R A Fromtling's research while affiliated with Richmond VA Medical Center and other places
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Publications (79)
Thirteen laboratories collaborated to optimize interlaboratory agreement of results of a broth macrodilution procedure for testing three classes of antifungal drugs against pathogenic yeasts. The activities of amphotericin B, flucytosine, and ketoconazole were tested against 100 coded isolates of Candida albicans, Candida tropicalis, Candida paraps...
A collaborative comparison of macro- and microdilution antifungal susceptibility tests was performed in five laboratories. MICs of amphotericin B, fluconazole, flucytosine, and ketoconazole were determined in all five centers against 95 coded isolates of Candida spp., Cryptococcus neoformans, and Torulopsis glabrata. A standard protocol with the fo...
In screening for antifungal inhibitors from fungi, four related antifungal agents have been isolated from the cultivation of Aspergillus fumigatus ATCC 20857. These agents were initially produced by the microorganism growing on a solid millet-based medium. A liquid medium containing both glucose and glycerol has also been developed in which these a...
Restricticin (1) is a naturally-occurring antifungal agent which contains triene, pyran and glycine ester functionalities and is unrelated to any previously known family of natural products. This unstable compound, as well as its corresponding N,N-dimethyl derivative (2), have been produced and isolated from both solid and liquid fermentations of P...
As a new prodrug approach to norfloxacin (NFLX) we prepared the acetoxyalkyl carbamates of the type NFLX-CO-OCHR-OAc by the reaction of sodium or mercuric acetate on NFLX alpha-chloroalkyl carbamates. These produrgs did not have the bitter taste of NFLX. In vitro, the acetoxyethyl carbamate exhibited activity only against Staphylococcus spp. and wa...
A new beta-1,3-D-glucan synthesis inhibitor, L-687,781 is produced by the cultivation of Dictyochaeta simplex ATCC 20960. L-687,781 exhibits potent in vitro antifungal activity as well as anti-Pneumocystis activity in a rat model.
A multicenter study was performed to evaluate the effect of medium, incubation time (24 and 48 h), and temperature (30 and 35 degrees C) on intra- and interlaboratory variations in MICs of flucytosine, amphotericin B, and ketoconazole for yeasts. Testing was performed on coded isolates of Candida species (11 strains) and Cryptococcus neoformans (2...
Pneumocystis carinii pneumonia is a major cause of death in AIDS patients in the United States. The presently available treatments have limited use due to a high incidence of adverse reactions. Therefore, there is an urgent need for a safer method for treatment and prevention of this disease. Recent evidence has suggested that P. carinii is related...
Efrotomycin is an N-methylhydroxypyridone glycoside antibiotic with activity primarily against Gram-positive bacteria. It is intended for use as a feed additive for swine. Although efrotomycin is unrelated to any antibacterial drug used in human or veterinary medicine, the possibility of cross-resistance with other antibacterials is of concern. The...
Produit par une souche de Fusarium, cet antifongique ne semble pas adapte au traitement de mycoses chez l'homme ou l'animal. Par contre, il possede une forte activite contre des champignons phytopathogenes ou contaminants de produits stockes
Studies on the distribution, epidemiology and pathogenesis of Cryptococcus neoformans on the island of Puerto Rico are few. We have studied mouse virulence and in vitro antifungal susceptibility of 133 isolates of C. neoformans: 121 environmental and 12 clinical (9 from AIDS patients), that were isolated in Puerto Rico. In experimental CD-1 mice in...
Until the present decade, no studies had been conducted in Puerto Rico on the saprophytic distribution and variety of Cryptococcus neoformans. Samples (522) of pigeon droppings from 14 western towns were tested for the presence of C. neoformans. The yeast was recovered from 24.7% (129 isolates) of the samples, representing 10 of the 14 towns studie...
The in vitro antibacterial spectrum of L-658,310, a new semisynthetic cephalosporin, was compared with ceftazidime, aztreonam and piperacillin against a wide variety of randomly selected human clinical isolates. The compound was found to be a broad spectrum bactericidal agent that was more potent than any of the comparison drugs against glucose non...
