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Pseudothrombocytopenia due to agglutinins.

Pseudothrombocytopenia may have any of a number of causes, one of which is agglutination in vitro. This phenomenon was found in samples of blood from six patients. A serum factor responsible for the agglutination was demonstrated. The factor was dependent upon the presence of EDTA and was more active at room temperature than at 37 C. It could be identified as an IgM immunoglobulin in four cases. In the other two cases definite characterization was not possible, but there was some evidence in favor of an IgM factor. All six patients had elevated serum IgM levels, but they had different and unrelated clinical disorders.

Adolescent↗

[Agglutinins in the diagnosis of pertussis].

The performed investigation showed that the results of agglutination test in diagnostics must be explained with reserve as a supplement to the clinical picture and the other laboratory findings. Titre must be followed at least for 20 days on many blood samples. The more reliable diagnosis according to agglutination test can be given only if we know in question is a nonimmunized child and titre is > or = 1:80. The negative and lower titres cannot exclude or confirm the disease.

Agglutination Tests↗

Presence of serum agglutinins to Campylobacter sputorum subspecies mucosalis in pigs.

Pigs affected with intestinal adenomatosis had agglutinating antibodies to Campylobacter sputorum subspecies mucosalis. The titres obtained varied, depending on the strain used as antigen and appeared to rise with the age of the affected animal. Examination of sera from experimental animals showed that pigs between 30 and 77 days old rarely had antibody, even when they had been exposed to infection. Evidence is also presented indicating that the serological response is specific and that in a limited examination of sera derived from pigs at slaughter almost all animals had high levels of antibody.

Abattoirs↗

Marine biopolymers with cell specificity--III--Agglutinins in the red alga Cystoclonium purpureum: isolation and characterization.

The red alga, Cystoclonium purpurcum, contains agglutinins which selectively agglutinate mouse leukemia cells L5178Y but not L1210. It also agglutinated erythrocytes, lymphocytes, and a marine bacterium. The major component was a glycoprotein having a molecular weight of 12,500 daltons. It contained 5.6% carbohydrates and consisted of two subunits.

Agglutinins↗

Requirement of bromodeoxyuridine for the maintenance of "transformed" characteristics in bromodeoxyuridine dependent cells.

A bromodeoxyuridine(BUdr) dependent cell line, called B4, Which requires BUdr not only for optimal growth but also for the maintenance of the non-contact inhibited state was described previously. We have now shown that contact inhibition in the B4 cells in the absence of BUdr is associated with a marked decrease in the percent of cells synthesizing DNA. The transition to the contact inhibited state in the absence of BUdr does not seem to be due to changes in cyclic AMP levels. It has also been shown that several but not all of the characteristics which distinguish transformed from untransformed cells also distinguish B4 cells grown in the presence of BUdr from B4 cells grown in the absence of BUdr. In addition to being contact inhibited, B4 cells grown in the absence of BUdr have a higher serum requirement, grow less well in soft agar, and are less agglutinable by wheat germ agglutinin than B4 cells grown in the presence of BUdr. Agglutinability by concanavalin A, however, is the same for B4 cells grown in the presence and absence of BUdr. Dependent cells maintained in the presence of BUdr do not form tumors and it is not yet clear how the transformed characteristics of the dependent cells are related to malignancy.

Agglutination Tests↗

Membrane fatty acid replacements and their effect on growth and lectin-induced agglutinability.

The growth of 3T3 and SV101 3T3 cells in a lipid-depleted medium is enhanced by the addition of biotin or some fatty acids. The extent of enhancement depends on the fatty acid(s) supplied. The presence of linoleate is unique, since it induces a morphological alteration in 3T3 cells resulting in a cell similar to an SV101-transformed 3T3 cell. Analyses of the fatty acids from the membrane phosphatides show that the exogenously supplied fatty acids are incorporated and alter the fatty acid composition. This is most clearly evident with heptadecanoate-grown cells, in which this fatty acid and its derivatives comprise over 45% of the fatty acids in the phospholipids. The fatty acid replacements have a striking effect on the temperature dependence of agglutination by wheat germ agglutinin and concanavalin A, implying that fluidity is involved in agglutination. These temperature dependencies and the effect of fatty acid replacements on them were different for the two lectins, but similar for both transformed and untransformed cells. These observations are interpreted as suggesting that the lipid phase is heterogeneous, and that transformed and untransformed cell membranes have regions of similar fluidity.

