Results of the c-TRAK TN trial: a clinical trial utilising ctDNA mutation tracking to detect molecular residual disease and trigger intervention in patients with moderate- and high-risk early-stage triple-negative breast cancer.

BACKGROUND: Post-treatment detection of circulating tumour DNA (ctDNA) in early-stage triple-negative breast cancer (TNBC) patients predicts high risk of relapse. c-TRAK TN assessed the utility of prospective ctDNA surveillance in TNBC and the activity of pembrolizumab in patients with ctDNA detected [ctDNA positive (ctDNA+)]. PATIENTS AND METHODS: c-TRAK TN, a multicentre phase II trial, with integrated prospective ctDNA surveillance by digital PCR, enrolled patients with early-stage TNBC and residual disease following neoadjuvant chemotherapy, or stage II/III with adjuvant chemotherapy. ctDNA surveillance comprised three-monthly blood sampling to 12 months (18 months if samples were missed due to coronavirus disease), and ctDNA+ patients were randomised 2 : 1 to intervention : observation. ctDNA results were blinded unless patients were allocated to intervention, when staging scans were done and those free of recurrence were offered pembrolizumab. A protocol amendment (16 September 2020) closed the observation group; all subsequent ctDNA+ patients were allocated to intervention. Co-primary endpoints were (i) ctDNA detection rate and (ii) sustained ctDNA clearance rate on pembrolizumab (NCT03145961). RESULTS: Two hundred and eight patients registered between 30 January 2018 and 06 December 2019, 185 had tumour sequenced, 171 (92.4%) had trackable mutations, and 161 entered ctDNA surveillance. Rate of ctDNA detection by 12 months was 27.3% (44/161, 95% confidence interval 20.6% to 34.9%). Seven patients relapsed without prior ctDNA detection. Forty-five patients entered the therapeutic component (intervention n = 31; observation n = 14; one observation patient was re-allocated to intervention following protocol amendment). Of patients allocated to intervention, 72% (23/32) had metastases on staging at the time of ctDNA+, and 4 patients declined pembrolizumab. Of the five patients who commenced pembrolizumab, none achieved sustained ctDNA clearance. CONCLUSIONS: c-TRAK TN is the first prospective study to assess whether ctDNA assays have clinical utility in guiding therapy in TNBC. Patients had a high rate of metastatic disease on ctDNA detection. Findings have implications for future trial design, emphasising the importance of commencing ctDNA testing early, with more sensitive and/or frequent ctDNA testing regimes.

Cutaneous presentation of progressive disseminated histoplasmosis nine years after renal transplantation.

Initial presentation of invasive fungal infections such as histoplasmosis can include non-specific clinical manifestations, especially in immunocompromised patients. A high index of suspicion is required to identify atypical manifestations of these diseases, which carry a high risk of mortality, if the diagnosis is delayed or missed. We describe a case of a kidney transplant recipient with cutaneous lesions as initial manifestation of progressive disseminated histoplasmosis where a skin biopsy was crucial to an early diagnosis.

Diabetic foot infections. Bacteriologic analysis.

Diabetic patients with foot infections were prospectively evaluated over a two-year period. Cultures from reliable specimens avoiding contamination with foot ulcers were obtained in 54 infectious episodes. Staphylococcus species, Enterococcus species, Corynebacterium species, and various species of Enterobacteriaceae were commonly isolated. Common anaerobic isolates included Peptostreptococcus magnus, Peptostreptococcus prevotii, and Bacteroides species. Results of cultures from 94 unreliable specimens were similar. Results of reliable and unreliable specimens obtained simultaneously in 26 patients agreed in seven (27%), but antibiotics selected for organisms isolated from unreliable specimens would have adequately covered pathogens found in the reliable culture in 24 (93%). Diabetic foot infections usually involve mixed bacterial flora, including aerobic, facultatively anaerobic, and anaerobic microorganisms. Specimens should be obtained from infected tissue that does not communicate directly with the foot ulcer if possible. If such specimens are not available, cultures of purulent exudate within the foot ulcer or soft-tissue sinuses may provide useful information on which to base decisions about antibiotic therapy. Broad-spectrum beta-lactam antibiotics or a combination of antibiotics active against facultatively anaerobic cocci and bacilli as well as anaerobes provide the best empirical antimicrobial coverage in these patients.

