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Results for “quantitative complementation”

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PubMed1986

Micro-complement fixation: a quantitative estimator of protein evolution.

The quantitative immunological technique of micro-complement fixation (MC'F) has been routinely used during the past decade to assess evolutionary relationships among living vertebrate species. The large data base that has been generated, along with the excellent correlations between immunologically measured genetic distances and paleontologically derived estimates of divergence times, have formed the basis for the albumin molecular clock. Immunological distance (ID) involves a logarithmic transformation of experimentally measured antibody concentrations. The justification for this transformation has rested entirely on empirical correlations. Consequently, several other transformations have been proposed as giving better fits to particular data sets. We derive, from first principles, the relationship between ID and the amino acid sequence replacements (AAR) between compared albumins. ID is shown to be a linear estimator of AAR. This ID-AAR relationship is based on a proposed process of antibody assortment and exclusion. We present experimental data confirming that such an antibody assortment-exclusion process occurs in MC'F. This process can explain both the high sensitivity and the quantitative phylogenetic nature of the MC'F assay. The assortment-exclusion process also predicts a divergence limit beyond which MC'F data no longer provide robust phylogenetic data.

Biological Evolution↗

PubMed1978

A quantitative micro-complement fixation method in studies of human wart viruses.

This report offers a description of a quantitative micro-complement fixation method (Cikes, 1975) for detecting human wart virus antigens and their specific antibodies, and proof of the specificity of the reactions being detected. The increased sensitivity demonstrated by chromium-release measurement is compared to the results of visual interpretation of complement fixation.

Antibodies, Viral↗

PubMed2000

[Changes of genomic density polymorphism and quantitative expression of complement receptor type 1 in patients with liver diseases].

OBJECTIVE: To study the changes of genomic density polymorphism and quantitative expression of complement receptor type 1 (CR1) on erythrocytes in patients with liver diseases. METHODS: Polymerase chain reaction (PCR) and Hind III restriction enzyme digestion and the quantitative assay of erythrocytic CR1 were used. RESULTS: The spot mutation rate (25.0% approximately 30.3%) of erythrocyte CR1 density gene in patients with liver diseases was not significantly different with that of healthy individuals (28.0%). The amount of erythrocytic CR1 in patients with liver diseases, except for those with normal liver function, was significantly lower than that of healthy individuals (t=10.44, P<0.0001). The quantitative expression of erythrocytic CR1 in decompensated cirrhosis was obviously lower than that of compensated cirrhosis (t=2.21, P<0.05). CONCLUSION: Defective expression of CR1 in liver diseases is acquired through central and/or peripheral mechanisms. It is very important to study the quantitative expression in the evaluation of the development of liver diseases.

Erythrocytes↗

PubMed1982

Quantitation of rat liver xanthine oxidase by radioimmunoassay. A mechanism for sex-specific differences.

To further delineate the mechanism responsible for the differences in xanthine oxidase activity in male and female Sprague-Dawley rats, a sensitive and specific radioimmunoassay (RIA) was developed for the measurement of hepatic xanthine oxidase. The RIA could detect as little as 5 mg of liver enzyme. Specificity of the RIA was confirmed by 1) Ouchterlony double immuno-diffusion in which a single precipitin band exhibited xanthine oxidase activity, when crude liver homogenate and an enzyme-specific stain were used; 2) parallelism between purified 125I-labeled xanthine oxidase and serial dilutions of crude liver homogenate; 3) a linear correlation between xanthine oxidase activity and the level of enzyme protein; and 4) a single protein band coincident with purified xanthine oxidase, when an immunoprecipitate prepared from antisera and crude liver homogenate was analyzed on sodium dodecyl sulfate (SDS) polyacrylamide gels. Whether xanthine oxidase activity was assayed in the absence of nicotinamide adenine dinucleotide (NAD+) (oxidase form) or in the presence of NAD+ (dehydrogenase), male values were consistently higher, and both forms of the enzyme correlated significantly with each other. When purified to homogeneity, neither form of the enzyme was appreciably affected by 17 beta-estradiol or testosterone propionate. When the RIA was employed, levels of hepatic xanthine oxidase were significantly greater in male than in female rats. We concluded from these data that increased xanthine oxidase activity in the male corresponds to a greater quantitative complement of xanthine oxidase protein. Furthermore, lower xanthine oxidase activity in the female cannot be explained by immunologically cross-reactive material without enzyme activity nor by a direct sex-steroid enzyme interaction.

Animals↗

PubMed2005

Detection of various platelet-associated immunoglobulins by flow cytometry in idiopathic thrombocytopenic purpura.

