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J S Knapp

Publications and source records attributed to J S Knapp.

At least 19 recordsLinked to original sources

Correlation between in vitro antimicrobial susceptibilities and beta-lactamase plasmid contents of isolates of Haemophilus ducreyi from the United States.

We determined the susceptibilities of 94 strains of Haemophilus ducreyi isolated in various municipalities in the United States between 1982 and 1989 to the following antimicrobial agents: amoxicillin-clavulanic acid, ceftriaxone, erythromycin, azithromycin, ciprofloxacin, ofloxacin, trimethoprim, and spectinomycin. Ceftriaxone (MIC, less than or equal to 0.008 micrograms/ml), azithromycin (MIC, less than or equal to 0.125 micrograms/ml), erythromycin (MIC, less than or equal to 0.125 micrograms/ml), ciprofloxacin (MIC, less than or equal to 0.25 micrograms/ml), and ofloxacin (MIC, less than or equal to 0.25 micrograms/ml) were highly active against all isolates. Amoxicillin-clavulanic acid (MICs, 0.25 to 8.0 micrograms/ml), trimethoprim (MICs, 0.06 to 16.0 micrograms/ml), and spectinomycin (MICs, 2.0 to greater than or equal to 32.0 micrograms/ml) were less active against these isolates. Isolates possessing the 5.7-MDa beta-lactamase plasmid were less susceptible to erythromycin, trimethoprim, and spectinomycin than were isolates possessing the 3.2-MDa beta-lactamase plasmid. The susceptibilities of plasmidless isolates to erythromycin, trimethoprim, and spectinomycin were distributed bimodally; the median MIC for the more susceptible plasmidless isolates corresponded to that for isolates with the 3.2-MDa plasmid, and the median MIC for the less susceptible plasmidless isolates corresponded to that for isolates with the 5.7-MDa plasmid. Thus, plasmid profiles may be valuable markers for geographical variations in antimicrobial susceptibilities of H. ducreyi strains that may indicate the relative efficacy of regimens for the treatment of chancroid. Of the regimens recommended by the U.S. Public Health Service for the treatment of chancroid, our results support the use of erythromycin, ceftriaxone, and ciprofloxacin, and perhaps ofloxacin, but suggest that amoxicillin-clavulanic acid and sulfamethoxazole-trimethoprim should be used with caution.

Anti-Bacterial Agents

Prevalence of gene sequences coding for hypervariable regions of Opa (protein II) in Neisseria gonorrhoeae.

Opas (protein IIs) are a family of surface-exposed proteins of Neisseria gonorrhoeae. Each strain of N. gonorrhoeae has multiple (10-11) genes encoding for Opas. Identifiable elements in opa genes include the coding repeat within the signal sequence, conserve 5' and 3' regions, and hypervariable regions (HV1 and HV2) located within the structural gene. N. gonorrhoeae strains appear to have many biological properties in common that are either HV-region-mediated or associated with the presence of specific HV regions, suggesting that HV regions could be found in many clinical isolates. Oligonucleotides from three source strains representing three conserved regions of opa, 12 HV1 regions, and 14 HV2 regions were used by dot blot analysis to probe 120 clinical isolates of N. gonorrhoeae. The probe for the coding repeat hybridized to all 120 strains, the 3' conserved-region probe reacted with 98% of the strains, and the 5' conserved-region probe with 90% of the strains. Nine HV1 probes hybridized to 3.3-39.2% of the strains, and 13 of the HV2 probes hybridized to 1.7-25% of the isolates. Analysis of the number of probes that hybridized to each of the isolates showed that 19% did not hybridize with any of the HV1 probes and 25% did not hybridize with any of the HV2 probes. Approximately three-quarters of the isolates hybridized with one, two or three of the HV1 probes or one, two or three of the HV2 probes; 89% of the isolates hybridized to least one HV1 or one HV2 probe. The data indicate that some genes encoding HV regions of N. gonorrhoeae Opa proteins are widely distributed in nature.

Bacterial Outer Membrane Proteins

Binding of S protein by Neisseria gonorrhoeae and potential role in invasion.

