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PubMed1999

Cell wall-affecting antibiotics induce expression of a novel gene, drp35, in Staphylococcus aureus.

A novel gene, drp35, of Staphylococcus aureus, which was inducible especially with cell wall-affecting antibiotics, has been cloned. Analysis of differential hybridization with mRNAs enhanced in the presence of beta-lactams resulted in two positive clones that harbored a new gene encoding a 35,845-Da protein (Drp35) and the penicillin-binding protein 2 (PBP2). Immunoblot analysis revealed that the Drp35 protein band was evidently enhanced after 30 min in the presence of beta-lactams. The Drp35 expression was also enhanced with not only beta-lactams, but also vancomycin, bacitracin, and fosfomycin. Homology search revealed that Drp35 was a new protein. Our results revealed that it was specific in S. aureus and respondent to these agents in both methicillin-resistant and -sensitive strains of S. aureus.

Amino Acid Sequence↗
PubMed1988

Nontoxic concentration of antibiotics for intravitreal use--evaluated by human in-vitro ERG.

The effects of penicillin G (PC-G) sodium, procaine PC-G, cloxacillin sodium (MCIPC), disodium sulbenicillin (SBPC), cefazolin sodium (CEZ), gentamicin sulfate (GM) and fosfomycin sodium (FOM) on the electroretinogram (ERG) of the human in-vitro eye-cup were studied. The oscillatory potentials (OPs) were selectively and greatly suppressed by 1.0 mM PC-G sodium. The OPs and c-wave were suppressed by 0.85 mM procaine PC-G. The b-wave and OPs were slightly suppressed by 1.0 mM MCIPC. The a-wave, b-wave, OPs and c-wave were not deteriorated by 1.0 mM SBPC. The OPs appeared to be selectively suppressed by 1.0 mM CEZ. The b-wave was suppressed and the peak latencies of the OPs were delayed by 184 micrograms/ml (approximately 0.4 mM) GM. The amplitudes of the a-wave and c-wave were slightly enhanced and their peak latencies were slightly delayed by 184 micrograms/ml GM. The a-wave, b-wave, OPs and c-wave were not deteriorated by 1.0 mM FOM. The results of the present study on the human retina were comparable to those on the albino rabbit retina in our previous studies.

Anti-Bacterial Agents↗
PubMed1994

A simple identification scheme for coagulase negative staphylococci from bovine mastitis.

Coagulase-negative staphylococci from cases of bovine mastitis were identified to species level by using an identification scheme based on a three-plate test system which tested for DNase on DNA agar, for protease on calcium caseinate agar, and for the organism's sensitivity to novobiocin, desferrioxaminine (deferoxamine) and fosfomycin by agar diffusion tests. Testing for the inhibition of Staphylococcus delta haemolysin (Skalka 1991) can replace the protease tests.

Agar↗
PubMed1983

The micronucleus test with mouse spleen cells.

The results of this study show that the micronucleus test can be carried out with mouse spleen cells as well as with cells from bone marrow. Polychromatic erythrocytes occurred in the spleen at a frequency of about 9% of the whole spleen cells compared with about 13% in the bone marrow. 3 test compounds were used to compare the frequency of micronuclei in cells from the 2 tissues. Mitomycin C and cyclophosphamide induced micronucleated polychromatic erythrocytes in both spleen and bone marrow. Fosfomycin, an antibiotic having a broad spectrum of antimicrobial activities, did not induce micronucleated erythrocytes in either organ.

Animals↗
PubMed2005

Suspected disk space infection with negative microbiological tests-report of eight cases and comparison with documented pyogenic discitis.