Infection prevention in severely myelosuppressed patients: a comparison between ciprofloxacin and a regimen of selective antibiotic modulation of the intestinal flora.

PURPOSE: To study whether oral ciprofloxacin would be as effective in preventing bacterial infections in severely myelosuppressed patients as selective antibiotic modulation of the gut flora with neomycin/polymyxin B sulfate/nalidixic acid (NPN). PATIENTS AND METHODS: One hundred and five patients undergoing allogeneic or autologous bone marrow transplant, or induction therapy for acute leukemia in 1988 and 1989 were studied. Patients were stratified according to the type of therapy, and randomized in a ratio of 2:1 to either oral ciprofloxacin 500 mg BID, or a combination of oral neomycin 250 mg QID, polymyxin-B 100 mg QID, and oral nalidixic acid 1,000 mg BID. Treatment began on admission and continued until the absolute granulocyte count was greater than 500/mm3 for 3 consecutive days. RESULTS: The 96 evaluable patients were evenly distributed over the 3 treatment groups; 63 patients received ciprofloxacin and 33 received NPN. Fever developed in 92% of patients on ciprofloxacin and in 97% of patients on NPN. (P = 0.66), 6.6 +/- 5.8 and 7.2 +/- 5.3 days from the start of prophylaxis, respectively. Twenty-five patients on ciprofloxacin developed 29 microbiologically documented infections, fewer than the 26 infections in the 22 patients on NPN (P = 0.02). Patients on ciprofloxacin had fewer bacteremias (33%) than did the NPN patients (55%) (P = 0.05). Gram-negative bacteremias were very rare (2 cases; no Enterobacteriaceae), but streptococcal bacteremias were frequent in both arms (27 cases). Side effects were not significantly different, but compliance with ciprofloxacin was better. CONCLUSIONS: Ciprofloxacin is at least as effective as the combination of neomycin/polymyxin/nalidixic acid in the prophylaxis of bacterial infections in myelosuppressed patients, and is better tolerated. Additional agents to prevent streptococcal infections are needed.

Studies on antifungal agents. 23. Novel substituted 3,5-diphenyl-3-(1H-imidazol-1-ylmethyl)- 2-alkylisoxazolidine derivatives.

The synthesis and antifungal activity of a novel series of substituted 3,5-diphenyl-3-(1H-imidazol-1-ylmethyl)-2-alkylisoxazolidine derivatives (15-30) are described. The synthesis of the title compounds was accomplished via a 1,3-dipolar cycloaddition reaction of alpha-substituted ketonitrones with appropriate styrene precursors. The compounds when tested in vitro in solid agar cultures exerted a very potent antifungal activity against a wide variety of yeast and systemic mycoses and dermatophytes, especially Trichophyton and Microsporum sp., Epidermophyton floccosum and Candida stellatoidea. The in vitro activity against Aspergillus fumigatus and Candida albicans was moderate to potent. Overall, the two bis(4-chlorophenyl) analogues 18 and 19 were the most potent in vitro compounds, showing MIC values ranging between 0.2 and 7.0 microgram/mL, as compared to 0.2-20.0 micrograms/mL for ketoconazole, which was used as the positive standard in all assays. When tested in vivo in the rat vaginal candidiasis model, derivative 18, although showing significant antifungal activity when compared to controls, was less effective than ketoconazole. The title 3,5-substituted isoxazolidine compounds represent a novel class of potent antifungal agents.

Anaerobic flora of the normal human conjunctival sac.

Specimens from the conjunctival sacs of 92 healthy eyes were cultured on two separate occasions to determine the presence or absence of a persistent anaerobic flora. Aerobic bacteria and fungi were also studied for comparison. Of the 184 eye cultures, 112 (60.9%) contained at least one microorganism. Obligate anaerobes were recovered from 51.6% (95/184) of the cultures. Propionibacterium acnes, the predominant anaerobe encountered, was present in 49.5% (91/184) of the eyes. Aerobic and facultatively anaerobic bacteria, present in 32.6% of the total eye cultures, were less common than obligate anaerobes. Staphylococcus epidermidis was the most common of these. Fungi were rarely found. Our findings suggest that the conjunctival sac is either sterile or normally contains small numbers of anaerobic, aerobic, or facultatively anaerobic bacteria. As other workers have suggested, the origin of the bacteria in the eyes may be the skin of the eyelids.

