Biomedical subjects
Publications and source records attributed to J de Graaff.
Malignant struma ovarii is a very rare tumor, belonging to the group of germ cell tumors. The diagnosis is difficult and there is no agreement in the literature concerning the necessary histologic features for malignancy. A case report is presented and the problems of diagnosing are discussed.
Killing of Proteus mirabilis by human polymorphonuclear leukocytes was tested in the presence of different Bacteroides species. In vitro experiments showed that anaerobic bacteria interfered with the killing of aerobic bacteria. However, this inhibitory effect was not a property of all Bacteroides species. Bacteroides gingivalis W83 showed the greatest inhibitory effect of the five Bacteroides strains tested. Killing of P. mirabilis was inhibited by the culture supernatant of B. gingivalis but not by washed cells. Two factors were found in the supernatant of B. gingivalis to account for the inhibitory effect. One was heat stable with a molecular weight of less than 3,500 and inhibited the killing activity of leukocytes, and the other was heat labile and partly inactivated the complement system. The killing experiments paralleled chemiluminescence measurements.
The role of hemolysin in the nephropathogenicity of Escherichia coli was studied in a hematogenous pyelonephritis model in mice. The nephropathogenicity of a nonhemolytic, avirulent E. coli strain was increased by simultaneous injection with its hemolytic, nephropathogenic parent. This helper mechanism could be attributed to hemolysin, since the simultaneous injection of partially purified hemolysin gave a similar enhancement of nephropathogenicity. Intraperitoneal injection of hemoglobin or iron sulfate before intravenous challenge with this avirulent strain also led to increased virulence. The nephropathogenicity-enhancing effect of hemolysin is therefore supposed to depend on increasing the level of available iron in the host. Under conditions of plentiful iron, hemolysin production was repressed, as shown by in vitro growth experiments in the presence of exogenous iron. These results suggest that the production of hemolysin is regulated by feedback inhibition.
Thirty saccharolytic and asaccharolytic black-pigmented Bacteroides strains were tested for their susceptibility to vancomycin. All asaccharolytic strains appeared to be partly or completely inhibited at a concentration of 7.5 micrograms/ml, whereas most saccharolytic strains were resistant to this concentration. The use of vancomycin in Bacteroides selective media is discussed.
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The prevalence of beta-haemolysis and yellow pigmentation among 89 isolates of Streptococcus mutans from human dental plaque and 59 Strep. mutans isolates from positive cultures of human dental root canals were investigated. beta-Haemolytic strains were more frequent among the root-canal isolates (25.4 per cent) than among the dental plaque isolates (12.4 per cent). The distribution of pigment production among beta-haemolytic strains (22 pigmented strains out of 26 strains) and among non-haemolytic strains (5 pigmented strains out of 122 strains) showed a relationship between haemolysin and pigment production. beta-Haemolytic, pigmented strains (biotype c and e and 35-39 per cent guanine plus cytosine) were common in plaque and were the dominant type of Strep. mutans in some plaques. As beta-haemolysis by Strep. mutans was only demonstrated after strict anaerobic growth, whereas pigment production was demonstrated after growth under less anaerobic conditions (candle jar) on sucrose-containing agar plates, the observation of pigment may facilitate a presumptive identification of beta-haemolytic strains of Strep. mutans.
The virulence of faecal and urinary Escherichia coli strains was studied in relation to serotype, haemolysin production and haemagglutination pattern. By means of an experimental mouse model E. coli strains can be divided into avirulent (I), mouse nephropathogenic (II), and generally virulent (III) strains. Virulent group II and group III strains were more often haemolytic and haemagglutinating than avirulent group I strains. Presence of K antigen could not be associated with virulence. Discriminant analysis for qualitative variables revealed that no combination of the investigated properties contributed more to a strain's virulence level than did one single property. It is concluded that other virulence factors, apart from haemolysin production in group II strains and haemagglutinins in group III strains, must be involved in the determination of a strain's virulence level. All O2, O6 and O18 ac strains tested were virulent, and by far the most O75 strains were avirulent, whereas other O groups were more variable with regard to virulence. Pyelonephritis strains were more often mannose-resistance haemagglutinating than faecal and other urinary isolates, indicating that mannose-resistant adhesins may be important in the pathogenesis of pyelonephritis.
