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J Fierer

Publications and source records attributed to J Fierer.

At least 55 records · Page 3Linked to original sources
PubMed1992

Biliary concentrations of fluconazole in a patient with candidal cholecystitis: case report.

A patient with acute cholecystitis due to Candida albicans and Candida parapsilosis was treated with a percutaneous cholecystostomy and daily intravenous fluconazole. Fluconazole levels in serum and bile were measured by gas chromatography. Fluconazole levels in the bile were equal to those in the blood for the first 8 hours after a dose and were slightly higher than serum levels after that. Bile levels after an oral dose of fluconazole were 15% higher than levels achieved after intravenous administration of the drug. The infection was cured after 2 weeks of treatment. This experience suggests that sufficient fluconazole is excreted in the bile to be effective for treatment of biliary infections due to susceptible yeasts.

Aged↗
PubMed1992

Salmonella typhimurium bacteremia: association with the virulence plasmid.

Virulence plasmids, which are found in a small number of Salmonella serotypes, greatly enhance the extraintestinal growth of salmonellae and lower the LD50 by 2-5 logs in experimental murine infections. To determine if virulence plasmids are important in the pathogenesis of Salmonella bacteremia in humans, blood and fecal isolates of Salmonella typhimurium from California were examined for the presence of a virulence plasmid. Colony blots were done using a labeled probe made from the highly conserved EcoRI fragment of the Salmonella dublin virulence plasmid. A total of 42% of the fecal and 76% of the blood isolates hybridized with the probe. This is the first evidence that the virulence plasmid is important in the pathogenesis of Salmonella bacteremia in humans.

Bacteremia↗
PubMed1992

The Salmonella typhimurium virulence plasmid complement resistance gene rck is homologous to a family of virulence-related outer membrane protein genes, including pagC and ail.

A fragment of the Salmonella typhimurium virulence plasmid containing the rck locus, when cloned in the recombinant cosmid pADE016, was shown previously to confer high-level complement resistance on both rough and smooth Escherichia coli, Salmonella minnesota, and S. typhimurium and was associated with the production of an outer membrane protein. We determined the nucleotide sequence of the fragment containing the rck locus. Mutations in the two major open reading frames confirmed that the complement resistance mediated by pADE016 was due to a single 555-bp rck gene encoding a 17-kDa outer membrane protein. Analysis of the rck gene revealed that the Rck outer membrane protein consisted of 185 amino acid residues, with a calculated postcleavage molecular mass of 17.4 kDa. Rck is homologous to a family of outer membrane proteins expressed in gram-negative bacteria, two of which have been associated with virulence-related phenotypes: PagC, required by S. typhimurium for survival in macrophages and for virulence in mice; and Ail, a product of the Yersinia enterocolitica chromosome capable of mediating bacterial adherence to and invasion of epithelial cell lines. Rck, most closely related to PagC, represents the third outer membrane protein in this five-member family with a distinct virulence-associated phenotype.

Amino Acid Sequence↗
PubMed1992

Characterization of translation termination mutations in the spv operon of the Salmonella virulence plasmid pSDL2.

The spv region of the Salmonella virulence plasmids consists of five genes located on an 8-kb fragment previously shown to be essential for virulence in mice. Four structural genes, spvABCD, form an operon that is transcriptionally activated by the spvR gene product in the stationary phase of growth. The role of the individual spv genes in the virulence phenotype was tested by isolating translation termination linker insertions in each gene. Analysis of proteins synthesized in minicells identified each of the spvABCD gene products and confirmed the dependence of spv structural gene expression on the SpvR regulatory protein. The oligonucleotide insertions in spvA, -B, and -C were shown to be nonpolar. Virulence testing indicated that the SpvB protein, regulated by SpvR, is essential for Salmonella dublin to cause lethal disease in mice. Inserts in spvC and spvD were unstable in vivo for unknown reasons, but these mutants still killed mice at slightly higher inocula. Abolition of spvA had no effect on virulence in this system.

Bacterial Proteins↗
PubMed1992

Mechanism of resistance to complement-mediated killing of bacteria encoded by the Salmonella typhimurium virulence plasmid gene rck.