An agglutination assay was used to examine the binding of purified human S protein (vitronectin, serum spreading factor) to 201 clinical isolates of Neisseria gonorrhoeae. Strains belonging to the protein IA serovars were significantly (P less than 0.001) more reactive in agglutination tests with human S protein and were more serum resistant than strains belonging to the protein IB serovars. The strains from patients with disseminated infections belonged predominantly to the IA serovar (19 of 23) and, with the exception of IA-4 and certain IB serovars, avidly agglutinated with S protein. The serovar IA-4 and IB strains isolated from joint or cerebrospinal fluid failed to agglutinate with S protein and appeared to be less serum resistant than most other IA isolates. Cysteine hydrochloride or 2-mercaptoethanol inhibited agglutination of S protein and a more than twofold increase in resistance to killing by fresh human serum following preincubation with S protein; the serum-sensitive parent strain did not agglutinate S protein, and serum resistance was not increased following preincubation with this protein. Binding of S protein by gonococci may represent a novel pathogenic mechanism that can contribute to serum resistance.

Agglutination

Novel plasmid combinations in Haemophilus ducreyi isolates from Thailand.

Thirty isolates of Haemophilus ducreyi collected in Thailand in 1984 were characterized by plasmid content. Three novel plasmids with estimated molecular masses of 1.8, 2.6, and 2.8 MDa were observed in 29 isolates, in addition to the 3.2-, 5.7-, and 7.0-MDa beta-lactamase and 4.4-MDa sulfonamide resistance plasmids. At least three of the seven plasmids were observed in each of the 29 isolates. The number and diversity of plasmids observed in these isolates of H. ducreyi distinguish them from strains previously described.

Chancroid

Molecular characterization of Haemophilus ducreyi by ribosomal DNA fingerprinting.

Intraspecies genotypic heterogeneity among Haemophilus ducreyi isolates was examined by using genomic fingerprints with rRNA from Escherichia coli as a probe. DNA from 44 isolates of H. ducreyi was digested by restriction endonuclease HincII or HindIII, separated by agarose gel electrophoresis, transferred to nylon membranes, and hybridized with 32P-labeled 16S and 23S rRNA. HincII digests yielded four hybridization patterns (ribotypes), whereas HindIII digests yielded eight ribotypes. Four HincII and five HindIII ribotypes were observed among 14 H. ducreyi isolates collected within a period of 1 month in Kenya, where chancroid is endemic. In contrast, one HincII and two HindIII ribotypes were observed among 28 isolates collected during the Orange County, Calif., chancroid epidemic that occurred in 1981 and 1982. The plasmid content, in conjunction with ribotyping, provided additional differentiation among some isolates of H. ducreyi. This study demonstrates that ribotyping of H. ducreyi may be used to study the epidemiology of chancroid.

Bacterial Typing Techniques

National surveillance of antimicrobial resistance in Neisseria gonorrhoeae. The Gonococcal Isolate Surveillance Project.

The Gonococcal Isolate Surveillance Project is a national sentinel surveillance system to estimate levels and monitor trends of antimicrobial resistance in prospectively collected isolates of Neisseria gonorrhoeae. Of 6204 isolates evaluated from 21 clinic sites between September 1987 and December 1988, 21% met at least one of the surveillance criteria for resistance to penicillin, tetracycline, cefoxitin, or spectinomycin; 2.2% were penicillinase-producing N gonorrhoeae; 1.0% had high-level plasmid-mediated tetracycline resistance; and 16.8% of the isolates without plasmid-mediated resistance had chromosomally mediated resistance (defined as a minimum inhibitory concentration greater than or equal to 2 micrograms/mL) to penicillin, tetracycline, or cefoxitin. Three isolates were resistant to spectinomycin. All isolates were susceptible to ceftriaxone. Resistant isolates were identified from all participating centers. Patient demographic and behavioral characteristics were not predictive of infections caused by resistant organisms. These results demonstrate the wide distribution of antimicrobial-resistant N gonorrhoeae and support recent changes in Centers for Disease Control therapy recommendations for gonococcal infections that no longer recommend tetracycline and penicillin as first-line therapies.

Adolescent

Current status of serotyping of Neisseria gonorrhoeae.

