Candida albicans

Genetics . 2005 Jan 16; {Epub ahead of print}
Chromosome Loss Followed by Duplication is the Major Mechanism of Mating Type Locus Homozygosis in Candida albicans; Wu W et al.; Candida albicans, which is diploid, possesses a single mating type (MTL) locus on chromosome 5, which is normally heterozygous (a/alpha) . To mate, C . albicans must undergo MTL-homozygosis to a/a or alpha/alpha . Three possible mechanisms may be used in this process, mitotic recombination, gene conversion or loss of one chromosome 5 homolog, followed by duplication of the retained homolog . To distinguish between these mechanisms, sixteen spontaneous a/a and alpha/alpha derivatives were cloned from four natural a/alpha strains, P37037, P37039, P75063 and P34048, grown on nutrient agar . Eighteen polymorphic (heterozygous) markers were identified on chromosome 5, six to the left and 12 to the right of the MTL locus . These markers were then analyzed in MTL-homozygous derivatives of the four natural a/alpha strains to distinguish between the three mechanisms of homozygosis . An analysis of polymorphisms on chromosomes 1, 2 and R excluded meiosis as a mechanism of MTL-homozygosis . The results demonstrate that while mitotic recombination was the mechanism for homozygosis in one offspring, loss of one chromosome 5 homolog followed by duplication of the retained homolog was the mechanism in the remaining fifteen offspring, indicating that the latter mechanism is the most common in the spontaneous generation of MTL-homozygotes in natural strains of C . albicans in culture.

Am J Ophthalmol, 2005 Jan, 139(1), 135 - 40
Management of endogenous fungal endophthalmitis with voriconazole and caspofungin; Breit SM et al.; PURPOSE: Voriconazole, a new generation triazole, has been shown to achieve therapeutic intraocular levels after oral administration . Caspofungin is the first approved agent from a new class of antifungals, the echinocandins . This series describes experience at two centers using these novel antifungals to treat endogenous fungal endophthalmitis . DESIGN: Retrospective review . METHODS: Treatment of five patients with Candida endophthalmitis are reviewed . Postmortem intraocular voriconazole concentrations on a sixth patient are presented as well . RESULTS: All patients had systemic cultures positive for Candida species . Three patients had prompt resolution of intraocular mycosis with intravenous and oral voriconazole, caspofungin, or both . The fourth patient with bilateral disease responded well to IV voriconazole and caspofungin but had a recurrence when discharged on oral voriconazole and IV caspofungin . This patient had a bowel resection with an ileostomy; therefore, absorption of oral voriconazole may have been inadequate . Bilateral amphotericin B intravitreal injection ultimately treated this patient . The fifth patient received 100 mug/0.1 ml of intravitreal voriconazole (final vitreous concentration approximately 25 mug/ml) followed by oral voriconazole and responded favorably . Our sixth patient had multisystem failure and passed away 1 week after initiating intravenous voriconazole for non-ocular candidemia . Postmortem HPLC analysis of the aqueous and vitreous revealed voriconazole concentrations of 1.52 mug/ml and 1.12 mug/ml, respectively (MIC(90) of Candida albicans is 0.06 mug/ml) . CONCLUSIONS: Voriconazole and caspofungin appear to be powerful weapons to add to the existing armamentarium against fungal endophthalmitis . Further studies are warranted to define precisely the role of these new agents alone or in combination with other antifungals.

J Antimicrob Chemother . 2005 Jan 13; {Epub ahead of print}
Candida krusei fungaemia: antifungal susceptibility and clinical presentation of an uncommon entity during 15 years in a single general hospital; Munoz P et al.; Candida krusei fungaemia is an uncommon entity described in immunocompromised patients previously exposed to azole agents . From 1988 to 2003, 13 episodes of C . krusei fungaemia (2.3% of all fungaemias) were detected in our institution and compared with 39 Candida albicans controls . Susceptibility testing was carried out with the modified microdilution method according to NCCLS recommendations . Underlying conditions were: HIV infection (4), haematological malignancies (4), organ transplantation (2), abdominal surgery (2) and lactose intolerance (1) . Nine patients (69%) were not neutropenic . In comparison with C . albicans, patients with C . krusei infection had more commonly received antifungal agents (54% versus 15%, P=0.006), had a haematological disease (31% versus 3%, P=0.03), or a transplant (15% versus 3%, P=0.08), were on corticosteroids (47% versus 13%, P=0.01) and were neutropenic (31% versus 0%, P < 0.001) . Patients with C . albicans had more surgical interventions (41% versus 15%, P=0.09) and bladder catheters (61% versus 31%, P=0.05) . The most common origin for C . albicans was a catheter (41% versus 0%; P=0.006) whereas for C . krusei the most common origin was unknown (69% versus 20%; P=0.001) . C . krusei presented more commonly with skin lesions in neutropenic patients (23% versus 5%; P=0.05) . Multivariate analysis of these differential characteristics showed that the only factor that independently predicted the presence of C . krusei fungaemia was the administration of antifungal agents before the fungaemia (RR: 6.4; P=0.009; 95%CI 1.6-25.99) . Overall mortality of C . krusei fungaemia was 38% (C . albicans 49%) . Except for voriconazole (MIC90 0.125 mg/L), azoles and 5-flucytosine had poor activity against C . krusei, whereas amphotericin (MIC90 1 mg/L) and LY-303366 (MIC90 0.06 mg/L) showed good activity . C . krusei fungaemia incidence remains low despite widespread use of azoles . It may occur outside the setting of cancer patients with previous antifungal use . The presence of skin lesions should be a warning sign.

Br J Biomed Sci, 2004, 61(4), 171 - 4
Evaluation of the neutralising capacity of bactec medium for some antibiotics; Nzeako BC et al.; Bactec medium 9240 (Becton Dickinson, MD, USA) is a blood culture medium used routinely at Sultan Qaboos University Hospital (SQUH) . The medium is said to contain substances that can neutralise antibiotics and destroy leucocytes in blood samples . In this study, the ability of the medium to neutralise the effect of some antibiotics and to destroy leucocytes is investigated . Vancomycin, amoxicillin, chloramphenicol, penicillin, gentamicin, fungizone and amikacin at various concentrations were added to separate bottles of Bactec medium 9240 . Escherichia coli (NCTC 10418), Pseudomonas aeruginosa (NCTC 10662), Staphylococcus aureus (NCTC 6571) and Candida albicans (ATCC 10231), each at 1 x 10(4) colony-forming units (CFU)/mL were added separately, depending on the type of antibiotic . The blood samples were incubated at 37 degrees C for seven days under manual and automated blood culture systems . Subcultures were made from the manual system and routine laboratory procedures for detection of positive cultures were followed for the automated system . The Bactec medium was found to neutralise all antibiotics up to a concentration of 100 microg/mL by the automated method but showed some variation in results by the manual system . Leucocytes were destroyed within 24 hours.

Allergy, 2005 Feb, 60(2), 238 - 42
Immediate hypersensitivity to Malassezia furfur and Candida albicans mannans in vivo and in vitro; Kosonen J et al.; Background: Elevated and correlative Malassezia furfur (M . furfur) and Candida albicans (C . albicans) mannan-specific IgE have been demonstrated in atopic eczema dermatitis syndrome (AEDS) of the head, neck and shoulder (HNS) region of the skin . The significance of these antibodies in vivo has not been demonstrated . Methods: Sixty-five AEDS patients with HNS distribution were included . Serum total IgE (S-IgE) and yeast antigen-specific (Cetavlon-purified mannan and whole extract antigens of M . furfur and C . albicans) IgE were measured and skin prick tests (SPT) were performed with the yeast antigens . Results: Mannan-specific IgE and SPT were positive in 51 and 48% of patients with M . furfur and in 42 and 22% with C . albicans, respectively . Whole extract-specific IgE and SPT were positive in 85 and 95% of patients with M . furfur and in 91 and 57% with C . albicans, respectively . The highest correlation between specific IgE and SPT was seen with M . furfur mannan (r = 0.60; P < 0.0001) . Both M . furfur mannan-specific IgE (r = 0.76; P < 0.0001) and SPT (r = 0.44; P = 0.0005) correlated with S-IgE . Conclusions: Mannan-induced immediate hypersensitivity in vivo was demonstrated in SPT . The significant correlation between M . furfur mannan-specific IgE and SPT suggests that mannan is an important allergen in yeast hypersensitive AEDS in vivo.

Klin Lab Diagn, 2004 Nov, (11), 11 - 3
{Circulating immune complexes in the diagnosis of allergic reactions of the immune-complex type}; Allergens of the entomopathogenic fungus Beauveria bassiana; BACKGROUND: Beauveria bassiana is an important entomopathogenic fungus currently under development as a bio-control agent for a variety of insect pests . Although reported to be non-toxic to vertebrates, the potential allergenicity of Beauveria species has not been widely studied . METHODS: IgE-reactivity studies were performed using sera from patients displaying mould hypersensitivity by immunoblot and immunoblot inhibition . Skin reactivity to B . bassiana extracts was measured using intradermal skin testing . RESULTS: Immunoblots of fungal extracts with pooled as well as individual sera showed a distribution of IgE reactive proteins present in B . bassiana crude extracts . Proteinase K digestion of extracts resulted in loss of IgE reactive epitopes, whereas EndoH and PNGaseF (glycosidase) treatments resulted in minor changes in IgE reactive banding patterns as determined by Western blots . Immunoblot inhibitions experiments showed complete loss of IgE-binding using self protein, and partial inhibition using extracts from common allergenic fungi including; Alternaria alternata, Aspergillus fumigatus, Cladosporium herbarum, Candida albicans, Epicoccum purpurascens, and Penicillium notatum . Several proteins including a strongly reactive band with an approximate molecular mass of 35 kDa was uninhibited by any of the tested extracts, and may represent B . bassiana specific allergens . Intradermal skin testing confirmed the in vitro results, demonstrating allergenic reactions in a number of individuals, including those who have had occupational exposure to B . bassiana . CONCLUSIONS: Beauveria bassiana possesses numerous IgE reactive proteins, some of which are cross-reactive among antigens from other fungi . A strongly reactive potential B . bassiana specific allergen (35 kDa)was identified . Intradermal skin testing confirmed the allergenic potential of B . bassiana.

Eukaryot Cell, 2005 Jan, 4(1), 156 - 65
Demonstration of Loss of Heterozygosity by Single-Nucleotide Polymorphism Microarray Analysis and Alterations in Strain Morphology in Candida albicans Strains during Infection; Forche A et al.; Candida albicans is a diploid yeast with a predominantly clonal mode of reproduction, and no complete sexual cycle is known . As a commensal organism, it inhabits a variety of niches in humans . It becomes an opportunistic pathogen in immunocompromised patients and can cause both superficial and disseminated infections . It has been demonstrated that genome rearrangement and genetic variation in isolates of C . albicans are quite common . One possible mechanism for generating genome-level variation among individuals of this primarily clonal fungus is mutation and mitotic recombination leading to loss of heterozygosity (LOH) . Taking advantage of a recently published genome-wide single-nucleotide polymorphism (SNP) map (A . Forche, P . T . Magee, B . B . Magee, and G . May, Eukaryot . Cell 3:705-714, 2004), an SNP microarray was developed for 23 SNP loci residing on chromosomes 5, 6, and 7 . It was used to examine 21 strains previously shown to have undergone mitotic recombination at the GAL1 locus on chromosome 1 during infection in mice . In addition, karyotypes and morphological properties of these strains were evaluated . Our results show that during in vivo passaging, LOH events occur at observable frequencies, that such mitotic recombination events occur independently in different loci across the genome, and that changes in karyotypes and alterations of phenotypic characteristics can be observed alone, in combination, or together with LOH.

Eukaryot Cell, 2005 Jan, 4(1), 95 - 102
Cyclin Cln3p Links G1 Progression to Hyphal and Pseudohyphal Development in Candida albicans; Bachewich C et al.; G(1) cyclins coordinate environmental conditions with growth and differentiation in many organisms . In the pathogen Candida albicans, differentiation of hyphae is induced by environmental cues but in a cell cycle-independent manner . Intriguingly, repressing the G(1) cyclin Cln3p under yeast growth conditions caused yeast cells to arrest in G(1), increase in size, and then develop into hyphae and pseudohyphae, which subsequently resumed the cell cycle . Differentiation was dependent on Efg1p, Cph1p, and Ras1p, but absence of Ras1p was also synthetically lethal with repression of CLN3 . In contrast, repressing CLN3 in environment-induced hyphae did not inhibit growth or the cell cycle, suggesting that yeast and hyphal cell cycles may be regulated differently . Therefore, absence of a G(1) cyclin can activate developmental pathways in C . albicans and uncouple differentiation from the normal environmental controls . The data suggest that the G(1) phase of the cell cycle may therefore play a critical role in regulating hyphal and pseudohyphal development in C . albicans.

Eukaryot Cell, 2005 Jan, 4(1), 90 - 4
The G1 Cyclin Cln3 Regulates Morphogenesis in Candida albicans; Lazo BC et al.; In Saccharomyces cerevisiae, the G(1) cyclin Cln3 initiates the Start of a mitotic cell cycle in response to size and nutrient inputs . Loss of Cln3 delays but does not prevent Start, due to the eventual Cln3-independent transcription of CLN1 and CLN2 . When unbudded cells of the human pathogen Candida albicans were depleted of the G(1) cyclin Cln3 they increased in size but did not bud . Thus, unlike S . cerevisiae, Cln3 is essential for budding in C . albicans . However, eventually the large unbudded cells spontaneously produced filamentous forms . The morphology was growth medium dependent; on nutritionally poor medium the polarized outgrowths fulfilled the formal criteria for true hyphae . This state is stable, and continued growth leads to a hyphal mycelium, which invades the agar substratum . Interestingly, it is also required for normal hyphal development, as Cln3-depleted cells develop morphological abnormalities if challenged with hyphal inducing signals such as serum or neutral pH . Taken together, these results show that, in C . albicans, Cln3 has assumed a critical role in coordinating mitotic cell division with differentiation.

Arch Biochem Biophys, 2005 Feb 15, 434(2), 358 - 64
Identification and optimization of an antimicrobial peptide from the ant venom toxin pilosulin; Zelezetsky I et al.; A template based on positional residue frequencies in the N-terminal stretch of natural alpha-helical antimicrobial peptides was used to prepare sequence patterns and to scan the Swiss-Prot Database, using the ScanProsite tool . This search identified a segment in pilosulin 1, a cytotoxic peptide from the venom of the jumper ant Myrmecia pilosula, as a potential novel antimicrobial peptide sequence . This segment, corresponding to the 20 N-terminal residues, was synthesized and its structural properties and biological activities were investigated . It showed a potent and broad spectrum antimicrobial activity including standard and multi-drug resistant gram-positive and gram-negative bacteria and Candida albicans, confirming the validity of the search method . A rational redesign approach resulting in four amino acid substitutions yielded a variant with improved antibacterial and significantly reduced hemolytic activity.

Lipids, 2004 Aug, 39(8), 737 - 46
Disruption of ergosterol biosynthesis, growth, and the morphological transition in Candida albicans by sterol methyltransferase inhibitors containing sulfur at C-25 in the sterol side chain; Kanagasabai R et al.; The sterol substrate analog 25-thialanosterol and its corresponding sulfonium salt were evaluated for their ability to serve as antifungal agents and to inhibit sterol methyltransferase (SMT) activity in Candida albicans . Both compounds inhibited cell proliferation, were fungistatic, interrupted the yeast-like-form to germ-tube-form transition, and resulted in the accumulation of zymosterol and related delta24-sterols concurrent with a decrease in ergosterol, as was expected for the specific inhibition of SMT activity . Feedback on sterol synthesis was evidenced by elevated levels of cellular sterols in treated vs . control cultures . However, neither farnesol nor squalene accumulated in significant amounts in treated cultures, suggesting that carbon flux is channeled from the isoprenoid pathway to the sterol pathway with minor interruption or redirection until blockage at the C-methylation step . Activity assays using solubilized C . albicans SMT confirmed the inhibitors impair SMT action . Kinetic analysis indicated that 25-thialanosterol inhibited SMT with the properties of a time-dependent mechanism-based inactivator Ki of 5 microM and apparent kinact of 0.013 min(-1), whereas the corresponding sulfonium salt was a reversible-type transition state analog exhibiting a Ki of 20 nM . The results are interpreted to imply changes in ergosterol homeostasis as influenced by SMT activity can control growth and the morphological transition in C . albicans, possibly affecting disease development.

Am J Physiol Regul Integr Comp Physiol . 2005 Jan 6; {Epub ahead of print}
Parotid secretory protein (PSP) is an HDL-associated protein with anticandidal activity; Khovidhunkit W et al.; High-density lipoprotein (HDL) is part of innate immunity, protecting against infection and inflammation . Using a proteomic approach, we identified an amino acid sequence in a hamster HDL protein that showed homology to rat and mouse parotid secretory protein (PSP), a salivary protein secreted from the parotid glands . We cloned the cDNA encoding a putative hamster homologue of rat and mouse PSP . Searches for conserved domains of the protein showed that the carboxy terminus of hamster PSP contains a region homologous to the amino termini of a family of HDL-associated proteins, including lipopolysaccharide binding protein (LBP), cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP) . In mice, PSP was also associated with HDL, but was not detected in very low-density lipoprotein, low-density lipoprotein, or lipoprotein-deficient sera . In addition to salivary glands, we found that PSP mRNA was expressed in lung, testis, and ovary . The level of PSP in HDL was increased after endotoxin injection in hamsters, but not in mice . Recombinant PSP inhibits growth of Candida albicans in culture . In summary, our results showed that PSP is a novel anticandidal protein associated with HDL.

Res Microbiol, 2005 Jan-Feb, 156(1), 115 - 8
Toll-like receptor 2 mediates prostaglandin E(2) production in murine peritoneal macrophages and splenocytes in response to Candida albicans; Villamon E et al.; The involvement of Toll-like receptor 2 (TLR2) and TLR4 in triggering signal transduction pathways leading to prostaglandin E(2) (PGE(2)) production in response to Candida albicans has been studied in cells from wild-type, TLR2-/- and TLR4-/- knockout mice . In vitro PGE(2) production by macrophages challenged with zymosan, yeast or hypha cells was strongly inhibited in TLR2-deficient cells, but not in TLR4-/- cells, as compared to macrophages from wild-type mice . PGE(2) production was dependent on de novo cyclooxygenase-2 (Cox2) synthesis, since unchallenged cells failed to produce PGE(2) and specific Cox2 inhibition during challenge totally blocked PGE(2) production . Similar results were obtained following in vitro challenge of splenocytes from mice intravenously infected with the low-virulent C . albicans PCA2 strain . This indicates that TLR2 is the major receptor that mediates PGE(2) production in response to C . albicans, probably by upregulating Cox2 expression.

Shi Yan Sheng Wu Xue Bao, 2004 Oct, 37(5), 418 - 22
{Study on experimental induction of fluconazole resistance in Candida albicans}; Zhu YN et al.; To investigate the phenotype and genotype variation between the Fluconazole resistant C . albicans isolates and the corresponding susceptible ones, our research established a resistance-induction mode in vitro . Comparisons were done on drug resistance maintainability, metabolic profile and the doubling time in the logarithmic growth phase . Genotypes were determined by ERIC-PCR . The Fluconazole resistant isolates appeared in strain 435, A06, B07 and C01 from total 22 clinical Fluconazole susceptible isolates after being incubated for 45-80 days in YEPD broth with increasing Fluconazole concentration . The parent isolates had a same metabolic profile and a similar growth doubling time to their filial generation . The same ERIC-PCR profiles were also found between the susceptible parents and their resistant filial isolates . The resistant isolates maintained drug resistance for 24 days after growing on drug-free medium . It was supposed that candida albicans had a latent capacity to evolve resistance to azoles under a certain antifungal drug selective pressure, and the acquired resistance could maintain in drug-free media for a certain period . The resistant isolate with no adaptive cost may be prone to vogue among people . ERIC-PCR could be used in epidemiological study as a stable marker.

Klin Oczna, 2004, 106(3 Suppl), 434 - 5
{In vitro studies on antimicrobial properties of silicon oil}; Mackiewicz J et al.; PURPOSE: The aim of the study was to evaluate antimicrobial properties of silicon oil in vitro against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, which are considered the major causative agents of endophthalmitis . MATERIAL AND METHODS: The clinical isolates of representative microorganisms (S . aureus, P . aeruginosa, C . albicans) were selected . The bacteria and the fungus were separately inoculated in PDMS 5000 (produced by AcriMed, Germany) . Control inoculations in physiological saline and sugar bouillon were performed . The samples of 0.01 ml from each medium were diluted, according to serial dilution procedure and inoculate on Petri plate dishes 5% sheep blood agar for bacteria and Sabouraud medium for Candida albicans . After 24 h incubations for bacteria and 48 h incubations for fungus, CFUs were counted . RESULTS: All the microorganisms revealed an apparent decrease in CFUs in PDMS 5000 . The total elimination was observed for S . aureus after 5 days . For P . aerugisosa solitary colonies (less than 25 CFUs) were observed up to 7 days . After 7 days of incubation no growth of P . aeruginosa was observed . High C . albicans CFU values were counted up to 3 day of the incubation . After 5 days single fungal colonies were observed . CFUs of the examined microorganisms declined slightly in physiologic saline . A growth pattern similar to the growth curve of microorganisms was observed in sugar boullion . CONCLUSIONS: Our study indicates that silicon oil could have an antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, which are considered the major causative agents of postoperative endophthalmitis.

Epidemiol Infect, 2004 Dec, 132(6), 1175 - 80
A one-year survey of candidemia in Belgium in 2002; Swinne D et al.; A total of 211 episodes of bloodstream yeast infections in 207 patients, hospitalized in 28 Belgian hospitals participating in a National Surveillance Program, were evaluated . A total of 81% of the patients were more than 50 years of age . Candida albicans was the cause of infections in 55% of patients, 22% were due to C . glabrata and 13% to C . parapsilosis . The most common predisposing factors were antibacterial therapy (42%), residence in an intensive care unit (32.9%) and presence of an intravascular catheter (29.7%) . Most patients had more than one predisposing factor . Fluconazole alone or in association with another antifungal agent was the treatment of choice for 89.7% of the cases . In vitro susceptibility testing of the isolates revealed that 99% were susceptible to amphotericin B, 95% to 5-fluorocytosine, 82% to fluconazole and 69% to itraconazole . Resistance to azoles was more common among C . glabrata isolates in the elderly . We conclude that the frequency of C . albicans infection is decreasing in Belgium and this is associated with the emergence of other species, most notably, C . glabrata.

Yeast, 2005 Jan 15, 22(1), 57 - 70
5-Fluoro-orotic acid induces chromosome alterations in Candida albicans; Wellington M et al.; Treatment of a prototrophic laboratory strain of Candida albicans with 5-fluoro-orotic acid (5-FOA) produced two major types of mutants with chromosomal alterations, 5-FOA-resistant (Foa(R)) and those remaining sensitive (Foa(S)) . Both major types remained Ura(+) . Foa(R) mutants, produced after a long exposure, contained either a duplication of chromosome 4b or an inner enlargement of chromosome 5b . The average mutant frequency was approximately 1.0 x 10(-5) . The reverse mutation of Foa(R) to Foa(S) also caused the loss of either the extra chromosome 4b or the enlarged chromosome 5b, revealing a causal relationship between the resistance and the specific chromosome constitution . The cells remained sensitive after a relatively short 24 h exposure to 5-FOA medium, but the treatment induced non-specific changes in lengths of various chromosomes . Furthermore, Foa(R) type mutants acquired a notable chromosomal and phenotypic instability . Our results indicate the necessity of electrokaryotyping of strains that have been exposed to 5-FOA, especially with studies of gene function and with DNA microarray assays . Copyright (c) 2004 John Wiley & Sons, Ltd.

J Clin Microbiol, 2005 Jan, 43(1), 387 - 92
Purification of Enterocytozoon bieneusi from stools and production of specific antibodies; Sheoran AS et al.; Enterocytozoon bieneusi is clinically the most significant of the microsporidia in humans, causing chronic diarrhea wasting and cholangitis in individuals with human immunodeficiency virus infection and AIDS . Little progress on this infection has been made because of the inability to propagate E . bieneusi in vitro and in vivo, which limits the source of parasite spores to the stools of infected human patients . Given the size and shape of the E . bieneusi spores (1.1 to 1.6 by 0.7 to 1.0 microm) and the lack of specific immune reagents, the identification and purification of large quantities of spores from feces are technically challenging . Consequently, diagnosis relies entirely on PCR, a labor-intensive approach that requires highly skilled personnel . We describe a method for the purification of E . bieneusi spores from human stools and the production of rabbit-specific antisera . Spores were purified by a combination of isopycnic Percoll gradient centrifugation and continuous sucrose gradient centrifugation . Specific polyclonal antibodies raised in mice and rabbits reacted by indirect immunofluorescence with E . bieneusi but not with Encephalitozoon spp., Candida albicans, Staphylococcus aureus, Escherichia coli, or other forms present in human stools.

Microbiology, 2005 Jan, 151(Pt 1), 81 - 9
Generation and functional in vivo characterization of a lipid kinase defective phosphatidylinositol 3-kinase Vps34p of Candida albicans; Gunther J et al.; The phosphatidylinositol (PI) 3-kinase Vps34p of Candida albicans has lipid kinase and autophosphorylation activity and is involved in virulence and vesicular protein transport . In order to characterize the roles of lipid kinase activity, a chimeric Vps34 protein was created which lacks lipid kinase but retains autophosphorylation activity . To this end, six amino acids within the putative lipid-binding site of Vps34p were replaced by the homologous region of the PI 3-kinase-like C . albicans Tor protein . The resulting chimeric Vps34T protein was recombinantly expressed in Escherichia coli and shown to lack lipid kinase activity . The corresponding chimeric VPS34TOR gene was inserted into the genome of C . albicans, and this lipid-kinase-defective strain had a distinctive phenotype compared to those of the wild-type strain SC5314 and the vps34 null mutant . The lipid-kinase-defective strain was non-virulent, and showed altered hyphal growth, reduced adherence, as well as defective vacuole morphology and endosomal vesicle transport . These results demonstrate an important role for the lipid kinase activity of Vps34p in virulence and vesicular protein transport . On the other hand, the lipid-kinase-defective strain and the vps34 null mutant differ in their temperature- and osmotic-stress response . This indicates a possible role for activities different from the lipid kinase function of Vps34p.

J Biol Chem . 2005 Jan 4; {Epub ahead of print}
N-cadherin mediates endocytosis of candida albicans by endothelial cells; Phan QT et al.; Candida albicans is the most common cause of fungal bloodstream infections . To invade the deep tissues, blood-borne organisms must cross the endothelial cell lining of the vasculature . We have found previously that C . albicans hyphae, but not blastospores, invade endothelial cells in vitro by inducing their own endocytosis . Therefore, we set out to identify the endothelial cell receptor that mediates the endocytosis of C . albicans . We determined that endocytosis of C . albicans was not mediated by bridging molecules in the serum, and that it was partially dependent on the presence of extracellular calcium . Using an affinity purification procedure, we discovered that endothelial cell N-cadherin bound to C . albicans hyphae, but not blastospores . N-cadherin also co-localized with C . albicans hyphae that were being endocytosed by endothelial cells . Chinese hamster ovary (CHO) cells expressing human N-cadherin endocytosed significantly more C . albicans hyphae than did CHO cells expressing either human VE-cadherin or no human cadherins . Expression of N-cadherin by the CHO cells resulted in enhanced endocytosis of hyphae, but not blastospores, indicating the selectivity of the N-cadherin mediated endocytosis . Down-regulation of endothelial cell N-cadherin expression with siRNA significantly inhibited the endocytosis of C . albicans hyphae . Therefore, a novel function of N-cadherin is that it serves as an endothelial cell receptor which mediates the endocytosis of C . albicans.

Pharmacotherapy, 2004 Oct, 24(10), 1408 - 11
Caspofungin: a potential cause of reversible severe thrombocytopenia; Lynch J et al.; Caspofungin is an echinocandin agent approved for the treatment of invasive candidiasis and refractory aspergillosis . Compared with amphotericin B, caspofungin has an improved safety profile, but clinical experience with this agent is still accumulating . A 68-year-old man developed reversible severe thrombocytopenia, possibly due to caspofungin, after being successfully treated for Candida albicans endocarditis . Given the limited clinical experience with caspofungin, continued vigilance for unusual and serious adverse events associated with the drug is imperative.

Nucleosides Nucleotides Nucleic Acids, 2004, 23(12), 1889 - 910
Synthesis of some novel 6-benzyl(or substituted benzyl)-2-beta-D-glucopyranosyl-1,2,4-triazolo{4,3-b}{1,2,4}triazines as potential antimicrobial chemotherapeutics; Khalil NS et al.; Glucosidation of the new 8-amino-6-benzyl(or substituted benzyl)-2,8-dihydro-1,2,4-triazolo{4,3-b}{1,2,4}triazin-7(3H)-ones (3a-d) with 2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide 4 gave the corresponding N-glucosides 5a-d . Chemical transformations leading to new functionalities have also been achieved to give compounds 7-12 . Antimicrobial activity of compounds 5a-c against Aspergillus fumigatus, Penicillium italicum, Syncephalastrum racemosum, Candida albicans, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coli is described.

Med Oral Patol Oral Cir Bucal, 2005 Jan-Feb, 10(1), 25 - 31
Isolation of Candida dubliniensis in a teenager with denture stomatitis; Mosca CO et al.; Objectives: Test several methods that allow the differentiation between Candida albicans and Candida dubliniensis, in an attempt to assess whether C . dubliniensis can be recovered from the oral cavity of teenagers wearing orthopedic oral prostheses . Material and Methods: Twelve Candida strains were isolated from the prosthesis as well as the palatal mucosa in contact with the dental prosthesis from 12 teenager patients wearing orthopedic oral prostheses . Differentiation between C . albicans and C . dubliniensis was achieved by a number of phenotypic tests (carbon assimilation by the commercially available ID 32C test, growth at 45 grades C on Sabouraud glucose agar, abundant chlamydospore production on Casein agar, and reactivity with a C . dubliniensis antiserum) and the polymerase chain reaction (PCR) . Serotyping of C . albicans was performed with monoclonal antibody B9E . Results: All 12 patients studied presented a Newton s type 2 denture stomatitis and in every patient the same Candida species were isolated from the prosthesis and the palatal mucosa in contact with the dental prosthesis . CHROMagar Candida and the germ tube test allowed the differentiation of isolates giving green colonies and a positive germ tube test from those giving violet colonies and a negative germ tube test . Only the isolate from patient 8 was stained by the C . dubliniensis antiserum and showed abundant chlamydospore production on Casein agar . Eight isolates did not grow at 45 grades C . Identification of all isolates was obtained by the ID 32C test . C . albicans was identified in 75% of patients, C . glabrata in 16,6% and C . dubliniensis in 8,3% . By using specific primers for typing C . dubliniensis, PCR allowed the identification of patient s 8 isolate as C . dubliniensis genotype 1 . Conclusion: C . dubliniensis can be isolated from the oral cavity of teenagers wearing orthopedic oral prostheses and it is possible and technically amenable, the differentiation between C . albicans y C . dubliniensis using the ID 32C test, the observation of abundant chlamydospore production on Casein agar, the reactivity with a C . dubliniensis antiserum and the PCR.

Jpn J Infect Dis, 2004 Dec, 57(6), 279 - 84
Isolation and characterization Candida spp . In jordanian cancer patients: prevalence, pathogenic determinants, and antifungal sensitivity; Al-Abeid HM et al.; The presence of Candida spp . in the oral cavity was evaluated in 95 cancer patients (57 in-patients and 38 out-patients) and in 65 healthcare workers in Amman, Jordan . Candida carriage occurred in 72.6% of cancer patients and 33.8% of healthcare workers, with Candida albicans being the species most commonly recovered, followed by C . glabrata . In-patients were found to harbor Candida spp . at significantly higher levels than out-patients (P = 0.0044) . The number of adhered C . albicans cells and the secretion of extracellular proteinase was significantly higher in the in-patient group than in the out-patient group (P = 0.0016 and 0.00007, respectively); this significant difference was not observed regarding phospholipase secretion . Antifungal sensitivity testing data suggest that isolates were most sensitive to amphotericin B and nystatin, and least sensitive to miconazole and fluconazole, which are commonly used antifungal agents in Jordan.