We find that pADEO16, a recombinant cosmid carrying the rck gene of the Salmonella typhimurium virulence plasmid, when cloned into either rough or smooth Escherichia coli and Salmonella strains, confers high level resistance to the bactericidal activity of pooled normal human serum. The rck gene encodes a 17-kD outer membrane protein that is homologous to a family of virulence-associated outer membrane proteins, including pagC and Ail. Complement depletion, C3 and C5 binding, and membrane-bound C3 cleavage products are similar in strains with and without rck. Although a large difference in C9 binding was not seen, trypsin cleaved 55.7% of bound 125I-C9 counts from rough S. typhimurium with pADEO16, whereas only 26.4% were released from S. typhimurium with K2011, containing a mutation in rck. The majority of C9 extracted from rck strain membranes sediments at a lower molecular weight than in strains without rck, suggesting less C9 polymerization. Furthermore, SDS-PAGE analysis of gradient peak fractions indicated that the slower sedimenting C9-containing complexes in rck strains did not contain polymerized C9 typical of the tubular membrane attack complex. These results indicate that complement resistance mediated by Rck is associated with a failure to form fully polymerized tubular membrane attack complexes.

Antibodies, Monoclonal↗
PubMed1991

Synergistic effect of human lysozyme plus ampicillin or beta-lysin on the killing of Listeria monocytogenes.

Although ampicillin is often only bacteriostatic for Listeria monocytogenes in vitro, serum from ampicillin-treated patients was bactericidal. The bactericidal effect of serum was partly removed by bentonite, Seitz-filtration, and addition of FeCl3, suggesting it is mediated by lysozyme and beta-lysin. Partly purified human beta-lysin plus purified human lysozyme or either protein plus ampicillin were bactericidal for L. monocytogenes. Hen egg white lysozyme was not active. Lysozyme and beta-lysin were not synergistic with tetracycline, trimethoprim/sulfamethoxazole, or chloramphenicol. Thus, lysozyme and beta-lysin may play a role in the natural resistance to L. monocytogenes, and these serum proteins could contribute to the effectiveness of ampicillin in vivo.

Ampicillin↗
PubMed1991

Human infection with Salmonella dublin.

Twenty-seven cases of human infection with Salmonella dublin were identified over a 12-year period at the University of California at San Diego-affiliated hospitals. Important epidemiologic risk factors were the ingestion of unpasteurized dairy products or treatment with nutritional therapy that included raw calf-liver extracts. Nearly all patients had underlying chronic diseases. Like Salmonella choleraesuis, S. dublin infections were associated with a high incidence of bacteremia (91%), metastatic sites of infection (30%), and mortality (26%) relative to other non-typhoidal Salmonellae. This pattern of disease expression may be related to a plasmid-encoded virulence factor common to both of these organisms.

Adult↗
PubMed1991

Molecular analysis of the virulence locus of the Salmonella dublin plasmid pSDL2.

The virulence properties of various non-typhoid Salmonella serotypes depend on the presence of large plasmids 60-100 kb in size. We have shown previously that the virulence region on the 80 kb plasmid pSDL2 of Salmonella dublin Lane maps within a 14kb SalI fragment. In this report we show that an 8.2 kb region within this fragment is sufficient to express lethal disease in BALB/c mice. Sequence analysis of this segment revealed six sequential open reading frames designated vsdA-F, which encode putative proteins of 13-65kDa. Deletion analysis and location of Tn5-oriT inserts which abolish virulence suggest that vsdA, vsdC, vsdD and vsdE are essential for virulence expression. Downstream of vsdF we discovered a locus involved in stable plasmid maintenance. Deletion of that region resulted in plasmid multimerization and instability.

Amino Acid Sequence↗
PubMed1991

Genetic analysis of homology between the virulence plasmids of Salmonella dublin and Yersinia pseudotuberculosis.

Two segments within the virulence region of Salmonella dublin plasmid pSDL2 that were homologous to regions on Yersinia pseudotuberculosis plasmid pIB1 were located with regard to the four known genes (vsdA, vsdB, vsdC, and vsdD) of pSDL2. One segment mapped upstream of vsdA within an insertion element related to IS630 of Shigella sonnei; the second was confined to a 45-bp sequence containing an inverted repeat between vsdC and vsdD. On pIB1, both areas were located in an intergenic region upstream of yopH. These results indicate that the homology between Yersinia and Salmonella virulence plasmids is not located within structural genes.