Protein I has become the basis of serotyping of N. gonorrhoeae. A panel of 12 anti-protein I monoclonal antibodies is supplied by Syva Company, Palo Alto, CA, through distributors for research purposes only. These distributors are responsible for storing the monoclonal antibodies, supplying research groups and training new users, particularly in the interpretation of coagglutination reactions. A European Workshop was held in Heidelberg, Federal Republic of Germany in July 1988 to discuss serotyping with these antibodies. The current uses of serotyping of N. gonorrhoeae include epidemiological studies, clinical purposes and surveillance of antibiotic resistance and plasmid carriage. Predominant serovars may be subtyped either by additional antibodies or with the use of another technique, such as auxotyping, determination of antibiotic sensitivities, plasmid analysis or genetic fingerprinting. Conversely, there is growing evidence that it may be appropriate to group certain serovars together. Data collected during prevalence studies could be combined with clinical information and an international data bank set up. This supposes a close future collaboration of all groups involved in the epidemiology of N. gonorrhoeae.

Antibodies, Monoclonal

Standardization of disk diffusion and agar dilution susceptibility tests for Neisseria gonorrhoeae: interpretive criteria and quality control guidelines for ceftriaxone, penicillin, spectinomycin, and tetracycline.

A six-laboratory study developed a standardized method for determining the susceptibilities of Neisseria gonorrhoeae strains to penicillin, tetracycline, spectinomycin, and ceftriaxone. Three quality control organisms were also selected, and quality assurance guidelines were initially generated for the disk diffusion and agar dilution methods. The medium recommended for gonococcal susceptibility testing was GC agar with a defined "XV-like" supplement. The supplement should be free of cysteine, a component implicated in the inactivation of some newer beta-lactam compounds. Penicillin, tetracycline, spectinomycin, and ceftriaxone were stable in agar plates stored at 3 to 5 degrees C for at least 2 weeks. Numerous GC agar and drug disk lots were used during the trials without significant variation in test results. Several other gonococcal strains were recommended for additional medium quality assurance. The disk quality control zone limits were established for N. gonorrhoeae ATCC 49226 (formerly CDC F-18) and Staphylococcus aureus ATCC 25923. MIC quality control ranges were also developed for N. gonorrhoeae ATCC 49226 and S. aureus ATCC 29213. The interpretive criteria for penicillin were as follows: susceptibility, greater than or equal to 47 mm (diameter of inhibition zone) (less than or equal to 0.06 micrograms/ml [MIC]); resistance, less than or equal to 26 mm (greater than or equal to 2 micrograms/ml). For tetracycline they were as follows: susceptibility, greater than or equal to 38 mm (less than or equal to 0.25 microgram/ml); resistance, less than or equal to 30 mm (greater than or equal to 2 micrograms/ml). For spectinomycin they were as follows: susceptibility, >/= 18 mm (</= 32 micrograms/ml); resistance, </= 14 mm (>/= 128 micrograms/ml). For ceftriaxone susceptibility, the criterion was >/= 35 mm (</= 0.25 micrograms/ml) with no other category. Criteria of </= 19 mm for plasmid-mediated resistances to penicillin and tetracycline were also confirmed. The false-susceptibility errors were </= 1% with these criteria.

Ceftriaxone

Comparison of isolates of Neisseria gonorrhoeae causing meningitis and report of gonococcal meningitis in a patient with C8 deficiency.

We studied a previously healthy 20-year-old woman who presented with gonococcal meningitis. The gonococcal isolate, HT-1, was prototrophic by auxotyping, was protein I serovar IB-1, and agglutinated with wheat germ lectin. This isolate differed from the proline-requiring, serovar IA-1 and IB-4, wheat germ-agglutination-negative gonococcal isolates recovered from three patients during a recent outbreak of gonococcal meningitis in Philadelphia. HT-1 was killed by normal pooled human sera (greater than or equal to 98% at 30 min) but not effectively killed by the convalescent-phase sera of the patient (greater than 30% survival at 30 min). Similar results were obtained when mucosal and cerebrospinal fluid isolates from a Philadelphia patient were exposed to these sera, but mucosal and blood isolates from another Philadelphia case showed increased resistance to killing by normal pooled human sera. Further characterization revealed multiple differences in outer membrane and cellular proteins and lipopolysaccharide between case isolates. Absence of the L8 lipopolysaccharide epitope was noted for all isolates. Sera of our patient were found to have low total hemolytic complement (CH100 = 21 U/ml; normal = 55 to 100 U/ml) due to deficiency of C8 (C8 less than 1,000 CH50 U/ml; normal = greater than or equal to 16,000 CH50 U/ml). This is the first reported case of gonococcal meningitis occurring in a patient with a terminal-complement deficiency. Gonococcal meningitis is a rare complication of gonococcal bacteremia. Both defects in host defenses (e.g., terminal-complement deficiency) and organisms with unusual virulence appear to contribute to the pathogenesis of this complication of gonococcal bacteremia.

