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Preliminary note on the antipsychotic efficacy of bromperidol.

Bromperidol was administered to 20 acute and chronic psychotic inpatients during six to eight weeks at a mean daily dose of 18 to 10 mg. The neuroleptic was given orally or intramuscularly once a day (in the morning in order to evaluate its therapeutic and side effects). In 13 patients, bromperidol was substituted for haloperidol; in 7 cases, it was given immediately after the washout period. Descriptive psychopathology was complemented with quantitative psychopathology (ad hoc 29-item scale, AMDP Psychopathology Scale, Bf/Bf' Mood Scales). According to a preliminary analysis of data, both descriptive and psychometric, bromperidol seems to be a potent antipsychotic drug without disturbing adrenolytic side-effects or sedation, with classical extrapyramidal effects of the hypertonia type.

Adult

Expression of HLA-B27 antigens on mononuclear leucocytes in ankylosing spondylitis.

Differences in expression of HLA-B27 antigens on immune competent cells might play a role in the susceptibility to environmental factors which may be responsible for the initiation of ankylosing spondylitis (AS). Using a quantitative complement-mediated lymphocytotoxity assay we determined the expression of HLA-B27 antigens on the membranes of mononuclear cells obtained from 20 patients with AS, four patients with other seronegative arthropathies and eight healthy controls. The variation in expression of B27 between individuals was quite extensive, but there was no significant difference in the mean titration curves obtained for each of the three groups. These findings suggest that the expression of HLA-B27 antigens on the membrane of mononuclear leucocytes does not play a role in the pathogenesis of AS.

Antigens, Surface

Quantitative relationships of the fourth complement component in human cerebrospinal fluid.

A technique for the measurement of cerebrospinal fluid C4 concentration in unconcentrated specimens has been developed with the methods of electroimmunodiffusion and immunofixation. The method has proved to be reproducible and requires only microliter volumes of undiluted cerebrospinal fluid (CSF). The mean value for CSF C4 concentrations in 16 neurologically normal individuals was 325 +/- 32 mug/100 ml. A positive correlation between CSF C4 concentration and the concentration of CSF albumin and total protein was observed. The positive correlation between the concentrations of CSF C4 and albumin was, however, more clearly defined than the relationship of CSF C4 to total CSF protein.

Albumins

[Current overview of diagnostic possibilities of complement analysis].

The complement system is composed of at least 21 serum proteins and 8 cell surface receptors sensitive to complement components or their fragments. Qualitative and quantitative analysis of complement components in clinical samples has become standard procedure in many laboratories thanks to the availability of easy-to-perform test kits. However, interpretation of results is still a task that requires great skill and the situation are rare, in which analysis of the complement system goes beyond diagnosis into therapeutic consequences. The present paper is an update of previous similar reviews now available in the international literature; in addition, the author attempts at bringing the often complicated world of complement and clinical practice together and thus contributes to answering the question: what should the clinician know about complement?

Antigen-Antibody Complex

Anti-peripheral myelin antibody in patients with demyelinating neuropathy: quantitative and kinetic determination of serum antibody by complement component 1 fixation.

The role of anti-peripheral nerve myelin antibody (anti-PNM Ab) in the pathogenesis of acquired demyelination of peripheral nerve is unclear, in part, due to the poor correlation between antibody and disease activity. Previous studies show that only 27-50% of patients with acute demyelinating neuropathy or Guillain-Barré syndrome (GBS) had serum Abs to peripheral nerve or PNM as demonstrated by consumption of hemolytic activity of serum complement 1 (C1) fixation and transfer assay, quantitative determinations of anti-PNM Ab showed significantly high titers in the serum of patients with GBS, chronic and recurrent polyneuritis, and paraproteinemia associated with peripheral neuropathy. All 11 patients with acute-phase GBS had Ab titers 6-56 times higher than controls. In 6 GBS patients, serial Ab determinations showed that titers were highest on admission, fell rapidly the first week, and became undetectable or barely detectable by the third week. Declining Ab titers coincided with cessation of clinical progression. In 3 GBS patients, depletion of serum IgM lowered anti-PNM Ab titers significantly, whereas IgG depletion failed to produce a similar effect. This study shows that the C1 fixation and transfer assay is a sensitive method to detect anti-PNM Ab in the serum of patients with a variety of demyelinating neuropathies and provides good correlation between Ab level and the clinical course of GBS patients. It may provide important information about the pathogenesis of the demyelinating neuropathies.

Antibodies

Double fluorescence technique for measurement of complement-fixing antibody to lymphocyte subsets.

