[Clinical evaluation of quantitative determination of immunoglobulins and complement in ulcerative colitis].
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Adolescent↗
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[Focus groups: preliminary experiences in educational health programs in Brazil].
Since 1989, the public health education section of the University of São Paulo (USP), Brazil, has been using the focus group technique to identify educational problems and evaluate programs being developed. The focus group is a research technique that allows qualitative data to be collected through group sessions involving 6 to 15 persons with some shared trait (for example, sex, age, occupation, role in the community). The group discusses various aspects of a specified subject. This paper describes five research projects in which this technique was used. The projects were carried out by professors in the School of Public Health/USP in the state of São Paulo between 1989 and 1992 with population groups and in health institutions. These experiences showed that the technique is efficient, permitting rapid identification and in-depth analysis of problems from the point of view of the population. Among the drawbacks to the technique is that it uses a small and nonrandom sample, which means that in certain cases focus groups should be considered a complement to quantitative studies. The data described here provide knowledge of perceptions, ideas, opinions, expectations, and social images-in short, the cultural and verbal universe of the population. With this information, educators and administrators can plan and evaluate education programs on the basis of the needs and views of the people they serve, putting into practice the plan to make education more democratic and responsive to the needs of its public.
Adult↗
Quantitative histologic factors for grouping childhood supratentorial neuroglial tumors.
The histologic heterogeneity of childhood supratentorial neuroglial tumors, when quantified, identifies relatively homogeneous subgroups for prognostic purposes and for assignment in clinical trials. Our sample consisted of supratentorial tumors in the Childhood Brain Tumor Consortium. The data consist of reliably identified histologic features and demographic, clinical, operative, and survival information. Factor analysis was used to identify uncorrelated "factors," each represented by a different combination of histologic features in 703 tumors. The defining histologic features were used to label each factor. The heterogeneity of each tumor was summarized using the factor scores for each factor. We compared the survival estimates of subgroups of tumors within common diagnostic classes. We identified five uncorrelated quantitative factors that accounted for much of the histologic variation. Our factor labels were Jumbo, Fibrillary, Proliferative, Spongy, an Oligodendroglial. Two thirds of tumors had high scores on two or more factors, indicating a high degree of heterogeneity among these tumors. Eighty-four percent of supratentorial tumors were accounted for by 19 nonoverlapping relatively homogeneous histologic groups. The five quantitative factors complement standard qualitative taxonomies by summarizing more completely the histologic feature aspects of a tumor than by diagnosis alone and quantify the histologic heterogeneity of individual tumors. Histologically homogeneous groups of tumors are essential for clinical trials, biologic research, and prognostic models.
Adolescent↗
Quantitative histologic factors for grouping childhood infratentorial neuroglial tumors.
We employed factors analysis to quantify the degree of histologic heterogeneity of childhood infratentorial neuroglial tumors. Our data were 26 reliably ascertained histologic features in 1068 children in the Childhood Brain Tumor Consortium database. The factor analysis identified five uncorrelated quantitative "factors," each derived from a different linear combination of the 26 histologic features, that accounted for much of the histologic variation. Histologic features differed in their importance in each factor. The most important features in each factor were used for naming using simple, histologic, familiar descriptive terms: Spongy, Proliferative, Ring, Fibrillary, and Nuclear. Each tumor has a score on each factor. Two-thirds of tumors had high scores for at least two factors, indicating frequent histologic heterogeneity among these tumors. Ninety-five percent of tumors were allocated to 1 of 11 nonoverlapping histologically homogeneous groups. The five quantitative factors complement standard qualitative taxonomies by making explicit the histologic heterogeneity or homogeneity of individual tumors and provide the pathologist with a method that takes advantage of more of the histology of each tumor than conventional nomenclatures. Histologically homogeneous groups of tumors are likely to be of value in clinical trials and biologic research. Prognostic models based on these factors have been published.
Adolescent↗
Production of ultrastructural membrane lesions by the fifth component of complement.