East Afr Med J, 2004 Aug, 81(8), 398 - 401
Potential pathogens in the lower genital tract at manual vacuum aspiration for incomplete abortion in Korle Bu Teaching Hospital, Ghana; Lassey AT et al.; OBJECTIVES: To determine the carriage rates of potential pathogens in the lower genital tract and factors associated with colonization among women with incomplete abortion . DESIGN: A cross-sectional study . SETTING: The Manual Vacuum Aspiration room of the Korle-Bu Teaching Hospital, Accra, Ghana . SUBJECTS: Two hundred women undergoing Manual Vacuum Aspiration at the Korle-Bu Teaching Hospital . METHODS: Eligible patients were screened for the presence of organisms in the lower genital tract by microscopy and culture of high vaginal and endocervical swabs . RESULTS: Nearly two-thirds of the patients (64.2%) had potential pathogens in the lower genital tract . Bacterial vaginosis alone was present in 47% and a combination of bacterial vaginosis and Candida albicans was present in 17.2% . Residence in an urban slum showed a significant association with the presence of potential pathogens (Odds ratio 2.6; p-value 0.04) . CONCLUSION: Organisms responsible for bacterial vaginosis were the most frequently isolated potential pathogens in the cervical canal of patients with incomplete abortion at the Korle-Bu Teaching Hospital . Management of these patients should therefore include antibiotic prophylaxis against bacterial vaginosis.

J Drug Target, 2004, 12(7), 425 - 33
Prophylactic role of immunomodulators in treatment of systemic candidiasis in leukopenic mice; Khan MA et al.; In the present study, we have evaluated prophylactic role of various immunomodulators viz . lipopolysachharide, protein A and tuftsin to impart protection against experimental candidiasis in leukopenic mice . Both free as well as liposomised form of nystatin was not effective enough in offering complete cure against less susceptible isolate of Candida albicans (JNMCR) infection in immunodebilitant mice . Interestingly, the pretreatment of leukopenic mice with immunomodulators before challenging them with C . albicans increased therapeutic efficacy of the nystatin against systemic candidiasis . Efficacy of the treatment was evaluated on the basis of survival of the animals as well as fungal load in systemic circulation and various organs viz . liver, kidney, spleen and lungs of the treated animals.

J Asian Nat Prod Res, 2005 Mar, 7(1), 91 - 4
Note; Kang WY et al.; A new triterpene named luculiaoic acid A (1), showing inhibitory activity of a leukaemia cell line, along with eleven known compounds, has been isolated from the ethyl acetate extract of the stems of Luculia pinciana Hook . All the structures were elucidated on the basis of NMR, MS, and IR methods . The activity to inhibit Staphylococcus aureus and Candida albicans of all compounds showed that ursolic acid inhibits the growth of Staphylococcus aureus with an MIC of 0.5 mg ml(-1) and an MBC of 10 mg ml(-1), and scopletin inhibits Candida albicans with an MIC of 1 mg ml(-1) and an MBC of 5 mg ml(-1).

Pesqui Odontol Bras, 2004 Jul-Sep, 18(3), 202 - 7
Oral candidosis by Candida albicans in normal and xerostomic mice; Totti MA et al.; The aim of this study was to analyze the effect of sialoadenectomy on the development of oral candidosis after one or four inoculations of Candida albicans . Initially, a suspension containing 10(8) cells/ml of C . albicans ATCC 36801 was prepared . Seventy-eight sialoadenectomized mice and a similar amount of mice with normal salivary flow received a single inoculation of C . albicans suspension . Another group with a similar number of mice received 4 inoculations . The control group consisted of 6 sialoadenectomized mice and 6 mice with normal salivary flow that were not inoculated with C . albicans . Candidosis development was studied histologically in the tongue of the animals 1, 2, 3, 5, and 8 days after inoculation and at 15-day intervals up to 165 days . According to the results obtained, it could be concluded that sialoadenectomy and a higher frequency of yeast inoculation influenced the presence and extension of candidosis lesions.

Shanghai Kou Qiang Yi Xue, 2004 Dec, 13(6), 544 - 8
{Relationship between the genotype of Candida albicans and oral lichen planus.}; Wu L et al.; PURPOSE: The present study was attempted to probe into the relation between the genotype of Candida albicans and oral lichen planus, analyze the relativity of the genotype between commensal strains and pathogenic strains,to lay a good foundation for the prevention and treatment for candidiasis . METHODS: The present study adopted random amplified polymorphic DNA (RAPD) to analyze 46 Candida species which were isolated from the oral cavity of 9 health adults (N group) and the patients suffering from oral mucosal disease (including 16 lichen planus,LP group and 21 oral candidiasis, OC group) . RESULTS:40 strains were identified as Candida albicans.Different candida species emerged with different genetic types.The genotype of the OC group and N group were similar, genetic homology exits between OC group and N group . There was no distinct similarity in RAPD fingerprinting map of the LP group, which also had significant difference to the genotype of the N group(P<0.001).There was significant difference between OC group and LP group(P<0.001) . CONCLUSION: (1) The RAPD genotype not only can discriminate candida species, but also can distinguish further different types in the same species.(2) Oral candidiasis may originate from commensal strains . Candida albicans isolated from oral candidiasis were endogenous.(3) According to the results of genotypes, there might be no genetic homology between commensal strains and pathogenic strains in LP group.(4) The genotypes of pathogenic strains were not identical in different kinds of oral mucosa diseases.

Eur J Clin Microbiol Infect Dis . 2004 Dec 24; {Epub ahead of print}
Epidemiology and antifungal susceptibility of Candida species isolated from blood: results of a 2-year multicentre study in Spain; Peman J et al.; This study, included in the prospective survey of candidaemia in Europe supported by the European Confederation of Medical Mycology, presents the epidemiological and antifungal susceptibility results of 290 cases of candidaemia (80 in children <15 years old) reported from September 1997 to August 1999 by 19 Spanish hospitals . Presence of an intravenous catheter and previous antibiotic therapy were the most frequent risk factors . The percentages of the four most common species isolated (adults/children) were as follows: Candida albicans (46/36.2), C . parapsilosis (21.9/50), C . tropicalis (12.8/3.75), and C . glabrata (10.1/5) . As initial therapy, fluconazole was preferred in adults (54%) and liposomal amphotericin B in children (58%) . The 30-day mortality rate was 40.6%, and the species most frequently associated with a fatal outcome was C . krusei (60%) . The rates of susceptibility to antifungal agents were as follows: amphotericin B, 91%; flucytosine, 99%; fluconazole, 93.6%; itraconazole, 87.4%; and voriconazole, 92% . These results provide baseline data for future epidemiological and susceptibility studies and for evaluating the impact of new antifungal agents on the distribution of species and the mortality rates associated with candidaemia in Spain.

Infect Immun, 2005 Jan, 73(1), 622 - 6
Intercellular adhesion molecule 1-dependent activation of interleukin 8 expression in Candida albicans-infected human gingival epithelial cells; Egusa H et al.; Increased induction of interleukin 8 (IL-8) and intercellular adhesion molecule 1 (ICAM-1) by oral epithelial cells may play a role in the host defense mechanism in oropharyngeal candidiasis; however, little is known about the expression feature of these molecules on human gingival epithelial cells (HGECs) during Candida albicans infection . In this report we present evidence that neutralization with antibody against ICAM-1 inhibited both the adherence of C . albicans to HGECs and the Candida-induced production of IL-8, suggesting a role for ICAM-1 in recognition and signaling in HGECs to express IL-8 upon infection with C . albicans.

Drug News Perspect, 1998 Apr, 11(3), 185 - 91
Human mycoses and current antifungal therapy; Fromtling RA; The Focus on Fungal Infections meeting has become a popular conference for specialists in medical mycology and antifungal chemotherapy . Highlights of the 8th meeting are reported, with a focus on infection and therapy . Although the incidence of mycoses has increased, the identification of these fungal etiologic agents remains difficult . Mucosal candidiasis caused by endogenous Candida albicans and Candida species remains the most common fungal manifestation in HIV-infected patients, while systemic infection by Aspergillus species also has increased in HIV patients . Two presenters at the meeting debated whether fungal infections in AIDS patients are becoming less common and less important . Various strategies for antifungal therapy in AIDS or HIV-positive patients were presented . Most fungal infections in solid organ transplant patients are due to Candida species or Aspergillus species; however, dematiaceous (dark-pigmented) fungi are becoming more common fungal pathogens in these patients . Antifungal therapy remains difficult in this patient group . The meeting included an overview of the current status of diagnosing fungal infections through serodiagnostic techniques . If properly validated, serology can be useful in fungal diagnosis since antigens and antibodies are easier to detect than the invading organism . Premature infants are at high risk for developing invasive fungal infections . Antifungal drugs have not been tested in controlled clinical trials in these patients, thus therapy is accomplished using adult treatment regimens and anecdotal experience . As regards the new lipid-based formulations of amphotericin B, published clinical studies are only now appearing in the literature and these reports suggest that the new formulations have reduced toxicity and comparable efficacy compared to conventional amphotericin B under various clinical conditions . Correlation of minimum inhibitory concentration (MIC) values with clinical response to therapy is beginning to emerge . Two fungal cell wall-active compounds have recently entered clinical trials: an echinocandin and the chitin-synthesis inhibitor nikkomycin Z . Pradimycin and analogues have been studied through experimental animal models with good success; however, phase I clinical trials suggested drug-related toxicities and development was stopped . New molecular targets also are being investigated in antifungal therapy . (c) 1998 Prous Science . All rights reserved.

Microbiol Immunol, 2004, 48(12), 937 - 43
Characterization of Mechanisms of Fluconazole Resistance in a Candida albicans Isolate from a Japanese Patient with Chronic Mucocutaneous Candidiasis; Kamai Y et al.; We examined the mechanisms of fluconazole resistance in a fluconazole-resistant Candida albicans isolate from a Japanese patient with chronic mucocutaneous candidiasis . It was demonstrated that the highly resistant phenotype of this strain was associated with combined mechanisms of the energy-dependent reduced intracellular accumulation of fluconazole, presumably due to the increased expression of the ATP-binding cassette efflux pump CDR gene(s), and the reduced affinity of the target enzyme, Erg11p, to fluconazole . In particular, the reduced affinity of Erg11p was considered to contribute largely to the fluconazole resistance in the TIMM3209 strain . Biochemical studies indicated that the Erg11p from the TIMM3209 strain showed reduced susceptibility both to fluconazole and itraconazole of cell-free ergosterol biosynthesis, and cytochrome P-450 also showed reduced affinity to fluconazole in the carbon monoxidecytochrome P-450 complex formation assay . We identified two amino acid substitutions, Y132H and G448V, in Erg11p from the TIMM3209 strain . We found that the cytochrome P-450 from the TIMM3209 strain decayed during incubation at 37 C without fluconazole although it is unknown whether or not the phenomenon is linked to the resistant phenotype . These mutations are thought to confer the above-mentioned characteristics to Erg11p.

Nucleic Acids Res, 2005 Jan 1, 33 Database Issue, D358 - 63
The Candida Genome Database (CGD), a community resource for Candida albicans gene and protein information; Arnaud MB et al.; The Candida Genome Database (CGD) is a new database that contains genomic information about the opportunistic fungal pathogen Candida albicans . CGD is a public resource for the research community that is interested in the molecular biology of this fungus . CGD curators are in the process of combing the scientific literature to collect all C.albicans gene names and aliases; to assign gene ontology terms that describe the molecular function, biological process, and subcellular localization of each gene product; to annotate mutant phenotypes; and to summarize the function and biological context of each gene product in free-text description lines . CGD also provides community resources, including a reservation system for gene names and a colleague registry through which Candida researchers can share contact information and research interests . CGD is publicly funded (by NIH grant R01 DE15873-01 from the NIDCR) and is freely available at http://www.candidagenome.org/.

Nucleic Acids Res, 2005 Jan 1, 33 Database Issue, D353 - 7
CandidaDB: a genome database for Candida albicans pathogenomics; d'Enfert C et al.; CandidaDB is a database dedicated to the genome of the most prevalent systemic fungal pathogen of humans, Candida albicans . CandidaDB is based on an annotation of the Stanford Genome Technology Center C.albicans genome sequence data by the European Galar Fungail Consortium . CandidaDB Release 2.0 (June 2004) contains information pertaining to Assembly 19 of the genome of C.albicans strain SC5314 . The current release contains 6244 annotated entries corresponding to 130 tRNA genes and 5917 protein-coding genes . For these, it provides tentative functional assignments along with numerous pre-run analyses that can assist the researcher in the evaluation of gene function for the purpose of specific or large-scale analysis . CandidaDB is based on GenoList, a generic relational data schema and a World Wide Web interface that has been adapted to the handling of eukaryotic genomes . The interface allows users to browse easily through genome data and retrieve information . CandidaDB also provides more elaborate tools, such as pattern searching, that are tightly connected to the overall browsing system . As the C.albicans genome is diploid and still incompletely assembled, CandidaDB provides tools to browse the genome by individual supercontigs and to examine information about allelic sequences obtained from complementary contigs . CandidaDB is accessible at http://genolist.pasteur.fr/CandidaDB.

Zhonghua Er Bi Yan Hou Ke Za Zhi, 2004 Sep, 39(9), 546 - 8
{Immunohistochemical diagnosis of fungus ball in paranasal sinuses}; Li Y et al.; OBJECTIVE: To study the pathogens of fungus balls in paranasal sinuses and establish an immunohistochemical test by which the main opportunistic fungi could be identified . METHODS: Twenty-five patients with fungal balls were treated by surgical removal of the fungus ball in the infected sinuses . The pathogenic fungi on the specimens were identified by means of routine PAS and immunohistochemical staining methods, and the sensitivity of the two methods were compared . RESULTS: The most commonly infected sinus was the maxillary sinus, followed by sphenoid sinus . Pathogens of fungal balls were found to be aspergillus (92%, 23/25) and candida 2 cases (8%) . Aspergillus and candida albicans in infected sinuses could be specifically identified by immunostainning . There was no statistically significant differences of sensitivity between immunostainning and PAS stain . CONCLUSIONS: The main pathogen of the fungus ball was aspergillus . Immunostainning was a rapid and reliable method to identify fungi in infected tissues of paranasal sinuses . It should be widely used in the diagnosis of fungal sinusitis.

J Reprod Med, 2004 Nov, 49(11), 859 - 66
Activated lactoferrin's ability to inhibit Candida growth and block yeast adhesion to the vaginal epithelial monolayer; Naidu AS et al.; OBJECTIVE: To study in vitro growth-inhibitory effects of activated lactoferrin (ALF) against vaginal isolates of Candida species and to measure the ability of ALF to block interactions of Candida albicans and Candida glabrata to the vaginal epithelial (VE) monolayer . STUDY DESIGN: In vitro effects of ALF on growth of C albicans and C glabrata in Sabouraud dextrose (SD) broth were measured as change in broth turbidity by microscale optical density assay . ALF was tested at 5 and 2.5 mg/mL concentrations against 105 yeast cell inoculum at 370 degrees C for 96 hours and compared with native lactoferrin and control (growth in broth without ALF) . VE cells were isolated from human vaginal tissue biopsies to establish a functional monolayer for yeast interaction studies . ALF effects on Candida interactions with the VE monolayer were tested using 3H-thymidine-labeled yeast . Prophylactic (treatment prior to yeast inoculation onto VE) and therapeutic (treatment to detach VE-adherent yeast) potential of ALF (5 mg/mL) was evaluated against vaginal isolates of C albicans strain NTRL809A and C glabrata strain NTRL131G . RESULTS: Growth of Candida species indicated that a 105 yeast inoculum in SD broth proliferated to a stationary growth equilibrium (approximately 10(9) yeast cell density) in 18 hours (approximately 2 hours of generation time) . ALF (5 mg/mL) elicited >96 hours of total stasis (100% growth inhibition) and was significantly effective against both Candida species (p < 0.0001) . At 2.5 mg/mL dilution, ALF sustained total stasis activity to an average of 18 hours and 24 hours for C albicans (n = 5) and C glabrata (n = 5), respectively . Interaction studies indicated avid binding of C albicans (70 - 140 x 10(3) yeast) and C glabrata (50 - 75 x 10(3) yeast) per square centimeter of VE monolayer . ALF-treated VE showed significant blockade (p < 0.05) of yeast adhesion by 33% and 58% with C albicans and C glabrata, respectively . ALF treatment of yeast-VE complexes resulted in significant detachment (p < 0.05) of C albicans and C glabrata, by 58% and 51%, respectively . CONCLUSION: ALF is a natural fungistatic agent with potent yeast adhesion-blocking and detachment properties and is effective against the vaginal pathogens C albicans and C glabrata.

J Radiol, 2004 Nov, 85(11), 1953 - 5
{Candida arthritis of the TM joint complicating chronic otitis media}; Semlali S et al.; Infectious arthritis of the temporomandibular joint is very uncommon, and arthritis of the TM joint as a result of candida albicans infection has not previously been reported . The authors describe a patient treated for chronic otitis media complicated by arthritis of the temporomandibular joint . The diagnosis was made using CT scan and bacteriologic sampling.

Ai Zheng, 2004 Dec, 23(12), 1707 - 9
{Clinical analysis of nosocomial pulmonary fungal infection in patients with cancer.}; Jiang Y et al.; BACKGROUND & OBJECTIVE: Nosocomial fungal infection is an important complication of cancer patients . Among them, pulmonary fungal infection is over 20% . This study was designed to analyze and evaluate risk factors, and clinical features of nosocomial pulmonary fungal infection in patients with cancer . METHODS: In 1 229 patients with malignant tumor, clinical records of 78 patients with nosocomial pulmonary fungal infection were retrospectively analyzed . RESULTS: Pulmonary fungal infection rate was 6.35% (78/1 229) . The major fungus was Candida albicans (68.18%) . The main risk factors were age of >/=50 years (P< 0.005), primary site (lung cancer, P< 0.001), cancer stage (stage IV, P< 0.005), pulmonary radiotherapy (P< 0.001), chemotherapy (P< 0.001), and long-term hospitalization ( >2 weeks,P< 0.005) . CONCLUSION: The effective methods to reduce nosocomial pulmonary fungal infection in patients with cancer are reducing risk factors, early diagnosis, and timely treatment.

Mycoses, 2004 Dec, 47(11-12), 524 - 6
A case of oral erosive candidosis in a kidney transplant patient; Romano C et al.; Summary A case of oral erosive candidosis due to Candida albicans in a 64-year-old female patient, who had undergone kidney transplant 20 days earlier, is reported . Concomitant herpes infection was excluded . The patient achieved clinical and mycological recovery after treatment with topical and systemic antimycotics (200 mg fluconazole per day) for 50 days . The case is reported because of the erosive ulcerating aspect and extent of the lesions, usually only reported in immunodepressed subjects, especially those with neutropenia or AIDS . Zusammenfassung Es wird uber eine orale, erosive Candidose durch Candida albicans bei einer 64-jahrigen Patientin berichtet . die 20 Tage zuvor eine Niere transplantiert bekommen hatte . Eine gleichizeitige Herpesinfektion wurde ausgeschlossen . Die Patientin war nacj 50 Tagen systemischer (200 mg Fluconazol taglich) und topischer Behandlung geheilt . Der Fall ist wegen des erosiven, ulzerierenden Aspekts und wegen des Schweregrades mitteilenswert, was gewohnlich nur bei immunsupprimierten Patienten mit Neutropenie oder bei AIDS gesehen wird.

Mycoses, 2004 Dec, 47(11-12), 482 - 90
Isolation and characterization of an immunogenic fragment of heat shock protein 60 from Trichophyton mentagrophytes; Raska M et al.; Summary Heat shock protein 60 (hsp60) were isolated from several fungal, protozoal and many bacterial pathogens and successfully used for protective vaccination in some infection models . This work focuses on the isolation of recombinant hsp60 from the dermatophyte, Trichophyton mentagrophytes as a potentially protective antigen in trichophytosis . With the help of a previously tested set of degenerated primers, it was used reverse transcriptase polymerase chain reaction (RT-PCR) for isolation of partial cDNA of the hsp60 T . mentagrophytes (labelled hsp60-TM814), which was cloned into cloning vector . The sequencing of hsp60-TM814 cDNA and global alignment confirmed homology of the hsp60-TM814 with other members of the hsp60 family . Hsp60-TM814 cDNA corresponds to the region encoding the immunoprotective fragment of the hsp60 from Histoplasma capsulatum, used successfully in mouse model of histoplasmosis . A recombinant fragment (r-hsp60-TM664), 220 amino acids in length, was prepared in a prokaryote expression system, and its identity confirmed by mass spectroscopy . High immunogenicity of r-hsp60-TM664 was proven after subcutaneous immunization of mice . Immunized mouse sera recognized r-hsp60-TM664 on Western blots as well as hsp60 from mouse liver lysate and lysate of Candida albicans . Zusammenfassung Hsp60-Proteine wurden aus verschiedenen pathogenen Pilzen, Protozoen und Bakterien isoliert und mit Erfolg fur protektive Vakzination in verschiedenen Infektionsmodellen benutzt . In unseren Untersuchungen konzentrierten wir uns auf die Isolierung des rekombinanten hsp60-Proteins aus dem Fadenpilz Trichophyton mentagrophytes als potenziell protektiven Antigens bei der Trichophytose . Mit Hilfe von vorher getesteten Primern benutzten wir die RT-PCR zur Isolierung einer partialen cDNA von T . mentagrophytes-hsp60 (bezeichnet hsp60-TM814), die in einen Klonierungsvektor kloniert wurde . Sequenzierung der hsp60-TM814-cDNA und globales Alignment bestatigten die Homologie von hsp60-TM814 mit anderen Genen der Hsp 60-Familie . Hsp60-TM814 entspricht der codierenden Region fur das immunogene, protektive Fragment des hsp60-Proteins von Histoplasma capsulatum, das erfolgreich im Mausemodell der Histoplasmose benutzt wurde . Ein rekombinantes Fragment (r-hsp60-TM664), 220 Aminosauren lang, wurde in einem prokaryotischen Expressionssystem synthetisiert und seine Identitat durch Massenspektroskopie bestatigt . Subkutane Immunisierung mit r-hsp60-TM664 bei Mausen hat seine hohe Immunogenitat nachgewiesen . An den Westernblots reagierte das Serum von immunisierten Mausen mit r-hsp60-TM664-Protein und weiter mit den hsp60-Proteinen aus den Lysaten von Mauseleber und dem Pilz Candida albicans.

Mycoses, 2004 Dec, 47(11-12), 465 - 469
Genotype distribution of Candida albicans isolates by 25S intron analysis with regard to invasiveness; Karahan ZC et al.; Summary The aim of this study was to genotype Candida albicans strains isolated from patients with invasive and non-invasive deep-seated infections . For this purpose, 301 C . albicans isolates (81 invasive and 220 non-invasive) were genotyped by using specific PCR primers designed to span the transposable group I intron of the 25S rDNA gene . Fifty-three of the 81 invasive isolates were genotype A (65.4%), eight were genotype B (9.9%) and 20 were genotype C (24.7%), while 98 of the 220 non-invasive isolates were genotype A (44.6%), 46 were genotype B (20.9%) and 76 were genotype C (34.5%) . Genotype A was more prevalent among invasive isolates and genotypes B and C were more prevalent among non-invasive isolates (P = 0.0046) . Genotypes D and E which represent C . dubliniensis were not found . These results indicate that there may be a relationship between C . albicans genotypes and invasiveness; genotype A being more invasive than others . The presence or absence of the transposable group I intron in the 25S rDNA gene may be important in determining the invasiveness of C . albicans . Zusammenfassung Ziel dieser Arbeit war, Candida albicans-Isolate von Patienten mit invasiven und nicht-invasiven Lasionen zu genotypisieren . Es wurden 301 Isolate (81 invasive und 220 nicht-invasive) untersucht . Die Genotypisierung wurde mittels eines spezifischen PCR Primers, der das Group I-Intron des 25S rDNA Gens umfasst, durchgefuhrt . 53 der invasiven Isolate waren Genotyp A (65,4%), 8 Genotyp B (9,9%), und 20 Genotyp C (24,7%); anderseits waren 98 der nicht-invasiven Isolate Genotyp A (44,6%), 46 Genotyp B (20,9%), und 76 Genotyp C (34,5%) . Genotyp A uberwog unter invasiven Isolaten, wahrend die Genotypen B und C unter nicht-invasiven Isolaten vorherrschten (P = 0.0046) . Die Genotypen D und E, die Candida dubliniensis umfassen, wurden nicht gefunden . Diese Ergebnisse zeigen, dass es eine Beziehung zwischen C . albicans-Genotypen und Invasivitat gibt und dass Genotyp A invasiver ist als andere . Das Group I-Intron des 25S rDNA Gens scheint fur die Invasivitatsabschatzung bei C . albicans brauchbar zu sein.

Int Rev Cytol, 2004, 242, 215 - 48
Multidrug resistance in yeast Candida; Prasad R et al.; The opportunistic human pathogens Candida albicans and other non-albicans species have acquired considerable significance in the recent past due to the enhanced susceptibility of immunocompromised patients . These pathogenic species of Candida derive their importance not only from the severity of their infections but also from their ability to develop resistance against antifungals . Widespread and prolonged use of azoles has led to the rapid development of the phenomenon of multidrug resistance (MDR), which poses a major hurdle in antifungal therapy . Various mechanisms that contribute to the development of MDR have been implicated in Candida as well as in other human fungal pathogens, and some of these include overexpression of or mutations in the target enzyme of azoles, lanosterol 14alpha-demethylase, and transcriptional activation of genes encoding drug efflux pump proteins belonging to ATP-binding cassette (ABC) as well as to major facilitator superfamilies (MFS) of transporters . The ABC transporters, CDR1, CDR2, and an MFS pump CaMDR1, play a key role in azole resistance as deduced from their high level of expression found in several azole-resistant clinical isolates.

Indian J Chest Dis Allied Sci, 2000 Oct-Dec, 42(4), 341 - 4
In vitro immunity of rat peritoneal macrophages to Candida albicans; Joshi KR et al.; Phagocytic, germ tube inducing and candidacidal activities were investigated in monolayers of peritoneal macrophages of rats . The phagocytic activities observed in macrophages of the healthy rats in the presence of normal serum, those in the presence of immune serum and of immunized rats in the presence of normal serum were 40%, 45.3% and 44.8% respectively . The percent of macrophages in which intracellular Candida formed germ tubes in the above three situations were 10, 9.59 and 10.19, respectively and the percent of intracellular Candida that formed germ tubes were 6.6, 3.7 and 4.1, respectively . The candidacidal activity observed in the above three sets of macrophages were 5.33%, 22.66% and 19.88%, respectively . Induction of germ tube in C . albicans in supplemented tissue culture medium containing normal serum was 15 per cent . These observations indicate that immunisation/sensitisation of individuals with C . albicans organisms does provide some degree of cell mediated immunity by activating macrophages . This may partly be due to the appearance of specific antibodies . It is likely that this type of immunity can be produced by subclinical infections during invasion by the commensal organism thus preventing further invasion establishment of infection and keeping the organism (C . albicans) in a state of commensalism . However, the degree of immunity so produced is so low that predisposing factors suppress it and allow establishment of infection.

Indian J Chest Dis Allied Sci, 2000 Oct-Dec, 42(4), 293 - 304
Paranasal sinus mycoses; Chakrabarti A et al.; The incidence of paranasal sinus mycoses (fungal sinusitis) varies widely with higher frequency in Sudan, southwestern states of USA and north India, which have hot and dry climate . The disease has been described as having four types: allergic, non-invasive, invasive and fulminate . A possible fifth type: non-invasive destructive may also exist . In a prospective study of 176 cases of fungal sinusitis from our centre, on the basis of clinical, radiological, histopathologic and mycologic findings the patients could be categorized into: allergic (12), non-invasive without bone destruction (81), non-invasive destructive (16), chronic invasive (55) and fulminant (12) types . Except the fulminate variety, the disease is commonly found in young immuno-competent population of rural areas . Aspergillus spp . are the commonest etiological agents though the importance of dematiaceous fungi in allergic fungal sinusitis has been stressed . Zygomycetes are common agents in fulminate type . In our series A . flavus (80%) was the commonest isolate, followed by A.fumigatus (9.7%), Rhizopus arrhizus (6.3%) and Alternaria spp . (1.1%) . Curvularia lunata, Apophysomyces elegans and Candida albicans were isolated from one patient each . Different host and environmental factors may help in lodging the causal fungi in mucosal plugs of these patients . Fungal allergy is associated with all varieties of the disease . But it is not clear what determines the invasion of mucosa . Rabbit can be used as an animal model . Histopathology and radio-imaging techniques help to distinguish different types and delineate extension of disease process . Culture helps to identify the responsible etiological agent . The presence or absence of precipitating antibody correlates well with disease progression or recovery . For effective management, non-invasive disease requires surgical debridement and sinus ventilation only . But for invasive type the need of adjuvant medical therapy is recommended to prevent recurrence and further extension . Itraconazole was found to be most useful in our study to prevent recurrence . Patients with fulminate type require radical surgery and immediate chemotherapy.

Biochemistry, 2004 Dec 21, 43(50), 15822 - 37
Structural features and thermodynamics of the J4/5 loop from the Candida albicans and Candida dubliniensis group I introns; Znosko BM et al.; The J4/5 loop of group I introns has tertiary interactions with the P1 helix that position the P1 substrate for the self-splicing reaction . The J4/5 loop of Candida albicans and Candida dubliniensis, 5'GAAGG3'/3'UAAUU5', potentially contains two A.A pairs flanked by one G.U pair on one side and two G.U pairs on the other side . Results from optical melting, nuclear magnetic resonance spectroscopy, and functional group substitution experiments with a mimic of the C . albicans and C . dubliniensis J4/5 loop are consistent with the adenosines forming tandem sheared A.A pairs with a cross-strand stack and only the G.U pair not adjacent to an A.A pair forming a static wobble G.U pair . The two G.U pairs adjacent to the tandem A.A pairs are likely in a dynamic equilibrium between multiple conformations . Although Co(NH(3))(6)(3+) stabilizes the loop by several kilocalories per mole at 37 degrees C, addition of Mg(2+) or Co(NH(3))(6)(3+) has no effect on the structure of the loop . The tandem G.U pairs provide a pocket of negative charge for Co(NH(3))(6)(3+) to bind . The results contribute to understanding the structure and dynamics of purine-rich internal loops and potential G.U pairs adjacent to internal loops.

J Med Microbiol, 2005 Jan, 54(Pt 1), 87 - 92
Divergent chemokine, cytokine and beta-defensin responses to gastric candidiasis in immunocompetent C57BL/6 and BALB/c mice; Schofield DA et al.; Previous studies of animal models of candidiasis have produced conflicting results concerning the cytokines and host defence mechanisms that are most relevant for protection against Candida infections . In this study, the host defence mechanisms evoked by two different immunocompetent murine strains following oral colonization with Candida albicans were assessed . beta-Defensin (mBD1, mBD3 and mBD4), chemokine (MIP-2 and KC) and cytokine (TNF-alpha, IFN-gamma, IL-4, IL-10, IL-12 and IL-15) gene expression in germ-free (gf) and C . albicans-infected (gastric) C57BL/6 and BALB/c mice was contrasted . Gf C57BL/6 and BALB/c mice expressed significantly different basal levels of mBD3, mBD4, TNF-alpha and IL-12 in gastric tissues; however, gf C57BL/6 and BALB/c mice were equally susceptible to intestinal colonization with C . albicans and had similar fungal burdens in gastric tissues 4 weeks after oral challenge . C57BL/6 mice responded to colonization and gastric candidiasis with increased expression of mBD1, mBD3, mBD4, TNF-alpha, MIP-2, KC and IL-12 . Conversely, a much more specific and attenuated response was observed in Candida-infected gastric tissues from BALB/c mice . Therefore, different strains of mice that were equally susceptible to gastric candidiasis after oral challenge had divergent cytokine, chemokine and beta-defensin responses . This suggests that conflicting data as to the relevance of cytokines and other host factors in murine resistance to candidiasis may be explained, at least in part, by the strain of mouse studied.

