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PubMed1999

Drug-resistant Streptococcus pneumoniae in the Lebanon: implications for presumptive therapy.

A total of 50 consecutive clinical isolates of Streptococcus pneumoniae, collected between 1996 and 1998, were tested against six antimicrobial agents using the E-test. The percentages of fully resistant (R) and intermediately-R strains, respectively, were: benzyl penicillin 18 and 38%, amoxycillin-clavulanate 6 and 12%, cefuroxime 22 and 16%, ceftriaxone 2 and 16%, and clarithromycin 10%. Fully and or intermediately multidrug-resistance (two or more drugs) was seen in 44% of the isolates, 18% being fully resistant. The MIC breakpoint for cefaclor is not defined by the National Committee for Clinical Laboratory Standards (NCCLS) but MICs showed that: 76% of the isolates had an MIC of < or = 8 mg/l, 4% had an MIC of 16 mg/l and 20% had an MIC of > or = 32 mg/l. There was agreement between the E-test Pen MIC results and the 1 microg oxacillin (oxa) disk diffusion screen test for the 22 susceptible and the nine fully R strains but not for the 19 strains with Pen MICs between 0.1 and 1 mg/l; this shows the importance of MIC determination in such isolates. Penicillin and multiply antibiotic-resistant pneumococci are spreading in Lebanon, emphasizing the necessity to reconsider current treatment regimens in this country.

Anti-Bacterial Agents↗
PubMed1997

Penicillin-binding protein sensitive to cephalexin in sporulation of Bacillus cereus.

Cephalexin, cefaclor, cefadroxil, and cefotaxime strongly inhibited sporulation of Bacillus cereus ts-4 at 1 microgram/ml. Cephalexin was most inhibitory on the sporulation of B. cereus when the antibiotic was added at 3 h after induction of sporulation by nutrient downshift technique. Examination of 4',6-diamidino-2-phenylindole-stained cells by fluorescence-phase contrast microscopy showed that cephalexin inhibited the formation of asymmetric septum. By using [3H]penicillin, eight penicillin-binding proteins (PBPs) were detected from the cells of B. cereus ts-4. Among them, four PBPs were also detected in sporulating cells. Affinity of cephalexin to PBPs were measured indirectly by competition for subsequent binding of radioactive penicillin G. Cephalexin strongly bound to PBP 4 with molecular weight of 72,000 in sporulating cells.

Bacillus cereus↗
PubMed1996

Management of otitis media with effusion.

Timmy, a two-year-old, has received two antibiotic courses (ampicillin, cefaclor) for acute otitis media during the past year. Today, his mom visits your pharmacy and mentions that they have just returned from an appointment with the boy's doctor. Timmy was diagnosed with otitis media with effusion, which the doctor described as fluid in the ears unaccompanied by fever or any signs of pain or discomfort. She is puzzled that the doctor did not prescribe any antibiotics this time. Worried about her son's recurrent ear problems, she asks your advice. Your review of Timmy's history reveals that he is otherwise healthy except for the two previous episodes of acute otitis media. Further discussion with Timmy's mom also reveals that his father is a heavy smoker.

Child, Preschool↗
PubMed2003

How can we predict bacterial eradication?

