Complete Agglutination Research Articles

Micro-Droplet Platform for Exploring the Mechanism of Mixed Field Agglutination in B3 Subtype.

B3 is the most common subtype of blood group B in the Taiwanese population, and most of the B3 individuals in the Taiwanese population have the IVS3 + 5 G > A (rs55852701) gene variation. Additionally, a typical mixed field agglutination is observed when the B3 subtype is tested with anti-B antibody or anti-AB antibody. The molecular biology of the gene variation in the B3 subtype has been identified, however, the mechanism of the mixed field agglutination caused by the type B3 blood samples is still unclear. Therefore, the purpose of this study was to understand the reason for the mixed field agglutination caused by B3. A micro-droplet platform was used to observe the agglutination of type B and type B3 blood samples in different blood sample concentrations, antibody concentrations, and at reaction times. We found that the agglutination reaction in every droplet slowed down with an increase in the dilution ratio of blood sample and antibody, whether type B blood or type B3 blood was used. However, as the reaction time increased, the complete agglutination in the droplet was seen in type B blood, while the mixed field agglutination still occurred in B3 within 1 min. In addition, the degree of agglutination was similar in each droplet, which showed high reproducibility. As a result, we inferred that there are two types of cells in the B3 subtype that simultaneously create a mixed field agglutination, rather than each red blood cell carrying a small amount of antigen, resulting in less agglutination.

Contraceptive efficacy of sperm agglutinating factor from Staphylococcus warneri, isolated from the cervix of a woman with inexplicable infertility

BackgroundVoluntary control of fertility is of paramount importance to the modern society. But since the contraceptive methods available for women have their limitations such as urinary tract infections, allergies, cervical erosion and discomfort, a desperate need exists to develop safe methods. Vaginal contraceptives may be the answer to this problem, as these are the oldest ways of fertility regulation, practiced over the centuries. With minimal systemic involvement, these are also the safest. Natural substances blocking or impairing the sperm motility offer as valuable non-cytotoxic vaginal contraceptives. Antimicrobial peptides (AMPs) isolated from plants, animals and microorganisms are known to possess sperm immobilizing and spermicidal properties. Following this, in the quest for alternative means, we have cloned, over expressed and purified the recombinant sperm agglutinating factor (SAF) from Staphylococcus warneri, isolated from the cervix of a woman with unexplained infertility.MethodsGenomic library of Staphylococcus warneri was generated in Escherichia coli using pSMART vector and screened for sperm agglutinating factor (SAF). The insert in sperm agglutinating transformant was sequenced and was found to express ribonucleotide-diphosphate reductase-α sub unit. The ORF was sub-cloned in pET28a vector, expressed and purified. The effect of rSAF on motility, viability, morphology, Mg++-dependent ATPase activity and acrosome status of human sperms was analyzed in vitro and contraceptive efficacy was evaluated in vivo in female BALB/c mice.ResultsThe 80 kDa rSAF showed complete sperm agglutination, inhibited its Mg2+-ATPase activity, caused premature sperm acrosomal loss in vitro and mimicked the pattern in vivo showing 100% contraception in BALB/c mice resulting in prevention of pregnancy. The FITC labeled SAF was found to bind the entire surface of spermatozoa. Vaginal application and oral administration of rSAF to mice for 14 successive days did not demonstrate any significant change in vaginal cell morphology, organ weight and tissue histology of reproductive and non-reproductive organs and had no negative impact in the dermal and penile irritation tests.ConclusionThe Sperm Agglutinating Factor from Staphylococcus warneri, natural microflora of human cervix, showed extensive potential to be employed as a safe vaginal contraceptive.

Short preheating at 41°C leads to a red blood cells count comparable to that in RET channel of Sysmex analysers in samples showing cold agglutination

AimsThe presence of cold agglutinin in blood samples can cause a spontaneous agglutination of red blood cells (RBCs) when low temperature occurs. This phenomenon causes a spurious lowering of RBC...