Agglutination↗

Antigenic structure and relationship between serotypes 1 and 2 of Haemophilus paragallinarum.

Antigenic structure and relationship between serotypes 1 and 2 of Haemophilus paragallinarum were analyzed by the rapid plate agglutination and cross-absorption tests. Encapsulated strain forming iridescent colony type of both serotypes 1 and 2 had at least three antigens: heat-labile and trypsin-sensitive (L), heat-labile and trypsin-resistant (HL), and heat-stable and trypsin-resistant (HS). The L was a major antigen located in a surface and divided serologically into three parts: L1, L2, and L3. The L1 was specifici to serotype 1, the L2 was specific to serotype 2, and the L3 was a common antigen shared by serotypes 1 and 2. The HL and HS were common antigens between serotypes. By trypsinization or heating at 121 C, L antigen lost its agglutinability and agglutinin-producing ability. Nonencapsulated organisms forming noniridescent colony type lacked the L antigen. These results suggested that antigenic structure of H paragallinarum serotypes 1 was L1, L3, HL and HS, while serotype 2 was L2, L3, HL, and HS.

Agglutination Tests↗

Investigations on the in vitro metacyclogenesis of a visceral and a cutaneous human strain of Leishmania infantum.

The in vitro metacyclogenesis of a visceral (VL) and cutaneous (CL) human strain of Leishmania infantum was monitored in order to find out the kinetics of this process and the in vitro infective capacity for macrophages of the metacyclic promastigotes developed. To identify, enumerate, and separate the metacyclic population, the complement-dependent lysis by normal serum and the agglutination by peanut agglutinin (PNA) were used, as they were shown to be useful for the purpose of this study. Maximum percentage of metacyclics was detected by both techniques on the 4th day of growth for VL and the 6th day for CL, and was higher for the VL strain. The in vitro infectivity for macrophages of two strains was assayed, and the high parasitization data obtained were transformed in order to determine the increase of the parasite burden for macrophages throughout the incubation time of the experiments (2-72 h post-infection (p.i.)). This parameter is denominated the infectivity ratio (%I) and calculated as follows: (number of intracellular parasites per infected macrophage at 'x' time p.i./number of intracellular parasites per infected macrophage at 2 h p.i.) x 100. When %I was calculated for promastigotes unagglutinated by PNA (PNA-)--metacyclic or infective promastigotes--at any time of culture, the %I at 72 h p.i. was always much higher than for agglutinated promastigotes (2.1-12.5 times)--non-infective promastigotes--and unfractionated promastigotes from culture (1.7-9.5 times), especially with VL parasites. Likewise, the %I for VL PNA- promastigotes from the 4th day of culture was 1.9 times higher than for CL PNA- promastigotes from the 6th day of culture. The higher resistance to lysis by serum, percentage of metacyclics (PNA-), and infectivity ratio of VL than CL could be related to a higher spreading capability into the host body associated with higher pathogenic effects of the visceral strain than the cutaneous one.

Agglutination Tests↗

Serologic diagnosis in the differentiation of autoimmune hemolytic anemias.

A complex serologic study was carried out in 20 patients with autoimmune hemolytic anemias (AIHA). The methods used were: the direct Coombs' test with specific sera anti IgG, IgM, IgA and complement (C), the direct and indirect test with papain treated erythrocytes (at 37 degrees C), determination of cold agglutinin titer and of warm and cold hemolysins. By these investigations using indigenous sera, the 20 cases of autoimmune hemolytic anemia could be classified: a) according to the thermal behaviour of autoantibodies, into warm antibody AIHA -- 15 cases, cold antibody AIHA -- 2 cases and both warm and cold antibody AIHA -- 3 cases; and b) according to the sensitizing globulin, into: IgG type -- 10 cases, IgG + C type -- 7 cases, IgG + IgM + C type -- 1 case, type C + cold agglutinins -- 2 cases.

