Inside Poland.
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Biomedical subjects
Publications and source records attributed to M Pollack.
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Serum antibodies to Pseudomonas aeruginosa exotoxin A and immunotype-specific lipopolysaccharides were evaluated as diagnostic and prognostic markers in patients with Pseudomonas disease. Hemagglutination titers to exotoxin A were 1:1,024 or higher and/or showed a fourfold acute-to-convalescent increase in 17 of 25 (68 percent) patients infected with Pseudomonas compared with only one of seven (15 percent) colonized (p = 0.01) and two of 24 (8 percent) culture-negative patients (p less than 0.001). By comparison, hemagglutination titers to the lipopolysaccharide of patients' Pseudomonas isolates were 1:1,024 or higher or showed a fourfold increase in only four of 17 (24 percent) infected patients and in none of six (0 percent) colonized patients (p = 0.96). Serial antibody titers to exotoxin A provided serologic confirmation of invasive disease, distinguished infection from colonization, and, in the case of decreasing titers, indicated successful therapy. It is concluded that serum antibodies to exotoxin A are useful serologic markers for the clinical assessment of Pseudomonas infections in man.
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Twenty-seven lots of human immune globulin for intravenous use (IGIV) from seven different producers, including one hyperimmune preparation, were examined for immunologic reactivity and opsonic and protective activity against Pseudomonas aeruginosa. All IGIVs contained hemagglutinating antibodies to seven immunotype-specific lipopolysaccharides of P. aeruginosa (geometric mean titer +/- SE, 14 +/- 3 for nonhyperimmune preparations and 420 for the hyperimmune product) and to exotoxin A (77 +/- 15). All IGIVs tested demonstrated opsonic activity against P. aeruginosa in an in vitro granulocyte-dependent bactericidal assay. All IGIVs conferred dose-dependent protection (hyperimmune more so than nonhyperimmune) against fatal burn-wound infections due to P. aeruginosa in mice. In contrast, single lots of hyperimmune and nonhyperimmune IGIV conferred limited protection against infections due to P. aeruginosa in granulocytopenic mice. These studies indicate the potential prophylactic efficacy of IGIV in human pseudomonas disease, and the possible need for high doses of hyperimmune IGIV in granulocytopenic patients.
We studied the relationship between serum antibodies to the cross-reactive endotoxin core of Escherichia coli and survival following Pseudomonas aeruginosa septicemia. Core glycolipid was purified from the outer cell membrane of a uridine diphosphate galactose 4-epimerase-deficient rough mutant E. coli (J5 strain), characterized, and used as the antigen in a quantitative enzyme-linked immunosorbent assay (ELISA) to measure core-specific IgG and IgM antibodies. 43 patients with Pseudomonas septicemia, among whom there was a mortality of 42%, were evaluated. Core-specific antibody concentrations in acute sera ranged from 1 to 49 micrograms/ml in the case of IgG and from 1 to 200 micrograms/ml for IgM. Core-specific antibodies of both isotypes were higher in patients who survived compared with those who succumbed to their septicemias (mean, microgram/ml +/- SEM, 26 +/- 3 vs. 14 +/- 4, P = 0.005 for IgG, and 55 +/- 12 vs. 18 +/- 5, P = 0.009 for IgM). Although total IgG levels were also higher in acute sera from survivors compared with nonsurvivors (mean, mg/dl +/- SEM, 1,120 +/- 99 vs. 694 +/- 119, P = 0.004), total IgM levels were virtually identical in the two groups (146 +/- 23 vs. 148 +/- 48, P = 0.52). Conversely, patients with core-specific IgG levels greater than 10 micrograms/ml at the onset of septicemia had better survival than those with levels less than 10 micrograms/ml (79 vs. 14%, P less than 0.001), and patients with core-specific IgM levels greater than 30 micrograms/ml had better survival than those with levels less than 30 micrograms/ml (81 vs. 44%, P = 0.01). In comparison, patients with total IgG levels greater than 1,000 mg/dl also had better survival than those with levels less than 1,000 mg/dl (82 vs. 42%, P = 0.01), while those with total IgM levels greater than 150 mg/dl showed somewhat less improvement in survival compared with those with levels less than 150 mg/dl (71 vs. 50%, P = 0.12). Core-specific IgM was