High Reproducibility of Adhesion Formation in Rat with Meso-Stitch Approximation of Injured Cecum and Abdominal Wall

Objective: Peritoneal adhesions following surgery are a common, serious pathology with severe complications. Appropriate animal adhesion models are essential for the assessment of adhesion preventing medical devices. This study introduces a variation of an established rat model in which highest degree adhesions are induced with excellent reproducibility (OPAM = optimized peritoneal adhesion model). Thus, this model seems to be eligible to study effects of adhesion preventing devices.Methods: 24 Lewis male rats were divided into four groups (OPAM, WSFX, sham-OPAM, sham-WSFX). The OPAM technique comprised cecal abrasion, creation of an abdominal wall defect plus approximation of injured areas by a suture, which was compared to a setting of lesions without suture fixation (WSFX). All rats were sacrificed at day 7. Macroscopic and histopathological evaluations were performed. Results were statistically analyzed using ANOVA and Dunnett's test.Results: In OPAM rats macroscopic analyses revealed a 90% incidence adhesion of cecum to the abdominal wall, all adhesions imposing as complete agglutination. In WSFX animals incidence of adhesions formation was 75%, while in both sham groups there were no adhesions at all. Histology showed the structure of adhesions with merged smooth muscle of colon and skeletal muscle of abdominal wall in all cases.Conclusion: OPAM technique provides adhesions of injured areas with a better probability than with conventional methods. All OPAM adhesions impressed as highest degree adhesions, i.e. agglutination. Due to high reproducibility in incidence and extend of adhesion formation, the OPAM is recommended for testing of adhesion prevention medical devices.

Labial Adhesions in Pubertal Young Females: A Case Series

Labial adhesions describe a partial or complete agglutination of the labia minora and are most frequently seen in females younger than 6 years of age. There are few previously reported cases of labial adhesions in postpubertal females, most commonly associated with prior genital trauma. We present 6 cases of pubertal females with labial adhesions and no prior history of genital trauma or dermatological vulvar conditions.

Labial agglutination and infection in prepubertal girls

Objectives: To assess the correlation of previous infections with the frequency of labial agglutination, to confirm the effectiveness of oestrogen therapy and the frequency of positive microbiological swabs after treatment. Methods: The study included 88 girls aged from 3 months to 10 years who have been examined at the Gynaecological outpatient unit of Children’s Hospital Zagreb from 2005-2007. Anamnestic data about previous infections were obtained from the parents. All girls were treated with an oestradiol cream twice a day. The patients were then re-examined 2 weeks and/or 4 weeks later. After the treatment, vaginal and vulvar smears were taken for microbiological analysis. Results: Twenty-eight (31.8%) girls were lost to follow up. Out of 60 girls in 27 (45%) girls we learned about a previous respiratory or urinary infection in the 3 months before our exam. 45/60 (60%) girls presented with complete agglutination, but without urine disturbances and 15/60 (25%) had partial fusion. After 2 weeks of treatment we found complete resolution in 16/60 (26.7%) cases, after 4 weeks in 30/60 (50%) cases, and after 4-6 weeks of local hormonal treatment in 14/60 (23.3%) cases. Out of 60 study patients’ culture in 27 (45%) the causative agents were isolated from the vaginal culture. Agglutination subsequently recurred in 20/60 children (33.3%), mostly in association with respiratory infection Conclusion: Topical oestrogen application is the method of choice in the treatment of labial agglutination, but for at least 4 weeks of application. Recurrence of agglutination is common and mostly associated with infection. It could be reduced by application of a lubricant to lysed surfaces for as long as 6 months after lysis and at the time of infection.

N-acetyl-9-O-acetylneuraminic acid, the receptor determinant for influenza C virus, is a differentiation marker on chicken erythrocytes.

