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Biomedical subjects

J L Staneck

Publications and source records attributed to J L Staneck.

At least 19 recordsLinked to original sources

Dose effects of LPS on neutrophils in a whole blood flow cytometric assay of phagocytosis and oxidative burst.

Whole blood phagocytosis (P) and oxidative burst (OB), a rapid and sensitive flow cytometric method for quantifying neutrophil activation, was modified for single laser systems by using propidium iodide labeled Staphylococcus aureus and 2',7' dichlorofluorescein diacetate. The purpose of the present study was to characterize this assay with respect to the stimulatory activity of bacterial lipopolysaccharide (LPS) on phagocytosis. Blood from healthy donors was preincubated with log doses of bacterial LPS B (0.1-1,000 ng/ml) or sterile pyrogen-free saline at 37 degrees C from 0-120 minutes. LPS increased both P and OB in a dose-dependent manner (up to 62 and 121%, respectively) at all time points tested, and this effect on P and OB could be detected even with no preincubation. This LPS-induced phagocytic activity could be blocked by the addition of polymyxin B (10 micrograms/ml) during preincubation. The priming effect of LPS was maximal at 45 min. P and OB were inhibited by preincubation with EDTA at doses greater than 2 mM (60 and 80% inhibition, respectively). These observations are consistent with the exquisite sensitivity of the neutrophil to endotoxin. This method can evaluate neutrophil response to immunomodulatory and chemotherapeutic agents in a physiological milieu. These findings re-emphasize the necessity of using pyrogen-free reagents in any study of neutrophil function.

Anticoagulants

Niacin-positive Mycobacterium kansasii isolated from immunocompromised patients.

Niacin-positive Mycobacterium kansasii was isolated from three patients, two with respiratory infections and one with a perirectal abscess. The isolates were phenotypically similar to other strains of M. kansasii, differing only in their ability to produce niacin. This phenotype has been reported only twice in the literature, during the 1960s.

Adult

Survival of Streptococcus pneumoniae in corneal storage media.

The Cincinnati Eye Bank had six corneoscleral rims in which Streptococcus pneumoniae was cultured after preservation in corneal storage media. To determine the survival of this organism under conditions common for corneal storage, gentamicin-supplemented McCarey-Kaufman (M-K) medium and chondroitin sulfate/Dextran medium (Dexsol, Ciron Ophthalmics, Irvine, CA, U.S.A.) were inoculated with S. pneumoniae and kept at 4 degrees C. Thioglycollate broth plus 10% rabbit serum (Thio-S) and tryptic soy broth (TSB) served as growth controls. At day 14 after inoculation of 10(5) colony-forming units (CFU)/ml, Dexsol showed a 1-log decrease in bacterial concentration, the M-K medium a 2-log decrease and Thio-S a 4-log decrease, whereas TSB showed no detectable organisms. By day 21 Dexsol had only a 2-log decrease in bacteria. These data suggest that corneal storage medium supplemented with gentamicin does not exert bactericidal activity against S. pneumoniae and may actually support its survival at 4 degrees C.

Colony Count, Microbial

Reemergence of epidemic methicillin-resistant Staphylococcus aureus in a general hospital associated with changing staphylococcal strains.

An epidemic of methicillin-resistant Staphylococcus aureus (MRSA) infections involving 323 patients occurred at the University of Cincinnati Hospital from 1977 to 1981. Subsequently, endemic MRSA persisted in the hospital for 6 years, until 1987, when a new epidemic began with 223 patients becoming infected over 3 years. Between the two epidemics, there was a major change in the MRSA recovered from infected patients, as demonstrated by three epidemiologic markers. Antibiograms showed that the tetracycline-resistant MRSA involved in the first epidemic was replaced by tetracycline-susceptible MRSA in the second epidemic; bacteriophage typing indicated that the original epidemic strain, D11/83A/85, had been replaced by new strains, many of which were susceptible to phage 54; and restriction endonuclease analysis of plasmid DNA confirmed that a single strain was involved in the first epidemic and that multiple strains were present in the second epidemic. The epidemiology of MRSA infections in the hospital changed with the change in staphylococcal strains. The first epidemic was hospital based with most infections occurring in surgical patients, and the burn unit was the major reservoir. In contrast, 28% of the patients in the second epidemic had community-acquired infections, and nursing home patients were an important source of these infections. Also, 29% of the hospital-acquired infections in this epidemic occurred in nonsurgical patients. This time the burn unit was not a reservoir of infection, but other intensive care units were. The increased diversity of strains of MRSA in the second epidemic might be related to increased transmission in the community and more widespread transmission in the hospital.