highly correlated with core-specific IgG (r = 0.52), but not with type-specific anti-lipopolysaccharide (r = 0.13) or anti-toxin A (r = 0.12) antibodies, or with total IgG (r = 0.28) or IgM (r = 0.31). In contrast, core-specific IgG correlated somewhat more closely with type-specific antibodies (r = 0.36), and with total IgG (r = 0.51) and IgM (r = 0.52). Stepwise linear discriminant analysis indicated that type-specific antibody levels were the best predictor of outcome, among those antibodies examined, followed by anti-core IgM. Although anti-core IgG, anti-toxin A, and total IgG levels all correlated individually with survival, none augmented the prognostic power of type-specific antibodies in combination with anti-core IgM, which together predicted outcome accurately 73.5% of the time. Host factors not significantly associated with anti-core antibody levels included rapidly fatal underlying disease, age, sex, leukopenia, and prior treatment with cytotoxic drugs. In contrast, prior steroid therapy was associated with low levels of both core-specific IgG and IgM (P < 0.05). These data suggest cross-protective activity against P. aeruginosa septicemia of naturally occurring antibodies to the endotoxin core of E. coli. Anti-core antibodies, particularly of the IgM isotype appear to augment the more specific protective immunity engendered by antibodies to the O-specific side chains of Pseudomonas lipopolysaccharides. This cross-protective immunity likely applies to other Gram-negative pathogens as well.
A prolonged partial thromboplastin time in a patient was investigated, as well as those of his family members. One sibling, the mother, and the patient all shared a lupus-like anticoagulant and low Factor XII levels in association with the A 11, Bw 35, DR 4 haplotype previously found in familial lupus erythematosis. The rest of the family had normal coagulation profiles and the entire family had normal findings when tested for ANA, complement and immune complexes. After being confirmed in a second coagulation laboratory, the lupus-like anticoagulants in the three affected members of the family disappeared upon retesting 3-8 weeks later.
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Toxoid was prepared by treating exotoxin A of Pseudomonas aeruginosa with formalin. Coincubation of exotoxin A for two to four weeks at 37 degrees C with 0.5% formalin and 10(-3) M lysine followed by one to two weeks of storage in the absence of these reagents reduced cytotoxicity, preserved antigenicity, and minimized subsequent reversion of the toxoid. Formalin enhanced the adenosine diphosphate (ADP)-ribosyl transferase activity of the toxin while decreasing its toxicity, whereas formalin plus lysine reduced both. Although no antigenic changes were detected by immunodiffusion analysis, analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed minor alterations in the toxin structure. Toxoid induced high titers of antibody to exotoxin A in the sera of mice and rabbits. Antiserum to toxoid neutralized mouse lethality, cytotoxicity, and ADP-ribosyl transferase activity of untreated exotoxin A. Toxoid-immunized mice were resistant to large doses of exotoxin A administered iv.
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Pseudomonas aeruginosa exotoxin A, a potent inhibitor of eukaryotic protein synthesis, is produced in vivo during human infection. We tested the hypothesis that exotoxin A may be responsible for the leukopenia which sometimes accompanies pseudomonas disease by examining the in vitro toxicity of exotoxin A for human bone marrow granulocyte-macrophage progenitor cells (colony-forming units in culture [CFU-c] in the soft agar cloning system. Colony formation by freshly obtained marrow cells from five normal subjects was inhibited by exotoxin A in a concentration-dependent manner. The mean 50 and 100% inhibitory concentrations of toxin were 1.4 x 10(-10) and 1.4 x 10(-8) M, respectively, and significant inhibition was observed at a toxin concentration as low as 1.4 x 10(-13) M in two subjects. The inhibitory effect of exotoxin A on colony formation was specifically neutralized by antiserum to exotoxin A. Although mouse CFU-c were somewhat less sensitive to exotoxin A in vitro compared with human CFU-c, exotoxin A produced significant leukopenia in vivo in mice. These data suggest a possible mechanism for the leukopenia which sometimes occurs in human pseudomonas disease.