Erythrocytes from chicken of different age were analysed for their agglutinability by influenza C virus, which has been shown recently to use N-acetyl-9-O-acetylneuraminic acid as a high-affinity receptor determinant for the attachment to cells. Only with birds not younger than six days complete agglutination of the erythrocytes was observed. The hemagglutination titer which was initially low reached its maximum value at the age of about 20 days. Sialic acid was isolated from erythrocytes, purified and analysed by colorimetry, thin-layer chromatography, high-performance liquid chromatography, and gas-liquid chromatography-mass spectrometry. The sialic acid content of erythrocytes from one-day old and adult chicken was 21 micrograms and 18 micrograms sialic acid/ml packed erythrocytes, respectively. While N-acetylneuraminic acid was the major type of sialic acid on erythrocytes from both one-day old and adult chicken, N-acetyl-9-O-acetylneuraminic acid was only detected on red blood cells from adult animals accounting for 30-40% of total sialic acid. These results indicate that N-acetyl-9-O-acetylneuraminic acid, in addition to serving as a receptor determinant for influenza C virus, represents a developmental marker on chicken erythrocytes.

Surface and somatic antigens of Fusiformis nodosus

Fusiformis nodosus produces at least 2 agglutinating antigens during growth on hoof agar plates. One of these is a surface or K antigen that is removed by washing the bacteria in buffered saline or water. In the presence of homologous antiserum prepared in rabbits or sheep, cells with K antigen exhibit a distinctive agglutination that is characterized by the rapid formation of coarse, flocculent granules, and subsequently by a loosely coherent sediment at the base of the test tubes. The surface antigen is inactivated when F. nodosus is boiled for more than 10 min. F. nodosus grown in biphasic cultures or in hoof broth also retains the surface antigen, but with subcultivation in liquid medium this characteristic is soon lost. Thirty-three isolates of F. nodosus from outbreaks of both benign and virulent foot-rot in sheep were classified into 3 types, A. B and C on the basis of their surface antigen. There was relatively little serological cross-reaction between these 3 types. Another 13 isolates could not be classified by the tests used. Another antigen in the cell wall of F. nodosus is resistant to extensive heating and is responsible for a fine granular agglutination in the presence of antisera. This reaction is best demonstrated with antisera prepared in rabbits and cells that have been washed with isotonic saline. Antisera prepared in rabbits cause complete O-type agglutination of F. nodosus that have been boiled for 1 h., but those prepared in sheep cause partial agglutination of such cells and only at lower dilutions. Tube agglutination and indirect haemagglutination tests with rabbit antisera suggest the presence of an antigen that is common to the 3 F. nodosus serotypes identified by K agglutination tests.

Hemagglutination with adenovirus serotypes belonging to Rosen's subgroups II and 3.

SummaryThe hemagglutination by adenovirus types 1, 2, 4, 5, 6 (subgroup III), and 9, 15 (subgroup II) was studied. Serotype 9 displayed, as should be expected, a complete agglutination of human red cells. In addition these cells were agglutinated partially by all the remaining serotypes, although a considerable variation in agglutinability of human erythrocytes obtained from different donors was demonstrated. The hemagglutination of these red cells by serotype 9 was RDE sensitive, whereas the agglutination of human cells by the remaining serotypes was resistant to this treatment. The partial agglutination of both human O and rat red cells peculiar to unfractionated preparations of members of subgroup III was suggested to be caused by the presence of a relative excess of incomplete HA's (i.e., fibers and pentons), in unfractionated preparations of these types, which competitively interfered with the complete hemagglutinin. The following observations supported this explanation (a) isolated complete HA's, su...

