The relationships between antitoxic titers determined in a group of young people before and after booster ADT by the in vivo neutralization test. Comparative NT-PHAR evaluations

The geometrical mean of antitoxic titers determined in a lot of immunized young men who received, 5-7 years before, a booster with diphtheria-tetanus bivaccine, was of 1.54 (x/divided by 10.54) I.U./ml. At one month after another booster, performed with Tetanus toxoid (adsorbed), an increase of 34 times was recorded, the geometrical mean reaching the value of 52.67 (x/divided by 10.42 I.U./ml). The covering coefficients of the minimal protective antitoxin level have increased from 154 to 5267, in the estimation performed at the level of geometrical mean (calculating the ratio between the value of this indicator and that of the protection limit) and from 1623 to 54935, or from 15 to 505, respectively, in the estimations performed at the levels of the limits of the statistical range of one geometric standard deviation (Mg x/divided by D Sg). The study of the relations between pre- and post-antitoxic titers, performed by the linear regression analysis, leads to an equation whose slope evidenced the lower amplitude of the booster titers, along the increase of previous titers. The study of the inter-methods (neutralization-passive haemagglutination) regression allowed to obtain a correlation coefficient, between methods, of r = 0.83 and to perform the transposition of the minimal protective N.T. level (0.01 I.U.) in the terms of passive haemagglutination reaction, at the titer of 160 (0.3 PHAU/ml), for the p less than or equal to 0.05 probability level.     

Usefulness of the passive hemagglutination micromethod in the diagnosis of whooping cough

Studies on modification of diagnostic test for pertussis have been continued, they were practically restricted to an application of passive haemagglutination micromethod. Six hundred and twenty eight sera from children suspected to be infected with Bordetella and 38 sera of control children suffering from non-infectious diseases were tested. Despite of the fact that higher titers were obtained with method using red blood cells preserved in Alsevier solution, the passive haemagglutination micromethod may be used in field studies especially during pertussis epidemics what was confirmed by statistical analysis.     

GAST after 3 years' use]

Increasing of systematic syphilis screening has led many scientists to think to automatisation of classical cardiolipid reactions. The Debains-Kolmer reactions have been adapted to continuous flow but they appear actually inefficient. The use of ART (automated reagin test) gives very satisfying results and this reagent has obtained its homologation in the United States. The apparition of Groupamatic has incitated M. GARRETTA to adapt on this material the K. Antigen reagent manufactured by the Blood Transfusion Center in Lille and has led to the definition of GAST reagent (Groupamatic automated syphilis test). The authors describe the method of preparation of this reagent and its utilization on Groupamatic. It appears that the bovin cardiolipidic extract, non specific by definition, lays down to antagonist problems: to be sensitive enough when specific enough. It's the final reagent settlement which is the more delicate and which needs the maximum in manipulation. Yet, the composition of GAST is now well established and this reagent, ready for use, may be kept during three months at 4 degrees C. Then the authors describe the results of their own use of GAST, in routine on Groupamatic 360: out of 6 079 plasmas of blood donnors examined, 3 per cent of reactions are positive or doubtfull (which stays compatible with the screenings in large number). After confirmation with complementary tests made with manually GAST, RPR with microscopic reading, haemagglutination, and lastly fluorescent method, it appears that the rate of positive reactions is 1,7%. This result is conformable to habitual statistics. In conclusion, the GAST is a well-adapted reagent for Groupamatic technology, and represents a progress compared with classical manually methods. The adaptation of syphilis screening on Groupamatic is a factor of rentability of this equipment, allowing to realize 360 tests in one hour. At last, considering that cardiolipidic reactions are not of an absolute diagnostic value, treponemic complementary tests are necessary in order to confirm positive results picked up on Groupamatic.     