Bacteriophage Typing

Wide variability in Pseudomonas aeruginosa aminoglycoside results among seven susceptibility testing procedures.

Seven commonly used antimicrobial susceptibility testing methods were used to test the susceptibility of 150 isolates of Pseudomonas aeruginosa against gentamicin, tobramycin, amikacin, carbenicillin, and piperacillin. Results were compared with respect to the susceptibility characteristics of the population of isolates as defined by each method. Conventional methods included agar disk diffusion and agar dilution, carried out in accordance with current recommendations of the National Committee for Clinical Laboratory Standards, as well as broth microdilution testing with cation-supplemented Mueller-Hinton broth (CSMHB). Methods in which instrumentation was used for result determination included the Autobac I, Avantage, Sensititre Autoreader (using a breakpoint panel at 18 h of incubation), and Vitek (AMS-240, using the GNS susceptibility card). When necessary for comparison, MIC data were converted to categorical interpretations (susceptible, intermediate, and resistant). With respect to gentamicin, no significant differences were noted among the results of disk diffusion, broth microdilution, Sensititre Auto breakpoint, or Vitek methods which characterized 60 to 67% of isolates as susceptible, 16 to 22% as intermediate, and 13 to 17% as resistant. In contrast, agar dilution, Autobac, and Avantage, although yielding gentamicin results similar to those of one another, were each significantly different in result reporting from the other four methods above for gentamicin results, and they characterized the Pseudomonas population largely as susceptible (88 to 97%), with 0 to 6% intermediate and only 3% to 6% resistant. More isolates were characterized as being resistant to gentamicin in the Avantage test if an assay broth supplemented with increased amounts of calcium was used. Cation impregnation of Autobac disks did not appreciably change Autobac results. The geometric mean MIC of gentamicin was 4 micrograms/ml lower in the agar dilution method than in the CSMHB microdilution method, despite monitoring of the agar for cation content through performance disk diffusion testing with P. aeruginosa ATCC 27853. Tobramycin activity was greater than gentamicin activity, and susceptibility to tobramycin ranged from 89 to 97%, with few statistically significant differences noted among the seven methods studied. Differences in MIC distribution and geometric mean MIC between agar dilution and CSMHB microdilution testing were minimal and suggested less of a cation influence on tobramycin than gentamicin results. Although amikacin was also more active than gentamicin (83 to 99% of isolates were susceptible), differences in the amikacin results among methods tended to reflect the same trends in reporting as seen with gentamicin testing, with the exception that results of Avantage testing were similar to those of disk diffusion, CSMHB microdilution, Sensititre, and Vitek. A difference in geometric mean MIC of 5 micrograms/ml between CSMHB testing and agar dilution testing suggested the influence of divalent cations on amikacin results. Few highly significant differences were noted among methods when isolates were tested against carbenicillin and piperacillin, except that Avantage piperacillin results (66% susceptible) and Autobac piperacillin results (98% susceptible) were noticeably different from the percent piperacillin susceptibility (range, 85 to 92%) measured by the other methods. Method-dependent variability among aminoglycoside susceptibility results, particularly when testing gentamicin, prevents meaningful comparison of Pseudomonas susceptibility trends among hospitals when different methods are used and promotes confusion and frustration among clinical microbiologists and clinicians owing to the uncertainties of clinical meaning of these data.

Aminoglycosides

Rapid MIC testing with the sensititre autoreader.

