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PubMed1981

Growth and surface properties of dispase dissociated human fibroblasts.

The dissociation of human fibroblast cultures with the bacterial neutral protease (Dispase II) was monitored by viability and growth measurement and was compared to the effect of trypsin and EDTA. Cell suspensions with high viability and excellent growth were obtained after 10 min incubation at 37 degrees C in 4 U/ml dispase in 0.02% EDTA. A two to three-fold increase in mitotic index occurred in the cultures within 48 h after dispase dissociation. The initial rate of aggregation was comparable to that of trypsin or EDTA dissociated cells, but attachment to a substratum and agglutination by Wheat Germ Agglutinin were markedly enhanced. The results indicate that dispase-EDTA provides a valuable alternative to the enzymatic dissociation with trypsin. Moreover, it is an additional tool for the dissociation of cultured cells and for the study of the surface properties of single cells.

Cell Adhesion↗
PubMedDate not supplied

Blood typing of blood stains on sweat stains by the isoagglutinins detection method.

When a blood typing is made for mixed stains of sweat and blood, erroneous results may be obtained. The reason is that the blood group substance in the sweat is detected at the same time as that in the blood. In this paper the typing of the blood stain on the sweat stain is carried out by the detection of isoagglutinins which may give additional information to the forensic serologist.

ABO Blood-Group System↗
PubMed1977

Sperm/cervical-mucus crossed hostility testing and antisperm antibodies in the husband.

The behaviour of sperms has been investigated in preovulatory cervical mucus in 44 infertile couples. In 22 couples, immobilising and agglutinating autoantibodies were detected in the husband's sera in high titres. In 10 couples, antisperm antibodies were detected in the husbands by indirect immunofluorescent testing. In 12 couples, no evidence of antisperm antibodies was found in either husbands or wives. The results obtained with husband and wife were compared with the behaviour of the husband's sperms in cervical mucus from fertile donors, and with the behaviour of sperms from fertile donors in the wives' mucus. This crossed hostility test indicated that high tires of immobilising and agglutinating antisperm antibodies in the husband effectively prevented the sperms from penetrating the cervical mucus, even though the sperms appeared normal on seminal analysis. Antibodies detected by indirect immunofluorescence did not have this effect. Poor penetration was also observed with low sperm motility or poor cervical mucus. It is concluded that this test, taken with the postcoital test, could provide a useful screen for immunological causes of infertility and an accurate test for the clinical relevance of antisperm antibody tests.

Agglutination Tests↗
PubMed1983

Comparison of standard tube and microagglutination techniques for determining Brucella antibodies.

A microagglutination method for determining the agglutinating and "blocking' antibodies to Brucella abortus is described. A collection of sera from healthy blood donors in two rural areas of New Zealand were tested by the microagglutination methods and the standard methods in tube. The results are compared and show that where discrepancies occur, these are due to the microagglutination methods being more sensitive. It is concluded that these are suitable methods for screening populations.

Agglutination Tests↗
PubMed2001

Persistence of anti-leptospiral IgM, IgG and agglutinating antibodies in patients presenting with acute febrile illness in Barbados 1979-1989.

The persistence of anti-leptospiral IgM and IgG antibodies and agglutinating antibodies was studied in serologically confirmed cases of severe leptospirosis during the acute illness and over periods of several years after recovery. The antibody response in non-leptospirosis patients presenting to hospital with similar symptoms over the same period of time was used to estimate the background antibody level to leptospirosis in the community. All patients enrolled in the study had blood samples collected twice in the acute stage of illness, once during convalescence and then annually from the time of initial hospitalisation until the end of the study period. Six hundred and thirty-eight patients presented to hospital with acute febrile illness, of whom 321 were diagnosed with leptospirosis. Patients who had severe leptospirosis commonly remained seropositive, with IgM, IgG and agglutinating antibodies detectable for several years after infection. A significant proportion of cases had high titres of agglutinating antibody detectable by the microscopic agglutination test (> or = 800). There were marked differences in the magnitude and duration of persistence of agglutinating antibodies directed against different serogroups. More than 20% of cases with evidence of infection with serogroup Autumnalis retained titres of >800, 4 years after the acute illness. In one case a titre of 800 was detected 11 years after infection. Persistence of agglutinating antibody titres can create problems in interpretation of serological results and make it impossible to estimate the time of infection, given a specific titre. This study demonstrates that in endemic areas where seroprevalence is high, use of a single elevated titre is not reliable to define a current infection.

Agglutination Tests↗
PubMedDate not supplied

Differentiation of granulocyte antigens demonstrated by granuloagglutinins and granulocytotoxins.

Twenty sera containing granuloagglutinins along with granulocytotoxins were typed with 27 to 54 samples of granulocytes different donors. For the most part, positive results of the microgranuloagglutination test and microgranulocytotoxic test were not in agreement. The positivity of the agglutination test was more frequent (35.24%) than that of the cytotoxic test (18.9%). The statistical evaluation of the results for each serum in both test made by calculating the correlation coefficient showed the two types of antibodies to be independent, and to determine different antigens.

Agglutination Tests↗
PubMedDate not supplied

Granulocyte specificities detected by cytotoxicity and agglutination.