The therapeutic activity of L-658,310 was demonstrated in experimental bacteremias in normal, diabetic and neutropenic mice. Especially potent activity was shown against the usually difficult to control pathogens, Enterobacter cloacae and Pseudomonas aeruginosa, that were resistant to ceftazidime and/or gentamicin. Pharmacokinetic studies in mice s...
L-671,329 is a novel, echinocandin-like natural product that possesses potent anti-Candida activity, including activity against Candida parapsilosis. The in vitro MICs of L-671,329 were comparable to aculeacin against 18 yeasts and three filamentous fungi in an agar dilution assay. L-671,329 lysed mouse red blood cells (RBCs) at a concentration of...
L-657,398 is a broad spectrum antifungal agent isolated from solid fermentation or from the mycelium of the liquid fermentation of Aspergillus ochraceus. Structurally, the compound is a novel pyrollidine related to anisomycin.
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L-656,575 is a new oxacephem that, based on studies in rhesus monkeys, is expected to have a moderately long half-life in humans. After administration of a 10-mg/kg dose by the intramuscular route to rhesus monkeys, peak serum concentrations of 32-54 micrograms/ml were seen at about 30 minutes, and the half-life was estimated to be 63 minutes. Urin...
L-656,575 (OCP-9-176) is a novel oxacephem superior to ceftazidime in in vitro activity against clinical isolates of Enterobacter species, methicillin-susceptible Staphylococcus aureus and Staphylococcus epidermidis, and multiply-resistant Pseudomonas aeruginosa. Our results suggest that L-656,575 has a high affinity for penicillin binding proteins...
Pathogenicity and virulence factors were studied for a wild-type strain of Candida albicans (MY 1044) and an auxotrophic, temperature-sensitive mutant strain (MY 1049) that was derived by ultraviolet irradiation. The mutant was a temperature-sensitive, serine auxotroph. Renal pathology and chemiluminescence of spleen cells from infected mice were a...
A strain of Cryptococcus neoformans that was isolated from the cerebrospinal fluid of a human diagnosed as having acquired immunodeficiency syndrome (AIDS), and that produced cutaneous lesions in experimentally infected, normal mice is described. Although no unusual cutaneous manifestations were noted in the patient's records, this isolate of C. ne...
Fungal infections are a major burden to the health and welfare of modern humans. They range from simply cosmetic, non-life-threatening skin infections to severe, systemic infections that may lead to significant debilitation or death. The selection of chemotherapeutic agents useful for the treatment of fungal infections is small. In this overview, a...
Macrophages play a key role in two important host defense mechanisms: (1) ingestion and inactivation of invading organisms, and (2) macrophage interactions with other immune cells, i.e., T lymphocytes and B lymphocytes, that promote antigen recognition and processing. Among the many functions of macrophages in host defense, attachment to and engulf...
Opportunistic infections caused by Candida tropicalis have been noted with increasing frequency in compromised patients. The pathogenicity of three isolates of C. tropicalis was studied in normal CD-1 mice, streptozotocin-induced diabetic mice, and cyclophosphamide-induced neutropenic mice. Lethal dose 50% endpoints and quantitative distribution of...
Lysosomotropic detergents, which kill mammalian cells by disrupting lysosomal membranes, have now been found to be antifungals also. All strains in our assay are susceptible. The mode of action is as yet undetermined, but intracellular vacuoles may be the primary targets.
The luminol-enhanced chemiluminescence (CL) assay was used to examine the effects of antifungal agents tested at concentrations
above and below therapeutically achievable levels on the CL response of mouse spleen cells. Reduction in the CL response of
phagocytic cells may be indicative of an inhibition of the cellular immune response. Concomitantly...
A model of ascending urinary tract infection due to an isolate of Escherichia coli was developed in normal and streptozotocin-induced diabetic mice to compare the efficacy of norfloxacin and trimethoprim-sulphamethoxazole.
Norfloxacin and trimethoprim-sulphamethoxazole both were effective in reducing the number of colony forming units of E. coli fr...
The focus of this study was to elucidate the influence of lymphocytes on phagocytic cell-dependent chemiluminescence (CL). CL was induced by in vitro addition of mitogens, particulate substances and Keyhole Limpet Hemocyanin (KLH)-antigen. CL was measured in spleen cells of immunologically naive mice and in mice immunized with KLH. Mice immunized w...