Agglutination Tests↗

Allogeneic hemopoietic stem cell transplantation using mouse spleen cells fractionated by lectins: in vitro study of cell fractions.

Mouse spleen cells sequentially agglutinated by soybean agglutinin (SBA) and peanut agglutinin (PNA) previously had been shown to be sufficiently depleted of graft-versus-host activity to allow reconstitution of lethally irradiated allogeneic recipient mice. We have now tested the extent of T-cell depletion in this cell fraction by various in vitro assays, including cytotoxicity testing with anti-Thy-1 antiserum, mitogenic response to the T-cell mitogens concanavalin A and phytohemagglutinin and allogeneic responsiveness in the mixed lymphocytes culture assay. By these criteria the SBA+, PNA+ spleen fraction, used previously in the in vivo experiments, was found to possess about 1% T-cell contamination. The slight contamination with T cells previously found in the singly agglutinated SBA fraction can be removed by a second fractionation with SBA, thus eliminating the possibility that a minor T-cell subpopulation bears receptors for SBA. Finally, we demonstrated that the twice-agglutinated fraction, by SBA and PNA or by SBA alone, contains a significant number of prothymocytes, thereby indicating that mouse prothymocytes bear receptors for both SBA and PNA. The implication of these findings to bone marrow transplantation in humans is discussed.

Animals↗

Microagglutination test for detecting and measuring serum agglutinins of Francisella tularensis.

A microagglutination method utilizing stained antigen for detecting and measuring serum agglutinins against Francisella tularensis is described. The microagglutination and standard tube agglutination techniques were demonstrated to be comparable in sensitivity and specificity. Advantages of the micro method are rapidity and ease of performance, economy of reagents and, in particular, ease of interpreting specific reactivity.

Agglutination Tests↗

[Influence of taliblastin and vinblastine on serum agglutinins determined by 2 methods].

The authors examined the influence of the potential antitumorous preparation taliblastin and the well known cytostatic vinblastin on the formation of serum agglutinins in mice. Two methods were used--the classical reaction of agglutination and the method for inhibition of passive migration of erythrocytes (IPM) described by Schweicer and Gilisen (1974). It was established that vinblastin markedly inhibited the formation of specific agglutinating antibodies, while taliblastin was considerably weaker immunosupresor; The method of IPM was more sensitive than the classical reaction of agglutination and gave a possibility for better determination of immunosuppresive action of cytostatics.

Agglutination Tests↗

Induction of sexual agglutinability by unsaturated fatty acids in Saccharomyces cerevisiae.

Agglutinins in S. cerevisiae are necessary for mating, for recognition between cells of opposite mating type. The mode of agglutinin synthesis is altered by the growth temperature and by the carbon source, from constitutive to inducible synthesis and vice versa. Some of the unsaturated fatty acids tested induced synthesis of agglutinins in cells grown at an elevated temperature, even in the absence of the pheromone. However, synthesis of agglutinins in glycerol-grown cells, that are inducible by the pheromone, was not induced by linolenic acid. Hence, the change in the mechanism of regulation of agglutinability produced by differences in temperature probably differs from that produced by differences in carbon source.

Agglutination↗

A bioserogroup of marine vibrios possessing somatic antigen factors in common with Vibrio cholerae O1.

A bioserogroup of halophilic vibrios, tentatively labelled bioserogroup 1875, strongly agglutinated by O1 antiserum of Vibrio cholerae is described. Cross-agglutination and agglutinin-absorption tests showed that these vibrios had antigens identical with the Ogawa and Inaba factors of V. cholerae O1, although they possessed their own specific antigen distinctive from the A factor that is the specific major antigen of the latter species. In addition, quantitative antigen variation similar to that of the Ogawa-to-Inaba type with V. cholerae O1 was recognized in this halophilic group. They were isolated from estuarine water and are considered to inhabit coastal aquatic environments. Phenotypically, however, this group is not identifiable with any of the species already recognized in the genus Vibrio.