Eukaryot Cell, 2004 Dec, 3(6), 1639 - 52
TAC1, transcriptional activator of CDR genes, is a new transcription factor involved in the regulation of Candida albicans ABC transporters CDR1 and CDR2; Coste AT et al.; The ABC transporter genes CDR1 and CDR2 can be upregulated in Candida albicans developing resistance to azoles or can be upregulated by exposing cells transiently to drugs such as fluphenazine . The cis-acting drug-responsive element (DRE) present in the promoters of both genes and necessary for their upregulation contains 5'-CGG-3' triplets that are often recognized by transcriptional activators with Zn(2)-Cys(6) fingers . In order to isolate regulators of CDR1 and CDR2, the C . albicans genome was searched for genes encoding proteins with Zn(2)-Cys(6) fingers . Interestingly, three of these genes were tandemly arranged near the mating locus . Their involvement in CDR1 and CDR2 upregulation was addressed because a previous study demonstrated a link between mating locus homozygosity and azole resistance . The deletion of only one of these genes (orf19.3188) was sufficient to result in a loss of transient CDR1 and CDR2 upregulation by fluphenazine and was therefore named TAC1 (transcriptional activator of CDR genes) . Tac1p has a nuclear localization, and a fusion of Tac1p with glutathione S-transferase could bind the cis-acting regulatory DRE in both the CDR1 and the CDR2 promoters . TAC1 is also relevant for azole resistance, since a TAC1 allele (TAC1-2) recovered from an azole-resistant strain could trigger constitutive upregulation of CDR1 and CDR2 in an azole-susceptible laboratory strain . Transcript profiling experiments performed with a TAC1 mutant and a revertant containing TAC1-2 revealed not only CDR1 and CDR2 as targets of TAC1 regulation but also other genes (RTA3, IFU5, and HSP12) that interestingly contained a DRE-like element in their promoters . In conclusion, TAC1 appears to be the first C . albicans transcription factor involved in the control of genes mediating antifungal resistance.

Eukaryot Cell, 2004 Dec, 3(6), 1609 - 18
Snf7p, a component of the ESCRT-III protein complex, is an upstream member of the RIM101 pathway in Candida albicans; Kullas AL et al.; The success of Candida albicans as an opportunistic pathogen is based in part on its ability to adapt to diverse environments . The RIM101 pathway governs adaptation to neutral-alkaline environments and is required for virulence . Analysis of a genomic two-hybrid study conducted with Saccharomyces cerevisiae revealed that components involved in multivesicular bodies (MVB) transport may interact with RIM101 pathway members . Thus, we hypothesized that these proteins may function in the RIM101 pathway in C . albicans . We identified C . albicans homologs to S . cerevisiae Snf7p, Vps4p, and Bro1p and generated mutants in the cognate gene . We found that snf7Delta/Delta mutants, but not vps4Delta/Delta nor bro1Delta/Delta mutants, had phenotypes similar to, but more severe than, those of RIM101 pathway mutants . We found that the constitutively active RIM101-405 allele partially rescued snf7Delta/Delta mutant phenotypes . The vps4Delta/Delta mutant had subtle phenotypes, but these were not rescued by the RIM101-405 allele . Further, we found that the snf7Delta/Delta, vps4Delta/Delta, and bro1Delta/Delta mutants did not efficiently localize the vital dye FM4-64 to the vacuole and that it was often accumulated in an MVB-like compartment . This phenotype was not rescued by RIM101-405 or observed in RIM101 pathway mutants . These results suggest that Snf7p may serve two functions in the cell: one as a RIM101 pathway member and one for MVB transport to the vacuole.

Eukaryot Cell, 2004 Dec, 3(6), 1574 - 88
Deletion of the dynein heavy-chain gene DYN1 leads to aberrant nuclear positioning and defective hyphal development in Candida albicans; Martin R et al.; Cytoplasmic dynein is a microtubule-associated minus-end-directed motor protein . CaDYN1 encodes the single dynein heavy-chain gene of Candida albicans . The open reading frames of both alleles of CaDYN1 were completely deleted via a PCR-based approach . Cadyn1 mutants are viable but grow more slowly than the wild type . In vivo time-lapse microscopy was used to compare growth of wild-type (SC5314) and dyn1 mutant strains during yeast growth and after hyphal induction . During yeast-like growth, Cadyn1 strains formed chains of cells . Chromosomal TUB1-GFP and HHF1-GFP alleles were used both in wild-type and mutant strains to monitor the orientation of mitotic spindles and nuclear positioning in C . albicans . In vivo fluorescence time-lapse analyses with HHF1-GFP over several generations indicated defects in dyn1 cells in the realignment of spindles with the mother-daughter axis of yeast cells compared to that of the wild type . Mitosis in the dyn1 mutant, in contrast to that of wild-type yeast cells, was very frequently completed in the mother cells . Nevertheless, daughter nuclei were faithfully transported into the daughter cells, resulting in only a small number of multinucleate cells . Cadyn1 mutant strains responded to hypha-inducing media containing l-proline or serum with initial germ tube formation . Elongation of the hyphal tubes eventually came to a halt, and these tubes showed a defect in the tipward localization of nuclei . Using a heterozygous DYN1/dyn1 strain in which the remaining copy was controlled by the regulatable MAL2 promoter, we could switch between wild-type and mutant phenotypes depending on the carbon source, indicating that the observed mutant phenotypes were solely due to deletion of DYN1.

Eukaryot Cell, 2004 Dec, 3(6), 1423 - 32
KRE5 gene null mutant strains of Candida albicans are avirulent and have altered cell wall composition and hypha formation properties; Herrero AB et al.; The UDP-glucose:glycoprotein glucosyltransferase (UGGT) is an endoplasmic reticulum sensor for quality control of glycoprotein folding . Saccharomyces cerevisiae is the only eukaryotic organism so far described lacking UGGT-mediated transient reglucosylation of N-linked oligosaccharides . The only gene in S . cerevisiae with similarity to those encoding UGGTs is KRE5 . S . cerevisiae KRE5 deletion strains show severely reduced levels of cell wall beta-1,6-glucan polymer, aberrant morphology, and extremely compromised growth or lethality, depending on the strain background . Deletion of both alleles of the Candida albicans KRE5 gene gives rise to viable cells that are larger than those of the wild type (WT), tend to aggregate, have enlarged vacuoles, and show major cell wall defects . C . albicans kre5/kre5 mutants have significantly reduced levels of beta-1,6-glucan and more chitin and beta-1,3-glucan and less mannoprotein than the WT . The remaining beta-1,6-glucan, about 20% of WT levels, exhibits a beta-1,6-endoglucanase digestion pattern, including a branch point-to-linear stretch ratio identical to that of WT strains, suggesting that Kre5p is not a beta-1,6-glucan synthase . C . albicans KRE5 is a functional homologue of S . cerevisiae KRE5; it partially complements both the growth defect and reduced cell wall beta-1,6-glucan content of S . cerevisiae kre5 viable mutants . C . albicans kre5/kre5 homozygous mutant strains are unable to form hyphae in several solid and liquid media, even in the presence of serum, a potent inducer of the dimorphic transition . Surprisingly the mutants do form hyphae in the presence of N-acetylglucosamine . Finally, C . albicans KRE5 homozygous mutant strains exhibit a 50% reduction in adhesion to human epithelial cells and are completely avirulent in a mouse model of systemic infection.

Eukaryot Cell, 2004 Dec, 3(6), 1391 - 7
Role of Candida albicans transcription factor Upc2p in drug resistance and sterol metabolism; Silver PM et al.; In Candida albicans, drug resistance to clinically important antifungal drugs may be regulated through the action of transcription factors in a manner that may or may not be similar to regulation in Saccharomyces cerevisiae . A search of the C . albicans genome identified a single homolog of the S . cerevisiae transcription factor genes UPC2 (ScUPC2) and ECM22 (ScECM22) that have been associated with regulation of ergosterol biosynthesis . Sequence analysis of this C . albicans UPC2 (CaUPC2) gene identifies two domains, an anchoring transmembrane domain and a transcription factor region containing multiple nuclear localization signals and a fungal Zn(2)-Cys(6) binuclear cluster domain . Heterozygous deletion, homozygous deletion, and reconstructed strains of CaUPC2 as well as the parental strain were tested against several antifungal drugs, including ergosterol biosynthesis inhibitors . The CaUPC2 homozygous deletion strain showed marked hypersusceptibility to most drugs, compared to the parental and reconstructed strains . The deletion strains accumulate significantly less radiolabeled cholesterol, suggesting reduced ergosterol scavenging in those strains . When grown under azole drug pressure, the parental, heterozygous deletion and reconstructed strains of CaUPC2 upregulate the ERG2 and ERG11 ergosterol biosynthesis genes, while the homozygous deletion strain shows no such upregulation . Consistent with these results, CaUPC2 deletion strains show reduced ergosterol levels, which may explain the increased susceptibilities of the CaUPC2 deletion strains . Thus, it appears that CaUPC2 acts as a transcription factor involved in the regulation of ergosterol biosynthetic genes and as a regulator of sterol uptake across the plasma membrane.

J Ethnopharmacol, 2005 Jan 4, 96(1-2), 331 - 4
Biologically active traditional medicinal herbs from Balochistan, Pakistan; Zaidi MA et al.; The biological activities of the following four important medicinal plants of Balochistan, Pakistan were checked; Grewia erythraea Schwein f . (Tiliaceae), Hymenocrater sessilifolius Fisch . and C.A . Mey (Lamiaceae), Vincetoxicum stocksii Ali and Khatoon (Asclepiadaceae) and Zygophyllum fabago L . (Zygophyllaceae) . The methanolic extracts were fractionated into hexane, ethyl acetate, chloroform, butanol and water . The antifungal and antibacterial activities of these plants were determined against 12 fungal and 12 bacterial strains by agar well diffusion and disk diffusion assays . The extract of Zygophyllum fabago was found to be highly effective against Candida albicans and Escherichia coli . The extract of Vincetoxicum stocksii was also found to be significantly active against Candida albicans, Bacillus subtilis and Bacillus cereus . Extracts of Hymenocrater sessilifolius and Grewia erythraea showed good activity only against Pseudomonas aeruginosa.

J Ethnopharmacol, 2005 Jan 4, 96(1-2), 221 - 6
In vitro inhibitory effect of Streblus asper leaf-extract on adhesion of Candida albicans to human buccal epithelial cells; Taweechaisupapong S et al.; This in vitro study aimed at determining the effects of a sublethal concentration of Streblus asper Lour (Moraceae) leaf ethanolic extract on adherence of Candida albicans to human buccal epithelial cells (HBEC) . The minimum concentration of Streblus asper leaf ethanolic extract (SAE) that significantly reduced adherence (P<0.05) after a 1-h exposure was 15.6 mg/ml . However, there was a significant reduction (P<0.05) of candidal adhesion to HBEC after 1-min exposure to 125 mg/ml of SAE . Pre-treatment of either Candida or HBEC, or both, with 125 mg/ml of SAE for 1h resulted in reduced adherence . SAE at concentrations of 125 and 250 mg/ml also showed 41 and 61% inhibition of germ tube formation, respectively, which might affect adherence . These findings indicate that the sublethal concentration of SAE may modulate candidal colonization of the oral mucosa thereby suppressing the invasive potential of the pathogen.

Pharmazie, 2004 Nov, 59(11), 845 - 8
Allylthiosulfinate: beta-cyclodextrin inclusion complex: preparation, characterization and microbiological activity; Nikolic V et al.; An allylthiosulfinate: beta-cyclodextrin inclusion complex was synthesized and characterized using X-ray crystallography, IR spectroscopy, thermal analysis and nuclear magnetic resonance . The microbiological activity of the complex was tested on available fungi (Candida albicans ATCC 10231, Aspergillus niger ATCC 16404) and bacteria (Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 9027) . In small concentrations, the complex inhibited the growth of the microorganisms tested . The most susceptible microorganism was Candida albicans ATCC 10231, and the least susceptible Pseudomonas aeruginosa ATCC 9027.

J Nucl Med, 2004 Dec, 45(12), 2088 - 94
Synthesis and comparison of 99mTc-enrofloxacin and 99mTc-ciprofloxacin; Siaens RH et al.; The use of (99m)Tc-ciprofloxacin as a tracer for infection and inflammation has been examined and discussed in the literature extensively . Its alleged ability to discriminate between sterile inflammation and bacterial versus nonbacterial infections has led to an intense debate . Other labeled fluoroquinolones might offer better characteristics or may add to a better understanding of the working mechanism of (99m)Tc-ciprofloxacin . The rationale of this work was to determine possible differences in the use of 2 labeled quinolones--that is, (99m)Tc-ciprofloxacin and (99m)Tc-enrofloxacin--as tracers for infection and inflammation in animals . METHODS: Ciprofloxacin and enrofloxacin were labeled with (99m)Tc and characterized . The stability of both preparations was evaluated in serum and in the presence of an excess of cysteine . In vitro binding studies were performed to determine the interaction of the labeled quinolones with bacteria and other cells . Rats with sterile and infectious intramuscular lesions were used to study the scintigraphic properties of the 2 compounds . To assess the specificity of binding to living bacteria, infectious intramuscular lesions of heat-killed Staphylococcus aureus and Candida albicans were used as controls . Imaging was performed with a gamma-camera at 0, 3, 5, and 22 h after injection . RESULTS: The radiochemical purity of both radiolabeled fluoroquinolones exceeded 95% as determined by instant thin-layer chromatography . Both compounds were moderately stable in serum . Binding assays did not show any saturable binding to S . aureus, heat-killed S . aureus, as well as C . albicans . None of the tracers showed specific binding to bacteria . Scintigraphy showed uptake in the infectious lesion at 1 h after injection, which washed out during the next 4 h . Abscess-to-muscle ratios for both preparations were not significantly different for the various infectious or inflammatory lesions studied and did not exceed an average of 4.25 +/- 0.62 . CONCLUSION: The study demonstrated that (99m)Tc-ciprofloxacin and (99m)Tc-enrofloxacin do not show preferential binding to living bacteria . In vivo (99m)Tc-enrofloxacin has similar characteristics as (99m)Tc-ciprofloxacin except for differences in uptake in a few normal tissues.

Arh Hig Rada Toksikol, 2004 Nov, 55(4), 313 - 20
{Chemistry and biological effects of gliotoxin}; Kosalec I et al.; Gliotoxin is a mycotoxin from the epipolythiodioxypipeazine family with biological active internal disulfide bridge . Gliotoxin has an antibacterial and antiviral activity, but it was discarded from clinical practice due to its toxicity . The most studied effect of gliotoxin is its influence on the cell of the immune system . Today, researches are focused on treating transplantation organs ex situ and making them immunologically silent . Its toxicity has been proven on several cells (macrophages, thymocites, splenocytes, and fibroblasts) causing apoptosis and necrosis and it has acted as inhibitor of several enzymes (farnesyl-transefases, NF-kappaB, and alcohol-dehydrogenases) . Its mechanism of toxicity is connected with the production of mixed disulfide and covalent bonds, and oxidative effects . An important medical mould Aspergillus fumigatus and yeast Candida albicans can secrete gliotoxin in infected tissues and, because of the proven toxic effects of gliotoxin, it is suggested that gliotoxin can exacerbate mycoses (invasive aspergillosis or candidiasis) . Gliotoxin can also affect the invasiveness of fungi and their dissemination from the primary site throughout the organism.

J Clin Microbiol, 2004 Dec, 42(12), 5950 - 3
Refractory candidal meningitis in an immunocompromised patient cured by caspofungin; Liu KH et al.; Candidal meningitis is a rare infectious disease that usually leads to substantial morbidity and mortality . We present a case of candidal meningitis refractory to systemic antifungal therapy (amphotericin B and fluconazole) . A 63-year-old female with lymphoblastic lymphoma and myelodysplasia with leukemia transformation developed prolonged fever and headache on the seventh day following intrathecal prophylactic chemotherapy . A lumbar puncture showed neutrophilic pleocytosis, and a cerebrospinal fluid culture yielded Candida albicans . The clinical course was complicated by brain edema, subarachnoid hemorrhage, and hydrocephalus . Parenteral therapy with amphotericin B alone or amphotericin B in combination with fluconazole or intrathecal administration of amphotericin B failed to eradicate C . albicans in the cerebrospinal fluid . After 7 days of caspofungin therapy, however, the cerebrospinal fluid became sterile and the patient gradually regained consciousness . She was discharged 1 month after completing 4 weeks of caspofungin therapy . There were two critical issues we thought to be relevant to the favorable outcome of this case . First, isolation of C . albicans was achieved by inoculating enriched liquid medium with cerebrospinal fluid . Second, there is a potential therapeutic benefit of caspofungin in treating a fungal infection of the central nervous system.

J Clin Microbiol, 2004 Dec, 42(12), 5938 - 9
Molecular characterization of fluconazole resistance in a case of Candida albicans ocular infection; Pancholi P et al.; Ocular yeast infections in diabetics are a therapeutic challenge . Drug resistance and reduced azole susceptibility are major concerns . The case we describe characterizes a Candida albicans strain from a vitrectomy specimen that was susceptible to fluconazole by in vitro testing but recalcitrant to therapy . Molecular studies revealed transient overexpression of CDR1 and ERG11 mRNA in the presence of fluconazole that may have contributed to poor clinical response in this patient.

J Clin Microbiol, 2004 Dec, 42(12), 5899 - 903
Study of molecular epidemiology of candidiasis in portugal by PCR fingerprinting of Candida clinical isolates; Correia A et al.; PCR fingerprinting was used to type 177 yeast isolates obtained from two medical institutions . Candida albicans was the predominant species found, followed by C . tropicalis, C . glabrata, C . parapsilosis, C . guilliermondii, and C . krusei, which accounted for over 20% of the strains isolated . This survey represents the first study of molecular epidemiology of candidiasis in Portugal.

J Clin Microbiol, 2004 Dec, 42(12), 5624 - 35
Phylogeny and evolution of medical species of Candida and related taxa: a multigenic analysis; Diezmann S et al.; Hemiascomycetes are species of yeasts within the order Saccharomycetales . The order encompasses disparate genera with a variety of life styles, including opportunistic human pathogens (e.g., Candida albicans), plant pathogens (e.g., Eremothecium gossypii), and cosmopolitan yeasts associated with water and decaying vegetation . To analyze the phylogeny of medically important species of yeasts, we selected 38 human pathogenic and related strains in the order Saccharomycetales . The DNA sequences of six nuclear genes were analyzed by maximum likelihood and Bayesian phylogenetic methods . The maximum likelihood analysis of the combined data for all six genes resolved three major lineages with significant support according to Bayesian posterior probability . One clade was mostly comprised of pathogenic species of Candida . Another major group contained members of the family Metschnikowiaceae as a monophyletic group, three species of Debaryomyces, and strains of Candida guilliermondii . The third clade consisted exclusively of species of the family Saccharomycetaceae . Analysis of the evolution of key characters indicated that both codon reassignment and coenzyme Q(9) likely had single origins with multiple losses . Tests of correlated character evolution revealed that these two traits evolved independently.

Anal Biochem, 2005 Jan 1, 336(1), 39 - 45
The combination effects of phenolic compounds and fluconazole on the formation of ergosterol in Candida albicans determined by high-performance liquid chromatography/tandem mass spectrometry; Sun S et al.; A liquid chromatography/tandem mass spectrometry (LC/MS) with atmospheric pressure chemical ionization (APCI) for the quantification of ergosterol, lanosterol, and squalene was developed to evaluate the combination effects of phenolic compounds with fluconazole on ergosterol biosynthesis in Candida albicans . The three analytes were separated by a column of C18 and were quantified without interference with each other using positive mode tandem mass spectrometry (MS/MS) . Molecular ions of ergosterol and lanosterol were detected as the {M+H-H2O}+ ion species at m/z 380 and 410, whereas squalene appeared as the {M+H}+ ion species at m/z 412 . On fragmentation of ergosterol, lanosterol, and squalene, the product ions at m/z 69, 149, and 109, respectively, were present as major fragments . These product ions were used for the quantification of them in multiple reaction monitoring acquisition mode . The relationship between signal intensity and the analytes' concentration was linear over the concentration range of 0.05-10 microg/ml . Following the treatment of C . albicans with fluconazole in combination with albicanyl caffeate, resveratrol, and 3,4'-difluorostilbene, respectively, the content of ergosterol in both the sensitive and resistant C . albicans showed depletion, whereas the squalene showed accumulation especially in the sensitive isolates determined with the method developed.

J Antibiot (Tokyo), 2004 Sep, 57(9), 569 - 72
YM-193221, a novel antifungal antibiotic produced by Pseudallescheria ellipsoidea; Kamigiri K et al.; A novel antifungal antibiotic, YM-193221, was found in the culture broth of a fungus, Pseudallescheria ellipsoidea . The structure of the antibiotic was determined through several spectroscopic experiments as 2-dimethylamino-1-(4-hydroxyphenyl)-8,10-dimethyl-6-dodecene-3-one . YM-193221 exhibited potent antifungal activity against Candida albicans and also inhibited mannan synthesis in the yeast cell wall.

Matrix Biol, 2004 Nov, 23(7), 477 - 86
Basement membrane protein and matrix metalloproteinase deregulation in engineered human oral mucosa following infection with Candida albicans; Claveau I et al.; A variety of morphological changes in the basement membrane (BM) are known to occur in inflammatory diseases . Modifications of the BM can be associated with significant changes in protein content . Candida albicans (C . albicans) is normally a commensal organism and is a member of the natural flora of a large number of healthy individuals . However, under certain conditions, C . albicans can invade host tissues, causing inflammation and tissue damage . The aim of this study was to investigate the effect of C . albicans on the expression and production of structural (laminin-5 and type IV collagen) and inflammatory {matrix metalloproteinases (MMPs) and their inhibitors} proteins by human oral epithelial cells . Using engineered normal human oral mucosa infected with 10(5)C . albicans/cm(2) for different periods of time, we were able to demonstrate that this yeast promotes significant laminin-5 and type IV collagen gene activation and protein secretion . These effects were accompanied by MMP-2 and MMP-9 gene activation . Interestingly, only the levels of active MMP-9 rose . The increase in MMP levels was paralleled by a decrease in the secretion of type 2 matrix metalloproteinase tissue inhibitors (TIMP-2) . Our results demonstrated that C . albicans has a significant effect on tissue structure through BM protein and MMP modulation . This might help C . albicans overcome the mechanical and biological defenses of the tissue and allow it to disseminate, causing severe infections . If C . albicans uses MMPs (mainly MMP-9) to disseminate, inhibition of this protease could be of interest in treating a variety of inflammatory disorders, including oral candidiasis.

Arch Soc Esp Oftalmol, 2004 Nov, 79(11), 565 - 8
{Efficiency of voriconazole in fungal keratitis caused by candida albicans.}; Granados JM et al.; CASE REPORT: A 70-year-old woman, with a history of diabetes and persistent corneal ulceration, developed a severe keratitis caused by Candida albicans . It evolved rapidly to corneal perforation in spite of treatment with topical amphotericin B and oral itraconazole . An amniotic membrane transplant was performed as an emergency, associated with systemic administration of voriconazole . The infection was solved successfully, although the patient needed a penetrating keratoplasty to restore the corneal transparency . DISCUSSION: Fungal keratitis caused by Candida albicans usually follows an aggressive course and may simulate a bacterial aetiology . Voriconazole is a new antifungal drug that appears to be very effective in the management of ocular infections caused by many filamentous and levaduriform fungi (Arch Soc Esp Oftalmol 2004; 79: 565-568).

Plant Cell Physiol, 2004 Nov, 45(11), 1669 - 80
Pn-AMP1, a plant defense protein, induces actin depolarization in yeasts; Koo JC et al.; Pn-AMP1, Pharbitis nil antimicrobial peptide 1, is a small cysteine-rich peptide implicated in host-plant defense . We show here that Pn-AMP1 causes depolarization of the actin cytoskeleton in Saccharomyces cerevisiae and Candida albicans . Pn-AMP1 induces rapid depolarization of actin cables and patches within 15 min . Increased osmolarity or temperature induces transient actin depolarization and results in increased sensitivity to Pn-AMP1, while cells conditioned to these stresses show less sensitivity . Mutations in components of a cell wall integrity pathway (Wsc1p, Rom2p, Bck1p and Mpk1p), which regulate actin repolarization, result in increased sensitivity to Pn-AMP1 . A genetic screen reveals that mutations in components of the alpha-1,6-mannosyltransferase complex (Mnn10p, Mnn11p and Och1p), which regulate mannosylation of cell wall proteins, confer resistance to Pn-AMP1 . FITC-conjugated Pn-AMP1 localizes to the outer surface of the cell with no significant staining observed in spheroplasts . Taken together, these results indicate that cell wall proteins are determinants of resistance to Pn-AMP1, and the ability of a plant defense protein to induce actin depolarization is important for its antifungal activity.

Indian J Pediatr, 2004 Nov, 71(11), 973 - 7
Oral itraconazole in treatment of candidemia in a pediatric intensive care unit; Singhi SC et al.; OBJECTIVE: To examine efficacy of itraconazole in the treatment of candidemia in critically ill children . METHODS: We studied retrospectively cases of candidemia seen consecutively in our Pediatric Intensive Care Unit (PICU) over three and half years . Candida isolates from those patients included . Candida albicans--19, C . tropicalis--31, C . guillermondii--9, C.krusei--4 and C . glabrata--1 . RESULTS: Of the 64 patients, 48 (75%) had symptoms suggestive of septicemia and 16 had no symptoms suggestive of septicemia . No antifungal therapy was given to asymptomatic patients; they recovered from candidemia without development of any sequelae . Of the 48 symptomatic patients 11 died before results of fungal culture became available and antifungal therapy could be started . Thirty seven patients were treated with itraconazole (10 mg/kg/day orally or through gastric tube) . Seven (18.9 %) of 37 patients died, 3 within first week of antifungal therapy . Thirty (81%) patients recovered; microbiological cure was noted on average by day 14 (range 4-30 days) . The mean +/- SD duration of therapy in patients who responded was 24 +/-7 days (range 21-42 days) . None had any major side effect . CONCLUSION: We conclude that oral itraconazole may be effective in treatment of candidemia in children in a PICU where non-C . albicans candida species constituted majority (70%) of all Candida isolates.

Eur J Med Chem, 2004 Dec, 39(12), 1067 - 71
Synthesis and antifungal activity of cholesterol-hydrazone derivatives; Loncle C et al.; A series of hydrazones synthesized from various cholesterol derivatives were evaluated for their in vitro antimicrobial properties against human pathogens . The activity was highly dependent on the structure of the different compounds involved . The best results have been obtained with tosylhydrazone cholesterol derivatives 8 and 9 exhibiting activities against Candida albicans (CIP 1663-80) at a concentration of 1.5 microg/ml.

Acta Microbiol Immunol Hung, 2004, 51(3), 303 - 10
Innate immune functions of the keratinocytes . A review; Pivarcsi A et al.; Human keratinocytes are known to kill living microbes . They express different pattern recognition receptors (PRRs) such as the Toll-like receptor 2 (TLR2), TLR4, the CD1d molecule and a keratinocyte mannose-binding receptor (KcMR) . In response to challenge with microbes or microbial-derived substances the activation and nuclear translocation of NF-kappaB, the production of nitric oxide (NO) and inflammatory cytokines occur in keratinocytes, in a TLR-dependent manner . Blocking of NF-kappaB activation or NO production inhibit the Candida albicans-killing activity of keratinocytes . This Candida killing activity could be inhibited by blocking of KcMR . Recognition of invading pathogens in the epidermis triggers cytokine production in keratinocytes leading to elimination of pathogens and the activation of the adaptive immune system . These findings stress the importance of the role of keratinocytes in innate immunity.

J Nat Prod, 2004 Nov, 67(11), 1889 - 92
New macrolides from the sponge Chondrosia corticata; Shin J et al.; Three new oxazole-containing metabolites, neohalichondramide (5), (19Z)-halichondramide (6), and secohalichondramide (7), along with four previously reported compounds of the same structural class were isolated from the marine sponge Chondrosia corticata collected from Guam . The structures of novel compounds were determined on the basis of combined spectroscopic analyses . These compounds exhibited significant cytotoxicity and antifungal activity toward the human leukemia cell-line K562 and Candida albicans, respectively.

J Nat Prod, 2004 Nov, 67(11), 1879 - 81
(2S,3R)-2-aminododecan-3-ol, a new antifungal agent from the ascidian Clavelina oblonga; Kossuga MH et al.; A new antifungal agent, (2S,3R)-2-aminododecan-3-ol (1), has been isolated from the ascidian Clavelina oblonga collected in Brazil . The structure of 1 was established by analysis of spectroscopic data, including absolute stereochemistry determined by circular dichroism analysis of the dibenzoyl derivative 2 . Compound 1 displayed antifungal activity against Candida albicans ATCC 10231 with a MIC of 0.7 mug/mL and against Candida glabrata with a MIC of 30 microg/mL.

J Nat Prod, 2004 Nov, 67(11), 1809 - 17
New luffariellolide derivatives from the Indonesian sponge Acanthodendrilla sp; Elkhayat E et al.; Investigation of the Indonesian sponge Acanthodendrilla sp . afforded five new luffariellolide-related sesterterpenes, acantholides A-E (1-5), in addition to luffariellolide and its 25-O-methyl and 25-O-ethyl derivatives . All structures were unambiguously established by 1D and 2D NMR and MS spectroscopy . Acantholide D and E are derivatives comprising the 1-acetylcyclopentan-5-ol moiety, which are new variants of the C(14)-C(20) segment for this type of linear sesterterpenes . Luffariellolide and its 25-O-methyl congener as well as acantholide E (5) were cytotoxic against the mouse lymphoma L5187Y cell line . Acantholide B (2), luffariellolide, and its 25-O-methyl congener were active against the Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis, the Gram-negative bacterium Escherichia coli, the yeast Candida albicans, and the plant pathogenic fungus Cladosporium herbarum.

J Reprod Med, 2004 Oct, 49(10), 800 - 7
Activated lactoferrin and fluconazole synergism against Candida albicans and Candida glabrata vaginal isolates; Naidu AS et al.; OBJECTIVE: To evaluate the fungistatic activity of activated lactoferrin (ALF), fluconazole (FCN) individually and in combination against Candida vaginal isolates as well as to measure the time to recovery from the fungistatic effects after exposure in vitro to threshold minimal inhibitory concentrations (MIC) . STUDY DESIGN: Fungistasis patterns for ALF (2.5 mg/mL) and FCN (0.25 mg/mL) were tested at threshold MIC against vaginal isolates of C albicans (n = 5) and C glabrata (n = 5) grown in Sabouraud's dextrose broth against 10(5) yeast inoculum at 37 degrees C for 48 hours by microscale optical density (OD) assay according to the following criteria: "Total stasis" indicates that an agent elicited no change or a change in turbidity <0.1 OD unit for >48 hours (complete growth inhibition), "stasis recovery" (SR) is the time point at which turbidity of a previous stasis system shows an upward growth trend for >0.1 OD unit (recovery from growth inhibition), and "partial stasis" (PS) is proliferation after stasis recovery, measured as a percentage relative to growth control at any time (incomplete growth inhibition) . RESULTS: For ALF (2.5 mg/mL), the mean SR time was 15.6 +/- 2 hours for C albicans (n = 5) and 27.5 +/- 2 hours for C glabrata (n = 5) . The SR patterns for FCN were strain dependent and showed a wide range of deviation for both Candida species; accordingly, the values were 15.8 +/- 9 hours for C albicans and 25.5 +/- 12 hours for C glabrata . After 48 hours exposure to C albicans, ALF and FCN elicited a mean PS of 27.5 +/- 2% and 24.8 +/- 7%, respectively . The PS values at 48 hours showed a marked variation between C glabrata isolates, 29.1 +/- 24% for ALF and 21.5 +/- 38% for FCN . However, a combination of ALF and FCN at their threshold MIC showed significant drug synergism, causing total stasis of both species of Candida isolates . Thus, no SR for any Candida isolate was detected at or beyond 48 hours . Conversely, native lactoferrin failed to demonstrate such potent synergism with FCN against either Candida species . CONCLUSION: The combination of ALF and FCN at the threshold MIC elicited potent synergism, leading to total fungistasis of C albicans and C glabrata vaginal pathogens . ALF is a new class of fungistatic agent with a mode of action distinct from that of azoles.

