bluuebunny/crossref_metadata_2025 huggingface dataset:
Cloning and expression in Escherichia coli of a gene encoding nonenzymatic chloramphenicol resistance from Pseudomonas aeruginosa
Authors: Burns J L , Rubens C E , Mendelman P M , Smith A L
Published: June 2012

High-level chloramphenicol resistance in Pseudomonas aeruginosa may be due to enzymatic inactivation, ribosomal mutation, or a permeability barrier. We investigated the nonenzymatic resistance mechanism encoded by Tn1696, a transposon found in P. aeruginosa. A 1-megadalton DNA fragment from Tn1696 was cloned which mediated expression of chloramphenicol resistance in Escherichia coli. Comparison of the effects of chloramphenicol on in vitro translation revealed no difference between the susceptible recipient strain and the resistant transformant containing the cloned gene. The rate of chloramphenicol uptake was slower in the resistant strain, suggesting a permeability barrier to the antibiotic. In addition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of outer membranes demonstrated the absence of a 50,000-dalton protein in the resistant strain. DNA homology was evident between Tn1696 and chloramphenicol-resistant isolates of Haemophilus influenzae possessing altered outer membrane permeability. We conclude that chloramphenicol resistance encoded by Tn1696 is due to a permeability barrier and hypothesize that the gene from P. aeruginosa may share a common ancestral origin with these genes from other gram-negative organisms.
Enoxacin compared with vancomycin for the treatment of experimental methicillin-resistant Staphylococcus aureus endocarditis
Authors: Gilbert M , Boscia J A , Kobasa W D , Kaye D
Published: June 2012

Enoxacin administered orally was compared with vancomycin administered intravenously for the treatment of experimental methicillin-resistant Staphylococcus aureus endocarditis. The MICs and MBCs of both enoxacin and vancomycin for an inoculum of 5.0 X 10(5) CFU of the methicillin-resistant S. aureus strain per ml were 1.56 microgram/ml. With an inoculum of 10(8) CFU/ml, enoxacin at 6 micrograms/ml and vancomycin at 180 micrograms/ml resulted in similar decreases in numbers of methicillin-resistant S. aureus in broth. Methicillin-resistant S. aureus endocarditis in rabbits was treated with enoxacin at 100 mg/kg orally every 12 h or vancomycin at 30 mg/kg intravenously every 12 h for 3 or 5 days. Enoxacin treatment for 3 or 5 days and vancomycin treatment for 5 days significantly reduced bacterial counts of vegetations compared with those in untreated control rabbits after 1 day of infection. Bacterial counts of vegetations after vancomycin treatment for 3 days did not differ significantly from those of untreated controls. Bacterial counts of vegetations in the four therapeutic groups did not differ significantly from one another. In uninfected rabbits single doses of vancomycin at 30 mg/kg administered intravenously achieved much higher concentrations in serum than did single doses of enoxacin at 100 mg/kg administered orally. Enoxacin had an elimination half-life in serum that was approximately 1.5 times longer than that of vancomycin. This study demonstrated that enoxacin administered orally is as effective as vancomycin administered intravenously for the treatment of experimental methicillin-resistant S. aureus endocarditis.
Canavanine resistance in Cryptococcus neoformans
Authors: Polacheck I , Kwon-Chung K J
Published: June 2012

All of the isolates of Cryptococcus neoformans var. gattii which we tested were resistant to greater than or equal to 3.5 mM canavanine. All of the serotype D isolates and 28% of the serotype A isolates of C. neoformans var. neoformans tested were susceptible to less than or equal to 18 microM canavanine, whereas the remaining 72% of the serotype A isolates were as resistant as the C. neoformans gattii isolates. In the naturally resistant isolates, the mechanism of resistance appeared to be decomposition of canavanine to a nontoxic product. However, in a resistant mutant derived from a naturally susceptible isolate, the mechanism of resistance was an impaired uptake system for canavanine. The toxic effect of canavanine in Cryptococcus results from the incorporation of canavanine into the protein component that is essential for the synthesis of proteins and RNAs.
Interaction of polycationic antibiotics with Pseudomonas aeruginosa lipopolysaccharide and lipid A studied by using dansyl-polymyxin
Authors: Moore R A , Bates N C , Hancock R E
Published: June 2012

A fluorescent derivative of polymyxin B (dansyl-polymyxin) was used to study the interaction of polycations with lipopolysaccharide (LPS) and lipid A from Pseudomonas aeruginosa. Dansyl-polymyxin became bound to LPS and lipid A sites, including Mg2+-binding sites, resulting in a 20-fold enhancement of fluorescence. A Hill plot of the binding data showed that the binding of dansyl-polymyxin to LPS was cooperative (n = 1.98) and of high affinity (S0.5 = 0.38 microM). The maximal binding capacity of LPS was approximately four molecules of dansyl-polymyxin per mol of LPS. The dansyl-polymyxin interaction with lipid A displayed similar kinetics (n = 2.26; S0.5 = 0.38 microM), and the maximal binding capacity was approximately 2 mol of dansyl-polymyxin per mol of lipid A. A variety of polycationic compounds, including gentamicin, streptomycin, and polymyxin B, as well as Mg2+, were able to displace dansyl-polymyxin bound to LPS or to lipid A. Marked differences both in terms of the degree of displacement and in terms of the amount of competing polycation required to displace a given amount of dansyl-polymyxin were observed. Addition of excess polymyxin B resulted in displacement of all of the dansyl-polymyxin, demonstrating that only polymyxin-binding sites were being probed. Our data demonstrate that polymyxin B binds to multiple sites on LPS, including sites which bind aminoglycoside antibiotics and other polycationic compounds.
Successful treatment and prevention of murine Pneumocystis carinii pneumonitis with 4,4'-sulfonylbisformanilide
Authors: Hughes W T , Smith B L , Jacobus D P
Published: June 2012