A study of factors affecting the sensitivity of the passive haemagglutination method for serotyping Campylobacter jejuni and Campylobacter coli and recommendations for a more rapid procedure

Factors affecting the sensitivity of the passive haemagglutination method for serotyping campylobacters have been studied. The concentration of red blood cells during the haemagglutination stage of the procedure markedly affected the titer obtained. An increase in concentration of red blood cells resulted in a lower titer, with titers being inversely proportional to red blood cell concentration. No differences in titer were observed when erythrocytes were sensitized at a range of pH values between pH 5.0 and pH 8.0. The time required for antigen extraction and for red blood cell sensitization was shown to be 15 min each, thus resulting in a reduction in the time required for serotyping. Furthermore, use of avian erythrocytes enabled the haemagglutination reactions to be read after incubation for only 1 h. Combining these procedures with a rapid slide haemagglutination test enables a single worker to serotype over 100 C. jejuni and C. coli isolates within 1 working day.     

Saccharides of seven Pseudomonas aeruginosa immunotypes

Abstract The structures of O-specific polysaccharides obtained by mild acid degredation of lipopolysaccharides (LPS) from seven Pseudomonas aeruginosa Fisher's immunotypes have been studied. The polysaccharides consist mainly of monoamino and diamino sugars, frequently also carrying acidic functions. Some of the sugars were detected in nature for the O-specific polysaccharides of the immunotypes 2, 3, 4, 5 and 6 are identical to those of the polysaccharides of the 011; 0(2a)2c; 01; 010a, 10b and 07a, 7d Lányi-Bergan serological subgroups respectively, whereas no analogues have been found for the immunotypes 1 and 7. Some cross-reactions between the LPS of different immunotypes were observed in passive haemagglutination tests; the results of inhibition of passive haemagglutination and agar gel immunoprecipitation point, however, to a specificity of the LPS. Many of the LPS of the seven Pseudomonas aeruginosa immunotypes manifest rather a high cross-protective activity in active immunization tests in mice. The nature of the cross-protective activity of the LPS is discussed.     

Problems posed by the preparation of specific anti-cytomegalovirus immunoglobulins

The authors describe the preparation of a first batch of intravenous cytomegalovirus (CMV) immune globulin at the Nancy Regional blood transfusion centre. Immune plasmas were selected from 3 640 healthy volunteer blood donors on the basis of CF antibody titers to CMV (Kolmer's method modified) of, at least, 1:8; plasmas from approximately 10% of the donors were therefore selected. The 68 liters of pooled immune plasma had à CF antibody titer of 1:16 (CMV antibody titers of 1: 10 000 and 1: 640 when tested in the ELISA assay and passive hemagglutination assay respectively). Intravenous immune globulin was produced from pooled plasma by Cohn fractionation and treatment with pepsin at pH 4; 4.8 liters of immune globulin were prepared and divided in 96 doses of 50 ml each. The final product was found to have a CMV antibody titer of 1: 32 (CF) 1: 50 000 (ELISA) or 1: 2 560 (passive hemagglutination). Recent reports on the preparation of CMV immune globulin are briefly reviewed.     

Saccharides of seven Pseudomonas aeruginosa immunotypes

The structures of O-specific polysaccharides obtained by mild acid degradation of lipopolysaccharides (LPS) from seven Pseudomonas aeruginosa Fisher's immunotypes have been studied. The polysaccharides consist mainly of monoamino and diamino sugars, frequently also carrying acidic functions. Some of the sugars were detected in nature for the first time in these organisms. The structures of the O-specific polysaccharides of the immunotypes 2, 3, 4, 5 and 6 are identical to those of the polysaccharides of the 011; 0(2a)2c; 01; 010a, 10b and 07a, 7d Lányi-Bergan serological subgroups respectively, whereas no analogues have been found for the immunotypes 1 and 7. Some cross-reactions between the LPS of different immunotypes were observed in passive haemagglutination tests; the results of inhibition of passive haemagglutination and agar gel immunoprecipitation point, however, to a specificity of the LPS. Many of the LPS of the seven Pseudomonas aeruginosa immunotypes manifest rather a high cross-protective activity in active immunization tests in mice. The nature of the cross-protective activity of the LPS is discussed.     

