J Cardiovasc Surg (Torino), 2001 Jun, 42(3), 339 - 43 Predictive value of Candida colonization index in 131 patients undergoing two different cardiovascular surgical procedures; Yazdanparast K et al.; BACKGROUND: Nosocomial candidemia is an important infection because of its increasing incidence and its high fatality rate . Candidiasis involves multiple risk factors . In this work, we study the degree of Candida colonization in cardiovascular surgical patients by taking into account the number of sites colonized and the density of growth . METHODS: Eleven (11) selected variables (age, sex, weight, diabetes, number of antibiotics, duration of antibiotics, length of hospitalization, length of stay in surgical intensive care unit, duration of surgery, temperature during surgery, and number of bypass) were considered to predict the perioperative variation of the colonization index (CI) by Candida in 131 cardiovascular surgical patients . These patients were divided into two groups: group A, coronary artery bypass grafting with extracorporeal circulation (72 patients) and group B, coronary artery bypass grafting without extracorporeal circulation (59 patients) . RESULTS: One thousand and forty-eight fungal cultures were obtained from four different body sites and 162 isolated were identified . Candida albicans accounted for 74% of the strains in group A and 97% in group B . The statistical analysis (two-way anova) shows that group A patients with an increased CI have received significantly more antibiotics than those with a stable CI (1.50+/-0.83 vs 1.08+/- 0.40, p=0.0055) . CONCLUSIONS: Epidemiological data obtained from this study show that coronary artery bypass grafting with extracorporeal circulation procedure is associated with an increase in the use of antibiotics and subsequently a higher risk a Candida colonization-infection. J Bacteriol, 2001 Jul, 183(13), 4090 - 3 Efg1, a morphogenetic regulator in Candida albicans, is a sequence-specific DNA binding protein; Leng P et al.; Efg1 is essential for hyphal development in the human pathogen Candida albicans under most conditions . Efg1 is related to basic helix-loop-helix regulators, and therefore most workers presume that Efg1 is a transcription factor . Here we confirm that Efg1 is a DNA binding protein that can interact specifically with the E box. Microbios, 2001, 105(411), 87 - 101 Candida albicans cell wall lytic enzyme produced by Streptomyces thermodiastaticus; Mansour FA et al.; The production of lytic enzyme by Streptomyces thermodiastaticus was found to be affected by some growth conditions and nutritional factors . The highest enzyme production was obtained after 18 h of incubation at pH 5.5 and at 50 degrees C . The carbon source influenced the lytic enzyme production . A higher enzyme yield was obtained when Candida albicans cell wall (1 g/100 ml) was used as the sole carbon source . NaNO3 at 0.1 g/100 ml was the best nitrogen source for enzyme production . From all phosphorous sources, microelements, and growth factors tested, KH2PO4 (1 g/l), ZnSO4 (1 mg/I) and Tween 80 (0.1%), respectively, were found to favour the highest production of lytic enzymes by S . thermodiastaticus . The lytic enzymes mainly produced chitinolytic and proteolytic activities. Mycopathologia, 2001, 150(1), 9 - 13 Onychomycosis in Tehran, Iran: prevailing fungi and treatment with itraconazole; Khosravi AR et al.; A total of 187 Patients with suspected onychomycosis were examined for causative fungal agents between 1996 and 1997 . Laboratory examination confirmed onychomycosis in 115 patients, of which 97 cases were presented with positive microscopic and cultural examinations, and they were selected for itraconazole pulse therapy . From an etiological point of view, 48.4% of the nail infections, mainly toenail infections, were caused by dermatophytes, 43.3% were infected with Candida spp, specially infected fingernails, and 8.2% by non-dermatophytic molds . Trichophyton mentagrophytes var . interdigital and T . violaceum were the most prevalent species . Candida albicans and C . parapsilosis were the predominant species of the Genus Candida . Scopolariopsis brevicaulis was the most common non-dermatophyte molds observed . Female affected more frequently than male and in both sexes, those who were 30-49 years old, more infected . Toenails were affected more frequently than fingernails . In this study, itraconazole pulse therapy (400 mg daily) gave during the first week of per month for 3 months . The study included 51 patients with toenail onychomychosis (group 1) and 46 patients with fingernail infections (group 2) . Patients were followed up for 9 months after the last treatment . Clinical response rates were 83% in the group 1, 95% in the group 2 at month 12; the corresponding mycological cure rates were 71 and 87%, respectively. Microbiology, 2001 Jun, 147(Pt 6), 1687 - 92 Evidence for a more recently evolved clade within a Candida albicans North American population; Lott TJ et al.; Candida albicans is diploid and displays a primarily clonal mode of reproduction . There is, however, evidence for meiosis and the degree to which this occurs in nature is unknown . Although random mating would act to obscure clonal lineages, previous studies have demonstrated that collections of North American isolates display three major partitions with no evidence of geographic clustering . To better understand the extent of sexuality and its role in the phylogeny of the species, a reference subset of 50 isolates representing this tripartite division was analysed using 1 minisatellite, 5 microsatellites (MSs) and 15 nuclear polymorphisms (NP) . A total of 87 alleles were observed for 21 loci and 12/16 informative loci exhibited a departure from Hardy-Weinberg expectations (G(2)</=0.05) . We did not observe an absolute correlation between MSs and NP, although isolates with identical NP genotypes were correlated with a previously defined, predominant class (putative group I) . The use of additional markers did not give increased support for the tripartite structure of the population . However, (9/19) group I isolates were found to be highly related, differing by only one or a few alleles . Designated subgroup A, the interpretation is that these isolates are related by descent and that they are of a more recent evolutionary origin, diverging from an ancestral group I clone . The reason for their relative abundance in the population is unknown; one possibility is that they may be under positive selection. J Immunol, 2001 Jun 15, 166(12), 7019 - 22 Neutrophil Fc gamma RI as target for immunotherapy of invasive candidiasis; van Spriel AB et al.; Invasive candidiasis represents a life-threatening disease for immunocompromised patients . This study focused on new immunotherapeutic approaches for systemic Candida albicans infections in a human FcgammaRI-transgenic mouse model . FcgammaRI (CD64) is a potent immunoactivating receptor on phagocytic and dendritic cells . In vivo targeting of C . albicans toward neutrophil-FcgammaRI by bispecific Abs and G-CSF effectively protected FcgammaRI-transgenic mice from lethal candidiasis . Nontransgenic mice were not protected, and treatment with bispecific Ab or G-CSF alone did not reduce mortality . Furthermore, infected FcgammaRI-transgenic mice developed high titers of anti-C . albicans IgG, and survival was extended on secondary infection without further treatment . These findings document the capacity of FcgammaRI to initiate potent anti-C . albicans immunity and support the development of FcgammaRI-directed immunotherapy of invasive fungal disease. Clin Chem Lab Med, 2001 Apr, 39(4), 319 - 23 Protein S-100B: a serum marker for ischemic and infectious injury of cerebral tissue; Bertsch T et al.; The S-100B protein is released by injured astrocytes . After passage through a disintegrated blood-brain barrier (BBB) the molecule can be detected in the peripheral circulation . We investigated the association between the extent of brain injury and S-100B concentration in serum in cerebral injury caused by cerebral ischemia and cerebral fungal infection . Study I: The S-100B serum concentration was serially determined in 24 patients with ischemic stroke at 4, 8, 10, 24, 72 hours after the onset of symptoms . We observed that patients with brain lesions larger than 5 cm3 exhibited significantly increased serum levels of S-100B at 10, 24 and 72 hours compared to those with lesion volumes below 5 cm3 . Furthermore, an association between S-100B serum concentration and neurological outcome was observed . Study II: In a mouse model of systemic fungal infection with Candida albicans we observed that serum levels of S-100B increased at day 1 after intravenous infection . At this time we could histologically demonstrate brain tissue injury by invading hyphae which had crossed the BBB . Furthermore, reactive astrogliosis was demonstrated by immunohistochemistry . On day 7 we found a significant decrease of S-100B serum level compared to day 1 and 4 . This was associated with a demarcation of the fungi with leukocytes in brain tissue at this late phase of infection . No further invasion through the BBB was seen on day 7 . In conclusion, serum levels of S-100B reflect the time course of tissue injury in cerebral ischemia and cerebral infection to a similar extent . Thus, S-100B may be a useful marker to assess cerebral tissue injury. Med Microbiol Immunol (Berl), 2001 Apr, 189(3), 169 - 73 Cecropins, antibacterial peptides from insects and mammals, are potently fungicidal against Candida albicans; Andra J et al.; Natural products are the major source of lead compounds for drugs against human pathogens . Among the first natural peptides from animals for which a potent antibacterial activity has been recognized were the cecropins . The 30- to 40-residue alpha-helical peptides display their activity by permeabilizing the membranes of bacteria . Although originally isolated from insect hemolymph, a structural and functional correlate was also found in a mammal . Here, we report on the finding that cecropin A and B from the silk moth Cecropia as well as the porcine cecropin P1 are capable of inhibiting the growth of and to kill yeast-phase Candida albicans . The peptides were tested in radial diffusion and microbroth dilution assays . They displayed potent activity against a clinical isolate as well as against defined culture strains of the pathogenic yeast but are of exceedingly low cytotoxicity towards the human cell line Jurkat . The candidacidal properties of the intensely studied molecules known to be highly active against Gram-positive and Gram-negative bacteria may renew the interest in these natural broad-spectrum peptide antibiotics and their limited cytotoxicity to human cells may be exploited for the development of topical therapeutics against pathogens resistant to classical antibiotics. Med Microbiol Immunol (Berl), 2001 Apr, 189(3), 153 - 60 Two different NO-dependent mechanisms account for the low virulence of a non-mycelial morphological mutant of Candida albicans; Diez-Orejas R et al.; We have previously described the low virulence of a Candida albicans morphological mutant: 92' . We have now used this strain to examine the role of phagocytes in its pathogenesis . Our results show that C . albicans 92' cannot evade innate host macrophage defence mechanisms as efficiently as the parental strain . In addition to the high susceptibility to phagocytosis by peritoneal macrophages, the NO produced by macrophages is a very important element in the low virulence of this agerminative mutant, a thesis supported by in vivo and in vitro experiments . Whereas the parental strain was able to inhibit macrophage NO production, the mutant was quite inefficient at reducing NO production by macrophages . In addition, the mutant showed high sensitivity to a NO generator . Treatment of mice with aminoguanidine (a preferred inducible NO synthase inhibitor) caused 90% mortality in 92' systemic infection, thus supporting a role for NO in the low virulence of this strain . Our data show that both the low inhibitory effect of 92' on macrophage NO production and the higher sensitivity to NO underlie the low virulence of this strain. Rev Prat, 2001 Apr 15, 51(7), 713 - 8 {Biology and pathogenicity of fungi}; Datry A et al.; Men live with a multitude of fungal species and few species can become parasites . Some species are obligatory pathogenic (dermatophytes, dimorphic fungi...) and the majority is opportunistic pathogen . Host-parasite complex is indissociable . For example, Candida albicans is a commensal of the gastrointestinal tract and can dramatically proliferate when host defences are alterated . Passage from the commensal status to pathogenicity is accompanied by modifications of the fungus, which then act as virulence factors (development of filamentous forms, changes in surface hydrophobicity...) . Biologic, genetic and antigenic modifications are also observed . Colonisation and adhesion, penetration, multiplication and survival are necessary for infectiosity in a host . Host defence mechanisms are described with non-specific mechanisms by humoral and cellular factors and more specific immune mechanisms involving antibodies, T lymphocytes and the role of cytokines. Allerg Immunol (Paris), 2001 Mar, 33(3), 115 - 9 {Diagnosis of normal and abnormal delayed hypersensitivity to Candida albicans . Importance of evaluating lymphocyte activation by flow cytometry}; Brunet JL et al.; Delayed type hypersensitivity (DTH) to Candida albicans is commonly observed in human . Abnormal DTH has already been described but its diagnosis is difficult to ascertain . We present now a clinical and biological study in 60 patients with a clear distinction between these two kind of Candida albicans DTH . Clinical abnormal Candida albicans DTH was characterized by a syndromic reaction 24 to 48 hours after intradermal injection . This reaction was characterized by an