Clinical, pathogenetic, and laboratory features of Capnocytophaga infections.

Granulocytopenia and oral mucosal defects have been reported to be important predisposing factors to recently recognized cases of Capnocytophaga septicemia. The authors call attention to an apparent preponderance of these cases in the pediatric age group and emphasize laboratory features which they have found helpful in the diagnosis of Capnocytophaga infections. Thirteen patients with Capnocytophaga infections were seen during a seven-year period. Seven of these patients had Capnocytophaga bacteremia. Six of seven bacteremic patients were granulocytopenic, six had oral mucosal defects, and three died. Five of the seven bacteremic patients were younger than 20 years of age. This represents a disproportionate distribution of cases in the pediatric age group within the author's institution, because 43% of blood culture specimens submitted to their microbiology laboratory are obtained from pediatric patients. This observation is supported by a review of the reported cases of Capnocytophaga septicemia in which 7 of 12 patients were younger than 20 years of age. Because Capnocytophaga may superficially resemble the more commonly isolated Fusobacterium nucleatum, distinguishing features for laboratory identification are discussed.

Development and evaluation of diagnostic radiometric assays for serogroup 4 L. pneumophila urinary antigens.

We previously reported that 80% of patients with serogroup 1 Legionella pneumophila pneumonia excrete detectable quantities of specific antigens in their urine. The purpose of this study was to determine whether specific antigens can be detected in urine from patients with serogroup 4 L. pneumophila pneumonia. Antisera were prepared in 15 rabbits and used to set up 25 solid-phase radioimmunoassays. The best of these assays detected specific antigens in urine from 4 of 5 patients with culture-proven serogroup 4 infections and from 1 of 2 patients with culture-proven serogroup 10 infections. None of 100 control specimens was positive in the assays. The antigens from serogroup 4 and 10 infections behaved differently in the assays, suggesting that they are not identical molecules. This study demonstrates that it is possible to detect specific antigens in the urine of patients with serogroup 4 L. pneumophila pneumonia. This may be a useful method for diagnosing these infections rapidly.

Multicenter comparison of the high volume (10 ml) NR BACTEC PLUS system and the standard (5 ml) NR BACTEC system.

This multicenter study was designed to compare the new BACTEC PLUS system (nonradiometric), which utilizes an 8- to 10-ml blood inoculum in a resin-containing medium, to the standard BACTEC (nonradiometric) without resins and 5-ml blood inoculum. There were 12,341 compliant sets studied, yielding 1331 positives, with 1099 sets deemed clinically significant. Overall the BACTEC PLUS showed an enhanced recovery of 33% (p less than 0.001) over its standard counterpart, with significant yield increased in the staphylococci (p less than 0.001), streptococci (p less than 0.002), pseudomonads (p less than 0.002), Enterobacteriaceae (p less than 0.001), and other aerobic Gram negatives (p less than 0.02). The enhanced performance increased to 53% if the patient was receiving any antibiotics at the time the blood was cultured. In patients known to be free of antibiotics at the time of blood draw, there was still an increased yield of 18%. The new system detected positivity at least one reading sooner than twice as often as the converse, and confirmed septic episodes significantly more often (21% overall) (41% on antibiotics) (15% no antibiotics). The BACTEC PLUS has distinct advantages over its low blood volume, nonresin counterpart.

In vitro comparison of E4868, a new trifluoroquinolone, with ciprofloxacin and temafloxacin tested against 5192 recent clinical isolates from five medical centers.

The in vitro activity of E4868 was compared with that of ciprofloxacin and temafloxacin in a multicenter study. More than 90% of Enterobacteriaceae and 82% of the 651 isolates of Pseudomonas aeruginosa, as well as > 95% of the oxacillin-susceptible staphylococci, 100% of the beta-hemolytic streptococci, and 76% of the enterococcal isolates were susceptible to E4868 (minimum inhibitory concentrations, < or = 2 micrograms/ml). E4868 was generally equivalent or slightly less active than ciprofloxacin against the Enterobacteriaceae and Pseudomonas and more active against staphylococci and beta-hemolytic streptococci. The activity of E4868 was generally equivalent to or slightly greater than that of temafloxacin against all major groups of organisms.