Amino Acid Sequence↗
PubMed1991

Growth regulation of a Salmonella plasmid gene essential for virulence.

The Salmonella dublin plasmid gene vsdC is essential for virulence. We have constructed a vsdC-lacZ translational fusion to demonstrate that vsdC is selectively expressed during the stationary phase of bacterial cell growth. This pattern of expression has been confirmed by mRNA hybridization studies. Carbon starvation is able to induce vsdC expression by limiting bacterial growth. The expression of vsdC is dependent upon an upstream gene, vsdA, whose gene product possesses significant amino-terminus homology with the LysR family of transcriptional activator proteins. We have further demonstrated that vsdC expression is not dependent upon the known Salmonella chromosomal virulence regulatory loci ompR, phoP, and cya-crp and that vsdC can be expressed in a range of nontyphoidal Salmonella serovars, including some serovars in which introduction of the virulence plasmid does not confer mouse virulence. The vsd system provides a model for the study of transcriptional activation, a basis for the development of new expression vectors, and a novel mechanism of virulence gene regulation. Bacterial growth limitation within the phagosomes of host phagocytic cells may be the environmental signal inducing plasmid-mediated virulence gene expression in salmonellae.

Blotting, Northern↗
PubMed1991

Type B Haemophilus influenzae-induced otitis media in the mouse.

To characterize the middle and inner ear cellular inflammatory responses to otitis media using immunohistochemical methods, we inoculated type B Haemophilus influenzae into the middle ears of healthy adult BALB/c mice. Mac-1+ neutrophils and macrophages appeared in the middle ear at 3 days. Lyt-1+ T cells and Lyt-2+ T suppressor/cytotoxic cells entered the middle ear mucosa on days 7 and 14. IgG+ and IgM+ T cells were present at all time points, with IgA+ lymphocytes forming the majority of mucosal immunoglobulin-bearing cells at 2 weeks. The cochlear scala tympani contained Lyt-1+ and Mac-1+ cells and two endolymphatic sacs stained diffusely with anti-IgA and -IgG antibodies. Lyt-1/L3T4+ T lymphocytes greatly outnumbered B lymphocytes, suggesting that helper/inducer T cells play a more important role in acute otitis media than has been recognized. Inner ear changes occurred after a single episode of otitis media.

Animals↗
PubMed1990

Homologous DNA sequences on the virulence plasmids of pathogenic Yersinia and Salmonella dublin lane.

Yersinia and Salmonella harbour plasmids that encode traits important for virulence, enabling both pathogenic genera to survive and grow in cells of the reticulo-endothelial organs during systemic infections. We have detected DNA homology between the Salmonella dublin virulence plasmid pSDL2 and the plasmids of the pathogenic Yersinia species pestis, pseudotuberculosis, and enterocolitica. Three regions of pSDL2 were found to share homology with the virulence plasmid pIB1 of Yersinia pseudotuberculosis. Two separate hybridizing segments mapped within the previously characterized 6.4 kb vir region of pSDL2 in the SalI B fragment. The third homologous region involved the replicon of pIB1, which hybridized to the SalI C2 fragment of pSDL2. The virulence plasmid pCD1 from Y. pestis showed similar homology with the three regions of pSDL2. Homologies to the vir and SalI C2 regions of pSDL2 were also found on plasmids from Yersinia enterocolitica serotypes 0:9, 0:3 and 0:5, 27. The discovery of separate homologous regions on the virulence plasmids of Salmonella and Yersinia suggests a distant evolutionary relationship.

Blotting, Southern↗
PubMed1990

Comparison of fluconazole and SDZ89-485 for therapy of experimental murine coccidioidomycosis.

