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C Secchi

Publications and source records attributed to C Secchi.

At least 37 records · Page 2Linked to original sources
PubMed1991

High-performance liquid chromatographic determination of D-amino acid oxidase activity.

A new procedure for the assay of D-amino acid oxidase activity has been developed. alpha-Ketoisovaleric acid, derived from D-valine, was estimated by high-performance liquid chromatography after reaction with o-phenylenediamine to give the corresponding quinoxalinol derivative. alpha-Ketovaleric acid was used as an internal standard to ensure the reproducibility of the method. As an example of application, kidney cortex homogenates were analyzed for their D-amino acid oxidase activity. The advantages of the presented procedure for the determination of the enzymatic activity in biological samples compared with previously reported procedures are discussed.

Animals↗
PubMed1991

An enzyme-linked immunosorbent assay for the determination of bovine prolactin in plasma.

Highly purified pituitary bovine prolactin (bPRL) has been used in a sensitive non-competitive enzyme-linked immunosorbent assay of prolactin (PRL) concentrations in plasma. In this assay affinity purified polyclonal antibodies were immobilized to the solid phase in order to capture the antigen, and were also used biotinylated as the detector antibody. The method was found to be reproducible (3% variability between calibration curves) and has been optimized for measuring bPRL concentration in plasma samples, giving an intra-assay coefficient of variation of about 5% and an interassay coefficient of variation of about 9%. The sensitivity of the assay was found to be as low as 0.1 ng/ml of bPRL.

Animals↗
PubMed1991

Binding characteristics of monoclonal antibodies raised against bovine growth hormone.

Monoclonal antibodies have been raised against pituitary bovine growth hormone using the hybridoma procedure. The binding characteristics of the seven selected monoclonal antibodies toward the antigen molecule in its native, chemically or enzymatically treated form have been studied. The reactivities of the monoclonal antibodies with growth hormones from other species and bovine prolactin have also been investigated. The epitopes recognized by four of the produced monoclonal antibodies are conformational, whereas two other monoclonal antibodies bind to sequential determinants. Three antibodies define immunological sites located between residues 6-124 of the bovine growth hormone molecule, and one of this antibody shows higher affinity to human than bovine growth hormone. The immunoreactivity of one monoclonal antibody is enhanced by the previous binding of the antigen to polyclonal antibodies, probably because of a localized conformational change of the bovine growth hormone molecule. This antibody also shows cross-reactivity with all the homologous hormones tested, indicating to recognize a highly conserved antigenic determinant.

Animals↗
PubMed1988

A biotin-avidin sandwich enzyme-linked immunosorbent assay of growth hormone in bovine plasma.

Purified pituitary bovine growth hormone (bGH) has been used to develop a homologous sandwich enzyme-linked immunosorbent assay in which affinity-purified antibodies are immobilized on microtiter plates. Bovine GH bound to specific antibody is then revealed with a second anti-bGH antibody labeled with biotin and peroxidase-conjugated avidin. The method requires only 48 h, including the coating step, and has a sensitivity as low as 0.25 ng/ml of bGH. Statistical analyses (test of parallelism, cross-reactivity among bGH and GH of various species and bovine prolactin, recovery test, within- and between-assay variation, comparison with radioimmunoassay) confirm the high specificity and reproducibility of the method.

Animals↗
PubMed1986

C5a fragment of bovine complement. Purification, bioassays, amino-acid sequence and other structural studies.

C5a and des-Arg-C5a have been purified from bovine serum in milligram amounts. The progress of the purification was followed by measuring the chemotactic activity of the complement fragments. The two polypeptides induce activation of neutrophil-oriented locomotion and secretion with very similar dose/response effects. After preparing a rabbit antiserum to bovine C5a/des-Arg-C5a, a competitive enzyme-linked immunosorbent assay (ELISA) was set up for the detection of C5a from 5 ng/mol to 1 microgram/ml. The complete primary structure of bovine C5a, which consists of 74 amino acids, has been determined by sequence analysis of the tryptic peptides, aligned by peptides derived from a chymotryptic digest, and by partially sequencing the intact molecule. Bovine C5a has a sequence homology of 78% and 70% with porcine and human C5a, respectively, reacts with an antiserum to porcine C5a and is recognized by cell surface receptors on human neutrophils. Finally, the secondary structure of bovine C5a was investigated by circular dicroic spectroscopy and predicted from the amino acid sequence. A comparison of the content and distribution of alpha-helical and/or hydropathic regions, suggests that the three-dimensional structure of C5a might be modeled from the known crystal structure of the homologous C3a molecule.