In a sample of 100 individuals from the Prague population antigenic specificities of granulocytes were determined on the basis of agglutination activity by means of standard typing sera (Bethesda, MD). The specificities NA1, NA2, NB1 were detected in 59, 87, and 90 percent with gene frequency of 0.360, 0.640, and 0.684, respectively. One serum from a pregnant woman reacted with 76 percent of persons up to the titer 8 or 16. When correlating this serum with N typing sera, the best match was found for anti-NA2 serum (p less than or equal to 0.01). On the basis of cytotoxic activity, using specific granulocytotoxic sera obtained by screening sera from pregnant women and multitransfused patients, two antigenic specificities were detected. The incidence of G1 and G2 was 55 and 67 percent and the gene frequency 0.555 and 0.455, respectively. The comparison of specificities based both on agglutination and cytotoxic activity showed a significant association of G1 with NA2 and G2 with NA1 (p less than or equal to 0.001).

Agglutination Tests↗
PubMed1973

Ganglioside patterns and phenotypic characteristics in a normal variant and a transformed back variant of a simian virus 40-induced hamster tumor cell line.

Ganglioside patterns of a cloned Simian virus 40- (SV40) induced hamster tumor cell (Cl(2)TSV(5)-S), its normal variant (Cl(2)TSV(5)-R) which are Cl(2)TSV(5)-S gradually adapted to grow in the presence of 2 microg/ml actinomycin D and exhibit certain normal phenotypic characteristics, and its back variant (Cl(2)TSV(5)-RR), which are Cl(2)TSV(5)-R cells grown in the absence of actinomycin D for more than 60 passages and which exhibit greater phenotypic similarity to Cl(2)TSV(5)-S cells, have been analyzed. All three cell lines contain N(acetylneuraminyl) galactosylglycosyl ceramide (hematoside, GM(3)), N-acetylgalactosaminyl (N-acetylneuraminyl) galactosylglucosyl ceramide (GM(2)), and a higher ganglioside tentatively identified as disialohematoside. However, Cl(2)TSV(5)-R have more GM(2) than Cl(2)TSV(5)-S whereas Cl(2)TSV(5)-RR contain an intermediate amount of GM(2). The amount of GM(2) is correlative with the activity of UDP-N-acetylgalactosamine: hematoside N-acetylgalactosaminyl transferase in the extract of the three cell lines and with their agglutination by wheat germ agglutinin.

Animals↗
PubMed1962

Escherichia coli associated with colostrum-free neonatal pigs raised in isolation.

Escherichia coli 08 was the most frequent coliform isolated from the blood and liver of morbid and dead neonatal, colostrum-free piglets raised under extremely sanitary conditions. This strain accounted for 67 per cent of the typable E. coli. The next most numerous strain occurred at a frequency of 6 per cent. Hence, E. coli 08 was considered the main coli enteropathogen in our experimental, isolated environment. In random samples of the feces of healthy and diarrhetic neonatal piglets, 24 per cent of the typable E. coli was type 08. When a directed effort was made to isolate E. coli 08 from the feces of neonatal piglets in a healthy, colostrum-free litter, this strain was isolated from 17 per cent of the total E. coli colonies examined. Thus, the enteropathogen E. coli 08 was ubiquitous in the feces of piglets in our environment, making up approximately 20 per cent of the fecal E. coli. 85 per cent of the bacteremia and death in which E. coli was isolated from blood or liver occurred in piglets fed diets void in bovine and porcine gamma globulin. Tube agglutination tests demonstrated that agglutinins to E. coli 08, and other serotypes as well, were present in bovine colostrum and to a lesser extent in porcine colostrum. These agglutinins were practically lacking in solutions of porcine and bovine gamma globulin. Feeding 10(9)E. coli 08 bacteria to 2-week-old, colostrum-free, gamma globulin-free, 08 agglutinin-free piglets did not produce visible disease.

Animals↗
PubMedDate not supplied

Genetic analysis of the aggA locus involved in agglutination and adherence of Pseudomonas putida, a beneficial fluorescent pseudomonad.

An isolate of Pseudomonas putida, which rapidly adheres to plant roots, is agglutinated by a glycoprotein from root surfaces. Agglutination is prevented and adherence to the root surface is diminished by Tn5 insertion in mutant 5123. Two cosmid clones from wild type P. putida and a 2.7-kbp EcoRI-HindIII subclone present in both cosmid clones restored agglutinable to wild type levels in transconjugants of the nonagglutinable (Agg-) Tn5 mutant 5123. These three clones increased agglutinability in transconjugants of the parental Agg+ isolate. The 2.7-kbp EcoRI-HindIII subclone restored adherence to bean root surfaces of 5123 to wild type levels in a short-term binding assay. Deletion analysis of the 2.7-kbp fragment indicated only 1.45 kbp was necessary for complementation of agglutinability in 5123. This sequence, termed the aggA locus, contains an open reading frame of 1,356 nucleotides encoding a predicted 50,509-Da protein. The distribution of the aggA locus in plant-associated bacteria, as detected through Southern hybridization, is limited to bacteria that express the agglutination phenotype.

Agglutination Tests↗