BACKGROUND: In 1975, Dixson reported that anti-platelet IgG on platelets from patients with idiopathic thrombocytopenic purpura (ITP) is greater than in normal people, by determining anti-platelet antibodies directly on the platelet surface with a quantitative complement lysis-inhibition-assay. Since then, platelet-associated IgG (PAIgG) has been thought of as evidence of ITP. Although platelets from ITP patients show significantly higher PAIgG values than from normal control individuals, PAIgG is not specific for autoantibody because it increases in other than immune ITP patients. METHODS: We analyzed positive platelet percentage with various platelet-associated immunoglobulins: IgG, IgM, IgA, and total immunoglobulins, in the blood from 17 normal donors and 23 ITP patients. RESULTS: The specificity for ITP disease was better in flow cytometry than in ELISA, because, other than ITP, only aplastic anemia was positive in flow cytometry; however, various disorders (aplastic anemia, chronic lymphocytic leukemia, acute myeloid leukemia, and myelodysplastic syndrome) showed positive in ELISA. Flow cytometry methods had the same sensitivity for ITP disease as ELISA. However, it is supposed that there was no nonimmune ITP in this study because the PAIgG negative patients (n = 1) showed positive results in flow cytometry. CONCLUSION: Flow cytometry method was effective for ITP screening, especially for specificity.

Adult↗

PubMed1995

Characterization of DNA polymerase beta mutants with amino acid substitutions located in the C-terminal portion of the enzyme.

We used quantitative complementation assays to characterize individual DNA polymerase beta (Pol beta) mutants for their ability to function in DNA replication and DNA repair. We also describe a screen for detecting mutator activity of DNA polymerase beta mutants. By using these bioassays, together with DNA polymerase activity gels, we characterized 15 new DNA polymerase beta mutants that display a wide spectrum of phenotypes. Most of these mutants are generally defective in their ability to synthesize DNA. However, two of our Pol beta mutants show more complex phenotypes: they are able to function in DNA repair but unable to participate in DNA replication. One of our mutants displays mutator activity in vivo. Our work provides a model to study mutant mammalian enzymes in Escherichia coli with phenotypes that are otherwise difficult to assess.

Animals↗

PubMed1991

Rapid appraisal in an urban setting, an example from the developed world.

Rapid Appraisal has been used as a method to understand communities' own perceptions of their priority needs. It can be used as a tool for formulating joint action plans between communities and service planners and managers. It can also be used to complement quantitative methods of assessing needs by more in-depth qualitative understanding of socio-cultural perspectives. In the example described in this paper Rapid Appraisal is used both as action research and as a qualitative tool in order to define health and social needs of an urban, deprived community in England.

Community Health Services↗

PubMed1998

Can raters consistently evaluate the content of focus groups?

Focus groups are increasingly being used to provide insights to researchers and policy makers. These data complement quantitative approaches to understanding the world. Unfortunately, quantitative and qualitative methodologies have often been viewed as antithetical, rather than complementary, strategies. While focus groups can clearly generate rich information that is unobtainable through other quantitative methods, it is important to determine the degree to which different raters can consistently extract information from transcripts. Thus, our goal was to quantify agreement in the interpretation of transcripts from patient and physician focus groups, using decision-making in ischemic heart disease as a model. We used data from focus groups with both patients and physicians that sought to identify factors affecting diagnostic and treatment decisions in ischemic heart disease. Three raters independently reviewed transcribed audiotapes from focus groups of patients with ischemic heart disease, as well as focus groups of physicians who care for these patients. We found that raters could not distinguish between major and minor factors reliably. More troubling, however, is that consistency regarding the apparently straightforward judgment as to the mere presence or absence of a factor was difficult to achieve. In particular, the three raters of each transcript failed to agree on between one third and one half of the factors. This reasonably high level of disagreement occurred despite the raters: (1) having generated the individual factors themselves based upon their reading a random sample of actual transcripts and (2) being trained in the use of rating forms (including standard definitions of themes). These data suggest that if a single rater evaluates focus group transcripts, as is commonly done, judgments may not be reproducible by other raters. Moreover, a single rater may not extract all important information contained in the transcripts.

Female↗

PubMed2004

Pleiotropic effects of methoprene-tolerant (Met), a gene involved in juvenile hormone metabolism, on life history traits in Drosophila melanogaster.

Life history theory assumes that there are alleles with pleiotropic effects on fitness components. Although quantitative genetic data are often consistent with pleiotropy, there are few explicit examples of pleiotropic loci. The Drosophila melanogaster gene Methoprene-tolerant (Met) may be such a locus. The Met gene product, a putative juvenile hormone receptor, facilitates the action of juvenile hormone (JH) and JH analogs; JH affects many life history traits in arthropods. Here we use quantitative complementation to investigate effects of Met mutant and wildtype alleles on female developmental time, onset of reproduction, and fecundity. Whereas the alleles did not differ in their effects on developmental time, we detected allelic variation for the onset of reproduction and for age-specific fecundity. Alleles influenced phenotypic co-variances among traits (developmental time and onset of reproduction; onset of reproduction and both early and late fecundity; early and late fecundity), suggesting that alleles of Met vary in their pleiotropic effects upon life history. Furthermore, the genetic covariance between developmental time and early fecundity attributed to alleles of Met was negative, indicating consistent pleiotropic effects among alleles on these traits. The allelic effects of Met support genetic models where pleiotropy at genes associated with hormone regulation can contribute to the evolution of life history traits.