Changes in collagen metabolism--another look at osteoarthrosis

Collagen types I and III were isolated from osteoarthrotic cartilage. Immunofluorescence study has shown that these two collagen types are present in the deep layers of cartilage. Additional staining for fibronectin revealed the presence of the cell-adhesive protein in osteoarthrotic cartilage. Neither collagen type I and type III nor fibronectin were found in control cartilage. A passive haemagglutination assay determined anticollagen antibodies (against types I, II and III) in sera of some osteoarthrotic patients.     

Glomerulonephritis Associated with Antibody to Glomerular Basement Membrane

Glomerulonephritis associated with antibody to glomerular basement membrane, shown by linear staining of the glomerular basement membrane with fluoresceinated anti-IgG antisera, was found in only 10 out of 400 (2·5%) renal biopsy specimens studied by immunofluorescence. Seven of these cases had rapidly progressive glomerulonephritis, five with lung haemorrhage (Goodpasture''s syndrome) and two without, and three had less severe nephritis without lung haemorrhage. Circulating antibody to glomerular basement membrane, measured by a passive haemagglutination technique and by indirect immunofluorescence, was detected in the serum of all patients with rapidly progressive glomerulonephritis by both techniques but only by the passive haemagglutination method in two of the other three patients. Two patients died of their lung haemorrhage, one despite bilateral nephrectomy, and lung haemorrhage and circulating antibody to glomerular basement membrane persisted after bilateral nephrectomy in another patient.     

Rapid and simultaneous screening and quantitation of antitetanus and anti-HBs antibodies using Laurell's method]

Every transfusion Center which must hunt out anti-tetanus and anti-HBs antibodies to obtain specific corresponding immunoglobulins, is submitted to rapidity, precision and low cost necessity. In order to satisfy these obligations, the authors describe a simple and rapid method, allowing use of the same serum, through a single stage, and simultaneously, screening and quantitation of anti-tetanus antibodies (according to Laurells' method) and of fractionation suitable anti-HBs antibodies (through electro immunodiffusion). Technical conditions exposed in this paper allow direct quantitation of anti-tetanus antibodies from 1 to 30-40 IU; dilutions can be made for upper titers. False negative reactions due to zone effect, as observed in EID and passive haemagglutination do not occur while using this method. Its routine use contributes to the increase of the percentage of fractionation suitable plasmas, without overloading the screening laboratory's tash.     

Detection and titration of antitetanus antibodies using an automated passive hemagglutination method]

An automated reversed passive haemagglutination technique is described, using human erythrocytes sensitized by the chromic chloride method with higly purified tetanus toxoid. Such erythrocytes are agglutinated by the specific antibodies in a Technicon autoanalyzer. Clots are decanted, supernatant is hemolyzed and optical density is measured at 550 millimicron. Assay standards for comparison, ranging from 5 to 25 UI/ml, are prepared from human antitetanus immunoglobulins titrated by toxin neutralisation assay in mice. This method allows to screen donors' sera for presence of high titers of tetanus antibodies (larger than or equal to 5 UI) suitable for preparation of antitetanus immunoglobulins. 4,4% of donors have circulating antitetanus antibody levels corresponding to 5 UI/ml or more, by this method. The specificity of antibodies has been confirmed by the neutralisation assay in mice. The results obtained among 1.000 donors well agree with those of the counter-immuno-electrophoresis technique (C.E.P.). But, when, compared with C.E.P., the haemagglutination assay appears more objective, more quantitative and sensitive, and allows to get not only a rapid screening test but also a precise titration simultaneously.     

Comparative methods of detecting HbsAg in chronic glomerulonephritis patients in Nigeria

Abstract A comparative evaluation of the enzyme-linked immunosorbent assay (Elisa), passive haemagglutination (PHA) and counterimmunoelectrophoresis (CIE) methods were carried out. Approx. 6% of control samples were positive with the Elisa assay, while the values for Behring Institute (BI) passive haemagglutination, Wellcome Research Laboratory (WRL) passive haemagglutination and CIE were 2.7%, 2.2% and 3.3%, respectively. The above data contrast with values of 21.3%, 10.6, 10.3 and 12.3 obtained in sera from chronic glomerulonephritis patients.
Our observation suggests that HbsAg may be associated with chronic glomerulonephritis.     

