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Cathodally migrating creatine kinase in sera of patients treated with theophylline, and its diagnostic implications.

Sera from 30 patients, selected only on the basis of having serum theophylline concentrations greater than or equal to 18 mg/L, were assayed for total creatine kinase (CK), isoenzyme CK-MB, and cathodally migrating CK (mCK-2). CK-MB was measured by ion-exchange chromatography. mCK-2 was detected by electrophoresis on agarose gel, with fluorometric scanning. The mCK-2 variant was found in 37% of these 30 patients. None of the patients in the mCK-2 group had CK-MB values exceeding the cutoff value associated with myocardial infarction, so extensive cardiac ischemia was considered unlikely. A review of the patients' records supported this conclusion, and no particular association with any specific disease was found among the mCK-2 group. Investigations with use of goat anti-CK-MM and anti-theophylline indicated that neither CK-MM nor theophylline were present in the mCK-2. Results of specific radioimmunoassay for CK-B subunit were negative. The mobility of the mCK-2 peak was unresponsive to 2-mercaptoethanol, suggesting that CK of mitochondrial origin was not present. These results provide insight into the clinical relevance of mCK-2 and some information concerning its composition.

Adolescent↗

[Cytochrome P450 inactivation on a cathode].

It was concluded on the grounds of spectrophotometric analysis of cytochrome P450 electrolysis products that cytochrome P450 unlike other hem-containing proteins is not reduced on the cathode. At the potential -1.5V it is reduced simultaneously with the transformation into cytochrome P420. Inactivation in the electric field on the electrode surface is irreversible. When the potential is above -1.4V no inactivation of cytochrome P450 takes place.

Chemical Phenomena↗

[Use of a new process of metallic shadowing by cathodic vaporization for scanning electron microscopy of tegumental products of insects].

In scanning electron microscopy, the recent process, not yet much used, of coating subjects with gold by cathodic sputtering, allows one in biology and especially in entomology, to visualize microreliefs, habitually masked, when the old technique called evaporating technique, was used. This process effectively allows one to deposit a very much thinner layer of metal and, because it is performed at room temperature, prevents artifacts caused by the high temperature necessitated by the other technique, whose previous results nevertheless remain generally correct.

Animals↗

Diagnostic evaluation of dot-binding assays for circulating cathodic antigen (CCA) and anti-CCA determinations in schistosomiasis japonica using defined biotinylated conjugates.

Using an affinity purified circulating cathodic antigen (CCA) preparation and an anti-CCA monoclonal IgM antibody, we modified dot-binding assays for anti-CCA and CCA detections with biotinylated conjugates. A study on 3 groups of 138 schistosomiasis japonica patients and 105 healthy individuals demonstrated a high predictive rate of over 95% in both assays, highly comparable to those of circumoral precipitin test (COPT). A blind test in 342 samples of various groups revealed higher detection rates in Ab-binding assay with both acute and chronic case groups (over 90%), but comparatively lower rates in Ag-binding assay with chronic groups (50%-76%). Distinct reductions of either GMRT and dot-indexes were found in 48 praziquantel treated patients whose sera were collected 9 months after chemotherapy. The major target molecules detected by the two binding assays were proved to be the protein incorporated moieties readily precipitated by trichloroacetic acid (TCA), ammonium sulphate and higher concentration of polyethylene glycol (PEG) suggestive of pathological, specific immunoglobulins in the free or complex forms. Non-specific dot reactions were found in some acute and chronic patients with non-relevant bovine serum albumin (BSA) conjugated peroxidase and streptavidin-PO controls, and also in normal sera with the McAb peroxidase conjugate or when working concentration of the biotinylated McAb was not properly titrated. The reliability and proper management of the dot binding assay as an immunodiagnostic tool were discussed.

Animals↗

Detection of IgM antibodies directed against the gut-associated circulating cathodic antigen in sera from Schistosoma mansoni infected patients. Development and comparison of three enzyme-linked immunoassays.

The majority of the human IgM antibodies detected with an immunofluorescence assay (IFA) on adult worms are directed against the gut-associated circulating cathodic antigen (CCA). In order to study this phenomenon further we developed and evaluated three related ELISA methods to specifically detect IgM antibodies against purified CCA. The assays employed: 1) direct coating of CCA, 2) indirect coating of CCA via a monoclonal antibody, and 3) IgM antibody-capture by rabbit anti-mu chain antibodies. Using a group of 46 positive sera, it was found that the three ELISA's and the IFA were significantly correlated. To discriminate between positive and negative sera we used a cut-off level of average reactivity + 3 standard deviations of 50 negative sera. False negative reactions were not found in any of the ELISA's, while both in the direct and indirect ELISA one false positive reaction occurred. For further studies or diagnostic use the antibody-capture ELISA is recommended.