Fitoterapia, 2004 Dec, 75(7-8), 768 - 70
Antimicrobial activity of crude methanolic extract of Satureja khuzistanica; Amanlou M et al.; The methanolic extract of the aerial parts of Satureja khuzistanica was investigated for its antimicrobial activity . The maximum antibacterial and antifungal activities were observed against Staphylococcus aureus and Candida albicans.

Fitoterapia, 2004 Dec, 75(7-8), 754 - 7
Antimicrobial and free radical scavenging activity of Gentianella nitida; Rojas R et al.; The antimicrobial and free radical scavenging activity of the ethanol extract and fractions of Gentianella nitida have been assessed . The most susceptible microorganisms were Candida albicans, Trichophyton mentagrophytes and Microsporum gypseum . The antifungal activity was concentrated in the 90% methanol and nonsoluble fractions, while the radical scavenging activity was stronger in the ethyl acetate and nonsoluble fractions.

Antimicrob Agents Chemother, 2004 Dec, 48(12), 4911 - 4
Need for early antifungal treatment confirmed in experimental disseminated Candida albicans infection; MacCallum DM et al.; Groups of mice infected intravenously with Candida albicans were treated intraperitoneally with amphotericin B, caspofungin, or fluconazole, starting at intervals before and after challenge . Survival was longest and tissue burdens were most reduced with early treatment, and survival times fell proportionately as treatment was delayed, reinforcing clinical recommendations for the earliest possible initiation of antifungal therapy.

Antimicrob Agents Chemother, 2004 Dec, 48(12), 4505 - 12
CaNdt80 is involved in drug resistance in Candida albicans by regulating CDR1; Chen CG et al.; Overexpression of CDR1, an efflux pump, is one of the major mechanisms contributing to drug resistance in Candida albicans . CDR1 p-lacZ was constructed and transformed into a Saccharomyces cerevisiae strain so that the lacZ gene could be used as the reporter to monitor the activity of the CDR1 promoter . Overexpression of CaNDT80, the C . albicans homolog of S . cerevisiae NDT80, increases the beta-galactosidase activity of the CDR1 p-lacZ construct in S . cerevisiae . Furthermore, mutations in CaNDT80 abolish the induction of CDR1 expression by antifungal agents in C . albicans . Consistently, the Candt80/Candt80 mutant is also more susceptible to antifungal drugs than the wild-type strain . Thus, the gene for CaNdt80 may be the first gene among the regulatory factors involved in drug resistance in C . albicans whose function has been identified.

Clin Dev Immunol, 2004 Sep-Dec, 11(3-4), 233 - 40
Plasmin promotes keratinocyte migration and phagocytic-killing accompanied by suppression of cell proliferation which may facilitate re-epithelialization of wound beds; Szabo I et al.; Keratinocytes were shown to induce the activation of plasminogen activator resulting in the formation of plasmin and the initiation of proteolysis in vitro . Activation of surface bound plasminogen may localize protease activity in the pericellular microenvironment and play a role in inducing both a conformational change and cell locomotion . Plasmin, however, can induce non-proteolytic effects on certain cell functions in a variety of cell lineages . In the present study we examined the effects of plasmin on keratinocytes with a focus on its role in the process of re-epithelialization, which included studies of cell migration, phagocytic-killing and cell proliferation . Migration of freshly isolated human epidermal keratinocytes was analyzed utilizing the agarose gel assay in the presence of 10% human serum . Plasmin at the concentration of 25 U/I induced a 160% increase in the chemotactic migration of keratinocytes that was completely blocked by the plasmin inhibitor alpha2-antiplasmin (Serpin) . In the absence of serum, plasmin also induced a reversible chemotactic migration of HaCaT keratinocytes as determined utilizing the microchemotaxis assay . Dose-response analysis showed a bi-phasic effect of plasmin with a maximum increase of 52% in keratinocyte chemotaxis at a concentration of 25 U/I . HaCaT cells on the other hand, showed no detectable in vitro chemokinesis by plasmin . Phagocytic-killing of Candida albicans by freshly isolated epidermal keratinocytes was enhanced in the presence of 25 U/I plasmin which was also reversible by the addition of Serpin . Spontaneous proliferation of HaCaT keratinocytes as determined by 3H-Thymidine uptake on the other hand, was reduced by 47 and 13% in cultures with 25 U/I plasmin for 24 and 48 h respectively, in a Serpin reversible manner . These data suggest that plasmin-induced chemotactic migration of epidermal keratinocytes is accompanied by enhanced phagocytic-killing coupled with suppression of proliferation of these cells which may facilitate re-epithelialization following skin injury.

J Enzyme Inhib Med Chem, 2004 Aug, 19(4), 373 - 9
Synthesis of coumarin derivatives with cytotoxic, antibacterial and antifungal activity; Khan KM et al.; The synthesis and selective biological screening of 7-hydroxy-4-methyl-2H-chromen-2-one (2), 7-hydroxy-4,5-dimethyl-2H-chromen-2-one (15) and some of their derivatives were carried out . Compound 13 was found to be most potent cytotoxic agent with LD50 = 126.69 microg/ml . In antibacterial assay the compounds showed a broad spectrum of activities . Compound 11 exhibited a very high degree of plant growth inhibition at three levels of concentration . Compound 4 showed very promising antifungal activity against Candida albicans . Compounds 12 and 13 demonstrated excellent antioxidant activity.

Infect Immun, 2004 Dec, 72(12), 7330 - 3
The calcineurin target, Crz1, functions in azole tolerance but is not required for virulence of Candida albicans; Onyewu C et al.; In Candida albicans, calcineurin is essential for virulence and survival during membrane perturbation by azoles . Crz1 is a proposed downstream target of calcineurin based on studies of Saccharomyces cerevisiae . However, the in vitro phenotypes of C . albicans crz1/crz1 and calcineurin mutants differ and Crz1 is not required for virulence.

FEMS Yeast Res, 2004 Dec, 5(3), 287 - 96
Transcriptional profiling of the early stages of germination in Candida albicans by real-time RT-PCR; Toyoda M et al.; By using real-time RT-PCR, we profiled the expression of CGR1, CaMSI3, EFG1, NRG1, and TUP1 in Candida albicans strains JCM9061 and CAI4 under several conditions, including induction of morphological transition, heat shock, and treatment with calcium inhibitors . Expression of CaMSI3 changed under these growth conditions except during heat shock . CGR1 expression increased during the early stages of hyphal growth in JCM9061, while expression was strain-dependent during heat shock . Both EFG1 and NRG1 were similarly expressed under hypha-inducing conditions and heat shock . Expression of TUP1 was slightly different from the expression of EFG1 or NRG1.

Curr Opin Microbiol, 2004 Dec, 7(6), 661 - 5
Through a glass opaquely: the biological significance of mating in Candida albicans; Magee PT et al.; Most Candida albicans strains are heterozygous at the MTL (mating-type-like) locus, but mating occurs in hemi- or homozygous strains . The white-opaque switch process is repressed by the heterodimer of the MTLa1 and MTLalpha2 gene products, while mating genes are induced by a2 and alpha1 . Mating occurs in opaque cells and produces tetraploid progeny . A small percentage (3-7%) of clinical isolates are homozygous at the MTL locus and most are mating-competent . MTL gene expression is controlled in part by a gene which activates MTLalpha genes and represses MTLa genes in response to hemoglobin . A failure to find meiosis and the lack of evidence of mating in vivo, together with some of the properties of opaque cells, leads to the suggestion that mating may have persisted because the tightly associated switch facilitates the commensal lifestyle of this fungus.

Mol Microbiol, 2004 Dec, 54(5), 1335 - 51
Transcriptional profiling in Candida albicans reveals new adaptive responses to extracellular pH and functions for Rim101p; Bensen ES et al.; The human pathogen Candida albicans grows and colonizes sites that can vary markedly in pH . The pH response in C . albicans is governed in part by the Rim101p pathway . In Saccharomyces cerevisiae, Rim101p promotes alkaline responses by repressing expression of NRG1, itself a transcriptional repressor . Our studies reveal that in C . albicans, Rim101p-mediated alkaline adaptation is not through repression of CaNRG1 . Furthermore, our studies suggest that Rim101p and Nrg1p act in parallel pathways to regulate hyphal morphogenesis, an important contributor to virulence . To determine the wild-type C . albicans transcriptional response to acidic and alkaline pH, we utilized microarrays and identified 514 pH-responsive genes . Of these, several genes involved in iron acquisition were upregulated at pH 8, suggesting that alkaline pH induces iron starvation . Microarray analysis of rim101-/- cells indicated that Rim101p does not govern transcriptional responses at acidic pH, but does regulate a subset of transcriptional responses at alkaline pH, including the iron acquisition genes . We found that rim101-/- cells are sensitive to iron starvation, which suggests that one important aspect of the Rim101p-dependent alkaline pH response is to adapt to iron starvation conditions.

Mol Microbiol, 2004 Dec, 54(5), 1212 - 23
A Pseudomonas aeruginosa quorum-sensing molecule influences Candida albicans morphology; Hogan DA et al.; Candida albicans is an opportunistic pathogen that is commonly found as a member of the human microflora . The ability of C . albicans to alter its cellular morphology has been associated with its virulence; yeast cells are more prevalent in commensal interactions whereas filamentous cells appear important in opportunistic infections . C . albicans encounters a multitude of other microbial species in the host environment and it is likely that they impact the C . albicans transition between virulent and non-virulent states . Here, we report that C . albicans morphology is significantly affected by the presence of Pseudomonas aeruginosa, another opportunistic pathogen . In a screen using a C . albicans HWP1-lacZ strain to indicate regions of filamentous growth, we identified P . aeruginosa mutants incapable of inhibiting C . albicans filamentation . Through these studies, we found that 3-oxo-C12 homoserine lactone, a cell-cell signalling molecule produced by P . aeruginosa, was sufficient to inhibit C . albicans filamentation without affecting fungal growth rates . Both microscopic analysis and real-time reverse transcription polymerase chain reaction analysis of morphology-specific markers confirmed that filamentation was suppressed by 200 microM 3-oxo-C12 homoserine lactone . Structurally related compounds with a 12-carbon chain length, e.g . C12-acyl homoserine lactone and dodecanol also affected C . albicans filamentation at similar concentrations . In contrast, other acylated homoserine lactones of different chain lengths did not affect fungal morphology . The activity of 3OC12HSL is compared with that of farnesol, a C . albicans-produced molecule also with a C12-backbone . The effects that bacteria have on the morphology of C . albicans represents one of the ways by which bacteria can influence the behaviour of fungal cells.

Zh Mikrobiol Epidemiol Immunobiol, 2004 Sep-Oct, (5), 80 - 4
{Reactivity of Candida albicans in the system of the alternative way of complement activation}
{Influence of staphylococcal metabolites on adhesive reactions in the system "Candida albicans-buccal epitheliocytes"}
{No authors listed}

The influence of S . aureus and S . epidermitidis metabolites on the adhesive reactions in the system "C . albicans-buccal epitheliocytes" was studied . The study revealed that the treatment of C . albicans with S . aureus supernatants inhibited the adhesion of C . albicans to epitheliocytes, the degree of the inhibiting action of S . aureus supernatants in the system depending on their strain specificity . S . epidermitidis supernatants produced no adhesive effect . The irreversible decrease of the adhesive activity of C . albicans under the action of bacterial metabolites was, seemingly, the consequence of transformation of the receptor apparatus of C . albicans . At the same time S . aureus supernatants produced no essential influence on the adhesive potential and viability of buccal epitheliocytes.

Med Mycol, 2004 Oct, 42(5), 461 - 73
Isolation, characterization, and regulation of the Candida albicans ERG27 gene encoding the sterol 3-keto reductase; Pierson CA et al.; The Candida albicans ERG27 gene which encodes the 3-keto reductase enzyme required for sterol C-4 demethylation was isolated and found to encode a 349 amino acid protein that is 60% identical at the amino acid level to the Saccharomyces cerevisiae Erg27p . A C . albicans erg27 null was created in a strain containing an integrated ERG27 rescue cassette under the control of the pMAL2 inducible promoter . The C . albicans erg27 strain was able to grow only in the presence of maltose indicating that the ERG27 gene is essential . The C . albicans erg27 null showed complete loss of both 3-keto reductase and oxidosqualene cyclase (Erg7p) activities compromising all sterol synthesis . These results suggest that Erg27p inhibitors might be effective antifungals . To explore ERG27 regulation, an erg11 null strain was generated . C . albicans erg6 and erg24 mutants were also employed along with the inhibitors, itraconazole and zaragozic acid A, to characterize ERG27 expression using Northern analysis . Expression was increased two- to fourfold in erg11, erg6 and erg24 backgrounds . However, itraconazole which targets Erg11p (lanosterol demethylase) increased ERG27 expression 10-fold and zaragozic acid A which targets the Erg9p (squalene synthase) increased ERG27 expression fivefold . The azole and erg11 results support other observations that azoles may affect non-sterol targets.

Med Mycol, 2004 Oct, 42(5), 439 - 47
Candida albicans PLD I activity is required for full virulence; Dolan JW et al.; Phospholipase D1 (PLD1) mutants of Candida albicans are defective in important in vivo and in vitro virulence factors . PLD1 mutants colonize the murine alimentary tract as well as PLD1 sufficient strains . In comparison to PLD1 sufficient strains, the PLD1 mutants: (i) are unable to survive in internal organs after intravenous challenge; (ii) do not decrease the body weights of mice after oral challenge; and (iii) are not lethal for immunodeficient mice after oral challenge . In vitro, the PLD1 mutants show a drastically reduced capacity to penetrate epithelial monolayers and they fail to develop hyphae when grown on solid Spider medium . The morphogenic switch from yeast to hyphae is controlled by multiple parallel signaling pathways that couple specific stimuli to the regulation of several transcription factors . Our data suggest that PLD1 functions in at least one of these pathways regulating morphogenesis in vitro and that while the mutants are able to form hyphae in vivo, the hyphae are defective in their ability to cause oroesophageal and gastric candidiasis and to kill the C . albicans-colonized mice.

Med Mycol, 2004 Oct, 42(5), 427 - 32
Chemiluminescent visualization of superoxide generated by Candida albicans; Masui S et al.; The high toxicity of reactive oxygen species (ROS) suggested a possible role in the pathogenicity of human pathogenic fungi . We previously reported a chemiluminescence method for measuring ROS generation in Candida albicans . In the present study, we attempted to visualize the ROS, superoxide anion radical (O2-), generated by paraquat (PQ)-stimulated C . albicans using methyl-Cypridina-luciferin analog (MCLA) as a chemiluminescence probe . Colonies of a wild-type C . albicans parent strain and its respiration-deficient mutant grown on agar plates were overlaid with a mixture of PQ and MCLA solutions . MCLA-dependent light emission from the colonies was recorded with a Hamamatsu ultralow-light-imaging apparatus with a CCD camera in a light-tight box . In the wild-type strain, marginal regions of growing colonies were strongly illuminated . The light emission from the colonies was extinguished by superoxide dismutase (SOD), proving that the light emission was strictly due to the superoxide anion . However, colonies of the respiration-deficient mutant poorly generated superoxide . Chemiluminecence measurements by a luminometer showed vigorous superoxide generation by the exponential phase cells of the parent strain but weak generation by the stationary phase cells . In the mutant, superoxide generation was weak compared with the parent strain . These results indicate that expansion of the colonies was due to the actively respiring cells located in the marginal regions . To our knowledge, the present report is the first chemiluminescent visualization of ROS including superoxide generated by C . albicans.

Nippon Ishinkin Gakkai Zasshi, 2004, 45(4), 233 - 7
{Clinical and fundamental investigations on recurrent glossodynia.}; Satoh T; A type of oral lesion, so-called glossodynia has been on the increase recently . Glossodynia is a kind of psychosomatic disease in which the patient experiences chronic pain on the surface of the tongue . It has never been diagnosed as coming from organic or functional pain . Although glossodynia can be cured by antianxiety drugs, antidepressants, or autogenic training and so on, usually . These are not a desirable solution . We initially tried to administer the antifungal drug, ITCZ, to 65 glossodynia patients . Sixty-four of them were cured of tongue pain after 1-3 weeks . The effective rate of recovery was 98.5% . Only two patients experienced reccurence of pain after 15 and 17 weeks, respectively, and Candida albicans was isolated from the surface of their tongues . The nature of the recurrent strains was investigated by MICs against 4 antifungal drugs, ITCZ, MCZ, AMPH-B, and NYS, as well as by the API 20C Auxanogram biochemically, and a molecular epidemiologic study by PFGE . Each case of Candida albicans was almost the same before and after the administration of ITCZ . Above all, it is important to carefully inspect the candidiasis of the tongue and to initially administer antifungal drugs when the diagnosis is glossodynia.

Nippon Ishinkin Gakkai Zasshi, 2004, 45(4), 227 - 31
{Development of murine experimental model for candidiasis and its application.}; Abe S; Candida albicans is a major cause of oral and esophageal infections in elder persons with poor oral hygiene and immuncompromised patients with hyposalivation, diabetes mellitus, prolonged use of antibiotics or immunosuppressive drugs . Oral thrush is a common form of oropharyngeal candidiasis whose clinical features consist of white patches appearing as discrete lesions on the buccal mucosa, throat, tongue, and gum linings that develop into confluent pseudomembranes resembling milk curds . We recently reported a simple murine model of thrush-type oral candidiasis that mimics the natural infectionin humans and is useful for both symptomatological and mycological evaluation of the responsiveness to antifungal treatments . By using this oral candidiasis model, protective activities of oral administration of several types of herbal preparations such as teatree oil, clove preparation and bovine lactoferrin were clarified . The mechanisms of protective actions of lactoferrin against oral candidiasis were particularly elucidated to include augmentation of T-cell activities of lesional lymphoid tissues . More recent studies suggested that saliva from healthy persons also shows a protective action for this murine oral candidiasis model.

Ann Pharm Fr, 2004 Nov, 62(6), 428 - 30
{Antimicrobial activity of new 4,4'-bipyridine derivatives: and in vitro study}; Rotaru A et al.; The antimicrobial and antifungal activity of some 4,4'-bipyridine derivatives were studied . The diffusimetric gelose surface diffusion method with stainless steel cylinders was used to study bacteria and Sabouraud fields for Candida albicans . Comparative analysis of the results led to the following conclusions . Diquaternary salts of 4,4'-bipyridinium possess a remarkable antimicrobial and antifungal activity . The influence of R1 and R2 substitutes in the para or meta position of the benzoylic radical affects selectivity but does not greatly influence activity.

Curr Genet, 2004 Dec, 46(6), 343 - 56 Epub 2004 Nov 10.
Random mutagenesis of an essential Candida albicans gene; Palmer GE et al.; A method for the analysis of Candida albicans gene function, which involves random mutagenesis of the open reading frame, is described . This method is especially suited for the study of essential and multi-functional genes, with several advantages over regulatable promoters more commonly used to study essential gene function . These advantages include expression from the endogenous promoter, which should yield a more appropriate transcript expression and abrogate the need for shifts in carbon or amino acid sources necessary with the use of regulatable promoters . Furthermore, there is potential for isolating individual functions of multi-functional genes . To verify this experimental approach, we randomly mutated the essential C . albicans gene, BMH1 . The resulting "pool" of putative mutant alleles was then introduced into a BMH1/bmh1Delta strain of C . albicans, such that only the mutagenized BMH1 sequences could be expressed . Transformants were screened for rapamycin sensitivity, defects in filamentation on M199 agar, and growth at 42 degrees C . In this way, we identified six non-lethal mutant alleles of BMH1 with altered amino acid sequences . Further phenotypic analysis of these mutant strains enabled us to segregate individual functions of C . albicans BMH1 . The relative merits of Escherichia coli versus PCR-mediated mutagenesis are discussed.

J Appl Microbiol, 2004, 97(6), 1201 - 9
Penicillium chrysogenum glucose oxidase -- a study on its antifungal effects; Leiter E et al.; AIMS: Purification and characterization of the high molecular mass Candida albicans-killing protein secreted by Penicillium chrysogenum . METHODS AND RESULTS: The protein was purified by a combination of ultrafiltration, chromatofocusing and gel filtration . Enzymological characteristics {relative molecular mass (M(r)) = 155 000, subunit structure alpha(2) with M(r,alpha) = 76 000, isoelectric point (pI) = 5.4} were determined using SDS-PAGE and 2D-electrophoresis . N-terminal amino acid sequencing and homology search demonstrated that the antifungal protein was the glucose oxidase (GOX) of the fungus . The enzyme was cytotoxic for a series of bacteria, yeasts and filamentous fungi . Vitamin C (1.0 mg ml(-1)) prevented oxidative cell injuries triggered by 0.004 U GOX in Emericella nidulans cultures but bovine liver catalase was ineffective even at a GOX : catalase activity ratio of 0.004 : 200 U . A secondary inhibition of growth in E . nidulans cultures by the oxygen-depleting GOX-catalase system was likely to replace the primary inhibition exerted by H(2)O(2) . CONCLUSIONS: Penicillium chrysogenum GOX possesses similar enzymological features to those described earlier for other Penicillium GOXs . Its cytotoxicity was dependent on the inherent antioxidant potential of the test micro-organisms . SIGNIFICANCE AND IMPACT OF THE STUDY: Penicillium chrysogenum GOX may find future applications in glucose biosensor production, the disinfection of medical implants or in the food industry as an antimicrobial and/or preservative agent.

Farmaco, 2004 Nov, 59(11), 863 - 8
Azaanalogues of ebselen as antimicrobial and antiviral agents: synthesis and properties; Wojtowicz H et al.; The different analogues of ebselen-unsubstituted benzisoselenazol-3(2H)-one (2a) 2-pyridylbenzisoselenazol-3(2H)-ones (2b-h) and 7-azabenzisoselenazol-3(2H)-ones (3a-j) were designed as new selenium-containing antiviral and antimicrobial agents and synthesized . Some of them were found in the antiviral assay in vitro to be strong inhibitors of cythopatic activity of herpes simplex virus type 1--HSV-1 (compounds 2a,b,f,h, 3a-j) and encephalomyocarditis virus--EMCV (compounds 2a,h, 3a-f,k,l) . The compounds 2a,h and 3a-e,j were found to have an appreciable activity against Gram-positive bacteria (Staphylococcus aureus and Bacillus) in vitro, some of them inhibited growth of pathogenic yeasts (Candida albicans) (3a,b) and filamentous fungi (3a-e,f).

Yeast, 2004 Nov, 21(15), 1269 - 77
Allelic isoforms of the H+/nucleoside co-transporter (CaCNT) from Candida albicans reveal separate high- and low-affinity transport systems for nucleosides; Slugoski MD et al.; Contigs 19-10196 and 19-20196 of the Stanford Candida albicans genome sequence databank encode two putative allelic isoforms of C . albicans CaCNT, a recently characterized 608 amino acid residue H+-coupled fungal member of the CNT family of concentrative nucleoside transport proteins . The single Ser/Gly difference between CaCNT/19-20196 and CaCNT occurs at position 328 in putative TM 7, and corresponds to a Ser/Gly substitution previously shown to contribute to the contrasting pyrimidine and purine nucleoside selectivities of human (h) and rat (r) Na+-dependent CNT1 and CNT2 . CaCNT/19-10196 differs from CaCNT by four amino acids, but has Gly at position 328 . These new proteins were recreated by site-directed mutagenesis of CaCNT and characterized functionally by heterologous expression in Xenopus laevis oocytes . In marked contrast to h/rCNT1/2, both CaCNT/19-10196 and CaCNT/19-20196 exhibited permeant selectivities for purine nucleosides (adenosine, guanosine and inosine) and uridine similar to that of CaCNT . However, although H+-coupled, CaCNT/19-20196 exhibited a approximately 10-fold higher apparent Km for uridine than either CaCNT or CaCNT/19-10196 . CaCNT/19-20196 also exhibited a low apparent affinity for inosine . We conclude that the three proteins correspond to high-affinity (CaCNT, CaCNT/19-10196) and low-affinity (CaCNT/19-20196) allelic isoforms of the C . albicans CNT nucleoside transporter . This is the first example of a single amino acid residue substitution altering a CNT protein's overall apparent affinity for nucleosides.

Eur Cytokine Netw, 2004 Jul-Sep, 15(3), 263 - 71
Myeloid differentiation factor 88 (MyD88) is required for murine resistance to Candida albicans and is critically involved in Candida -induced production of cytokines; Villamon E et al.; We have studied the role of myeloid differentiation factor 88 (MyD88), the universal Toll-like receptor (TLR) adaptor protein, in murine defenses against Candida albicans . MyD88-deficient mice, experimentally infected in vivo, had a very significant impaired survival, and a higher tissue fungal burden when compared with control mice . The recruitment of neutrophils to the site of infection was also significantly diminished in MyD88-\- mice . In vitro production of proinflammatory cytokines such as TNF-alpha, IFN-gamma and IL-12p70, by antigen-stimulated splenocytes from mice intravenously infected with the low-virulence C . albicans PCA2 strain, could not be detected in MyD88-\- mice . This default of production of Th1 cytokines in MyD88-deficient mice correlated with a greatly diminished frequency of IFN-gamma-producing CD4 + T lymphocytes . Also, the frequency of IFN-gamma-producing CD8 + T lymphocytes was lower in MyD88-\- mice than in control mice . Although C . albicans-specific antibody titers in PCA2-infected mice appeared more quickly in MyD88-\- mice than in control mice, the MyD88-\- group was not able to maintain the Candida-specific IgM nor IgG titers at the third week of infection . The complexity of antigens recognized by sera from MyD88-\- mice was quite similar to that from infected control mice . Taken together, these data show that MyD88-\- mice are extremely susceptible to C . albicans infections, suggesting that MyD88-dependent signaling pathways are essential for both the innate and adaptive immune responses to C . albicans.

Diagn Microbiol Infect Dis, 2004 Nov, 50(3), 187 - 92
In vitro fluconazole susceptibility of 1565 clinical isolates of candida species evaluated by the disk diffusion method performed using NCCLS M44-A guidelines; Testore GP et al.; We determined the in vitro activity of fluconazole against 1565 clinical Candida spp . isolates collected from different specimens of non-AIDS outpatients and inpatients in 3 different regions of Italy . Susceptibility testing was performed by agar disk diffusion using the NCCLS document M44-A guidelines . Candida albicans was the most frequently isolated yeast (68%) followed by C . glabrata (15%), C . tropicalis (5%), C . parapsilosis (5%), and C . krusei (5%) . Other yeasts represented 4% of all isolates . Of the 1565 isolates tested, 1449 (92.6%) were susceptible (S) to fluconazole, 43 (2.7%) were susceptible dose-dependent (S-DD) and 73 (4.7%) were resistant (R) . Almost all (98.2%) of the C . albicans isolates were classified as S or S-DD . Despite its widespread use, fluconazole displayed good activity against the isolates we tested, and the disk diffusion method was confirmed as a reliable approach to the evaluation of in vitro susceptibility of yeasts to this antimycotic agent.

Clin Diagn Lab Immunol, 2004 Nov, 11(6), 1111 - 9
Regulation of fungal infection by a combination of amphotericin B and peptide 2, a lactoferrin peptide that activates neutrophils; Okamoto T et al.; To establish a novel strategy for the control of fungal infection, we examined the antifungal and neutrophil-activating activities of antimicrobial peptides . The duration of survival of 50% of mice injected with a lethal dose of Candida albicans (5 x 10(8) cells) or Aspergillus fumigatus (1 x 10(8) cells) was prolonged 3 to 5 days by the injection of 10 microg of peptide 2 (a lactoferrin peptide) and 10 microg of alpha-defensin 1 for five consecutive days and was prolonged 5 to 13 days by the injection of 0.1 microg of granulocyte-monocyte colony-stimulating factor (GM-CSF) and 0.5 microg of amphotericin B . When mice received a combined injection of peptide 2 (10 microg/day) with amphotericin B (0.5 microg/day) for 5 days after the lethal fungal inoculation, their survival was greatly prolonged and some mice continued to live for more than 5 weeks, although the effective doses of peptide 2 for 50 and 100% suppression of Candida or Aspergillus colony formation were about one-third and one-half those of amphotericin B, respectively . In vitro, peptide 2 as well as GM-CSF increased the Candida and Aspergillus killing activities of neutrophils, but peptides such as alpha-defensin 1, beta-defensin 2, and histatin 5 did not upregulate the killing activity . GM-CSF together with peptide 2 but not other peptides enhanced the production of superoxide (O2-) by neutrophils . The upregulation by peptide 2 was confirmed by the activation of the O2- -generating pathway, i.e., activation of large-molecule guanine binding protein, phosphatidyl-inositol 3-kinase, protein kinase C, and p47phox as well as p67phox . In conclusion, different from natural antimicrobial peptides, peptide 2 has a potent neutrophil-activating effect which could be advantageous for its clinical use in combination with antifungal drugs.

Rev Iberoam Micol, 2004 Jun, 21(2), 70 - 4
Differences in extracellular enzymatic activity between Candida dubliniensis and Candida albicans isolates; Vidotto V et al.; Twenty-six Candida dubliniensis and 27 Candida albicans oral strains isolated from patients infected by the human immunodeficiency virus (HIV) were tested for germ tube production and 21 extracellular enzymatic activities . Assessment of the enzymatic profile was performed by using the API-ZYM commercial kit system (bioMerieux, France), which tests 19 different enzymes . Protease activity was expressed during the first days of incubation by 100% of the strains studied and resulted higher than phospholipase activity in the C . dubliniensis and C . albicans strains tested . The API-ZYM profile of the C . dubliniensis and C . albicans strains differs with respect to the number and percentage of the enzymes considered, as well as with the intensity of the substrate metabolized by the strains, in particular for the enzymes n 8 (cystine-arylamidase), n 12 (naphtol-AS-BI-phosphohydrolase) and n 16 (alpha-glucosidase) . These enzymes may be useful to differentiate C . dubliniensis and C . albicans together with other phenotypic characteristics proposed in the literature . No relationship among protease, phospholipase and other extracellular enzymatic activities was observed in C . dubliniensis . The average percentage of strains filamentation after 4 h was between 32 and 42%.

Rev Iberoam Micol, 1997 Dec, 14(4), 150 - 4
{Molecular and genetic aspects of resistance to azoles in Candida albicans.}; Hernaez ML et al.; Fluconazole is one of the most useful drugs in the treatment of fungal systemic infections which frequently affect non immunocompetent individuals . However, the emergence of resistant strains in recent years may severely limit its usefulness in future . Although there are several described mechanisms involved in resistance to azoles, recent genetic studies demonstrate the role of specific genes in clinical resistance . Currently, the best characterized are the MDR1 and CDR1 genes, which code members of the MFS or ABC familiy of drug transporters, respectively . These proteins respond to the membrane potential (MFS) or hydrolyse ATP (ABC) thus promoting drug efflux and therefore reducing its intracellular accumulation . It has been shown that the mRNA from these genes is frequently increased in some Candida albicans resistant strains from patients receiving long term azole treatment . The development of molecular genetic tools in C . albicans is allowing characterization of their role in this and other important processes in the fungal cell.