exacerbation of clinical symptoms . In vitro, activation of whole blood with Candida albicans antigen was detected by using flow cytometry after staining for activating markers . CD 25 positive T cells were detected in a 7 days culture in all patients . Percentage of CD 25 positive T cells was correlated to the intensity of the local cutaneous DTH reaction . CD 69 positive T cells were detected after a one day culture only in patient who presented a syndromic reaction to intradermal injection. J Immunol Methods, 2001 Jul 1, 253(1-2), 189 - 93 The use of 7-amino-actinomycin D in the analysis of Candida albicans phagocytosis and opsonization; Ortega E et al.; We describe the use of 7-amino-actinomycin D (7AAD) to measure phagocytosis and the opsonizing capacity of serum . Heat-inactivated Candida albicans was previously stained with 7AAD and incubated with resident peritoneal macrophages . The samples were analyzed by flow cytometry and phagocytic cells were identified by their bright red fluorescence . This is a rapid, reproducible and reliable one-step procedure and provides a means of evaluating low levels of phagocytosis. Yeast, 2001 Jun, 18(8), 711 - 21 Sequencing of a 4.3 kbp region of chromosome 2 of Candida albicans reveals the presence of homologues of SHE9 from Saccharomyces cerevisiae and of bacterial phosphatidylinositol-phospholipase C; Andaluz E et al.; The nucleotide sequence of a 4.3 kb fragment downstream of the LIG4 gene of Candida albicans has been determined . This fragment contains two entire ORFs (ORF1 and ORF2) and a truncated one (ORF3) . ORF1 (1029 bp; EMBL databank, Accession No . AJ277539) encodes a putative protein of 343 amino acids with a high degree of similarity to phosphatidylinositol-specific phospholipases C (PI-PLC) of bacterial origin and, to a lesser degree, to similar proteins from trypanosome, fly and human . Isolated ORF1 confers PI-PLC activity to Escherichia coli transformants . ORF2 (1572 bp; EMBL databank, Accession No . AJ277538) predicts a protein of 524 amino acids with high similarity along most of the entire length to Ydr393w from Saccharomyces cerevisiae . This protein carries a domain with significant similarity to several cytoskeleton proteins of different origins . YDR393w (SHE9) is an orphan gene whose overexpression compromises cell growth . ORF3 appears to encode the homologue of the well-conserved proteasomal 26S regulatory subunit . Crit Care Med, 2001 May, 29(5), 971 - 5 Colonization and infection of pulmonary artery catheter in cardiac surgery patients: epidemiology and multivariate analysis of risk factors; Kac G et al.; OBJECTIVE: To assess the incidence and etiology of colonization and infection of pulmonary artery catheters inserted in cardiac surgery patients . To determine the influence of some variables on the risk of developing pulmonary artery catheter colonization and infection . DESIGN: Prospective observational study of pulmonary artery catheters inserted into the internal jugular vein that were in place for >48 hrs over a 13-month period . Data collected included age, gender, nature of the cardiac surgery intervention, duration of extracorporeal circulation, date of insertion and removal, subsequent infection, and curative antimicrobial therapy . End points were pulmonary artery catheter colonization with >or=10(3) colonies on quantitative cultures and pulmonary artery catheter-related bacteremia . Risk factors for colonization were determined by multiple logistic regression . SETTING: A 17-bed cardiac surgery intensive care unit in a 480-bed teaching hospital in Paris . PATIENTS: Patients undergoing cardiac surgery procedures between May 1, 1997, and May 31, 1998 . INTERVENTIONS: None . MEASUREMENTS AND MAIN RESULTS: Of 164 pulmonary artery catheters inserted in 157 patients, 19 (11.6%) and 1 (0.6%) were associated with colonization (mean duration of catheterization, 7.5 +/- 2.8 days) and bacteremia, respectively . These data represent an incidence of 17.7 and 0.93 episodes per 1000 catheterization-days, respectively . Pulmonary artery catheter colonization was caused by Gram-positive cocci in 48% (67% were coagulase-negative staphylococci), Gram-negative rods in 48%, and Candida albicans in 4% . From multivariate analysis, >4 days of catheterization was the single variable associated with a significantly increased risk of pulmonary artery catheter colonization (odds ratio, 9.81; 95% confidence interval, 1.24-77.5, p = .03) . CONCLUSIONS: Our data show that the risk of pulmonary artery catheter-related colonization and bacteremia is quite low despite the use of a high-risk insertion site . In cardiac surgery patient populations, a trial evaluating the impact of a systematic pulmonary artery catheter removal after 4 days is warranted. FEMS Microbiol Lett, 2001 May 30, 199(2), 185 - 90 Cell cycle effects of the phenothiazines: trifluoperazine and chlorpromazine in Candida albicans; Sharma S et al.; The study demonstrates the in vitro effectiveness of phenothiazine compounds, i.e . chlorpromazine and trifluoperazine against Candida albicans . Anticandidal effect of these drugs is suggested to be because of their interaction with Ca(2+)/calmodulin dependent protein phosphorylation . 3H-thymidine uptake studies revealed that both these compounds affect the DNA synthesis along with decrease in activities of nuclear calmodulin (CaM) and Ca(2+)/calmodulin dependent protein kinase (CaMPK) . Failure in cell growth was due to defect in CaM mediated cell cycle arrest . Flow cytometric analysis showed that progression through G(1) and mitotic phase was affected when cells after alpha-factor arrest were grown in the presence of chlorpromazine or trifluoperazine . These drugs also produced significant decline in the cellular lipids and phospholipids . 14C-acetate incorporation studies further substantiated these results . We suggest that chlorpromazine or trifluoperazine affect the cell cycle through DNA synthesis (S phase) and cell division phases which are governed by calmodulin and Ca(2+)/calmodulin dependent protein phosphorylation and lipids and phospholipids appear to be additional targets of phenothiazine compounds in C . albicans . These results will have important significance in the development of new anticandidal compounds. Trends Genet, 2001 Jun, 17(6), 304 - 6 Inversions and the dynamics of eukaryotic gene order; Huynen MA et al.; Comparisons of the gene order in closely related genomes reveal a major role for inversions in the genome shuffling process . In contrast to prokaryotes, where the inversions are predominantly large, half of the inversions between Saccharomyces cerevisiae and Candida albicans appear to be small, often encompassing only a single gene . Overall the genome rearrangement rate appears higher in eukaryotes than in prokaryotes, and the current genome data do not indicate that functional constraints on the co-expression of neighboring genes have a large role in conserving eukaryotic gene order . Nevertheless, qualitatively interesting examples of conservation of gene order in eukaryotes can be observed. J Biol Chem, 2001 Jul 27, 276(30), 28413 - 20 Epub 2001 May 23. Folding requirements are different between sterol 14alpha-demethylase (CYP51) from Mycobacterium tuberculosis and human or fungal orthologs; Lepesheva GI et al.; Upon sequence alignment of CYP51 sterol 14alpha-demethylase from animals, plants, fungi, and bacteria, arginine corresponding to Arg-448 of CYP51 in Mycobacterium tuberculosis (MT) is conserved near the C terminus of all family members . In MTCYP51 Arg-448 forms a salt bridge with Asp-287, connecting beta-strand 3-2 with helix J . Deletion of the three C-terminal residues of MTCYP51 has little effect on expression of P450 in Escherichia coli . However, truncation of the fourth amino acid (Arg-448) completely abolishes P450 expression . We have investigated whether Arg-448 has other structural or functional roles in addition to folding and whether its conservation reflects conservation of a common folding pathway in the CYP51 family . Characterization of wild type protein and three mutants, R448K, R448I, and R448A, including examination of catalytic activity, secondary and tertiary structure analysis by circular dichroism and tryptophan fluorescence, and studies of both equilibrium and temporal MTCYP51 unfolding behavior, shows that Arg-448 does not play any role in P450 function or maintenance of the native structure . C-terminal truncation of Candida albicans and human CYP51 orthologs reveals that, despite conservation in sequence, the requirement for arginine at the homologous C-terminal position in folding in E . coli is not conserved . Thus, despite similar spatial folds, functionally related but evolutionarily distinct P450s can follow different folding pathways. Microbiol Res, 2001, 156(1), 95 - 102 Detection of Candida species by polymerase chain reaction (PCR) in blood samples of experimentally infected mice and patients with suspected candidemia; Khan ZU et al.; In this study, we have established and evaluated a genus-specific polymerase chain reaction (PCR) and species-specific nested PCRs for the detection of Candida species in blood samples of neutropenic mice and patients suspected of candidemia . DNA segments of the gene encoding cytochrome P450 L1A1 were targeted for amplification by using genus and species-specific primers . As compared to the genus-specific PCR, the species-specific nested PCRs improved the sensitivity by 10 times with the detection limit < 10 yeast cells . Of the 18 blood samples tested daily over a period of 8 days following Candida albicans infection in neutropenic mice, four samples were positive by genus-specific PCR and 11 were positive by species-specific nested PCR . The PCR results were correlated with culture findings obtained on blood samples . Two of the three blood culture-positive samples were positive by genus-specific PCR and all the three with species-specific nested PCR . Among 15 mice, which were negative by blood culture but had C . albicans isolated from visceral organs, 2 and 8 mice yielded positive results by genus-specific PCR and species-specific nested PCR, respectively . Consistent with the results of the animal study, species-specific nested PCR yielded much higher positivity as compared to culture (52.2% versus 21.2%) in patients suspected for candidemia . Moreover, 8 specimens which were negative for Candida by genus-specific PCR became positive by species-specific nested PCR . No correlation was apparent between PCR positivity and Candida antigen titers . The results suggest that nested PCR is a sensitive technique for the detection of Candida species from blood samples, and thus it may have application in the diagnosis of suspected cases of candidemia and candidiasis. Ann Surg, 2001 Jun, 233(6), 867 - 74 Implications of 2,457 consecutive surgical infections entering year 2000; Sawyer RG et al.; OBJECTIVE: To assess the demographics and characteristics of infections in surgical patients to define areas that deserve emphasis in surgical education . SUMMARY BACKGROUND DATA: As a result of evolving technology and diseases, the complexity of diagnosing and treating infections has increased during the past three decades for all patients, including those treated primarily by surgeons . No comprehensive analysis of these conditions in a single surgical cohort has been recently published . METHODS: The authors conducted a prospective, observational study of all infections occurring on the general and trauma surgery services at a single university hospital during a 3.5-year period . RESULTS: The authors identified 2,457 infections: 608 community-acquired, 1,053 occurring on the wards, and 796 occurring in the intensive care unit . Although dependent on patient location, the most common sites were abdomen, lung, and wound; the most common isolates were Staphylococcus epidermidis, Staphylococcus aureus, and Candida albicans; and the most commonly used antibiotics were ciprofloxacin, vancomycin, and metronidazole . The overall death rate was 13%, ranging from 5% after community-acquired infections to 25% after infections acquired in the intensive care unit . CONCLUSIONS: Most infections treated by surgeons are hospital-acquired . Infections with gram-positive cocci and fungi are common, with pulmonary infections becoming more common . Fluoroquinolones have become important therapeutic agents . Depending on the type of practice, these data should be helpful to direct educational efforts so that surgeons can remain knowledgeable and active in the nonsurgical care of their patients. J Bacteriol, 2001 Jun, 183(12), 3761 - 9 Analysis of phase-specific gene expression at the single-cell level in the white-opaque switching system of Candida albicans; Strauss A et al.; The opportunistic fungal pathogen Candida albicans can switch spontaneously and reversibly between different cell forms, a capacity that may enhance adaptation to different host niches and evasion of host defense mechanisms . Phenotypic switching has been studied intensively for the white-opaque switching system of strain WO-1 . To facilitate the molecular analysis of phenotypic switching, we have constructed homozygous ura3 mutants from strain WO-1 by targeted gene deletion . The two URA3 alleles were sequentially inactivated using the MPA(R)-flipping strategy, which is based on the selection of integrative transformants carrying a mycophenolic acid (MPA) resistance marker that is subsequently deleted again by site-specific, FLP-mediated recombination . To investigate a possible cell type-independent switching in the expression of individual phase-specific genes, two different reporter genes that allowed the analysis of gene expression at the single-cell level were integrated into the genome, using URA3 as a selection marker . Fluorescence microscopic analysis of cells in which a GFP reporter gene was placed under the control of phase-specific promoters demonstrated that