The Effective Points of Steroid Hormone on Rheumatoid Arthritis

The clinical and experimental studies had been performed in order to know the effective points of steroid hormone on rheumatoid arthritis. The methods were as follows:(1) The latex fixation test was examined using the human serum with rheumatoid arthritis, 68 years old, female, stage 3, class 4, classical.(2) The steroid hormone was added on it after the complete agglutination had been observed.(3) The patients' serum was incubated with steroid hormone at 37 C-degree for one hour and then the agglutination test was examined with the latex reagent.(4) The reaction between the 1/2 diluted latex reagent with the physiological salts solution and the serum was demonstrated as a control.(5) The latex test reagent was incubated with steroid hormone and then, the agglutination test was examined with the patients' serum.The results were as follows:In the reaction of (1), (2), (3) and (4), above mentioned, it showed the stronger agglutinating nature. But in the last reaction, in case of using the incubated latex reagent with the steroid hormone it was lesser agglutinated. This facts indicate the effectiveness of steroid hormone on rheumatoid arthritis are not only by the antiinflammatory nature, but also by the direct influences on antigenantibody reaction itself.

Bivalent nature of incomplete anti-D (Rho).

ERYTHROCYTE antibodies were first demonstrated by their ability to agglutinate erythrocytes suspended in saline; these have been termed complete antibodies. In 1944, Wiener1 and Race2 independently showed that certain erythrocyte antibodies were incomplete, possessing the ability to fix to the erythrocyte surface without leading to agglutination. It has been postulated1, and generally accepted, that the complete saline agglutination antibody is bivalent, and the incomplete coating antibody univalent. Recent investigations in our laboratory using antibodies labelled with iodine-131 have led us to question this hypothesis3,4. Our results could best be explained by assuming a bivalent nature of incomplete erythrocyte antibodies.

Agglutinin-adsorption analysis of Vibrio fetus.

Reich, C. V. (Pennsylvania State University, University Park), C. E. Heist, and H. W. Dunne. Agglutinin-adsorption analysis of Vibrio fetus. J. Bacteriol. 82:210-214. 1961.-All pathogenic strains of Vibrio fetus are apparently members of a single serotype. Within this broad grouping, it was possible to define the relationship of certain selected strains by a system of complete reciprocal adsorption and cross agglutination. Formalin treated suspensions of V. fetus, and controlled titer homologous rabbit antisera were used. The results of a set of reactions, involving five strains, were analyzed as a complete reaction pattern. A system of 12 antigens and 4 haptens, each alphabetically characterized, was necessary to reproduce schematically the experimentally obtained reaction pattern. One of the antigens, designated as antigen A, was a somatic antigen present on every strain of V. fetus examined. Evidence indicates that this antigen represented about 5% of the total antigenic area of the cell surface.

Agglutination of bacteria in a three-component system consisting of cells, surface-active cations and electrolyte

Macroscopic agglutination of S. schottmuelleri cells occurs in the presence of appropriate concentrations of cationic surface-active agents and electrolytes. The concentration of cationic agent required to produce complete agglutination in the presence of electrolyte is a function of cell density. The adsorption of surface-active cations prior to agglutination results in the acquisition of hydrophobic properties by the cells. Agglutination by electrolytes can be demonstrated to follow the Schulze-Hardy rule in a manner contrary to prediction: the cations of the electrolytes are the flocculating ion. Cationic surface-active agents of dissimilar structure give a pattern of agglutination which does not correlate with the bactericidal activities of these agents.

Studies on hemagglutination and hemolysis by escherichia coli antisera.