UNLABELLED: Few data are available on patients with suspected noniatrogenic pyogenic discitis but negative microbiological tests. OBJECTIVES: To compare the features, treatment, and outcomes in patients with suspected versus microbiologically documented noniatrogenic pyogenic discitis. PATIENTS: A retrospective chart review identified eight patients with suspected noniatrogenic pyogenic discitis managed at our institution over a 15-year period. Eighteen age- and sex-matched patients with microbiologically documented noniatrogenic pyogenic discitis managed at our institution during the same period served as controls. RESULTS: The eight cases had a longer time to diagnosis, a greater risk of abscess formation or epidural infection, and a smaller number of cutaneous portals of entry, as compared to the controls. None of the cases had evidence of endocarditis or diabetes mellitus. Infection at another site preceded the discitis in half the cases. Empirical treatment with two antimicrobials (usually a fluoroquinolone with a beta-lactam or fosfomycin) ensured a full recovery in all eight cases, with no relapses or long-term recurrences, whereas relapses occurred in three of the 18 controls treated with antibiotics selected by antibiotic susceptibility testing. CONCLUSION: Noniatrogenic pyogenic discitis with negative microbiological studies is associated with distinctive clinical features consistent with smoldering infection. The outcome is favorable under empirical two-drug antimicrobial therapy, usually including a fluoroquinolone given by the intravenous route.

Abscess↗
PubMed2002

Urinary tract infection: traditional pharmacologic therapies.

Urinary tract infections (UTIs) are common bacterial infections, particularly in women. Antimicrobial therapy is seldom indicated for asymptomatic infection, but antimicrobial therapy is usually indicated for amelioration of symptoms. Management of acute uncomplicated UTI (cystitis) is generally straightforward, with a predictable distribution of uropathogens isolated. First-line treatment of acute uncomplicated UTI has traditionally involved a 3-day regimen of trimethoprim-sulfamethoxazole (TMP-SMX) or TMP alone for patients with sulfa allergies. Increasing resistance among community-acquired Escherichia coli to TMP-SMX worldwide has led to a reassessment of the most appropriate empiric therapy for these infections. Alternative first-line agents include the fluoroquinolones, nitrofurantoin, and fosfomycin. Factors to be considered in the selection of appropriate antimicrobial therapy include pharmacokinetics, spectrum of activity of the antimicrobial agent, resistance prevalence for the community, potential for adverse effects, and duration of therapy. Ideal antimicrobial agents for UTI management have primary excretion routes through the urinary tract to achieve high urinary drug levels. In addition, there are special considerations in the management of UTI among selected populations, including postmenopausal and pregnant women, and for women with frequent recurrent UTIs.

Adolescent↗
PubMed2004

[Suppurative cervical lymphadenitis in children. Review of 45 patients].

OBJECTIVE: To review the bacteriological features and treatment of suppurative cervical lymphadenitis in children in order to achieve optimal treatment. PATIENTS AND METHODS: Forty-five children were hospitalized. Time of hospitalization was evenly distributed over Year and between Years. Age ranged from 2.5 Months to 13.5 Years (median 1 Year 10 Months). Twenty-nine patients had received prior antibiotic treatment. Bacteriological samples were sterile in 15 out of 45 children and were positive for Staphylococcus (all meti-S) in 20. Cefotoxim and fosfomycin were prescribed and was sufficient in 14 children (with needle aspiration). Surgical drainage was performed in the other cases. CONCLUSION: Staphylococcus aureus is the predominant causal agent for acute suppurative lymphadenitis in children. Antibiotic therapy in the outpatient setting does not always prevent abscess formation and surgical drainage may be required.

Acute Disease↗
PubMed2002

[Rhône-Alpes observatory of Streptococcus pneumoniae in 1999: 35 cases of meningitis].

