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1.  Novel gyrA point mutation in a strain of Escherichia coli resistant to fluoroquinolones but not to nalidixic acid. 
We have previously described a clinical isolate of Escherichia coli (Q2) that is highly resistant to fluoroquinolones (MIC of ciprofloxacin, 16 micrograms/ml) but susceptible to nalidixic acid (MIC of nalidixic acid, 4 micrograms/ml) (N. Moniot-Ville, J. Guibert, N. Moreau, J.F. Acar, E. Collatz, and L. Gutmann, Antimicrob. Agents Chemother. 35:519-523, 1991). Transformation of strain Q2 with a plasmid carrying the wild-type gyrA gene from E. coli K-12(pAFF801) resulted in a 32-fold decrease in the MIC of ciprofloxacin, suggesting that at least one mutation in gyrA was involved in the resistance of Q2. Intragenic gyrA fragments of 668 and 2,500 bp from strain Q2 were amplified by the polymerase chain reaction. We sequenced the 668-bp fragment and identified a single novel point mutation (transition from G to A at position 242), leading to an amino acid substitution (Gly-81 to Asp) in the gyrase A subunit. We constructed hybrid plasmids by substituting either the 668-bp fragment or the 2,500-bp fragment from Q2 DNA, both of which contained the gyrA point mutation, for the corresponding fragments in wild-type gyrA (2,625 bp) of E. coli K-12. When introduced into E. coli KNK453 (gyrA temperature sensitive), both plasmids conferred an eightfold increase in the MIC of ciprofloxacin, but only a twofold increase in the MIC of nalidixic acid. When introduced into E. coli Q2, neither plasmid conferred any change in the MICs of ciprofloxacin or nalidixic acid, suggesting that only the point mutation found in gyrA was involved in the resistance that we observed.
PMCID: PMC187948  PMID: 8392306
2.  Activity of ampicillin-sulbactam and oxacillin in experimental endocarditis caused by beta-lactamase-hyperproducing Staphylococcus aureus. 
Using a rat model of aortic valve infective endocarditis, we previously found that oxacillin was equally effective against an oxacillin-susceptible strain of Staphylococcus aureus and a beta-lactamase-hyperproducing borderline oxacillin-susceptible strain of S. aureus; also, ampicillin-sulbactam was less effective than oxacillin against both isolates and at low doses was less effective against the borderline-susceptible strain than against the fully oxacillin-susceptible strain (C. Thauvin-Eliopoulos, L. B. Rice, G. M. Eliopoulos, and R. C. Moellering, Jr., Antimicrob. Agents Chemother. 34:728-732, 1990). In the present study, we extended this work, using alternative treatment schedules and additional bacterial strains. Extending treatment with low doses of ampicillin-sulbactam (500 and 250 mg/kg of body weight per day, respectively) to 6.5 days resulted in equalization of effectiveness against the previously studied strains BOSSA-1 and OSSA-1 (3.75 +/- 1.61 log10 and 4.71 +/- 1.79 log10 CFU of residual viable bacteria per g, respectively). Against the borderline oxacillin-susceptible strain BOSSA-1, increasing the sulbactam dosage from 500 to 2,000 mg/kg/day while maintaining a fixed dose of ampicillin (1,000 mg/kg/day) by continuous infusion resulted in lower bacterial counts (4.93 +/- 1.84 log10 versus 3.65 +/- 1.26 log10 CFU of residual viable bacteria per g, respectively), but this difference was of only borderline significance; differences in efficacy between the low-dose and high-dose sulbactam regimens were exaggerated when intermittent intravenous administration was used (6.19 +/- 1.90 log10 versus 3.37 +/- 1.41 log10 CFU/g, respectively; P < 0.001). However, for any individual sulbactam dosage, the model of administration (continuous versus intermittent infusion) did not affect the activity of the regimen. When additional strains were used in the model, oxacillin and ampicillin-sulbactam (1,000 plus 2,000 mg/kg/day) were equally effective against both oxacillin-susceptible and borderline oxacillin-resistant strains of S. aureus. These results support the predictions that oxacillin would be clinically effective in the treatment of infections caused by borderline oxacillin-susceptible strains of S. aureus and that, except at very low doses, ampicillin-sulbactam would also be as effective against borderline-susceptible strains as against fully oxacillin-susceptible strains of S. aureus.