Automated microdilution MIC results, obtained with the Autoreader (Sensititre, Inc., Salem, N.H.) following 5 h of incubation, were compared with manually read, concurrent control MICs following 18 h of incubation in a three-laboratory comparative study. A total of 704 members of the family Enterobacteriaceae or similar gram-negative organisms were tested against 17 antimicrobial agents. Autoreader MICs were within 1 doubling dilution of control values in 92.9% of instances. Discrepancies of +/- 2 doubling dilutions and +/- 3 or greater doubling dilutions were noted in 4.5 and 2.6%, respectively, of the 7,687 drug-organism combinations analyzed. The majority of errors occurred when beta-lactam antimicrobial agents were tested with a variety of different species. MICs at 5 h, when Pseudomonas aeruginosa was used, were possible in only half the isolates tested and yielded data on only a limited number of drugs in the remaining instances. Excluding results obtained with penicillin and ampicillin, which were uniformly poor, Staphylococcus aureus Autoreader values were within +/- 1 doubling dilution of control values in 93.6% of instances, 5.4% varied by +/- 2 dilutions, and only 1% of test values by +/- 3 or more dilutions from control values for 82 isolates tested against nine antimicrobial agents. Of eight additional S. aureus isolates tested that were resistant to methicillin, only one was read correctly by the Autoreader, with results on the remaining seven appearing as either insufficient growth or as total resistance to all drugs tested. Interlaboratory reproducibility was excellent for selected isolates of S. aureus and gram-negative bacilli. The accuracy of the Sensititre Autoreader MIC results was comparable to that of other same-day quantitative systems for members of the family Enterobacteriaceae and S. aureus, while the economic and procedural advantages of the broth microdilution method was maintained.

Anti-Bacterial Agents

Legionella cincinnatiensis sp. nov. isolated from a patient with pneumonia.

A Legionella-like organism (strain 72-OH-H [= ATCC 43753]) was isolated from an open-lung biopsy specimen from a hemodialysis patient with end-stage renal disease and bronchopneumonia. Growth characteristics and gas-liquid chromatographic profiles of the isolate were consistent with those for Legionella spp. The isolate was presumptively identified as a Legionella longbeachae serogroup 1 strain by direct immunofluorescence staining. However, the organism was serologically distinct in the slide agglutination test with absorbed antisera. DNA hybridization studies showed that strain 72-OH-H constitutes a new Legionella species, which is named Legionella cincinnatiensis (ATCC 43753).

Adult

Microbiology of adult cellulitis.

Needle aspiration of cellulitis sites is commonly advocated to assist in the identification of causative organisms. Twenty-five nondiabetic, adult patients with a clinical diagnosis of cellulitis had site aspirations and blood cultures obtained before antibiotic therapy was initiated. Site cultures were positive in 6 of 25 patients. Blood cultures were positive in 4 of 25 patients. All organisms except one (Enterobacter agglomerans) were staphylococci or streptococci. The gram-negative bacilli were not believed to be a pathogen based on the patient's prompt response to nafcillin. In adult patients who do not have complications, the use of needle aspiration was not supported. Empiric treatment of cellulitis aimed at gram-positive cocci appears to be sufficient.

Adult

Bronchoalveolar lavage for diagnosing acute bacterial pneumonia.

The utility of gram stain and semiquantitative culture of the fluid retrieved by bronchoalveolar lavage (BAL) in identifying the causative agent in acute bacterial pneumonia was initially assessed in 92 patients. Fifteen of these patients presented with clinically active bacterial pneumonia; the remaining patients underwent bronchoscopy to evaluate other processes in the lung. Thirteen of the 15 patients with clinically active bacterial pneumonia had a BAL culture greater than or equal to 10(5) colony-forming units per milliliter of BAL fluid, whereas none of the other groups had a positive culture (chi 2 = 70.7, P less than .001). Gram stain of cytocentrifuged BAL fluid was positive (one or more organisms seen per 1,000 X field) only in those patients with an active bacterial pneumonia. Applying this technique, we studied 59 immunocompromised patients presenting with pulmonary infiltrates. Eight (21%) of the 39 patients presenting with microbial-related infiltrates proved to have acute bacterial pneumonia by BAL culture; the pneumonia resolved with appropriate antimicrobial therapy.

Adolescent

Clinical evaluation of efficacy, pharmacokinetics, and safety of teicoplanin for serious gram-positive infections.