the opaque-phase-specific SAP1 gene was detectably expressed only in opaque cells and that the white-phase-specific WH11 gene was detectably expressed only in white cells . When MPA(R) was used as a reporter gene, it conferred an MPA-resistant phenotype on opaque but not white cells in strains expressing it from the SAP1 promoter, which was monitored at the level of single cells by a significantly enlarged size of the corresponding colonies on MPA-containing indicator plates . Similarly, white but not opaque cells became MPA resistant when MPA(R) was placed under the control of the WH11 promoter . The analysis of these reporter strains showed that cell type-independent phase variation in the expression of the SAP1 and WH11 genes did not occur at a detectable frequency . The expression of these phase-specific genes of C . albicans in vitro, therefore, is tightly linked to the cell type. J Bacteriol, 2001 Jun, 183(12), 3582 - 8 Cloning and analysis of a Candida albicans gene that affects cell surface hydrophobicity; Singleton DR et al.; The opportunistic pathogenic yeast Candida albicans exhibits growth phase-dependent changes in cell surface hydrophobicity, which has been correlated with adhesion to host tissues . Cell wall proteins that might contribute to the cell surface hydrophobicity phenotype were released by limited glucanase digestion . These proteins were initially characterized by their rates of retention during hydrophobic interaction chromatography--high-performance liquid chromatography and used as immunogens for monoclonal antibody production . The present work describes the cloning and functional analysis of a C . albicans gene encoding a 38-kDa protein recognized by the monoclonal antibody 6C5-H4CA . The 6C5-H4CA antigen was resolved by two-dimensional electrophoresis, and a partial protein sequence was determined by mass spectrometry analysis of tryptic fragments . The obtained peptides were used to identify the gene sequence from the unannotated C . albicans DNA database . The antibody epitope was provisionally mapped by peptide display panning, and a peptide sequence matching the epitope was identified in the gene sequence . The gene sequence encodes a novel open reading frame (ORF) of unknown function that is highly similar to several other C . albicans ORFs and to a single Saccharomyces cerevisiae ORF . Knockout of the gene resulted in a decrease in measurable cell surface hydrophobicity and in adhesion of C . albicans to fibronectin . The results suggest that the 38-kDa protein is a hydrophobic surface protein that meditates binding to host target proteins. Biochemistry, 2001 May 29, 40(21), 6520 - 6 Binding enhancement by tertiary interactions and suicide inhibition of a Candida albicans group I intron by phosphoramidate and 2'-O-methyl hexanucleotides; Disney MD et al.; Candida albicans is one of many infectious pathogens that are evolving resistance to current treatments . RNAs provide a large class of targets for new therapeutics for fighting these organisms . One strategy for targeting RNAs uses short oligonucleotides that exhibit binding enhancement by tertiary interactions in addition to Watson-Crick pairing . A potential RNA target in C . albicans is the self-splicing group I intron in the LSU rRNA precursor . The recognition elements that align the 5' exon splice site for a ribozyme derived from this precursor are complex {Disney, M . D., Haidaris, C . G., and Turner, D . H . (2001) Biochemistry 40, 6507-6519} . These recognition elements have been used to guide design of hexanucleotide mimics of the 5' exon that have backbones modified for nuclease stability . These hexanucleotides bind as much as 100000-fold more tightly to a ribozyme derived from the intron than to a hexanucleotide mimic of the intron's internal guide sequence, r(GGAGGC) . Several of these oligonucleotides inhibit precursor self-splicing via a suicide inhibition mechanism . The most promising suicide inhibitor is the ribophosphoramidate rn(GCCUC)rU, which forms more trans-spliced than cis-spliced product at oligonucleotide concentrations of >100 nM at 1 mM Mg(2+) . The results indicate that short oligonucleotides modified for nuclease stability can target catalytic RNAs when the elements of tertiary interactions are complex. Biochemistry, 2001 May 29, 40(21), 6507 - 19 Recognition elements for 5' exon substrate binding to the Candida albicans group I intron; Disney MD et al.; A group I intron precursor and ribozyme were cloned from the large subunit rRNA of the human pathogen Candida albicans . Both the precursor and ribozyme are functional as determined from in vitro assays . Comparisons of dissociation constants for oligonucleotide binding to the ribozyme and to a hexanucleotide mimic of its internal guide sequence lead to a model for recognition of the 5' exon substrate by this intron . In particular, tertiary contacts with the P1 helix that help align the splice site include three 2'-hydroxyl groups, a G.U pair that occurs at the intron's splice junction, and a G.A pair . The free energy contribution that each interaction contributes to tertiary binding is determined . When the G.A pair is replaced with a G-C pair, tertiary interactions to 5' exon mimic 2'-hydroxyl groups are significantly weakened . When the G.A pair is replaced with a G.U pair, tertiary interactions are retained and binding is 10-fold tighter . These results expand our knowledge of substrate recognition by group I introns, and also provide a basis for rational design of oligonucleotide-based therapeutics for targeting group I introns by binding enhancement by tertiary interactions and suicide inhibition strategies. Mol Genet Genomics, 2001 Mar, 265(1), 172 - 9 Cla4 protein kinase is essential for filament formation and invasive growth of Yarrowia lipolytica; Szabo R; The non-conventional yeast Yarrowia lipolytica is a suitable model for the study of yeast dimorphism . In order to identify genes that may be involved in the regulation of this process, random mutagenesis was performed . This led to the isolation of monomorphic mutants that had lost the ability to grow in a hyphal form both in liquid and on solid medium . Filamentation was restored to one of the mutants by transformation with a fragment of Y . lipolytica genomic DNA containing a single 2766-bp ORF . The predicted protein has a molecular weight of 99.6 kDa and is highly homologous to the protein kinases Cla4 of Candida albicans and Saccharomyces cerevisiae, which are members of the p21-activated kinase (PAK) family . Analysis of the putative protein sequence identified conserved C-terminal catalytic, and internal Cdc42p-binding regions, as well as a pleckstrin homology domain typical of PAK kinases . The results indicate that CLA4 is a single-copy gene located on the chromosome V of Y . lipolytica . Deletion of CLA4 is not lethal, but completely eliminates the ability to form filaments and to invade agar . A strain lacking a functional CLA4 gene exhibits an aberrant distribution of chitin in the cell wall, indicating a possible role for the Cla4 protein kinase in the maintenance of cell polarity in Y . lipolytica. Saudi Med J, 2000 Oct, 21(10), 928 - 30 Otitis externa in a localized area at the South of Jordan; Al-Asaaf SM et al.; OBJECTIVE: To investigate the clinical findings and to identify the microorganisms in the pathogenesis of otitis externa . METHODS: The symptoms and signs, as well as the bacterial flora, from the auditory canals of 70 Jordanian patients (94 ears) suffering from otitis externa were studied during the period from February 1999 to February 2000 . RESULTS: Pain was the most common symptom (82%) . The most common sign was erythema (65%), while oedema was the least common (42%) . Pseudomonas aeruginosa was found in 39%, Aspergillus in 27%, Candida albicans in 18%, Staphylococcus aureus in 18% and no growth in 8.5% . Ninety three percent of healthy ears revealed normal skin flora . CONCLUSION: Gram-negative microorganisms (Pseudomonas aeruginosa) were the most frequently isolated pathogenic microorganisms. Biochem J, 2001 Jun 1, 356(Pt 2), 595 - 604 Characterization of the gene family encoding alternative oxidase from Candida albicans; Huh WK et al.; Candida albicans possesses a cyanide-resistant respiratory pathway mediated by alternative oxidase (AOX), which seems to be encoded by a gene family with two members . Cloning and expression of AOX1a, one of the genes encoding alternative oxidase from C . albicans, has previously been reported {Huh and Kang (1999) J . Bacteriol . 181, 4098-4102} . In the present study we report the isolation of another gene coding for alternative oxidase, designated AOX1b . AOX1b contains a continuous open reading frame that encodes a polypeptide consisting of 365 amino acids . Interestingly, AOX1a and AOX1b were found to be located in tandem on one of the chromosomes of C . albicans . The presence of cyanide in the culture medium remarkably retarded the growth of the aox1a/aox1a mutants . The growth of the aox1b/aox1b mutants and the aox1a/aox1a aox1b/aox1b double mutants was almost completely inhibited in the same medium . beta-Galactosidase reporter assays indicated that, whereas AOX1a was expressed constitutively, the expression of AOX1b was dependent on growth phase and was induced by treatment with cyanide, antimycin A, H(2)O(2), menadione and paraquat . Growth of the cells in media with non-fermentable carbon sources also enhanced the expression of AOX1b . CaSLN1, which encodes a histidine kinase, seems to be involved in the regulation of AOX expression in C . albicans on the basis of the observation that the activity of cyanide-resistant respiration and the expression level of AOX in the casln1/casln1 mutants were found to be significantly low under normal conditions and slightly increased in the presence of respiratory inhibitors compared with the wild-type strain . Like AOX1a, AOX1b could also be functionally expressed in AOX-deficient Saccharomyces cerevisiae and confer cyanide-resistant respiration on the organism. Biochem J, 2001 Jun 1, 356(Pt 2), 361 - 8 Effects of histatin 5 and derived peptides on Candida albicans; Ruissen AL et al.; Three anti-microbial peptides were compared with respect to their killing activity against Candida albicans and their ability to disturb its cellular and internal membranes . Histatin 5 is an anti-fungal peptide occurring naturally in human saliva, while dhvar4 and dhvar5 are variants of its active domain, with increased anti-microbial activity . dhvar4 has increased amphipathicity compared with histatin 5, whereas dhvar5 has amphipathicity comparable with that of histatin 5 . All three peptides caused depolarization of the cytoplasmic and/or mitochondrial membrane, indicating membranolytic activity . For the variant peptides both depolarization and killing occurred at a faster rate . With FITC-labelled peptides, no association with the cytoplasmic membrane was observed, contradicting the formation of permanent transmembrane multimeric peptide pores . Instead, the peptides were internalized and act on internal membranes, as demonstrated with mitochondrion- and vacuole-specific markers . In comparison with histatin 5, the variant peptides showed a more destructive effect on mitochondria . Entry of the peptides and subsequent killing were dependent on the metabolic state of the cells . Blocking of the mitochondrial activity led to complete protection against histatin 5 activity, whereas that of dhvar4 was hardly affected and that of dhvar5 was affected only intermediately. Pharmacogenomics, 2001 May, 2(2), 95 - 106 Regulation of both gene expression and protein stability provides genetically assisted target evaluation (GATE) for microbial target validation; Averett DR et al.; The attempt to develop novel antibiotics, active against organisms resistant to current therapies, has led researchers to seek and explore new drug targets . The rapid sequencing and analysis of entire microbial genomes has identified large numbers of genes that may be sufficiently different from their human counterparts to be exploited as targets for antimicrobial treatment . As a first step, the importance of the various putative targets for microbial growth and survival must be assessed . Emerging validation technologies are becoming increasingly sophisticated and, in certain cases, allow prioritisation of the best targets . In this paper, genetically assisted target evaluation (GATE) is introduced as a versatile target validation technology . GATE concomitantly manipulates both synthesis and stability of the targeted protein using copper ions as an effector . This technology allows rapid quantitation of the lethal consequences of inactivation of targeted gene products in Saccharomyces cerevisiae . Additional tools can then be applied to extend these results into pathogenic organisms, such as Candida albicans. J Agric Food Chem, 2001 May, 49(5), 2327 - 32 Identification and biological activities of triterpenoid saponins from Chenopodium quinoa; Woldemichael GM et al.; At least 16 saponins were detected in the seeds of Chenopodium quinoa . The 5 previously isolated major saponins, 3-O-beta-D-glucuronopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L-arabinopyranosyl phytolaccagenic acid 28-O-beta-D-glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl phytolaccagenic acid 28-O-beta-D-glucopyranosyl ester, and the new saponin 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl phytolaccagenic acid were isolated and characterized using mainly NMR spectroscopy, mass spectrometry, and chemical methods . The antifungal activity against Candida albicans and hemolytic activity on erythrocytes of these compounds and derived monodesmosides were evaluated . Both bidesmosides and derived monodesmosides showed little or no antifungal activity, whereas a comparatively higher degree of hemolytic activity could be determined for monodesmosides. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 1997 Dec, 14(4), 373 - 5 {Primary study of bacterial adhesion of prosthetic valve materials}; Zhang E et al.; The adhesion of Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans to Dacron and pyrolite carbon was measured in vitro with plate counting . The experimental result showed: the bacterials used in the experiment have adhesion to prosthetic valve materials; the bacterials have stronger adhesion to Dacron than to Pyrolite carbon; the bacterial adhesive capacity is significantly different and is affected by mechanical and chemical nature of biomaterials; Staphylococcur aureus has the strongest adhesion to Dacron, and Pseudomonas aeruginosa has the strongest adhesion to pyrolite carbon in all bacterials. Int Immunopharmacol, 2001 Jan, 1(1), 167 - 75 Stimulation of nonspecific immunity, haemopoiesis and protection of mice against radiation injury by 1-adamantylamide-L-alanyl-D-isoglutamine incorporated in liposomes; Turanek J et al.; 1-Adamantylamide-L-alanyl-D-isoglutamine (adamantylamide dipeptide (AdDP)) belongs to a group of desmuramyl muramyl peptide derivatives which are able to protect an organism from some viral infections . Encapsulation of AdDP to egg phosphatidyl choline liposomes and the targeting of this drug to lymphatic node macrophages via subcutaneous (s.c.) administration proved to be the efficient way to protect mice against irradiation when administered s.c., 24 h prior to lethal gamma-irradiation (long-term survival rate in the range of 40% compared with 0% in saline or free drug control) . Parameters characteristic for the recovery of haemopoiesis in the bone marrow (number of granulocyte-macrophage haemopoietic progenitor cells, granulocyte-macrophage colony forming cells (GM-CFC)) were significantly improved in comparison with the controls and free drug on day 10 after 6.5 Gy irradiation . The haemopoietic effect was observed in the broad application time window (72 h before and 48 h after irradiation) . Very high radioprotective effect of s.c . administered liposomal AdDP (L-AdDP) can be explained (together with induction of haemopoiesis) by the effective and long-lasting activation of nonspecific immunity, which withholds the onset of septicemia in early days after irradiation . Induction of nonspecific immunity was proven in Candida albicans infectious model . L-AdDP significantly increased both the survival time and score (about 40% survival compared with 0% in controls and free drug) . In conclusion, L-AdDP could be therapeutically beneficial to moderate the haemopoietic damage (undesirable effect of radiotherapy or chemotherapy) and induce the non-specific immunity to support the antimicrobial treatment of immunocompromised patients. AIDS Clin Care, 1995 Mar, 7(3), 19 - 22 Management of HIV-associated esophageal disease; Belitsos PC; AIDS: Esophageal conditions due to fungal, ulcerative, and neoplastic causes often signal the onset of symptomatic HIV infection . Most cases are fungal and due to Candida albicans, which is characterized by esophageal inflammation causing pain on swallowing (dysphagia and odynophagia) . Ulcerative esophageal disease is commonly associated with cytomegalovirus (CMV), idiopathic causes, and herpes simplex virus (HSV) . CMV, characterized by odynophagia resulting from ulcerations in the distal third of the esophagus, is clinically indistinguishable from idiopathic ulceration . HSV is more widespread and abrupt than other ulcerative processes, and its erosive injury can cause painful swallowing, ulceration and oral cavity lesions . Patients with esophageal distress, low CD4 counts, and little possibility of other GI conditions most likely suffer from Candida infection and should immediately begin an empiric trial of antifungal therapy . If an individual's first bout of odynophagia does not respond to empiric oral azole therapy, the diagnosis of fungal esophagitis is probably incorrect and an upper endoscopic evaluation should be performed . Patients generally respond quickly and completely to treatment of a first episode of fungal esophagitis; therefore, neither primary prophylaxis nor long-term suppressive therapy are recommended due to the risk of infection with a resistant strain . Failure of patients on suppressive therapy to respond to antifungal medication usually indicates resistant fungal infection that may require treatment with intravenous amphotericin . If CMV-isolated esophagitis is diagnosed, the patient should begin intravenous ganciclovir, followed by IV foscarnet if the healing after three weeks is minimal . AIDS Patient Care STDS, 1998 May, 12(5), 359 - 66 Vaginal candidiasis: review and role of local mucosal immunity; Fidel PL Jr; Vulvovaginal candidiasis is a common mucosal fungal infection in women of child-bearing ages . Despite the role for cell-mediated immunity and T cells in host protection against the majority of mucosal Candida albicans infections, there is controversy as to whether immunosuppression by HIV infection enhances susceptibility to vaginal candidiasis . To date, host defense against C . albicans vaginitis has been studied in women with recurrent vaginitis, in HIV-infected women, and in animal models of experimental vaginitis . Together, data suggest that local immunity is more important than that in the systemic circulation for host defense against vaginitis . Thus, current investigations have been focused specifically on innate and acquired immune responses against C . albicans at the vaginal mucosa . This review will discuss the current knowledge of host defenses against C . albicans vaginitis, both systemically and locally, and try to shed some light on several issues surrounding the efficiency of this seemingly compartmentalized immune response. Pediatr Dermatol, 2001 Mar-Apr, 18(2), 149 - 55 Diaper dermatitis: a therapeutic dilemma . Results of a double-blind placebo controlled trial of miconazole nitrate 0.25%; Concannon P et al.; Diaper dermatitis, an acute inflammation of the skin in the diaper area, is the most common dermatologic disorder of infancy . This placebo-controlled, randomized, double-blind, parallel-group trial compared the efficacy and safety of miconazole nitrate 0.25% in a zinc oxide/petrolatum base with that of the ointment base alone in treating acute diaper dermatitis in infants and evaluated the role of Candida albicans in the response to treatment . Infants age 2-13 months with diaper rash were treated with either miconazole nitrate 0.25% (N = 101) or ointment base (N = 101) for 7 days . Although improvement in rash from baseline was seen in both treatment groups on days 3, 5, and 7, patients receiving miconazole nitrate 0.25% had significantly fewer rash sites and lower mean total rash scores on days 5 and 7 (p < 0.001) . In the miconazole nitrate 0.25% group, improvement was most marked among those with moderate or severe diaper dermatitis at baseline and among patients whose baseline rashes were positive for C . albicans . Treatment with miconazole nitrate 0.25% was as safe as with ointment base alone . Miconazole nitrate 0.25% ointment is a safe and effective treatment for diaper dermatitis in infants. Oral Dis, 2001 Mar, 7(2), 119 - 22 The impact of chlorhexidine gluconate on the relative cell surface hydrophobicity of oral Candida albicans; Anil S et al.; OBJECTIVES: Adherence of Candida albicans has been implicated as the first step in the pathogenesis of oral candidosis, and its relative cell surface hydrophobicity (CSH) a contributory physical force . Chlorhexidine gluconate is by far the commonest antiseptic mouthwash prescribed in dentistry . The intra-oral concentrations of the retained chlorhexidine mouthwash fluctuate considerably due to the dilution effect of saliva and the cleansing action of the oral musculature . Hence the objective of the present study was to investigate the effect of brief exposure to sub-therapeutic concentrations of chlorhexidine gluconate on the relative CSH of C . albicans . DESIGN: The CSH of the isolates was assessed by a biphasic aqueous-hydrocarbon assay . RESULTS: A statistically significant reduction in CSH was observed following the exposure of Candida isolates to 0.005 and 0.0025% chlorhexidine gluconate . CONCLUSIONS: These results elucidate additional mechanisms by which chlorhexidine gluconate suppress candidal pathogenicity despite a brief period of transient exposure within the oral environment. Oral Dis, 2001 Jan, 7(1), 11 - 7 Adjunctive use of chlorhexidine in oral candidoses: a review; Ellepola AN et al.; Oral candidosis is by far the commonest human fungal infection and manifests in a variety of clinical guises . The main reason for its high incidence appears to be the multiplicity of predisposing factors, which facilitate the conversion of oral commensal Candida to a parasitic existence . Despite the availability of a number of effective antimycotics for the treatment of oral candidoses, failure of therapy is not uncommon owing to the unique environment of the oral cavity where the flushing effect of saliva and the cleansing action of the oral musculature tend to reduce the drug concentration to sub-therapeutic levels . For these and other reasons chlorhexidine is widely prescribed in dentistry both as an antiseptic mouthwash and a denture disinfectant in order to supplement other antifungals . Chlorhexidine has a broad spectrum of antimicrobial activity including Candida albicans and other common non-albicans yeast species . In this review we outline the utility of chlorhexidine as an adjunct to conventional antimycotic therapy in the management of oral Candida infections. Indian J Med Sci, 2000 Nov, 54(11), 491 - 4 Bronchopulmonary candidiasis in a tertiary referral hospital of Assam, India; Phukan AC et al.; One hundred patients with chronic chest infection suffering for more than three months admitted into a tertiary referral Hospital, northeast India were examined for pulmonary mycoses . The morning sputum samples in 3 consecutive days with a throat swab of each patient were examined for detection, isolation and identification of the fungus . Study showed Pulmonary candidiasis in 50% of the patients where Candida albicans were having highest incidence of association followed by 5 other species of Candida . Pre-existing conditions like pulmonary tuberculosis, bronchogenic carcinoma, lung abscess, bronchial asthma make the lungs prone to be invaded by the candida species . Long term antibiotics and steroids therapy was found to be associated with pulmonary candidiasis . Other conditions like irradiation treatment, malignancy, diabetes mellitus and malnutrition were also found to be the predisposing factors which influence bronchopulmonary candidiasis. Bioorg Med Chem, 2001 Apr, 9(4), 931 - 8 Amide and ester derivatives of N3-(4-methoxyfumaroyl)-(S)-2,3-diaminopropanoic acid: the selective inhibitor of glucosamine-6-phosphate synthase; Zgodka D et al.; Several amide and ester derivatives of a glutamine analogue, N3-(4-methoxyfumaroyl)-(S)-2,3-diaminopropanoic acid (FMDP) (1-8), were synthesized and evaluated for the inhibitory activity in regard to glucosamine-6-phosphate synthase from Candida albicans . The syntheses were accomplished by the reaction of N2-tert-butoxycarbonyl-N3-(4-methoxyfumaroyl)-(S)-2,3-diaminopropanoic acid (BocFMDP) with the corresponding amines to give the FMDP amides (1-4) or with alkyl halides to give corresponding esters of FMDP (5-8) . Among the synthesized compounds, the acetoxymethyl ester of FMDP was the most active inhibitor of the enzyme . Its IC50 value compared to that of FMDP (4 microM) was equal to 11.5 microM . The methyl and allyl esters and the N-hexyl-N-methyl-amide of FMDP exhibited a moderate enzyme inhibitory activity. Antimicrob Agents Chemother, 2001 Jun, 45(6), 1660 - 70 Genomic profiling of the response of Candida albicans to itraconazole treatment using a DNA microarray; De Backer MD et al.; The application of genome-wide expression profiling to determine how drugs achieve their therapeutic effect has provided the pharmaceutical industry with an exciting new tool for drug mode-of-action studies . We used DNA chip technology to study cellular responses to perturbations of ergosterol biosynthesis caused by the broad-spectrum antifungal agent itraconazole . Simultaneous examination of over 6,600 Candida albicans gene transcript levels, representing the entire genome, upon treatment of cells with 10 microM itraconazole revealed that 296 genes were responsive . For 116 genes transcript levels were decreased at least 2.5-fold, while for 180 transcript levels were similarly increased . A global upregulation of ERG genes in response to azole treatment was observed . ERG11 and ERG5 were found to be upregulated approximately 12-fold . In addition, a significant upregulation was observed for ERG6, ERG1, ERG3, ERG4, ERG10, ERG9, ERG26, ERG25, ERG2, IDII, HMGS, NCP1, and FEN2, all of which are genes known to be involved in ergosterol biosynthesis . The effects of itraconazole on a wide variety of known metabolic processes are discussed . As over 140 proteins with unknown function were responsive to itraconazole, our analysis might provide-in combination with phenotypic data-first hints of their potential function . The present report is the first to describe the application of DNA chip technology to study the response of a major human fungal pathogen to drug treatment. Infect Immun, 2001 Jun, 69(6), 3939 - 46 Deficiency of D-erythroascorbic acid attenuates hyphal growth and virulence of Candida albicans; Huh WK et al.; In some lower eukaryotes, D-erythroascorbic acid, a five-carbon analog of L-ascorbic acid, is present instead of L-ascorbic acid . We have cloned ALO1, the gene encoding D-arabinono-1,4-lactone oxidase, which catalyzes the final step of D-erythroascorbic acid biosynthesis in Candida albicans . The ALO1 gene contained a continuous open reading frame of 1,671 bp that encodes a polypeptide consisting of 557 amino acids with a calculated