PMCID: PMC187700  PMID: 8460919
3.  In vitro activities of ceftriaxone and fusidic acid against 13 isolates of Coxiella burnetii, determined using the shell vial assay. 
The susceptibilities of 13 isolates of Coxiella burnetii to fusidic acid and ceftriaxone were determined by use of the recently described shell vial assay (D. Raoult, H. Torres, and M. Drancourt, Antimicrob. Agents Chemother, 35:2070-2077, 1991). At a concentration of 4 micrograms/ml, ceftriaxone was bacteriostatic for four isolates and slowed the multiplication of the other nine. Fusidic acid at a concentration of 2 micrograms/ml was bacteriostatic for six isolates and slowed the multiplication of three others. These results show that these compounds could be effective in the phagolysosome of C. burnetii-infected cells.
PMCID: PMC187697  PMID: 8460917
4.  Efficacy of intravenous itraconazole against experimental pulmonary aspergillosis. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2762-2765.
The efficacy of intravenous itraconazole solubilized in hydroxypropyl-beta-cyclodextrin was assessed in a rat model of Aspergillus fumigatus pneumonia. Immunosuppressed rats were infected by intratracheal inoculation of A. fumigatus conidia. Intravenous administration of various doses of itraconazole was started immediately after infection and continued once a day for 7 days. A 10-mg dose of intravenous itraconazole per kg was as effective on survival as 1 mg of amphotericin B per kg daily (a survival rate of 100% in 28 days), while treatment with 1 mg/kg did not increase the survival rate. The 50% lethal dose of intravenous itraconazole given to immunosuppressed and uninfected rats for 7 days was 24.5 mg/kg/day. A microbiological assay to estimate accumulation in tissue after five daily intravenous administrations of itraconazole at 10 mg/kg showed that itraconazole and its active metabolites were present in the lungs for at least 6 h, reaching the MIC as previously described (B. Dupont and E. Drouchet, Rev. Infect. Dis. 9(Suppl. 1):71-76, 1987; A. Espinel-Ingroff, S. Shadomy, and R. J. Gebhart, Antimicrob. Agents Chemother. 26:5-9, 1984). Intravenous itraconazole was considered to be worth evaluating in clinical trials of aspergillosis.
PMCID: PMC192807  PMID: 8109952
5.  Analysis of the aac(3)-VIa gene encoding a novel 3-N-acetyltransferase. 
Antimicrobial Agents and Chemotherapy  1993;37(10):2074-2079.
Biochemical analysis (G. A. Papanicolaou, R. S. Hare, R. Mierzwa, and G. H. Miller, abstr. 152, Program Abstr. 29th Intersci. Conf. Antimicrob. Agents Chemother., 1989) demonstrated the presence of a novel 3-N-acetyltransferase in Enterobacter cloacae 88020217. This organism was resistant to gentamicin, and the MIC of 2'-N-ethylnetilmicin for it was fourfold lower than that of 6'-N-ethylnetilmicin, a resistance pattern which suggested 2'-acetylating activity. However, high-pressure liquid chromatography analysis demonstrated that the enzyme acetylated sisomicin in the 3 position. We have cloned the structural gene for this enzyme from a large (> 70-kb) conjugative plasmid present in E. cloacae. Subcloning experiments have localized the aac(3)-VIa gene to a 2.1-kb Sau3A fragment. The deduced AAC(3)-VIa protein showed 48% amino acid identity to the AAC(3)-IIa protein and 39% identity to the AAC(3)-VII protein. Examination of the 5'-flanking sequences demonstrated that the aac(3)-VIa gene was located 167 bp downstream of the aadA1 gene and was present in an integron. In addition, the aac(3)-VIa gene is also downstream of a 59-base element often seen in an integron environment. Primer extension analysis has identified a promoter for the aac(3)-VIa gene downstream of both the aadA1 gene and a 59-base element.