Animals↗

[The cathode bound group antigen of dysentery-provoking escherichieae (author's transl)].

Antigens from disrupted cells of dysentery-provoking and of non-enteropathogenic Escherichieae were submitted to immunoelectrophoresis on cellulose acetate stripes at pH 8.0. Among 6 immune sera produced for this purpose by immunizing rabbits against desintegrated dysentery bacteria, only one contained a precipitine reacting with an antigen similar to the "generic antigen" of BELAYA. This - at pH 8.0 - cathode-bound group antigen (KGA) could not only be found in virulent but also in 5 attenuated cultures and in 5 from 6 avirulent strains of several dysentery types. Only the - apathogenic - type culture 1111/55 of dysentery-provoking E. coli O 136 showed no KGA-reaction. Some sources of methodical errors responsible for false outcomes of immunopherogrammes have been discussed.

Antigens, Bacterial↗

Schistosoma mansoni excretory circulating cathodic antigen shares Lewis-x epitopes with a human granulocyte surface antigen and evokes host antibodies mediating complement-dependent lysis of granulocytes.

Parasitic worms of the genus Schistosoma excrete relatively large amounts of immunogenic glycoproteins (circulating cathodic antigen [CCA]) that contain polysaccharide side chains with the trisaccharide Lewis-x (L(ex)) as a repeating unit. These carbohydrates evoke high titers of specific IgM antibodies that cross-react with the repeating L(ex) units on the surface of granulocytes. Consequently this might lead, in the presence of complement, to lysis of the granulocytes. In the present study, this hypothesis was investigated using anti-CCA mouse monoclonal antibodies (MoAbs) and polyclonal antibodies purified from sera of infected humans. By flow cytometry, it was demonstrated that the mouse MoAbs directed against CCA strongly recognized the granulocytes. It could also be shown that these MoAbs, as well as anti-CCA IgM antibodies purified from infected human sera, caused lysis of granulocytes in a complement-dependent cytotoxicity assay. Sera from healthy controls or from patients with other helminth infections resulted in negligible granulocytotoxicity. These in vitro observed phenomena may explain the mild to moderate neutropenia that occurs in schistosomiasis patients.

Animals↗

Ketoprofen tissue permeation in swine following cathodic iontophoresis.

BACKGROUND AND PURPOSE: Pharmacokinetic assessment of drug tissue permeation following iontophoresis is limited. The depth of ketoprofen tissue permeation following cathodic iontophoresis (4 mA, 40 minutes) and the stereoselectivity of drug delivery were examined in this study. SUBJECTS: Ketoprofen (750 mg) was iontophoresed onto one porcine medial thigh, with passive drug permeation conducted on the other thigh. METHODS: Skin, subcutaneous fascia, and muscle biopsies from the drug delivery sites were harvested and stored separately, and the "R" and "S" ketoprofen enantiomers were determined. Results. Iontophoretic and passive applications yielded equivalent total ketoprofen concentrations in the skin and fascia. In contrast, multivariate analysis demonstrated that the ketoprofen concentration in the first centimeter of muscle following iontophoresis was greater than the drug concentration in the deeper underlying muscle layers and greater than that delivered to any muscle layer following passive delivery. No transcutaneous stereoselective delivery) of ketoprofen was detected. CONCLUSION AND DISCUSSION: Compared with passive delivery, iontophoresis enhances nonstereoselective ketoprofen permeation into the fascia-muscle interface. With delivery to deeper tissue sites, however, there is no apparent enhancement over passive application.

Animals↗

Cathodic stripping voltammetry of nanogram amounts of selenium in biological material.

A method is described to determine selenium in biological material, based on cathodic stripping voltammetry. Following wet ashing, the selenium was extracted into benzene as the 3',4'-diaminophenylpiazselenol. The selenium was subsequently back-extracted into dilute acid for analysis. Analyses of NBS Bovine Liver demonstrated that the method was capable of recovering 96+/-9% of the selenium present. The detection limit and working range were 3 ng/g and 0-10,000 ng/g, respectively. The method was also applied to the determination of selenium in rapeseed oils and seed.

Animals↗