) retina. Photoreceptor outer segments were seldom observed. Besides these alterations, layers of optic fibers and ganglion cells were also affected, as shown by evidence of degeneration in the ganglion cells and thinning of the nerve-fiber layer. Injection of levamisole into day 15 embryonic eyes exerted less influence on retinal development, but some photoreceptor cells were still found in the subretinal space. Some of these observations have been reported in the retinas of aged normal animals or in retinas with hereditary or induced retinal dystrophy. It is suggested that ALPase activity in the outer plexiform layer of the developing chick retina may be important for the onset of normal development of synapses in the outer plexiform layer and differentiation of the photoreceptor cells.
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PMID:Alterations in the differentiation of chick retina caused by an intraocular injection of an alkaline phosphatase inhibitor. 380 58

We describe the development of a simple and highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) for measuring IgG and IgM anticardiolipin antibodies (ACA). Microtitre plates were coated with cardiolipin at a concentration of 45 micrograms/ml by evaporation under nitrogen. Non-specific binding of diluted sera was eliminated by blocking of plates with 10% fetal calf serum in phosphate buffered saline (PBS/FCS) for 2 h. Then sera (100 microliters) at a dilution of 1:100 were incubated in the wells for 1 h. Affinity purified goat anti-human IgG or IgM (100 microliters) at a concentration of 1 microgram/ml was subsequently added and allowed to incubate for 1 h; detection of ACA was achieved using an alkaline phosphatase conjugated rabbit anti-goat IgG reagent by reading the colorimetric yield at 405 nm after incubation with substrate. Reference serum pools were established to study reproducibility of the assay throughout its sensitivity range, and Standard curves were established. The quantitative normal range was 0-9.0 Anticardiolipin ELISA Units (AEU) for IgG and 0-8.0 (AEU) for IgM-ACA. A strong correlation was found between the ELISA and radioimmunoassay methods for measuring ACA of both IgG and IgM classes. Results from 65 patients with systemic lupus erythematosus (SLE) and 45 patients with seropositive rheumatoid arthritis are also reported. The advantages of the ELISA method for quantitative determination of ACA levels, should make it a useful and reliable method for clinical and experimental monitoring of patients with SLE and associated autoimmune disorders.
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PMID:Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results. 408 54

Microvillous membrane fractions from human term placentae were prepared by differential centrifugation. Extration of membranes with PBS-EDTA or KCI removed soluble cytoplasmic components and serum proteins excepting trace amounts of albumin and transferrin. PAGE-SDS revealed 11 components in the Triton solubilized crude fraction after PBS-EDTA extraction. Membrane components solubilized with Triton were not fractionated by gel filtration on Bio-Gel A-50 m but DEAE-cellulose chromatography partially resolved these components. Three fractions were obtained by stepwise elution of absorbed materials using increasing concentrations of NaCl in the equilibrating buffer. These fractions were characterized using SDS-PAGE. The material unabsorbed to the DEAE contained two components of small molecular weight and one of them showed a positive PAS stain. The first eluted protein peak showed nine components, seven of which stained with PAS. The bulk of glycoproteins with molecular weights greater than 130 000 daltons were found in this fraction. The second eluted peak from DEAE was rich in components with molecular weights less than 42 000 daltons. Four components in this fraction were not identified in the other two ion-exchange fractions. Bands representing mobilities of albumin, transferrin and alkaline phosphatase were observed in DEAE-cellulose fractions; however, 12 components of unknown structure were revealed.
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PMID:Characterization of solubilized microvillous membrane proteins and glycoproteins from human placental syncytiotrophoblast. 723 34

RMP-7, a bradykinin agonist, is a synthetic nonapeptide designed to enhance the delivery of therapeutics to the central nervous system. A sensitive, competitive chemiluminescent enzyme-linked immunosorbent assay (ELISA) for quantifying RMP-7 in human blood samples has been developed. Rabbit antibodies against RMP-7 were produced using the conjugate of RMP-7 to keyhole limpet hemocyanin through glutaraldehyde. Biotinylated RMP-7, conjugated via N-hydroxysuccinimide ester, was used as the tracer. A premixed solution of biotinylated alkaline phosphatase and avidin was used to quantify the tracer, with a dioxetane-based compound as the chemiluminescent substrate. The method involves treating blood samples with organic solvents to precipitate proteins, evaporating the supernatants to dryness, reconstituting residues in PBS and assaying the buffer solutions with the ELISA. The assay, using 1.0 ml of whole blood, has precision and accuracy within +/- 20% over the concentration range 25-800 pg ml-1. There are no significant endogenous interferences. The assay has been successfully used to support clinical trials of RMP-7.
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PMID:A competitive chemiluminescent enzyme-linked immunosorbent assay for the determination of RMP-7 in human blood. 888 12

Differential distributions of alkaline phosphatase (AP) and dipeptydylpeptidase IV (DPPIV) were studied in coronary microvascular endothelial cells. Endothelial cells were obtained by the perfusion of coronary vessels with 0.1% trypsin PBS solution and cultured in uncoated culture dishes. Staining of cultured endothelial cells with AP- and DPPIV-sensitive reagents revealed blue or red staining, respectively. Most colonies showed cells of only one color, blue or red, even at the fifth passage. AP-sensitive cells, which were originally elongated, shortened and widened, proliferating to form monolayer colonies of cobble stone-like cells. AP-stainability became weak with repeated passages. DPPIV-sensitive endothelial cells remained elongated even after repeated passages. The cell shape and stainability seemed to be coupled and maintained through the five passages studied.
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PMID:Different enzyme activities in coronary capillary endothelial cells. 926 49

The aim of the study was to evaluate the quantitative and morphological changes in type II alveolar epithelial cells in the course of cyclophosphamide-induced lung damage. The experiments used 40 Wistar rats, of 170 g body weight. The animals were divided into two experimental groups. Group I animals were given cyclophosphamide (Endoxan-ASTA) in a single intraperitoneal dose of 150 mg/1kg b.w./1 ml PBS. Group II (control) received 1 ml PBS. All the animals were sacrificed after 1, 7, 14 and 28 days following intraperitoneal cyclophosphamide or PBS administration. Morphological examinations of pulmonary tissue were based on ultrastructural analysis in the transmission electron microscope. Quantitative studies of type II cells were performed basing on sections stained for alkaline phosphatase. The results of the quantitative analysis showed statistically significant alterations in the number of type II alveolar epithelial cells after 14 and 28 days following cyclophosphamide administration, compared with the respective control groups. The increase in the number of type II cells was accompanied by the intensification of fibroplasia processes within the interstitium of the interalveolar septa of the lungs. Ultrastructural exponents of active participation of type II cells in fibroplasia processes were found.
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PMID:Type II alveolar epithelial cells and cyclophosphamide-induced lung fibrosis. 997 52


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