molecular mass of 63,428 Da . To investigate the functional roles of D-erythroascorbic acid in C . albicans, we disrupted or overexpressed the ALO1 gene . In the alo1/alo1 null mutants, the activity of D-arabinono-1,4-lactone oxidase was completely lost and D-erythroascorbic acid could not be detected . When ALO1 on a multicopy plasmid was transformed in C . albicans, the enzyme activity and the intracellular D-erythroascorbic acid level were increased up to 3.4-fold and 4.0-fold, respectively . The alo1/alo1 null mutants of C . albicans showed increased sensitivity towards oxidative stress . Overexpression of ALO1 made the cells more resistant to the same stress . The alo1/alo1 mutants showed defective hyphal growth and attenuated virulence . Taken together, our results suggest that D-erythroascorbic acid functions as an important antioxidant and can be considered one of the virulence factors enhancing the pathogenicity of C . albicans. Infect Immun, 2001 Jun, 69(6), 3883 - 90 Lactoferrin peptide increases the survival of Candida albicans-inoculated mice by upregulating neutrophil and macrophage functions, especially in combination with amphotericin B and granulocyte-macrophage colony-stimulating factor; Tanida T et al.; To develop a new strategy to control candidiasis, we examined in vivo the anticandidal effects of a synthetic lactoferrin peptide, FKCRRWQWRM (peptide 2) and the peptide that mimics it, FKARRWQWRM (peptide 2') . Although all mice that underwent intraperitoneal injection of 5 x 10(8) Candida cells with or without peptide 2' died within 8 or 7 days, respectively, the survival times of mice treated with 5 to 100 microg of intravenous peptide 2 per day for 5 days after the candidal inoculation were prolonged between 8.4 +/- 2.9 and 22.4 +/- 3.6 days, depending on the dose of peptide 2 . The prolongation of survival by peptide 2 was also observed in mice that were infected with 1.0 x 10(9) Candida albicans cells (3.2 +/- 1.3 days in control mice versus 8.2 +/- 2.4 days in the mice injected with 10 microg of peptide 2 per day) . In the high-dose inoculation, a combination of peptide 2 (10 microg/day) with amphotericin B (0.1 microg/day) and granulocyte-macrophage colony-stimulating factor (GM-CSF) (0.1 microg/day) brought prolonged survival . With a combination of these agents, 60% of the mice were alive for more than 22 days . Correspondingly, peptide 2 activated phagocytes inducing inducible NO synthase and the expression of p47(phox) and p67(phox), and peptide 2 increased phagocyte Candida-killing activities up to 1.5-fold of the control levels upregulating the generation of superoxide, lactoferrin, and defensin from neutrophils and macrophages . These findings indicated that the anticandidal effects of peptide 2 depend not only on the direct Candida cell growth-inhibitory activity, but also on the phagocytes' upregulatory activity, and that combinations of peptide 2 with GM-CSF and antifungal drugs will help in the development of new strategies for control of candidiasis. Infect Immun, 2001 Jun, 69(6), 3728 - 36 Antigenic properties and processing requirements of 65-kilodalton mannoprotein, a major antigen target of anti-Candida human T-cell response, as disclosed by specific human T-cell clones; Nisini R et al.; T-cell-mediated immunity is known to play a central role in the host response to Candida albicans . T-cell clones are useful tools for the exact identification of fungal T-cell epitopes and the processing requirements of C . albicans antigens . We isolated human T-cell clones from an HLA-DRB1*1101 healthy donor by using an antigenic extract (MP-F2) of the fungus . Specific clones were T-cell receptor alpha/beta and CD4(+)/CD8(-) and showed a T-helper type 1 cytokine profile (production of gamma interferon and not interleukin-4) . The large majority of these clones recognized both the natural (highly glycosylated) and the recombinant (nonglycosylated) 65-kDa mannoprotein (MP65), an MP-F2 minor constituent that was confirmed to be an immunodominant antigen of the human T-cell response . Surprisingly, most of the clones recognized two synthetic peptides of different MP65 regions . However, the peptides shared the amino acid motif IXSXIXXL, which may be envisaged as a motif sequence representing the minimal epitope recognized by these clones . Three clones recognized natural and pronase-treated MP65 but did not detect nonglycosylated, recombinant MP65 or the peptides, suggesting a possible role for polysaccharides in T-cell recognition of C . albicans . Finally, lymphoblastoid B-cell lines were efficient antigen-presenting cells (APC) for recombinant MP65 and peptides but failed to present natural, glycosylated antigens, suggesting that nonprofessional APC might be defective in processing highly glycosylated yeast proteins . In conclusion, this study provides the first characterization of C . albicans-specific human T-cell clones and provides new clues for the definition of the cellular immune response against C . albicans. Farmaco, 2001 Jan-Feb, 56(1-2), 81 - 5 Microbial fermentation-derived inhibitors of efflux-pump-mediated drug resistance; Lee MD et al.; A library of 85000 microbial fermentation extracts was screened for inhibitors of multidrug resistance efflux pumps in Pseudomonas aeruginosa and Candida albicans . New compounds EA-371alpha and EA-371delta were isolated and demonstrated to be potent and specific inhibitors of the MexAB-OprM pump in P . aeruginosa . Two series of fungal metabolites, enniatins and beauvericins, were found to be ubiquitous and potent inhibitors of ABC transporters . Milbemycins were rediscovered as potent inhibitors of the CDRI pump in C . albicans, and demonstrated to potentiate effectively the antifungal activity of fluconazole and SCH-56592 against a wide variety of Candida clinical isolates. Biomed Sci Instrum, 2001, 37, 167 - 72 An evaluation of infectious diseases in cervicovaginal smears from patients with atypical cells of undetermined significance; Coach S et al.; Despite all studies of infectious disease of the female genital tract, there have been a few studies of the many different types of infectious organisms on Pap smears that contain abnormal cells . The purpose of this study is to evaluate the significance of infectious organisms in women with a cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS) . This study utilizes 398 cervicovaginal smears and concurrent cervical biopsies (test and control cases) obtained from the Department of Health cases from January to December 1993 and 1995 . The cases were randomized, assigned a study number and reviewed for the presence of infectious organisms . The age range for the patients consisted of 15 to 62 for the ASCUS test group (mean 24) and 13 to 64 for the matched control group (mean 24) . The results of the microscopic review were entered into SPSS for evaluation . Infectious organisms were identified in 101 (25.4%) of the test and control cases . The type of organisms identified in the ASCUS and matched control cases were normal vaginal flora 42 (21.2) and 48 (24.0%), Actinomyces one (.3%) case, Candida albicans in 2 (1%) and 4 (2%), Trichomonas vaginalis in 10 (5.1%) and 9 (4.5%) . Herpes virus effect was not identified in the test cases but was identified in 2 of the control cases . The cytopathic effect of human papilloma virus was identified in 68 (34.3) and 3 (1.5%) of the test cases . Data collected in this study indicate that infectious organisms, other than the effect of the human papilloma virus, are not as common in the ASCUS cases as the matched control cases . Human papilloma virus effect was more commonly associated with the squamous intraepithelial lesions. Folia Microbiol (Praha), 2000, 45(4), 325 - 9 Effect of peroxynitrite on dormant spores and germlings of Aspergillus fumigatus in vitro; Kunert J; Peroxynitrite was tested for its effects on the opportunistic pathogenic fungus Aspergillus fumigatus . It did not kill any dormant or swollen (4 h in a glucose-peptone medium) conidia in concentrations up to 6.25 mmol/L and the growth of germlings (after 6 or 9 h) was only slightly inhibited by 5 mmol/L peroxynitrite . The peroxynitrite donor SIN-1 (up to 10 mmol/L, 1 d in buffer) did not kill any conidia but inhibited their germination and growth, depending on the medium . Ten mmol/L SIN-1 in a poor medium was fungistatic and germination was stopped for 20 h . Nine strains of A . fumigatus showed resistance comparable to the model strain, while 6 Candida albicans strains were much more susceptible to both peroxynitrite and its donor . The results indicate that peroxynitrite does not contribute substantially to the antifungal activity of phagocytes against A . fumigatus. Folia Microbiol (Praha), 2000, 45(4), 313 - 9 Influence of methylfenpropidine on growth, sterol content and fatty acid composition of Candida albicans; Sajbidor J et al.; The effect of methylfenpropidine on growth, lipid contents, sterol and fatty acid composition was investigated in 5 strains of Candida albicans . The sensitivity of the strains decreased in the order: wild strains > erg+ ade nysR > ade nysR erg (defective delta (8-7)-isomerase) > ade nysR erg (defective delta 5-desaturase) . The presence of the inhibitor influenced fecosterol isomerization, episterol dehydrogenation, zymosterol transmethylation, ignosterol reduction and squalene epoxidation . Methylfenpropidine also induced changes in fatty acid composition, causing a reduction of the palmitic and oleic acid content with a concomitant elevation of stearic, linoleic and linolenic acid levels . The lipid unsaturation index slightly increased . Morphological changes of wild strains were observed after the fungicide treatment. Bull Soc Pathol Exot, 2001 Mar, 94(1), 11 - 3 {Ocular parasitoses and mycoses: cases diagnosed in the Central University Hospital of Sfax between 1996 and 1999}; Cheikh-Rouhou F et al.; Parasitical and fungal ophthalmic infections are polymorphic and variably severe . They are rarely reported in publications . The aim of our study has been to specify the parasitic and fungal agents isolated from the ocular samples in our district (Sfax-Tunisia) . We surveyed retrospectively the different ocular parasitosis and mycosis diagnosed in our laboratory (Sfax hospital) over a 4 year-period (1996-1999) . Fungal ophthalmic infections were dominated by corneal localisations: 20 cases of keratomycosis secondary to: Fusarium solani (8 cases), Aspergillus fumigatus (3 cases); Aspergillus flavus (2 cases); Alternaria sp (2 cases), Candida albicans (2 cases); Fusarium dimerium (1 case); Fusarium oxysporum (1 case) and Scedosporium sp (1 case) . A prolonged treatment by ketoconazole had a successful resolution in 70% of cases; 1 case of ciliar tinea caused by Trichophyton violaceum . Parasitic agents were dominated by Demodex folliculorum (32 cases), Phthirius inguinalis (6 cases) and Oestrus ovis (2 cases) . Two cases of orbital hydatidosis and 2 cases of palpebral cutaneous leishmania were noted; 21 cases of ocular toxoplasmosis were treated by clindamycine . Our survey concerned not only cosmopolite parasitosis and fungi, but also some affections endemic to our district (hydatidosis and leishmaniasis) with manifestations in isolated ophthalmic localisation . Our research has underscored the need to specify types of infection by way of adequate sampling so as to treat early and then improve prognosis. Med Mycol, 2001 Apr, 39(2), 207 - 13 DNA fingerprinting of serial Candida albicans isolates obtained during itraconazole prophylaxis in patients with AIDS; Le Monte AM et al.; During a randomized double-blind placebo-controlled study testing the efficacy of itraconazole for prophylaxis of systemic and mucosal fungal infections in patients with acquired immune deficiency syndrome, 298 patients were enrolled with 295 evaluable . Of those, 46 patients were considered prophylaxis failures because of recurrent oral or esophageal candidiasis . Oropharyngeal fungal cultures were taken at the time of suspected thrush or Candida esophagitis, but not at baseline . All of the Candida spp . isolates were cultured on CHROMagar Candida medium then identified using API 20 AUX strips . Antifungal susceptibility testing was performed following the National Committee for Clinical Laboratory Standards M-27A guidelines . Sequential isolates were genotyped using randomly amplified polymorphic DNA . Polymerase chain reaction fingerprints were generated using two repetitive sequence primers, (GGA)7 and (GACA)4 . The study group consisted of 23 patients, nine from the itraconazole arm and 14 from the placebo arm, who were prophylaxis failures and had more than two C . albicans isolates . Five of 23 had isolates showing a > or =4-fold reduction in susceptibility; four of these patients were in the itraconazole prophylaxis arm and one was in the placebo arm . Three of the five had yeast isolations showing changes in banding patterns over time . Such changes may indicate genetic changes in the same strain that could be linked to acquired resistance to itraconazole, or acquisition of a new strain, or emergence of a previously minor component of the original population. Med Mycol, 2001 Apr, 39(2), 185 - 93 PCR fingerprinting: a convenient molecular tool to distinguish between Candida dubliniensis and Candida albicans; Meyer W et al.; Candida dubliniensis was recently identified as a germ-tube- and chlamydospore-positive yeast, phenotypically and morphologically indistinguishable from the phylogenetically closely related yeast species C . albicans and its synonymized variant C . stellatoidea . The high similarity between these yeast species causes significant problems in the correct identification of C . dubliniensis in a standard clinical mycology laboratory . Polymerase chain reaction (PCR) fingerprinting was successfully applied here to distinguish between clinical isolates of the two closely related species . Microsatellite ({GACA}4) and