A direct quantitative relationship has been demonstrated between the number of cell bound C4,2 complexes or C5 molecules and the number of ultrastructural lesions visualized on the cell membrane subsequent to immune hemolysis. When bound C4,2 complexes exceeded bound C5 molecules, the number of ultrastructural lesions seen corresponded to the number of C5 molecules. However, in the reverse situation, with bound C5 molecules in excess of bound C4,2 complexes, the latter determined the number of lesions. During the complement-reaction sequence, the lesions first became visible in the nonlytic intermediate complex EAC1,4,2,3,5 and their number was unaffected when lysis was induced by C6-C9. Since the lesions were also demonstrable on the intermediate complex EC5,6,7, it is concluded that the protein C5 is responsible for their production. Once formed, the physical presence of the C5 molecule is no longer required for the manifestation of the lesions as indicated by persistence of lesions after removal of C5 protein by trypsin. The C5-dependent ultra-structural phenomenon has therefore been interpreted to represent a true structural change of the membrane which, however, is not accompanied by a permeability defect.
Animals↗
Complement-dependent cell lysis assay for quantitation of rubella antibodies.
A comparison of the capacity to detect antibody titers to rubella virus was made between the complement-dependent cell lysis (CDCL) assay and the hemagglutination inhibition (HAI) test. Titers detected by CDCL assay correlated with the HAI test. Although the CDCL assay was less convenient to conduct, it proved to be more sensitive than the HAI test.
Antibodies, Viral↗
Quantitative assay of influenza virus soluble antigen by complement-fixation micromethod.
A modified micromethod of complement fixation (CF) reaction for quantitive assay of Influenzavirus soluble (s) antigen was elaborated. The method makes it possible to determine microquantities of s-antigen with an accuracy of +/- 15%. The accuracy of the method was checked by theoretical calculations.
Antigens, Viral↗
[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↗
A simple and sensitive micro-haemagglutination inhibition assay for the semi-quantitation of the third component of complement.
C3-coated cells have been prepared by incubating sensitised erythrocytes with the appropriate dilutions of serum. Inhibition of agglutination of these cells by an anti-C3 serum provides a sensitive and specific assay for C3 detection and semi-quantitation.
Animals↗
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↗
[Alternative complement activation pathway. Its mechanism, importance and quantitative determination].
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Animals↗
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↗
Characterization of kinetics and target proteins for binding of human complement component C3 to the surface-exposed outer membrane of Chlamydia trachomatis serovar L2.
In order to characterize the interaction of human complement with Chlamydia trachomatis, flow cytometry was used to quantitate binding of complement component C3 to elementary bodies of C. trachomatis serovar L2 preincubated in fresh serum in the presence or absence of human polyclonal chlamydial antibody. Isolation of each of the complement activation pathways revealed that C3 was activated most effectively by the alternative pathway. The degree of binding by the classical pathway was proportional to the concentration of antibody, but dual-pathway-mediated binding was not greater than antibody-independent alternative pathway binding. Electrophoresis and immunoblotting of detergent-extracted outer membrane protein-C3b complexes indicated that the chlamydial major outer membrane protein was the primary cell surface moiety binding C3b in both the presence and absence of specific antibody. Hydroxylamine cleavage of outer membrane protein-C3b complexes provided evidence that the majority of C3b is bound to the major outer membrane protein by hydroxyl ester bonds. This result was also unchanged by the presence of specific antibody. An unexpected finding was the apparent binding of anti-C3 antibody to a 40-kDa protein of the chlamydial outer membrane complex, perhaps indicating C3 mimicry on the part of the chlamydial major outer membrane protein.
Amino Acid Sequence↗
Quantitative determination of anti-dsDNA antibodies and antibody/dsDNA stoichiometries in prepared, soluble complement-fixing antibody/dsDNA immune complexes.
We have investigated quantitatively the complement-mediated binding of prepared, soluble 125I-7S IgG antibody/3H-dsDNA immune complexes to human red blood cells (RBCs). We have performed these studies by using a detailed modification of the RBC-CF assay [Pedersen et al., J. Immun. Meth. 38, 269-280 (1980)] which now allows for the simultaneous measurement of both 3H-DNA and 125I-binding to the cells. Our results indicate that, in the case of three SLE patients, their anti-dsDNA antibody titers are sufficiently high that a small fraction of their 125I-7S IgG antibodies (ca 0.1-0.2%) can be identified as specifically anti-dsDNA. We have also used an indirect method (with 125I-labelled rabbit anti-human IgG) for the determination of IgG anti-dsDNA antibodies in complement-fixing antibody/dsDNA immune complexes that bind to RBCs, and the results of these measurements are in reasonable agreement with the direct binding experiments. These studies have also allowed us to estimate the antibody/DNA stoichiometries in complement-fixing immune complexes. The results of these experiments may provide a useful standard for the analysis of monoclonal anti-dsDNA antibodies.
Antibodies, Antinuclear↗