The luminol-enhanced chemiluminescence (CL) assay is a measure of the early events of phagocytosis, leukocyte activation, and immune cell interactions. Reduction in the CL response of immune cells may be indicative of an inhibition of the immune response. This study was undertaken to examine the effects of antifungal agents tested at concentrations...
Thirty-nine isolates of Cryptococcus neoformans, nineteen serotype AD and twenty serotype BC, were assayed for susceptibility to eight antifungal agents using an in vitro agar dilution assay. Media employed were Kimmig agar and yeast nitrogen base supplemented with 10% glucose. The antifungal agents used were ketoconazole, amphotericin B, 5-fluoroc...
Chemiluminescence is the result of the respiratory burst generated by phagocytic cells after stimulation by antigen. The measurement of chemiluminescence represents a sensitive means for detecting antigenic stimulation and immune cell function. Although the kinetics of chemiluminescence reactions have been described, appropriate statistical methods...
A review of the literature (148 references) on the interactions of fungi with polymorphonuclear cells, monocytes and macrophages is presented. The interactions of Aspergillus species, Coccidioides immitis, Blastomyces dermatitidis, Histoplasma capsulatum, Cryptococcus neoformans, Candida albicans, and Candida species with human and experimental ani...
Chemiluminescence is a measure of the respiratory burst of phagocytic immune cells following stimulation by antigen. Measurement of chemiluminescence by scintillation counters set in the out of coincidence mode or by instruments designed specifically to measure chemiluminescence is an accurate, reproducible means of evaluating the immune potential...
The in vitro effect of pH and glucose concentration on the antibacterial activity of norfloxacin in urine was studied. Norfloxacin effectively inhibited the growth of four gram-negative pathogens in urine in vitro at pH values of 6.0, 7.0, and 8.0. The antibacterial activity of norfloxacin in urine was reduced severalfold at pH 6, but minimum inhib...
Systemic bacterial infections due to Escherichia coli MB 2884, Proteus mirabilis MB 3125 and Klebsiella pneumoniae MB 4005 were well controlled by treatment with norfloxacin both in normal and streptozotocin-induced diabetic mice. similar observations were made when trimethoprim-sulfamethoxazole was used against susceptible pathogens. Systemic infe...
Bay n 7133 and Bay L 9139 (Bayer AG, Wuppertal, Fed. Rep. Germany) are new, orally absorbable, antifungal imidazole derivatives. In vitro, Bay n 7133 was comparable to ketoconazole and miconazole when tested against isolates of Candida albicans and Cryptococcus neoformans. Bay L 9139 also was active against these organisms but to a lesser degree th...
To evaluate the usefulness of host resistance assays for measurement of immunotoxicologic effects of chemicals, the immunosuppressive effects of exposure to diethylstilbestrol (DES) were compared with the effects of treatment with the known immunosuppressive drug cyclophosphamide (CPS). A panel of six host resistance models was evaluated, including...
Synthetic polyanions have been shown to alter host resistance to infection. The anticryptococcal effect of pyran copolymer was assessed in vivo and in vitro. Pretreatment with pyran copolymer significantly extended mean survival in mice lethally infected with Cryptococcus neoformans when compared to untreated animals (p less than 0.01). The anticry...
Treatment of mice with a single dose of cyclophosphamide 24 h before challenge with Cryptococcus neoformans increased host survival, whereas treatment with 10 daily exposures of cyclophosphamide, starting 2 days before challenge,
markedly reduced survival in mice challenged on the second day of drug treatment. Treatment with 14 daily exposures of d...
An induced mutant of Candida albicans with greatly decreased virulence for mice is described. The mutant was one of five auxotrophic mutants obtained by ultraviolet irradiation of a clinical isolate (strain MY 1044). The five mutants included two methionine auxotrophs, one methionine-cysteine auxotroph, one temperature-sensitive serine auxotroph, a...
Disulfiram (Antabuse)-like reactions occurring in patients undergoing antibacterial therapy with certain cephalosporin antibiotics, and after ingestion of ethanol, are well documented. A murine model is described which may prove useful in the detection of compounds likely to produce this effect. Eight cephem compounds (moxalactam, cefamandole, cefm...