Pneumocystis carinii pneumonitis was prevented in 0, 50, 100, and 100% of immunosuppressed rats given doses of 0.5, 5.0, 25.0, and 125.0 mg/kg (body weight) per day, respectively, of 4,4'-sulfonylbisformanilide (DFD). Therapeutic efficacy was demonstrated with DFD at 25.0 mg/kg per day, and when this dose was combined with trimethoprim, the combination was as effective as trimethoprim-sulfamethoxazole, which has been proven to be effective in the treatment of murine and human P. carinii pneumonitis.
A novel type of resistance plasmid in Haemophilus influenzae
Authors: Piffaretti J C , Camponovo T , Borer-Bigliardi B , Arini A
Published: June 2012

Resistance plasmids of a novel type were found in two Haemophilus influenzae clinical isolates. pPJ301 and pPJ302 are 10.0 kilobases in size, carry a Tn2-like transposable element, and are related only by their common beta-lactamase genes to the other two types of resistance plasmids known to occur in H. influenzae.
Combined action of chloramphenicol and ampicillin on chloramphenicol-resistant Haemophilus influenzae
Authors: Mackenzie A M , Chan F T
Published: June 2012

The interaction of ampicillin and chloramphenicol on three ampicillin-susceptible, chloramphenicol-resistant strains of Haemophilus influenzae was studied by checkerboard testing with subcultures, time-kill experiments, and a disk method. In all three strains there was inhibition of the bactericidal action of ampicillin by chloramphenicol at concentrations close to the MIC (10 micrograms/ml). This chloramphenicol concentration was close to that which might be achieved in cerebrospinal fluid during treatment for meningitis and was in the bactericidal range for chloramphenicol-susceptible organisms. It is suggested however that in the initial treatment of meningitis caused by ampicillin-susceptible, chloramphenicol-resistant strains, inhibition of the action of ampicillin by chloramphenicol may represent a clinical risk.
Molecular evolution, species distribution, and clinical consequences of an endemic aminoglycoside resistance plasmid
Authors: Mayer K H , Hopkins J D , Gilleece E S , Chao L , O'Brien T F
Published: June 2012

During the first 6 years after appearing in one hospital, a 92-kilobase conjugative plasmid, pBWH1, which encoded resistance to chloramphenicol and sulfonamides and determined TEM-1 beta-lactamase and 2''-aminoglycoside nucleotidyltransferase, underwent a variety of molecular changes. It was most prevalent initially in isolates of Klebsiella pneumoniae, then in isolates of Serratia marcescens, and finally, after nearly disappearing, in isolates of Enterobacter cloacae. Evolutionary changes in the plasmid did not account for its shifts in species distribution, since the original molecule was found in isolates of each species. The late resurgence of pBWH1 occurred after a copy of its original molecule entered a distinctive ornithine decarboxylase-negative strain of E. cloacae, new to the hospital. The resulting transconjugant strain, chromosomally resistant to topical silver salts and to cephalosporins, and with the addition of pBWH1-encoded aminoglycoside resistance, spread in the hospital by causing an outbreak of sepsis in the burn unit, where these were commonly used antibacterial agents. Thus, an endemic plasmid became prevalent in a new host species because one of its genes supplemented the fitness of an uncommon strain of the species for a particular clinical niche.
Evaluation of antibacterial agents in a high-volume bovine polymorphonuclear neutrophil Staphylococcus aureus intracellular killing assay
Authors: Sanchez M S , Ford C W , Yancey R J
Published: June 2012

A bovine polymorphonuclear leukocyte (PMN) monolayer system was used to determine the ability of different antibiotics to kill surviving intracellular Staphylococcus aureus. The following classes of antimicrobial agents were tested in this high-volume assay procedure: aminocyclitol, beta-lactam, coumarin, lincosaminide, macrolide, naphthalenic ansamycin, paulomycin, peptide, quinolone, and tetracycline. The activities of these compounds were compared with those of positive (rifampin), negative (cloxacillin), and non-antibiotic-treated controls. Only oxytetracycline, the ansamycins (rifampin, rifamycin SV, streptovaricin A, C, and D), paulomycin, and paldimycin caused a significant reduction in the viable count of intracellular S. aureus. Of these, however, the intracellular killing by the streptovaricins was directly related to the cytotoxicity (as determined by trypan blue exclusion) of these compounds for the PMNs. Although the paulomycins were cytotoxic for the PMNs, the cytotoxic and the intracellular killing activity of these new compounds could be distinguished. The relevance of these results to the therapeutic effectiveness of these antibiotics in the treatment of bovine staphylococcal mastitis is discussed.
Genetic and biochemical characterization of norfloxacin resistance in Escherichia coli
Authors: Hooper D C , Wolfson J S , Souza K S , Tung C , McHugh G L , Swartz M N
Published: June 2012

In Escherichia coli the frequency of spontaneous single-step mutation to high levels of resistance to the newer 4-quinolone agent norfloxacin was confirmed to be over 300-fold lower than that to the older agent nalidixic acid. Serial passage on incremental concentrations of drug was necessary to produce mutants highly resistant to norfloxacin. Genetic analysis of one such highly resistant strain identified two mutations conferring drug resistance. One mutation, nfxA, mapped around 48 min on the E. coli genetic map and was shown to be an allele of gyrA by studies demonstrating an increased drug resistance of DNA gyrase reconstituted with the gyrase A subunit isolated from the mutant strain. These findings also identified the DNA gyrase A subunit as a target of norfloxacin. The second mutation, nfxB, mapped between 20 and 22 min was associated with additional resistances to tetracycline, chloramphenicol, and cefoxitin and with decreases in outer membrane porin protein OmpF. The nfxA and nfxB mutations together accounted for most, but not all, of the norfloxacin resistance phenotype of this strain.
Comparison of the in vitro and in vivo activity of the bis-triazole derivative UK 49,858 with that of amphotericin B against Histoplasma capsulatum
Authors: Kobayashi G S , Travis S , Medoff G
Published: June 2012

