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L Guelfand

Publications and source records attributed to L Guelfand.

10 recordsLinked to original sources

Timed-kill curves for Cryptococcus neoformans isolated from patients with AIDS.

Infection with Cryptococcus neoformans is an increasing problem in immunocompromised patients, particularly those with acquired immune deficiency syndrome (AIDS). Amphotericin B and fluconazole are currently acceptable therapies for cryptococcal meningitis; however, their effects remain suboptimal and recurrence or treatment failure is still a problem. Antifungal susceptibility testing may be an important tool for guiding therapy, but for C. neoformans, a reliable method is still not available. This retrospective study evaluated minimal inhibitory concentration (MIC) for amphotericin B and fluconazole, and minimal fungicidal concentration (MFC) and timed-kill curves for amphotericin B against 16 clinical isolates of C. neoformans obtained from AIDS patients with cryptococcal meningitis. No correlation between clinical outcome and MIC was observed for amphotericin B. In selected cases, the MFC seemed to be a better predictor of outcome than MIC. In this study, amphotericin B timed-kill curves appeared to show a correlation with clinical outcome of the 16 patients with AIDS-associated cryptococcal meningitis. These in vitro tests must be further evaluated in prospective studies to confirm their potential usefulness for guiding cryptococcal meningitis therapy.

AIDS-Related Opportunistic Infections↗

In vitro susceptibility studies of Cryptococcus neoformans isolated from patients with no clinical response to amphotericin B therapy.

The in vitro activities of three antifungal drugs alone and in combination were evaluated against five isolates of Cryptococcus neoformans using time-kill curves (TKC). The isolates were from AIDS patients who had either died or had failed to show a clinical response during amphotericin B (AMB) treatment. AMB, fluconazole (FCZ) and flucytosine (5FC), and combinations of the drugs (AMB plus 5FC, AMB plus rifampicin (RIF) and FCZ plus 5FC), were evaluated. With all five isolates AMB did not show fungicidal activity; instead, a persistent or tolerant effect was observed. Combinations of AMB plus 5FC and AMB plus RIF showed a clear synergic effect, except for one isolate tested with AMB plus RIF. In contrast, the FCZ plus 5FC combination did not inhibit growth of any isolate.

Acquired Immunodeficiency Syndrome↗

[In vitro activity of trovafloxacin, of other fluoroquinolones and of related antimicrobials against clinical isolates. Grupo colaborativo WHONET-Argentina].

The in vitro activity of trovafloxacin (TRV) has been evaluated in comparison with that of other antimicrobial agents against 5671 clinical isolates recovered by representative institutions of different provinces in our country. The resistance percentage to gentamicin and third generation cephalosporins among enterobacteriaceae was high: 17% and 16% respectively, with a considerable variation according to the analyzed species. The resistance to ciprofloxacin (CIP) and TRV affected approximately 9% of the isolates, without significant differences between both drugs. Fluoroquinolones (FQ) presented excellent activity on 166 isolates of Salmonella spp., 208 of Shigella flexneri and 76 of Shigella sonnei, where only one S.sonnei isolate was resistant to CIP, but susceptible to TRV. About half the isolates of Salmonella spp. and S.sonnei and almost all S.flexneri isolates were resistant to ampicillin, and more than 60% of Shigella spp. isolates were resistant to trimethoprim-sulfamethoxazole. A 41% of Staphylococcus aureus and 55% of coagulase-negative staphylococci isolates were resistant to oxacillin, presenting a highly associated multi-resistance. The resistance to FQ was also strongly related to oxacillin resistance, but the resistance to TRV was significantly lower than the CIP resistance: 9% vs 57% for S.aureus and 4% vs 41% for coagulase-negative staphylococci. A similar behavior was observed with Enterococcus spp., where 54% of the isolates were resistant to norfloxacin and only 13% were resistant to TRV. Neither Streptococcus pneumoniae (n = 193) nor Haemophilus influenzae (n = 139) isolates presented resistant to TRV.

Anti-Bacterial Agents↗

[Diarrhea and AIDS: more complex diagnostic techniques; better therapeutic results].