Adult

Gonorrhea.

Laboratory methods for the isolation and identification of Neisseria gonorrhoeae are updated. The antimicrobial susceptibility patterns of the gonococcus are changing and becoming less predictable. Methods for monitoring susceptibility are presented. The use of serotyping, auxotyping, and molecular techniques to characterize gonococcal isolates and the epidemiological applications are reviewed.

Gonorrhea

Presence of antibodies to the major anaerobically induced gonococcal outer membrane protein in sera from patients with gonococcal infections.

Anaerobically grown Neisseria gonorrhoeae induces and represses the synthesis of outer membrane proteins. One of the anaerobically induced proteins, Pan 1, reacted strongly on Western blots with sera from patients with uncomplicated gonococcal infection, pelvic inflammatory disease, and disseminated gonococcal infection, but not with normal human serum. The pattern of reactivity of the sera against Pan 1 from several gonococcal strains suggested that the protein was antigenically heterogeneous, containing both common and unique epitopes. Staphylococcus aureus V8 protease digestion of Pan 1 from four gonococcal strains revealed the presence of common peptides, with one strain also containing some unique peptides and lacking others. The class of the antibody reactive with gonococcal outer membrane antigens was examined; anti-Pan 1 antibody was found to be IgG or IgM, but not IgA. The IgM antibody present reacted predominantly with Pan 1. These data indicate that the Pan 1 protein is expressed in vivo and strongly suggest that N. gonorrhoeae can grow anaerobically in vivo.

Anaerobiosis

Randomized comparative study of 0.5 and 1 g of cefodizime (HR 221) versus 1 g of cefotaxime for acute uncomplicated urogenital gonorrhea.

Uncomplicated urogenital and concomitant oropharyngeal gonorrhea in 424 male and female patients was treated in a randomized comparative study with 0.5 g of cefodizime (89 men and 54 women), 1 g of cefodizime (87 men and 52 women), or 1 g of cefotaxime (86 men and 56 women). The cure rates were 100% for men and women in the group given 0.5 g of cefodizime, 100% for men and women in the group given 1 g of cefodizime, and 99% for men and 100% for women in the group given 1 g of cefotaxime. The MICs of cefodizime and cefotaxime for the isolate of Neisseria gonorrhoeae ranged from 0.004 to 0.06 micrograms/ml. Chlamydia trachomatis was isolated before treatment in 15% and after treatment in 13% of all patients. Side effects, such as nausea, diarrhea, abdominal pain, genital candidiasis, and pain at the site of injection, developed in 4% of the patients given cefodizime. Side effects, such as vertigo, genital candidiasis, fatigability, and diarrhea, developed in 4% of the patients treated with cefotaxime. In both groups of patients, the side effects were mild and transient. Cefodizime and cefotaxime are safe and effective agents in the treatment of uncomplicated urogenital gonorrhea.

Cefotaxime

Host range of the conjugative 25.2-megadalton tetracycline resistance plasmid from Neisseria gonorrhoeae and related species.

High-level tetracycline resistance in strains of Neisseria gonorrhoeae, Neisseria meningitidis, Kingella denitrificans, and Eikenella corrodens has recently been described. The resistance in each species is due to the acquisition of 25.2-megadalton conjugative plasmids that carry the tetracycline resistance determinant TetM. We examined the ability of commensal Neisseria species to serve as recipients in conjugation for these new plasmids. Most of the recipients (n = 21) tested had detectable conjugation frequencies (greater than 10(-9] with one or more of the donor strains. Transfer was not detected in Branhamella catarrhalis. Transconjugants were able to maintain the plasmids and act as donors in subsequent matings.

Conjugation, Genetic

Transfer of beta-lactamase plasmids from Neisseria gonorrhoeae to Neisseria meningitidis and commensal Neisseria species by the 25.2-megadalton conjugative plasmid.

Two highly tetracycline-resistant, beta-lactamase-producing Neisseria gonorrhoeae strains were used as donors for conjugation with Neisseria meningitidis and commensal Neisseria species. We found that both strains were able to transfer the 4.4- and 3.2-megadalton beta-lactamase plasmids to the recipients tested, with frequencies between 10(-1) and 10(-9).

Conjugation, Genetic