We infected mice with arthroconidia of Coccidioides immitis by intraperitoneal injection and 48 h later treated them with either oral fluconazole or SDZ89-485, a new triazole. Both drugs completely inhibited fungal growth when administered at a dose of 50 mg/kg of body weight twice a day, but only SDZ89-485 was fully inhibitory at a dose of 5 mg/kg twice a day. In a second experiment, treatment with SDZ89-485 was delayed until 8 days after infection to allow infection to be well established before treatment. Both 5 and 50 mg/kg twice a day were effective regimens, which establishes that SDZ89-485 has activity against spherules in vivo. Mice that received fluconazole (50 mg/kg twice a day) had a peak level in blood of 60 micrograms/ml 1 h after a dose, but no measurable amount was found after 12 h. SDZ89-485 was more slowly absorbed, reaching a peak level in blood of 14 micrograms/ml at 12 to 15 h after a dose of 50 mg/kg. We conclude that SDZ89-485 is more effective than fluconazole as treatment for experimental systemic coccidioidomycosis in mice, even though fluconazole achieves higher peak levels in blood.

Animals↗
PubMed1990

Successful treatment using gentamicin liposomes of Salmonella dublin infections in mice.

Gentamicin entrapped within stable multilamellar liposomes was used to treat mice after they were infected per os with Salmonella dublin. Of 10 mice, 8 survived after a single intravenous (i.v.) injection of 2 mg of gentamicin liposomes per kg compared with 0 of 10 treated with the same amount of free gentamicin. All mice survived after treatment with a single i.v. or intraperitoneal injection of 20 mg of gentamicin liposomes per kg, whereas that dose of free drug was completely ineffective and caused neuromuscular paralysis when injected rapidly i.v. In mice treated with gentamicin liposomes, there was a steady decrease in the number of salmonellae in spleens for 2 weeks after treatment. High concentrations of gentamicin were present in the spleen for at least 10 days after treatment. Although gentamicin was not detected in the mesenteric lymph nodes of mice treated with gentamicin liposomes, bacterial counts in the nodes also decreased over time. Small numbers of bacteria remained viable in the mesenteric lymph nodes and Peyer's patches but not in the spleens of mice treated with 20 to 80 mg/kg. Mice treated with doses of gentamicin liposomes as high as 80 mg/kg showed only a transient increase in blood urea nitrogen and no rise in serum creatinine. These results confirm that gentamicin in liposomes is less toxic in mice than is free gentamicin and is extremely effective therapy for disseminated Salmonella infections in mice.

Animals↗
PubMed1990

Correlation between the presence of sequences homologous to the vir region of Salmonella dublin plasmid pSDL2 and the virulence of twenty-two Salmonella serotypes in mice.

Large plasmids encoding important virulence properties have been found in several Salmonella serotypes. We have studied the relationship between the presence of a highly conserved 4-kilobase (kb) EcoRI fragment from the plasmid virulence region and pathogenicity for mice of 53 isolates representing 22 serotypes of Salmonella. Only strains possessing the homologous 4-kb region were virulent for mice. In addition, we transferred the virulence plasmid from S. dublin into nine different serotypes, including S. typhi and S. paratyphi A, that lack a native virulence plasmid. Only S. heidelberg and S. newport were rendered mouse virulent by the introduction of the S. dublin plasmid. This study demonstrates that plasmid-mediated virulence sequences are required for Salmonella virulence in mice, but many strains, including the agents of human typhoid fever, also lack chromosomal genes necessary to produce lethal systemic disease in mice. Since all the major Salmonella strains that are host-adapted to animals carry virulence plasmids, it appears that these plasmids are important in mediating systemic infection in animals and may contribute to septicemic, nontyphoid salmonellosis in humans.

Animals↗
PubMed1990

Fluconazole in the treatment of persistent coccidioidomycosis.

Fluconazole is one of the new antifungal triazoles undergoing clinical trials. We used fluconazole at a dose of 50 or 100 mg/day in an open trial for the treatment of patients with persistent coccidioidomycosis. Fourteen patients were enrolled and treated for a mean of 13 +/- 7 months. Two failed to respond. Of the 12 who responded, one reactivated while being treated, and one died of myocardial infarction after successful treatment of his fungal infection; six had relapses from nine days to 15 months after treatment was stopped. Only four patients are asymptomatic at a mean of 14 +/- 3 months after cessation of treatment. Fluconazole is well tolerated at this dose. In view of its low toxicity, the partial clinical efficacy observed, and the high recurrence rate of chronic coccidioidal infection, it would be justified to try higher doses.

Adult↗