Knowledge and educational needs of individuals with the factor V Leiden mutation.

BACKGROUND: Genetic testing for factor (F)V Leiden is widely performed in an effort to prevent thrombosis-related morbidity. The implications of a positive test for patients' health perception and the extent of patients' understanding of results are not known. OBJECTIVES: This study examined patient experience of genetic testing for FV Leiden. PATIENTS AND METHODS: The study was a cross-sectional, mailed survey of 110 patients who tested positive for the FV Leiden gene mutation at an academic medical center between 1995 and 2001. Patient knowledge about FV Leiden, satisfaction with available information, and psychosocial reactions to testing were assessed and the influence of demographic and clinical characteristics on outcome measured. RESULTS: The magnitude of thrombosis risk associated with FV Leiden was incorrectly estimated by 79% of participants. Many patients (64%) stated that they had not been given much information about FV Leiden and 68% still had many questions. Most patients (53%) felt that their healthcare providers do not understand FV Leiden. Patients who had been seen by a hematologist or in a specialized thrombosis clinic were more knowledgeable and had less information need. Most patients (88%) were glad to know genetic test results, despite negative psychosocial implications such as increased worry (43%). CONCLUSIONS: Knowledge of genetic status increases awareness of thrombosis risk among patients, but magnitude of risk is often overestimated. Affected individuals indicate that there is a lack of available information about FV Leiden and that additional educational resources are needed.

[The identification of Bacillus cereus, Bacillus lichenformis and Bacillus subtilis strains using the coagglutination reaction]. / Versuch zur Erfassung von Bacillus-cereus-, Bacillus-licheniformis- und Bacillus-subtilis-Stämmen mittels der Co-Agglutinationsreaktion.

Polyclonal rabbit antisera against 4 B. cereus strains were selected from a total of 9 B. cereus antisera and pooled. This serum agglutinated all available B. cereus strains (n = 63) at a titre > or = 1:64 when tested by a slide co-agglutination reaction. One hundred and thirty-six Bacillus strains belonging to 17 other species reacted with higher serum concentrations only (titres mainly < or = 1:16). A pooled antiserum comprised of two B. licheniformis antisera and two B. subtilis antisera agglutinated all 43 B. licheniformis strains and all 38 B. subtilis strains at a titre > or = 1:128, but one strain each of the species B. pumilus, B. macerans and B. fastidiosus was also agglutinated at a titre 1:128.

Isolation and partial characterization of the major protein of the outer membrane of Pasteurella haemolytica and Pasteurella multocida.

The N-terminal amino acid sequence of the 35 kDa (p35) major outer membrane protein (MOMP) of P. multocida shared a strong homology with those of homotrimeric nonspecific porins of gram-negative bacteria. The capacity of outer membrane protein (OMP) preparations of P. multocida to bind to respiratory mucosal surface preparations was inhibited significantly by using a polyclonal anti-p35 antiserum in an adhesion ELISA. Anti-p35 antiserum cross-reacted with a 44 kDa (p44) MOMP of P. haemolytica. N-terminal sequencing of MOMP p44 revealed a homology of 81% with the putative porin MOMP p35 of P. multocida.

[In vitro adherence of Pasteurella haemolytica to tracheal mucins and a tracheal epithelial cell preparation from cattle]. / In vitro Adhärenz von Pasteurella haemolytica an Trachealmucin und einer Trachealepithelzellpräparation vom Rind.

Outer membrane protein preparations (OMP) of 11 Pasteurella haemolytica field isolates of the serovars A1, A2, A11 and non-typable strains from cattle were extracted by N-lauryl-sarcosine sodium salt. Capsular extracts were prepared by heat treatment. Both preparations and whole cell suspensions bound to a preparation of an epithelial cell wall fraction of trachea and to tracheal mucus of cattle. Binding was demonstrated by enzyme linked immunosorbent assay (ELISA). Distinct high binding values were shown by the OMP and the capsular extracts of the serovar A1-strains.

In vitro binding of Campylobacter jejuni/coli outer membrane preparations to INT 407 cell membranes.