The inhibitory activities of 2 new orally absorbed antifungal imidazole derivatives, BAY n 7133 and BAY 1 9139, were compared in vitro with those of ketoconazole and miconazole. Clinical isolates of pathogenic fungi tested included 35 yeasts, 62 dimorphic fungal pathogens, 37 filamentous fungi and 31 dermatophytes. LY 121019, a semisynthetic analog...
The antibacterial activity of norfloxacin, an organic acid structurally related to nalidixic acid, was compared with that of the oral cephalosporins cefaclor and cephalexin, and with that of nalidixic acid, cinoxacin, amikacin, ampicillin, trimethoprim alone and the combination of trimethoprim and sulfamethoxazole. Agar dilution studies were perfor...
Immune responses, including lymphocyte transformation and protection, were evaluated in a new model of experimental murine cryptococcosis. Newly developed, genetically stable, acapsular mutants of Cryptococcus neoformans were used to immunize C3H/HeN mice. Intraperitoneal and subcutaneous routes of immunization without adjuvant were studied. Spleen...
The inhibitory activities of 2 new orally absorbed antifungal imidazole derivatives, BAY n 7133 and BAY l 9139, were compared in vitro with those of ketoconazole and miconazole. Clinical isolates of pathogenic fungi tested included 35 yeasts, 62 dimorphic fungal pathogens, 37 filamentous fungi and 31 dermatophytes. LY 121019, a semisynthetic analog...
From the rhizosphere of a potted Sanseviera trifasciata, a Streptomyces griseus strain with a broad antifungal activity was isolated; in the soil of this plant, a sole hyphomycete, Aspergillus niger, was found in pure culture at 37 degrees C. This streptomycete produced a polyene, belonging to the pentaene group. It was found to inhibit, in vitro,...
Of 90 clinical isolates of Cryptococcus neoformans studied, 3 were determined to be serotype B/C. The patients from whom these B/C isolates were obtained were identified as never having lived in or visited the areas associated with B/C serotypes. This finding suggests a broader geographic distribution of this serotype group than previously believed...
Six acapsular strains of Cryptococcus neoformans obtained by chemical mutagenesis failed to produce a capsule in vivo and were avirulent in mice following high dose intramuscular, intraperitoneal or intravenous inoculation. Peritoneal granulomas were observed in all animals inoculated with the acapsular mutants. These granulomas were characterized...
Clearance of cryptococcal polysaccharide (CP) from tissues and body fluids of nonimmune mice was studied. Mice were injected intravenously only with one mg of purified CP, and serum, urine and tissues were obtained from each animal at various intervals for a period of 84 days. Tissue extracts, serum and urine were tested for CP content by enzyme-li...
Cryptococcosis is an often fatal opportunistic fungal infection. Despite efforts to elucidate the role of immunity in host defense against the disease, much remains to be learned. The purpose of this brief review is to provide the reader with an overview of the history of research concerned with host immunity in cryptococcosis. both humoral and cel...
An examination of 21 strains of Cryptococcus neoformans isolated from environmental and clinical sources in Germany revealed only serotypes A, D, and AD. Of these, 13 isolates were serotype A, 5 isolates were serotype D, and 3 isolates were identified as serotype AD. The absence of serotypes B or C confirms earlier reports from other European count...
The effect of uremia on immune incompetence was studied. BALB/c mice were infected with a minimally virulent strain of Cryptococcus neoformans 6 weeks before immune assay. Uremia was induced by intramuscular injection of 0.15 ml of glycerol. Pooled spleen cells from four experimental groups (normal, uremic, infected, and infected and uremic) were a...
Pigeon droppings in a vacant tower were assayed for the number and size of viable cells of Cryptococcus neoformans. The dry, thinly scattered floor debris contained 2.6 x 10(6) viable cells per g--300 times more cells than were cultured from a large, compact pile of pigeon droppings (7.4 x 10(3) cells per g). Aerosols generated from floor debris co...
Three pseudohyphal isolates of Cryptococcus neoformans were inoculated intracranially into mice. Four weeks post-inoculation the animals showed no symptoms of disease and the number of viable cells per brain decreased to zero. Possible roles of pseudohyphal forms of C. neoformans in the immunology and pathogenesis of cryptococcosis are discussed.
