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In vitro and in vivo studies of three antibiotic combinations against gram-negative bacteria and Staphylococcus aureus.

The activities of azlocillin, cefotaxime, and amikacin alone and in combination were evaluated in in vitro checkerboard studies, in infected neutropenic mice, and in human volunteers. The combination of cefotaxime plus amikacin was more synergistic in vitro than the others against the Enterobacteriaceae tested, and the combination of azlocillin plus amikacin was more synergistic against Pseudomonas aeruginosa and Staphylococcus aureus. Survival of neutropenic mice infected with Escherichia coli and Klebsiella pneumoniae, respectively, was greater with azlocillin plus amikacin (24 of 40 and 11 of 40) and with cefotaxime plus amikacin (21 of 40 and 17 of 40) than with azlocillin plus cefotaxime (22 of 40 and 3 of 40; P less than 0.05). Median serum bactericidal activity in volunteers receiving these antibiotics alone and in combination was greater than or equal to 1:8 with most agents and with all combinations tested against 10 strains each of E. coli, K. pneumoniae, P. aeruginosa, and S. aureus. These data suggest that clinical trials with combinations of azlocillin or cefotaxime plus amikacin deserve further study in febrile neutropenic patients.

Amikacin↗

Comparative in vitro antibacterial activity of seven semi-synthetic penicillins against aerobic gram-negative bacteria and enterococci.

The MICs and MBCs of mecillinam, ticarcillin, mezlocillin, azlocillin and piperacillin were determined by the microdilution method in liquid medium using 700 strains of gram-negative bacilli and enterococci isolated from pathological sources and classified as a function of their sensitivity to ampicillin and carbenicillin. The ampicillin and carbenicillin-sensitive strains were generally sensitive to the other penicillins, although there were differences in activity. The ampicillin and carbenicillin-resistant strains of Escherichia coli that produce a TEM-type penicillinase were sensitive to mecillinam. Mezlocillin, piperacillin and azlocillin had MICs of between 32 and 64 mg/l for 40% of these strains. The Klebsiella strains, whose broad-spectrum penicillinase deactivates ampicillin and carbenicillin, remained sensitive to mecillinam. Mezlocillin, azlocillin and piperacillin had MICs of less than 8 mg/l for 50% of these strains. The carbenicillin-resistant strains of Enterobacter and Citrobacter were also resistant to the other penicillins. Piperacillin and mezlocillin displayed some activity against certain strains of carbenicillin-resistant Serratia, Proteus and Acinetobacter. Azlocillin, piperacillin and, to a lesser degree, mezlocillin were active against the strains of Pseudomonas, for which carbenicillin had an MIC of about 512 mg/l. Ampicillin, mezlocillin and azlocillin showed the best activity against the enterococci, against which mecillinam was inactive. The MBC of these antibiotics is greatly influenced by the density of the bacterial inoculum.

Gram-Negative Aerobic Bacteria↗

Activity and synergy of ureido penicillins and aminoglycosides against Pseudomonas aeruginosa.

The in vitro activities of piperacillin, azlocillin, mezlocillin, sulbenicillin and ticarcillin were compared with those of carbenicillin using 88 clinical isolates of Pseudomonas aeruginosa. The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) were determined by standard techniques. The MIC for 90% of the strains was 7.5 mg/l for piperacillin, 10.0 mg/l for azlocillin, 26.5 mg/l for mezlocillin, 48.4 mg/l for sulbenicillin, 50.0 mg/l for ticarcillin and more than 100 mg/l for carbenicillin. The MBC/MIC ratio was 1.3 for piperacillin, 1.9 for ticarcillin, 2.1 for sulbenicillin, 3.3 for mezlocillin and 4.5 for azlocillin. The susceptibilities of the same strains to four aminoglycosides were tested. The MIC for 90% of the strains was 0.3 mg/l for sisomicin and tobramycin, 1.5 mg/l for amikacin, and 2.2 mg/l for gentamicin. The effect of combining piperacillin, azlocillin and mezlocillin with gentamicin, tobramycin, sisomicin and amikacin was studied using checkerboard titration. The highest degrees of synergy were found with the combinations of piperacillin and an aminoglycoside. Strong potentiation was observed in 85% of the strains with piperacillin - sisomicin and in 50% with piperacillin - gentamicin. The synergistic effects of azlocillin and mezlocillin in combination with an aminoglycoside (observed in 30-65% of the strains) were for the most part moderate or slight. No antagonism was observed.