Antimicrobial efficacy is measured in vitro by determination of minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of antimicrobials, but these values do not account for fluctuations of drug concentrations within the body or the time course of the drug's in vivo antibacterial activity. However, in vivo bacteriologic efficacy can be predicted by pharmacokinetic/pharmacodynamic (PK/PD) parameters, such as the time for which the serum drug concentration is above the MIC (T>MIC), the ratio of peak serum concentration to the MIC, and the ratio of the area under the 24-h serum concentration-time curve to the MIC (AUC/MIC). Different patterns of antibacterial activity correlate with different PK/PD parameters. For example, a T>MIC of 40-50% of the dosing interval is a good predictor of bacteriologic efficacy for penicillins, cephalosporins, and most macrolides, and an AUC/MIC ratio of at least 25 is required for efficacy with fluoroquinolones and azalides. The PK/PD breakpoint for susceptibility of an organism to a specific dosing regimen of an agent can be determined as the highest MIC met by the relevant PK/PD parameter for bacteriologic efficacy for that agent. These parameters have been validated extensively in animal models, as well as in many human studies where bacteriologic outcome has been determined. The PK/PD breakpoint of an agent is determined primarily by the dosing regimen, and generally applies to all pathogens causing disease at sites where extracellular tissue levels are similar to non-protein-bound serum levels. On this basis, many parenteral beta-lactams are active against almost all strains of Streptococcus pneumoniae, including 'penicillin-non-susceptible' strains, in all body sites except for the central nervous system. Application of PK/PD breakpoints to standard dosing regimens of oral beta-lactams predicts that agents such as cefaclor and cefixime will have efficacy only against penicillin-susceptible strains of S. pneumoniae, while cefuroxime axetil, cefpodoxime and cefdinir will be effective against all penicillin-susceptible as well as many penicillin-intermediate strains. However, the most active oral beta-lactams, amoxicillin and amoxicillin-clavulanate, have predicted efficacy against all penicillin-susceptible and -intermediate pneumococci, as well as against most penicillin-resistant strains, at amoxicillin doses of 45-90 mg/kg per day in children and 1.75-4.0 g/day in adults. These predictions are supported by evidence from animal studies of bacteriologic efficacy. The use of PK/PD parameters to predict bacterial eradication therefore allows an evidence-based approach to the selection of appropriate antimicrobial therapy.

Animals↗
PubMed1985

Bacteriological features and chemotherapy of adult acute purulent otitis media.

Eighty-eight patients from 16 to 79 years old, with acute purulent otitis media, were bacteriologically examined at the Otorhinolaryngology Department of a primary care hospital in Tokyo from July 1979 to May 1983. Fifty-six patients underwent paracentesis, and 32 patients exhibited otorrhea due to previous spontaneous perforation of the tympanic membrane. Bacteriologic cultures revealed the presence of Streptococcus pneumoniae (62.5 per cent), including S. pneumoniae Type III (28.1 per cent), Haemophilus influenzae (10.5 per cent), Staphylococcus aureus (11.5 per cent), and Streptococcus pyogenes (7.3 per cent). S. pneumoniae Type III had a notably high detection rate in patients from 50 to 79 years old (50-75 per cent). Because Haemophilus influenzae was detected at a relatively high rate in patients of all ages, if can be considered as a major causative pathogen of AOM. In 44 patients, selected mainly from those who underwent paracentesis, a comparative study of bacteria found in middle ear fluid and naso-pharyngeal mucus revealed the same bacteria in 43 out of 44 cases (97.7 per cent), indicating the presence of bacterial infection through the auditory canal. Antibiotics were selected according to an Expected Efficacy Index (EEI), the antibiotic of first choice being Ampicillin or Cefaclor.

Acute Disease↗
PubMed1995

Antimicrobial management of chronic sinusitis in children.

This study retrospectively investigated the microbiology and management of 40 children who suffered from chronic sinusitis. The sinuses infected were the maxillary (15 cases), ethmoid (13), and frontal (seven). Pansinusitis was present in five patients. All aspirates were cultured for aerobic and anaerobic bacteria. A total of 121 isolates (97 anaerobic and 24 aerobic) were recovered. Anaerobes were recovered from all 37 culture-positive specimens, and in 14 cases (38 per cent) they were mixed with aerobes. Twenty-three beta-lactamase-producing bacteria were isolated from 16 (43 per cent) patients. The 15 patients who received clindamycin had the most rapid response to therapy and a change of therapy and surgical drainage was required in one case. Of the 16 patients who received amoxycillin or ampicillin, 16 responded to therapy, six needed a change of therapy, including four who also had surgical drainage. Of the six who were treated with erythromycin, three needed antibiotic change, two with surgical drainage. Of the three that received cefaclor, two were cured, and one had an antibiotic change. Resistant organisms were recovered in all the cases that required therapeutic change. These findings support the important role of anaerobic bacteria in the polymicrobial cause of chronic sinusitis in children, and the superiority of therapy effective against these organisms.

Adolescent↗
PubMed2000

The prevalence and clonal diversity of penicillin-resistant Streptococcus pneumoniae in Kuwait.