The antifungal activity of UK 49,858, a difluorophenyl bis-triazole derivative, was evaluated in vitro against seven strains of Histoplasma capsulatum and in vivo in AKR and C57BL/6 murine models of histoplasmosis. UK 49,858 had a lower toxicity for AKR and C57BL/6 mice than amphotericin B did. The therapeutic index of UK 49,858 was 4.3 for AKR mice and 7.1 for C57BL/6; with amphotericin B it was 2 for both mouse strains. Given orally, UK 49,858 compared favorably with amphotericin B given intraperitoneally in either AKR or C57BL/6 mice infected with H. capsulatum.
Resistance to mercury and to cadmium in chromosomally resistant Staphylococcus aureus
Authors: Witte W , Green L , Misra T K , Silver S
Published: June 2012

Apparently chromosomally located mercury resistance determinants in five methicillin-resistant Staphylococcus aureus strains of different geographical origin were structurally homologous to plasmid-located mercury resistance determinants in S. aureus. These were all located on a 6.3-kilobase (kb) Bg/II fragment, as evident from Southern hybridization experiments with the 6.3-kb Bg/II fragment of plasmid pI258 as the probe. These methicillin-resistant S. aureus strains exhibited similar phage susceptibility patterns and biochemical reactions. They differed, however, in the DNA location of the mercury resistance determinants, as evidenced by neighboring cleavage sites for restriction endonucleases EcoRI, HindIII, and PstI. In an environmental (nonhospital) strain in which mercury resistance was also apparently chromosomally conferred, these determinants were also homologous to pI258 DNA, but they were located on a 6.6-kb Bg/II fragment. Cadmium resistance determinants in the five methicillin-resistant S. aureus strains and the environmental S. aureus strain were not similar to the known plasmid-located determinants cadA and cadB. Cd2+ resistance was based on an efflux mechanism for Cd2+. However, no parallel resistance to zinc was conferred. The 3.2-kb XbaI-Bg/II fragment obtained from plasmid pI258 and used as a cadA-specific probe did not hybridize to total DNA digests of the strains with apparently chromosomally determined cadmium resistance.
Purification and properties of a beta-lactamase produced by Branhamella catarrhalis
Authors: Yokota E , Fujii T , Sato K , Inoue M , Mitsuhashi S
Published: June 2012

A beta-lactamase from Branhamella catarrhalis was purified by column chromatography. The purified enzyme hydrolyzed penicillins, such as ampicillin, carbenicillin, and piperacillin, more rapidly than cephalosporins. Furthermore, the enzyme hydrolyzed cefotaxime and cefmenoxime. The molecular weight of the enzyme was 33,000. The pI was 5.4.
Incorporation of 1-beta-D-arabinofuranosylcytosine into DNA and mutagenesis of herpes simplex virus type 1
Authors: Bubley G J , Crumpacker C S , Schnipper L E
Published: June 2012

To investigate the association between incorporation of 1-beta-D-arabinofuranosylcytosine (ara-C) into herpes simplex virus type 1 (HSV-1) DNA and mutagenesis at selected genetic loci, we grew virus in the presence or absence of ara-C and compared the mutation frequencies. Both the forward mutation frequency of HSV-1 (KOS) to the tk- phenotype and the reversion frequency of a temperature-sensitive mutant to the non-temperature-sensitive (ts+) phenotype were significantly increased following growth in ara-C. When the same viruses were grown in an inhibitor of the HSV-1 DNA polymerase that is not incorporated into DNA, no significant increase in the frequency of tk- or ts+ particles was observed. Furthermore, HSV-1 araAr, a mutant resistant to ara-C on the basis of reduced incorporation of the analog into viral DNA, did not demonstrate an increase in the number of tk- particles following growth in ara-C. These results suggest that mutagenesis of HSV-1 following growth in ara-C correlates with incorporation of the nucleoside analog into viral DNA.
Failure of topical acyclovir in ointment to penetrate human skin
Authors: Freeman D J , Sheth N V , Spruance S L
Published: June 2012

Topical acyclovir (ACV) in polyethylene glycol (PEG) ointment has been disappointing in the treatment of recurrent herpes simplex virus infections in immunocompetent patients. To investigate the possible role of poor drug delivery from this formulation, we studied the penetration of ACV through excised human skin from three vehicles; PEG ointment, modified aqueous cream, and dimethyl sulfoxide. A second antiviral agent, idoxuridine, was studied in the same formulations, and drug delivery through excised guinea pig skin was also assessed for comparison. The delivery of ACV from PEG ointment was very slow for both human and guinea pig skin (drug flux, 0.055 and 0.047 microgram/cm2 per h, respectively). Formulation of ACV in modified aqueous cream and in dimethyl sulfoxide resulted in an 8- and 60-fold increase, respectively, in the flux of ACV through human skin. Idoxuridine behaved similarly to ACV in the three vehicles. The poor clinical results seen with topical use of ACV ointment may be due in part to retarded drug delivery from this formulation.
Imipenem therapy of experimental Staphylococcus epidermidis endocarditis
Authors: Berry A J , Johnston J L , Archer G L
Published: June 2012