PLoS Biol . 2004 Dec;2(12):e398 . Epub 2004 Nov 09.
Conservation and evolution of cis-regulatory systems in ascomycete fungi; Gasch AP et al.; Relatively little is known about the mechanisms through which gene expression regulation evolves . To investigate this, we systematically explored the conservation of regulatory networks in fungi by examining the cis-regulatory elements that govern the expression of coregulated genes . We first identified groups of coregulated Saccharomyces cerevisiae genes enriched for genes with known upstream or downstream cis-regulatory sequences . Reasoning that many of these gene groups are coregulated in related species as well, we performed similar analyses on orthologs of coregulated S . cerevisiae genes in 13 other ascomycete species . We find that many species-specific gene groups are enriched for the same flanking regulatory sequences as those found in the orthologous gene groups fromS . cerevisiae, indicating that those regulatory systems have been conserved in multiple ascomycete species . In addition to these clear cases of regulatory conservation, we find examples of cis-element evolution that suggest multiple modes of regulatory diversification, including alterations in transcription factor-binding specificity, incorporation of new gene targets into an existing regulatory system, and cooption of regulatory systems to control a different set of genes . We investigated one example in greater detail by measuring the in vitro activity of the S . cerevisiae transcription factor Rpn4p and its orthologs from Candida albicans and Neurospora crassa . Our results suggest that the DNA binding specificity of these proteins has coevolved with the sequences found upstream of the Rpn4p target genes and suggest that Rpn4p has a different function inN . crassa.

Chembiochem, 2004 Dec 3, 5(12), 1647 - 52
Hoechst 33258 selectively inhibits group I intron self-splicing by affecting RNA folding; Disney MD et al.; Fungal pathogens are increasing in prevalence due to an increase in resistant strains and the number of immunocompromised humans . Candida albicans is one of these pathogens, and approximately 40% of strains contain a group I self-splicing intron, which is a potential RNA drug target, in their large subunit rRNA precursor . Here, we report that Hoechst 33258 and derivatives thereof are selective inhibitors of C . albicans group I intron self-splicing with an IC50 of 17 microM in 2 mM Mg2+ . Chemical probing of the intron in the presence of Hoechst 33258 reveals that the folding of several nucleotides in the P4/P6 region of the intron is affected . A nucleotide near the J4/5 region is protected from chemical modification in the presence of Hoechst 33258 and several nearby are more reactive; this suggests that this region is the molecule's binding site . These results expand the available information on small-molecule targeting of RNA and suggest that the RNA-targeting scaffold provided by Hoechst may prove valuable in designing compounds that inhibit the functions of RNA.

Pathol Biol (Paris), 2004 Nov, 52(9), 544 - 9
{Usefulness of panfungal PCR for the diagnosis of fungal infection in immunocompromised patients}; Adam O et al.; The diagnostic of invasive fungal infection is often difficult because of the low sensitivity of fungal culture from infected tissues . Here we have assessed the ability of a panfungal PCR targeted on the DNA region encoding the RNA genes followed by sequencing of the amplification products to detect and identify fungi from tissue biopsies . This assay allowed us to identify the microorganism responsible for an invasive fungal infection in three of our patients . In two cases, hepatosplenic candidiasis was suspected and Candida albicans DNA was detected from liver biopsies . The third patient was cared for a thymome and developed a manubrium osteitis caused by Scedosporium apiospermum.

Structure (Camb), 2004 Nov, 12(11), 1937 - 45
Estriol bound and ligand-free structures of sterol 14alpha-demethylase; Podust LM et al.; Sterol 14alpha-demethylases (CYP51) are essential enzymes in sterol biosynthesis in eukaryotes and drug targets in antifungal therapy . Here, we report CYP51 structures in ligand-free and estriol bound forms . Using estriol as a probe, we determined orientation of the substrate in the active site, elucidated protein contacts with the invariant 3beta-hydroxy group of a sterol, and identified F78 as a key discriminator between 4alpha-methylated and 4alpha,beta-dimethylated substrates . Analysis of CYP51 dynamics revealed that the C helix undergoes helix-coil transition upon binding and dissociation of a ligand . Loss of helical structure of the C helix in the ligand-free form results in an unprecedented opening of the substrate binding site . Upon binding of estriol, the BC loop loses contacts with molecular surface and tends to adopt a closed conformation . A mechanism for azole resistance in the yeast pathogen Candida albicans associated with mutations in the ERG11 gene encoding CYP51 is suggested based on CYP51 protein dynamics.

Folia Microbiol (Praha), 2004, 49(4), 484 - 90
Discrimination between Candida albicans and Candida dubliniensis isolated from HIV-positive patients by using commercial method in comparison with PCR assay; Bujdakova H et al.; Nineteen clinical isolates of Candida albicans and C . dubliniensis were isolated from patients (majority of them HIV-positive) in Slovakia, Brazil, Thailand and Japan . Species discrimination was performed by using growth on CHROMagar Candida, commercial biochemical set API 20C AUX, germ-tube test in human serum, growth at 42 and 45 degrees C on Sabouraud-dextrose agar as well as on CHROMagar Candida, assimilation of D-xylose and methyl alpha-D-glucoside by glass-tube test, and production of chlamydospores . These tests were completed by PCR using Cd-oligo2/F and Cd-oligo2/R primer pair specific for C . dubliniensis . Six clinical isolates were confirmed to be C . dubliniensis, remaining 13 strains were determined as C . albicans . The use of conventional method showed that the determination is markedly influenced by personal evaluation suggesting the necessity of using the combination of many tests to obtain correct results comparing with accurate and rapid PCR assay . For discrimination between C . albicans and C . dubliniensis we recommend the combination of primo-cultivation on CHROMagar, followed by germ-tube test and PCR.

Pediatr Med Chir, 2004 Jan-Feb, 26(1), 57 - 60
{Sepsis by Candida in VLBW neonates: therapy with liposomal Amphotericin B}; Occhipinti E et al.; Candida spp . are recently frequent cause of nosocomial sepsis in neonates admitted in NICU, expecially VLBW . Amphotericin B is used in the treament of infections caused by Candida, but heavy side effects, expecially due to renal toxiticy, prevent often its use in the VLBW neonates . We used a new formulation of Amphotericin (AmBisome--NEXSTAR), in the treatment of 17 VLBW neonates affected by sepsis caused by Candida albicans, admitted in our NICU during two years . Twelve neonates survived, one neonate died for NEC and bowel perforation . The treatment was prolonged for 13 days (7-33 days), the total amount of dose was 65 mg/Kg, none side effect was noted.

J Clin Microbiol, 2004 Nov, 42(11), 4956 - 60
Evaluation of a rapid immunochromatographic assay for identification of Candida albicans and Candida dubliniensis; Marot-Leblond A et al.; Candida dubliniensis was first established as a novel yeast species in 1995 . It is particularly associated with recurrent episodes of oral candidosis in human immunodeficiency virus (HIV)-infected patients, but it has also been detected at other anatomical sites and at a low incidence level in non-HIV-infected patients . It shares so many phenotypic characteristics with C . albicans that it is easily misidentified as such . No rapid, simple, and commercial test that allows differentiation between C . dubliniensis and C . albicans has been developed, until now . Accurate species identification requires the use of genotype-based techniques that are not routinely available in most clinical microbiology diagnostic laboratories . The present study was designed to evaluate the efficiency of a new test (the immunochromatographic membrane {ICM} albi-dubli test; SR2B, Avrille, France) to differentiate between C . albicans and C . dubliniensis . The organisms evaluated were strains whose identities had previously been confirmed by PCR tests and freshly isolated clinical strains and included 58 C . albicans isolates, 60 C . dubliniensis isolates, and 82 isolates belonging to other species of yeast . The ICM albi-dubli test is based on the principle of immunochromatographic analysis and involves the use of two distinct monoclonal antibodies that recognize two unrelated epitopes expressed by both species or specific to only one species . The assay requires no complex instrumentation for analysis and can be recommended for routine use in clinical microbiology laboratories . Results are obtained within 2 h and 30 min and are easy to interpret . This evaluation demonstrated the good performance of this immunochromatographic test for C . albicans and C . dubliniensis isolated on Sabouraud dextrose agar, CHOROMagar Candida, and CandidaSelect, with sensitivities and specificities ranging from 93.1 to 100% . These parameters decreased, however, to 91.4% when the test was performed with yeast isolated with Candida ID.

Int Immunol, 2004 Dec, 16(12), 1761 - 8 Epub 2004 Nov 04.
High dissemination and hepatotoxicity in rats infected with Candida albicans after stress exposure: potential sensitization to liver damage; Correa SG et al.; The liver constitutes the first barrier in the control of hematogenous dissemination for Candida albicans of intestinal origin . The ability of this organ to limit the growth of the yeast and to mount an efficient inflammatory reaction is crucial in determining the outcome of the fungal infection . When rats infected with C . albicans are exposed to chronic varied stress, the cell recruitment is impaired at the site of the infection, the tissue reaction is highly disorganized in target organs and the infection evolution is more severe . At hepatic level, higher fungal burden is associated with hyphal form and the consistent presence of steatosis (fatty liver) . Herein we aimed at characterizing the steatosis associated with C . albicans infection and to provide molecular evidence of the correlation among liver injury markers, stress products and the initiation of the inflammatory tissue reaction . After 3 days of stress and infection, we observed micro and macro steatosis in acinar zone 1 (specific lipid stain), higher lipid peroxidation and increased levels of serum alanine aminotransferase and gamma glutamil transferase . While infection triggered hepatic NO production and arginase activity, stress down-modulated both . Remarkably, defects in levels of TNF-alpha and NO were observed during the first step of the inflammatory response . Our results demonstrate that stress mediators down-regulate the acute inflammatory reaction in the hepatic scenario, promoting a major liver injury with particular immunopathological traits.

Inflammopharmacology, 2004, 12(3), 261 - 70
Cicatrizing and antimicrobial properties of an ozonised oil from sunflower seeds; Rodrigues KL et al.; The ozonised sunflower oil, Bioperoxoil, was tested for its antimicrobial activity against some pathological strains in vitro together with its healing potential against Staphylococcus aureus in vivo . Bioperoxoil was tested against S . aureus, Pseudomonas aeroginosa, Candida albicans, S . typhimurium and Escherichia coli suspensions using the agar diffusion method . Healing experiments were carried out with Wistar rats through topical application of 3.5 mg/ml of the ozonised oil up to the 7th day after inoculation with S . aureus . Bioperoxoil showed anti-inflammatory effects against all strains tested, with MIC values ranging from 2.0 to 3.5 mg/ml . Bioperoxoil also demonstrated protective effects on skin connective tissue and to enhance wound healing during the treatment, as compared to a neomycin-clostebol association used as a positive control . The overall results indicated a significant antimicrobial activity, anti-inflammatory and wound-healing properties for Bioperoxoil, as compared to other antimicrobial agents commercially available.

Inflammation, 2004 Jun, 28(3), 127 - 32
Inflammation induced by inoculation of the joint with Candida albicans; Yordanov M et al.; In humans Candida albicans is the most frequently isolated opportunistic fungal pathogen . In immunocompromized host the balance with the commensal fungus easily turns to life-threatening disseminated infection . The asymptomatic Candida persistence in organs and the recurrent infections suggest continuous circulation of yeast cells and their degradation products . Under certain conditions, joints might become one of the infectious sites . More easily a reactivation and destructive process can be provoked in individuals with established arthritis . We have investigated the joint inflammation caused by inoculation of the paw with live C . albicans, in intact mice and mice with collagen-induced arthritis (CIA) . The results demonstrate that C . albicans infection when localized into the joints caused rapidly progressing septic arthritis . The effect was associated with a strong swelling, a rapid influx of polymorphonuclear (PMN) cells, and an elevated secretion of TNF-alpha and IFN-gamma by lymph node cells . Joint infection exacerbated the established CIA which correlated with an increased level of anti-collagen antibodies.

Ginekol Pol, 2004 Jun, 75(6), 451 - 6
{Occurrence of fungal pathogens in the delivery rooms of a hospital obstetrics department}; Krajewska K et al.; OBJECTIVES: Assessment of the incidence of fungal pathogens in air of the operating rooms from one of the hospitals in Bialystok . MATERIAL AND METHODS: Investigations were conducted in selected rooms of obstetrics department . Material for mycological studies was air sampled at the entrance of hospital building, the entrance to operating room, hall and selected rooms of the department . Fungi were identified using the standard microbial procedures: The monitoring of airborne fungi pollution was done using SAS SUPER 100 (pbi international) . Classification of the isolated fungi was done with an accordance to the current procedures . RESULTS: In the air of 16 rooms of obstetrics department different numbers of fungal colonies from 0 to 560 CFU/1000L of air were isolated . Fungi were not isolated from the air samples of preparing, septic, operating and family deliveries rooms . The highest number of fungi colonies were isolated at the entrance the hospital . The following fungal pathogens isolated from the air were: Candida albicans . non-Candida albicans, Penicillium species, Cladosporium species and Aspergillus species . CONCLUSIONS: 1 . The different number of fungal colonies was found depending on type of the hospital room . 2 . The highest number of fungal colonies was isolated from the air samples of patients rooms 3 . No fungal colonies were isolated from the septic, operating and family deliveries rooms 4 . The main fungal pathogen isolated from the air samples was Candida albicans.

J Biotechnol, 2004 Nov 9, 114(3), 279 - 87
Aspergillus fumigatus CY018, an endophytic fungus in Cynodon dactylon as a versatile producer of new and bioactive metabolites; Liu JY et al.; Aspergillus fumigatus CY018 was recognized as an endophytic fungus for the first time in the leaf of Cynodon dactylon . By bioassay-guided fractionation, the EtOAc extract of a solid-matrix steady culture of this fungus afforded two new metabolites, named asperfumoid (1) and asperfumin (2), together with six known bioactive compounds including monomethylsulochrin, fumigaclavine C, fumitremorgin C, physcion, helvolic acid and 5alpha,8alpha-epidioxy-ergosta-6,22-diene-3beta-ol as well as other four known compounds ergosta-4,22-diene-3beta-ol, ergosterol, cyclo(Ala-Leu) and cyclo(Ala-Ile) . Through detailed spectroscopic analyses including HRESI-MS, homo- and hetero-nuclear correlation NMR experiments (HMQC, COSY, NOESY and HMBC), the structures of asperfumoid and asperfumin were established to be spiro-(3-hydroxyl-2,6-dimethoxyl-2,5-diene-4-cyclohexone-(1,3')-5'-methoxyl-7'-methyl-(1'H, 2'H, 4'H)-quinoline-2',4'-dione) and 5-hydroxyl-2-(6-hydroxyl-2-methoxyl-4-methylbenzoyl)-3,6-dimethoxyl-benzoic methyl ester, respectively . All of the 12 isolates were subjected to in vitro bioactive assays against three human pathogenic fungi Candida albicans, Tricophyton rubrum and Aspergillus niger . As a result, asperfumoid, fumigaclavine C, fumitremorgin C, physcion and helvolic acid were shown to inhibit C . albicans with MICs of 75.0, 31.5, 62.5, 125.0 and 31.5 microg/mL, respectively.

Indian J Physiol Pharmacol, 2004 Apr, 48(2), 225 - 9
Non-secretor status; a predisposing factor for vaginal candidiasis; Kulkarni DG et al.; A secretor is an individual who secretes blood group antigens into body fluids such as saliva, sweat, tears, semen and serum . An attempt has been made to establish the correlation between the secretor status and susceptibility to vaginal candidiasis . The secretor status was determined by haemagglutination inhibition technique . The presence of Candida albicans infection was detected by direct microscopy of the wet smear and confirmed by germ tube test and corn meal agar test . It was observed that out of the 64 patients, 15 were secretors and 49 were non-secretors . However 43 subjects were secretors and 13 non-secretors among the 56 controls . Thus prevalence of vaginal candidiasis was significantly higher in non-secretor group (P<0.01) . The absence of blood group antigens in the body fluids and the lack of enzyme glycosyl transferase enhance the attachment of yeast to the epithelial cell and render the non-secretor more prone to infection.

J Biol Chem, 2005 Jan 14, 280(2), 1051 - 60 Epub 2004 Nov 01.
Mnt1p and Mnt2p of Candida albicans Are Partially Redundant {alpha}-1,2-Mannosyltransferases That Participate in O-Linked Mannosylation and Are Required for Adhesion and Virulence; Munro CA et al.; The MNT1 gene of the human fungal pathogen Candida albicans is involved in O-glycosylation of cell wall and secreted proteins and is important for adherence of C . albicans to host surfaces and for virulence . Here we describe the molecular analysis of CaMNT2, a second member of the MNT1-like gene family in C . albicans . Mnt2p also functions in O-glycosylation . Mnt1p and Mnt2p encode partially redundant alpha-1,2-mannosyltransferases that catalyze the addition of the second and third mannose residues in an O-linked mannose pentamer . Deletion of both copies of MNT1 and MNT2 resulted in reduction in the level of in vitro mannosyltransferase activity and truncation of O-mannan . Both the mnt2Delta and mnt1Delta single mutants were significantly reduced in adherence to human buccal epithelial cells and Matrigel-coated surfaces, indicating a role for O-glycosylated cell wall proteins or O-mannan itself in adhesion to host surfaces . The double mnt1Deltamnt2Delta mutant formed aggregates of cells that appeared to be the result of abnormal cell separation . The double mutant was attenuated in virulence, underlining the importance of O-glycosylation in pathogenesis of C . albicans infections.

Mycopathologia, 2004 Aug, 158(2), 181 - 6
Onychomycosis in Cali, Colombia; Alvarez MI et al.; This study presents the epidemiological and mycological aspects of 299 patients with nail lesions who were referred to three diagnostic laboratories in the city of Cali . The diagnosis of mycoses was established through visualization of mycotic structures in a direct microscopic examination of skin scrapings and by isolation . Onychomycosis was found in 183 cases (61.2%), of which 141 were in toenails (44 in males and 97 in females), 38 in fingernails (9 males and 29 females), and 4 cases in toenails and fingernails simultaneously (all females) . No statistically significant relation was found between sex and onychomycosis . Yeasts accounted for 40.7% of the mycoses, dermatophytes for 38%, nondermatophyte molds for 14% and the etiology was mixed in the remaining cases (7.3%) . Candida albicans was the most commonly isolated yeast species; the most common dermatophyte was Trichophyton rubrum and Fusarium spp . and Scytalidium dimidiatum were the most common nondermatophytic molds . Them common fungi found in fingernails were yeasts; in toenails dermatophytes were more prevalent (chi2 with Yates' correction = 19.75, P= 0.000088) . Yeasts were observed more frequently in females while dermatophytes were more common in males . The difference between these two etiologic groups was statistically significant (chi2 with Yates' correction = 7.43, P = 0.0064); no relation was observed according to age.

Eur J Appl Physiol . 2004 Oct 29; {Epub ahead of print}
Norepinephrine as mediator in the stimulation of phagocytosis induced by moderate exercise; Ortega E et al.; During intensive exercise the stimulation of phagocytosis is mediated by "stress hormones" . During moderate exercise, however, such mediation is less clear . The influence of moderate exercise (45 min at 55% maximal oxygen uptake) on the phagocytic capacity of neutrophils was evaluated in sedentary men . The exercise stimulated phagocytosis of Candida albicans, and the stimulation was maintained for at least 24 h . The possible neuroendocrine mediators were then investigated . Stimulation of phagocytosis was found after incubating neutrophils from sedentary individuals, who were in a basal state, with plasma from exercised individuals . Immediately after exercise, there was a significant increase in the concentration of norepinephrine, but not of epinephrine or cortisol . Incubation of neutrophils with this post-exercise physiological concentration of norepinephrine also stimulated phagocytosis, and the effect was blocked by both propranolol and phentolamine . The norepinephrine-augmented phagocytosis was accompanied by an increase in intracellular levels of cAMP, but not of cGMP or calcium . In conclusion, moderate exercise performed by sedentary people stimulates the phagocytic capacity of neutrophils, and the stimulation lasts for at least 24 h . Norepinephrine mediates the stimulation, although other mechanisms could be involved during the recovery period.

Bone Marrow Transplant, 2004 Nov, 34(10), 891 - 5
Candidaemia in allogeneic stem cell transplant recipients: low risk without fluconazole prophylaxis; Jantunen E et al.; Invasive fungal infections (IFI) are common in allogeneic SCT recipients . We have reviewed our experience of IFI with special reference to candidaemia in 685 adult patients transplanted in 1983-2002 . The donor was a matched sibling in 505 patients and an unrelated donor in 180 patients . A BM graft was used in 561 patients and a PB graft in 124 patients . Fluconazole prophylaxis was not used during the study period . Definite or probable IFI was observed in 60 patients (8.7%) with a dominance of Aspergillus infections (46 patients, incidence 6.7%) . Candidaemia was found only in nine patients (1.3%) . The causative agents were Candida albicans (n=8), C . krusei (n=2), and C . glabrata (n=1); in two patients, two causative agents were found . The median time to the diagnosis of candidaemia was 53 days (range 6-249 days) post transplant . Seven patients were neutropaenic at diagnosis, and four patients had experienced acute GVHD . All patients received antifungal therapy, but only one patient was cured . According to this study, candidaemia was a rare event in allogeneic SCT recipients . Thus, systematic prophylaxis against Candida infections might not be indicated . The prognosis of established infections is still poor due to comorbid conditions, notably GVHD.

Chem Pharm Bull (Tokyo), 2004 Nov, 52(11), 1353 - 5
A new steroidal saponin from Dioscorea cayenensis; Sautour M et al.; The new 26-O-beta-D-glucopyranosyl-3beta,26-dihydroxy-20,22-seco-25(R)-furost-5-en-20,22-dione-3-O-alpha-L-rhamnopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->4)-{alpha-L-rhamnopyranosyl-(1-->2)}-beta-D-glucopyranoside (1), along with the known methyl protodioscin (2), asperoside (3) and prosapogenin A of dioscin (4) were isolated from the rhizomes of Dioscorea cayenensis LAM.-HOLL (Dioscoreaceae) . Their structures were established mainly on the basis of 600 MHz 2D-NMR spectral data . 4 exhibited antifungal activity against the human pathogenic yeasts Candida albicans, C . glabrata and C . tropicalis (MICs of 20.8, 6.25, 25 microg/ml, respectively), whereas saponins 1-3 were inactive.

Przegl Lek, 2004, 61(5), 467 - 72
{Clinical and mycological examinations of oral mucosa in cyclosporine A treated patients after renal transplantation}; Tyrzyk S et al.; Cyclosporine A (CsA) is one of the basic and often used immunosupressive agents . It is used to prevent rejection of allogenic transplants and to cure many diseases with autoimmunological components . The development of transplantology and frequent presence of autoaggressive disorders causes increased use of CsA, which intensifies problem of side-effects . Gingival overgrowth, oral mucosa pathologies related with bacterial, viral and fungal infections and neoplasma are described as most frequent side-effects of cyclosporine A in the oral cavity . The aim of this study was to assess the frequency of presence of different types of oral mucosa lesions in cyclosporine A-treated patients after renal transplantation . Thirty renal transplant recipients aged 11-64 years (including 13 females and 17 males) with cyclosporine treatment period from 6 months to 10 years were examined . Pathological changes on the oral mucosa were noted in 18 patients (60%) . The fungal infections were the most often observed pathology (46.7% of examined persons) . In mycological tests Candida albicans was most often isolated . Other species: C . kefyr, C . parapsilosis and C . tropicalis were also found . In 4 cases cheilitis angularis was additionally diagnosed . In 16.7% of CsA-treated patients, pathological changes of epithelium with homogenic leucoplakia features were observed . Hypertrophic lesions of oral mucosa in 10% of cases and many small erosions of inflammated mucosa of hard palate in 1 case were also noted . Our observations show connection between incidence of oral mucosa pathologies and immunosupressive treatment . This suggests that interdisciplinary cooperation is necessary for early start of prevention and treatment program.

J Lab Clin Med, 2004 Oct, 144(4), 208 - 14
Fungal susceptibility to zinc deprivation; Lulloff SJ et al.; Calprotectin is a neutrophil-derived antimicrobial protein that competes with microorganisms for zinc . The zinc-specific effect of calprotectin against Candida albicans appears to be related to this organism's marked susceptibility to deprivation of this metal . However, it is not known whether this susceptibility is particular to C albicans or whether it is a characteristic of pathogenic fungi in general . As a means of deciding between these 2 possibilities, we undertook the study reported here to compare the susceptibility to zinc deprivation of 6 other pathogenic fungal species in addition to that of C albicans . We tested the effect of metals in reversing growth inhibition of the 7 fungi against abscess-fluid supernatant (a source of calprotectin) and 3 chemical chelators . Data were expressed as the concentration of metal required to bring about 50% restoration of growth . Zinc was found to be much more potent than the other metals tested in reversing growth inhibition of all the organisms by human abscess fluid and all 3 chemical chelators . Copper and manganese also had some effect . In some cases, chelator stability constants were higher for other metals than for zinc; in particular, although diethylenetriaminopentaacetic acid has a stability constant for iron almost 10(10) times greater than that for zinc, zinc was more effective than iron in reversing growth inhibition by this chelator against all of the organisms . These results suggest that marked susceptibility to zinc deprivation is a general characteristic of pathogenic fungi.

J Antimicrob Chemother, 2004 Dec, 54(6), 1096 - 102 Epub 2004 Oct 27.
Alternative dosing regimens of liposomal amphotericin B (AmBisome) effective in treating murine systemic candidiasis; Adler-Moore JP et al.; OBJECTIVES: This study was done to determine whether high dose AmBisome (4-20 mg/kg), given intermittently, could reduce the frequency of dosing needed to treat murine systemic candidiasis when compared with conventional daily treatment . METHODS: Mice were immunosuppressed with cyclophosphamide every 3 days, beginning day -3 before challenge with log(10) 5.0 cfu Candida albicans . Treatment was begun 48-72 h post-challenge with daily or intermittent dose regimens of AmBisome, followed by determination of kidney cfu for up to 1 month post-treatment . RESULTS: A single AmBisome dose of 4 mg/kg was as effective as four daily, 1 mg/kg treatments . A total of 8 mg/kg, given as 4 mg/kg on days 2 and 4, or as 5 mg/kg on day 2 followed by 1 mg/kg on days 3, 4, and 5, also produced comparable efficacy . While 20 mg/kg given day 2, 4 and 6 post-challenge as a 1 week loading dose, followed by one 10 mg/kg treatment on day 13, decreased the fungal burden by up to 5 logs compared with controls (log(10) 2.3 cfu/g and log(10) 7.5 cfu/g, respectively), 20 mg/kg given Monday, Wednesday and Friday for 5 weeks, reduced the fungal burden to undetectable levels (i.e . log(10) 1.0 cfu) . CONCLUSIONS: Significant reduction or clearance of kidney cfu, following intermittent, high dose AmBisome treatment, indicated that non-daily dosing regimens could be successfully used instead of conventional daily dosing to treat established C . albicans infection in immunosuppressed mice.

Jpn J Infect Dis, 2004 Oct, 57(5), S9 - 10
Murine model of Kawasaki disease induced by mannoprotein-Beta-glucan complex, CAWS, obtained from Candida albicans; Ohno N; Intraperitoneal administration of CAWS (water-soluble extracellular polysaccharide fraction obtained from the culture supernatant of Candida albicans) to mice induces coronary arteritis similar to Kawasaki disease . We analyzed differences in the production of cytokines involved in the occurrence of coronary arteritis among mouse strains, C3H/HeN, C57BL/6, DBA/2 and CBA/J . The incidence of arteritis was 100% in C57BL/6, C3H/HeN and DBA/2 mice, but only 10% in CBA/J mice . The coronary arteritis observed in DBA/2 mice was the most serious, with several mice expiring during the observation period . The CAWS-sensitive strains revealed increased levels of IL-6 and IFN-gamma during the course of a specific response to CAWS by spleen cells . In contrast, IL-10 levels were observed to increase markedly in CAWS-resistant CBA/J mice, but not the CAWS-sensitive strains . However, TNF-alpha levels were more elevated only in DBA/2 mice . The difference in disease development and cytokine production strongly suggests that the genetic background of the immune response to CAWS contributes to the occurrence of coronary arteritis.

Jpn J Infect Dis . 2004 Oct;57(5):S15.
In vivo role of myeloperoxidase for the host defense; Aratani Y et al.; Myeloperoxidase (MPO) is located within neutrophils capable of producing HOCl . To define the in vivo role of MPO, we have generated MPO-knockout (MPO-KO) mice . The mice without MPO developed normally . However, MPO-KO mice showed severely reduced cytotoxicity to various microorganisms such as Candida albicans, Aspergillus fumigatus, and Klebsiella pneumoniae, demonstrating that MPO-dependent oxidative system is important for host defense against fungi and bacteria, although the effect varies from species to species of pathogens . To compare the importance of MPO and NADPH-oxidase for host defense, MPO-KO and chronic granulomatous disease (CGD) mice were infected with different doses of C . albicans, and their infection severity was analyzed . CGD mice exhibited increased mortality and tissue fungal burden in a dose-dependent manner, whereas normal mice showed no symptoms . Interestingly, at the highest dose, the mortality of MPO-KO mice was comparable to CGD mice, but was the same as normal mice at the lowest dose . These results suggest that MPO and NADPH-oxidase are equally important for early host defense against a large inocula of Candida.

Antimicrob Agents Chemother, 2004 Nov, 48(11), 4450 - 2
Antimicrobial and antileishmanial activities of hypocrellins A and B; Ma G et al.; Hypocrellins A and B were evaluated for in vitro antimicrobial and antileishmanial activities . Hypocrellin A exhibited promising activity against Candida albicans and moderate activity against Staphylococcus aureus, methicillin-resistant S . aureus, Pseudomonas aeruginosa, and Mycobacterium intracellulare . Hypocrellin B showed weak antimicrobial activities . Hypocrellin A exhibited potent antileishmanial activity, while hypocrellin B was only moderately active . These results of promising antifungal and antileishmanial activity of hypocrellin A may be useful for further structure-activity relationship and in vivo studies.

Antimicrob Agents Chemother, 2004 Nov, 48(11), 4377 - 86
Molecular mechanisms of primary resistance to flucytosine in Candida albicans; Hope WW et al.; Primary resistance in Candida albicans to flucytosine (5-FC) was investigated in 25 strains by identifying and sequencing the genes FCA1, FUR1, FCY21, and FCY22, which code for cytosine deaminase, uracil phosphoribosyltransferase (UPRT), and two purine-cytosine permeases, respectively . These proteins are involved in pyrimidine salvage and 5-FC metabolism . An association between a polymorphic nucleotide and resistance to 5-FC was found within FUR1 where the substitution of cytidylate for thymidylate at nucleotide position 301 results in the replacement of arginine with cysteine at amino acid position 101 in UPRT . Isolates that are homozygous for this mutation display increased levels of resistance to 5-FC, whereas heterozygous isolates have reduced susceptibility . Three-dimensional protein modeling of UPRT suggests that the Arg101Cys mutation disturbs the quaternary structure of the enzyme, which is postulated to compromise optimal enzyme activity . A single resistant isolate, lacking the above polymorphism in FUR1, has a homozygous polymorphism in FCA1 that results in a glycine-to-aspartate substitution at position 28 in cytosine deaminase.

Antimicrob Agents Chemother, 2004 Nov, 48(11), 4337 - 41
Inhibition of adherence and killing of Candida albicans with a 23-Mer peptide (Fn/23) with dual antifungal properties; Klotz SA et al.; Candida albicans adheres to host tissue and then proliferates in order to establish a commensal as well as a pathogenic state . Specific adherence to proteins is provided by several surface adhesins of Candida . Two well-studied proteins, Als1p and Als5p, do not require energy for adherence to occur (dead as well as living cells adhere) and have a multiplier effect of cell-cell aggregation that mediates the formation of microcolonies of Candida cells . The entire process is spontaneous, reversible, and stable for physiologically relevant chemical and physical forces . This adherence process is inhibited by the addition of free peptide ligands, including a 23-mer derived from fibronectin (Fn/23) that binds to the adhesins through H bond formation . Adherence was measured by determining the number of yeast cells that adhered to 90-microm-diameter polyethylene glycol (PEG) beads with a 7-mer peptide (KLRIPSV) synthesized on the surfaces of the beads . The concentration of the Fn/23 peptide that inhibited the adherence of cells to the peptide-coated beads by 50% was 4 to 5 microM, and the magnitudes of adherence were similar regardless of the presence or absence of physiologic salt concentrations . The minimum fungicidal concentration of Fn/23 was 2 to 4 microM in water, but there was no killing in physiologic salt concentrations . Peptides from the C and N termini or the center sequence of Fn/23 had no effect on inhibition of adherence and little effect on fungal viability . The fungicidal effect was similar to that seen with 23-, 19-, and 18-mer peptides derived from porcine myeloid cells, a Helicobacter pylori ribosomal protein, and a hybrid of cecropin and magainin, respectively . However, these fungicidal peptides did not inhibit C . albicans adherence to the peptide-coated PEG beads . This dual property of Fn/23, i.e., inhibition of adherence and killing of C . albicans, may provide important adjuvant effects in the treatment of disease caused by this fungus.