Penicillin-resistant Streptococcus pneumoniae (PRSP) is widespread all over the world, including countries previously free of PRSP. This study was undertaken to determine the prevalence, the common serotypes and the clonality of PRSP isolated over a period of 1 year, from various clinical samples from three major hospitals in Kuwait. Strains were identified by standard methods and their antibiotic susceptibility was determined by the agar dilution method. The clonality of the isolates was determined by repetitive extragenic palindromic sequence polymerase chain reaction (REP-PCR) genomic profiling and pulsed field gel electrophoresis (PFGE). Serotyping was done by Quellung reaction using specific antisera. We found that 55% of the S. pnuemoniae were resistant to penicillin (46% and 9% exhibited intermediate and full resistance, respectively). Nearly 41% were resistant to sulfamethoxazole-trimethoprim, 9% to cefotaxime and ceftriaxone, 15% to amoxycillin-clavulanate, 17% to cefuroxime, 77 % to cefaclor, and 14% to clindamycin. The commonest serotypes among the PRSPs were 6A, 6B, 14, 19F, 23F and nontypable. PFGE and REP-PCR patterns showed a large diversity of genetic clones of the PRSP. Serotypes 6B, 14, 19F and 23F were more clonally related than the others. Our data showed that the prevalence of PRSP was high, the serotypes were diversified and different genetic clones make up the population of circulating PRSP in Kuwait.

DNA, Bacterial↗
PubMed1992

Antibiotic sensitivities of Neisseria meningitidis isolates from patients and carriers in Greece.

Usage of antibiotics in southern Europe is less well regulated than in northern countries. The proportion (48%) of meningococci in Spain insensitive to pencillin (MIC greater than or equal to 0.1 mg/l) prompted this investigation of antibiotic sensitivities of isolates from Greek patients with meningitis (31) and carriers (47 school-children and 472 recruits). The agar dilution method was used to determine MIC to penicillin G (PN), sulphamethoxazole (SU), rifampicin (RF), cefaclor (CF) and ciprofloxacin (CP). The proportion of isolates insensitive to PN was 48% for isolates from patients, 19% from school-children and 36.6% from recruits. Resistance to SU (MIC greater than or equal to 16 mg/l) was found in 16% of those from patients, 10.6% from children and 40% from recruits. None of the isolates from patients was resistant to RF (greater than or equal to 1 mg/l) but 6% of those from carriers were. Resistance to CF (greater than or equal to 4 mg/l) was found in 9.2% of patient isolates, 6.4% from children and 23.7% from recruits. All isolates except one were sensitive to CP (MIC range less than 0.0015-0.125 mg/l). Resistances to PN, SU and RF were analysed by serogroup, serotype and subtype of the bacteria. The proportion of resistant isolates showed some variation between different areas of Greece, but it was not statistically significant.

Adolescent↗
PubMed1988

Nasopharyngeal carriage and antibiotic resistance of Haemophilus influenzae in healthy children.

An investigation was undertaken to determine the isolation rate and antibiotic resistance of Haemophilus influenzae from the nasopharynx of young children. The 996 subjects studied were up to 6 years of age. H. influenzae was isolated from 304 (30.5%) and strains of capsular type b from 11 (1.1%). Age, sibling status, season, respiratory infection and antibiotic therapy all influenced isolation rates. The overall prevalence of antibiotic resistance in the strains isolated was ampicillin 5.4% (all beta-lactamase producers), cefaclor 0.3%, chloramphenicol 1.3%, erythromycin 38.2%, tetracycline 1.3%, trimethoprim 5.4% and sulphamethoxazole 0%. Ampicillin resistance was more common in type b than non-capsulated strains.

Aging↗
PubMed2006

Binding behavior of subphthalocyanine-tagged testosterone with human serum albumin at the n-hexane/water interface.