A study on hemagglutination and hemolysis by Escherichia coli O111 and O55 (rabbit) antisera and on hemagglutination and hemolysis inhibition by E. coli O111 and O55 antigens revealed the following facts. 1. Red blood cells of man, dog, rabbit, guinea pig, sheep, rat, and chicken adsorb E. coli O111 and O55 antigens and thus become specifically agglutinable by the homologous E. coli antisera. 2. The adsorption of these E. coli antigens is a function of the concentration of the antigen, the time (from 5 minutes to 2 hours) of treatment of the red blood cells with the antigen, and the concentration of the red blood cells used. 3. Red blood cells of man and sheep adsorb simultaneously both antigens, as indicated by the fact that both antisera give agglutination of all red blood cells. Complete agglutination does not occur when a mixture of red blood cells treated separately with the two antigens is added to one or the other of the two antisera. 4. Treatment of red blood cells of man with one of the antigens does not block the adsorption of the second antigen. Human cells treated with either or both antigens are still agglutinated by the homologous blood group (A, B, and Rh)-specific antibodies. 5. In the presence of guinea pig complement, E. coli O111 and O55 antisera produce hemolysis of modified human red blood cells in titers of the same order of magnitude as those giving hemagglutination and bacterial agglutination. The same antisera produce hemolysis of sheep cells treated with the identical antigens in titers exceeding by far those giving agglutination of modified human or sheep red blood cells. 6. Both sediment and supernate of a boiled E. coli suspension are capable of modifying red blood cells for E. coli hemagglutination; in contrast, the supernate obtained from an unboiled suspension and then heated does not modify red blood cells for hemagglutination, although it contains the antigen which can specifically adsorb E. coli antibodies, as shown by means of the hemagglutination and hemolysis inhibition tests. 7. Both the unheated and the boiled suspensions of E. coli O111 and O55 inhibit hemagglutination and hemolysis specifically. 8. Rabbit red blood cells modified by either E. coli O111 or 055 antigens, upon intravenous injection into rabbits, engender specific E. coli antibodies. The significance of the results is discussed.

The Effect of Virus Quantity and Dilution Procedure Upon the Hemagglutination Inhibition Test for Newcastle Disease Diagnosis

THE agglutination of chicken red blood cells by Newcastle disease virus (NDV) was first observed by Burnet (1942) and Lush (1943). The species having red blood cells susceptible to agglutination by NDV were reported by Clark and Nagler (1943) and Brandly et al. (1946). Studies on the application of the hemagglutination (HA) phenomenon to the diagnosis of Newcastle disease with specific inhibition of agglutination by immune serum resulted in recommendation of procedures for the hemagglutination inhibition (HI) test by Brandley et al. (1947) Beach (1948) and the Bureau of Animal Industry (1946).METHODSThe HI test employed in this study was adapted from the procedures recommended by Brandly et al. (1947) and Beach (1948). For determining the HA titer of NDV, the highest 0.5 ml. 2-fold dilution of virus causing complete agglutination of 0.5 ml. of 0.5 percent chicken red cells was regarded as one HA unit.The regular beta .

Agglutination tests in the diagnosis of enteric fever in the inoculated

1. Early blood culture is the best method of laboratory diagnosis in the inoculated case of enteric fever, but agglutination tests may give useful information.2. In inoculated cases of enteric fever O and Vi agglutinins may be absent or present only at low titres. Steadily rising titres are not the rule and the titres more often fluctuate.3. Complete agglutination at titres over 1: 80 for TO, over 1: 40 for AO, or over 1: 10 for Vi is suggestive but not diagnostic of an active infection.4. Non-specific stimulation of the O and Vi agglutinins in the inoculated affects mainly the lower range of titres which are probably of little diagnostic significance.5. Previous inoculations increase the proportion of people with residual O agglutinins within 6 months of the last inoculation but do not affect the Vi agglutinins.6. Vi agglutinins may be present in normal inoculated persons who are not carriers.

On Infectious Mononucleosis

In the spring of 1941, during the occurrence of an epidemic of infectious mononucleosis, we attempted to isolate a transmissible agent from sterile Berkefeld filtrates of nasal washings and whole blood by inoculation into the chorio-allantoic membrane of the chick embryo. Material was obtained from patients who gave a characteristic history, blood picture, and whose heterophile antibody titers ranged from complete agglutination of 160 to a high of 2560. We also attempted passage from patients whose clinical history and leucocyte picture were typical and who later gave heterophile agglutinations in the range mentioned. The nasal washings and blood from some patients were frozen for 12 hours before they were used. Two additional cases made their appearance this fall. In all, a total of 8 attempts was made to transmit the agent from 8 cases; positive results were obtained from 4. The sterile filtrate, when inoculated onto the membrane, showed small pearl-gray reactions with a spreading cloudiness about them....