Outer membrane fractions (OMs) of nine Campylobacter (C.) jejuni and two C. coli strains belonging to different serovars, from human and various animal origins, were extracted by treatment with sodium N-lauryl sarcosinate. Using n-octyl-beta-D-glucopyranoside a 42-kDa protein and a flagella-enriched fraction were obtained. The capacity of the crude bacterial OM preparations, the purified 42-kDa protein and the flagella to bind to membranes of the human embryonic intestinal cell line INT 407 was tested by an enzyme-linked immunosorbent assay. The crude OM and the 42-kDa-enriched fraction were found to bind very well to the cell membranes, whereas the flagella preparation showed only a weak binding. Using monoclonal antibodies (mAbs) with HS 2-lipopolysaccharide (LPS) specificity, binding of crude HS 2 strain OM preparations to cell membranes was detected in a significant range, whereas with flagellin-specific mAbs binding of OMs and flagella to cell membranes was only detected to a very low extent. Binding of OMs to cell membranes was inhibited by preincubation of OMs with serovar-specific mouse hyperimmune serum, whereas on preincubation with mAbs directed against LPS or flagella binding was practically not inhibited. OMs extracted after pretreatment of the bacteria with proteinase K showed an altered SDS-PAGE pattern especially for the 42-kDa protein subunit and and their capacity to bind to cell membranes was significantly reduced. The binding was also reduced by preincubation of the OMs with L-fucose or D-mannose.

In vitro binding of Pasteurella multocida cell wall preparations to tracheal mucus of cattle and swine and to a tracheal epithel cell wall preparation of cattle.

Outer membrane preparations of various Pasteurella isolates (Pasteurella multocida and some other Pasteurella species) from cattle and swine were extracted by N-lauryl-sarcosine sodium salt. Capsular extracts were prepared by heat treatment. Both preparations bound to epithel cell wall preparations (ECW) of trachea from cattle and to tracheal mucus of cattle and swine. Binding was demonstrated by an enzyme-linked immunosorbent assay (ELISA). Distinct high adherence values were shown by the greater part of membrane preparations of mucoid Pasteurella strains, especially when originating from cattle.

[Activation of the resident microbial flora as a possibility for the improvement of humoral antibody potentials]. / Aktivierung der residenten Keimflora als Möglichkeit zur Verbesserung des humoralen Antikörperpotentials.

The effect of a parenteral application of an adjuvant (Propionibacterium acnes) and an adjuvant/antigen combination (Staphylococcus aureus Cowan I/Al(OH)3, Pseudomonas aeruginosa/Al(OH)3 respectively, was tested with regard to the improvement of antibacterial resistance. Parameter was the humoral antibody production (IgG) of rabbits against six facultative pathogen bacterial species (St. aureus, Sc. faecium, Bac. cereus, P. multocida, E. coli, Ps. aeruginosa) and against sheep rod blood cell membranes. The three methods of treatment led to a significant enhancement of IgG-antibodies against the particular homologous antigen. In addition to this specific reaction the application of adjuvant/antigen combinations provoked a significant enhancement of antibodies also against heterologous antigens (P. multocida, Bac. cereus, sheep red blood cell membranes). The application of P. acnes had no distinct effect on the antibody titer against the different antigen preparations. In order to analyse the immune response qualitatively, a greater part of serum samples was examined by immuno blot technique. The number of partial antigens recognized by the sera increased during the experiments but the rise did generally not relate to developments of antibody titers. Nevertheless, the Ps. aeruginosa/Al(OH)3-treatment seemed to improve the ability of sera to detect antigenic determinants of heterologous bacterial species.

Crossed immunoelectrophoresis applied to representative strains from 11 different Pasteurella species under taxonomic aspects.

Crossed immunoelectrophoresis evaluated on a numerical basis revealed a close antigenic relationship between species of the genus Pasteurella. By cluster analysis, 4 groups on similarity levels between 87% and 72% S could be separated which were connected by a minimum level of 69.5% S. One subgroup included all biovars or subspecies consisting of strains with mucoid growth. Another feature governing the antigenic relationship seemed to be the host range of Pasteurella species. Despite a considerable number of cross-reacting antigens, representative strains of the genera Haemophilus and Actinobacillus were clearly separated from Pasteurella. Similarly, "Pasteurella" haemolytica and Taxon 16 strains tested did not belong to this genus. An Escherichia coli strain showed a higher number of cross-reacting antigens, confirming known antigenic relationship among Gram-negative bacterial species.