Imipenem was evaluated for its activity against Staphylococcus epidermidis in vitro and in a rabbit model of endocarditis. The MBC for imipenem of 55 methicillin-resistant S. epidermidis isolates from patients with prosthetic valve endocarditis increased by eightfold or greater with increasing inoculum size; there was no inoculum-associated increase in the imipenem MBC for 20 methicillin-susceptible S. epidermidis isolates. Endocarditis was produced in rabbits with either a methicillin-susceptible or a methicillin-resistant S. epidermidis isolate to investigate the correlation in vivo of the in vitro inoculum effect for imipenem. Six days of imipenem treatment eradicated methicillin-susceptible S. epidermidis from vegetations of infected rabbits significantly better than no therapy but was less effective against methicillin-resistant S. epidermidis in this regard. Among methicillin-resistant S. epidermidis-infected rabbits, 6 days of imipenem therapy (i) was not significantly better than that of the control and was significantly worse than that of vancomycin in eradicating bacteria from infected vegetations and (ii) increased the frequency of imipenem-resistant subpopulations in infected vegetations. Resistant subpopulations were not seen in vegetations from untreated or imipenem-treated, methicillin-susceptible S. epidermidis-infected rabbits. Imipenem may not be effective therapy for serious human methicillin-resistant S. epidermidis infections.
Influence of inflammation on the efficacy of antibiotic treatment of experimental pyelonephritis
Authors: Meylan P R , Braoudakis G , Glauser M P
Published: June 2012

An acute exudative Escherichia coli pyelonephritis rat model was used to study the influence of progressive pyelonephritis on the efficacy of antibiotic treatment. In this model, transient ureteral obstruction after E. coli bladder inoculation induces early bacterial multiplication in the kidney parenchyma, and the bacterial counts peak by 48 h. The inflammatory response (assessed by the increase in kidney weight) is somewhat delayed, starting 36 h after inoculation and peaking by 72 h. Groups of rats received 4 doses over 48 h of saline, ceftriaxone (100 mg/kg), or ceftriaxone (100 mg/kg) plus gentamicin (4 mg/kg). These treatments were initiated 24, 36, 48, or 72 h after bladder inoculation. Antibiotic treatment started at 24 h was significantly more effective in reducing bacterial counts in the kidney parenchyma than at any later therapy onset. Only when started 24 h after inoculation was the synergistic combination of ceftriaxone plus gentamicin more effective in reducing bacterial counts than ceftriaxone alone. Ceftriaxone and ceftriaxone plus gentamicin regimens started at 24 h reduced significantly (by 42 and 55%, respectively) the incidence of acute exudative pyelonephritis when compared with the incidence in saline-treated controls. Early therapy onset (24 h) strikingly reduced the development of the inflammatory response. This reduction was less marked when antibiotic therapy was started at 36 h and no longer apparent when therapy onset was delayed up to 48 or 72 h. In conclusion, the efficacy of antibiotics in eradicating bacteria from the kidney parenchyma and in preventing acute exudative pyelonephritis was markedly hampered by the development of pyelonephritis.
Artifacts in dilution pharmacokinetic models caused by adherent bacteria
Authors: Haag R , Lexa P , Werkhäuser I
Published: June 2012

Liquid cultures of a Pseudomonas aeruginosa strain in Mueller-Hinton broth diluted at rates higher than the bacterial growth rate showed the expected decrease in CFU only for 1 to 2 h. Later the CFU started to increase. This phenomenon can be explained by a hypothesis that assumes that the bacteria multiply in two different compartments. From the first compartment, which comprises bacteria homogeneously distributed in the broth, cells are eliminated at a rate that is dependent on the dilution and growth rates. Concomitantly, the second compartment is formed as a nondilutable adherent population on the surface of the culture vessel. Eventually, only cells stemming from that population appeared in the medium and were subsequently diluted. This hypothesis can be described mathematically by a linear combination of two exponential functions. The calculated values fit the experimental data well. Because similar CFU versus time curves were also found with other strains, care should be taken in interpreting results of experiments performed in liquid cultures and evaluated in terms of CFU. One should bear in mind that within a liquid culture an adherent population may exist, which differs in size according to selective influences (dilution, addition of antibiotics, etc.). This may give rise to artificial and unexpected results.
Quaternary ammonium compounds efficiently inhibit Plasmodium falciparum growth in vitro by impairment of choline transport
Authors: Ancelin M L , Vial H J
Published: June 2012

Hemicholinium 3, decamethonium, and decyltrimethylammonium previously have been demonstrated to be efficient inhibitors, with 50% inhibitory concentrations of 4 X 10(-6), 10(-6), and 7 X 10(-7) M, respectively. We show that lengthening of the alkyl chain of decyltrimethylammonium by successive additions of two carbon atoms up to hexadecyltrimethylammonium results in a very low 50% inhibitory concentration of 5 X 10(-7) M for dodecyltrimethylammonium. Furthermore, hemicholinium 3 and decamethonium exerted their antiplasmodial activity, regardless of the developmental stage of the parasite, whereas decyltrimethylammonium was particularly lethal for the mature forms. After infected erythrocytes with radioactive choline were supplied, the determination of the water-soluble choline-containing compounds, as well as the assay of choline kinase activity, showed that the specific inhibition of phosphatidylcholine biosynthesis is related to the impairment of choline entry into erythrocytes. Thus, the impairment of the transport of choline, a natural polar head group of phospholipids, appears to be lethal for Plasmodium falciparum in vitro and could be a reasonable approach for a new malaria chemotherapy.
Dose ranging study of cefpimizole (U-63196E) for treatment of uncomplicated gonorrhea in men
Authors: Sandberg E T , Pegram P S , Roddy R E , Handsfield H H , Hampton K D , Shafran K M , Hook E W
Published: June 2012