PMCID: PMC192231  PMID: 8257126
6.  8-aminoquinolines effective against Pneumocystis carinii in vitro and in vivo. 
Antimicrobial Agents and Chemotherapy  1993;37(10):2166-2172.
The activities of 25 8-aminoquinolines were compared in tests assessing the ability of the compounds to inhibit the growth of Pneumocystis carinii in culture. Six compounds were effective at or below 0.03 microM: CDRI 80/53, NSC19894, NSC305805, NSC305812, WR182234, and primaquine. Four others were effective at between 0.2 and 0.03 microM: NSC305835, WR225448, WR238605, and WR242511. Fourteen drugs were also tested in a standard model of P. carinii pneumonia in rats at daily doses of 2 mg/kg of body weight in drinking water. CDRI 80/53, NSC305805, NSC305835, and WR225448 were extremely effective in the animal model. The effectiveness of WR238605, WR242511, and primaquine in the rat model has been reported elsewhere (M. S. Bartlett, S. F. Queener, R. R. Tidwell, W. K. Milhouse, J. D. Berman, W. Y. Ellis, and J. W. Smith, Antimicrob. Agents Chemother. 35:277-282, 1991). The length of the alkyl chain separating the nitrogens in the substituent at position 8 of the quinoline ring was a strong determinant of anti-P. carinii activity.
PMCID: PMC192245  PMID: 8257140
7.  Interaction of gentamicin with the A band and B band lipopolysaccharides of Pseudomonas aeruginosa and its possible lethal effect. 
The lipopolysaccharide (LPS) of Pseudomonas aeruginosa PAO1 possesses two distinct types of O polysaccharide, A and B band LPSs, but the majority of clinical isolates from cystic fibrosis patients who are infected with the organism possess only the A band as the major LPS antigen. The initial step in a series of events during the uptake of aminoglycoside antibiotics such as gentamicin is the ionic binding of the molecule to the cell surface. In an attempt to elucidate the role of A and B band LPSs of P. aeruginosa in this passive ionic binding of gentamicin to the outer membrane and its possible lethal effects, strains PAO1 (A+B+) and LPS isogenic derivatives (A+B-,A-B+,A-B-) were treated with the antibiotic. Ionic binding of gentamicin appeared to be subtly different in PAO1 and its LPS derivatives; a lethal dose of drug was bound to all strains, although the degree of binding varied with each strain. The outer membrane affinity for gentamicin was higher in strains possessing the B band than in strains with A band LPS, and these B band strains were more prone to antibiotic-induced killing. Strains with both A and B band LPSs bound the most gentamicin of all strains, and this binding caused an almost 50% loss in viability. Ionic binding of aminoglycoside antibiotucs to the outer membrane of cell surfaces must not only weaken th cell surface (R. E. W. Hancock, Annu. Rev. Microbiol. 38:237-264, 1984; N. L. Martin and T. J. Beveridge, Antimicrob. Agents Chemother. 29:1079-1087, 1986; S. G. Walker and T. J. Beveridge, Can. J. Microbiol. 34:12-18, 1988) but it must also be more important in cell death than was originally thought.