Mycoses, 2004 Oct, 47(9-10), 450 - 3
Lethal otogenic Candida meningitis; Koch S et al.; History revealed a chronic obstructive pulmonary condition which had been treated with prednisolone for a long time . There was a raised temperature with further signs of an acute inflammatory underlying disease and internal hydrocephalus . After performing trepanation, the symptoms of raised intercerebral pressure ceased . Candida albicans could be detected microbiologically in the cerebrospinal fluid . There was no pneumonia at the time of admission . Despite instituting immediate intensive care with administration of antibiotics and antimycotics, the patient died 11 days after inpatient admission . Autopsy revealed a C . albicans mycosis originating from the right middle ear with extensive suppurative meningitis, which was the immediate cause of death . Confluent bronchopneumonia had developed in both lower lung lobes at the time of death, but did not show any signs of mycosis and had contributed indirectly to the death of the patient.

Mycoses, 2004 Oct, 47(9-10), 435 - 41
Oral candidosis and associated Candida species in HIV-infected Cambodians exposed to antimycotics; Schmidt-Westhausen AM et al.; Although human immundeficiency virus (HIV) infection is endemic in Southeast Asia, data on oral mycotic flora in this disease in Asians are sparse . The aim of this study was to determine the prevalence of Candida species in HIV-infected Cambodians with oral candidosis, unexposed (group 1) and exposed to antimycotics (group 2) and a healthy population (group 3) . In 161 HIV patients with oral candidosis (group 1: 121 pts; group 2: 40 pts) and in 81 controls (group 3) swab samples of tongue and palate were obtained . Oral candidosis was detected in 100 and 70% of groups 1 and 2 respectively . Candida spp . were isolated from 91 and 100% of groups 1 and 2, respectively, and from 79% of controls . Candida albicans was the most common, with non-albicans species such as C . tropicalis and C . krusei being notable . Our data indicate that variants of oral candidal infections in HIV disease are similar to those seen in the pre-HAART era . The particularly high rate of C . krusei isolation in all groups is noteworthy.

Mycoses, 2004 Oct, 47(9-10), 422 - 7
Clinical and mycological efficacy of single-day oral treatment with itraconazole (400 mg) in acute vulvovaginal candidosis; Urunsak M et al.; This study aimed to investigate the effectiveness of single-day oral treatment with itraconazole in acute vulvovaginal candidosis (VVC) . Vaginitis was demonstrated by both detection of yeast cells and pseudohyphae formation on microscopic examination of vaginal discharge and mycological culture as well as by the clinical signs and symptoms . Clinical and mycological examinations of the 52 patients were performed before, 1 week (short-term) and 4 weeks (long-term) after single-day oral treatment with itraconazole 200 mg b.i.d . The causative yeast fungi were: Candida albicans (76.9%), C . glabrata (9.6%), C . kefyr (9.6%) and C . krusei (3.9%), respectively . In short- and long-term examinations, clinical cure rates were found to be 61.5% and 90.4%, and mycological cure rates were 63.5% and 90.4%, respectively . Itraconazole was found to be 95.0% effective with C . albicans and 75.0% with other Candida species . It is concluded that treatment of acute VVC with itraconazole is safe and effective in the long-term.

Mycoses, 2004 Oct, 47(9-10), 402 - 6
In vitro susceptibility testing of amorolfine in pathogenic fungi isolated from dermatomycosis patients in China; Li RY et al.; The antifungal susceptibility of isolates from Chinese dermatomycosis patients to amorolfine was investigated following National Committee for Clinical Laboratory Standards (NCCLS) protocols . In total, 383 isolates were tested, including 132 strains from tinea pedis, 148 strains from tinea corporis/cruris, and 103 strains from onychomycosis . The minimum inhibitory concentration (MIC) of amorolfine against dermatophytes ranged from 0.01 to 0.08 microg ml(-1) . The MIC(50) and MIC(90) of amorolfine for Trichophyton rubrum were both equal to 0.04 micro ml(-1); for T . mentagrophytes these MICs were 0.04 microg ml(-1) and 0.08 microg ml(-1) respectively; and for Epidermophyton floccosum they were 0.02 microg ml(-1) and 0.04 microg ml(-1) respectively . The MIC range of amorolfine against Candida parapsilosis was 0.5-16 microg ml(-1) . MIC(50) and MIC(90) for C . parapsilosis were 0.5 and 2 microg ml(-1) . MIC ranges of amorolfine against Scopulariopsis spp . and Acremonium spp . were 0.5-4 and 2-8 microg ml(-1), respectively . Candida albicans, Fusarium solani and Aspergillus flavus required relatively higher concentrations of amorolfine to inhibit their growth (MIC 0.125-64 microg ml(-1), MIC(50) and MIC(90) were 4 and 64 microg ml(-1)) . The results demonstrated that amorolfine is the only topical agent that has such a potent antifungal activity and a broad spectrum against a wide range of pathogenic fungi.

Mycoses, 2004 Oct, 47(9-10), 397 - 401
Comparative evaluation of Sensititre YeastOne vs . the NCCLS M27A protocol and E-test for antifungal susceptibility testing of yeasts; Lombardi G et al.; A recently developed microdilution method (Sensititre) YeastOne) may represent a valid alternative to the National Committee for Clinical Laboratory Standards (NCCLS) method for routine testing . The Medical Mycology Committee of the Associazione Microbiologi Clinici Italiani (AMCLI) decided to evaluate its reproducibility and reliability compared with the NCCLS M27A protocol and the E-test . Nineteen strains each of Candida albicans and Ca . parapsilosis, isolated from systemic infections, were tested against amphotericin B, flucytosine, ketoconazole, itraconazole, and fluconazole . All the participating laboratories tested the YeastOne panels, while the E-test and the NCCLS method were performed by two laboratories each . Interlaboratory reproducibility showed a good correlation (from 95% for amphotericin B to 92.5% for flucytosine) . The agreement between NCCLS and YeastOne ranged from 95 (ketoconazole and itraconazole) to 100% (amphotericin B and flucytosine), whereas the agreement between E-test and YeastOne ranged from 72.5 (fluconazole) to 100% (amphotericin B and flucytosine) . The Sensititre YeastOne panels appear to be an excellent alternative to both the E-test and the NCCLS protocol for antifungal susceptibility testing.

Microbiol Immunol, 2004, 48(10), 783 - 5
Acetate-mediated production of orotic acid by ura3 mutants of Candida albicans; Nezu T et al.; Candida albicans ura3 mutants were found to produce large amounts of orotic acid when the growth medium was supplemented with sodium acetate . Experiments with 13C-labeled acetate showed that the acetate served as a precursor of orotic acid . This system of acetate-mediated production of orotic acid is similar to other documented microbial producers in yield but unique for its acetate requirement.

Infect Immun, 2004 Nov, 72(11), 6633 - 41
The hyphal and yeast forms of Candida albicans bind the complement regulator C4b-binding protein; Meri T et al.; Candida albicans, an important pathogenic yeast, activates all three pathways of the complement system . To understand how this yeast evades the effects of the activated system, we have analyzed the binding of the classical pathway inhibitor C4b-binding protein (C4BP) by C . albicans . Purified native as well as recombinant C4BP bound dose dependently to the yeast and hyphal forms, as shown by multiple methods, such as confocal microscopy, flow cytometry, a novel enzyme-linked immunosorbent assay, absorption from human serum, and direct binding assays with purified proteins . A prominent binding site was identified at the tip of the germ tube, a structure that is considered important for tissue penetration and pathogenesis . The binding site in C4BP was localized to the two N-terminal complement control protein domains by using recombinant deletion constructs and site-specific monoclonal antibodies . As the alternative pathway inhibitors factor H and FHL-1 also bind to C . albicans, the binding of all three plasma proteins was compared . Simultaneous binding of the classical regulator C4BP and the alternative pathway regulator factor H was demonstrated by confocal microscopy . In addition, FHL-1 competed for binding with C4BP, suggesting that these two related complement regulators bind to the same structures on the yeast surface . The surface-attached C4BP maintains its complement regulatory activities and inactivates C4b . The surface-attached human C4BP serves multiple functions relevant for immune evasion and likely pathogenicity . It inhibits complement activation at the yeast surface and, in addition, mediates adhesion of C . albicans to host endothelial cells.

Infect Immun, 2004 Nov, 72(11), 6230 - 6
Cell wall mannan and cell surface hydrophobicity in Candida albicans serotype A and B strains; Masuoka J et al.; Cell surface hydrophobicity contributes to the pathogenesis of the opportunistic fungal pathogen Candida albicans . Previous work demonstrated a correlation between hydrophobicity status and changes in the acid-labile, phosphodiester-linked beta-1,2-oligomannoside components of the N-linked glycans of cell wall mannoprotein . Glycan composition also defines the two major serotypes, A and B, of C . albicans strains . Here, we show that the cell surface hydrophobicity of the two serotypes is qualitatively different, suggesting that the serotypes may differ in how they modulate cell surface hydrophobicity status . The cell wall mannoproteins from hydrophilic and hydrophobic cells of both serotypes were compared to determine whether the glycan differences due to serotype affect the glycan differences due to hydrophobicity status . Composition analysis showed that the protein, hexose, and phosphate contents of the mannoprotein fraction did not differ significantly among the strains tested . Electrophoretic profiles of the acid-labile mannan differed only with hydrophobicity status, not serotype, though some strain-specific differences were observed . Furthermore, a newly available beta-1,2-oligomannoside ladder allowed unambiguous identification of acid-labile mannan components . Finally, to assess whether the acid-stable mannan also affects cell surface hydrophobicity status, this fraction was fragmented into its component branches by acetolysis . The electrophoretic profiles of the acid-stable branches were very similar regardless of hydrophobicity status . However, differences were observed between serotypes . These results support and extend our current model that modification of the acid-labile beta-1,2-oligomannoside chain length but not modification of the acid-stable region is one common mechanism by which switching of cell surface hydrophobicity status of C . albicans strains occurs.

Infect Immun, 2004 Nov, 72(11), 6206 - 10
Regulation of Candida albicans morphogenesis by fatty acid metabolites; Noverr MC et al.; Candida albicans is an opportunistic dimorphic fungus that inhabits various host mucosal sites . Conversion from the yeast to the hyphal form has been associated with increased virulence and mucosal invasiveness . C . albicans morphogenesis is regulated by multiple signals and signaling pathways . However, signals that control morphogenesis in vivo are unknown . We investigated the effects of host long chain fatty acids, eicosanoids, and bacterial short chain fatty acids on control of germination . None of the C18 or C20 fatty acids tested had an effect on enhancing germ tube formation (arachidonic acid, oleic acid, linolenic acid, or gamma-linolenic acid) . Among the different eicosanoids, both prostaglandin E2 and thromboxane B2 significantly enhanced serum-induced germination by C . albicans . Addition of antiprostaglandin or antithromboxane antibodies to serum alone inhibited germ tube formation by almost 30%, while control antibody had no effect, indicating that these eicosanoids are major morphogenic factors in the serum . Since these molecules also bind to albumin, this may also explain the hyphal transforming activity in serum that associates with albumin . Interestingly, short chain fatty acids (butyric acid), the product of lactic acid bacteria (LAB), inhibited germination . In addition, LAB culture supernatants as well as live LAB also inhibited C . albicans morphogenesis . Overall, these results indicate that fatty acid metabolites and fatty acid pathways can up-regulate and down-regulate germination in C . albicans.

J Hosp Infect, 2004 Nov, 58(3), 200 - 3
Genotyping analysis of colonizing candidal isolates from very-low-birthweight infants in a neonatal intensive care unit; Huang YC et al.; To analyse the relatedness of colonizing candidal isolates from very-low-birthweight infants hospitalized in a neonatal intensive care unit (NICU), we prospectively collected 86 candidal isolates from 20 infants, including 67 isolates of Candida albicans from 15 infants, 17 isolates of Candida parapsilosis from five infants and two isolates of Candida glabrata from one infant, who also had C . albicans colonization, over a one-year period . All 86 isolates were genotyped by infrequent-restriction-site polymerase chain reaction (IRS-PCR) and electrophoretic karyotyping (EK) with pulsed-field gel electrophoresis . A total of 15 genotypes were identified by IRS-PCR and 12 genotypes by EK . Some infants shared a common genotype . From a single infant, an identical genotype was found in 11 of 13 cases where at least two isolates of same Candida species were available for genotyping analysis, regardless of anatomical site, how many isolates were recovered or how many times . Should an infant harbour a candidal strain, they may harbour this strain at multiple sites and for a prolonged period.

Bioorg Med Chem Lett, 2004 Dec 6, 14(23), 5831 - 3
Screening of antimicrobial activity of diarylamines in the 2,3,5-trimethylbenzo{b}thiophene series: a structure-activity evaluation study; Ferreira IC et al.; Gram positive (Bacillus cereus, B . subtilis), Gram negative (Pseudomonas aeruginosa, Escherichia coli) bacteria, and Candida albicans as a representative of fungi were used for screening the in vitro antimicrobial activity of diarylamines in the 2,3,5-trimethylbenzo{b}thiophene series bearing different substituents, synthesized by us using the palladium-catalyzed C-N coupling methodology . The minimal inhibitory concentration (MIC) and structure-activity relationships (SARs) were evaluated.

Bioorg Med Chem, 2004 Nov 15, 12(22), 5881 - 9
Stereoselective synthesis and antifungal activity of (Z)-trans-3-azolyl-2-methylchromanone oxime ethers; Emami S et al.; A series of (Z)-trans-3-azolyl-2-methylchromanone oxime ethers were stereoselectively synthesized and tested for in vitro antifungal activity . Many of these derivatives exhibit high activity against Candida albicans, Saccharomyces cerevisiae, Aspergillus niger, and Microsporum gypseum.

J Nat Prod, 2004 Oct, 67(10), 1732 - 5
A novel antimicrobial indolizinium alkaloid from Aniba panurensis; Klausmeyer P et al.; Activity-guided fractionation of an Aniba panurensis organic solvent extract has led to the isolation of the novel alkaloid 6,8-didec-(1Z)-enyl-5,7-dimethyl-2,3-dihydro-1H-indolizinium, as the trifluoroacetic acid salt (1) . Its structure was determined by NMR and mass spectrometry . Bioassays performed in vitro demonstrated toxicity of compound 1 to a drug-resistant strain of Candida albicans.

Acta Pol Pharm, 2004 Mar-Apr, 61(2), 127 - 33
Chiral mixed ligand Co(II) and Ni(II) complexes: synthesis and biological activity; Shivankar VS et al.; Chiral mixed ligand (CML) transition metal complexes of the type MQL.2H2O, where M is Co(II)/Ni(II), Q is deprotonated 8-hydroxyquinoline and L is a deprotonated chiral saccharide such as (+)-glucose and (-)-fructose, have been synthesized . The metal complexes have been characterized on the basis of elemental analysis and various physicochemical techniques such as molar conductance, specific rotation measurements, magnetic, spectral and thermal studies . The cup-plate method has been used to study the antibacterial activity of the compounds against some of the pathogenic bacteria such as C . diphtheriae, E . coli, S . typhi, S . dysenteriae, S . aureus and V . cholerae . The antifungal activity of the complexes against some of the pathogenic fungi such as Candida albicans and Aspergillus niger has been studied by the tube dilution method . The results have been compared against those of controls, which were screened simultaneously . The complexes have been screened for acute oral toxicity in albino rats . The method of Litchfield and Wilcoxon has been used to determine the LD50 values.

Lasers Surg Med, 2004, 35(4), 259 - 62
Fungicidal effect of diode laser irradiation in patients with denture stomatitis; Maver-Biscanin M et al.; BACKGROUND AND OBJECTIVE: Denture stomatitis (DS) is a common inflammatory condition that affects denture wearers . The aim of this study was to examine, in vivo, the effect of diode laser irradiation on fungal growth in both the palatal mucosa and in denture base materials, in denture wearing patients . STUDY DESIGN/MATERIALS AND METHODS: In total, 70 patients with clinical study design evidence of DS participated in this parallel, single blind, and placebo controlled study . The subjects were randomly assigned to one of four different treatment regimens: (1) irradiation with a 685 nm wavelength laser for 10 minutes (30 mW); (2) irradiation with a 830 nm wavelength laser for 5 minutes (60 mW) . A semiconductor diode laser, BTL-2000 (BTL-2 Dravotnicka Technika, Prague, Czech Republic), was used in both treatment cases using an energy density of 3.0 J/cm(2) and a continuous working mode for five consecutive days; (3) placebo-sham irradiation of patients; (4) antimicotic-self treatment of patient's palatal mucosa with an antifungal oral gel and the use of an antiseptic solution for their dentures . The effect of laser light on fungal growth in vivo was evaluated after final treatment using the swab method and a semi-quantitative estimation of Candida albicans colonies cultivated on agar plates . RESULTS: A fungicidal effect was achieved in the laser treated and antimicotic treated groups, whereas most subjects in the placebo group were found to have unchanged conditions on both their palate (P = 0,004) and dentures (P < 0,001) . CONCLUSIONS: Light from a low-power laser (LLLT) may be valuable in the treatment of DS . This is of great importance since the rate of recurrence of disease is high, whereas an optimal treatment modality has not yet been found . (c) 2004 Wiley-Liss, Inc.

Rev Alerg Mex, 2004 Jul-Aug, 51(4), 145 - 50
{Most common allergens in allergic patients admitted into a third-level hospital}; Ortega EV et al.; BACKGROUND: Since year 3000 BC, pollenology (science that studies allergens) was already known . In the year 49 BC, Herodotus recognized the first case of hay fever, Jacob Constant (17th century) described the first case of allergic rhinitis and related it to "something" (allergens) that roses emitted . Charles Backley (1873) proved the ability of the skin to determine allergic reactions to allergens . Today, it is well-known that allergens are able to react with environmental exposure, plants and pollens . OBJECTIVE: To know the most common allergens in allergic patients who are admitted into a tertiary care hospital . MATERIAL AND METHODS: 3,172 electronic expedients of registered allergic patients were examined . Only 356 fulfilled the inclusion criteria . Five groups of allergens were studied: group 1 (pollens), 2 (fungus), 3 (inhaled allergens), 4 (food) and 5 (other allergens), the value of seric IgE was registered too . RESULTS: The main allergens that were found are: 156 (43.8%) Amaranthus palmeri, 163 (45.5%) Candida albicans, 262 (73.6%) dust mite, 33 (9.3%) chicken and 4 (1.2%) apple/pineapple . The main diseases were: 254 (71.3%) allergic rhinitis, 52 (14.6%) asthma-rhinitis, 39 (11%) asthma and 63 (17.6%) others, the value of seric IgE was minimum of 1 and maximum of 9,620 Ul/dL . CONCLUSIONS: Allergens affecting patients are not different of what is reported in world-wide literature; however, it is of great significance to know this and take it as a guideline to achieve clinical improvement with the use of an appropriate immunotherapy.

Clin Exp Obstet Gynecol, 2004, 31(3), 175 - 8
Infections of the lower female genital tract during childhood and adolescence; Deligeoroglou E et al.; PURPOSE: To review the pathogenesis, clinical presentation, diagnosis and treatment of lower female genital tract infections at a young age . METHODS: Review study . CONCLUSIONS: Lower female genital tract infections at a young age may involve the vulva, the vagina and, less frequently, the fallopian tubes . Good knowledge of the physiology and anatomy of the respective areas plays an important role in the diagnosis and treatment of vulvovaginitis . Candida albicans is the most frequent cause of infection, while Gardnerella vaginalis, Chlamydia trachomatis, Mycoplasma, and Trichomonas vaginalis are rarer ones . The clinical presentation includes a variety of symptoms and signs, with vaginal discharge being the prominent one . Treatment should be causative after careful investigation while preventive advice is mandatory.

Clin Microbiol Rev, 2004 Oct, 17(4), 729 - 59, table of contents
Immunopathogenesis of oropharyngeal candidiasis in human immunodeficiency virus infection; de Repentigny L et al.; Oropharyngeal and esophageal candidiases remain significant causes of morbidity in human immunodeficiency virus (HIV)-infected patients, despite the dramatic ability of antiretroviral therapy to reconstitute immunity . Notable advances have been achieved in understanding, at the molecular level, the relationships between the progression of HIV infection, the acquisition, maintenance, and clonality of oral candidal populations, and the emergence of antifungal resistance . However, the critical immunological defects which are responsible for the onset and maintenance of mucosal candidiasis in patients with HIV infection have not been elucidated . The devastating impact of HIV infection on mucosal Langerhans' cell and CD4(+) cell populations is most probably central to the pathogenesis of mucosal candidiasis in HIV-infected patients . However, these defects may be partly compensated by preserved host defense mechanisms (calprotectin, keratinocytes, CD8(+) T cells, and phagocytes) which, individually or together, may limit Candida albicans proliferation to the superficial mucosa . The availability of CD4C/HIV transgenic mice expressing HIV-1 in immune cells has provided the opportunity to devise a novel model of mucosal candidiasis that closely mimics the clinical and pathological features of candidal infection in human HIV infection . These transgenic mice allow, for the first time, a precise cause-and-effect analysis of the immunopathogenesis of mucosal candidiasis in HIV infection under controlled conditions in a small laboratory animal.

Mycopathologia, 2004 Jul, 158(1), 39 - 41
Genotypic differences of Candida albicans and C . dubliniensis isolates related to ethnic/racial differences within the same geographic area; McCullough MJ et al.; Candida albicans and C . dubliniensis genotype differences among Israeli ethnic groups were assessed . Isolates from Jews (51), Arabs (35) and Druze (25) were genotyped . The distributions among ethnic groups were not different, however they differed (p = 0.002) from global populations . Therefore, C . albicans and C . dubliniensis genotype distribution differences in Israel are related to changes in all ethnic groups.

Mycopathologia, 2004 Jul, 158(1), 9 - 15
Enhancement of amphotericin B activity against Candida albicans by superoxide radical; Okamoto Y et al.; This study aimed to examine the involvement of oxidative damage in amphotericin B (AmB) activity against Candida albicans using the superoxide (O2-) generator paraquat (PQ) . The effects of PQ on AmB activities against growth, viability, membrane permeability and respiration were examined in a wild-type parent strain (K) and a respiration-deficient mutant (KRD-19) since PQ-induced superoxide generation depends on respiration . In the parent strain, the minimal inhibitory concentration (MIC) of AmB, 0.25 microg/ml, tested with a liquid culture was lowered to 0.025 microg/ml by 1 mM PQ . Such a PQ-induced decrease in the MIC value of AmB was minimal in the mutant . Similar PQ-induced enhancement of AmB activity toward the parent strain was also observed with growth on an agar medium . In viability tests, when candidal cells were exposed to AmB (0.1 microg/ml) for I h, the lethality of AmB was enhanced by 1 mM PQ only in the parent strain . Exogenous superoxide dismutase and catalase failed to diminish the enhancing effect of PQ on the growth inhibitory activity of AmB in the parent strain, suggesting an interaction between superoxide and AmB in candidal cells . The enhancement of AmB activity by PQ, observed preferentially in the wild-type strain, can be explained by extensive superoxide generation depending on respiration . These results suggest that oxidative damage induced by superoxide is involved in AmB activity against C . albicans.

Ital Heart J, 2004 Jul, 5(7), 541 - 7
Ten-year experience with cryopreserved aortic allografts in the surgical treatment of aortic valve pathologies; Rocco F et al.; BACKGROUND: The aim of this study was to evaluate the performance of cryopreserved aortic allografts (CAA) in the treatment of adult aortic valve pathologies . METHODS: Between May 1992 and October 2002, 122 CAA were implanted in 119 adult patients with pathologies of the aortic valve . The mean age of the patients was 38.03 +/- 13.6 years (range 17-78 years) . Thirty had had previous cardiac surgery . The principal indication was endocarditis (n = 45) . In 66 patients one or more associated pathologies were present including: an abscess of the left ventricular outflow tract (n = 32), an aneurysm of the ascending aorta (n = 22), mitral incompetence (n = 10), and coronary artery disease (n = 3) . The indications for surgery were elective in 77 cases and urgent in 45 . The CAA was implanted as a total root replacement in 46 patients and as a free-hand in 76 . In 66 patients an associated procedure such as a left ventricular outflow tract reconstruction (in 27 cases) was performed . RESULTS: The in-hospital mortality was 5.73% (7/122) . In one patient the CAA was replaced before discharge with another CAA because of a mediastinitis with endocarditis by Candida albicans . At the follow-up of the 114 patients discharged from the hospital (mean 50.11 months, range 1-126 months), 6 patients died and 6 were reoperated . The actuarial 10-year survival, reoperation-free, endocarditis-free, structural degeneration-free rates were respectively 83.88, 81.70, 86.30, and 92.80% . CONCLUSIONS: From our experience we conclude that CAA are good substitutes for aortic valve replacement and even in desperate situations exhibit an acceptable long-term performance.

J Antimicrob Chemother, 2004 Dec, 54(6), 999 - 1006 Epub 2004 Oct 14.
Regulated overexpression of CDR1 in Candida albicans confers multidrug resistance; Niimi M et al.; OBJECTIVES: Information on the function of Candida albicans ATP-binding cassette (ABC) membrane transporter Cdr1p has come from studying the effect of gene inactivation in C . albicans and from heterologous Cdr1p expression in the yeast Saccharomyces cerevisiae . These approaches, however, give only an indirect indication of Cdr1p function in C . albicans itself . The objective of this study was to determine Cdr1p function in C . albicans by induced overexpression of Cdr1p in a C . albicans CDR1-deleted strain . METHODS: The C . albicans CDR1 open reading frame was fused to the C . albicans HEX1 promoter and used to complement a CDR1-null mutant to create strain FL3 . The effect of inducing the FL3 HEX1 promoter, by growth on medium containing N-acetylglucosamine (GlcNAc) as the carbon source, on CDR1 expression and drug susceptibility was determined . RESULTS: C . albicans FL3 cells grown on medium containing GlcNAc overexpressed CDR1 mRNA and a 170 kDa plasma membrane protein that reacted with anti-Cdr1p antibodies . Overexpression of Cdr1p in C . albicans FL3 conferred resistance to structurally unrelated chemicals such as terbinafine, brefeldin A, cerulenin and nigericin as well as to azole antifungal agents, but not resistance to polyene antibiotics . FK506, ascomycin and ciclosporin A chemosensitized FL3 to fluconazole . FL3 cells grown on GlcNAc effluxed 5.3 times as much Cdr1p substrate rhodamine 6G, over a 10 min period, as FL3 cells grown on glucose, and this rhodamine 6G efflux was inhibited by including fluconazole in the assay . CONCLUSION: This study provides the first direct demonstration of Cdr1p pump activity in C . albicans.

J Biol Chem, 2004 Dec 31, 279(53), 55060 - 72 Epub 2004 Oct 13.
The TRK1 potassium transporter is the critical effector for killing of Candida albicans by the cationic protein, Histatin 5; Baev D et al.; The principal feature of killing of Candida albicans and other pathogenic fungi by the catonic protein Histatin 5 (Hst 5) is loss of cytoplasmic small molecules and ions, including ATP and K(+), which can be blocked by the anion channel inhibitor 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid . We constructed C . albicans strains expressing one, two, or three copies of the TRK1 gene in order to investigate possible roles of Trk1p (the organism's principal K(+) transporter) in the actions of Hst 5 . All measured parameters (Hst 5 killing, Hst 5-stimulated ATP efflux, normal Trk1p-mediated K(+) ((86)Rb(+)) influx, and Trk1p-mediated chloride conductance) were similarly reduced (5-7-fold) by removal of a single copy of the TRK1 gene from this diploid organism and were fully restored by complementation of the missing allele . A TRK1 overexpression strain of C . albicans, constructed by integrating an additional TRK1 gene into wild-type cells, demonstrated cytoplasmic sequestration of Trk1 protein, along with somewhat diminished toxicity of Hst 5 . These results could be produced either by depletion of intracellular free Hst 5 due to sequestered binding, or to cooperativity in Hst 5-protein interactions at the plasma membrane . Furthermore, Trk1p-mediated chloride conductance was blocked by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid in all of the tested strains, strongly suggesting that the TRK1 protein provides the essential pathway for ATP loss and is the critical effector for Hst 5 toxicity in C . albicans.

Yeast, 2004 Oct 15, 21(13), 1121 - 31
An in vitro assay for (1 --> 6)-beta-D-glucan synthesis in Saccharomyces cerevisiae; Vink E et al.; (1 --> 6)-beta-D-glucan is a key cell wall component of Saccharomyces cerevisiae and Candida albicans . Many genes are known to affect the levels or structure of this glucan, but their roles and a molecular description of the synthesis of (1 --> 6)-beta-D-glucan remain to be established and a method to measure (1 --> 6)-beta-D-glucan synthase activity in vitro would provide an enabling tool . Here, conditions for the detection of in vitro synthesis of this polymer are described . Crude membrane preparations from S . cerevisiae were isolated, and incubated in the presence of UDP-glucose and GTP . With anti-(1 --> 6)-beta-D-glucan-specific antibodies, a time-dependent increase in the amount of this glucan was demonstrated in a dot-blot assay, or through an inhibition enzyme immunoassay . Antibody specificity was validated by competition experiments using pustulan, a (1 --> 6)-beta-D-glucan, laminarin, a (1 --> 3)-beta-D-glucan, yeast mannan and glycogen . The identity of the reaction product was also demonstrated by its sensitivity to a recombinant (1 --> 6)-beta-D-glucanase . Extracts from mutants in 10 genes with a wide range of altered cell wall (1 --> 6)-beta-D-glucan levels were assayed for in vitro synthesis of the polymer . A strong correlation of in vitro synthase activity with in vivo glucan levels was found, providing genetic support for the specificity of the assay . The basis for the GTP-dependence of the synthase reaction was studied . Extracts from rho2, rho3, rho4 and rho5 null mutants had wild-type in vitro activity . In contrast, Rho1p overproduction led to increased in vitro synthesis, implicating Rho1p in the regulation of (1 --> 6)-beta-D-glucan synthesis . Copyright (c) 2004 John Wiley & Sons, Ltd.

Rev Iberoam Micol, 1997 Mar, 14(1), 26 - 8
{Contribution to the study of dermatomycosis in Catalonia.}; Boncompte E et al.; We report the results of a study which aim was the mycological identification of specimens coming from patients included in a clinical trial . A total of 445 specimens from patients with clinical diagnosis of dermatomicosis were processed during 8 months (138 pityriasis versicolor, 28 cutaneous candidosis and 279 dermatophytosis) . A 48% of pityriasis versicolor cultures were positive for Malassezia furfur, 50% of candidosis cultures were positive for yeasts and 67% of dermatophytosis cultures were positive for dermatophytes . According to our results Candida albicans was the principal causative agent for cutaneous candidosis and Trichophyton mentagrophytes and Trichophyton rubrum were the most frequent isolated species causing dermatophytosis.