We conducted a two-center dose ranging study to evaluate the efficacy, tolerance, and toxicity of cefpimizole, a new cephalosporin, in the treatment of uncomplicated gonorrhea in 96 males. Twelve patients at each center were treated intramuscularly with single doses of 1.0, 0.5, 0.25, and 0.125 g of cefpimizole. All urethral infections were cured at the highest dose, but lower doses produced progressively decreasing cure rates of 90% (0.5 g), 83% (0.25 g), and 71% (0.125 g). Treatment failures of rectal and pharyngeal infections occurred at the highest dose level. Geometric mean MICs for cefpimizole for successfully and unsuccessfully treated volunteers were 0.088 and 0.282 micrograms/ml, respectively. A prominent adverse effect was clinically significant pain at the injection site, which occurred in 57 (59%) of 96 patients. Results of the study demonstrate that cefpimizole offers no advantage over currently available antibiotics in the treatment of uncomplicated gonorrhea in men.
Light-dependent activity of the antitumor antibiotics ravidomycin and desacetylravidomycin
Authors: Greenstein M , Monji T , Yeung R , Maiese W M , White R J
Published: June 2012

The antitumor antibiotics ravidomycin and desacetylravidomycin were studied by the biochemical lambda prophage induction assay. In this assay, induction of the enzyme beta-galactosidase is measured as a specific indication of the ability of an agent to directly or indirectly damage DNA. Induction was observed only when these two antibiotics were irradiated with light in the presence of the indicator organism. Drug treated with light followed by incubation with the indicator organism in the dark did not cause induction. Light in both the near UV and visible wave length ranges activated these antibiotics; near UV and visible blue wavelengths were most effective, while 597-nm light was totally ineffective. The amount of induction caused by these drugs varied directly with the dosage of light provided. Bacterial growth inhibition, as well as cytotoxicity for a human colon carcinoma cell line, was also dramatically enhanced by light. These data suggest that ravidomycin and desacetylravidomycin are potent photosensitizing, DNA-damaging agents.
Enzymology and pathogenicity in mice of a herpes simplex virus type 1 mutant resistant to 2'-fluoro-2'-deoxy-1-beta-D-arabinofuranosyl-5-iodocytosine
Authors: Colacino J , Brownridge E , Greenberg N , Lopez C
Published: June 2012

The deoxypyrimidine nucleoside analog 2'-fluoro-2'-deoxy-1-beta-D-arabinofuranosyl-5-iodocytosine (FIAC) is a potent and selective inhibitor of herpes simplex virus type 1 in vitro (C. Lopez, K. A. Watanabe, and J. J. Fox, Antimicrob. Agents Chemother. 17:803-806, 1980). Isopycnographic analysis demonstrated that 1 microM FIAC inhibited herpes simplex virus DNA replication by more than 95% but inhibited cellular DNA replication by only 32%. Mutant herpes simplex virus type 1 selected resistant to FIAC displayed linked resistance to other nucleoside analogs, including arabinosylthymine and acyclovir. Lysates derived from Vero cells infected with FIAC-resistant virus showed markedly lower levels of thymidine kinase activity and were unable to phosphorylate selectively arabinosylthymine or FIAC, in contrast to lysates from cells infected with wild-type herpes simplex virus type 1. Finally, drug-resistant virus displayed a 6,000-fold decrease in pathogenicity when inoculated intraperitoneally into genetically susceptible A/J mice. These results indicate that resistance to deoxypyrimidine nucleoside analogs is due, at least in part, to alterations in viral thymidine kinase and is accompanied by decreased pathogenicity in vivo.
In vitro activity of the new macrolide antibiotic roxithromycin (RU 28965) against clinical isolates of Haemophilus influenzae
Authors: Jorgensen J H , Redding J S , Howell A W
Published: June 2012

The in vitro activity of the new macrolide antibiotic roxithromycin (RU 28965) was compared with the activities of five other orally absorbable antimicrobial agents against 100 clinical isolates of Haemophilus influenzae. Roxithromycin MICs were generally twofold to fourfold higher than those of erythromycin; the MIC for 90% of the strains for roxithromycin was 8 micrograms/ml.
Influence of medium and method on the in vitro susceptibility of Pseudomonas aeruginosa and other bacteria to ciprofloxacin and enoxacin
Authors: Blaser J , Dudley M N , Gilbert D , Zinner S H
Published: June 2012

Ciprofloxacin and enoxacin were two- to fourfold less active against Pseudomonas aeruginosa in calcium- and magnesium-supplemented broth compared with unsupplemented broth regardless of inoculum size, presence of serum, or use of inhibitory or bactericidal endpoints (P less than 0.01). The effect of cation supplementation was less pronounced and less consistent for Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus.
Pharmacokinetic study of lenampicillin (KBT-1585) in healthy volunteers
Authors: Saito A , Nakashima M
Published: June 2012

The pharmacokinetic properties of lenampicillin (KBT-1585), a new ampicillin ester, were investigated in 41 healthy volunteers. The maximum concentration of ampicillin in serum after oral administration of 400 mg of lenampicillin was 6.5 micrograms/ml at 0.70 h, and that after a equimolar dosage of ampicillin was 2.9 micrograms/ml at 0.87 h.
In vitro activity of clofazimine against rapidly growing nonchromogenic mycobacteria
Authors: Ausina V , Condom M J , Mirelis B , Luquin M , Coll P , Prats G
Published: June 2012