Subphthalocyaninatoboron(III) was applied for the first time as a novel marker-tag of testosterone (Subpc-test) for the binding analysis with human serum albumin (HSA) at a liquid/liquid interface. The binding interaction of Subpc-test with HSA at the n-hexane/water interface was studied by UV-visible absorption and circular dichroism (CD) spectroscopy combined with a centrifugal liquid membrane cell at different pHs of aqueous solutions. Complementary studies by a high-speed stirring experiment and an interfacial tension measurement were also performed to characterize the interfacial adsorptivity of Subpc-test and HSA molecules, respectively. The n-hexane solution of Subpc-test showed no optical chirality, but the contact with the aqueous solution of HSA induced its optical chirality, clearly suggesting the formation of Subpc-test/HSA complexes at the n-hexane/water interface. Furthermore, pH profiles of CD signals showed that the interaction between Subpc-test and HSA was very sensitive to the neutral-to-base transition (N-B transition). Interfacial formation of the Subpc-test/HSA complex was studied in the presence of the site-selective ligand, furosemide (site I) or cefaclor (site II). The experimental results suggested that Subpc-test is bound to site I of the HSA molecule at the n-hexane/water interface. This technique using subphthalocyanine as a tag molecule and the liquid/liquid interface as a two-dimensional nanoreaction field will be useful for the evaluation of the interaction between drugs or hormones and proteins.

Humans↗
PubMed1988

Oral absorption of cephalosporin antibiotics. 1. Synthesis, biological properties, and oral bioavailability of 7-(arylacetamido)-3-chloro cephalosporins in animals.

A number of 7-(arylacetamido)-3-substituted cephalosporins were prepared and tested in animals for oral absorbability. Bioavailability in mice, rats, dogs, and monkeys was determined after oral or parenteral administration. Oral bioavailability of five compounds selected for more intensive study was generally higher than that of penicillin V in all species tested. The results of ED50 testing against experimental infections in mice generally supported the bioavailability studies. Antibiotic activities were evaluated against Gram-positive and Gram-negative organisms with some derivatives expressing in vitro activity similar to cefaclor. The plasma half-life in rats was relatively short and the plasma curves were strongly influenced by probenecid, indicating rapid renal secretion. Some 7-(arylacetamido)-3-chloro cephalosporins are orally absorbed in animals to a greater extent than penicillin V, and antibacterial agent of proven clinical utility.

Administration, Oral↗
PubMed2005

A theoretical study on the substrate deacylation mechanism of class C beta-lactamase.

The whole reaction of the deacylation of class C beta-lactamase was investigated by performing quantum chemical calculations under physiological conditions. In this study, the X-ray crystallographic structure of the inhibitor moxalactam-bound class C beta-lactamase (Patera et al. J. Am. Chem. Soc. 2000, 122, 10504-10512.) was utilized and moxalactam was changed into the substrate cefaclor. A model for quantum chemical calculations was constructed using an energy-minimized structure of the substrate-bound enzyme obtained by molecular mechanics calculation, in which the enzyme was soaked in thousands of TIP3P water molecules. It was found that the deacylation reaction consisted of two elementary processes. The first process was formation of a tetrahedral intermediate, which was initiated by the activation of catalytic water by Tyr150, and the second process was detachment of the hydroxylated substrate from the enzyme, which associated with proton transfer from the side chain of Lys67 to Ser64O(gamma). The first process is a rate-determining process, and the activation energy was estimated to be 30.47 kcal/mol from density functional theory calculations considering electron correlation (B3LYP/6-31G**). The side chain of Tyr150 was initially in a deprotonated state and was stably present in the active site of the acyl-enzyme complex, being held by Lys67 and Lys315 cooperatively.

Acylation↗
PubMed1995

Transport characteristics of S-1090, a new oral cephem, in rat intestinal brush-border membrane vesicles.

PURPOSE: Elucidating the transport characteristics of S-1090, a new orally active cephalosporin in rat small intestinal brush-border membranes. METHODS: A rapid filtration technique. RESULTS: The uptake of S-1090 was stimulated by an inwardly directed H(+)-gradient, but did not show overshooting uptake. To investigate the transport system, the inhibitory and countertransport effects of various compounds on S-1090 uptake were examined. Although the dipeptides and tripeptides composed of amino acids with aliphatic side chains did not inhibit the uptake of S-1090, those having histidine, proline or tryptophan as the N-terminal amino acid showed an inhibitory effect. Among the oral cephems tested, ceftibuten showed marked inhibition, while cefaclor and cephalexin had no inhibitory effect. Countertransport effects on S-1090 uptake were observed only when the vesicles were preloaded with histidyl peptides such as His-Gly or His-Ala, while other compounds which exhibited inhibition had no countertransport effect. CONCLUSIONS: Based on the above results, there seems to be heterogeneity (multiplicity) in the oligopeptide transport system which may depend on the structure of the N-terminal amino acid. S-1090 may be dominantly transported via a system that recognizes peptides having histidine as the N-terminal amino acid.