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Cryptic 21-hydroxylase deficiency has been previously described in asymptomatic family members of patients with classical congenital adrenal hyperplasia (CAH). These family members were detected by high baseline 17-hydroxyprogesterone levels found in the course of family studies. The hormonal responses to ACTH of the family members with cryptic 21-hydroxylase deficiency were determined and compared to the responses of patients with CAH, patients with acquired adrenal hyperplasia, family members predicted to be heterozygous for CAH, family members predicted to be unaffected, and the general population. The ACTH-stimulated levels of 17-hydroxyprogesterone and delta 4-androstenedione in the cryptic family members were elevated above the level of the general population or family members heterozygous for classical CAH, but below that of patients with CAH. The hormonal profile of patients with cryptic 21-hydroxylase deficiency is similar to that of patients with acquired adrenal hyperplasia. The response of family members heterozygous for the cryptic gene (21-OH CRYPTIC/21-OH NORMAL) was indistinguishable from that of family members heterozygous for the classical CAH gene (21-OH CAH/21-OH NORMAL). These studies support our previous proposal that patients with cryptic 21-hydroxylase deficiency are genetic compounds, having one gene for a severe enzyme deficiency and one gene for a mild 21-hydroxylase deficiency. Thus, the 21-hydroxylase genotype in cryptic 21-hydroxylase deficiency is 21-OH CAH/21-OH CRYPTIC.
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Endocarditis is a rare manifestation of disseminated Histoplasma capsulatum infection. A 22-year-old man presented with a seven month history of fever, weight loss, and progressive aortic insufficiency. The diagnosis of H. capsulatum was suggested by a diagnostic rise in complement fixation titers and positive echocardiographic findings. The diagnosis was confirmed prior to surgery by positive bone marrow culture. Progressive congestive heart failure necessitated replacement of the aortic valve which subsequently grew H. capsulatum. In this case, a combination of amphotericin B therapy and valve replacement was curative.
During a study of efficacy of amantadine prophylaxis of influenza A virus infection in young adults, gross and subtle side effects were monitored. Eighty-eight students were randomly selected to receive either amantadine or placebo for four weeks or both in a sequential crossover design of two weeks each. Side effects (i.e., dizziness, nervousness, and insomnia) occurred in 33% of those receiving amantadine and in 10% of those receiving placebo (P less than 0.005). Although side effects were well tolerated by most subjects, six volunteers discontinued amantadine because of marked complaints. Cessation of side effects occurred in more than half of those continuing amantadine. Sixteen students receiving amantadine had decreased performance on sustained attention tasks as compared with ones receiving placebo (P less than 0.05). Gross and subtle side effects of amantadine observed in this study on currently recommended dosage are higher than previously reported, which may be an important factor in consideration of mass prophylaxis.
Recurrent respiratory infections associated with "mucoid" Pseudomonas aeruginosa characterize the advanced stages of cystic fibrosis. To determine if chronic antigenic stimulation is associated with circulating immune complexes (CIC), we assayed the sera of 20 hospitalized patients using the technique of precipitation with 4% polyethylene glycol. Elevated CIC levels, defined by > 310 micrograms IgG per ml, were found in 18 of 20 patients, (range, 350 to 3200 micrograms/ml). Serum, supernatant, and resuspended precipitates were assayed for hemagglutinating antibodies against pseudomonas lipopolysaccharide (LPS or endotoxin) and exotoxin A antigens. Both serum anti-LPS (range, 1:64 to 1:2048) and antitoxin titers (range, 1:64 to 1:16, 384) were markedly elevated and higher than titers in supernatants and resuspended precipitates, indicating antibody excess. "Enrichment" ratios for antibodies present in CIC were calculated by proportion of titer to immunoglobulin in the precipitated complex relative to these values in serum. Mean enrichment ratios of 13.1 and 13.9 were obtained for LPS antibody before and after 2 mercaptoethanol reduction, but the mean enrichment ratio for antitoxin was only 2.07. Serially diluted supernatants and precipitates were boiled for 1 hr and tested for endotoxin-like activity by the limulus test. At > 1:8 dilutions, precipitates were positive, and supernatants were negative. These findings indicate that CIC's are common in advanced cystic fibrosis, and analysis of the precipitated complexes demonstrates significant (> 13-fold) enrichment of antibodies against LPS but not exotoxin antigens, as well as endotoxin-like activity in boiled precipitates.