The Non-Identity of Jack Bean Agglutinin with Crystalline Urease

Abstract Hotchkiss and Tauber (1) have stated that crystalline urease agglutinates washed rabbit and mouse erythrocytes, but not those of sheep. As little as 0.00014 mgm. of urease is said to have caused a perceptible effect. This work is quite erroneous. We find that washed rabbit and dog erythrocytes are not agglutinated in the slightest when incubated for two hours at 37°C. with as much as 3 mgm. of recrystallized urease. It is true that the jack bean, like many other beans, contains a substance which agglutinates red cells. This is thermolabile and non-dialyzable. We have obtained it in solution in such concentration that 1 cc. will cause an almost complete agglutination of 5 cc. of 2 per cent erythrocyte suspension in five minutes at room temperature. Diluted 100 times the material will produce an agglutination detectable by microscope within thirty minutes at 37°C. This 100-fold diluted solution contained only about 0.00015 mgm. of urease per cubic centimeter.

Acid Agglutination Zones of Typical and Variant Pneumococci.

The present basis for differentiation between typical and variant forms of pneumococci involves a subjective opinion of colony morphology. It became desirable, in the course of our studies, to develop a method for this differentiation depending entirely on some objective physical measurement. Such a method has been found to be available by utilization of the phenomena of agglutination of organisms in solutions of varying acidity. This method has been of service in recognition of variants of other bacterial species., Gillespie has studied the acid agglutination zones of several types of typical pneumococci and found them to be similar. The organisms to be tested were grown in pneumococcus broth, the period of incubation being kept as short as consistent with adequate growth, usually about 8 hours. The organisms were packed by centrifuging and washed 3 times with saline. After the final centrifugation, they were restored to original volume with saline. 0.5 cc. quantities of this suspension were added to a similar volume of standard buffer solution of the desired acidity, incubated for 3 hours at 56°, and then recorded. The buffer system used was that of Northrup and deKruif. Results obtained with various strains are shown in the accompanying table. These results indicate that the zones of acid agglutination for variant strains of pneumococci, either occurring spontaneously or derived through laboratory manipulation, are widely separated from those of typical strains. The strains possessing the wide zone of complete agglutination possess also the characteristics accompanying the “rough” colony form.

Brucella Agglutinins in the Blood and Milk of Cows

In connection with the routine agglutination test of blood samples from dairy and beef cattle herds for bovine infectious abortion (Brucella infection) an opportunity has been provided on different occasions to note the correlation of specific agglutinins for Brucella species in the blood and milk of cows. Over a period of years the presence of Brucella agglutinins in the blood serum and their absence in the milk serum * of the same animal has been observed. Notwithstanding that these observations have repeatedly shown that agglutinins in the milk are not reliable guides in the diagnosis of Brucella infection in cattle, few comparisons to show the relation of agglutinins in the blood and milk serums of the same animals have been reported. The comparative results of the blood and milk serum agglutination reactions in the same animals included in this report are based upon the standard tube or slow agglutination method, using two dilutions?1: 50 and 1: 200. Complete agglutination in both dilutions or complete agglutination in 1: 200 was regarded as positive. A positive 1: 50 and a negative 1:200 was regarded as doubtful. The blood samples were collected from the jugular vein and the milk samples from the four quarters of the udder. The clear blood serum for the agglutination test was obtained in the usual way by centrifugation, or by allowing the blood samples to stand until the clear serum escaped from the clot. The milk serum, or whey, was obtained by adding one drop of rennin to 4 or 5 cc. of milk and, after incubating for 30 minutes at 37 C, the clot was broken and the clear whey brought to the surface by centrifugation. In our experience the relative proportion of negative milk serum agglutination tests in lactating animals that react positively to the blood test are illustrated in the following results obtained from two groups of dairy cows. In herd 1 of 91 animals, 77, or 84.62^, of the blood and milk samples gave complete agreement in the test for Brucella agglutinins. Fifty-two,