Aminoglycosides↗

Comparative study of the binding of acylureidopenicillins and carbenicillin to human serum proteins.

The binding of the acylureidopenicillins azlocillin and mezlocillin to serum proteins was investigated by means of equilibrium dialysis and ultracentrifugation. The penicillin concentrations were determined by the agar diffusion test and by circular dichroism. The degree of binding is dependent on the penicillin concentration. At a mean penicillin concentration of 20 microgram/ml 39% of azlocillin is bound by serum proteins and 35% of mezlocillin. The acylureidopenicillins are bound to about the same degree by the two proteins albumin and gamma globulin, which occur in different concentrations in serum. The binding affinity delta F degree for the interaction between azlocillin and mezlocillin to human albumin is equal to -15,389 J/mol. The corresponding values for the binding of ampicillin and carbenicillin are -17,340 and -16,800 J/mol. Albumin has one binding site of a high affinity and two binding sites of a low affinity for the acylureidopenicillins. Measurements of partition coefficients of the penicillins for isobutanol and aqueous solution show that the interaction between acylureidopenicillins and serum is mainly hydrophobic in nature. This can be deduced from the fact that for example in the presence of sisomicin azlocillin is not displaced from its albumin binding site, in contrast to carbenicillin. The reason for this difference in behaviour lies in the fact that carbenicillin, like sisomicin, is bound to the serum proteins mainly by ionic bonds. In the case of azlocillin, however, hydrophobic bounds predominate which are not influenced by the ionic bonds of sisomicin.

Blood Proteins↗

Clinical pharmacology of extended-spectrum penicillins in infants and children.

Compared with previously available penicillins, piperacillin, azlocillin, and mezlocillin have increased activity in vitro against gram-negative bacilli. After intravenous administration of conventional doses (50 to 100 mg/kg) in children, peak concentrations of these drugs are approximately 70 to 350 micrograms/ml. For piperacillin, azlocillin, and mezlocillin, the half-lives during the beta elimination phase (t 1/2 beta) are approximately 0.5 to 0.75, 0.8 to 1.7, and 0.8 to 1.0 hours, respectively. In patients receiving the higher dosage, particularly of azlocillin, the t 1/2 beta may be prolonged by approximately 20%. A total daily dosage of 300 mg/kg or 9 gm/m2 given in four to six divided dosages should produce peak concentrations of approximately 150 micrograms/ml, and concentrations greater than 16 micrograms/ml for at least 2 hours after each administration. Lower daily dosages are needed in neonates, but precise dosage recommendations cannot be made at this time. Only approximately 60% of piperacillin and approximately 45% of azlocillin are eliminated unchanged in the urine; thus only modest dosage reductions are needed in patients with decreased renal function. In children, adverse effects have been infrequent.

Absorption↗

Comparison of a beta-lactam alone versus beta-lactam and an aminoglycoside for pulmonary exacerbation in cystic fibrosis.

UNLABELLED: We determined whether a beta-lactam and an aminoglycoside have efficacy greater than a beta-lactam alone in the management of a pulmonary exacerbation in patients with cystic fibrosis. STUDY DESIGN: Azlocillin and placebo or azlocillin and tobramycin were administered to 76 patients with a pulmonary exacerbation caused by Pseudomonas aeruginosa in a randomized double-blind, third-party monitored protocol. Improvement was assessed by standardized clinical evaluation, pulmonary function testing, sputum bacterial density, sputum DNA content, and time to the next pulmonary exacerbation requiring hospitalization. RESULTS: No significant difference was seen between the 2 treatment groups in clinical evaluation, sputum DNA concentration, forced vital capacity, forced expiratory volume in second 1, or peak expiratory flow rate at the end of treatment (33 receiving azlocillin alone and 43 both antibiotics); adverse reactions were equivalent in each group. Sputum P. aeruginosa density decreased more with combination therapy (P =.034). On follow-up evaluation, an average of 26 days after the end of treatment, all outcome indicators had worsened in both groups. Time to readmission for a new pulmonary exacerbation was significantly longer in the group receiving azlocillin plus tobramycin (P <.001). Treatment-emergent tobramycin resistance occurred in both groups and was more frequent with combination therapy. CONCLUSION: We conclude that the combination of a beta-lactam and an aminoglycoside produces a longer clinical remission than a beta-lactam alone and slightly better initial improvement.