Ceftizoxime and cefoxitin susceptibility testing against anaerobic bacteria: comparison of results from three NCCLS methods and quality control recommendations for the reference agar dilution procedure.

Ceftizoxime and cefoxitin (control) were tested against 99 anaerobic bacteria using three commonly used susceptibility testing methods. The cefoxitin MIC results with the National Committee for Clinical Laboratory Standards (NCCLS) reference agar dilution (RAD) method and the broth microdilution (BMD) method did not differ by more than one log2 dilution step. However, the ceftizoxime BMD MIC results were 2- to 4-fold lower than those produced by the RAD procedure. Broth disk elution (BDE) tests for both cephalosporins (ceftizoxime and cefoxitin) had very high rates of false-susceptible and false-resistant error when compared to the RAD MICs: the BDE tests are not recommended for either cephalosporin. Findings indicate that because of method and possible medium or technical variability, ceftizoxime MICs should be determined only by the NCCLS RAD method. Also, cefoxitin susceptibility should be assessed primarily by RAD or BMD procedures. This should minimize confusion related to ceftizoxime's spectrum and activity against anaerobic bacteria compared to similar beta-lactam drugs. Ceftizoxime quality control guidelines were established for Bacteroides fragilis (ATCC 25285): mode, 64 micrograms/ml, range 32-128 micrograms/ml. Other control strains were not reliable.

Piperacillin/tazobactam (YTR 830) combination. Comparative antimicrobial activity against 5889 recent aerobic clinical isolates and 60 Bacteroides fragilis group strains.

Piperacillin combined with tazobactam (formerly YTR 830) was tested at a ratio of 8:1 against 5889 aerobic isolates and 50 strains from the Bacteroides fragilis group. Imipenem was the most active agent tested against Enterobacteriaceae (99.3% at less than or equal to 4 micrograms/ml), ceftazidime was most effective against nonenteric Gram-negative bacilli (80.7% at less than or equal to 8 micrograms/ml), and piperacillin/tazobactam possessed a superior spectrum against Gram-positive cocci (92.2% at less than or equal to 16/2 micrograms/ml). Against all aerobic strains, piperacillin/tazobactam had a spectrum (90.3% at less than or equal to 16/2 micrograms/ml) comparable to imipenem (93.6% at less than or equal to 4 micrograms/ml) and was distinctly greater than that of ticarcillin/clavulanic acid (73.3% at less than or equal to 16/2 micrograms/ml) and ceftazidime (75.5% at less than or equal to 8 micrograms/ml). Against the B. fragilis group isolates, all piperacillin/tazobactam MICs were less than or equal to 64/8 micrograms/ml. This activity was superior to piperacillin alone (MIC:50, 8-64 micrograms/ml) and cefoxitin (MIC50, 4-64 micrograms/ml). Piperacillin/tazobactam appears to be a promising parenteral antimicrobial combination, with a spectrum effective against a wide variety of clinical pathogens.

Effect of clavulanic acid on the susceptibility of clinical anaerobic bacteria to ticarcillin: a multicenter study.

Over 2,000 recent clinical isolates of anaerobic bacteria were tested at five medical centers for susceptibility to ticarcillin, ticarcillin plus clavulanic acid, clindamycin, and metronidazole. At 64 micrograms/ml, ticarcillin inhibited 92% of all isolates, but 98% were inhibited at this concentration when 2 micrograms/ml of clavulanic acid was added. With different Bacteroides species, clavulanic acid reduced ticarcillin MICs 2- to 32-fold; other anaerobic species were not significantly affected.

Antimicrobial activity of U-76,252 (CS-807), a new orally administered cephalosporin ester, including recommendations for MIC quality control.

U-76,253A (R-3746), the active metabolite of the new cephalosporin ester, U-76,252 (CS-807), was tested against 4,742 fresh clinical isolates from four large medical centers. U-76,253A was very active against nearly all species of Enterobacteriaceae (87.7% inhibited at less than or equal to 4.0 micrograms/ml). Staphylococcus spp., and the streptococci. The U-76,253A spectrum was superior to the comparison orally administered cephalosporins (cephalexin, cephradine, cefaclor). Pseudomonas spp., Acinetobacter spp., and the enterococci were resistant to U-76,253A and the other tested drugs. Broth microdilution MIC quality control (QC) limits were established for U-76,253A in a multilaboratory investigation using a minimum of 125 MIC determinations per organism. The following MIC QC ranges were recommended; Escherichia coli (ATCC 25922) = 0.25-1.0 micrograms/ml, Staphylococcus aureus (ATCC 29213) = 2.0-4.0 micrograms/ml and Pseudomonas aeruginosa (ATCC 27853) = greater than 32 micrograms/ml.