Nineteen patients hospitalized for serious gram-positive infections were treated with teicoplanin, a new glycopeptide antibiotic. A variety of infections were treated, including endocarditis, septic thrombophlebitis, osteomyelitis, pyogenic arthritis, and soft tissue infection. Of 13 infections that could be evaluated in 12 patients, there were 8 clinical cures, 2 improvements, 1 recurrence, and 2 failures. Of the eight patients with Staphylococcus aureus bacteremia, seven were clinically cured or improved with teicoplanin therapy. Of the nine patients in whom the bacteriological response to treatment could be fully evaluated, six were cured; there was recurrence of infection in one, and treatment failed in two patients. In vitro testing showed the 13 bacterial isolates (9 S. aureus, 3 S. epidermidis, and 1 group B streptococcus) to be uniformly susceptible to teicoplanin, with MICs ranging from 0.12 to 0.5 microgram/ml. Every isolate was more susceptible in vitro to teicoplanin than to vancomycin. Three of the staphylococcal isolates were resistant to methicillin. Pharmacokinetic studies demonstrated that after an initial drug-accumulation period, a single daily dose adequately maintained the teicoplanin concentrations in serum within therapeutic ranges. Teicoplanin also penetrated well into synovial fluid. The drug was well tolerated by either intravenous or intramuscular administration. The most significant adverse reaction was an urticarial rash which required discontinuation of therapy in one patient; a second patient experienced a modest decrease in high-frequency auditory threshold. Asymptomatic eosinophilia and mild elevation of serum transaminases were noted as well. The results of this study suggest that teicoplanin is a safe and effective new agent for treatment of serious infections caused by gram-positive organisms.

Adult

Sensititre autoreader for same-day breakpoint broth microdilution susceptibility testing of members of the family Enterobacteriaceae.

The Sensititre Autoreader system is an instrument-assisted broth microdilution susceptibility test procedure based on the detection of fluorogenic growth substrate metabolism by test bacteria with different concentrations of antimicrobial agents. In the current investigation, this system was assessed as a means for predicting the in vitro activity of 17 antimicrobial agents versus numerous species of the family Enterobacteriaceae and Pseudomonas aeruginosa by using a breakpoint broth microdilution test format. Same-day and overnight determinations of susceptibility were made with the Sensititre Autoreader system, and in both cases, the results were compared with those obtained with a manual overnight breakpoint broth microdilution susceptibility test. Among a total of 6,086 organism-antimicrobial agent comparisons with Enterobacteriaceae, concordance was noted between the results of the same-day Autoreader system and the manual overnight test in 94.4% of cases. The same-day Autoreader results with members of the Enterobacteriaceae other than Proteus spp. were determined after 4 h of incubation; with Proteus spp. the same-day Autoreader results were determined after 5 h of incubation. When the Enterobacteriaceae Autoreader results were determined after 18 h of incubation, concordance was noted in 97.2% of comparisons. Among a total of 1,377 organism-antimicrobial agent comparisons with P. aeruginosa after 18 h of incubation, agreement of results from the manual overnight test and the Autoreader system was achieved in 92.2% of cases.

Anti-Bacterial Agents

Performance characteristics of a commercially prepared biphasic blood culture bottle.

A biphasic blood culture bottle (BiPB: GIBCO Laboratories, North Andover, Mass.) with an architectural design that physically separates the agar slant from the broth was compared with a conventional vented monophasic bottle (MPB-A) for use in the routine culture of blood. Both bottles contained tryptic soy broth. Tryptic soy agar was used for the BiPB slant. A third unvented bottle (MPB-N) with Columbia broth was included as part of the blood culture set. Of 3,537 sets collected, 444 were positive; 57 of these 444 sets were positive by virtue of an exclusively positive anaerobic bottle. Both BiPB and MPB-A were positive in 235 of the remaining 387 positive sets. A total of 521 isolates was recovered during the study. Of these isolates, 252 were recovered in both the BiPB and the MPB-A from the same set; 105 isolates grew in the BIPB but not in MPB-A, 95 isolates grew only in the MPB-A but not in BiPB, and 69 grew exclusively in the MPB-N. The BiPB allowed more rapid recovery of Candida spp., J-K diphtheroids, Pseudomonas spp. Making BiPB subcultures was easy enough to permit both early and daily subculture, which provided isolated colonies sooner than could be done by using the MPB-A. Isolated colonies and, therefore, identification and susceptibility results were available at least 1 day earlier for the BiPB isolates in approximately 50% of instances when both the BiPB and the MPB-A were positive. Staphylococcus epidermidis and streptococci were recovered more frequently in the BiPB, while gram-positive anaerobes were detected at a significantly (P less than 0.025) more frequent rate in the MPB-A than in the BiPB. Either bottle, however, should be used in conjunction with an anaerobic bottle for optimal recovery of anaerobic bacteria.