Adolescent↗

Ciprofloxacin in experimental aortic valve endocarditis due to Pseudomonas aeruginosa.

Left-sided endocarditis caused by Pseudomonas aeruginosa is frequently associated with failure of medical therapy in man. The efficacy of ciprofloxacin and netilmicin + azlocillin has been studied in 79 rabbits with aortic valve endocarditis caused by a serum-resistant strain of P. aeruginosa. Infected animals received either: no therapy; ciprofloxacin (80 mg/kg/day); or netilmicin (6.5 mg/kg/day) + azlocillin (400 mg/kg/day). Ciprofloxacin significantly lowered vegetation titers of P. aeruginosa at days 6 and 10 of therapy compared with netilmicin + azlocillin (P less than 0.001). Similarly, ciprofloxacin was significantly more effective in sterilizing vegetations (P less than 0.005), curing P. aeruginosa endocarditis (P less than 0.001), and preventing bacteriological relapse after discontinuing antibiotic therapy (P less than 0.005). Both antibiotic regimens were equally effective in sterilizing renal abscesses. Resistance to azlocillin was occasionally observed in vivo among P. aeruginosa isolates within cardiac vegetations during the second week of therapy, but not to ciprofloxacin or netilmicin.

Abscess↗

A comparison of double beta-lactam combinations with netilmicin/ureidopenicillin regimens in the empirical therapy of febrile neutropenic patients.

In a randomized trial ceftazidime plus piperacillin or azlocillin, and netilmicin plus piperacillin or azlocillin were used as initial empirical therapy in 202 febrile neutropenic episodes. Netilmicin plus azlocillin was the most effective combination with a clinical response rate of 81% in clinically and microbiologically documented infections compared with 63% for ceftazidime plus piperacillin. All of the episodes of Gram-negative bacteraemia treated with azlocillin responded compared with 43% of those treated with piperacillin. Gram-positive organisms accounted for 52% of all bacteriologically documented infections and 40% of the febrile episodes were treated with vancomycin for presumptive or documented Gram-positive infection. Patients treated with netilmicin had significantly more nephrotoxicity than those given the double beta-lactam combinations (14.8% vs 3.5%; P less than 0.05). However, this difference was not shown in those patients who did not receive concurrent vancomycin or amphotericin. The double beta-lactam combinations were associated with more hypokalaemia (58.2% vs. 37.7%; P less than 0.05) and more colonization with yeasts (24% vs. 10.4%; P less than 0.05) but there was no evidence that their use was associated with prolongation of neutropenia. These results indicate that ceftazidime plus a ureidopenicillin would be adequate empirical therapy in situations where the concomitant use of nephrotoxic agents precludes the use of aminoglycoside containing combinations.

Adult↗

The in vitro effect of three antibiotics on alveolar macrophages.

The effects of 3 antibiotics, azlocillin, ticarcillin and tobramycin, on rat alveolar macrophage function has been examined. These antibiotics are used in the treatment of Pseudomonas aeruginosa lung infections in cystic fibrosis patients. In this study low concentrations of the antibiotics were used, similar to the low levels found in the lungs of these patients. Tobramycin, a highly active aminoglycoside, inhibited the phagocytosis of opsonized Pseudomonas aeruginosa and enhanced the binding of sheep erythrocytes sensitized with IgG2b, when pre-incubated with macrophage monolayers for 30 min. Inhibition of both phagocytosis and binding of sensitized sheep erythrocytes was observed when tobramycin was co-incubated with the macrophage monolayers and indicator cells. Azlocillin, a semi-synthetic penicillin, caused inhibition of phagocytosis of opsonized bacteria and binding of sensitized sheep erythrocytes when added with the indicator cells. However, no effect was found if the macrophages were pre-incubated with azlocillin for 30 min. Ticarcillin, a semi-synthetic penicillin which is less active in vitro as an antimicrobial agent than azlocillin, had no effect on alveolar macrophages at the concentrations used in these assays. It is postulated that if these in vitro findings with rat alveolar macrophages are reflected in the in vivo situation in humans, the effect of antibiotics on host defence may be of clinical importance.