Culture filtrates of all eight strains of Bacteroides gingivalis and all five strains of B. asaccharolyticus were toxic for Vero cells. Cytotoxicity was in general greater with material from cultures of B. gingivalis than from B. asaccharolyticus but none of the culture filtrates from eight strains of B. melaninogenicus showed activity in this test. The toxic material was released during prolonged incubation and more detailed study of preparations from one strain indicated that it had a molecular weight of less than 3500 and was heat stable.
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The nephropathogenic Escherichia coli strain P673 was shown to harbor two plasmids with molecular sizes of 70 and 41 megadaltons, respectively. The 70-megadalton plasmid, pCW1, coded for tetracycline resistance, whereas hemolysin production was coded by the 41-megadalton plasmid, pCW2. Plasmid pCW1 proved to be self-transmissible, in contrast to pCW2. Transfer of the hemolysin character was associated with the appearance of a 110-megadalton plasmid, pCW3. The incompatibility of pCW3 with both native plasmids and restriction enzyme analysis led to the conclusion that pCW3 is a cointegrate of pCW1 and pCW2, pCW2, carrying the hemolytic determinant, is involved in the nephropathogenic character of strain P673, because (i) elimination of pCW2 from P673 was associated with a loss of virulence and (ii) the nephropathogenicity of the avirulent mutant could be restored by reintroduction of pCW2 DNA as part of a cointegrate structure.
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The virulence of B. gingivalis strain W83 was studied in an experimental animal model. Cells grown overnight, washed and resuspended in broth, were injected intradermally or subcutaneously in the back of guinea pigs, rats and mice. This strain proved to be very virulent, causing a severe phlegmonous abscess in guinea pigs. Also in mice, which are thought to be resistant to infections with black-pigmented Bacteroides strains, the same type of infection could be induced. Rats proved to be rather insensitive. The model presented can be used as a simple virulence test for these anaerobic bacteria.
E. coli strains were isolated from the urine of patients with acute cystitis in general practice and from the faeces of a comparable reference group of healthy individuals. These strains were serotyped and tested for virulence in an experimental mouse model. Of 30 cystitis-strains 18 were virulent, and of 30 faeces-strains 15 were virulent. It is concluded that the cystitis-strains were not more often virulent than the faeces-strains. O antigens commonly found among urinary E. coli isolates were present in 60% of the cystitis-strains and in 37% of the faeces-strains. K antigens commonly found in urinary E. coli strains were present in 33% of the cystitis-strains and in 12% of the faeces-strains. Neither the presence of common urinary O-antigens, nor the presence of common urinary K antigens could be associated with virulence of the isolated strains. However, it is suggested that certain O and K antigens (O2,O6, K23) may be associated with virulence for the urinary tract.
The influence of haemolysin production on virulence was studied in an experimental mouse model. Urinary strains of Escherichia coli can be divided into three virulence groups by determining their kinetics in the mouse kidney after intravenous injection. Virulent strains of groups II and III were more often haemolytic than avirulent group-I strains. Haemolytic virulent strains often caused haemoglobinuria in the mice, and killed the mice more rapidly than did non-haemolytic virulent strains. No relationship was found between alpha-haemolytic activity and virulence in wild-type haemolytic strains. When haemolysin production was reduced or eliminated by treatment with actinomycin-D or rifampicin, six out of seven group-II strains tested gave the same results as avirulent group-I strains. However, the kinetics in the mouse kidney of four haemolytic group-III strains tested was not changed after reduction or elimination of haemolysin production; only a small decrease in toxicity was observed. It is concluded that haemolysin production by E. coli is a decisive virulence factor in most of the mouse-nephropathogenic group-II strains, but not in the virulent group-III strains.
Beta-hemolytic strains of Streptococcus mutans were identified and physiological reactions and hemolysin production were studied. Strict anaerobic conditions were shown to be indispensable for the expression of beta-hemolysis on blood agar plates; hemolysis was only observed after growth in an atmosphere with an oxygen content of 10 ppm or lower. The occurrence of beta-hemolytic strains was found not to be restricted to one biotype of S. mutans. Two hemolytic activities were detected in liquid cultures. Oxygen-labile activity was detected in prereduced broth cultures and was shown to be unstable and indetectable in supernatants. Solubilization of oxygen-labile hemolysin was achieved by the addition of Tween 80 to the cultures. Oxygen-stable activity was detected in supernatants of cell suspensions in the presence of maltose, Casamino Acids, and Tween 80. In contrast to oxygen-labile activity, oxygen-stable activity was not detected in broth cultures.