minisatellite ({5'-GAGGGTGGCGGTTCT-3'}, derived from the core-sequence of the wild-type phage M13) specific oligonucleotides were used as single primers in PCR to amplify hypervariable inter-repeat DNA sequences from 16 C . dubliniensis strains and 11 C . albicans strains . Each species, represented by its ex-type strain, could be identified by a distinct species-specific multilocus pattern, allowing identification to species level for all clinical isolates . In addition, the PCR fingerprinting generated strain-specific profiles, making this method applicable to epidemiological investigations . PCR fingerprinting using the primer M13 is proposed here as a simple, reliable and highly reproducible molecular tool to differentiate between strains of C . albicans and C . dubliniensis. Microbiol Immunol, 2001, 45(3), 201 - 8 Effect of sealing and Tween 80 on the antifungal susceptibility testing of essential oils; Inouye S et al.; Concentrations of essential oils showing high volatility decreased substantially in broth and agar media when incubated under open conditions . The decrease in the half life was from 0.7 to 38 hr in broth medium at 27 C . When evaporation was prevented by sealing, MIC values against Aspergillus fumigatus, Candida albicans and Trichophyton mentagrophytes by broth or agar dilution assay were lowered two to eight-fold, as compared with those obtained under open conditions . Addition of Tween 80 caused a rise of the MICs against A . fumigatus by two to four-fold in broth dilution assay, but little affected the MICs in agar dilution assay. Scand J Infect Dis, 2001, 33(4), 312 - 3 Mitral valve endocarditis following balloon mitral valvotomy; Sood P et al.; The case of a patient who developed fungal valve endocarditis due to Candida albicans following balloon mitral valvotomy is presented . The patient did not have any obvious predisposing factors which led to the development of fungal endocarditis. Indian J Pathol Microbiol, 2000 Oct, 43(4), 429 - 31 Identification of yeasts from clinical specimens by oxidase test; Kumar S et al.; A total of 100 yeasts and yeast like fungi isolates from clinical specimens were negative for oxidase production on Sabouraud dextrose agar . When grown on Columbia agar, chocolate agar, tryptose agar, Mueller-Hinton agar, brain heart infusion and a medium resembling Sabouraud's dextrose agar but with starch instead of dextrose, all the isolate of Candida albicans (55), C . guilliermondii (6), C . parapsilosis (14), C . tropicalis (6), C . pseudotropicalis (6) and Crytococcus neoformans (2) were positive for oxidase producation . Torulopsis glabrata (2), Saccharomyces cervisiae (2) and two out of seven isolates of C . krusei were negative for oxidase test. Biochim Biophys Acta, 2001 Jan 26, 1517(2), 288 - 92 Isolation and expression of a gene (CGR1) regulated during the yeast-hyphal transition in Candida albicans; Cho T et al.; We used RNA fingerprinting of arbitrarily primed PCR to isolate genes upregulated during the yeast-hyphal transition in Candida albicans . The sequence and expression of one of these genes (CGR1, Candida growth regulation) are presented . Our results suggest that CGR1 expression is associated with a growth cessation of yeast cells, a prerequisite for germination in this organism. Biochim Biophys Acta, 2001 Feb 9, 1545(1-2), 86 - 95 Is salivary histatin 5 a metallopeptide? Gusman H, Lendenmann U, Grogan J, Troxler RF, Oppenheim FG. Histatins are small histidine-rich salivary polypeptides which exhibit antimicrobial activity against Candida albicans . This antimicrobial activity has been ascribed in part to a high content of basic amino acids . However, unlike most other antimicrobial proteins histatins have a high content of histidine, tyrosine and acidic amino acids known to participate in metal ion coordination . This study was conducted to test whether histatin 5 could bind zinc and copper which are metals present in salivary secretions and whole saliva . Physical binding parameters and spectral properties of zinc- and copper-histatin complexes were investigated in order to obtain direct evidence of these interactions . A spectrophotometric competition assay using the metallochromic indicator murexide showed that histatin 5 dissociates metal indicator complexes containing zinc or copper ions . Absorption spectra of histatin 5 at increasing copper chloride concentrations resulted in higher absorbance in the 230-280 nm wavelength range and this spectral change was saturated at a peptide:metal molar ratio of approx . 1:1 . A corresponding band was observed in the visible range of the spectrum with a maximum and molar extinction coefficient corresponding to that of copper binding to an ATCUN motif . Quantitative assessment of zinc and copper binding to histatin 5 using isothermal titration calorimetry revealed at least one high affinity site for each metal, with binding constants of 1.2x10(5) and 2.6x10(7) M(-1), respectively . These results indicate that histatin 5 exhibits metallopeptide-like properties . The precise biological significance of this has not yet been established but histatins may contribute significantly to salivary metal binding capacity. Genome, 2001 Apr, 44(2), 305 - 8 A system of rapid isolation of end-DNA from a small amount of fosmid DNA, with vector-based PCR for chromosome walking; Chibana H et al.; The pBAC 108L and pFos 1 vectors were developed as stable propagation vectors which, due to their extremely low copy number, facilitate the cloning of a large-sized insert containing repeated DNA . However, the low copy number requires laborious end-DNA preparation for end sequencing and chromosome walking . Here we describe efficient methods for end-DNA isolation . The entire process, including small-scale DNA preparation, restriction digestion, self-ligation, and PCR with vector-based primers, is carried out in 96-well formats . Using a Fosmid library of genomic DNA of Candida albicans, PCR products ranging in size from 0.1 to 8 kbp were generated from 118 end sequences in 140 reactions from 70 Fosmid clones . A single or a prominent band was found in 101 of these reactions . Twenty-six of these bands were tested for walking and all of them proved to be specific . Thus, the system overcomes the disadvantage caused by low copy number . This system allows rapid physical mapping of genomes, and is adaptable for several other vectors including BAC (bacterial artificial chromosome), PAC (P1-derived artificial chromosome), and YAC (yeast artificial chromosome). J Nucl Med, 2001 May, 42(5), 788 - 94 99mTc-labeled antimicrobial peptides for detection of bacterial and Candida albicans infections; Welling MM et al.; This study compared the possibilities and limitations of 99mTc-labeled synthetic peptides derived from two human antimicrobial peptides, namely, ubiquicidin (UBI) and lactoferrin (hLF), for the scintigraphic detection of bacterial and fungal infections in mice and rabbits . The rationale of our approach was that selected peptides accumulate in infected areas but not in sterile inflammatory lesions, because they bind preferentially to microorganisms . 99mTc-labeled human neutrophil peptides (defensins), ciprofloxacin, and human polyclonal IgG were included as control agents . METHODS: 99mTc-labeled peptides and control agents were injected intravenously into animals that had been injected intramuscularly 18 h earlier with multidrug-resistant Staphylococcus aureus, Klebsiella pneumoniae, or fluconazole-resistant Candida albicans . Sterile inflammatory sites were induced by the injection of heat-killed microorganisms or lipopolysaccharide (LPS) into the thigh muscle . Up to 4 h after injection, the accumulation of 99mTc-labeled compounds in the infected/inflamed thigh muscles was determined using scintigraphic techniques and radioactivity counts in dissected tissues . RESULTS: Scintigraphy revealed that 99mTc-labeled peptides UBI 29-41, UBI 18-35, UBI 31-38, hLF 1-11, and defensins, which showed preferential in vitro binding to microorganisms in a former study, accumulated at a significantly higher rate (P < 0.01) in bacterial and C . albicans infections in mice and rabbits than in inflamed tissues induced by heat-killed microorganisms or by LPS . No significant difference in the accumulation of 99mTc-labeled ciprofloxacin was observed between infected and sterile inflamed thigh muscles in mice . CONCLUSION: 99mTc-labeled antimicrobial peptides UBI 29-41, UBI 18-35, UBI 31-38, hLF 1-11, and defensins accumulate significantly in tissues infected with gram-positive and gram-negative bacteria and C . albicans . Significantly lower (P < 0.01) accumulation of these peptides occurs in sterile inflamed tissues . These data indicate that the peptides preferentially tag microorganisms at the site of infection, which is in agreement with their preferential binding to the microorganisms in vitro and in vivo . 99mTc-labeled ciprofloxacin does not distinguish between infections and sterile inflammatory lesions, which implies that its specificity for the detection of bacterial infections is not warranted. Diagn Microbiol Infect Dis, 2001 Mar, 39(3), 165 - 8 First isolation of Candida dubliniensis in Rio Grande do Sul, Brazil; Alves SH et al.; Candida dubliniensis is a newly recognized species closely phylogenetically related to Candida albicans and is commonly associated with oral candidiasis in human immunodeficiency virus-positive patients . In this paper we report the isolation of three strains of C . dubliniensis, from AIDS patients, in the state of Rio Grande do Sul (Brazil) . The phenotypic identification was based on germ tube emission, abundant production of chlamydospores, assimilation of sucrose but not of xylose and the inability to grow at 42 degrees C . Randomly amplified polymorphic DNA (RAPD) analysis and genomic DNA sequencing confirmed the distinct genetic nature C . dubliniensis . Topics related to the epidemiology, isolation, phenotypical and genotypical identification of C . dubliniensis are also discussed. J Trop Pediatr, 2001 Apr, 47(2), 77 - 80 Endoscopic and histopathological evaluation of preschool children with chronic diarrhoea; Mishra OP et al.; Fifty-seven children with chronic diarrhoea, aged 1-5 years, were studied . Protein-energy malnutrition was present in 49 (85.9 per cent) children . Anaemia (89.5 per cent), presence of mucus and blood in stool (66.6 per cent), abdominal distension (52.6 per cent), and abdominal pain (28.1 per cent) were the common clinical findings at admission . The enteropathogens isolated from the stool of 68.4 per cent of patients were Escherichia coli (19.3 per cent), Candida albicans (12.3 per cent), E . histolytica (8.8 per cent), and Giardia lamblia (7 per cent) . Secondary lactose intolerance was present in 21 per cent of children, endoscopic appearance was abnormal in 23.3 per cent of children and the commonest finding was chronic duodenitis (16.7 per cent) . Abnormal histopathology was observed in 73.3 per cent of cases and villous atrophy with mononuclear cell infiltration (56.7 per cent) was the most common abnormality detected . The mean duration of diarrhoea had no impact on endoscopic appearance but it significantly affected the histopathological changes . However, no correlation was found in endoscopic and histopathological lesions in relation to malnutrition and aetiological agents. Nippon Ishinkin Gakkai Zasshi, 2001, 42(2), 87 - 90 {A case of Folliculitis barbae Candidomycetica}; Hattori A et al.; A 71-year-old man was referred to our department on January 30, 1998 with hard red papules that had developed on the philtrum in mid-January . On January 2, the patient had received high-dose steroid therapy (pulse therapy) for cluster asthma attacks and antibiotics at the Department of Internal Medicine of our hospital . Infiltrative, protruding reddish plaques were observed on the philtrum, which contained a number of small pustules at sites corresponding to hair follicles . There was partial opacity and slight irregularity of the nail plates on the first and second toes of the right foot . Fungal elements were detected from a lesion on the mustache and the nail . Histological examination of the lesion on the philtrum revealed infiltration of inflammatory cells comprising neutrophils, lymphocytes, and macrophages around the hair follicles . Beard hair and nail cultures revealed Candida albicans A, indicating that the patient had candidal sycosis and candidal onychia . He was treated with oral fluconazole (100 mg/day) . The lesion was clinically improved within 50-days . Recently, extensive use of steroids and antibiotics has produced an increase in reports of patients with Folliculitis barbae Candidomycetica . We believe that the present case was also induced by high-dose steroid therapy and antibiotics. J Chemother, 2001 Apr, 13(2), 161 - 6 Disk diffusion method for fluconazole susceptibility testing of Candida albicans strains; Yucesoy M et al.; This study evaluated the usefulness of the disk diffusion method by using different media for the susceptibility testing of fluconazole against Candida albicans strains . The susceptibility of 108 clinical isolates of C . albicans against fluconazole were determined by microdilution and disk diffusion methods by using RPMI 1640 agar and 25 microg disks . 