Animals↗

Antibiotic treatment of chronic Pseudomonas aeruginosa infection in cystic fibrosis patients.

The retrospective bacteriological results of 322 courses of anti-Pseudomonas aeruginosa chemotherapy in a cohort of 57 cystic fibrosis (CF) patients are reported. Tobramycin given as mono-therapy eradicated P. aeruginosa from the lungs of CF patients in 9% of the courses, whereas a combination therapy consisting of carbenicillin + tobramycin eradicated P. aeruginosa in 55% of the courses. However, the efficacy of the chemotherapy diminished successively when repeated courses of treatment were given. The efficacy of the carbenicillin + tobramycin combination in eradicating P. aeruginosa from the lungs of CF patients was compared with the efficacy of azlocillin + tobramycin and piperacillin + tobramycin in a prospective study. P. aeruginosa was eradicated in 78% of CF patients treated with carbenicillin + tobramycin, in 28% of CF patients treated with azlocillin + tobramycin, and in 33% of CF patients treated with piperacillin + tobramycin. However, the two latter groups of patients had on an average significantly higher numbers of P. aeruginosa precipitins in serum, indicating more severe infections. In CF patients where P. aeruginosa was not eradicated, a significant increase of MIC against carbenicillin, azlocillin and piperacillin was observed. There was a significant improvement of lung function and laboratory parameters reflecting diminished inflammation as a result of the treatment. 40% of the CF patients treated with azlocillin or piperacillin developed serum sickness-like symptoms.

Adolescent↗

In vitro activity and in vivo evaluation of ticarcillin plus clavulanic acid against aerobic and anaerobic bacteria.

The efficacy of ticarcillin plus clavulanic acid was compared with that of certain broad-spectrum antibiotics such as ticarcillin, azlocillin, and piperacillin against blood culture isolates of aerobic bacteria obtained from seriously ill patients and anaerobic bacteria obtained from other miscellaneous infections. Ticarcillin plus clavulanic acid was found to be as effective as other broad-spectrum antibiotics against most of the 285 septicemic isolates tested. Ticarcillin plus clavulanic acid was most effective against 351 anaerobic bacteria, including B. fragilis. Further, 32 strains of B. fragilis that were relatively resistant to ticarcillin and azlocillin were tested with a mixture of ticarcillin or azlocillin, each in combination with clavulanic acid. Ticarcillin plus clavulanic acid inhibited all 32 strains of B. fragilis. Addition of clavulanic acid to cephalothin, penicillin, or azlocillin also augmented the antibiotic activity against B. fragilis by 4- to 64-fold. These in vitro data suggest that ticarcillin plus clavulanic acid may be used as a single antibiotic in the cases of bacterial septicemias and that the combination may be used in the treatment of multiple-antibiotic-resistant bacterial strains. In a related study, the augmentation activity of clavulanic acid with penicillin or ticarcillin was evaluated against B. fragilis in a rat intra-abdominal abscess model. Gelatin capsules filled with a mixture of B. fragilis and Escherichia coli were implanted intraperitoneally in male Wistar rats. Four different groups of animals with appropriate controls were treated with penicillin or ticarcillin alone or in combination with clavulanic acid. Treatment was started immediately or delayed for 48 hours after peritoneal soilage. The mortality rate decreased by almost one half when antibiotic therapy was started immediately. Treatment with ticarcillin plus clavulanic acid resulted in a cure in 70 to 89 percent of animals, showing that this combination is the most effective regimen in the treatment of rats with experimental intra-abdominal abscesses caused by B. fragilis and E. coli.

Abscess↗

Mechanisms of resistance to beta-lactam antibiotics amongst Pseudomonas aeruginosa isolates collected in the UK in 1993.