PMCID: PMC187740  PMID: 8494366
8.  Nucleotide sequence analysis and DNA hybridization studies of the ant(4')-IIa gene from Pseudomonas aeruginosa. 
The ant(4')-IIa gene was previously cloned from Pseudomonas aeruginosa on a 1.6-kb DNA fragment (G. A. Jacoby, M. J. Blaser, P. Santanam, H. Hächler, F. H. Kayser, R. S. Hare, and G. H. Miller, Antimicrob. Agents Chemother. 34:2381-2386, 1990). In the current study, the ant(4')-IIa gene was localized by gamma-delta mutagenesis. A region of approximately 600 nucleotides which contained the ant(4')-IIa gene was identified, and DNA sequence analysis revealed two overlapping open reading frames (ORFs) within this region. Northern (RNA) blot analysis demonstrated expression of both ORFs in P. aeruginosa; therefore, site-directed mutagenesis was used to identify the ORF which encodes the ant(4')-IIa gene. No homology was found between ant(4')-IIa and ant(4')-Ia DNA sequences. Hybridization experiments confirmed that the ant(4')-Ia probe hybridized only to gram-positive presumptive ANT(4')-I strains and that the ant(4')-IIa probe hybridized only to gram-negative strains presumed to carry ANT(4')-II. Seven gram-negative strains which had been classified as having ANT(4')-II resistance profiles did not hybridize with probes for either ant(4')-Ia or ant(4')-IIa, suggesting that at least one additional ant(4') gene may exist. The predicted amino-terminal sequences of the ANT(4')-Ia and ANT(4')-IIa proteins showed significant sequence similarity between residues 38 and 63 of the ANT(4')-Ia protein and residues 26 and 51 of the ANT(4')-IIa protein.
PMCID: PMC187739  PMID: 8494365
9.  Mutations in the gyrA gene of a highly fluoroquinolone-resistant clinical isolate of Escherichia coli. 
We have determined the DNA sequence of the gyrA gene of the fluoroquinolone-resistant Escherichia coli isolate 205096 (MIC of ciprofloxacin, 128 micrograms/ml), which was recently demonstrated to be a gyrA mutant (P. Heisig and B. Wiedemann, Antimicrob. Agents Chemother. 35:2031-2036, 1991). Compared with the gyrA+ gene of E. coli K-12, 55 nucleotide changes were found. Three of these resulted in amino acid exchanges: Ser-83-->Leu, Asp-87-->Gly, and Asp-678-->Glu. A 0.7-kb DNA fragment containing two of these mutations (Ser-83-->Leu and Asp-87-->Gly) was isolated and fused in frame to the residual 3' coding region of gyrA+ in a plasmid to yield a chimeric gyrA gene (gyrA#). After introduction into E. coli 205096, this gyrA# gene does not increase the fluoroquinolone susceptibility of the resulting heterodiploid strain in a dominance test, while the gyrA+ gene does. The ciprofloxacin concentration necessary to inhibit by 90% (IC90) the supercoiling activity of gyrase isolated from E. coli 205096 is above 2,000 micrograms/ml. An identical result was found for gyrase reconstituted in vitro from the gyrB+ gene product and the chimeric gyrA# gene product. This is more than a 4,000-fold increase compared with the IC90 determined for gyrase from E. coli K-12 (gyrA+) (IC90, 0.5 microgram of ciprofloxacin per ml). No indications for the involvement of the gyrB gene or for alterations in quinolone permeation were found.
PMCID: PMC187737  PMID: 8388197
10.  Mechanisms of action of cephalosporin 3'-quinolone esters, carbamates, and tertiary amines in Escherichia coli. 
Cephalosporin 3'-quinolone esters, carbamates, and tertiary amines are potent antibiotics whose antibacterial activities reflect the action of both the beta-lactam and the quinolone components. The biological properties of representative compounds from each class were compared in Escherichia coli. All compounds bound to the essential PBP 3, inhibited DNA gyrase, and caused filamentation in growing cells. To distinguish between cephalosporin- and quinolone-induced filaments, nucleoid segregation was also examined, as quinolones disrupt nucleoid segregation while the beta-lactams do not (N. H. Georgopapadakou and A. Bertasso, Antimicrob. Agents Chemother. 35:2645-2648, 1991). The cephalosporin quinolone esters Ro 23-9424 and Ro 24-6392, at concentrations causing filamentation in E. coli ATCC 25922, did not affect nucleoid segregation after 1 h of incubation (cephalosporin response) but did not affect it after 2 h (quinolone response), indicating the release of free quinolone. Accordingly, only the quinolone response was produced in a strain possessing TEM-3, an expanded-spectrum beta-lactamase. The cephalosporin carbamate Ro 24-4383 and the tertiary amine Ro 24-8138 produced a quinolone response in E. coli ATCC 25922, though they produced a cephalosporin response in a quinolone-resistant strain. Carbamate and tertiary amine linkages are chemically more stable than the ester linkage, and both cephalosporin 3'-quinolone carbamates and tertiary amines are more potent inhibitors of DNA gyrase than are the corresponding esters. The results suggest that, while intact cephalosporin 3'-quinolone esters act as cephalosporins, carbamates and amines may possess both cephalosporin and quinolone activity in the intact molecule.