A double fluorescent antibody method for quantitating human complement-fixing antibody to lymphocyte subclasses has been developed. The indicators in this system are a C6-deficient serum as a non-lytic source of complement, rhodamine-labeled anti-C3 and fluorescein-labeled murine monoclonal antibodies to human lymphocyte subsets. The basic procedure is to incubate lymphocytes with the unknown serum and then to add C6-deficient serum. The binding of C3 is indicated by staining with rhodamine-labeled anti-C3 and the subset class of the lymphocyte so stained is determined by binding of fluorescein-tagged anti-OKT4 or -OKT8 antibodies. The occurrence of both red and green cell surface fluorescence denotes the presence of a complement-binding antibody to the lymphocyte subset defined by the monoclonal antibody. In addition to defining the specificity of complement-fixing anti-lymphocyte antibodies, this technique is more sensitive than the microcytotoxicity assay.

Acquired Immunodeficiency Syndrome

Quantitation of C3 nephritic factor of alternative complement pathway by an enzyme-linked immunosorbent assay.

We have developed an enzyme-linked immunosorbent assay (ELISA) for the quantitation of C3 nephritic factor of the alternative pathway of complement (NeFA). Incubation of the NeFA-positive serum (patient KS serum) with normal human serum (NHS) in Mg-EGTA resulted in the formation of C3-B-IgG complex. No complex was formed in EDTA. At first this was detected as three types of complexes: C3-IgG, B-IgG and B-C3, by the combination of antibodies. The reaction mixture in Mg-EGTA was filtered through an ACA 22 column, from which the complexes were eluted in the same part as the first protein peak. When IgG purified from KS serum was incubated with NHS in Mg-EGTA, B-C3 complex increased in proportion to the dose of IgG. These results indicated that only one kind of complex consisting of IgG, C3 and B (IgG-C3-B) was generated by the addition of NeFA to NHS. Serum NeFA could be quantified as the titer of B-C3 complex formed after its incubation with NHS in Mg-EGTA. Using the ELISA method, NeFA was positive in five out of six patients with membranoproliferative glomerulonephritis (MPGN) type II and in only one of 17 with MPGN type I. Titers obtained by the new method were in good accordance with those by C3 conversion and C3bBb stabilization assays for NeFA, and the new method was more exact and simple than the conventional methods.

Antigen-Antibody Complex

Effect of amnionitis on the complement system of preterm infants.

The development of the complement system was studied by quantitation of total hemolytic complement activity (CH50), C1q, C3, C4, and C3 split product (C3d) in cord plasma of nine human fetuses (17-22 weeks of gestation), 110 preterm (24-36 weeks of gestation) and 30 term neonates. The complement levels were analyzed in relation to various illnesses of preterm infants. Histological examination of the placenta revealed a higher incidence of amnionitis in the placenta of less than 34 gestational weeks. In cases without amnionitis, there were significant correlations between complement levels and gestational age. In cases with amnionitis, the complement system was activated even in infants of less than 28 weeks gestation. The complement levels correlated with the extent of the inflammation in the placentas and umbilical cords except for C1q. In infants with Wilson-Mikity syndrome, complement levels other than C1q were significantly elevated in comparison with those of infants with respiratory distress syndrome. In the group of preterm infants without amnionitis, no differences were found between infants with intrauterine growth retardation and those with growth appropriate for gestational age.

Chorioamnionitis

Serum immunoglobulin and complement levels in patients with psoriasis vulgaris.

In 102 patients with psoriasis vulgaris, divided into group with slight psoriatic form and the group with severe psoriatic form, the serum immunoglobulin and complement levels were determined. The quantitation of immunglobulin and complement was carried out by single radial immundiffusion. It was found that the IgM levels were significantly decreased in both investigated groups. The IgA and IgG levels were significantly increased only in patients with severe psoriatic form. While the beta1A (C'3) levels were significantly increased in both investigated groups the beta1E (C') levels were not significantly changed compared to control group. It is concluded that the immunoglobulin imbalance in patients with psoriasis, opposite to complement, is depended on the severity of clinical form.

Adolescent

Critical role of the conversion of the third complement component C3 (beta 1C/beta 1A) for its immunochemical quantitation.

For the immunochemical quantitation of the third component of complement C3 the state of conversion of this protein in the sample and the reference serum has to be guarded. Antisera raised against C3 may show different antibody specificity for the antigenic pattern of the C3 proteins which change during proteolysis by activation and also by ageing. It is shown that anti-C3-sera gave differences from -17% to + 91% comparing fresh and aged aliquots of the same sera directly on the same immunodiffusion plate corresponding to a particular anti-C3-serum. The use of three different monospecific antisera directed against the A determinant of C3 seemed to give results with good correlation in respect to C3 conversion. Glass and polystyrene plastic tubes did not have a significantly different effect on the C3 values in respect to C3 conversion. The in vitro acceleration of C3 conversion by thrombin or an inulin-like polysaccharide did not give consistent results in this context.