A modified blood residual nitrogen plate auxanographic method was applied to the detection of experimental uremia in a murine model. The yeast-like fungus, Cryptococcus neoformans, was used as the indicator. Transient uremia was induced by injection of 0.2 ml glycerol intramuscularly. The low molecular weight nitrogen levels were estimated by measu...
Eighty clinical and 28 soil isolates of C. neoformans obtained in Oklahoma were separated into A-D and B-C serotype groups utilizing creatinine-dextrose agar with bromthymol blue. Previously, serotype B-C clinical isolates have been frequent only in patients from Southern California where as many as 50% of the isolates are of this type. In contrast...
The entomopathogenic fungus, Beauveria bassiana, was isolated from pulmonary lesions of a dead American alligator (Alligator mississipiensis) at the Oklahoma City Zoo. Colonies of the fungus, which had sporulated in vivo, were found in the thoracic air spaces. Septate, branching hyphae and fungal spores were seen in stained histologic sections of p...
Mice were immunized with a viable, avirulent strain ofCryptococcus neoformans. Lymphocyte blastogenic assays showed a 10-fold increase in reactivity of sensitized spleen cells, and histopathologic examination revealed marked splenic hyperplasia. Thirty-two days after intravenous inoculation with a virulent strain ofC. neoformans, none of the contro...
Fatal pulmonary disease occurred in two captive American alligators. The entomopathogenic fungus, Beauveria bassiana, was isolated from pulmonary lesions in both alligators. An extended hibernation period because of a severe winter and a failure of the zoo heating system may have predisposed the alligators to infection.
A murine model was developed to study the cell-mediated immune response of mice immunized with one of two live, avirulent forms of Cryptococcus neoformans: a nonencapsulated mutant and a thinly encapsulated pseudohyphal variant. A lymphocyte transformation assay was used to evaluate the cellular response of control and sensitized spleen cells after...
The antifungal effect of an aqueous extract of garlic was tested against 18 strains of Cryptococcus neoformans. Nine strains were clinical isolates, eight were laboratory-induced pseudohyphal variants, and one was an ultraviolet-radiation-induced, nonencapsulated mutant. Saturated-disk sensitivity tests showed average zones of inhibition of 37.9 mm...
The antifungal effect of an aqueous extract of garlic was tested against 18 strains of Cryptococcus neoformans. Nine strains were clinical isolates, eight were laboratory-induced pseudohyphal variants, and one was an ultraviolet-radiation-induced, nonencapsulated mutant. Saturated-disk sensitivity tests showed average zones of inhibition of 37.9 mm...
Although cryptococcosis is characterized as a chronic central nervous system disease, it is generally accepted that the lungs are the primary portal of entry for the etiological agent. Despite this, there is a distinct lack of evidence that viable airborne particles of Cryptococcus neoformans are small enough to reach the alveoli. Two encapsulated...
Clinical evidence suggests that macular disease may occasionally be a result of infection by Histoplasma capsulatum. Experiments were performed to study the induction of ocular lesions by this fungus. A group of Dutch Belted rabbits was immunized with killed mycelial antigen, rested, and then challenged with live yeast antigen. Another group of rab...
responseof spleen cells was investigated after stimulation withtheparticulate antigen zymosan, recommendations are madeforthedesign andstatistical evaluation ofsuchstudies. Threeparameters were usedinassessing the chemiluminescence response;peakintensity oftheemitted light, timetopeak,andthearea underthe intensity-time curve.Thedataindicated thatpe...
Citations
... The in vitro and in vivo inhibition of the topoisomerase II enzyme of the DNA, alike the quinolones, suggest an interference in the RNA and protein synthesis [38]. Furthermore, melanin, produced by the action of phenol oxidase on L-DOPA [39,40], is a virulence factor for fungi of the Cryptococcus genus for protecting them from oxidative stress, phagocytosis, the action of antifungal compounds, as well as for modifying the host immune response [41][42][43][44][45][46]. As liriodenine inhibits the melanin synthesis in PC-12 cells, this antimelanin activity could play a role in its anticryptococcal activity [47]. ...