During 17 months, 73 HIV-positive patients with diarrhea lasting at least for 14 days, were studied prospectively. The patients had stool specimen examinations negative for enteric pathogens, or positive for one of them, but with no response to specific treatment. All patients were subcomitted to digestive endoscopy and biopsies were taken for microbiological and histological studies. The etiology of the diarrhea could be established in 48 patients (66%). In 45, the cause was an enteric infection. There was association of 2 pathogens in 11 patients, and of 3 in 1 patient. The agents found were: Cryptosporidium (24%), MAI (16%), Giardia lambila (12%), isospora belli (5%), Shigella (5%), Salmonella (5%); Entamoeba histolytica (3%), HSV (3%), tuberculosis (2%), adherent bacteria (2%) and spirochetes (2%). In 3 patients the etiology was not infection, their diagnoses were coeliac disease, lymphoma and idiophatic colonic ulcers, respectively. In 51% of the cases only the examination of endoscopic biopsy specimens could identify the cause of the diarrhea. These results justify the use of these methods to improve diagnosis and therapeutic attempts in these patients.

AIDS-Related Opportunistic Infections↗

[Utility of prolonged incubation and terminal subcultures of blood cultures from immunocompromised patients].

The value of blind terminal subcultures (7 and 30 days) and prolonged incubation (30 days) of blood cultures from immunosuppressed patients was analyzed in the Fundación Favaloro, the Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia and the Hospital de Niños Ricardo Gutiérrez. A total of 2707 blood cultures and 369 patients were included (transplantation of solid organs 154, oncohematologic disorders 106 and solid tumors 109). Bact-Alert bottles were incubated at 35 degrees C for 30 days in the Bact-Alert System. Bottles with positive signals were routinely removed, and aliquots of the broth were Gram stained and subcultured aerobically in chocolate agar and Sabouraud agar. A total of 136 bacteremic episodes were obtained. The positivization time of blood cultures was 81.6% at 24 h, 93.3% at 48 h, 94.5% at 72 h and 97.7% within 7 days. Only 3 (2.2%) episodes were positive by blind terminal subcultures and 1 (0.75%) by prolonged incubation (14 days). The median time and range of positivization in hours were 13.8 and 2.2-168, respectively. The microorganisms isolated were coagulase negative staphylococci (n = 24), Klebsiella pneumoniae (n = 22), Staphylococcus aureus (n = 21), Escherichia coli (n = 18), Acinetobacter spp (n = 9), Candida spp (n = 8), Pseudomonas aeruginosa (n = 6), Enterobacter cloacae (n = 5), Stenotrophomonas maltophilia (n = 5), Enterococcus faecalis, Salmonella spp and Capnocytophaga sputigena (n = 2), Enterobacter aerogenes, Enterococcus faecium, Citrobacter diversus, Candida albicans, Klebsiella oxytoca, Chryseomonas luteola, Serratia marcescens, Abiotrophia spp, Campylobacter jejuni, Moraxella catarrhalis, Moraxella urethralis, Neisseria sicca, beta hemolytic group G streptococci, Rhodococcus equi, Micrococcus spp, Cryptococcus neoformans and Streptococcus mitis (n = 1). In our experience, blind terminal subcultures and prolonged incubation of blood cultures from immunosuppressed patients are unnecessary and cost expensive.

Bacteremia↗

[Quantitative cultures from catheters, using the Brun Buisson technique].

The objective of this collaborative work carried out in the Fundación Favaloro and the Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia, was to determine optimal conditions for incubation (time and atmosphere) of quantitative cultures of catheters processed according to the technique of vortex agitation (Brun Buisson method). From 689 processed catheters, 551 yielded negative cultures. From the 138 positive cultures, 125 yielded monomicrobial cultures and 13 polimicrobial cultures (total number of microorganisms was 151). In the last situation each micoorganism was considered on an individual basis. A total of 58 episodes of catheter related bacteremias occurred, being 52 monomicrobial and 6 polimicrobial (total number of microorganisms was 64). When colony counts were compared in aerobic and in 5-10% CO2 atmospheres, a very good correlation was obtained (p = 0.27; r2 = 0.9268). No advantage was observed by incubating plates for more than 48 hours. Colony counts performed at the second versus the third day, and at the second day versus the seventh, gave very good correlation (p = 0.10 and r2 = 0.9996; p = 0.31 and r2 = 0.9995, respectively).