Immunochemical studies on sialic acid-containing lipopolysaccharides from enterobacterial species

Abstract Lipopolysaccharides with sialic acid as a component were isolated from six bacterial serovars of the family Enterobacteriaceae . SDS-polyacrylamide gel electrophoresis, immunoblotting and passive haemagglutination demonstrated that epitopes are localized in the O-specific polysaccharide of lipopolysaccharides. The strains were tested for a serological relationship; the only distinct cross-reactivity recorded was between Escherichia coli serovars O24 and O56. The molecular mimicry phenomenon was found for Citrobacter freundii serovar O37 which shared epitopes with horse as well as human red blood cells.     

Characterization of an outer membrane protein associated with haemagglutination and adhesive properties of enteroaggregative Escherichia coli O111:H12

Diarrhoeagenic Escherichia coli strains of serotype O111:H12 are characterized by their aggregative pattern of adherence on cultured epithelial cells and thus are considered enteroaggregative E. coli (EAEC). We have previously shown that these EAEC strains lack the genes encoding the aggregative fimbriae I and II described in other heterologous EAEC strains. In this paper, we show compelling data suggesting that a plasmid-encoded outer membrane 58 kDa protein termed aggregative protein 58 (Ap58) produced by EAEC O111:H12 strains, is associated with the adherence capabilities and haemagglutination of animal red blood cells. This conclusion is supported by several lines of evidence: (i) adherent O111:H12 strains are able to produce Ap58; (ii) non-adherent O111:H12 strains are unable to produce Ap58; (iii) antibodies raised against Ap58 inhibited adherence and haemagglutination of epithelial and bovine red blood cells, respectively; (iv) a non-adherent E. coli K-12 host strain containing the ap58 gene determinant on plasmid pVM15 displayed abundant adherence to cultured HEp-2 cells; and (v) the purified Ap58 bound specifically to HEp-2 and bovine red blood cells. Our findings indicate that the aggregative adherence in the O111:H12 strains may be also mediated by non-fimbrial adhesins. We believe our data contribute to the understanding of the adherence mechanisms of these organisms.     

Adhesion ofHelicobacter pylori strains toα-2,3-linked sialic acids

Helicobacter pylori is a human pathogen associated with gastritis and peptic ulcer. Adhesion properties ofH. pylori to various structures have been described in the literature, including evidence for sialic acid-binding. To study the specificity and frequency of sialic acid-binding, fourteenH. pylori strains were investigated using haemagglutination with derivatized erythrocytes carrying sialic acids only on defined glycans and using haemagglutination inhibition assays. From these studiesH. pylori strains can be grouped into sialic acid-dependent and sialic acid-independent classes. The sialic acid-dependent strains require -2,3-linked sialic acid for haemagglutination. The potential roles of sialic acid-dependent adhesions forH. pylori-related infections are discussed.Abbreviations Sia sialic acids - Neu5Ac N-acetyl-neuraminic acid - Neu5Gc N-glycolylneuraminic acid - Neu5Fm N-formylneuraminic acid - Neu5TFA N-trifluoroacetylneuraminic acid - RBC human red blood cells (erythrocytes)     

Immunochemical and biological studies of antigens isolated from a strain of Bacteroides fragilis

Phenol/water-extracted lipopolysaccharide and a fraction HM, extracted with acetate buffer pH 2.0, from Bacteroides fragilis strain 62/73 are antigenically different as shown by immunodiffusion, passive haemagglutination, haemagglutination inhibition and preliminary chemical investigations. Biological activity, assessed with the local Shwartzmann reaction, was demonstrated for the lipopolysaccharide whereas antigen HM was almost inactive in this test. HM is immunogenic in rabbits. Antibodies against HM were detected in seven out of ten sera of healthy humans.     