Cefdinir (FK482), an orally administered cephalosporin in vitro activity comparison against recent clinical isolates from five medical centers and determination of MIC quality control guidelines.

Cefdinir, a new oral cephalosporin, was compared to cefaclor, cefadroxil, cefixime, and cefuroxime against greater than 5000 recent aerobic clinical isolates. This multicenter study revealed broad-spectrum cefdinir activity against all Enterobacteriaceae (MIC50s, 0.06-2 micrograms/ml) except Enterobacter cloacae, Morganella morganii, Proteus vulgaris, and Serratia marcescens (MIC50s, greater than or equal to 4 micrograms/ml). Oxacillin-susceptible staphylococci (MIC90s, 0.5-2 micrograms/ml), beta-hemolytic Streptococcus group B (MIC90, 0.06 micrograms/ml), and Acinetobacter lwoffii were also susceptible to cefdinir. The activity of cefdinir was similar to that of cefixime and cefuroxime against Gram-negative organisms and superior to all tested oral cephems when tested against Gram-positive cocci. None of the cephalosporins were active against oxacillin-resistant Staphylococcus spp., enterococci, Pseudomonas spp., or Xanthomonas maltophilia. MIC quality control range guidelines were established for the strains recommended by the National Committee for Clinical Laboratory Standards documents.

Antimicrobial activity of cefpirome. An update compared to five third-generation cephalosporins against nearly 6000 recent clinical isolates from five medical centers.

Cefpirome, cefotaxime, ceftazidime, cefoperazone, ceftizoxime, and ceftriaxone were tested against approximately 6000 fresh clinical isolates from five medical centers. For 3031 strains of Enterobacteriaceae tested, cefpirome consistently had the lowest MIC50s and lowest percentage of resistant strains. Cefpirome was also the most active agent against the 2138 Gram-positive cocci tested; Staphylococcus haemolyticus was uniformly resistant to all agents tested. Against 791 nonenteric Gram-negative bacilli, the activity of cefpirome was most comparable to that of cefoperazone and slightly less active than ceftazidime. Among the current third-generation cephalosporins, cefotaxime and cefoperazone emerged as having better overall balanced activity. Ceftazidime displayed poorest coverage against Enterobacteriaceae and Gram-positive organisms. Ceftizoxime also provided compromised coverage of staphylococci and nonenteric Gram-negative bacilli. Cefpirome remains as active as originally described in 1984 and possesses a slightly wider spectrum of activity against contemporary aerobic pathogens compared to currently marketed third-generation cephalosporins.

Reproducibility of broth microdilution and disk diffusion susceptibility tests of nine antimicrobial agents against Streptococcus pneumoniae ATCC 49619.

Collaborative studies documented the reproducibility of broth microdilution susceptibility tests of Streptococcus pneumoniae against nine antimicrobial agents and of disk diffusion tests with six of those drugs. Replicate tests of Streptococcus pneumoniae ATCC 49619 in five different laboratories led to the following provisional quality control limits: cefdinir--0.03 to 0.25 microgram/ml and 26 to 31 mm; cefetamet--0.5 to 2 micrograms/ml and 20 to 25 mm; ciprofloxacin--0.25 to 1 microgram/ml and 20 to 26 mm; clinafloxacin--0.03 to 0.125 microgram/ml and 28 to 34 mm; grepafloxacin--0.06 to 0.5 microgram/ml and 21 to 28 mm; PD131628--0.125 to 0.5 microgram/ml and 24 to 29 mm; clindamycin--0.03 to 0.12 microgram/ml; cefpodoxime--0.03 to 0.12 microgram/ml; and trospectomycin--1 to 4 microgram/ml (disk tests were not evaluate for the latter three drugs).

Comparative antimicrobial activity of piperacillin-tazobactam tested against more than 5000 recent clinical isolates from five medical centers. A reevaluation after five years.