Aerobiosis↗

[Comparison of methods for the identification of the most common yeasts in the clinical microbiology laboratory].

We evaluated different methods for the routine identification of medically important yeasts. A total of 150 clinical isolates: 25 C. albicans, 25 C. tropicalis, 25 C. glabrata, 25 C. parapsilosis, 8 C. guilliermondii, 11 C. krusei and 31 Cryptococcus neoformans were tested by Yeast Biochemical Card bioMerieux Vitek (YBC), CHROMagar Candida (CHR). The addition of yeast morphology in Corn Meal agar-Tween 80 (AM) to YBC and CHR was also evaluated. The reference methods used were: API 20C, germ tube formation, AM, Christensen urea and Birdseed agar. YBC identified 135 yeasts with an overall accuracy of 90%. Sensitivity (S) and specificity (E) were: 92-98% for C. albicans and C. tropicalis; 84-99% for C. papapsilosis; 100-99% for C. glabrata; 91-100% for C. krusei; 63-98% for C. guilliermondii and 90-99% for Cryptococcus neoformans, respectively. CHR identified correctly 100% for C. albicans, 92% for C. tropicalis and 91% for C. krusei. Both methods combined with AM provided 100% S and E. We found that YBC system was appropriate for identification of yeasts isolated from human sources. CHR was effective and easy to use for identification of C. albicans, C. tropicalis and C. krusei. The routine use of AM with both methods is recommended.

Automation↗

[Histoplasma capsulatum with aberrant morphology isolated in Argentina].

For the first time in Argentina, we describe a strain of Histoplasma capsulatum var. capsulatum with an aberrant morphology that was isolated from a single patient with AIDS. Mycelial phase cultures on agar Sabouraud at 25-28 degrees C showed white, glabrous, umbilicated and centrally radiated colonies. Unusual microscopic findings were the absence of typical conidia, the presence of terminal/intercalary chlamydoconidia with a diameter of 4 pm and of thickened hyphae. Fungal identification was confirmed by the detection of bands H and M species specific antigens in mycelial culture supernatants and reversion to the typical yeast phase on agar brain-heart-cysteine at 37 degrees C. The genomic DNA profile obtained by RAPD-PCR with primers 1281-1283 coincided with the predominant profile of H. capsulatum among isolates from Argentine patients.

AIDS-Related Opportunistic Infections↗

[Disk diffusion method for fluconazole susceptibility testing of Candida spp. isolates].

In order to standardize and evaluate a disk diffusion method with visual reading to detect in vitro fluconazole susceptibility of yeast, 1193 clinical isolates were tested. These included 584 Candida albicans, 196 Candida parapsilosis, 200 Candida tropicalis, 113 Candida glabrata, 50 Candida krusei and 50 Candida spp. and other opportunistic yeasts. The disks were manufactured in the INEI-ANLIS "Dr. Carlos G. Malbrán. The disk diffusion method results were compared to MIC results obtained by the reference CLSI M27-A2 broth microdilution method modified by EUCAST. The interpretative breakpoints for in vitro susceptibility testing of fluconazole were established at: zone diameter > or =16 mm for MIC < or =8 microg/ml (susceptible isolates), between 9 and 15 mm for MIC = 16-32 microg/ml (susceptible dose-dependent isolates), and < or =8 mm for MIC > or =64 microg/ml (resistant isolates). Overall agreement between the two methods was 94.7%, with 0.2% very major errors, and 0.3% major errors. Inter - and intralaboratory agreement was good. The disk diffusion method for drug susceptibility testing of Candida spp. isolates is inexpensive, reliable and reproducible. However, when the inhibition zone diameter is < or =15 mm, it is advisable to test the isolate by the reference microdilution method.

Antifungal Agents↗