[Electron microscopic studies of mucosal extirpates of rabbits after after in vitro contact with Pasteurella multocida subsp. multocida strains]. / Elektronenmikroskopische Untersuchungen an Schleimhautexstirpaten von Kaninchen nach in vitro-Kontakt mit Pasteurella multocida subsp. multocida-Stämmen.

In vitro experiments of adhesion and colonisation of mucosal surface fragments (nose, trachea) from freshly killed rabbits with four strains of Pasteurella multocida subsp. multocida with different types of capsule antigen (A, D, without capsule) showed the following results: Above all the capsule serovar A strain, producing a capsule of hyaluronic acid and fimbria, were able to adhere and to form microcolonies on the mucosal surface. Microvilli of epithelial cells and mucus producing cells were recognized as the place of adhesion. The formation of microcolonies occurred with a destruction of the kinocilia, which was caused by a bacteria free culture filtrate as well.

In vitro binding of Campylobacter jejuni surface proteins to murine small intestinal cell membranes.

Surface proteins of nine Campylobacter jejuni strains belonging to three different serovia were extracted with lysozyme/ethylenediamine-tetraacetic acid. The preparations bound to isolated murine small intestinal cells and to a membrane fraction (MF) of isolated brush borders obtained by detergent treatment with Triton X-100 and Nonidet P40. Binding was demonstrated by an enzyme-linked immunosorbent assay procedure. Using lipopolysaccharide (LPS)- and flagella-specific antisera the contribution of flagella and LPS, present in the protein preparations, to the total binding to MF was investigated. Only up to approximately 10% of the total binding of each strain was found to be mediated by LPS, 10%-33% of binding was flagella dependent. The preparations of four strains (serovar HS2) bound in a trypsin-sensitive manner (45%-85% reduction), while the others (serovaria HS1 and HS13) were hardly influenced by trypsin treatment. Binding to MF was not impaired by preincubation of the bacterial surface protein preparations with several sugars and lectins.

Species identification and serotyping (capsular antigen) of Pasteurella strains from sheep flocks in south Germany and in Syria.

135 Pasteurella strains were cultivated from nasal swabs of sheep as well as pneumonic lungs of dead and slaughtered sheep. The specimen originated from 41 flocks in South Germany and from 15 flocks and 60 slaughter sheep in Syria (Hama region). Serovariety A2 prevailed amongst P. haemolytica strains (6) isolated in South Germany (53 strains) and in Syria (41 strains). In addition 10 further serovarieties were identified in South Germany (next frequent were A8, A1 and A6) and 7 in Syria. Untypable strains appeared to be more frequent in Syria. Other Pasteurellae (17) represented 1/4 of isolates in Syria and 1/3 of isolates in South Germany. Species identification resulted in P. multocida ssp. multocida (25), P. multocida ssp. septica (4 strains, Syria only), P. canis (3 strains, South Germany only) and Pasteurella-like strains (9 strains). Twelve P. multocida ssp. multocida strains carried capsular antigen D and 7 capsular antigen A. In most cases where multiple samples were examined from one flock, strains with different capsular antigens and/or belonging to different Pasteurella species were isolated (max. 8).

Examination of 20 Bacillus species by crossed immunoelectrophoresis under taxonomic aspects.