The in vitro activity of clofazimine against 80 isolates of rapidly growing nonchromogenic mycobacteria was studied by an agar dilution method. The drug inhibited 96% of strains tested at concentrations less than or equal to 1 microgram/ml, and it appears to be an agent of potential efficacy against Mycobacterium fortuitum and M. chelonae.
Antibacterial activity of GO 10213, a nitroimidazole derivative
Authors: Hof H , Ströder J
Published: June 2012

Compound GO 10213, a 5-nitroimidazole substituted at the 2 position, is more active against aerobic, microaerophilic, and anaerobic bacteria than metronidazole. The MICs for Salmonella typhimurium (2 to 128 micrograms/ml), Campylobacter spp. (0.06 to 16 micrograms/ml) and Bacteroides spp. (0.03 to 0.25 micrograms/ml) are definitely lower than those of metronidazole. Niridazole, a 5-nitrothiazole, is still more active.
Role of a novel antidiarrheal agent, BW942C, alone or in combination with trimethoprim-sulfamethoxazole in the treatment of traveler's diarrhea
Authors: Ericsson C D , Johnson P C , DuPont H L , Morgan D R
Published: June 2012

The efficacy of BW942C, a novel enkephalinlike pentapeptide antidiarrheal agent, was compared with the efficacy of trimethoprim-sulfamethoxazole (TMP-SMX) and the combination of the two agents in a placebo-controlled trial of the 72-h treatment of acute diarrhea. Subjects with diarrhea but without bloody stools or fever greater than 102 degrees F (38.9 degrees C) were enrolled. Administered to 134 U.S. adults with diarrhea that developed shortly after their arrival in Guadalajara, Mexico, BW942C was more efficacious than TMP-SMX in relieving diarrhea and cramps in the first 12 h of therapy, especially among subjects with diarrhea caused by enterotoxigenic E. coli. In the BW942C treatment group, 25% of subjects eventually took additional therapy because their diarrhea did not respond to BW942C alone. Neurological side effects such as dizziness and light-headedness occurred more frequently among BW942C-treated subjects. Therapy for 3 days with TMP-SMX provided lasting relief comparable with previously reported 5-day therapy. Use of the combination of both agents provided the benefits of prompt relief afforded by BW942C and lasting relief afforded by TMP-SMX. BW942C might prove to be an agent suitable for the treatment of acute diarrhea, with TMP-SMX reserved for treatment of those who do not respond adequately. The empiric use of the combination of BW942C and TMP-SMX appears appropriate for the treatment of severe nondysenteric disease.
Selective antimicrobial modulation of the intestinal tract by norfloxacin in human volunteers and in gnotobiotic mice associated with a human fecal flora
Authors: Pecquet S , Andremont A , Tancrède C
Published: June 2012

Intestinal endogenous members of the family Enterobacteriaceae were eliminated in 12 human volunteers treated with 400 or 800 mg of oral norfloxacin per day for 5 days. No clones resistant to quinolone derivatives were isolated. Counts of aerotolerant streptococci were affected to various degrees, depending on their susceptibility to norfloxacin. During treatment, counts of anaerobes remained above 9.8 log10 CFU/g of feces. A total of 932 anaerobic isolates from the predominant flora (over 10(9) CFU/g) in fecal samples obtained before or during norfloxacin treatment were classified by a simple morphological and physiological scheme. The composition of this flora was fairly stable from one sample to another before treatment and was not substantially modified by norfloxacin. Intestinal resistance to colonization by exogenous microorganisms was studied in gnotobiotic mice associated with a human fecal flora. The composition of the fecal flora of the human donor and the fecal concentrations of norfloxacin in the volunteers were reproduced in the intestine of the mice. Resistance to colonization by exogenous microorganisms was reduced by norfloxacin for only 2 of 14 (14%) of the strains tested. These results suggest that norfloxacin is a good candidate for selective antimicrobial modulation of the intestinal tract in humans.
In vitro activity of Ro 15-8074 and Ro 19-5247, two orally administered cephalosporin metabolites
Authors: Wise R , Andrews J M , Piddock L J
Published: June 2012

The activity of two iminomethoxy aminothiazoly cephalosporins, Ro 15-8074 and Ro 19-5247, was compared with that of other beta-lactams against a total of 491 bacterial strains. Both were highly active (MIC for 90% of the strains tested [MIC 90], less than or equal to 2 micrograms/ml) against the majority of the members of the family Enterobacteriaceae, Haemophilus influenzae, Neisseria spp., and Streptococcus pneumoniae, being at least 16-fold more active than cephalexin and 8-fold more active than cefuroxime. There was no activity against Pseudomonas aeruginosa and poor activity against Morganella morganii (in the case of Ro 15-8074), Enterobacter sp., and Citrobacter sp. Staphylococcus aureus was moderately susceptible to Ro 19-5247 (MIC90, 8 micrograms/ml), but Ro 15-8074 was eightfold less active. The protein binding of the two compounds at 5 micrograms/ml was 9.1% for Ro 15-8074 and 69.9% for Ro 19-5247. The major target site for the two cephalosporins was PBP 3.
Effect of 4-quinolones and novobiocin on calf thymus DNA polymerase alpha primase complex, topoisomerases I and II, and growth of mammalian lymphoblasts
Authors: Hussy P , Maass G , Tümmler B , Grosse F , Schomburg U
Published: June 2012