93 of these isolates were also tested by disk diffusion technique on four different media (yeast nitrogen base agar, Sabouraud dextrose agar, Mueller Hinton agar and Mueller Hinton methylene blue agar) . The results of the microdilution method were evaluated visually and optically . The disk diffusion results were determined after 24 and 48 hours of incubation . When the 24-hour zone diameters were compared to the minimal inhibitory concentrations determined visually and optically, the best results were obtained for RPMI 1640 agar and yeast nitrogen base agar . The correlation coefficients were r=-0.34, -0.41 and r=-0.33, -0.32 for the first and second media, respectively . The best values (r=-0.29, -0.39) were obtained for Mueller Hinton methylene blue agar when the 48-hour zone diameters were considered . Agreement between the disk diffusion and microdilution methods was best for RPMI 1640, yeast nitrogen base and Mueller Hinton methylene blue agar after 24 hours of incubation (87-89%, 88-90%, 93-96%, respectively) and for Mueller Hinton methylene blue agar after 48 hours of incubation (89-96%) . Disk diffusion method using RPMI 1640, yeast nitrogen base and Mueller Hinton methylene blue agar appears to be a useful, rapid and reliable screening technique for testing the susceptibility of C . albicans strains to fluconazole. Clin Diagn Lab Immunol, 2001 May, 8(3), 585 - 7 New assay for measuring cell surface hydrophobicities of Candida dubliniensis and Candida albicans; Jabra-Rizk MA et al.; Hydrophobic interactions, based on cell surface hydrophobicity (CSH), are among the many and varied mechanisms of adherence deployed by the pathogenic yeast Candida albicans . Recently it was shown that, unlike C . albicans, C . dubliniensis is a species that exhibits an outer fibrillar layer consistent with constant CSH . Previously, C . dubliniensis grown at 25 or 37 degrees C was shown to coaggregate with the oral anaerobic bacterium Fusobacterium nucleatum . C . albicans, however, demonstrated similar coaggregation only when hydrophobic or grown at 25 degrees C . This observation implied that coaggregation of Candida cells with F . nucleatum is associated with a hydrophobic yeast cell surface . To test this hypothesis, 42 C . albicans and 40 C . dubliniensis clinical isolates, including a C . albicans hydrophobic variant, were grown at 25 and 37 degrees C and tested with the established hydrophobicity microsphere assay, which determines CSH levels based on the number of microspheres attached to the yeast cells . The coaggregation assay was performed in parallel experiments . All C . dubliniensis isolates grown at either temperature, hydrophobic 25 degrees C-grown C . albicans isolates, and the C . albicans hydrophobic variant, unlike the 37 degrees C-hydrophilic C . albicans isolates, exhibited hydrophobic CSH levels with the microsphere assay and simultaneously showed maximum, 4+, coaggregation with F . nucleatum . The parallel results obtained for C . dubliniensis using both assays support the use of the CoAg assay both as a rapid assay to determine CSH and to differentiate between C . dubliniensis and C . albicans. J Clin Pathol, 2001 May, 54(5), 362 - 6 Rapid identification of candida species by TaqMan PCR; Guiver M et al.; AIM: To develop and evaluate a TaqMan(TM) polymerase chain reaction (PCR) for the rapid identification and speciation of candida species . METHODS: Species specific primer and probe sets were designed for the identification of Candida albicans, C . parapsilosis, C . tropicalis, C . krusei, C . kefyr, and C . glabrata from clinical isolates in a 5' exonuclease (TaqMan(TM)) assay . The probes were labelled with three fluorescent dyes to enable differentiation between species when three primer and probe sets were combined in two multiplexes . The specificity of these assays was evaluated against a range of National Collection of Pathogenic Fungi strains, clinical isolates of yeast, bacterial and viral pathogens . RESULTS: The primer and probe sets have been shown to be 100% specific for their respective species; there was no crossreaction between any set and human DNA, or extracts from other candida species, fungal, bacterial, or viral pathogens tested . Extracts from two clinical isolates, originally identified as C albicans on the basis of germ tube formation, were not amplified by any of the primer and probe sets . These isolates have been putatively re-identified as C dubliniensis after sequencing of the variable internal transcribed spacer region ITS2 and lack of growth at 45 degrees C . CONCLUSION: This TaqMan assay provides a rapid alternative to conventional culture based techniques for the identification and speciation of the most frequently isolated candida species . The simple extraction method followed by TaqMan PCR can identify the six species mentioned in four hours. J Antimicrob Chemother, 2001 May, 47(5), 527 - 36 Mechanisms of fluconazole resistance in Candida albicans isolates from Japanese AIDS patients; Maebashi K et al.; Four Candida albicans isolates, TIMM 3163, TIMM 3164, TIMM 3165 and TIMM 3166, with reduced fluconazole susceptibility were obtained from three AIDS patients in Japan, and the mechanisms of their drug resistance were studied . All isolates showed lower levels of intracellular accumulation of fluconazole than ATCC 10231, a susceptible control strain of C . albicans . Increased amounts of CDR1 and CDR2 mRNA encoding putative ATP binding cassette (ABC) transporters were associated with the azole resistance of all TIMM isolates, apart from TIMM 3164 . In addition, increased Cdr1p levels were immunodetected in the cell membrane fractions of all the TIMM strains except for TIMM 3164 . Gene amplification was not responsible for CDR1 overexpression and there were no significant differences in the mRNA levels of CDR3 or CDR4 (ABC transporters) in the azole-susceptible and -resistant cells . CaMDR1 (a major facilitator superfamily) gene expression was not observed in any of the resistant isolates or the control strain . These results suggest that energy-dependent drug efflux associated with increased expression of CDR1 and CDR2 is involved in the fluconazole resistance mechanisms in two of the four isolates, TIMM 3165 and TIMM 3166 . TIMM 3164 demonstrated energy-dependent drug efflux without overexpression of CDR1-4 or CaMDR1, indicating that some other pump may be operating . Despite showing low levels of drug efflux and overexpression of CDR1 and CDR2, efflux in TIMM 3163 was not energy dependent, suggesting that the expressed Cdr1p non-functional Cdr1p and that other resistance mechanisms may operate in this strain. J Antimicrob Chemother, 2001 May, 47(5), 513 - 9 HWY-289, a novel semi-synthetic protoberberine derivative with multiple target sites in Candida albicans; Park KS et al.; The antifungal properties of 515 synthetic and semi-synthetic protoberberines were investigated . HWY-289 was chosen for further study because it exhibited the most significant anti-Candida activity (MICs were 1.56 mg/L for Candida albicans and Candida krusei; 6.25 mg/L for Candida guilliermondii) but did not demonstrate toxicity in rats . HWY-289 inhibited the incorporation of L-{methyl-(14)C}methionine into the C-24 of ergosterol in whole cells of C . albicans (IC(50) 20 microM) . However, HWY-289 (100 microM) had no effect on mammalian cholesterol biosynthesis in rat microsomes while miconazole (100 microM) was a potent inhibitor of cholesterol biosynthesis under identical assay conditions . A second major target site for HWY-289 was identified that involves cell wall biosynthesis in C . albicans . HWY-289 was a potent inhibitor of the chitin synthase isozymes CaCHS1 and CaCHS2, with IC(50) values of 22 microM for each enzyme . The effect was highly specific in that HWY-289 had no significant effect on C . albicans CaCHS3 (IC(50) > 200 microM) . Thus, HWY-289 compared favourably with well-established antifungal agents as an inhibitor of the growth of Candida species in vitro, and may have considerable potential as a new class of antifungal agent that lacks toxic side effects in the human host. J Clin Microbiol, 2001 May, 39(5), 2015 - 6 Evaluation of a reformulated CHROMagar Candida; Jabra-Rizk MA et al.; CHROMagar Candida is a differential culture medium for the isolation and presumptive identification of clinically important yeasts . Recently the medium was reformulated by Becton Dickinson . This study was designed to evaluate the performance of the new formula of CHROMagar against the original CHROMagar Candida for recovery, growth, and colony color with stock cultures and with direct plating of clinical specimens . A total of 90 stock yeast isolates representing nine yeast species, including Candida dubliniensis, as well as 522 clinical specimens were included in this study . No major differences were noted in growth rate or colony size between the two media for most of the species . However, all 10 Candida albicans isolates evaluated consistently gave a lighter shade of green on the new CHROMagar formulation . In contrast, all 26 C . dubliniensis isolates gave the same typical dark green color on both media . A total of 173 of the 522 clinical specimens were positive for yeast, with eight yeast species recovered . The recovery rates for each species were equivalent on both media, with no consistent species-associated differences in colony size or color . Although both media were comparable in performance, the lighter green colonies of C . albicans isolates on the new CHROMagar made it easier to differentiate between C . albicans and C . dubliniensis isolates . In conclusion, the newly formulated Becton Dickinson CHROMagar Candida medium is as equally suited as a differential medium for the presumptive identification of yeast species and for the detection of multiple yeast species in clinical specimens as the original CHROMagar Candida medium. J Bacteriol, 2001 May, 183(10), 3211 - 23 CAP1, an adenylate cyclase-associated protein gene, regulates bud-hypha transitions, filamentous growth, and cyclic AMP levels and is required for virulence of Candida albicans; Bahn YS et al.; In response to a wide variety of environmental stimuli, the opportunistic fungal pathogen Candida albicans exits the budding cycle, producing germ tubes and hyphae concomitant with expression of virulence genes, such as that encoding hyphal wall protein 1 (HWP1) . Biochemical studies implicate cyclic AMP (cAMP) increases in promoting bud-hypha transitions, but genetic evidence relating genes that control cAMP levels to bud-hypha transitions has not been reported . Adenylate cyclase-associated proteins (CAPs) of nonpathogenic fungi interact with Ras and adenylate cyclase to increase cAMP levels under specific environmental conditions . To initiate studies on the relationship between cAMP signaling and bud-hypha transitions in C . albicans, we identified, cloned, characterized, and disrupted the C . albicans CAP1 gene . C . albicans strains with inactivated CAP1 budded in conditions that led to germ tube formation in isogenic strains with CAP1 . The addition of 10 mM cAMP and dibutyryl cAMP promoted bud-hypha transitions and filamentous growth in the cap1/cap1 mutant in liquid and solid media, respectively, showing clearly that cAMP promotes hypha formation in C . albicans . Increases in cytoplasmic cAMP preceding germ tube emergence in strains having CAP1 were markedly diminished in the budding cap1/cap1 mutant . C . albicans strains with deletions of both alleles of CAP1 were avirulent in a mouse model of systemic candidiasis . The avirulence of a germ tube-deficient cap1/cap1 mutant coupled with the role of Cap1 in regulating cAMP levels shows that the Cap1-mediated cAMP signaling pathway is required for bud-hypha transitions, filamentous growth, and the pathogenesis of candidiasis. J Bacteriol, 2001 May, 183(10), 2971 - 8 Divergence in fitness and evolution of drug resistance in experimental populations of Candida albicans; Cowen LE et al.; The dissemination and persistence of drug-resistant organisms in nature depends on the relative fitness of sensitive and resistant genotypes . While resistant genotypes are expected to be at an advantage compared to less resistant genotypes in the presence of drug, resistance may incur a cost; resistant genotypes may be at a disadvantage in the absence of drug . We measured the fitness of replicate experimental populations of the pathogenic yeast Candida albicans founded from a single progenitor cell in a previous study (L . E . Cowen, D . Sanglard, D . Calabrese, C . Sirjusingh, J . B . Anderson, and L . M . Kohn, J . Bacteriol . 182:1515-1522, 2000) and evolved in the presence, and in the absence, of the antifungal agent fluconazole . Fitness was measured both in the presence and in the absence of fluconazole by placing each evolved population in direct competition with the drug-sensitive ancestor and measuring the reproductive output of each competitor in the mixture . Populations evolved in the presence of drug diverged in fitness . Any significant cost of resistance, indicated by reduced fitness in the absence of drug, was eliminated with further evolution . Populations evolved in the absence of drug showed more uniform increases in fitness under both conditions . Fitness in the competition assays was not predicted by measurements of the MICs, doubling times, or stationary-phase cell densities of the competitors in isolation, suggesting the importance of interactions between mixed genotypes in competitions. FEMS Microbiol Lett, 2001 Apr 20, 198(1), 37 - 43 Involvement of aspirin-sensitive oxylipins in vulvovaginal candidiasis; Deva R et al.; 3(R)-Hydroxyoxylipins are produced via an aspirin-sensitive pathway in Candida albicans, an abundant pathogen in vulvovaginal candidiasis . In the present study, we have investigated the effect of aspirin on vaginal isolates of C . albicans from patients with recurrent candidiasis . Aspirin alone and with clotrimazole, a commonly used drug, strongly suppressed growth of C . albicans . 