In order to examine the efficiency of crossed immunoelectrophoresis (CIE) for the differentiation of species and for the determination of taxonomic relationships within the genus Bacillus, an investigation including strains of 20 species was performed. Ultrasonic extracts (USE) of sporefree-grown vegetative cells were used. With rabbit antisera against USE of type strains, the CIE was accomplished in homologous and heterologous combinations. USE's of additional strains were included into this investigation. In order to evaluate the immunoelectropherograms, the number of precipitates was counted. A mean of 103 precipitates was found in homologous reactions. With few exceptions, heterologous combinations showed less similarity in the number of antigens. Because of their high cross-reactions, the following species could not be differentiated: B. subtilis from B. amyloliquefaciens and strains of B. coagulans, furthermore, B. cereus from B. thuringiensis. The different species revealed lower numbers of precipitates with decreasing taxonomic relationships in heterologous combinations. This observation was used to classify the investigated species by a 'position-frequency analysis' (PFA). After sorting the matrix of precipitate numbers in the ascertained optimal sequence of species, a cluster analysis was carried out. The phenogram showed 6 (respectively 8) group clusters. The members of the morphologic group I (Smith et al., 1952) was found only in group cluster 2. The phenogram was partly in agreement with phenograms based on other characteristics, e.g. DNA hybridization.

Represent the batches 2 of strain NCTC 3991 and NCTC 3992 B. coagulans?

In a previous publication (M. Hartung and E. Hellmann: Examination of 20 Bacillus Species by Crossed Immunoelectrophoresis under Taxonomic Aspects, Zbl. Bakt. Hyg. A 263 (1987) 509-524) we came to the conclusion that the species B. subtilis and B. coagulans were closely related. This statement has to be revised: The antigenic relationship is low. Further investigations have shown that all three B. coagulans-strains, although originating from different sources, were overgrown by B. subtilis. The same fact may have caused errors in other publications (e.g. Parry et al.: A Colour Atlas of Bacillus Species, Wolfe Medical Publications Ltd., London, 1983.

[Growth rates of two virulence plasmids carrying Yersinia enterocolitica after contamination of heated milk, raw minced pork and vegetables]. / Vermehrungsstudien an zwei virulenzplasmidtragenden Yersinia enterocolitica-Stämmen nach Kontamination von erhitzter Milch, rohem Schweinehackfleisch und Vegetabilien.

The psychrotrophic organism Y. enterocolitica (Y.e.) is able to grow in foodstuffs thus rendering it dangerous for the consumer. Among others, the preparation of food in the kitchen might be an important way of transmission. Not only contaminated raw meat but aswell latently infected humans and pet animals (cats, dogs) may serve as a reservoir for Y.e. contamination. After artificial contamination of UHT milk and vanilla sauce with subsequent storage at 4 degrees C, two Y.e. strains of serovars 0:3 and 0:9 carrying the virulence plasmid multiplied during the first 6 d (9 d) at a rate of 1 log/d (Fig. 1). In pasteurized milk the multiplication rate was 0,65 log/d (Fig.2). Maximum bacterial counts in UHT milk after 15 d were in the order of 10(9) CFU/ml (Fig.4). In pasteurized milk, even minimal inoculation doses of less than 5 CFU/100ml (calculated) produced multiplication of Y.e. to 1,7-27 X 10(4) CFU/ml after 7 d (Tab.1). In this case the milk has been contaminated by an appropriately diluted suspension of faeces from infected mice. When selected colonies were tested on MOX-Agar, a loss of the virulence plasmid could not be observed. In minced fresh pork meat with a bacterial count of 4,2 X 10(6) CFU/g--most of them i.e. 2,6 X 10(6) belonging to Pseudomonas sp.--added Y.e. did not multiply, but viable organisms could be recovered after contamination with only 2,9-6,6 X 10(2) CFU Y.e./g and storage at 4 degrees C for 72 h (Tab.2). Similarly, after contamination of quartered paprika husks and halves of carrots, Y.e. strains could be re-isolated after 15 d storage at 4 degrees C or room temperature if the number of bacteria used for inoculation was in the range of 10(6) (Tab.3). Feeding contaminated carrots to NMRI-mice a calculated number of 5 X 10(3) CFU Y.e./mouse were sufficient to cause infection of part of the animals together with shedding of the organisms through the faeces (Tab.4). These results support the assumption that milk and home made milk products like vanilla sauce create a risk for human health when they are stored for a few days after contamination with Y.e. carrying the virulence plasmid. The same may be true for raw minced pork and fresh vegetables but only if the initial contamination was high. This might occur during food processing procedures.(ABSTRACT TRUNCATED AT 400 WORDS)