Antimicrobial resistance among 1991 Pseudomonas aeruginosa isolates collected at 24 UK hospitals during late 1993 was surveyed. Three-hundred and seventy-two of the isolates were resistant, or had reduced susceptibility, to some or all of azlocillin, carbenicillin, ceftazidime, imipenem and meropenem, and the mechanisms underlying their behaviour were examined. Only 13 isolates produced secondary beta-lactamases: six possessed PSE-1 or PSE-4 enzymes and seven had novel OXA enzyme types. Those with PSE types were highly resistant to azlocillin and carbenicillin whereas those with OXA enzymes were less resistant to these penicillins. Chromosomal beta-lactamase derepression was demonstrated in 54 isolates, most of which were resistant to ceftazidime and azlocillin although susceptible to carbenicillin and carbapenems. beta-Lactamase-independent "intrinsic" resistance occurred in 277 isolates and is believed to reflect some combination of impermeability and efflux. Two forms were seen: the classical type, present in 195 isolates, gave carbenicillin resistance (MIC > 128 mg/L) and reduced susceptibility to ciprofloxacin and to all beta-lactam agents except imipenem; a novel variant, seen in 82 isolates, affected only azlocillin, ceftazidime and, to a small extent, meropenem. Resistance to imipenem was largely dissociated from that to other beta-lactam agents, and probably reflected loss of D2 porin, whereas resistance to meropenem was mostly associated with intrinsic resistance to penicillins and cephalosporins. Comparison of the present results with those of a similar study in 1982 revealed significant increases in the proportions of isolates with intrinsic resistance or stable derepression (p < 0.01, chi 2 test).(ABSTRACT TRUNCATED AT 250 WORDS)

Carbapenems↗

[Comparative activity of new semisynthetic penicillins and their combinations with gentamycin against gram-negative bacteria].

Comparative antibacterial activity of two novel ureidopenicillins (azlocillin and piperacillin), carbenicillin and ampicillin against 170 clinical strains of Enterobacteriaceae and 43 strains of Pseudomonadaceae was studied. Higher antibacterial activity of azlocillin and piperacillin evident from lower frequency of resistant strains and lower MICs for the majority of the isolates was shown. Impact of the inoculum size on the MIC values was observed with respect to all the penicillins. The study on the kinetics of Pseudomonadaceae death under the effect of azlocillin and carbenicillin revealed an increase in the bacteria growth after 6- to 8-hour contact with therapeutic concentrations of azlocillin and 4-hour contact with carbenicillin. Nor renewal of the culture growth was observed within 10-hour contact with combinations of the penicillins and 2 micrograms/ml of gentamicin.

Dose-Response Relationship, Drug↗

Overview of acylureidopenicillin pharmacokinetics.

The acylureidopenicillins which have been in man are azlocillin, mezlocillin, piperacillin, and furazlocillin (Bay k 4999). They exhibit dose dependent pharmacokinetics and accordingly upon increasing the doses have serum levels which are higher than the multiple of the dose, longer serum half-life (t1/2), and lower clearances (total serum clearance, renal clearance, and non-renal clearance). With doses of 1-2 g, t1/2 very between 0.7-1.1 h, and with 5.0 g 1.2-1.8 g. The elimination phase distribution volume corresponds to 10-30% of the body weight. The agents are excreted mainly through the kidneys. Referenced to the antibacterial activity of unchanged drugs, 50-80% of intravenous doses are eliminated in the urine. Only 25-35% of the dose of furazlocillin is excreted unchanged in the urine. The t1/2 is increased in reduced renal function, but mezlocillin is relatively little influenced by renal failure. With identical dose sizes, azlocillin appears to be subject to dose dependent pharmacokinetics to a higher degree than mezlocillin and piperacillin. Higher serum levels are also reached by azlocillin and the t1/2 of this agent is increased more in reduced renal failure than is the case for mezlocillin. The biliary levels of the acylureidopenicillins are high. A considerable biliary excretion occurs in reduced hepatic parenchymal function. The serum protein binding of these compounds decreases with higher concentrations varying between some 30% for 200 micrograms/ml and 50% for 2 micrograms/ml of azlocillin and mezlocillin, a mean of 16% for piperacillin in concentrations ranging from 20-300 micrograms/ml, and an average of 60% for furazlocillin. The acylureidopenicillins penetrate into tissues, cerebrospinal fluid and foetuses to produce therapeutic levels. The levels are rather low in bone tissue.

Absorption↗

Facilitated detection of antibiotic-resistant Pseudomonas in cystic fibrosis sputum using homogenized specimens and antibiotic-containing media.