In 1999, in Rhône-Alpes region, in a survey of resistance to antibiotics of Streptococcus pneumoniae, 35 cases of meningitis were observed. A retrospectic questionnary was sent to each participant. MICs to Penicillin, Amoxicillin and Cefotaxime were determined with ATB-PNEUMO gallery or E-test and by disk diffusion for the other antibiotics. The results were interpreted according to the recommendations of the CA-SFM. Mean age was 38.1 years (range : 1 month -78 years) and sex-ratio 2/5. Eight patients had previously received antibiotics, 22 patients had risk factors and 23 were transferred in intensive care unit. The patients received C3G + glycopeptide in 15 of 16 children and in 13/19 adults according to the consensus recommendations. Diagnostic was made on the direct examination of CSF in 83%, and blood cultures was positive in 74.3% of cases. The percentage of PRP was 48.6% with 17.1% of intermediate-amoxicilline and 14.3% intermediate-cefotaxime strains. Resistance to trimethoprim-sulfamethoxazole was 45.7%, to chloramphenicol 30% and to fosfomycin 6.9%. All the strains were susceptible to rifampicin and vancomycin. Among the 17 PRP strains, 7 were belonging to serotype 6 (6 in children). The clinical outcome was fatal in 7 male cases (20%), without risk factors in 3 children and 6 of 7 strains were susceptible to penicillin. Six patients (17%) had auditive and/or neurologic sequellaes. This study shows that nearly 50% of strains isolated in meningitis, in Rhône-Alpes region, were not susceptible to penicillin, and confirms the frequency of sequellaes while the mortality is not related with the resistance of strains to the antibiotics.

Adolescent↗
PubMed2000

Macrolides and clindamycin suppress the release of Shiga-like toxins from Escherichia coli O157:H7 in vitro.

We investigated the effects of antimicrobial agents fosfomycin (FOS), cefdinir (CDIN), levofloxacin (LEVX), rokitamycin (ROK), roxithromycin (ROX), and clindamycin (CLI) on the release of Shiga-like toxins (SLTs) by enterohaemorrhagic Escherichia coli (EHEC). EHEC was cultured for 14 h in the presence of ROX, ROK or CLI at sub-minimum inhibitory concentrations (subMICs) of 1.56-6.25 mg/l, followed by assay of the level of SLTs in the supernatants using cytotoxicity assay and reversed passive latex agglutination method. Exposure to ROX, ROK or CLI reduced the amount of released SLTs compared with untreated control cultures (P<0.05). These agents however, did not decrease the number of viable EHEC, indicating the importance of bactericidal agents. When the bacteria was exposed to CDIN, FOS or LEVX, the level of SLTs in the culture supernatant increased with the destruction of bacterial cells in the order of CDIN, FOS, LEVX. When 0.5 mg/l of LEVX was added to cultures with or without pretreatment using ROX, ROK, or CLI, the release of SLTs was reduced by this pretreatment (P<0.05). These results may have clinical implications for the treatment of EHEC infection.

Animals↗
PubMed2002

Modulatory effect of macrolide antibiotics on the Th1- and Th2-type cytokine production.

The effect of the macrolide antibiotics, clarithromycin, midecamycin acetate and josamycin, on the generation of Th1- and Th2-type cytokines by mitogen-stimulated human T lymphocytes was compared with that of fosfomycin. The following results were obtained. These drugs demonstrated potent inhibitory activity on the release and gene expression of TNF-alpha and IL-2. Their inhibitory effect on IFN-alpha, IL-4, IL-5, IL-6 was less marked. The release of IL-10 was poorly suppressed. Clarithromycin had the most potent inhibitory effect of the drugs used. The present results suggested that anti-bacterial agents might modify the host's immunological response by interfering with the activity of T helper cells.

Anti-Bacterial Agents↗
PubMed1998

[Severe streptococcal group A infection complicating varicella].