Analysis of Variance↗

PubMed1977

Evidence for a common tumour-associated antigen in extracts of human bronchogenic carcinoma.

Xeno-antiserum specific for antigenic components of bronchogenic carcinoma was raised in rabbits, by passively immunizing them to normal human lung antigens at the same time as immunization with a tumour extract from a squamous-cell carcinoma. Antiserum so raised contained minimal quantities of anti-normal antibody which could be removed by a single absorption with glutaral-dehyde-insolubilized normal lung extract. When tested by quantitative complement fixation with a panel of tumour extracts from surgical specimens, it was found that the antiserum gave positive complement fixation with all squamous-cell carcinoma extracts tested, and with some of the extracts from bronchogenic carcinomas of differing pathological types. The antiserum was essentially negative for pooled extracts from normal lung, liver and spleen but gave a weak positive reaction with an extract of pooled foetal lung tissue.

Antibodies, Neoplasm↗

PubMed2003

The genetic architecture of odor-guided behavior in Drosophila: epistasis and the transcriptome.

We combined transcriptional profiling and quantitative genetic analysis to elucidate the genetic architecture of olfactory behavior in Drosophila melanogaster. We applied whole-genome expression analysis to five coisogenic smell-impaired (smi) mutant lines and their control. We used analysis of variance to partition variation in transcript abundance between males and females and between smi genotypes and to determine the genotype-by-sex interaction. A total of 666 genes showed sexual dimorphism in transcript abundance, and 530 genes were coregulated in response to one or more smi mutations, showing considerable epistasis at the level of the transcriptome in response to single mutations. Quantitative complementation tests of mutations at these coregulated genes with the smi mutations showed that in most cases (67%) epistatic interactions for olfactory behavior mirrored epistasis at the level of transcription, thus identifying new candidate genes regulating olfactory behavior.

Animals↗

PubMed1970

Immunochemical measurement of double-stranded RNA of uninfected and arbovirus-infected mammalian cells.

A quantitative complement fixation assay which specifically measures double-stranded RNA has been used to study this RNA extracted from uninfected and arbovirus-infected cells. The double-straned RNA of the uninfected BHK-21 cells sedimented in the 12S region in sucrose gradients. The double-stranded RNA of Sinbis virus-infected cells, as measured immunochemically, included a predominant peak at 12S, a smaller 18S peak, and polydisperse material extending into the 26-30S region. All classes of this RNA detected immunochemically showed a sharp thermal denaturation curve, and increased in amount progressively during infection, with the 12S peak predominant at all times.

Animals↗

PubMed1971

Detection of Australia antigen by radioimmunoassay.

A double-antibody radioimmunoassay has been developed for the detection of Australia antigen and antibodies to Australia antigen in human sera. The assay is simple to perform, and can be completed in 18 hr. Based on parallel studies of human sera, it is approximately 2000 times more sensitive than agar gel diffusion and 200 times more sensitive than quantitative complement fixation. Australia antigen or antibody to Australia antigen was demonstrated in 22 of 23 single serum samples obtained from patients acutely ill with the long-incubation form of hepatitis; eight of the sera were negative when evaluated by less sensitive techniques.

Animals↗

PubMed1985

Interleukin 2 promotes growth and cytolytic activity in human T3+4-8- thymocytes.

Human thymocytes bearing T3 but neither T4 nor T8 antigens (T3+4-8- cells) were obtained after negative selection of thymocytes, either fresh or cultured in medium containing recombinant interleukin 2 (IL-2), by treatment with Na1/34, OKT4A and B9.4 monoclonal antibodies (which recognize T6, T4, and T8 antigens, respectively) and complement. Quantitative flow cytometry showed a 98% pure population of T3+4-8- lymphocytes, which included proliferating cells. The growth and maturation requirements of these thymocytes were characterized and related to the T3-receptor complex and IL-2 pathways, thought to be used by mature lymphocytes. The results show that addition of recombinant IL-2 promotes, in a dose-dependent way, proliferation and acquisition of effector functions by cultured T3+4-8- thymocytes, the growth being inhibitable by monoclonal antibody 33B73 (anti-Tac). Furthermore, cytolytic activity of T3+4-8- cells induced by recombinant IL-2 is specifically blocked by monoclonal antibody OKT3, showing that it operates via the T3-receptor complex and does not require either T4 or T8 molecules. The finding of in vitro responsiveness to recombinant IL-2 in T3+4-8- thymocytes suggests a role of IL-2 in the growth and maturation of cells committed to the T-cell lineage, during intrathymic differentiation, prior to expression of T4 and T8 molecules.