PMCID: PMC187707  PMID: 8384817
17.  In vitro activity of DU-6859a, a new fluorocyclopropyl quinolone. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2747-2753.
DU-6859a was tested against 844 recent clinical isolates (most from bacteremias) by using reference MIC determination procedures. The activity of DU-6859a against members of the family Enterobacteriaceae was comparable to that of ciprofloxacin (range of MICs for 90% of isolates [MIC90], < or = 0.015 to 1 microgram/ml), and the highest MICs were observed among Serratia marcescens and Providencia rettgeri isolates. The DU-6859a MIC90 for Pseudomonas aeruginosa and Xanthomonas maltophilia was 0.5 microgram/ml. Pneumococci (MIC90, 0.06 microgram/ml), Haemophilus influenzae (MIC90, < or = 0.004 microgram/ml), Moraxella catarrhalis (MIC90, < or = 0.015 microgram/ml), and pathogenic neisseriae (MIC90, 0.015 to 0.03 microgram/ml) were very susceptible to DU-6859a. All staphylococci had DU-6859a MICs of < or = 1 microgram/ml, including oxacillin- and ciprofloxacin-resistant strains. DU-6859a was very active against isolates resistant to ceftazidime (MIC90, < or = 0.12 microgram/ml), ciprofloxacin (MIC90, < or = 8 micrograms/ml), and gentamicin (MIC90, < or = 1 microgram/ml).
PMCID: PMC192801  PMID: 8109948
18.  Transient carbapenem resistance induced by salicylate in Pseudomonas aeruginosa associated with suppression of outer membrane protein D2 synthesis. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2743-2746.
Pseudomonas aeruginosa PAO1 showed increased phenotypic resistance to imipenem, panipenem, and biapenem specifically in the presence of salicylate. The antipseudomonal activity of carbapenems was reduced in proportion to the concentration of salicylate. This resistance was transient and nonheritable. The synthesis of the outer membrane protein D2 (OprD or OprD2) in P. aeruginosa PAO1 was inhibited by 4 to 32 mM salicylate in the bacterial growth medium, whereas no changes in any other outer membrane proteins were observed. These results indicate that salicylate suppresses the synthesis of OprD and therefore reduces the antipseudomonal activity of carbapenems. Under these conditions, one carbapenem--meropenem--is still active against P. aeruginosa, which indicates that meropenem can pass through the outer membrane via both the D2 channel and another undefined route(s).
PMCID: PMC192800  PMID: 8109947
19.  Antimicrobial susceptibilities of Gardnerella vaginalis. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2733-2735.
The in vitro susceptibilities of 93 clinical isolates of Gardnerella vaginalis to 25 antimicrobial agents were determined by the agar dilution method. All isolates were susceptible to penicillin, ampicillin, erythromycin, clindamycin, chloramphenicol, and trimethoprim. Activity was poor for vancomycin, LY146032, the cephalosporins, ciprofloxacin, and imipenem. Some resistance was observed with tetracycline and minocycline. The MICs of metronidazole paralleled those of tinidazole, with the hydroxymetabolite of metronidazole being the most active. One strain was resistant to all three agents. Marked resistance to aztreonam, amikacin, and sulfamethoxazole was observed.
PMCID: PMC192794  PMID: 8109944
20.  A new tetracycline resistance determinant, Tet H, from Pasteurella multocida specifying active efflux of tetracycline. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2699-2705.