Bacteria

Imipenem susceptibility testing with a commercially prepared dry-format microdilution tray.

MICs of imipenem, concurrently generated in commercially prepared microdilution trays containing predried antibiotic dilutions (Sensititre), and in a standard agar dilution assay (as recommended by the National Committee for Clinical Laboratory Standards, Villanova, Pa.), were within +/- 1 twofold dilution for 94% of 226 bacterial isolates. Imipenem biological activity remained stable over 5 months of tray storage at room temperature against Pseudomonas aeruginosa ATCC 27853. Activity of imipenem was shown by microdilution testing with 890 clinical isolates to be high, with only 4% of isolates having MICs of greater than or equal to 16 micrograms/ml (in vitro resistance).

Bacteria

Screening tests and "rapid" identification. Is anybody out there listening?

The responses by 55 of 300 physicians queried at the University Hospital, Cincinnati, to an extensive questionnaire, form the basis for this comment on the clinical impact of screening tests and rapid identification. Examination and interpretation of smears of fluids and tissue were felt to provide useful data within a short time of specimen collection and the majority of physician respondents were willing to sacrifice some degree of accuracy in results in return for an answer delivered shortly after specimen submission. Screening for bacteriuria was felt to be useful if the test was sensitive enough to rule out infection as suggested by the presence of approximately 10,000 CFU/ml of urine, if results could be had within 1 hr of specimen collection for outpatients, 2 hr for inpatients, and if test results for pyuria accompanied the screen results. The value of taxonomy, in general, was directly related to how much knowing the taxon would direct antibiotic therapy. The time required for a taxonomic result seemed to matter only when susceptibility test results were either delayed or not forthcoming. Interest in the taxonomy of anaerobic bacteria appeared to be limited to a small group of physician respondents (infectious disease specialists) and also was relative to the ability of the taxonomic designation to evoke a specific therapeutic regimen. In summary, the impact of microbiologic data on the practice of medicine appears to depend on the ability of the laboratory to complete the entire test process, from order to transmission of answer, within a time frame perceived to be useful by the clinician.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriological Techniques

Prolonged hospital epidemic of salmonellosis: use of trimethoprim-sulfamethoxazole for control.

The occurrence of a foodborne outbreak of Salmonella drypool, an uncommon serotype, led to the recognition of hospital-acquired cases occurring over a 5-year period. In late 1981, a catered luncheon resulted in an epidemic of S. drypool, and in retrospect, cases were found to have been occurring in hospital patients from 1977 until 1981. Over the next 4 months, patients, employees, and 50% of the kitchen workers became infected, despite repeated culture surveys of the staff and removal of all infected workers. Food and environmental cultures failed to reveal the source of infection, but infected workers were observed to have multiple negative cultures over several months and then to become positive again. Closing of the kitchen for 2 weeks failed to stop the epidemic. Finally, all kitchen employees, both infected and uninfected, were treated with trimethoprim-sulfamethoxazole for 2 weeks. After treatment, no new kitchen workers became infected, although two workers who had had positive cultures earlier in the epidemic began to excrete S. drypool again, and the epidemic ended.

Carrier State

Clinical trial of the efficacy and safety of ticarcillin and clavulanic acid.

Forty-three hospitalized patients were treated with a new antibiotic combination containing ticarcillin plus the beta-lactamase inhibitor, clavulanic acid, in a fixed combination for intravenous use. A variety of infections were treated, including pneumonia, bacteremia, urinary tract infection, and osteomyelitis. Of 50 episodes of infection in 43 patients, 44 clinical cures were obtained, with 5 patients improving and 1 patient failing to respond to treatment. In vitro susceptibility testing of 101 clinical isolates was notable for the rarity of resistance to the combination antibiotic. Of specific interest, all 14 isolates of Staphylococcus aureus were susceptible to ticarcillin plus clavulanic acid, whereas only 2 of the 14 isolates were susceptible to ticarcillin alone. Adverse reactions to the study drug were minimal; eosinophilia, unaccompanied by other allergic phenomena, and oral candidiasis were most frequent. Overall, the combination of ticarcillin with the beta-lactamase inhibitor, clavulanic acid, appears to be a safe and effective drug for the treatment of infections caused by susceptible organisms.

Aged