Complement C3

Analysis of genetic complementation by whole-cell microtechniques in fibroblast heterokaryons.

A whole-cell microtechnique for the determination of complementation of human metabolic disorders is presented. This procedure permits the isolation of individual multinucleate cells produced by cell fusion for the quantitative evaluation of complementation. Mutant fibroblasts with a deficiency of propionyl-CoA carboxylase activity (EC 6.4.1.3) that had been mapped to complementation groups pcc and bio were used to evaluate the microtechnique. Complementation was monitored by the determination of [14C]propionate incorporation into cellular macromolecules. Single cells or a small number of cells were isolated from plastic film dishes after radioactive incubation by cutting out the portion of the plastic film holding the desired cells. Isotope incorporation was linear in 10-50 unfused cells and in 10-50 fused normal cells containing five or more nuclei. There was also a direct correlation between the nuclear content of cells and the amount of isotope incorporated. Three pcc and two bio mutants were fused in pairwise combinations by means of polyethylene glycol and complementation was determined by isotope incorporation in sets of 50 multinucleate cells, each cell isolated individually. The results agreed with autoradiographic data for both complementing and noncomplementing strains. The method is quantitative and gives severalfold higher sensitivity than current procedures. The method can be applied to the complementation analysis of a wide variety of inherited disorders of intermediary metabolism.

Acyl Coenzyme A

Complement in cystic fibrosis.

Quantitative and functional assessments were made of both the classical and alternative pathways of complement activation in sera from 23 patients with cystic fibrosis. The classical pathway functioned similarly in patients and controls as measured by CH50 titre. Alternative pathway function, initiated in patient sera by incubation with inulin, was equal to that of controls as determined by cleavage of Factor B and C3, and by the consumption of terminal components. Factor B, however, was more readily activated in patient than in control sera. This rapid alteration of Factor B did not lead to accelerated or more extensive activation of the terminal complement components via the alternative pathway when assessed by C3 cleavage and the consumption of terminal components. Thus, a complement deficiency was not found. The importance of the easily activated Factor B is undefined.

Adolescent

Epitope specificities and quantitative and serologic aspects of monoclonal complement (C3c and C3d) antibodies.

Twenty-two monoclonal antibodies to human C3c and ten to C3d were obtained by hybridization after the immunization of mice with complement-coated human red cells and/or purified human complement components. C3c antibodies were variable in their agglutination reactions with cells coated with C3 by antibody in vitro; more consistent and potent reactions with these cells were observed with anti-C3d, and all anti-C3d reacted with red cells coated with C3 in vivo. Immunoradiometric assays were used to estimate antibody concentration, affinity, and epitope specificity. The antibody content in ascitic fluids varied from less than 0.1 mg per ml to 5.6 mg per ml. The estimated values of antibody affinities for Sepharose-coupled C3 ranged from 2.8 X 10(6) l per M to 5.0 X 10(8) l per M; on average, IgM antibodies had higher affinities than IgG antibodies. Competitive binding assays showed that the monoclonal antibodies recognized at least seven different epitopes, four on the C3c and three on the C3d fragment of C3. When the results of serologic and quantitative assays were compared, no convincing relationship was found between serologic performance and epitope specificity, antibody concentration, or affinity. IgM antibodies generally gave higher agglutination scores than IgG antibodies, and Ig class was the only useful predictor of serologic efficacy.

Animals

Complement-dependent cytotoxic antitumor antibody. II. Quantitative determination of cell-bound immunoglobulin M1.

A method for the quantitative determination of rabbit IgM bound to cell surfaces has been developed. This method was based on the ability of goat IgG specific for rabbit IgM heavy chains to bind 125I-labeled protein A when bound to the antigen. With the use of this technique the production of specific IgM antitumor antibodies in New Zealand White rabbits after immunization with guinea pig hepatoma cells line-1 and line-10 was followed. Differences in the production of IgM were observed between the different bleedings from rabbits immunized with line-1. No significant IgM antibody was produced following immunization of rabbits with line-10 tumor cells. This indirect method for determining IgM on the cell surfaces was objective, easy to perform, and detected complement-fixing and noncomplement-fixing antibodies. In addition, this technique could be applied to quantify other components on the cell surface for which a suitable specific antibody was available.

Animals