Amino Acid Sequence↗
PubMed1999

Intestinal transport of beta-lactam antibiotics: analysis of the affinity at the H+/peptide symporter (PEPT1), the uptake into Caco-2 cell monolayers and the transepithelial flux.

PURPOSE: This study on the intestinal transport of beta-lactam antibiotics was undertaken to investigate the correlation between cellular transport parameters and the bioavailability. METHODS: Transport of 23 beta-lactam antibiotics was characterized by measuring their ability to inhibit the uptake of glycylsarcosine into Caco-2 cells, their uptake into the cells and their total flux across the cell monolayers. RESULTS: Ceftibuten and cyclacillin were recognized by PEPT1 with affinity constants comparable to those of natural dipeptides (K(i) = 0.3 and 0.5 mM, respectively). Cefadroxil, cefamandole, cephradine, cefaclor, cefuroxime-axetil, cefixime, cephalotin, cephalexin and ampicillin also interacted with PEPTI (K(i) = 7-14 mM). In contrast, cefapirin, cefodizime, cefuroxime, cefmetazole, ceftazidime, benzyl-penicillin, ceftriaxone, cefpirome, cefotaxime, cefepime, cephaloridine and cefsulodin displayed no affinity to the transport system (K(i) > 20 mM). The uptake into the cells and the transepithelial flux was highest for those beta-lactam antibiotics, which showed the strongest inhibition of [14C]Gly-Sar transport (p < 0.0001). Exceptions were cefuroximaxetil and cephalotin. CONCLUSIONS: The probability of oral bioavailability for beta-lactam antibiotics is mainly determined by their affinity to PEPTI. A threshold K(i) value of 14 mM with respect to Gly-Sar uptake is required.

Administration, Oral↗
PubMed1994

Characteristics of ceftibuten uptake into Caco-2 cells.

The characteristics of ceftibuten uptake into Caco-2 cells grown in a collagen-coated dish were examined. Ceftibuten showed stereoselective and pH-dependent uptake. The pH-dependency of ceftibuten was more marked than that of cefaclor or cephalexin, but all three antibiotics showed maximal uptake at pH 5.5. Ceftibuten uptake was linear for the initial 1 hr and then reached a plateau. The initial uptake (15 min) was markedly reduced by the addition of 2,4-dinitrophenol or FCCP (a protonophore), or by lowering the incubation temperature. The uptake of ceftibuten into the brush-border membrane vesicles prepared from cultured Caco-2 cells showed an overshoot in the presence of an H(+)-gradient. These findings indicated that the uptake of ceftibuten was energy-dependent, especially H(+)-gradient-dependent. Uptake inhibition by various compounds was compared using Caco-2 cells. Amino acids and a tetrapeptide did not inhibit uptake, whereas di- or tri-peptides were effective inhibitors. These observations suggest that ceftibuten is taken up by a carrier-mediated transport system(s) for dipeptides. Various antibiotics differed in their ability to inhibit uptake, with cyclacillin showing maximum inhibition. Differences in the inhibitory effect may be accounted for by the heterogeneity (multiplicity) of the transport systems.

2,4-Dinitrophenol↗
PubMed2003

Prescription medication use during pregnancy and risk of infant leukemia (United States).

OBJECTIVE: We explored whether maternal medication use during pregnancy may be an important etiologic area for investigation in studies of infant leukemia. METHODS: In this case-control study, associations were explored between specific medications as recorded in the medical records of 243 mothers of infants who were diagnosed with leukemia at < 18 months of age and 393 mothers ol infants without leukemia identified through random digit dialing. Cases included 157 acute lymphoblastic (ALL), 77 acute myeloid (AML), and nine other leukemias. A total of 27 different drugs that were prescribed for at least six women were analyzed. RESULTS: Overall, non-statistically significant negative associations (Odds ratios, OR < 0.5) were observed with amoxicillin, nystatin, clomiphene, levothyroxine, cefaclor, and trimethobenzamide HCL. When cases were restricted to either myeloid or lymphoblastic leukemia, cotrimoxazole was prescribed for five ALL case mothers and no matched controls; no association with cotrimoxazole was observed with AML. CONCLUSIONS: Given the number of comparisons, chance cannot be ruled out for any of the associations found here. However, a strength of this study is the lack of recall bias. The disparate data observed between AML and ALL in some instances may indicate areas of interest; they will be explored further in a case-control study of infant leukemia that is currently underway.