3(R)-Hydroxyoxylipins, which were selectively located in hyphae and other filamentous structures, but not in free blastospores, were almost totally suppressed by aspirin . Moreover, C . albicans stimulated prostaglandin E(2) (PGE(2)) production in HeLa cells . PGE(2) is a stimulus for germ tube formation in C . albicans . We conclude therefore that the administration of aspirin should be beneficial in the treatment of vulvovaginal candidiasis by dual ways: (i) by inhibition of 3(R)-hydroxyoxylipin formation, and (ii) by inhibition of PGE(2) formation in the infected host tissue. ASDC J Dent Child, 2001 Jan-Feb, 68(1), 33 - 6, 10 Relation of oral yeast infection in Brazilian infants and use of a pacifier; Mattos-Graner RO et al.; The frequency of oral yeast ingestion and its relationship with sucking and feeding habits was described in children from one to 18 months of age . Yeasts were detected in 58.3 percent of children and the most prevalent species were Candida parapsilosis and Candida albicans . The use of a pacifier was positively associated with the frequency of yeast infection and with the levels of these microorganisms in the mouth . No relationship was detected between the prevalence of yeast and breast-feeding or bottle-feeding habits . The results suggest that use of a pacifier is an important local factor in the colonization and proliferation of yeast in the oral cavity. J Prosthodont, 2000 Dec, 9(4), 184 - 8 Candida albicans colonization of surface-sealed interim soft liners; Olan-Rodriguez L et al.; PURPOSE: This in-vivo investigation evaluated the effect of 2 denture sealer agents on the microbial colonization of a newly placed soft interim denture liner during a period of 14 days . MATERIALS AND METHODS: An interim soft denture liner (Coe-Soft; GC America, Alsip, IL) was coated with 2 different denture surface sealants (Palaseal {Heraeus Kulzer, Irvine, CA} and Mono-Poly {Plastodent, New York, NY}) . Three rectangular wells of 1 cm wide x 2 cm long x 2 mm deep were placed in the intaglio of 10 maxillary complete dentures and filled with the soft liner material . The soft liner surface was treated with Palaseal (first well) and Mono-Poly (second well), and the unsealed (third well) was used as a control . These were exposed to the oral cavity for 14 days . The effect the sealant had in the prevention of Candidal colonization in vivo of the soft liner material was evaluated . Microbiological specimens were recovered from all samples and cultivated . Microbiological data from the control and 2-test samples in each denture were tabulated, and statistical analyses were performed . RESULTS: This investigation showed clear differences (p <.001) between the sealed and unsealed soft liners . The sealed material showed significantly less colonization by yeast and bacteria . Intercomparison of the surface denture sealers, Palaseal versus Mono-Poly, showed no statistically significant differences (p < .005) in total yeast or bacterial colonization . CONCLUSION: Coating of Coe-Soft denture liner with either Palaseal or Mono-Poly significantly decreased yeast and bacterial colonization . . J Prosthet Dent, 2001 Apr, 85(4), 352 - 6 Effects of triclosan on the cytotoxicity and fungal growth on a soft denture liner; Lefebvre CA et al.; STATEMENT OF PROBLEM: Contamination of removable prostheses with microorganisms, particularly Candida albicans, is a common clinical problem . Microban, a broad-spectrum antimicrobial containing triclosan, recently has been proposed to inhibit microbial growth . PURPOSE: This study aimed to determine whether the addition of Microban to PermaSoft denture liner prevents the growth of C albicans and affects the cytotoxicity of the PermaSoft material . MATERIAL AND METHODS: Experimental specimen disks (5 x 1 mm each) with and without incorporated Microban were fabricated aseptically (n = 6) against polyester film to produce a smooth surface . To assess the cytotoxic effect of Microban, the MTT assay was used . To determine the effect of Microban on the growth of C albicans, disks were placed in Transwell dishes, covered with Sabouraud's broth containing an ATCC strain of C albicans, and incubated at 37 degrees C for 24 hours . Wells containing fluorocarbon resin disks or broth alone served as controls . The disks were rinsed to remove unattached C albicans and then sonicated in sterile water to remove surface organisms . Serial dilutions of the water extracts were plated on Sabouraud's agar and returned to the incubator for 24 hours . Colonies were counted with a Brunswick Colony Counter . Growth of C albicans in the internal aspects of the specimens was determined in a manner as previously described, with the exception that the specimens were sonicated to remove surface organisms, minced, and sonicated once more before making serial dilutions . The results were compared with ANOVA and Tukey intervals (alpha=.05) . RESULTS: The number of colonies formed ranged from 17 to 31 x 10(5) (mean = 23 +/- 4 x 10(5)) and 14 to 69 x 10(5) (mean = 32 +/- 20 x 10(5)) for the PermaSoft with and without Microban groups, respectively . There was no statistically significant difference between PermaSoft with and without Microban . CONCLUSION: The addition of Microban did not significantly alter the cytotoxicity of the PermaSoft denture lining material or reduce the adherence of viable C albicans to the surface of PermaSoft material after 24 hours. Gastroenterol Clin Biol, 2001 Feb, 25(2), 161 - 3 {Is endoscopic diagnosis of Candida albicans esophagitis reliable? Correlations with pathology and mycology}; Redah D et al.; AIM OF STUDY: To assess the reliability of endoscopic diagnosis of Candida albicans esophagitis.PATIENTS AND METHODS: A case - control prospective study was carried out from November 1997 to July 1998 at the Campus Teaching Hospital of Lome, in patients with esophagitis macroscopically suggestive of Candida albicans origin at upper digestive endoscopy . Fifteen subjects with normal endoscopy served as controls . Esophageal biopsies for mycologic and pathological examination were performed, as well as HIV serology.RESULTS: During the study period, 26 of the 850 endoscopies performed in our Unit revealed an esophagitis suggestive of Candida albicans origin . Mycology confirmed the presence of filamentous form of Candida albicans in 23 patients and pathology showed non-specific lesions of esophagitis, 20 with intramucous hyphae . HIV serology was positive in 19/23 patients (82.6%) and in 1/15 controls (6.6%) . Sensitivity and specificity of upper GI endoscopy for the diagnosis of Candida albicans were 100 and 83.3% respectively; positive and negative predictive values were 88.5 and 100%, respectively.CONCLUSION: Upper digestive endoscopy is a reliable method for the diagnosis of Candida albicans esophagitis . However, mycological confirmation is warranted. Eur J Med Res, 2001 Feb 28, 6(2), 71 - 4 Interstitial lung disease induced by endogenous Candida albicans; Schreiber J et al.; We report on a 64-year old woman with an interstitial lung disease which had characteristics of hypersensitivity pneumonitis . Severe febrile attacks with impairment of ventilation and diffuse poorly defined radiodensities and ground glass opacities on chest x-ray occured repeatedly . Laboratory data showed hypoxemia, leukopenia and circulating Candida albicans (C.a.)-antigen . Bronchoalveolar lavage revealed an increase in neutrophils . Transbronchial biopsies showed lymphocytic alveolitis, bronchiolitis obliterans and epitheloid cell granulomas . IgG and IgA and the lymphocyte proliferation assay were positive with C.a.-antigen . C.a . was detected in the feces . Intradermal skin test with C.a . showed a positive immediate and late phase reaction and inhalative provocation test with C.a.-antigen was positive . After antimycotic treatment the symptoms resolved completely and long-lasting . We conclude that the disease was induced by C.a.-antigen reaching the lungs from the intestinal tract via the bloodstream. Eur J Med Chem, 2001 Feb, 36(2), 147 - 63 A feature based pharmacophore for Candida albicans MyristoylCoA: protein N-myristoyltransferase inhibitors; Karki RG et al.; A three-dimensional pharmacophore model has been generated for Candida albicans MyristoylCoA: protein N-myristoyltransferase (NMT) inhibitors, using the software program CATALYST . The in vitro NMT inhibitory activity of a series of peptidic inhibitors was used for pharmacophore generation . The effect of altering the control parameters and feature selection was studied to arrive at the pharmacophore model . The selection of the best hypothesis model was based on the total cost, predictive ability, difference in the cost from the null hypothesis and alignment of the training set compounds on to the hypothesis . The pharmacophore model selected has four features; one hydrophobic, two hydrogen bond acceptor and one positive ionisable function . Groups identified as necessary by scanning alanine mutagenesis studies of the peptidic substrate of C . albicans NMT, have been identified as pharmacophore features . Comparison of the ligand binding with the enzyme in the crystal structure of NMT and that proposed by the phamacophore is consistent . The pharmacophore thus generated can be used as a template for designing non-peptidic inhibitors of NMT. Int J Med Microbiol, 2001 Mar, 290(8), 659 - 68 Suppression of type 2 NO-synthase activity in macrophages by Candida albicans; Schroppel K et al.; Macrophages (Mphi) are important for the defence against experimental disseminated candidiasis . Nitric oxide (NO) generated by the inducible isoform of NO-synthase (iNOS or NOS2) is thought to contribute to candidacidal effector functions by activated Mphi . In vitro, however, Mphi cannot control the growth and hyphal formation of Candida (C.) albicans . Using mouse peritoneal exudate Mphi stimulated with IFN-gamma and LPS, we examined the effect of C . albicans on NO synthesis, NOS2 enzyme activity and macrophage survival . C . albicans effectively inhibited the production of NO via suppression of total NOS2 protein and enzyme activity . Hyphal formation of C . albicans and direct interaction with host cells was required for maximum inhibition of NO production, whereas non-filamentous C . albicans mutants released soluble products that effected only partial inhibition . Ultimately, Mphi underwent apoptotic cell death after infection with C . albicans wild-type strains capable of hyphal formation, indicated by loss of the mitochondrial membrane potential and onset of chromatin degradation . NO suppression and Mphi killing are potent activities of C . albicans that may augment virulence of C . albicans. Chemotherapy, 2001 May-Jun, 47(3), 215 - 8 Prospective evaluation of the impact of amoxicillin, clarithromycin and their combination on human gastrointestinal colonization by Candida species; Maraki S et al.; BACKGROUND: Amoxicillin and clarithromycin have been used extensively for the eradication of Helicobacter pylori . However, no study has examined the impact of their combination on the Candida albicans concentration of the gastrointestinal (GI) tract . This is the first study examining and comparing directly the effect of amoxicillin, clarithromycin and their combination on the C . albicans concentration of the human GI tract . METHODS: Thirty-three adult patients (11 in each antibiotic group) were studied prospectively . Quantitative stool cultures for Candida were conducted at the beginning, the end and 1 week after the discontinuation of antibiotic treatment . RESULTS: All three regimens increased the GI colonization in patients by Candida . The combination of amoxicillin with clarithromycin caused the highest increase; however, this was not statistically significant . CONCLUSION: Amoxicillin and clarithromycin used either alone or in combination cause a small to moderate increase in GI colonization by Candida . Hence, these drugs could be safely used in patients at risk for candidiasis originating from the GI tract . Vet Res Commun, 2000 Dec, 24(8), 505 - 15 Evaluation of oxygen-dependent immunodefences of the polymorphonuclear cells of some tropical ruminants; Sahoo G et al.; Activation of polymorphonuclear cells (PMNs) leads to the formation of superoxide, which is in turn dismutated to H2O2 by superoxide dismutase (SOD) and is partly responsible for oxygen-dependent microbicidal activity . However, no comparative information is available on the effect of SOD inhibition before PMN activation to allow simulation of the SOD defects that are known to occur in some ruminants . This paper attempts to examine the degranulative and phagocytic responses in buffalo, cattle and goat PMNs exposed to diethyldithiocarbamate, a known SOD inhibitor . The activity of glutathione peroxidase and reductase was increased in the presence of SOD inhibitor . On activation, H2O2 production increased significantly (p < 0.01), while SOD inhibition before the activation of PMNs caused a significant decline in the production of H2O2 (p < 0.05) in all the species studied . There was a significant increase (p < 0.05) in the phagocytosis of Candida albicans spores by buffalo PMNs activated with opsonized zymosan . Activation of bovine PMNs after exposure to the SOD inhibitor resulted in a significant decline (p < 0.05) in phagocytic activity; in the other species, the two values only approached significance . Among the activators, opsonized zymosan caused a significant increase in phagocytic activity as compared to lipopolysaccharide, particularly in the PMNs of buffaloes (p < 0.05) . Increased fungicidal activity (p < 0.05) occurred with opsonized zymosan-activated PMNs of all the species studied . The fungicidal activity was found to decline in PMNs exposed to SOD inhibitor before activation (p < 0.05) . Interestingly, the phagocytic activity of caprine PMNs was found to be lower than that of PMNs from cattle (p < 0.05). J Agric Food Chem, 2001 Jan, 49(1), 142 - 5 Mosquitocidal, nematicidal, and antifungal compounds from Apium graveolens L . seeds; Momin RA et al.; The methanolic extract of Apium graveolens seeds was investigated for bioactive compounds and resulted in the isolation and characterization of mosquitocidal, nematicidal, and antifungal compounds sedanolide (1), senkyunolide-N (2), and senkyunolide-J (3) . Their structures were determined by 1H and 13C NMR spectral methods . Compounds 1-3 gave 100% mortality at 25, 100, and 100 microg mL(-1), respectively, on the nematode, Panagrellus redivivus . Com