Piperacillin combined with tazobactam at a fixed concentration (4 micrograms/ml) and a ratio (8:1) was tested against 5,029 aerobic isolates and 447 fastidious organisms, including anaerobes. Among the Enterobacteriaceae, > 95% inhibition was shared only by imipenem (99.1% at < or = 4 micrograms/ml), and some newer cephalosporins (95.1% - 99.8% at < or = 8 micrograms/ml), and piperacillin-tazobactam (95.8% at < or = 16/4 micrograms/ml). Piperacillin-tazobactam was the most active agent tested against nonenteric Gram-negative bacilli (93.5% at < or = 8 micrograms/ml). Ampicillin-sulbactam was the most active agent against staphylococci (95.0% at < or = 8 micrograms/ml), followed by imipenem (91.8%), piperacillin-tazobactam (89.3% at < or = 8/4 micrograms/ml), and cefepime (86.2% at < or = 8 micrograms/ml). Against the enterococci, only ampicillin (93.0% at < or = 8 micrograms/ml) with or without sulbactam, piperacillin (91.0% at < or = 16 micrograms/ml) with or without tazobactam, and imipenem (91.0%) had acceptable activity. Piperacillin-tazobactam and imipenem were the most active drugs tested against all aerobic isolates, inhibiting 93.5% of isolates each. Piperacillin-tazobactam inhibited all fastidious isolates tested, including Haemophilus influenzae (MIC90, 0.094/4 micrograms/ml), Moraxella catarrhalis (MIC90, 0.064/4 micrograms/ml), Neisseira gonorrhoeae (MIC90, < or = 0.016/4 micrograms/ml), and Streptococcus pneumoniae (all MICs, < or = 4/4 micrograms/ml). Against the anaerobic isolates, the most broad-spectrum antimicrobial agents tested were imipenem (100.0%), piperacillin-tazobactam (99.5% at < or = 32/4 micrograms/ml), metronidazole (98.4% at < or = 8 micrograms/ml), and ticarcillin-clavulanic acid (95.1% at < or = 32/2 micrograms/ml). These results are nearly identical to a previous study involving the same five medical centers in 1989. Piperacillin-tazobactam appears to remain a highly effective beta-lactamase inhibitor combination with a wide empiric spectrum and potency in teaching hospitals.

Tentative criteria for determining the in vitro susceptibilities of Haemophilus influenzae, including quality control parameters, to two fluoroquinolones (grepafloxacin and PD 131628).

Grepafloxacin (OPC 17116) and PD 131628 were evaluated against 150 Haemophilus influenzae isolates to propose susceptibility testing criteria for both broth dilution and disk diffusion procedures using haemophilus test medium. Grepafloxacin-susceptible isolates are defined as those for which minimum inhibitory concentrations (MICs) are < or = 0.06 micrograms/ml and zones of inhibition are > or = 27 mm. PD 131628-susceptible strains of H. influenzae included those that exhibited MICs < or = 0.03 micrograms/ml, and the zones were > or = 32 mm. All MICs were well below concentrations that can be achieved in the blood and tissues of patients treated with either study drug. Criteria for defining a resistant category cannot be determined until strains with elevated MICs are available for study. The proposed quality control guidelines for H. influenzae ATCC 49247 and the disk test are 32-39 mm in diameter for grepafloxacin and 34-42 mm for PD 131628. The preliminary broth microdilution MIC control limits for this strain are 0.002-0.016 micrograms/ml for grepafloxacin and 0.002-0.008 micrograms/ml for PD 131628.

Multilaboratory evaluation of the in vitro activity of 13 beta-lactam antibiotics against 1474 clinical isolates of aerobic and anaerobic bacteria.

The in vitro activity of 13 beta-lactam antibiotics against 1474 recent clinical isolates was evaluated in a multilaboratory study. The most active antibiotic tested in this study was imipenem (98.5% of the strains were susceptible), followed by ticarcillin-clavulanate (91.4%), cefoperazone (90.0%), ceftazidime (87.9%), cefotaxime (87.7%), ceftriaxone (87.0%), ceftizoxime (86.3%), cefotetan (78.5%), ampicillin-sulbactam (77.9%), cefoxitin (73.5%), cefuroxime (70.9%), cefonicid (64.5%), and cefazolin (57.9%).