The tetracycline resistance determinant on plasmid pVM111 from an avian strain of Pasteurella multocida mediates tetracycline resistance by a regulated active efflux mechanism. DNA coding for the determinant did not hybridize at high stringency with DNA representing a group of common tetracycline resistance determinants. The DNA sequence, however; revealed a structural gene and a repressor gene which had significant (37 to 64%) sequence similarities with previously described classes of efflux-type tetracycline resistance genes from members of the family Enterobacteriaceae. The new determinant has been assigned to class H.
PMCID: PMC192782  PMID: 8109938
21.  Regional and systemic prophylaxis with teicoplanin in monolateral and bilateral total knee replacement procedures: study of pharmacokinetics and tissue penetration. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2693-2698.
Twenty-four patients undergoing monolateral or bilateral total knee replacement (TKR) procedures were randomized to receive teicoplanin (T) either systemically or regionally. Subjects scheduled for systemic prophylaxis and undergoing monolateral (six patients) or bilateral (five patients) TKR received a single 800-mg dose of T in 100 ml of saline as a 5-min infusion into a forearm vein 2.5 h before surgery. For regional prophylaxis, patients undergoing monolateral surgery (eight subjects) received 400 mg of T in 100 ml of saline as a 5-min infusion into a foot vein of the leg to be operated on immediately after the tourniquet was inflated. For the five patients scheduled for bilateral operation and regional prophylaxis, the administration of T was also repeated for the second knee operation. The tourniquet, as the standard TKR surgical technique, was inflated to 400 mm Hg (c. 50 kPa) in all 24 patients immediately before the beginning of surgery and kept in place for the duration of the operation. Samples of serum, bone, skin, synovia, and subcutaneous tissue were collected at timed intervals during surgery. They were microbiologically assayed for T by using Bacillus subtilis as the test organism. Overall, the mean T concentrations obtained with regional route prophylaxis were found to be 2 to 10 times higher than those achieved following systemic prophylaxis. Moreover, peak levels in different tissues after regional prophylaxis were significantly higher (P < 0.05). None of the patients experienced adverse effects due to regional or systemic T administration; no prosthetic or wound infections were observed in the follow-up period (from 12 to 26 months).
PMCID: PMC192781  PMID: 8109937
22.  In vitro antibacterial activities of PD 138312 and PD 140248, new fluoronaphthyridines with outstanding gram-positive potency. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2563-2570.
PD 138312 and PD 140248 are new quinolones with high in vitro activities against a wide spectrum of bacterial species, notably including gram-positive isolates. The respective MICs (in micrograms per milliliter) of PD 138312 and PD 140248 capable of inhibiting > or = 90% of the strains were < or = 0.06 and < or = 0.06 for oxacillin-susceptible and -resistant staphylococci, streptococci (including Streptococcus pyogenes, S. agalactiae, S. pneumoniae, and viridans group streptococci), Haemophilus influenzae, Moraxella catarrhalis, and Neisseria gonorrhoeae; 0.125 and 0.03 for Legionella pneumophila; 0.25 and 0.125 for Listeria monocytogenes; 0.25 and 0.25 for Enterococcus faecalis; 0.5 and 0.06 for anaerobic gram-positive cocci; 0.5 and 0.25 for Acinetobacter spp.; 0.5 and 0.5 for members of the family Enterobacteriaceae (excluding Serratia marcescens); 2 and 0.5 for Bacteroides fragilis; 2 and 2 for Serratia marcescens and ciprofloxacin-resistant staphylococci; and 8 and 4 for Pseudomonas aeruginosa.