... The Clinical Laboratory Standards Institute (CLSI) for yeasts (M27-A3) method was used to estimate the minimum inhibitory concentration (MIC) of the test compound and the drug [30]. The MIC was determined in an RPMI-164 medium (Cat no. ...
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... Morphogenesis profoundly shapes Cryptococcus interaction with various hosts (12,19). Although the inverse relationship between Cryptococcus filamentation and its virulence in mammalian hosts have been observed since the 1960s and 1970s (34)(35)(36)(37)(38)(39)(40)(41)(42), the molecular bases for attenuated virulence of the filamentous form are unknown. We have previously discovered that the transcription factor Znf2 controls cryptococcal yeast-to-hypha transition and is a potent anti-virulence factor (20,21). ...
... typically causes symptoms from the pulmonary and central nervous systems in human immunodeficiency virus (HIV) patients. The typical scenario starts with the patient inhaling Cryptococcus, which is then transmitted through the blood circulation to various organs and sites in the body, with a particular preference for the central nervous system [3]. Cryptococcus neoformans is the most prevalent form worldwide. ...
... Garlic suppresses beneficial intestinal microflora as well. However, probably due to a greater sensitivity of enterobacteria to allicin , it is more effective against potentially harmful entero- bacteria [106][107][108][109][110][111][112][113][114][115]. Garlic extract with high content of ajoene is significantly effective against Candida albicans, Paracoccidiodes, Aspergillus sp., Botrytis cinerea and Trichoderma harzianum Scedosporium prolificans, Tinea pedis, Opalina ranarum, Entamoeba histolytica, Leishmania, Trypanosomes, Leptomonas, Crithidia, etc. [38, 116,117] . ...
... Morphogenesis profoundly shapes Cryptococcus interaction with various hosts (12,19). Although the inverse relationship between Cryptococcus filamentation and its virulence in mammalian hosts have been observed since the 1960s and 1970s (34)(35)(36)(37)(38)(39)(40)(41)(42), the molecular bases for attenuated virulence of the filamentous form are unknown. We have previously discovered that the transcription factor Znf2 controls cryptococcal yeast-to-hypha transition and is a potent anti-virulence factor (20,21). ...
... . Species such as Fusarium solani, Fusarium moniliforme, Fusarium verticillioides, Aspergillus ustus, Aspergillus fumigatus, Beauveria bassiana, Acremonium (formerly Cephalosporium), Candida albicans, Metarhizium spp., Mucor spp., Paecilomyces farinosus, Paecilomyces lilacinus, Penicillium spp., Stachybotrys spp., Chaetomium spp., Mucor spp., Rhizopus spp. have been linked to infections in crocodiles[1,6,11,[13][14][15][16][17][18][19][20]. ...
... Examples of toxins produced include destruxins and beauvericin; in general, studies have shown low levels of toxicity in vertebrates, with some exceptions (for example, mice are susceptible to the toxic effects of destruxin A). 28,29 These fungi are largely considered nonpathogenic to vertebrates, although there are isolated reports of infection in both humans and animals. 3,[5][6][7][8][9]14,15,17,[21][22][23][24]28,29 This report describes eight cases of fatal pulmonary mycosis and one case of fatal mycotic coelomitis in eight different reptile species from two orders in North America due to Metarhizium or Beauveria infection. ...
... Seven different antifungal agents (Fluconazole, Amphotericin B, Flucytosine, Itraconazole Micafungin, Posaconazole, and Voriconazole) were selected for C. albicans antifungal susceptibility testing using the broth microdilution test according to the recommendation of the Clinical Laboratory Standards Institute (29). The MIC breakpoints were interpreted (30,31). ...
... 2 Sphingofungins are a family of promising bioactive molecules first discovered in 1992, acting as sphingolipid inhibitors (SIs). 3 They have gained interest for their ability to inhibit serine palmitoyl transferase (SPT) which catalyzes the primary reaction of sphingolipid biosynthesis, that is, the condensation of serine with palmitoyl-CoA. 4 Sphingofungins A-D were first isolated from the human pathogenic fungus Aspergillus fumigatus. 3 Shortly thereafter, sphingofungins E and F were isolated from Paecilomyces variotii 5 and recently, sphingofungins G and H have been reported in Aspergillus penicillioides 6 ( Figure 1A). ...
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