The influence of ciprofloxacin, nalidixic acid, norfloxacin, novobiocin, and ofloxacin on elements of eucaryotic DNA replication was investigated in vitro. Each of the 4-quinolones, when present in amounts of more than 100 micrograms/ml, reversibly inhibited the DNA synthesis performed by the 95 DNA polymerase alpha primase complex from calf thymus. Novobiocin at 500 micrograms/ml or at higher concentrations irreversibly inactivated DNA polymerase alpha primase complex. The accuracy of in vitro DNA synthesis in the absence of repair mechanisms was determined from amber-revertant assays with phi X174am16(+) DNA as template. The antimicrobial agents did not significantly increase the frequencies of base pairing mismatches during the course of replication, indicating that the basal mutation rate is not affected by novobiocin and the 4-quinolones. The Ki values of 50% inhibition of DNA topoisomerases from calf thymus by ciprofloxacin, norfloxacin, novobiocin, nalidixic acid, and ofloxacin were 300, 400, 1,000 or more, 1,000 or more, and 1,500 or more micrograms/ml, respectively, in the case of topoisomerase I, and the Ki values were 150, 300, 500, 1,000, and 1,300 micrograms/ml, respectively, in the case of topoisomerase II. The procaryotic topoisomerase II is approximately 100-fold more sensitive to inhibition by ciprofloxacin, norfloxacin, and ofloxacin than is its eucaryotic counterpart. Growth curves of lymphoblasts were recorded in the presence of ofloxacin and ciprofloxacin. Neither 1 nor 10 micrograms of ciprofloxacin or of ofloxacin per ml affected cell proliferation. Ofloxacin and ciprofloxacin at 100 micrograms/ml inhibited cell growth; 1,000 micrograms/ml led to cell death. No correlation exists between the antimicrobial and cytotoxic activities of the 4-quinolones.
Comparative activities of LY 164846 and other orally administered beta-lactam antibiotics against Haemophilus influenzae
Authors: Jorgensen J H , Redding J S , Howell A W
Published: June 2012

LY 164846, a new oral cephalosporin, demonstrated very good inhibitory and bactericidal activities against Haemophilus influenzae irrespective of the production of beta-lactamase by the test strains. However, its activity was not markedly superior to that of either Augmentin (amoxicillin plus clavulanate) or cefaclor.
Epidemiology of intestinal colonization by members of the family Enterobacteriaceae highly resistant to erythromycin in a hematology-oncology unit
Authors: Andremont A , Sancho-Garnier H , Tancrede C
Published: June 2012

Intestinal colonization by highly erythromycin-resistant members of the family Enterobacteriaceae was surveyed for 4 years in a hematology-oncology unit. Fifty-four of 555 patients (9.7%) were colonized, each with a different strain. The incidence of intestinal carriage was not correlated with erythromycin consumption in the ward but was strongly associated with individual exposure to erythromycin.
Cephalosporin-induced hypoprothrombinemia: is the N-methylthiotetrazole side chain the culprit?
Authors: Agnelli G , Del Favero A , Parise P , Guerciolini R , Pasticci B , Nenci G G , Ofosu F
Published: June 2012

The reported high incidence of vitamin-K-reversible hypoprothrombinemia associated with the new beta-lactamase-stable cephalosporins prompted us to evaluate the effect on hemostasis of three cephalosporins (cefamandole, ceftriaxone, and ceftazidime) in 30 patients with serious infections. Cefamandole and ceftriaxone, both containing a sulfhydryl group, induced a significant and similar prolongation of prothrombin time and decrease in factor VII activity. Ceftazidime, in contrast, had no effect on these two parameters.
In vitro activity of BRL 36650, a new semisynthetic penicillin
Authors: Hoy J F , Rolston K V , Ho D H , Alvarez M , Thirolf P , Bodey G P
Published: June 2012

BRL 36650 [sodium 6 beta-(D-2-[(4-ethyl-2, 3-dioxopiperazin-1-yl)carbonylamino]-2-(3,4-dihydroxyphenyl) acetamido)-6 alpha-formamido-penicillinate] is a new semisynthetic penicillin. It was tested in vitro for activity against 884 organisms cultured from blood specimens of cancer patients. BRL 36650 had broad-spectrum activity against the gram-negative bacilli tested but had no gram-positive activity. The MIC against 90% of the Pseudomonas aeruginosa isolates was 3.12 micrograms/ml. The activity of BRL 36650 was superior to that of piperacillin, comparable or slightly inferior to that of aztreonam and ceftazidime, and lower than that of imipenem and amifloxacin. BRL 36650 should prove useful for the management of gram-negative bacillary infections, including those caused by P. aeruginosa.
Column-switching high-performance liquid chromatographic method for determination of a new antiviral agent, cyclaradine, in serum
Authors: Lim J M , Lin C C
Published: June 2012

Cyclaradine is a novel carbocyclic nucleoside with good activity against the viruses of the herpes group. To facilitate pharmacokinetic studies on cyclaradine, an automated column-switching high-performance liquid chromatographic (HPLC) method was developed for the determination of cyclaradine in serum. Deproteinized serum containing cyclaradine was directed to an on-line extraction column which retained cyclaradine while many potentially interfering components were eluted to waste. Fourteen minutes later, the switching valve was automatically rotated, permitting an appropriate mobile phase to elute cyclaradine from the extraction column onto an analytical C18 column for further separation and UV detection. The method showed excellent linearity (r = 0.9995) for cyclaradine concentrations ranging from 0.05 to 5 micrograms/ml. It also provided good sensitivity (0.05 micrograms/ml). The assay was precise, with within-run and between-run coefficients of variation of less than 1.90%. The accuracy, expressed as differences between observed values and theoretical values, ranged from -4.12 to 4.80%. The assay involves a simple deproteinization of serum followed by a fully automated sample cleanup, eliminating long and tedious manual extraction prior to HPLC. The column-switching HPLC method has been successfully used for the determination of cyclaradine serum levels in squirrel monkeys following a single oral dose of 20 mg/kg.
Multiple low-level antibiotic resistance in Aeromonas salmonicida
Authors: Wood S C , McCashion R N , Lynch W H
Published: June 2012