PMCID: PMC192739  PMID: 8109918
23.  Biliary excretion of rufloxacin in humans. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2545-2549.
Rufloxacin is a new once-daily antibacterial fluoroquinolone with a long half-life. The aim of the present study was to evaluate the plasma and biliary kinetics and biliary and urinary excretion of rufloxacin in patients with extrahepatic cholestasis. Twelve patients with total external percutaneous transhepatic biliary drainage were given a single oral dose of 400 mg of rufloxacin. Plasma, bile, and urine samples and fractions were collected over 72 h after drug administration. Rufloxacin and its major metabolite, the N-desmethyl derivative, were measured by high-performance liquid chromatography. Maximum rufloxacin concentrations in plasma and bile (means +/- standard deviations) were 4.05 +/- 1.38 micrograms/ml and 8.24 +/- 7.16 micrograms/ml, respectively, and were reached in 4.2 +/- 3.0 h and 4.2 +/- 3.5 h, respectively. The terminal elimination half-life of rufloxacin in plasma was 45.1 +/- 13.5 h. Apparent plasma clearance was 31.3 +/- 10.5 ml/min, while biliary clearance was 0.4 +/- 0.2 ml/min and renal clearance was 12.7 +/- 6.0 ml/min. In 72 h, 0.9% +/- 0.8% of the dose given was recovered in bile and 27.2% +/- 12.0% was recovered in urine. Biliary concentrations exceeded the MICs of most common biliary tract pathogens for at least 24 h after administration. The broad antibacterial spectrum of rufloxacin and its high and prolonged biliary concentrations suggest that this drug may be useful for treatment of biliary tract infections.
PMCID: PMC192734  PMID: 8109915
24.  Study of heterogeneity of chloramphenicol acetyltransferase (CAT) genes in streptococci and enterococci by polymerase chain reaction: characterization of a new CAT determinant. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2593-2598.
An assay based on the utilization of degenerate primers that enable enzymatic amplification of an internal fragment of cat genes known to be present in gram-positive cocci was developed to identify the genes encoding chloramphenicol resistance in streptococci and enterococci. The functionality of this system was illustrated by the detection of cat genes belonging to four different hydridization classes represented by the staphylococcal genes catpC221, catpC194, catpSCS7, and the clostridial gene catP, and by the characterization of a new streptococcal cat gene designated catS. A sequence related to the clostridial catQ gene, which was present in one streptococcal strain, was not detected by this assay. These results reveal that these six cat genes account for chromosomal-borne chloramphenicol resistance in 12 group A, B, and G streptococci tested. By contrast, only three of these six cat genes (catpC221, catpC194, and catpSCS7) were detected on the 10 enterococcal plasmids studied here that encode resistance to chloramphenicol.
PMCID: PMC192749  PMID: 8109922
25.  Effect of clindamycin on intracellular replication, protein synthesis, and infectivity of Toxoplasma gondii. 
Antimicrobial Agents and Chemotherapy  1993;37(12):2571-2577.
We studied the effects of clindamycin and a combination of clindamycin and pyrimethamine on the proliferation of Toxoplasma gondii in cultured mammalian cells and the effect of clindamycin on the parasite's RNA and protein syntheses. Infected macrophages were treated for 48 h with clindamycin or a combination of clindamycin and pyrimethamine, and the 50% inhibitory concentrations for parasite growth were 32.50 +/- 1.30 and 10.78 +/- 0.56 micrograms/ml, respectively. A modified susceptibility assay was also used to measure the effect of low concentrations of clindamycin on T. gondii. Macrophages and bovine turbinate cells were infected with low numbers of tachyzoites and were exposed to low concentrations of clindamycin for 5 days. In these systems, a concentration of 10 ng of clindamycin per ml inhibited 50% of the growth of the parasite in macrophages, while it completely prohibited the growth of the parasite in epithelial cells. When free tachyzoites were preexposed to clindamycin for 4 h, the reduction of parasite infectivity was proportional to the amount of drug; 100 ng of clindamycin per ml reduced the infectivity of T. gondii to 46.5% +/- 8.5% of that of the untreated control. A concentration of 40 micrograms of clindamycin per ml reduced protein synthesis by 56.2% +/- 6.0% but had no effect on RNA synthesis after a 4-h exposure of free tachyzoites of T. gondii to the drug. Our results show that long-term exposure to low concentrations of clindamycin reduces the level of replication of T. gondii, that clindamycin affects the protein synthesis of free parasites, and that clindamycin impairs the ability of tachyzoites to infect host cells.
PMCID: PMC192741  PMID: 7509143

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