Amino Acid Sequence↗
PubMed1986

Detection of desamido forms of purified bovine growth hormone.

Tryptic peptide mapping and amino acid sequencing were used for the identification of desamido forms of bovine growth hormone. The major desamido forms of bovine growth hormone resulted from deamidation of asparagine in position 13, 148 and glutamine 140. These forms did not seem to be produced by prolonged treatment of bovine growth hormone in tryptic hydrolysis.

Amino Acid Sequence↗
PubMed1986

Studies on analogues of succinic semialdehyde as substrates for succinate semialdehyde dehydrogenase from rat brain.

The methyl ester of succinic semialdehyde (SSA) was examined as a substrate for succinate semialdehyde dehydrogenase (SSADH) from rat brain. It was found that the ester can be oxidized by the enzyme. Values of Km for SSA-Me were higher than for those for SSA, and for this substrate the enzyme showed a substrate-dependent inhibition. This finding suggests that the carboxylate group of SSA is not essential in the process of inhibition of SSADH by the substrate. Cyclopropyl analogues of SSA, cis- and trans-1-formyl-cyclopropan-2-carboxylic acids, were also individually tested as substrates of SSADH. Only the trans isomer was found to be oxidized to the corresponding dicarboxylic acid; it inhibited the enzyme in the same range of concentrations as SSA. The above data suggest that, as for gamma-aminobutyric acid, SSA is present in an unfolded, transoid conformation at the active site of SSADH.

Aldehyde Oxidoreductases↗
PubMed1986

Thy 1.2 antigen inducing capacity of a calf thymus acid hydrolysate and its fractions.

Splenic null cells of normal mice have been shown to undergo phenotypic induction of Thy 1.2 antigen by various thymic extracts. This experimental model has been employed in characterizing the Thy 1.2 inducing ability of an acid lysate from calf thymus and its various fractions. Incubation of splenocytes from C3H/He mice with unfractionated Thymomodulin induces a dose-response percent increase in the Thy 1.2 antigen bearing cells (9.28 +/- 2.6%, p less than 0.05). Comparable increase appears in fractions 5B (9.46 +/- 0.55%, p less than 0.001) and 5C (8.09 +/- 3%, p less than 0.005), obtained by ultrafiltration and containing peptides of m.w. 600-10,000 d and less than 600 d, respectively. The activity is confined to the acid fraction (6.61 +/- 0.54%, p less than 0.001) isolated by isoelectrofocusing. Control lysate from pig duodenal mucosa was devoid of Thy 1.2 inducing activity. The present findings indicate that Thymomodulin possesses Thy 1.2 inducing capacity on murine null cells and that such activity is maintained and enhanced after various fractionation procedures.

Animals↗
PubMed1986

Stimulation of the antibody response after treatment with thymomodulin in mice immunodepressed with cyclophosphamide and in aging mice.

The capacity of Thymomodulin, a thymic derivative, to restore the antibody response and the number of circulating leucocytes in immunodepressed animals was studied. Administered daily to cyclophosphamide-treated mice, Thymomodulin significantly stimulated the recovery of the antibody response to sheep erythrocytes after a second administration of the antigen. The white blood cell count also showed a rapid and significant increase. Antibody titres were enhanced to normal range values again in immunodepressed rabbits. The possible mechanism of these findings is discussed. Some preliminary results obtained in restoring the immune response in aged mice are also reported.

Aging↗
PubMed1982

Gas chromatographic determination of urinary 3-methoxy-4-hydroxyphenylethyleneglycol sulphate after its ion-pair extraction.

In order to develop a gas chromatographic method with flame ionization detection for the analysis of urinary 3-methoxy-4-hydroxyphenylethyleneglycol sulphate, a new procedure for its isolation has been introduced. The conjugated metabolite has been extracted by an ion-pair technique using tetrabutyl ammonium as counter ion and dichloromethane as organic solvent. Hydrolysis and derivatization of 3-methoxy-4-hydroxyphenylethyleneglycol sulphate have been finally performed by treatment with heptafluorobutyric anhydride. Each step of the procedure, applied to the analysis of urine samples both from normal subjects and from patients with neuroblastoma, has been discussed.

Adult↗