UNLABELLED: Varicella is a common viral infection which is generally benign in infancy and has a good outcome. It may sometimes be complicated by severe group A streptococcal superinfection. CASE REPORT: Three days after the beginning of varicella, a previously healthy 2-year-old girl presented with left leg pain, lameness and edema of all four limbs. Toxic shock syndrome occurred, due to beta-hemolytic group A Streptococcus grown from blood culture. Computerized tomography (CT) scan showed a mild effusion involving both hips. Cefotaxim was administered, but the week after magnetic resonance imaging (MRI) showed a necrotizing fasciitis and a lesion of the left leg leading to a patchy femoral diaphysis consistent with osteomyelitis. Joint aspirate culture did not grow. The left leg was immobilized in plaster for 6 weeks and the child was given cefotaxim and fosfomycin parenterally during 30 days, then followed by 45 days of oral amoxicillin. She recovered without sequelae. CONCLUSION: Group A Streptococcus infection is a dangerous complication of varicella. It must be considered in case of any joint pain occurring during or just after this disease. The choice of the best treatment needs full collaboration between surgeons, radiologists and pediatricians.

Cefotaxime↗
PubMed1998

Mechanistic imperatives for the evolution of glutathione transferases.

Several significant advances in the understanding of the catalytic mechanisms, structures and evolution of glutathione transferases have occurred in the past year. These advances include new mechanistic information concerning the canonical soluble enzymes, the finding that the fosfomycin-specific enzyme, FosA, is a metalloglutathione transferase and a higher resolution projection structure of the microsomal enzyme.

Bacterial Proteins↗
PubMed2001

Asparagine 23 and aspartate 305 are essential residues in the active site of UDP-N-acetylglucosamine enolpyruvyl transferase from Enterobacter cloacae.

UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) catalyzes the transfer of the intact enolpyruvyl moiety of phosphoenolpyruvate (PEP) to the 3'-hydroxyl group of UDP-N-acetylglucosamine (UDPNAG). This reaction constitutes the first committed step in the biosynthesis of the bacterial cell wall component peptidoglycan (murein). The transfer reaction involves the nucleophilic attack of the 3'-hydroxyl group of UDPNAG at the C-2 of PEP. The three-dimensional structure of MurA complexed with UDPNAG revealed an aspartate residue (D305 in the En. cloacae sequence) close to the 3'-hydroxyl group of UDPNAG, suggesting that it may act as an acid-base catalyst in the active center of the enzyme. In addition to aspartate 305, asparagine 23 also interacts with the 3'-hydroxyl group; however, its role in catalysis or binding of the UDPNAG substrate is unclear. To gain information on the role of these two amino acids in the MurA-catalyzed reaction we have exchanged D305 for alanine, cysteine, histidine, and glutamate, and N23 for alanine and serine using site-directed mutagenesis. While the D305 alanine, cysteine, and histidine mutant proteins do not have detectable enzymatic activity, the D305E mutant protein exhibits a low residual activity (ca. 0.1% of the wild-type enzyme). Unlike with wild-type MurA, no exothermic signal was obtained when the D305A and -E mutant proteins were titrated with UDPNAG, demonstrating that the affinity of the sugar nucleotide substrate is reduced as a result of the amino acid exchange. The reduced affinity to UDPNAG leads to a lower propensity of C115 to form either the O-phosphothioketal with PEP or the thioether with the antibiotic fosfomycin. These findings emphasize the dual role of D305 as a general base and an essential binding partner to UDPNAG in the active site of MurA. Similarly, the two N23 mutant proteins showed a much lower catalytic activity although binding of UDPNAG was not as much affected as in the case of the D305 mutant proteins. This result indicates that this amino acid residue is mainly involved in stabilization of transition states.

Alkyl and Aryl Transferases↗
PubMed2000

Mechanistic diversity in a metalloenzyme superfamily.