Mutants with multiple low-level antibiotic resistance were isolated from virulent wild-type Aeromonas salmonicida strains exposed to a low concentration of any one of several low-molecular-mass (approximately 635 daltons or less) antibiotics. Multiple resistance was toward beta-lactam compounds (penicillin G, ampicillin, cloxacillin), quinolones (flumequine, oxolinic acid, nalidixic acid), tetracyclines, chloramphenicol, and novobiocin. Susceptibilities of the mutants toward several higher-molecular-mass (greater than 700 daltons) hydrophobic or polycationic antibiotics such as rifampin, erythromycin, polymyxin B, and streptomycin sulfate were not affected. The mutants were obtained at frequencies suggesting point mutations. Outer membrane protein profiles, examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed that all multiple low-level resistant mutants were deficient in a major protein of approximately 38.5 kilodaltons and contained a major protein of approximately 37 kilodaltons which was not present in significant amounts in the wild-type strains. In addition, these mutants lacked exoprotease activity. Furthermore, mutants isolated as deficient in exoprotease were found, with the exception of one avirulent strain, to exhibit multiple low-level antibiotic resistance and the outer membrane protein changes.
Occurrence of clindamycin-resistant anaerobic bacteria isolated from cultures taken following clindamycin therapy
Authors: Ohm-Smith M J , Sweet R L , Hadley W K
Published: June 2012

MICs of clindamycin were determined by the agar dilution method against anaerobic organisms isolated from endometrial cultures in women with pelvic soft tissue infections. Cultures were obtained from 100 women both before and after clindamycin therapy, from 107 women before therapy with clindamycin or another antimicrobial agent or after treatment with an antimicrobial agent other than clindamycin, and from 9 women 1 to 9 weeks after they were discharged from the hospital following clindamycin therapy. Only 5 (0.7%) of 685 isolates tested from women who had not received clindamycin therapy were resistant to clindamycin. From the 100 cultures taken immediately after clindamycin therapy, 57 anaerobic bacteria were isolated from 28 cultures. Of the 40 anaerobic organisms for which MICs of clindamycin were determined, 25 (62.5%) were resistant to clindamycin (MIC greater than or equal to 8 micrograms/ml). The most common organisms isolated after therapy were the anaerobic gram-positive cocci (of which 32 isolates were discovered); of 28 coccal isolates tested, 64% were clindamycin resistant. Four of seven (57%) of the Bacteroides isolates tested, one unidentified gram-positive nonsporing rod, one unidentified gram-negative coccus, and one Mobiluncus sp. were also clindamycin resistant. Of 18 anaerobic isolates from the nine cultures taken 1 to 9 weeks after hospital discharge, 55% were resistant to clindamycin. The clinical significance of these findings is unknown since all patients recovered without incident and remained well. However, the data suggest that physicians need to be aware that patients with recent exposure to clindamycin may have clindamycin-resistant anaerobic organisms in a current infection. This may prevent the infection from responding to clindamycin treatment.
Synergy between RU 28965 (roxithromycin) and human neutrophils for bactericidal activity in vitro
Authors: Labro M T , Amit N , Babin-Chevaye C , Hakim J
Published: June 2012

The in vitro effects of RU 28965 (roxithromycin), a new semisynthetic macrolide, on human neutrophil activity were compared with those of erythromycin. RU 28965, at a concentration as low as 0.1 microgram/ml, significantly enhanced the phagocytosis and killing of Staphylococcus aureus by neutrophils. Erythromycin displayed a less stimulating effect in a dose-dependent manner. Phagocytosis of Klebsiella pneumoniae was also increased after incubation of neutrophils with RU 28965, but killing was not altered. Neutrophil chemotaxis, myeloperoxidase activity, and O2 consumption were unchanged in the presence of RU 28965.
Lack of antibacterial activity of pentoxifylline
Authors: Yu P K , Washington J A
Published: June 2012

Pentoxifylline was inactive when tested against 70 fresh clinical bacterial isolates, as well as when tested alone and in combination with erythromycin, vancomycin, or rifampin against Escherichia coli PC1349 (wild type) and its heptose-deficient outer membrane mutant E. coli PC2040. No antibacterial activity of pentoxifylline or its metabolites could be detected in sera from patients receiving pentoxifylline or its metabolites could be detected in sera from patients receiving pentoxifylline.
Potentiation by levamisole, methisoprinol, and adenine or adenosine of the inhibitory activity of human interferon against encephalomyocarditis virus
Authors: Muñoz A , García R A , Pérez-Aranda A
Published: June 2012

The antiviral action of different human interferons against encephalomyocarditis virus in HeLa cell cultures was analyzed. Cell treatment with levamisole (200 micrograms/ml), methisoprinol (1 mg/ml), or adenine or adenosine (33 or 100 micrograms/ml, respectively) potentiated the anti-encephalomyocarditis virus activity of human interferon by 8- to 32-fold. A higher level of potentiation (256-fold) was achieved either by combined treatments with levamisole plus methisoprinol or by treatment with one of these compounds plus adenine or adenosine.
Comparison of foscarnet cream, acyclovir cream, and acyclovir ointment in the topical treatment of experimental cutaneous herpes simplex virus type 1 infection
Authors: Spruance S L , Freeman D J , Sheth N V
Published: June 2012

Topical foscarnet (PFA) and acyclovir (ACV) were compared in the dorsal cutaneous guinea pig model of herpes simplex virus type 1 infection. The relative order of efficacy was PFA cream greater than ACV cream greater than ACV ointment. In vitro studies demonstrated that PFA and ACV formulated in cream vehicles penetrated through guinea pig skin 7- to 10-fold faster than did ACV in ointment.