Rev Iberoam Micol, 1997 Mar, 14(1), 23 - 5
Diagnosis of candidosis by amplification of small subunit of 18S rRNA gene; Unzaga MJ et al.; A PCR assay for the diagnosis of infection produced by Candida sp . was developed . The primers, designated 520 and 522, were selected from highly-conserved areas of the small subunit (ssu) 18S rRNA gene of Candida spp . To check the value of the results a Candida albicans oligonucleotide probe, digoxigenin-labeled, and a general Candida probe were used in hybridization experiments with the amplified products . We were able to detect a Candida- specific fragment of 1800bp from different clinical samples . The procedure described could provide an interesting complement to present diagnostic methods of detecting Candida sp in clinical samples.

Rev Iberoam Micol, 2001 Mar, 18(1), 17 - 22
Cell surface hydrophobicity-associated adherence of Candida dubliniensis to human buccal epithelial cells; Jabra-Rizk MA et al.; Microbial adherence to mucosal surfaces is an important first step in the initiation of the pathogenic process in the oral cavity . Candida albicans, the most adherent and pathogenic Candida species, utilizes a variety of mechanisms to adhere to human tissues . Although the strongest mechanism of adherence involves mannoprotein adhesins on C . albicans, cell surface hydrophobicity (CSH) plays an important role in the adherence process by providing hydrophobic interactions that turn the initial attachment between the yeast and a surface into a strong bond . Recent cell wall analytical and comparative studies showed that, Candida dubliniensis, unlike C . albicans, possesses cell surface variations that allow it to be constantly hydrophobic, regardless of growth temperature . Based on these observations, the present study was designed to compare the adherence abilities of C . dubliniensis and C . albicans to pooled human buccal epithelial cells (BEC), in regards to their cell surface hydrophobicity . Ten C . albicans and nine C . dubliniensis isolates, as well as the C . albicans hydrophobic variant A9V10 were evaluated for adherence with BEC using visual aggregation in the wells of a microtiter plate and microscopic examination . All 11 C . albicans isolates failed to show adherence to BEC, visually or microscopically, when grown at 37 degrees C . The same isolates, however, showed significant increase in aggregation and microscopic adherence to BEC when grown at 25 degrees C . All C . dubliniensis isolates tested and the A9V10 C . albicans hydrophobic variant resulted in visual aggregation and adhered to BEC when grown at either temperature . The findings from this study show that, based on comparative adherence results and growth temperature changes, C . dubliniensis seems to have greater adherence to BEC than do typical C . albicans strains and that hydrophobic interactions seem to be the mechanism of adherence involved . Although many questions remain to be answered regarding the clinical implications of this observed in vitro enhanced adherence of C . dubliniensis to human BEC, these findings support the establishment of this novel species as a clinically significant yeast.

Rev Iberoam Micol, 2001 Mar, 18(1), 12 - 6
UV and X-ray sensitivity of Candida albicans laboratory strains and mutants having chromosomal alterations; Janbon G et al.; Utilization of L-sorbose, D-arabinose or primary fluconazole resistance in Candida albicans are controlled by copy number of specific chromosomes . On the other hand, spontaneous morphological mutants have a wide range of chromosomal alterations . We have investigated the UV and X-ray sensitivity of these mutants, as well as C . albicans laboratory strains . While L-sorbose utilizing mutants had normal sensitivities, a large subclass of D-arabinose utilizing mutants was abnormally sensitive to UV . Spontaneous morphological mutants responded differently, an expected result because of the heterogeneous nature of their electrophoretic karyotypes . We suggest that the differences in UV and X-ray sensitivity are due to gene imbalance caused by some chromosomal alterations . In this respect, the radiation sensitivity is similar to other features impaired by changes in chromosomes, but is unlike the acquisition of the ability to utilize alternative nutrients or the acquisition of resistance to fluconazole . Our studies also revealed that strains of C . albicans heterozygous for the mating type loci exhibited the same X-ray sensitivity as homozygous or hemizygous strains, a finding which is in contrast to the properties of Saccharomyces cerevisiae, where heterozygous strains are more resistant . This feature of C . albicans strains may be indicative of an inefficient repair system that may be related to inefficiency of mating.

Rev Iberoam Micol, 2001 Mar, 18(1), 6 - 11
Differences in the Candida albicans antigenic expression after heat shock and infection; Calcedo R et al.; Heat-shock and infection induce changes in protein expression in C . albicans . To investigate if these alterations induce changes in antigenicity, we have compared the reactivity mediated by IgA antibodies of protein extracts from a strain of C . albicans and the same strain recovered from an infected animal, both at 24 degrees C and 37 degrees C . The antigenic variability was detected mainly in antigens recognized by salivary IgA . Antigens of 223, 205, 180 and 140 kDa were over-expressed in both strains at 37 degrees C, indicating that variations due to heat shock were present before and after infection . The antigens were characterized as mannoproteins located at the outer side of the cell wall . An antigen of 61 kDa was also detected in which the expression decreased significantly after infection This was independent of heat shock.

Phytother Res, 2004 Sep, 18(9), 754 - 7
Antibacterial activity of volatile component and various extracts of Spirulina platensis; Ozdemir G et al.; The methanol, dichloromethane, petroleum ether, ethyl acetate extracts and volatile components of Spirulina platensis were tested in vitro for their antimicrobial activity (four Gram-positive, six Gram-negative bacteria and Candida albicans ATCC 10239) . GC-MS analysis of the volatile components of S . platensis resulted in the identification of 15 compounds which constituted 96.45% of the total compounds . The volatile components of S . platensis consisted of heptadecane (39.70%) and tetradecane (34.61%) as major components . The methanol extract showed more potent antimicrobial activity than dichloromethane, petroleum ether, ethyl acetate extracts and volatile components . Copyright (c) 2004 John Wiley & Sons, Ltd.

Phytother Res, 2004 Sep, 18(9), 783 - 4
Antimicrobial activity of tiger's betel (Piper porphyrophyllum N.E . Br., Piperaceae); Wiart C et al.; The ethanol extract of leaves of Piper porphyrophyllum N.E . Br . showed a broad spectrum of antibacterial activity . The activity was increased on fractionation (hexane, dichloromethane and aqueous), particularly in the aqueous fraction . No activity was shown against tested Candida albicans . Copyright (c) 2004 John Wiley & Sons, Ltd.

Stomatologiia (Mosk), 2004, 83(5), 14 - 6
{Candida albicans adhesion to plastic during correction of removable dentures}; Antifungal treatment with carvacrol and eugenol of oral candidiasis in immunosuppressed rats; Fes Laboratoire de Biotechnologie, Faculte des Sciences, BP 1796 Atlas FES, MoroccoCarvacrol and eugenol, the main (phenolic) components of essential oils of some aromatic plants, were evaluated for their therapeutic efficacy in the treatment of experimental oral candidiasis induced by Candida albicans in immunosuppressed rats . This anticandidal activity was analyzed by microbiological and histopathological techniques, and it was compared with that of nystatin, which was used as a positive control . Microbiologically, carvacrol and eugenol significantly (p<0.05) reduced the number of colony forming units (CFU) sampled from the oral cavity of rats treated for eight consecutive days, compared to untreated control rats . Treatment with nystatin gave similar results . Histologically, the untreated control animals showed numerous hyphae on the epithelium of the dorsal surface of the tongue . In contrast no hyphal colonization of the epithelium was seen in carvacrol-treated animals, while in rats treated with eugenol, only a few focalized zones of the dorsal surface of the tongue were occupied by hyphae . In the nystatin treated group, hyphae were found in the folds of the tongue mucosa . Thus, the histological data were confirmed by the microbiological tests for carvacrol and eugenol, but not for the nystatin-treated group . Therefore, carvacrol and eugenol could be considered as strong antifungal agents and could be proposed as therapeutic agents for oral candidiasis.

J Infect, 2004 Nov, 49(4), 317 - 23
Predictors of adverse outcome from candidal infection in a tertiary care hospital; Ben-Abraham R et al.; OBJECTIVES: To retrospectively delineate predictors of adverse outcome by looking at the demographic features, therapy and outcome of systemic candida infection in a large tertiary care university-affiliated medical center . METHODS: We reviewed the clinical data on 186 inpatients with candidemia over a 6-year period . The major reason for their hospital admission was an underlying malignancy or an infection other than candidemia . RESULTS: Candida albicans, tropicalis, parapsilosis, glabrata and krusei caused 54, 22, 13, 8 and 3% of the candidemia episodes, respectively . The overall mortality was 42% and it was highest in patients suffering from candidemia of the glabrata species (73%) . Forty-eight (63%) of the 76 patients who received no anti-fungal treatment died compared to 38 (34%) of 110 patients who were treated (P < 0.05) . Predictors of adverse outcome were intensive care unit stay, renal failure, thrombocytopenia and the need for mechanical ventilation or inotropic support . CONCLUSIONS: We identified four predictors of mortality from candidemia infection . Their validity should be further assessed and the specific candida strains and their susceptibility need to be methodically identified . Our data support immediate initiation of therapy at first identification of infection.

Gene, 2004 Oct 27, 341, 119 - 27
The SAT1 flipper, an optimized tool for gene disruption in Candida albicans; Reuss O et al.; The construction of Candida albicans mutants by targeted gene disruption usually depends on the use of nutritional markers for the selection of prototrophic transformants from auxotrophic host strains, but it is becoming increasingly evident that this strategy may cause difficulties in the interpretation of mutant phenotypes . Here, we describe a new method for inactivating both alleles of a target gene in C . albicans wild-type strains to obtain homozygous null mutants . The SAT1 flipping method relies on the use of a cassette that contains a dominant nourseothricin resistance marker (caSAT1) for the selection of integrative transformants and a C . albicans-adapted FLP gene that allows the subsequent excision of the cassette, which is flanked by FLP target sequences, from the genome . Two rounds of integration/excision generate homozygous mutants that differ from the wild-type parent strain only by the absence of the target gene, and reintegration of an intact gene copy for complementation of mutant phenotypes is performed in the same way . Transformants are obtained after only 1 day of growth on a selective medium, and integration into the target locus occurs with high specificity after adding homologous flanking sequences on both sides of the cassette . FLP-mediated excision of the SAT1 flipper cassette is achieved by simply growing the transformants for a few hours in medium without selective pressure, and nourseothricin-sensitive (NouS) derivatives can easily be identified by their slower growth on indicator plates containing a low concentration of nourseothricin . We demonstrate the use of the system by deleting the OPT1 gene, which encodes an oligopeptide transporter, in the C . albicans model strain SC5314 . The null mutants became resistant to the toxic peptide KLLEth, and reintroduction of an intact OPT1 copy restored susceptibility . The SAT1 flipping method provides a highly efficient method for gene disruption in C . albicans wild-type strains, which eliminates currently encountered problems in the genetic analysis of this important human fungal pathogen.

Arch Pharm Res, 2004 Sep, 27(9), 885 - 92
Synthesis and antimicrobial activity of novel tetrahydrobenzothienopyrimidines; Eissa AA et al.; Due to the rapidly growing number of resistant strains of bacteria, the search for antibacterial agents with new modes of action will always remain an important and challenging task . Thus, the reaction of 2-substituted or unsubstituted-4-(4-acetylanilino)-5,6,7,8-tetrahydrobenzo{b}thieno{2,3-d}pyrimidine derivatives 1-3 with the hydrazine derivatives, semi and/or thiosemicarbazides, provided the corresponding hydrazones 4-6 and semi and/or thiosemicarbazones 7-9 . Claisen-Schmidt condensation of compounds 1 or 2 with the appropriate aldehyde yielded the chalcones 10, 11 which, when treated with hydroxylamine hydrochloride gave rise to the isoxazoline-containing compounds 12, 13 . Furthermore, reacting the respective chalcones 10, 11 with different hydrazines, urea and/or thiourea, furnished compounds 14, 15, 16, and 17 respectively . Representative compounds were tested for their antimicrobial activity against Candida albicans and certain gram-positive and gram-negative bacteria . Their MICs were then determined . Compound 15e, showed a broad spectrum of activity while most of the other compounds showed varying antimicrobial activity.

Med Mycol, 2004 Aug, 42(4), 385 - 9
Ergosterol gene expression in wild-type and ergosterol-deficient mutants of Candida albicans; Pierson CA et al.; The ergosterol pathway is the major target of the azole antifungals . We have developed a panel of five viable ergosterol biosynthetic mutants (erg2, erg3, erg6, erg11 and erg24) and have performed Northern analyses to study transcriptional regulation using probes to four ergosterol biosynthetic genes (ERG2, ERG7, ERG11 and ERG25), as well as probes to two additional genes encoding ergosterol cytochrome coenzymes (CYB5 and NCP1) . ERG11, which encodes the sterol 14-demethylase, the direct target of the azole antifungals, was the most up-regulated gene followed by ERG7 and ERG25 . Transcription of the four ergosterol genes was most up-regulated in erg24 and erg6 mutant backgrounds, deficient in C-14 reductase and the C-24 sterol transmethylase, respectively . Unexpectedly, we also found that the two cytochrome genes, CYB5 encoding cytochrome b5 and NCP1 encoding the cytochrome P450 reductase, were not regulated markedly different from wild-type in the erg2, erg3, erg6, erg11 and erg24 strains of Candida albicans.

Med Mycol, 2004 Aug, 42(4), 373 - 8
Wound infection due to Absidia corymbifera and Candida albicans with fatal outcome; Horre R et al.; A case of a mixed infection due to Candida albicans and the zygomycete Absidia corymbifera in a 38-year-old, previously healthy, Caucasian male is presented . The infection developed following serial rib fractures, and ruptures of kidney, liver and biliary tract as well as a pancreatic contusion resulting from a traffic accident . During intensive care treatment the patient underwent several surgical procedures but subsequently experienced multi-organ failure and sepsis . Some weeks later, fungal growth was observed macroscopically on the patient's skin and wounds . From wound swabs C . albicans and A . corymbifera were grown . Histopathology of abdominal tissue yielded pseudohyphae and coenocytic hyphae . Although surgical debridement and antifungal treatment with amphotericin B and 5-flucytosine were started immediately, the patient died in therapy-refractory septic multi-organ failure.

Med Mycol, 2004 Aug, 42(4), 341 - 8
Apolipoprotein-E-deficient mice exhibit an increased susceptibility to disseminated candidiasis; Alieke GV et al.; The effect of hyperlipoproteinemia on systemic candidiasis was investigated by assessing the susceptibility of hyperlipoproteinemic, apolipoprotein E (ApoE)-deficient (ApoE -/-) mice to a systemic Candida albicans infection . The absence of ApoE in these mice resulted in an eightfold increase in plasma lipoprotein concentrations in the very low-density lipoprotein (VLDL) fraction, as compared with levels seen in ApoE +/+ mice . Mortality due to candidemia was significantly higher (86%) in ApoE -/- mice than in ApoE+/+ mice (52%), and in platings of homogenized kidney material on fungal culture medium, ApoE -/- mice yielded significantly higher levels of C . albicans outgrowth than did ApoE+/+ mice . C albicans grew twofold better in ApoE -/- plasma in 4 h than in ApoE+/+ plasma, and depletion of lipoproteins from plasma resulted in a significant seven- to tenfold increase in C . albicans growth . Recombinant ApoE did not directly inhibit C . albicans growth . Our data indicate that the increased susceptibility of ApoE -/- mice to C albicans is due both to increased growth of blastoconidia in ApoE -/- mice in response to the availability of lipids as nutrients, and to the neutralization of candidacidal factors by lipoproteins . This study suggests that lipoproteins play a significant role in host defense against candidiasis.

Med Mycol, 2004 Aug, 42(4), 319 - 24
Immunogenicity and protective effect of recombinant enolase of Candida albicans in a murine model of systemic candidiasis; Montagnoli C et al.; Enolase, a 46-kDa glycolytic enzyme, is an immunodominant antigen of the opportunistic pathogen Candida albicans . A recombinant 6 x His-tagged enolase was studied, in conjunction with interleukin-12 (IL-12), as an adjuvant for cytokine induction favouring protection in a murine model of haematogenous candidiasis . Mice immunized with enolase plus IL-12 showed increased antibody titres against enolase, as well as increased median survival time and decreased fungal burden in kidneys, in comparison to non-immunized or IL-12-treated mice . This increased survival was attributable to enolase-induced cell-mediated immunity as it also occurred in B-cell-deficient mice . Enolase immunization stimulated a predominant T-helper-1 (Th1) cytokine pattern in splenic cells and induced production of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) by purified CD4+ T cells . However, despite the elevation of immunogenicity, recombinant enolase induced only a modest protection against disseminated candidiasis, suggesting a form of protection likely attributable to the induction of a Th1 cell-mediated immune response.

Med Mycol, 2004 Aug, 42(4), 305 - 9
Safety aspects of working with Candida albicans-infected mice; MacCallum DM et al.; When the opportunity arose in the course of four experiments with mice and one with guinea pigs, all systemically infected with Candida albicans, the animals' bedding, work surfaces, surrounding walls, the balance pan and tools used in homogenization of tissues were sampled with contact plates or by water washing for the presence of viable C . albicans cells . Although substantial viable counts of C . albicans were measured in homogenized samples of kidneys and other tissues, no colonies of the fungus were recovered at any time from the work surfaces, walls or homogenizer stand . Contact samples of the homogenizer dispersal tool made on four occasions during the course of 24 successive homogenizations showed that few viable C . albicans could be cultured from the tool after two water washes, and none at all after two washes with 70% ethanol . Water samples of the contents of three cages that had housed infected mice were all negative for viable C . albicans, however, direct contact plate samples of the bedding material and excreta in seven cages yielded positive cultures with colony counts from 1 to 8 per sample in five instances and 18 in one instance . It is concluded that the potential infection risk to personnel of working with this hazard group 2 fungus is minimal and the highly stringent safety regulations for all organisms in hazard group 2 may err on the side of over-caution.

Med Mycol, 2004 Aug, 42(4), 293 - 304
Assessment of Candida albicans genes expressed during infections as a tool to understand pathogenesis; Nguyen MH et al.; Candida albicans is the most common fungal opportunistic pathogen of humans and causes mucocutaneous, bloodstream and deep organ infections . Screening for C . albicans genes that are preferentially expressed within infected hosts represents a strategy to identify novel virulence factors and define global expression patterns relevant to pathogenesis . Until recently, C . albicans has not been amenable to screening using existing technologies . This has begun to change with the development of new molecular genetic tools and the sequencing of the C . albicans genome . In this paper, we review studies using recently developed techniques to identify genes expressed by C . albicans during infections, as well as work from our laboratory using a human antibody-based strategy . Along with others, we have shown that selected in vivo expressed genes encode known and previously unrecognized candidal virulence factors . Future studies in this area will identify additional novel virulence factors, as well as advance our understanding of pathogenesis.

J Clin Microbiol, 2004 Oct, 42(10), 4796 - 8
Tobacco agar, a new medium for differentiating Candida dubliniensis from Candida albicans; Khan ZU et al.; Isolates of Candida dubliniensis may be misidentified as Candida albicans in microbiological laboratories if only the germ tube and/or the chlamydospore test is used for identification to the species level . In this study, we have evaluated the efficacy of tobacco agar for the differentiation of C . dubliniensis from C . albicans . On this medium at 28 degrees C, all 30 C . dubliniensis isolates produced yellowish-brown colonies with hyphal fringes and abundant chlamydospores, whereas 54 C . albicans isolates formed smooth, white-to-cream-colored colonies with no chlamydospore production . This medium provides a simple tool for presumptive differentiation of C . dubliniensis from C . albicans.

J Clin Microbiol, 2004 Oct, 42(10), 4726 - 34
Differential expression of secretory aspartyl proteinase genes (SAP1-10) in oral Candida albicans isolates with distinct karyotypes; Tavanti A et al.; Two karyotypes of oral Candida albicans isolates, named b and c, constituted >80% of a collection from healthy carriers (22 b and 16 c isolates) and oral candidiasis patients who were either infected (31 b and 16 c isolates) or uninfected (13 b and 38 c isolates) with human immunodeficiency virus (HIV) . The prevalence of the b and c karyotypes within HIV-positive and HIV-negative patients, respectively, who were suffering from oral candidiasis (P < or = 0.0001) suggested that these two types possessed different virulence potentials . Since C . albicans proteinases (Saps) are virulence factors in oral candidiasis, we evaluated whether the b and c karyotypes secreted different levels of Saps and expressed different patterns of Sap-encoding genes (SAP1-10) . We found that the mean value of Sap activity was significantly lower (P = 0.003) in the commensal type than in the infectious b karyotype, whereas Sap activity in the commensal c type was as high as that registered for the infectious c strains . Marked differences in SAP mRNA expression were observed in commensal strains under non-Sap-inducing conditions, with all SAP genes being expressed only by strains with the c karyotype; interestingly, none of the commensal b strains expressed SAP2 . In addition, while all of the SAP1-10 genes were detectable under Sap-inducing conditions, the timing of their expression during growth differed significantly, with mRNAs of SAP1-10 genes detected at 8 and 24 h postinoculation in c and b commensal strains, respectively . This provides the first evidence that commensal oral C . albicans isolates with distinct karyotypes are characterized by different patterns of SAP1-10 gene expression and different levels of Sap secretion.

Eukaryot Cell, 2004 Oct, 3(5), 1272 - 86
Functional characterization of myosin I tail regions in Candida albicans; Oberholzer U et al.; The molecular motor myosin I is required for hyphal growth in the pathogenic yeast Candida albicans . Specific myosin I functions were investigated by a deletion analysis of five neck and tail regions . Hyphal formation requires both the TH1 region and the IQ motifs . The TH2 region is important for optimal hyphal growth . All of the regions, except for the SH3 and acidic (A) regions that were examined individually, were required for the localization of myosin I at the hyphal tip . Similarly, all of the domains were required for the association of myosin I with pelletable actin-bound complexes . Moreover, the hyphal tip localization of cortical actin patches, identified by both rhodamine-phalloidin staining and Arp3-green fluorescent protein signals, was dependent on myosin I . Double deletion of the A and SH3 domains depolarized the distribution of the cortical actin patches without affecting the ability of the mutant to form hyphae, suggesting that myosin I has distinct functions in these processes . Among the six myosin I tail domain mutants, the ability to form hyphae was strictly correlated with endocytosis . We propose that the uptake of cell wall remodeling enzymes and excess plasma membrane is critical for hyphal formation.

Eukaryot Cell, 2004 Oct, 3(5), 1164 - 8
Pmt-mediated O mannosylation stabilizes an essential component of the secretory apparatus, Sec20p, in Candida albicans; Weber Y et al.; Sec20p is an essential endoplasmic reticulum (ER) membrane protein in yeasts, functioning as a tSNARE component in retrograde vesicle traffic . We show that Sec20p in the human fungal pathogen Candida albicans is extensively O mannosylated by protein mannosyltransferases (Pmt proteins) . Surprisingly, Sec20p occurs at wild-type levels in a pmt6 mutant but at very low levels in pmt1 and pmt4 mutants and also after replacement of specific Ser/Thr residues in the lumenal domain of Sec20p . Pulse-chase experiments revealed rapid degradation of unmodified Sec20p (38.6 kDa) following its biosynthesis, while the stable O-glycosylated form (50 kDa) was not formed in a pmt1 mutant . These results suggest a novel function of O mannosylation in eukaryotes, in that modification by specific Pmt proteins will prevent degradation of ER-resident membrane proteins via ER-associated degradation or a proteasome-independent pathway.

Eukaryot Cell, 2004 Oct, 3(5), 1154 - 63
Inner kinetochore of the pathogenic yeast Candida glabrata; Stoyan T et al.; The human pathogenic yeast Candida glabrata is the second most common Candida pathogen after Candida albicans, causing both bloodstream and mucosal infections . The centromere (CEN) DNA of C . glabrata (CgCEN), although structurally very similar to that of Saccharomyces cerevisiae, is not functional in S . cerevisiae . To further examine the structure of the C . glabrata inner kinetochore, we isolated several C . glabrata homologs of S . cerevisiae inner kinetochore protein genes, namely, genes for components of the CBF3 complex (Ndc10p, Cep3p, and Ctf13p) and genes for the proteins Mif2p and Cse4p . The amino acid sequence identities of these proteins were 32 to 49% relative to S . cerevisiae . CgNDC10, CgCEP3, and CgCTF13 are required for growth in C . glabrata and are specifically found at CgCEN, as demonstrated by chromatin immunoprecipitation experiments . Cross-complementation experiments revealed that the isolated genes, with the exception of CgCSE4, are species specific and cannot functionally substitute for the corresponding genes in S . cerevisiae deletion strains . Likewise, the S . cerevisiae CBF3 genes NDC10, CEP3, and CTF13 cannot functionally replace their homologs in C . glabrata CBF3 deletion strains . Two-hybrid analysis revealed several interactions between these proteins, all of which were previously reported for the inner kinetochore proteins of S . cerevisiae . Our findings indicate that although many of the inner kinetochore components have evolved considerably between the two closely related species, the organization of the C . glabrata inner kinetochore is similar to that in S . cerevisiae.

Eukaryot Cell, 2004 Oct, 3(5), 1111 - 23
Msn2- and Msn4-like transcription factors play no obvious roles in the stress responses of the fungal pathogen Candida albicans; Nicholls S et al.; In Saccharomyces cerevisiae, the (C2H2)2 zinc finger transcription factors Msn2 and Msn4 play central roles in responses to a range of stresses by activating gene transcription via the stress response element (STRE; CCCCT) . The pathogen Candida albicans displays stress responses that are thought to help it survive adverse environmental conditions encountered within its human host . However, these responses differ from those in S . cerevisiae, and hence we predicted that the roles of Msn2- and Msn4-like proteins might have been functionally reassigned in C . albicans . C . albicans has two such proteins: CaMsn4 and Mnl1 (for Msn2- and Msn4-like) . CaMSN4, but not MNL1, weakly complemented the inability of an S . cerevisiae msn2 msn4 mutant to activate a STRE-lacZ reporter . Also, the disruption of CaMsn4 and Mnl1 had no discernible effect upon the resistance of C . albicans to heat, osmotic, ethanol, nutrient, oxidative, or heavy-metal stress or upon the stress-activated transcriptome in C . albicans . Furthermore, although Cap1-dependent activation of a Yap response element-luciferase reporter was observed, a STRE reporter was not activated in response to stresses in C . albicans . Ectopic expression of CaMsn4 or Mnl1 did not affect the cellular or molecular responses of C . albicans to stress . Under the conditions tested, the putative activation and DNA binding domains of CaMsn4 did not appear to be functional . These data suggest that CaMsn4 and Mnl1 do not contribute significantly to stress responses in C . albicans . The data are consistent with the idea that stress signaling in this fungus has diverged significantly from that in budding yeast.

Eukaryot Cell, 2004 Oct, 3(5), 1076 - 87
Transcriptional response of Candida albicans upon internalization by macrophages; Lorenz MC et al.; The opportunistic fungal pathogen Candida albicans is both a benign gut commensal and a frequently fatal systemic pathogen . The interaction of C . albicans with the host's innate immune system is the primary factor in this balance; defects in innate immunity predispose the patient to disseminated candidiasis . Because of the central importance of phagocytic cells in defense against fungal infections, we have investigated the response of C . albicans to phagocytosis by mammalian macrophages using genomic transcript profiling . This analysis reveals a dramatic reprogramming of transcription in C . albicans that occurs in two successive steps . In the early phase cells shift to a starvation mode, including gluconeogenic growth, activation of fatty acid degradation, and downregulation of translation . In a later phase, as hyphal growth enables C . albicans to escape from the macrophage, cells quickly resume glycolytic growth . In addition, there is a substantial nonmetabolic response imbedded in the early phase, including machinery for DNA damage repair, oxidative stress responses, peptide uptake systems, and arginine biosynthesis . Further, a surprising percentage of the genes that respond specifically to macrophage contact have no known homologs, suggesting that the organism has undergone substantial evolutionary adaptations to the commensal or pathogen lifestyle . This transcriptional reprogramming is almost wholly absent in the related, but nonpathogenic, yeast Saccharomyces cerevisiae, suggesting that these large-scale and coordinated changes contribute significantly to the ability of this organism to survive and cause disease in vivo.

Microbiology, 2004 Oct, 150(Pt 10), 3363 - 82
Comparative genomics using Candida albicans DNA microarrays reveals absence and divergence of virulence-associated genes in Candida dubliniensis; Moran G et al.; Candida dubliniensis is a pathogenic yeast species closely related to Candida albicans . However, it is less frequently associated with human disease and displays reduced virulence in animal models of infection . Here comparative genomic hybridization was used in order to assess why C . dubliniensis is apparently less virulent than C . albicans . In these experiments the genomes of the two species were compared by co-hybridizing C . albicans microarrays with fluorescently labelled C . albicans and C . dubliniensis genomic DNA . C . dubliniensis genomic DNA was found to hybridize reproducibly to 95.6 % of C . albicans gene-specific sequences, indicating a significant degree of nucleotide sequence homology (> 60 %) in these sequences . The remaining 4.4 % of sequences (representing 247 genes) gave C . albicans/C . dubliniensis normalized fluorescent signal ratios that indicated significant sequence divergence (< 60 % homology) or absence in C . dubliniensis . Sequence divergence was identified in several genes (confirmed by Southern blot analysis and sequence analysis of PCR products) with putative virulence functions, including the gene encoding the hypha-specific human transglutaminase substrate Hwp1p . Poor hybridization of C . dubliniensis genomic DNA to the array sequences for the secreted aspartyl proteinase-encoding gene SAP5 also led to the finding that SAP5 was absent in C . dubliniensis and that this species possesses only one gene homologous to SAP4 and SAP6 of C . albicans . In addition, divergence and absence of sequences in several gene families was identified, including a family of HYR1-like GPI-anchored proteins, a family of genes homologous to a putative transcriptional activator (CTA2) and several ALS genes . This study has confirmed the close relatedness of C . albicans and C . dubliniensis and has identified a subset of unique C . albicans genes that may contribute to the increased prevalence and virulence of this species.

Microbiology, 2004 Oct, 150(Pt 10), 3341 - 54
The GPI-anchored protein CaEcm33p is required for cell wall integrity, morphogenesis and virulence in Candida albicans; Martinez-Lopez R et al.; Ecm33p is a widely distributed fungal protein with functional relevance, clearly demonstrated by ecm33Delta mutant phenotypes, mainly related to the cell wall . Homology searches with Saccharomyces cerevisiae genes identified Candida albicans Ecm33p, as well as the two other proteins of its family: Pst1p and the product of YCL048w . C . albicans Ecm33p is a 423 aa protein which has the typical features of cell-surface GPI proteins and is able to complement S . cerevisiae ecm33Delta cell wall defects . Heterozygous (RML1) and homozygous (RML2) mutants of CaECM33 were obtained, as well as a single and a double reintegrant (RML3 and RML4, respectively) . Caecm33 mutant strains displayed an aberrant morphology, being more rounded and bigger than the wild-type, suggesting morphogenetic defects . They also exhibited cell wall defects, with enhanced sensitivity to different compounds that interfere in polymerization of cell wall components (Calcofluor white, Congo red and hygromycin B) and a marked tendency to flocculate extensively . In addition, CaEcm33p is required for normal C . albicans yeast-to-hyphae transition in vitro . In liquid medium (5 % serum), the transition was delayed in Caecm33 mutants, and after 24 h the culture contained very abnormal large and rounded cells . On solid medium (10 % serum, Spider or SLADH) RML2 failed to produce hyphae and media invasiveness . CaECM33 showed a gene dosage effect, demonstrated by the intermediate phenotype of the heterozygous mutants RML1 and confirmed by Northern blot analysis . Furthermore, CaEcm33p is also involved in C . albicans virulence . In a murine systemic model of infection, 100 % mouse survival and no kidney or brain colonization were obtained 30 days after infection with 10(6) Candida cells of any homozygous or heterozygous Caecm33Delta mutant tested . In contrast, all mice infected with parental or RML4 (two CaECM33 copies reintegrated) strains died in a few days, showing that, in these conditions, two CaECM33 copies were required for virulence.