Antibodies, Monoclonal↗

PubMed2005

Ayahuasca in adolescence: Qaualitative results.

Qualitative research was conducted in Brazil among 28 ayahuasca-consuming adolescents members of the União do Vegetal Church, and 28 adolescents who never used ayahuasca. They were compared on a number of qualitative variables, including vignettes measuring moral and ethical concerns. Psychocultural studies utilizing co-occurences of variables in the realm of qualitative studies are useful in understanding and complementing quantitative studies also conducted among this population. Qualitative data show that the teens in the União do Vegetal religion appear to be healthy, thoughtful, considerate and bonded to their families and religious peers. This study examines the modern use of a powerful hallucinogenic compound within a legal religious context, and the youth who participated in these ayahuasca religious ceremonies (usually with parents and other family members) appeared not to differ from their nonayahuasca-using peers. This study helps to elucidate the full range of effects of plant hallucinogenic use within a socially-sanctioned, elder-facilitated and structured religious context.

Adolescent↗

PubMed2026

Exploring the emerging concept of precision rehabilitation: a qualitative study.

PURPOSE: This descriptive qualitative study explored knowledge users' perspectives on precision rehabilitation concepts, barriers, facilitators, and future directions as part of a convergent mixed methods scoping review. MATERIALS AND METHODS: Sixteen clinicians, administrators, and researchers from three North American tertiary care rehabilitation centers were recruited using convenience and snowball sampling to participate in individual semi-structured interviews. Conventional qualitative content analysis followed a deductive thematic approach based on predetermined categories. RESULTS: Analyses revealed three main themes: (1) Although precision rehabilitation shares foundational concepts with precision medicine, there are certain elements, such as personalization, that are uniquely expressed; (2) Rehabilitation-specific facilitators to precision approaches include the use of unobtrusive technology to collect large amounts of data in real-world contexts, while barriers include rehabilitation's typically small, heterogeneous sample sizes; and (3) The future of precision rehabilitation will require collaborative data-sharing to focus on determining care trajectories that enhance functional outcomes. CONCLUSION: Findings provide the first qualitative synthesis of knowledge users perspectives to complement quantitative evidence and inform the emerging field of precision rehabilitation.

Humans↗

PubMed2003

An exposure prevention rating method for intervention needs assessment and effectiveness evaluation.

This article describes a new method for (1) systematically prioritizing needs for intervention on hazardous substance exposures in manufacturing work sites, and (2) evaluating intervention effectiveness. We developed a checklist containing six unique sets of yes/no variables organized in a 2 x 3 matrix of exposure potential versus protection (two columns) at the levels of materials, processes, and human interface (three rows). The three levels correspond to a simplified hierarchy of controls. Each of the six sets of indicator variables was reduced to a high/moderate/low rating. Ratings from the matrix were then combined to generate a single overall exposure prevention rating for each area. Reflecting the hierarchy of controls, material factors were weighted highest, followed by process, and then human interface. The checklist was filled out by an industrial hygienist while conducting a walk-through inspection (N = 131 manufacturing processes/areas in 17 large work sites). One area or process per manufacturing department was assessed and rated. Based on the resulting Exposure Prevention ratings, we concluded that exposures were well controlled in the majority of areas assessed (64% with rating of 1 or 2 on a 6-point scale), that there is some room for improvement in 26 percent of areas (rating of 3 or 4), and that roughly 10 percent of the areas assessed are urgently in need of intervention (rated as 5 or 6). A second hygienist independently assessed a subset of areas to evaluate inter-rater reliability. The reliability of the overall exposure prevention ratings was excellent (weighted kappa = 0.84). The rating scheme has good discriminatory power and reliability and shows promise as a broadly applicable and inexpensive tool for intervention needs assessment and effectiveness evaluation. Validation studies are needed as a next step. This assessment method complements quantitative exposure assessment with an upstream prevention focus.

Adult↗

PubMed1980

Demonstrations of the production of specific antibodies to poly(I).poly(C) in rabbits.

The rabbit antiserum against poly(I).poly(C) purified by hydroxyapatite column chromatography contained three distinct antibodies. They were fractionated into three antibody populations by a series of precipitations (with poly(A).poly(U), poly(I), and poly(I).poly(C)) and their specificities were examined by quantitative complement fixation, double diffusion tests and radioimmunoassay. The first population was common to the double helical structure of double-stranded RNAs. The second was specific for poly(I) and the third was specific for poly(I).poly(C). These studies demonstrated that specific antibodies exclusively reactive with poly(I).poly(C) existed in the rabbit antiserum against poly(I).poly(C).

Animals↗