It is now appreciated that the relationships of proteins, particularly enzymes, within a protein superfamily can be understood not only in terms of their sequence similarities and three-dimensional structures but also by chemical threads that relate their functional attributes. The mechanistic ties among superfamily members can often be traced to a common transition state for the rate-limiting step of the reactions being catalyzed. This paper presents an analysis of a metalloenzyme superfamily, the members of which catalyze a very diverse set of reactions with unrelated transition states but a more general common mechanistic imperative. The vicinal oxygen chelate (VOC) superfamily is composed of structurally related proteins with paired beta alpha beta beta beta motifs that provide a metal coordination environment with two or three open or readily accessible coordination sites to promote direct electrophilic participation of the metal ion in catalysis. The known types of reactions that are catalyzed include isomerizations (glyoxalase I), epimerizations (methylmalonyl-CoA epimerase), oxidative cleavage of C-C bonds (extradiol dioxygenase), and nucleophilic substitutions (fosfomycin resistance proteins). The remarkable access to mechanism space that is provided by the VOC superfamily appears to derive from a simple, pseudosymmetric structural fold that maximizes the catalytic versatility of the metal center.

Acetaldehyde↗
PubMed2004

Control of bacteria adhesion by cell-wall engineering.

UDP-MurNAc-pentapeptide derivative bacterial cell-wall precursors were synthesized as effective tools for surface display on living bacteria. Lactobacilli were incubated in the ketone-modified precursor-containing medium, and the ketone moiety was displayed on the bacterial surface through cell-wall biosynthesis. Oligomannose was coupled with the ketone moiety on the bacterial surface via a aminooxyl linker, thereby displaying this oligosaccharide on the surface of the bacteria. The increase in the adhesion of the sugar-displaying bacteria onto a concanavalin A-attached film compared to that of native bacteria was confirmed by microscopic observation and surface plasmon resonance measurement. The incorporation of the artificial cell-wall precursors was enhanced when incubated with fosfomycin, an inhibitor of cell-wall precursor biosynthesis.

Anti-Bacterial Agents↗
PubMed2006

Rings, radicals, and regeneration: the early years of a bioorganic laboratory.

This Perspective provides an overview of the progress in two of the original programs in my research group focused on the biosynthesis of the antibiotics nisin, lacticin 481, fosfomycin, and bialaphos. The path from start-up funds to tenure and beyond offers insights into the opportunities realized and missed along the road.

Bacteriocins↗
PubMed2003

Two-component system VraSR positively modulates the regulation of cell-wall biosynthesis pathway in Staphylococcus aureus.

DNA microarray covering the whole genome of Staphylococcus aureus strain N315 was prepared to investigate transcription profiles. The microarray analyses revealed that vancomycin induces transcription of 139 genes. Forty-six genes among them failed to be induced in the vraSR null mutant KVR. Part of the genes regulated by VraSR system is associated with cell-wall biosynthesis, such as PBP2, SgtB and MurZ. Other cell-wall synthesis inhibitors also induced VraSR, suggesting that the sensor kinase VraS responds to the damage of cell-wall structure or inhibition of cell-wall biosynthesis. Additionally, the vraSR null mutants derived from hetero- and homo-methicillin-resistant S. aureus showed significant decrease of resistance against teicoplanin, beta-lactam, bacitracin and fosfomycin but not of D-cycloserine and levofloxacin. The observation strongly indicates that VraSR constitutes a positive regulator of cell-wall peptidoglycan synthesis, and that is deeply involved in the expression of beta-lactam and glycopeptide resistance in S. aureus.

Anti-Bacterial Agents↗
PubMed2000

Enterocolitis with pathogenic Escherichia coli infection in renal transplant recipients: case reports.

PURPOSE: We report three cases of enterocolitis associated with pathogenic Escherichia coli infection in renal transplant recipients. METHODS/RESULTS: Patients presented with abdominal pain and diarrhea at 1, 3 and 7 years after living-related renal transplantation. Pathogens of enterocolitis were identified by stool culture as verotoxin-2-producing O157:H7 E. coli, non-verotoxin-producing E. coli 06 and 0125. All patients were basically treated with fluid replacement with additional fosfomycin administration in the patient with O157:H7 E. coli infection. Immunosuppressive drugs were kept at maintenance doses throughout the treatment. CONCLUSION: All patients recovered uneventfully within 10 days after the onset of enterocolitis without severe complications.

Adult↗