Rough mutants of Salmonella typhimurium: immunochemical and structural analysis of lipopolysaccharides from rfaH mutants

Lipopolysaccharides, extracted by phenol/chloroform/petroleum ether, from two rough mutants of Salmonella typhimurium of class rfaH were studied by passive haemagglutination inhibition and by methylation analysis. The structural and immunochemical analyses showed that (i) formation of the galactose I unit of the core is defective, but the defect is not complete, and (ii) of those core chains which do receive the galactose I residue, many are not continued to form complete core, but instead terminate at intermediate points. This suggests that the rfaH gene, though involved in formation of the galactose I unit, is not the structural gene for the galactosyltransferase which adds this unit. The rfaH product may be a positive regulator for several rfa genes specifying glycosyltransferases, or it may be a protein needed for the efficient action of several such transferases.     

Determination of seven unconjugated steroids in the blood and seminal plasma of the fertile male rabbit.

Seven unconjugated steroids were measured in the blood and seminal plasmas of fertile male rabbits by radioimmunoassay. The blood plasma testosterone concentration was 4--5 times that of the seminal plasma. Dehydroepiandrosterone, estrone and 17beta-estradiol were found in measurable amounts in the blood plasma; however, these steroid levels were slightly lower in seminal plasma. Androstenedione and 5alpha-dihydrotestosterone were present in equal quantities in both the seminal and blood plasmas. By contrast, seminal plasma pregnenolone level was about twice that of the blood plasma. The determination of seminal plasma steroids may lend itself as a complementary assessment to blood steroid determinations for the evaluation of the normal function of various reproductive organs.     

Quantification of plasma-derived blood coagulation factor VIII by real-time biosensor measurements

Plasma-derived blood coagulation factor VIII was analyzed in real time using biosensor technology. Monoclonal antibodies directed against the heavy and against the light chain of factor VIII were immobilized on different carboxymethyl dextran surfaces. Different factor VIII concentrations were injected over the antibody surfaces in parallel and response levels were determined from the dissociation phase at a fixed time after sample injection. Serial dilutions of plasma-derived factor VIII with known concentrations determined by a commercial FVIIIC:Ag ELISA were used as standards. A quantification limit of 0.9 I.U./ml with antibody 530p and 1.5 I.U./ml with antibody 531p was calculated. Intra-assay precision expressed as percent coefficient of variation was below 10% for concentrations above 0.6 I.U./ml. Inter-assay precision for antibody 530p was below 20% for concentrations higher than 0.6 I.U./ml. For 531p, inter-assay precision was below 10% for concentrations higher than 2 I.U./ml. A sensor chip lifetime in respect to regeneration of at least 100 cycles for both antibodies was found. The small sample requirement of 35 μl allows fast analysis of different FVIII products and the use of two monoclonal antibodies directed against two different FVIII domains provides additional information about the integrity of the FVIII molecule.     

Further experience in rehabilitation of zone II flexor tendon repair with dynamic traction splinting

A review of all flexor tendon repairs in the "no man's land" performed from January of 1985 to June of 1987 was done to evaluate the efficacy of our method of rehabilitation. There were 60 fingers (57 patients) with complete laceration of the flexor digitorum profundus and flexor digitorum superficialis tendons in zone II. Fingers with phalangeal fractures, joint injuries, or significant skin loss were excluded. Follow-up ranged from 12 to 48 months. Rehabilitation consisted of a 12-week protocol using the U.S. military combined regimen of controlled motion. Features from the technique of controlled active extension against rubber band passive flexion as well as those of controlled passive extension and passive flexion were incorporated. The palmar pulley modification of Kleinert's dynamic traction splint was utilized. Strickland's total active motion formula was employed to determine results. The results were classified into the four categories of excellent, good, fair, and poor. Fifty-two fingers (86 percent) were rated excellent, 4 fingers (7 percent) were rated good, 1 finger (2 percent) was rated fair, and 3 fingers (5 percent) were rated poor.