Agglutination Tests↗

Detection of differences in surface carbohydrates of unfertilized and fertilized golden hamster eggs using potato agglutinin.

Changes in the surface carbohydrate moieties of zonae pellucidae and naked vitelli isolated from unfertilized and fertilized hamster eggs were examined through potato agglutinin (STA)-mediated agglutination and binding of FITC-STA as a marker. The results indicated that the stereochemical properties of the saccharide residues within STA-binding sites on the outer surface of the zona pellucida, inner region of the zona pellucida, and the surface of the ooplasmic membrane were different in unfertilized and fertilized eggs, that part of the differences between the two kinds of eggs could be visualized with FITC-STA and that, on the basis of these data, if we use STA as a probe to analyze the surface change following fertilization, we may be able to analyze these changes to a substantial degree.

Agglutinins↗

Pheromone induction of agglutination in Saccharomyces cerevisiae a cells.

a-Agglutinin, the cell surface sexual agglutinin of yeast a cells, was assayed by its ability to bind its complementary agglutinin, alpha-agglutinin. The specific binding of 125I-alpha-agglutinin to a cells treated with the sex pheromone alpha-factor was 2 to 2.5 times that of binding to a cells not treated with alpha-factor. Competition with unlabeled alpha-agglutinin revealed that the increased binding was due to increased cell surface expression of a-agglutinin, with no apparent change in the binding constant. The increase in site number was similar to the increase in cellular agglutinability. Increased expression of a-agglutinin followed the same kinetics as the increase in cellular agglutinability, with a 10-min lag followed by a 15- to 20-min response time. Induction kinetics were similar in cells in phases G1 and G2 of the cell cycle. Maximal expression levels were similar in cells treated with excess pheromone and in cells exposed to pheromone after destruction of constitutively expressed a-agglutinin.

Agglutination↗

Bordetella pertussis culture having only species factor 1.

Eldering, Grace (Michigan Department of Public Health, Grand Rapids), Jack Holwerda, and Julia Baker. Bordetella pertussis culture having only species factor 1. J. Bacteriol. 91:1759-1762. 1966.-Preston's description of Bordetella pertussis 353-Z as a culture possessing only factor 1 was confirmed by agglutination and agglutinin-adsorption tests. The ld(50) values of this culture for mice inoculated by the intracerebral route were approximately the same as that of B. pertussis 18-323, the challenge culture in mouse potency tests of pertussis vaccine. In mouse protection tests, vaccine prepared with the factor 1 culture protected as well against challenge with B. pertussis 18-323, which has a broad antigenic pattern, as did vaccines made with cultures possessing factors 2, 3, 4, and 5 in addition to factor 1. When two groups of similarly immunized mice were challenged with 353-Z and 18-323, respectively, much lower ed(50) values were obtained with the animals challenged with 353-Z. This was true whether a factor 1 vaccine or a vaccine with factors 1, 2, 3, 4, and 5 was tested.

Agglutination↗

Antigenic relationship of the gram-negative organism causing canine abortion to smooth and rough brucellae.

The use of whole-cell antigens in agglutination and agglutinin-absorption tests showed that the organism causing abortion in dogs is similar to rough Brucella abortus, B. melitensis, and B. ovis, but different from smooth Brucella cultures. Water-soluble antigens obtained by ultrasonic treatment and examined by immuno-electrophoresis and gel diffusion show extensive cross-reactions within the genus Brucella, but little or no cross-reaction with similar antigens from other gram-negative genera in the family Brucellaceae. The dog organism showed near identity with rough and smooth Brucella cultures on the basis of immuno-gel diffusion tests with water-soluble antigens, but it lacked the lipopolysaccharide-endotoxin associated with the agglutinogen of smooth brucellae. These findings support the proposal of Carmichael and Bruner for the designation of a new species, "Brucella canis."

Abortion, Veterinary↗