Sputa from 30 patients with cystic fibrosis (CF) were cultured on routine and selective media plus three Mueller-Hinton antibiotic resistance screening plates containing tobramycin (5 micrograms/ml), azlocillin (100 micrograms/ml), and ticarcillin (100 micrograms/ml). In addition to direct semiquantitative plating, samples were homogenized for semiquantitative and quantitative culture. Blood agar plates from direct semiquantitative and homogenized semiquantitative cultures were then replica plated onto the antibiotic screening plates. Homogenized semiquantitative and quantitative cultures both detected more Pseudomonas aeruginosa strains than direct semiquantitative plating (103 versus 85 strains), including more antibiotic-resistant strains. Antibiotic screening media facilitated isolation of resistant strains and decreased detection time by 24 hr. Of the 103 strains on homogenized semiquantitative and quantitative cultures isolated before replica plating, 13 (13%) were tobramycin-resistant, 67 (65%) were ticarcillin-resistant, and 42 (41%) were azlocillin-resistant; 13 of 30 cultures (43%) had at least one tobramycin-resistant organism before replica plating. Replica plating detected an additional seven tobramycin-resistant and nine ticarcillin- or azlocillin-resistant strains in seven patients. Homogenization, antibiotic screening media, and replica plating enhance recognition of antibiotic-resistant strains in CF sputum.

Anti-Bacterial Agents↗

Once-daily versus multiple-daily gentamicin in empirical antibiotic therapy of febrile neutropenia following intensive chemotherapy.

The clinical efficacy and toxicity of once-daily compared with multiple-daily gentamicin dosing, in combination with azlocillin, were studied retrospectively in febrile neutropenic episodes following intensive chemotherapy. Fifty-two episodes were studied in 28 patients with acute myeloid leukaemia. Reasons for initiation of antibiotic therapy, dose, duration of treatment, organism isolation rates, response, cost comparison and toxicity were studied in the two treatment groups. The main indication for initiation of antibiotic therapy was neutropenic fever without a documented infection (80.8% of episodes). The response rate to once-daily gentamicin dosing and azlocillin was three times higher than to multiple-daily gentamicin dosing and azlocillin (P = 0.0112). The incidence of toxicity was low overall and was slightly but not significantly higher in the once-daily group. In this clinical context once-daily gentamicin at a dose of 7 mg/kg/day is more effective than a multiple-daily dosing regimen but may be more toxic.

Adult↗

Prospective randomized comparison of three antibiotic regimens for empirical therapy of suspected bacteremic infection in febrile granulocytopenic patients.

The standard regimen used by members of the European Organization for Research on Treatment of Cancer Antimicrobial Therapy Cooperative Group for empiric therapy of febrile neutropenic cancer patients has been treatment with ticarcillin plus amikacin. A three-arm prospective randomized controlled trial was performed to determine whether the extended-spectrum antipseudomonal penicillin azlocillin or the extended-spectrum cephalosporin cefotaxime had more or less efficacy than the beta-lactam in the ticarcillin-plus-amikacin regimen. A total of 742 patients from 22 institutions were evaluated. Single gram-negative rod bacteremias accounted for 83 episodes, and it was among these patients that the prognosis was least satisfactory, leading to a more intensive evaluation of this patient group. In these patients the azlocillin-plus-amikacin regimen resulted in a 66% response rate, compared with a 37% response rate for patients who received cefotaxime plus amikacin (P = 0.080) and a 47% response rate for patients who received ticarcillin plus amikacin (P = 0.207). The patients with gram-negative rod bacteremias and persistently profound granulocytopenia had substantially poorer response rates (37%) than the patients with rising granulocyte counts (73%; P = 0.004). A logistic regression analysis indicated that the following factors also affected infection resolution: beta-lactam utilization in the regimen (azlocillin was better than ticarcillin or cefotaxime), resolution of profound granulocytopenia (less than 100 cells per microliter) during therapy, and susceptibility to the beta-lactam antibiotic.

Adolescent↗

In vitro assessment of combined antibiotic and mucolytic treatment for Pseudomonas aeruginosa infection in cystic fibrosis.

The minimal inhibitory concentration of azlocillin for Pseudomonas aeruginosa is appreciably reduced when combined with the mucolytic agent mesna (Mistabron) because of an independent bacteriostatic effect of mesna. Bactericidal activity of azlocillin is unaltered by mesna. Mesna inhalations alone or combined with azlocillin may benefit cystic fibrosis patients with pseudomonas lung infections.

Azlocillin↗