Case-Control Studies↗
PubMed1995

Multiple substitutions at position 104 of beta-lactamase TEM-1: assessing the role of this residue in substrate specificity.

Residue 104 is frequently mutated from a glutamic acid to a lysine in the extended-spectrum TEM beta-lactamases responsible for the resistance to third-generation cephalosporins in clinical Gram negative strains. Among class A beta-lactamases, it is the most variable residue within a highly conserved loop which delineates one side of the active site of the enzymes. To investigate the role of this residue in the extended-spectrum phenotype, it has been replaced by serine, threonine, lysine, arginine, tyrosine and proline. All these substitutions yield active enzymes, with no drastic changes in kinetic properties compared with the wild-type enzyme, except with cefaclor, but an overall improved affinity for second- and third-generation cephalosporins. Only mutant E104K exhibits a significant ability to hydrolyse cefotaxime. Molecular modelling shows that the substitutions have generally no impact on the conformation of the 101-111 loop as the side chains of residues at position 104 are all turned towards the solvent. Unexpectedly, the E104P mutant turns out to be the most efficient enzyme. All our results argue in favour of an indirect role for this residue 104 in the substrate specificity of the class A beta-lactamases. This residue contributes to the precise positioning of residues 130-132 which are involved in substrate binding and catalysis. Changing residue 104 could also modify slightly the local electrostatic potential in this part of the active site. The limited kinetic impact of the mutations at this position have to be analysed in the context of the microbiological problem of resistance to third-generation cephalosporins. Although mutation E104K improves the ability of the enzyme to hydrolyse these compounds, it is not sufficient to confer true resistance, and is always found in clinical isolates associated with at least one mutation at another part of the active site. It is the combined effect of the two mutations that synergistically enhances the hydrolytic capability of the enzyme towards third-generation cephalosporins.

Amino Acid Sequence↗
PubMed2000

Modification of general practitioner prescribing of antibiotics by use of a therapeutics adviser (academic detailer).

AIMS: This was a pilot study of the use of a clinical pharmacist as a therapeutics adviser (academic detailer) to modify antibiotic prescribing by general practitioners. METHODS: Following a visit by the adviser (March-May), 112 general practitioners were recruited and randomised to control or active groups. A panel of experts prepared a best practice chart of recommended drugs for upper and lower respiratory tract infections, otitis media and urinary tract infections. The adviser made a 10-15 min visit to each prescriber in the active group (June-July), gave them the chart and discussed its recommendations briefly. Doctors in the control group were not visited nor given the chart. Prescription numbers for all prescribers were obtained from the Commonwealth Health Insurance Commission for the pre(March-May) and postdetailing (August-September) periods using a three month lag time for data collection. Data for total numbers of prescriptions and for selected individual antibiotics used in these two periods were analysed using nonparametric statistics. RESULTS: Prescribing patterns were similar for the control and active groups in the predetailing period. For both groups, there were significant (P<0.03) increases (45% for control and 40% for active) in total number of antibiotic prescriptions in the post compared with the predetailing period. This trend was anticipated on the basis of the winter seasonal increase in respiratory infections. In line with the chart recommendations for first-line treatment, doctors in the active group prescribed significantly more amoxycillin (P<0.02) and doxycycline (P<0.001) in the post vs predetailing periods. By contrast, doctors in the control group prescribed significantly more cefaclor (P<0.03) and roxithromycin (P<0.03), drugs that were not recommended. The total cost of antibiotics prescribed by doctors in the control group increased by 48% ($37 150) from the preto postdetailing periods. In the same time period, the costs for the active group increased by only 35% ($21 020). CONCLUSIONS: We conclude that the academic detailing process was successful in modifying prescribing patterns and that it also decreased prescription numbers and costs. Application of the scheme on a nationwide basis could not only improve prescriber choice of the most appropriate antibiotic but also result in a significant saving of health care dollars.

Anti-Bacterial Agents↗