Microbiology, 2004 Oct, 150(Pt 10), 3305 - 13
Studies on the regulation of the two-component histidine kinase gene CHK1 in Candida albicans using the heterologous lacZ reporter gene; Li D et al.; The two-component histidine kinase Chk1p of Candida albicans has been implicated in the regulation of cell wall biosynthesis . Deletion of CHK1 results in avirulence that in part may be due to the increased sensitivity of mutant strains to polymorphonuclear leukocytes . The mutant also does not adhere to human oesophageal tissue in vitro, probably as a consequence of its altered cell wall . In the current study, a CHK1 promoter-lacZ reporter (CHK1prlacZ) construct was expressed in wild-type C . albicans strain CAI4 and in two-component signal transduction mutants to determine the effect of environmental stress conditions on the regulation of CHK1 and the co-regulatory activities among these proteins . It is shown that lacZ expression varied according to the type of growth conditions and incubation time; expression was also influenced by the strain background . lacZ expression in CAI4 was greater at 37 degrees C and at a pH of 3.5 and in the presence of 4 mM H2O2, 0.1 mM menadione, 10 % serum or 1.5 M NaCl compared to cells grown at 30 or 42 degrees C . The increases in expression were time-dependent and not observed until cells were incubated for 120 min in these conditions (P < 0.05) . As a correlate of the increase in transcription of CHK1-lacZ in the presence of H2O2, the chk1 mutant was more sensitive than wild-type and revertant cells to H2O2 in vitro . In addition to strain CAI4, we also measured CHK1p-lacZ reporter activity of mutants deleted in genes encoding other two-component proteins such as the response regulator gene SSK1, the histidine kinases, SLN1 and NIK1, and the HOG1 MAP kinase . Of these proteins, Ssk1p and Sln1p are presumed to mediate phosphotransfer to the HOG1 {hyperosmotic glycerol} MAP kinase pathway during oxidative and perhaps osmotic stress in C . albicans . Compared to strain CAI4, lacZ reporter activity increased significantly in the ssk1 mutant under all growth conditions after a 10 and 120 min incubation (P < 0.0001) . lacZ expression in the ssk1 mutant was less at 42 degrees C compared to all other growth conditions (P < 0.05) . Furthermore, lacZ reporter activity also increased in the hog1 mutant of C . albicans . These data suggest that SSK1 and HOG1 indirectly or directly negatively regulate CHK1 under most growth conditions tested . In the sln1 mutant, downregulation of CHK1 was observed in all growth conditions compared to strain CAI4 (P < 0.05), while regulation of lacZ in the nik1 mutant was similar to strain CAI4 except when cells were incubated in the presence of 4 mM H2O2 for 120 min (P < 0.05) . Western blot analysis was used to determine the role of Chk1p in phosphorylation of Hog1p under oxidative or osmotic stress . It was found that Hog1p was phosphorylated in the chk1 mutant similar to wild-type CAF2-1 cells, although the temporal events of phosphorylation differed slightly in mutant cells . These results show that transcription of CHK1, as measured by the lacZ reporter assay, is statistically increased when cells are exposed to several types of stress or when incubated in 10 % serum in a mutant-specific background and at a specific time point . Of importance, our data also suggest that lacZ expression is indirectly or directly regulated by the HOG1 MAP kinase pathway, although a determination of its position in this pathway or in a cross-talking pathway awaits additional studies.

Microbiology, 2004 Oct, 150(Pt 10), 3243 - 52
Candida albicans mutants in the BNI4 gene have reduced cell-wall chitin and alterations in morphogenesis; Rowbottom L et al.; The Candida albicans BNI4 gene was identified by homology to the Saccharomyces cerevisiae orthologue and encodes a predicted 1655 amino acid protein . In S . cerevisiae most cell-wall chitin is associated with primary septum formation and Bni4p is involved in tethering the Chs3p chitin synthase enzyme to the mother-bud neck by forming a bridge between a regulatory protein Chs4p and the septin Cdc10p . CaBni4p shows 20 % overall identity to the ScBni4p, with 73 % identity over the C-terminal 63 amino acids, which includes a putative protein phosphatase type 1 (PP1) binding domain . Northern blot analysis revealed a transcript of the expected size that was expressed in both yeast and hyphal growth forms . C . albicans has more chitin in its cell wall than S . cerevisiae, and again most chitin is synthesized by CaChs3p . The function of CaBNI4 was investigated by performing a targeted gene disruption using the 'Ura-blaster' method to delete amino acids 1120-1611 that are essential for function . The resulting Cabni4Delta/Cabni4Delta null mutants formed lemon-shaped yeast cells and had a 30 % reduction in cell-wall chitin, reduced hyphal formation on solid serum-containing medium and increased sensitivity to SDS and increased resistance to Calcofluor White . The Cabni4Delta/Cabni4Delta null mutants were unaffected in chitin ring formation, but often exhibited displaced bud sites with more obvious but flattened birth scars . Therefore, unlike in S . cerevisiae, the Cabni4 mutant apparently alters chitin distribution throughout the cell wall and not exclusively at the bud-neck region.

Microbiology, 2004 Oct, 150(Pt 10), 3151 - 61
Role of Pir1 in the construction of the Candida albicans cell wall; Martinez AI et al.; Searches in a Candida albicans database identified two Individual Protein Files (IPF 15363 and 19968) whose deduced amino acid sequences showed 42 % and 45 % homology with Saccharomyces cerevisiae Pir4 . The two DNA sequences are alleles of the same gene (CaPIR1) but IPF 19968 has a deletion of 117 bases . IPF 19968 encodes a putative polypeptide of 364 aa, which is highly O-glycosylated and has an N-mannosylated chain, four cysteine residues and seven repeats . Both alleles are expressed under different growth conditions and during wall construction by regenerating protoplasts . The heterozygous mutant cells are elongated, form clumps of several cells and are hypersensitive to drugs that affect cell wall assembly . CaPir1 was labelled with the V5 epitope and found linked to the 1,3-beta-glucan of the C . albicans wall and also by disulphide bridges when expressed in S . cerevisiae.

Microbiology, 2004 Oct, 150(Pt 10), 3115 - 28
Deficiencies in the essential Smp3 mannosyltransferase block glycosylphosphatidylinositol assembly and lead to defects in growth and cell wall biogenesis in Candida albicans; Grimme SJ et al.; Glycosylphosphatidylinositols (GPIs) are essential for viability in yeast and have key roles in cell wall construction . Assembly of Saccharomyces cerevisiae GPIs includes the addition of a fourth, side-branching mannose (Man) to the third Man of the core GPI glycan by the Smp3 mannosyltransferase . The SMP3 gene from the human pathogenic fungus Candida albicans has been cloned . CaSMP3 complements the inviable S . cerevisiae smp3 null mutant and, when expressed in an S . cerevisiae smp3/gpi13 double mutant, it permits in vivo conversion of the Man3-GPI precursor that accumulates in that mutant to a Man4-GPI . One allele of CaSMP3 was disrupted using the ura-blaster procedure, then the remaining allele was placed under the control of the glucose-repressible MAL2 promoter . Repression of CaSMP3 expression leads to accumulation of a GPI precursor glycolipid whose glycan headgroup contains three mannoses and bears a phosphodiester-linked substituent on its first Man . Under repressing conditions, cells exhibited morphological and cell wall defects and became inviable . CaSmp3p therefore adds a fourth, alpha1,2-linked Man to trimannosyl GPI precursors in C . albicans and is necessary for viability . Because addition of a fourth Man to GPIs is of less relative importance in mammals, Smp3p is a potential antifungal target.

Mol Microbiol, 2004 Oct, 54(2), 507 - 19
Candida albicans lacking the frataxin homologue: a relevant yeast model for studying the role of frataxin; Santos R et al.; We cloned the CaYFH1 gene that encodes the yeast frataxin homologue in Candida albicans . CaYFH1 was expressed in Deltayfh1 Saccharomyces cerevisiae cells, where it compensated for all the phenotypes tested except for the lack of cytochromes . Double DeltaCayfh1/DeltaCayfh1 mutant had severe defective growth, accumulated iron in their mitochondria, lacked aconitase and succinate dehydrogenase activity and had defective respiration . The reductive, siderophore and haem uptake systems were constitutively induced and the cells excreted flavins, thus behaving like iron-deprived wild-type cells . Mutant cells accumulated reactive oxygen species and were hypersensitive to oxidative stress, but not to iron . Cytochromes were less abundant in mutants than in wild-type cells, but this did not result from defective haem synthesis . The low cytochrome concentration in mutant cells was comparable to that of iron-deprived wild-type cells . Mitochondrial iron was still available for haem synthesis in DeltaCayfh1/DeltaCayfh1 cells, in contrast to S . cerevisaeDeltayfh1 cells . CaYFH1 transcription was strongly induced by iron, which is consistent with a role of CaYfh1 in iron storage . Iron also regulated transcription of CaHEM14 (encoding protoporphyrinogen oxidase) but not that of CaHEM15 (encoding ferrochelatase) . There are thus profound differences between S . cerevisiae and C . albicans in terms of haem synthesis, cytochrome turnover and the role of frataxin in these processes.

AIDS Patient Care STDS, 1998 Aug, 12(8), 625 - 7
The treatment of oropharyngeal candidiasis in HIV-infected patients with oral amphotericin B suspension; Hood S et al.; Oropharyngeal candidiasis (OPC) is the most frequent opportunistic infection associated with HIV infection . Therapies such as topical clotrimazole and nystatin, as well as oral azoles, which had previously been effective prior to the advent of HIV, are increasingly only partially effective in OPC in HIV infection . The effectiveness of oral amphotericin B suspension for OPC is described in 17 HIV-infected patients whose response to other therapies had been unsatisfactory . Three patients yielded isolates of Candida albicans with a minimum inhibitory concentration (MIC) to fluconazole of >/=16 microg/mL . Eleven patients received amphotericin B suspension monotherapy . Of the 17 patients, the symptoms of six resolved entirely, seven patients partially responded, and four failed therapy . These data suggest that amphotericin B suspension may be a useful additional therapy for OPC in HIV-infected patients.

Chem Pharm Bull (Tokyo), 2004 Oct, 52(10), 1235 - 7
A new phenanthrene glycoside and other constituents from Dioscorea opposita; Sautour M et al.; Phytochemical investigation of the rhizome of Dioscorea opposita has led to the isolation of a new phenanthrene glycoside, 3,4,6-trihydroxyphenanthrene-3-O-beta-D-glucopyranoside (1), and five known compounds, soyacerebroside I (2), adenosine (3), beta-sitosterol (4), palmitic acid (5) and palmitoyloleoylphosphatidylcholine (6) . Their structures were determined by spectroscopic methods, including 1D- and 2D-NMR . Compounds 1-6 exhibited no antifungal activity against the human pathogenic yeasts Candida albicans, C . glabrata and C . tropicalis.

Eur J Med Chem, 2004 Oct, 39(10), 827 - 34
Esters, amides and substituted derivatives of cinnamic acid: synthesis, antimicrobial activity and QSAR investigations; Narasimhan B et al.; A series of esters (I(a-k)), substituted derivatives (II(a-d)) and amides (III(a-q)) of cinnamic acid were synthesized and evaluated as antibacterial and antifungal agents . All the derivatives belonging to the series I, II and III showed antimicrobial activity comparable to the standard . Compounds I(f) and II(c) proved to be the most effective compounds . Quantitative structure-activity relationship (QSAR) investigation with multiple linear regression analysis was applied to find a correlation between different calculated physicochemical parameters of the compounds and biological activity . The quantitative models relating the structural features of cinnamic acid derivatives I(a-k), II(a-d) and III(a-q) and their antimicrobial activity showed that Gram negative Escherichia coli and Candida albicans (fungus) were the most sensitive microorganisms.

BMC Mol Biol . 2004 Oct 04;5(1):17.
Polyadenylation of ribosomal RNA by Candida albicans also involves the small subunit; Fleischmann J et al.; BACKGROUND: Candida albicans is a polymorphic fungus causing serious infections in immunocompromised patients . It is capable of shifting from yeast to germinating forms such as hypha and pseudohypha in response to a variety of signals, including mammalian serum . We have previously shown that some of the large 25S components of ribosomal RNA in Candida albicans get polyadenylated, and this process is transiently intensified shortly after serum exposure just prior to the appearance of germination changes . RESULTS: We now present data that this process also involves the small 18S subunit of ribosomal RNA in this organism . Unlike the large 25S subunit, polyadenylation sites near the 3' end are more variable and no polyadenylation was found at the reported maturation site of 18S . Similar to 25S, one or more polyadenylated mature sized 18S molecules get intensified transiently by serum just prior to the appearance of hypha . CONCLUSIONS: The transient increase in polyadenylation of both the large and the small subunits of ribosomal RNA just prior to the appearance of hypha, raises the possibility of a role in this process.

Dermatology, 2004, 209(3), 190 - 6
Prevalence and risk factors for superficial fungal infections among Italian Navy Cadets; Ingordo V et al.; BACKGROUND: Limited studies on the prevalence and risk factors for superficial mycoses are available . OBJECTIVE: The aim of this paper was to evaluate the prevalence and risk factors for superficial mycoses (dermatophytes and Candida spp.) in a sample of young Italian people resident at a military school . METHODS: A total of 1,024 young cadets from the Italian Navy Petty Officers School in Taranto, including 975 (95.21%) males and 49 (4.79%) females, mean age 22.5 +/- 3.0 years (range 18-30), were consecutively examined by the same observer . A complete dermatological examination was performed on all the subjects, and skin scrapings for microscopy and fungal culture were obtained from suspected lesions . All the subjects completed a questionnaire providing information on sports practice, swimming-pool attendance, marching, wearing shower sandals, frequent use of 'gummed' shoes, history of severe traumas to the nails, presence of hyperhidrosis and history of superficial mycoses . The affected subjects were also asked if they were aware of their condition . Data were analysed by the Statistical Analysis System, version 8.0 . The Fisher exact test and odds ratios were calculated . RESULTS: A total of 33 subjects (3.2%) were found to suffer from a mycologically confirmed fungal infection (3% by dermatophytes and 0.2% by Candida albicans): tinea pedis/Candida intertrigo of the feet was suspected in 126 (12.1%) subjects and confirmed in 30 (2.9%), including 28 cases of tinea pedis and 2 cases of Candida intertrigo; tinea cruris/Candida intertrigo of the groin was suspected in 28 (2.7%) subjects, but confirmed in only 1 case (0.1%); onychomycosis was suspected in 64 (6.1%) subjects and confirmed in 2 cases (0.2%) . The organism most frequently responsible in tinea pedis was Trichophyton mentagrophytes var . interdigitale (82.1%) . The same species (50%) and T . mentagrophytes var . mentagrophytes (50%) were associated with tinea unguium, Epidermophyton floccosum was the only species detected in tinea cruris . Non-dermatophytic filamentous fungi (Penicillium spp., Fusarium spp., Aspergillus spp . and Paecilomyces spp.), not considered pathogenic, were isolated in 48 samples . None of the risk factors analysed were significantly associated with fungal infection . Only 2 subjects out of the 33 people affected were aware of their condition . They both had tinea pedis . CONCLUSION: The prevalence of mycoses in sailors living in an Italian military school was lower than rates detected in other military populations . This may be due to the cadets' lifestyle and environmental conditions . The most frequent infection was tinea pedis, mainly caused by T . interdigitale . None of the investigated risk factors were significantly associated with the disease, and most of the affected individuals were not aware of their condition.

Rev Iberoam Micol, 2004 Mar, 21(1), 29 - 34
{Effect of metabolic substances of oral Actinomyces on Candida albicans}; Gutierrez de Annan S et al.; Actinomyces naeslundii, Actinomyces viscosus and Candida albicans are associated with root cavity . The aim of this study was to determine, in vitro, the effect produced by the metabolic substances elaborated by Actinomyces naeslundii and Actinomyces viscosus on Candida albicans . The strains were isolated of saliva . There were used the double plaque diffusion method (DPDM) and the method of radial diffusion (MRD) . The effect of the time of incubation and of different concentrations of metabolic substances elaborated by Actinomyces naeslundii and Actinomyces viscosus on the kinetics of growth of C . albicans were studied . Later, the nature of the substances produced by the two strains of Actinomyces was determined . It was found that there was no inhibition of the growth of C . albicans by A . naeslundii and A . viscosus in the DPDM and the MRD . There was stimulation of the growth of C . albicans by the two strains of Actinomyces when the DPDM was used . In the MRD the results were negative . Metabolic substances produced by both species stimulated the growth of C . albicans in low concentrations but at high concentrations inhibition was observed . The best concentration of the stimulating factor, a protein substance stable to 70 degrees C, corresponds to a dilution of 1/80 . The inhibition of the growth of C . albicans was produced by the decrease of the pH, the higher effect being obtained with the dilution 1/5 . The metabolic substances produced by A . naeslundii and A . viscosus can have both inhibitory and stimulant effects on C . albicans, according to their concentration . These metabolic interactions would condition the proportion of C . albicans in the oral microbial ecosystems.

Plast Reconstr Surg, 2004 Oct, 114(5), 1170 - 8
Fungal growth inside saline-filled implants and the role of injection ports in fungal translocation: in vitro study; Saray A et al.; Infection is a serious complication of breast augmentation and tissue expansion with inflatable devices . Several reports have shown that fungi may be able to survive, colonize, and even cause infection in saline-filled devices . The mechanism of how they penetrate, spread, and colonize inside the inflatable implants is not exactly understood . The authors assessed both the expander membrane and the port in terms of leakage and penetration of Candida albicans and Aspergillus niger in an in vitro model . Thirty saline-filled expanders connected to the injection port were placed in sterile containers filled with tryptic soy broth culture medium to simulate the clinical situation in phases I and II . Intact and multipunctured ports were used in the first and second phases of the study, respectively . Either the container or the implant was inoculated with one of these fungi, and six implants in containers without fungal inoculation served as controls . As a third phase, intraluminal survival of fungi was investigated in saline-filled containers (n = 12) in 21 days . The silicone membrane, with its intact connecting tube and port, was impermeable to these fungi, whereas both fungi were able to diffuse inside-out or outside-in through the punctured ports . C . albicans did not survive beyond 18 days in saline, whereas A . niger continued to multiply at day 21 . Chemical analyses of the implant fluids revealed that the contents of the culture medium diffused into the implants in phases I and II . The data show that an intact silicone membrane is impermeable to fungi, and punctured ports allow translocation of fungi into the implants . Fungi can grow and reproduce in a saline-only environment, and their survival periods differ among the species . Furthermore, their survival may be enhanced by the influx of substances through the implant shell.

J Antimicrob Chemother, 2004 Nov, 54(5), 909 - 14 Epub 2004 Sep 29.
Evaluation of carvacrol and eugenol as prophylaxis and treatment of vaginal candidiasis in an immunosuppressed rat model; Chami F et al.; OBJECTIVES: Anticandidal activity of carvacrol and eugenol, the major phenolic components of oregano and clove essential oils, respectively, were tested in vivo . METHODS: Efficacy evaluation of carvacrol and eugenol in the prophylaxis and treatment of experimental vaginal candidiasis was performed in immunosuppressed rats . The anticandidal activity was analysed by microbiological and histological techniques and was compared with that of nystatin . RESULTS: Microbiologically, prophylactic treatment with carvacrol eradicated the vaginal fungal burden of infected rats, whereas eugenol reduced the number of colony counts of Candida albicans in vaginas of infected rats by 98.9% 10 days after inoculation . Therapeutic treatment for 7 consecutive days with carvacrol was able to eradicate the vaginal candidal burden in 7/9 of the infected rats and reduced the number of colony counts of C . albicans in vaginas of the two remaining rats by 98% . Treatment with eugenol completely cured 2/9 of the infected animals, but the 7/9 still infected showed an 84% reduction of colony counts of C . albicans in their vaginas . Histologically, in all treated rats, no Candida organisms were found in the lumina of the vagina; this was in contrast to control groups in which many yeasts, strongly stained with periodic acid-Schiff, were observed . The results obtained with nystatin used at 10-fold minimal inhibitory concentration confirm the validity of this model . CONCLUSIONS: Carvacrol and eugenol could be considered as promising products in the treatment of vaginal candidiasis . This work is a preliminary contribution to the development of a new generation of efficient and natural antifungal agents for curative treatment and prophylaxis.

Rev Iberoam Micol, 2003 Jun, 20(2), 52 - 4
Adherence of Candida albicans and Candida dubliniensis to buccal and vaginal cells; Vidotto V et al.; Twenty-seven Candida albicans strains and 26 Candida dubliniensis strains, isolated from HIV patients, were tested for their adherence to buccal and vaginal epithelial cells . Both species showed important levels of adhesion to buccal and vaginal epithelial cells, although C . albicans showed the highest levels of adhesion . These results suggest that both Candida species are well adapted, in terms of adhesion capability, to the oral and vaginal environment.

Heart Surg Forum, 2004 Jul 01, 7(4), E312 - 4
Candida albicans endocarditis and a review of fungal endocarditis: case report; Filizcan U et al.; Endocarditis due to fungal etiology is rare, but it is the most severe form of infective endocarditis . Fungal endocarditis is commonly complicated by systemic embolizations, and the difficulty in isolating the fungi with routine blood cultures complicates the diagnostic process . In these culture-negative cases of endocarditis, etiologic diagnosis is made with histopathologic examination of the cardiac valve, embolic materials, and systemic ulcers . In this case report, the presented patient with fungal endocarditis and its neurologic complications was treated with a surgical and medical approach.

Free Radic Res, 2004 Jul, 38(7), 739 - 50
A comparison of the effects of ocular preservatives on mammalian and microbial ATP and glutathione levels; Ingram PR et al.; The aim of this study was to investigate the mechanism of action of the preservative sodium chlorite (NaClO2), and the relationship with intracellular glutathione depletion . A detailed comparison of the dose responses of two cultured ocular epithelial cell types and four species of microorganism was carried out, and comparisons were also made with the quaternary ammonium compound benzalkonium chloride (BAK), and the oxidant hydrogen peroxide (H2O2) . The viability of mammalian and microbial cells was assessed in the same way, by the measurement of intracellular ATP using a bioluminescence method . Intracellular total glutathione was measured by reaction with 5,5'-dithiobis-2-nitrobenzoic acid in a glutathione reductase-dependent recycling assay . BAK and H2O2 caused complete toxicity to conjunctival and corneal epithelial cells at approximately 25 ppm, in contrast to NaClO2, where > 100 ppm was required . The fungi Candida albicans and Alternaria alternata had a higher resistance to NaClO2 than the bacteria Staphyloccus aureus and Pseudomonas aeruginosa, but the bacteria were extremely resistant to H2O2 . NaClO2 caused substantial depletion of intracellular glutathione in all cell types, at concentrations ranging from < 10 ppm in Pseudomonas, 25-100 ppm in epithelial cells, to > 500 ppm in fungal cells . The mechanisms of cytotoxicity of NaClO2, H2O2 and BAK all appeared to differ . NaClO2 was found to have the best balance of high antibacterial toxicity with low ocular toxicity . The lower toxicity of NaClO2 to the ocular cells, compared with BAK and H2O2, is in agreement with fewer reported adverse effects of application in the eye.

Phytochemistry, 2004 Sep, 65(18), 2569 - 75
Azaphilone pigments from a yellow mutant of the fungus Monascus kaoliang; Jongrungruangchok S et al.; Azaphilone pigments, monascusones A (1) and B (2), together with two known azaphilones, monascin (3) and FK17-P2b2 (4), were isolated from the CH2Cl2 extract of a yellow mutant of the fungus M . kaoliang grown on rice . Structures of the isolated compounds were elucidated by analyses of spectroscopic data . Monascusone A (1), the major metabolite of M . kaoliang, showed no antimalarial (against Plasmodium falciparum), antitubercular (against Mycobacterium tuberculosis H37Ra), and antifungal (toward Candida albicans) activities . Compound 1 exhibited no cytotoxicity against BC (breast cancer) and KB (human epidermoid carcinoma of cavity) cell lines.

Infect Genet Evol, 2004 Sep, 4(3), 243 - 52
Multilocus sequence typing of Candida albicans: strategies, data exchange and applications; Bougnoux ME et al.; Multilocus sequence typing of Candida albicans: strategies, data exchange and applications . Bougnoux, M.-E., Aanensen, D.M., Morand, S., Theraud, M., Spratt, B.G., and d'Enfert, C . Infection, Genetics and Evolution . C . albicans is a commensal of humans and animals but is also the main fungal pathogen of humans, ranking fourth among the microorganisms responsible for hospital-acquired bloodstream infections . Information on the genetic diversity and dynamics of the C . albicans population and on the characteristics of C . albicans strains causing invasive infections in immunocompromised patients is important in order to adapt prevention policies . Important results in this field have been obtained using the Ca3 fingerprinting probe . Recently, multilocus sequence typing (MLST) based on the sequencing of 6-8 selected house-keeping genes and identification of polymorphic nucleotide sites has been introduced for the characterization of C . albicans isolates . Combination of the alleles at the different loci results in unique diploid sequence types (DSTs) that can be used to discriminate strains . MLST has now been successfully applied to study the epidemiology of C . albicans in the hospital as well as the diversity of C . albicans isolates obtained from diverse ecological niches including human and animal hosts . Furthermore, MLST data for C . albicans are available in a public database that provides a new resource to evaluate the worldwide diversity of C . albicans and the relationships of isolates identified at various locations.

Rev Med Chil, 2004 Feb, 132(2), 151 - 9
{Antimicrobial activity of actinomycetes isolated from aquatic environments in southern Chile}; Leiva S et al.; BACKGROUND: The easy access and inappropriate use of antimicrobials led to selection and spread of resistant microorganisms strains . It is imperative to search for new and more effective antimicrobials . One strategy is the screening of metabolites produced by microorganisms found in the environment . Actinomycetes are a potential source of new drugs . AIM: To isolate actinomycetes from sediments of Chilean rivers and lakes and to screen them for antimicrobial activity against reference bacterial strains and pathogenic fungi . MATERIAL AND METHODS: Actinomycetes were isolated from sediment samples, using casein-starch agar . The antimicrobial activity against 3 bacterial species and 7 fungal species was tested using the disc diffusion method . For the extraction of active metabolites, culture broths of antagonistic actinomycetes were extracted with organic solvents followed by testing the antibiotic activity . RESULTS: A total of 62 strains of actinomycetes were isolated, mainly Streptomyces sp (83.9%) . Thirty six strains (58.1%) showed antimicrobial activity, mainly against Bacillus subtilis and Candida albicans . Some isolates inhibited a wide spectrum of indicator strains, like LRI 4A strain (Streptomyces sp) that inhibited Bacillus subtilis, Candida albicans and 4 filamentous fungi . CONCLUSIONS: Lakes and rivers of Southern Chile are an important reservoir of antagonistic actinomycetes, a potential source of new drugs.

Yeast, 2004 Sep, 21(12), 1025 - 33
Tandem affinity purification of the Candida albicans septin protein complex; Kaneko A et al.; A novel vector was constructed to enable the integrative marking of individual genes and the affinity purification of interacting molecules within protein complexes from Candida albicans using a tandem 6 x histidine and FLAG epitope tag . The system was verified by purifying the C . albicans septin complex (a self-associating complex of cytoskeletal proteins) from both yeast and hyphal cells . One-step affinity purification was insufficient for purification of the protein complex, whereas tandem affinity purification (TAP) gave an extensively purified protein complex with a very low background . Electrophoretic and mass spectrometry analysis showed that the affinity-purified C . albicans septin complex, which comprises predominantly CaCdc3p, CaCdc10p, CaCdc11p, CaCdc12p and CaSep7p, was not affected by cell morphology . The purified septin complex appeared to have a stoichiometry of 2 CaCdc3p, 1-2 CaCdc10p, 1 CaCdc11p, 2 CaCdc12p and < or = 1 CaSep7p . The successful application of TAP to the purification and analysis of the C . albicans septin complex indicates that this technology will have much wider application to proteomic studies of this pathogenic fungus . Copyright (c) 2004 John Wiley & Sons, Ltd.

Braz J Med Biol Res, 2004 Oct, 37(10), 1455 - 61 Epub 2004 Sep 22.
Respiration, oxidative phosphorylation, and uncoupling protein in Candida albicans; Cavalheiro RA et al.; The respiration, membrane potential (Deltapsi), and oxidative phosphorylation of mitochondria in situ were determined in spheroplasts obtained from Candida albicans control strain ATCC 90028 by lyticase treatment . Mitochondria in situ were able to phosphorylate externally added ADP (200 microM) in the presence of 0.05% BSA . Mitochondria in situ generated and sustained stable mitochondrial Deltapsi respiring on 5 mM NAD-linked substrates, 5 mM succinate, or 100 microM N,N,N',N'-tetramethyl-p-phenylenediamine dihydrochloride plus 1 mM ascorbate . Rotenone (4 microM) inhibited respiration by 30% and 2 micro M antimycin A or myxothiazole and 1 mM cyanide inhibited it by 85% . Cyanide-insensitive respiration was partially blocked by 2 mM benzohydroxamic acid, suggesting the presence of an alternative oxidase . Candida albicans mitochondria in situ presented a carboxyatractyloside-insensitive increase of Deltapsi induced by 5 mM ATP and 0.5% BSA, and Deltapsi decrease induced by 10 microM linoleic acid, both suggesting the existence of an uncoupling protein . The presence of this protein was subsequently confirmed by immunodetection and respiration experiments with isolated mitochondria . In conclusion, Candida albicans ATCC 90028 possesses an alternative electron transfer chain and alternative oxidase, both absent in animal cells . These pathways can be exceptional targets for the design of new chemotherapeutic agents . Blockage of these respiratory pathways together with inhibition of the uncoupling protein (another potential target for drug design) could lead to increased production of reactive oxygen species, dysfunction of Candida mitochondria, and possibly to oxidative cell death.

Cornea, 2004 Nov, 23(8 Suppl), S36 - 41
Potential use of (1,3)-beta-D-glucan as target of diagnosis and treatment of keratomycosis; Kaji Y et al.; PURPOSE: Major problems in the management of keratomycosis stem from the difficulty of its diagnosis and limited choice of antifungal agents . In the present paper we propose a new method of detecting (1,3)-beta-D-glucan, one of the major components of fungal cell wall, in tears from an animal model of keratomycosis . In addition, we investigated the efficacy of topical application of micafungin, a new antifungal agent that inhibits the activity of (1,3)-beta-D-glucan synthase in this animal model . METHOD: Candida albicans (5 x 10(5) organisms) was inoculated into the corneal stroma of 20 New Zealand White rabbits . The animals were randomly assigned to two groups and treated with subconjunctival injection of 0.5 mL of saline or 0.1% micafungin every day for 3 weeks . The clinical course of keratomycosis in both groups was compared . Before and 3 weeks after the injection of saline or micafungin, 5 microL of tears in each eye were collected by capillary tube . The concentration of (1,3)-beta-D-glucan was quantitatively measured by modified Limulus test . RESULTS: The concentration of (1,3)-beta-D-glucan was significantly higher in keratomycosis model animals than in controls (mean +/- SD, 17.4 +/- 9.4 pg/mL and 2.8 +/- 1.8 pg/mL, respectively) at 21 days after treatment . Subconjunctival injection of micafungin had no significant effect on ocular lesions of keratomycosis until 9 days, after which ocular lesions significantly improved . Subconjunctival application of micafungin decreased the concentration of (1,3)-beta-D-glucan in tears to 4.9 +/- 3.0 pg/mL at 21 days after treatment . CONCLUSIONS: Increased levels of (1,3)-beta-D-glucan in tears were detected in this model of keratomycosis . Measuring the concentration of (1,3)-beta-D-glucan in tears may be a reliable noninvasive method for the diagnosis of keratomycosis . Topical application of micafungin was effective in the treatment of keratomycosis.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 4056 - 8
Sequential therapy with caspofungin and fluconazole for Candida albicans infection; Barchiesi F et al.; A sequential therapy of caspofungin (CAS) and fluconazole (FLC) administration for treatment of Candida albicans infection was investigated . Treatment with CAS followed by FLC was as effective as CAS treatment given alone for the same duration . Our data suggest that switching from CAS to FLC is a potentially explorable therapeutic option for treatment of systemic candidiasis.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3959 - 67
Efficacy of PLD-118, a novel inhibitor of candida isoleucyl-tRNA synthetase, against experimental oropharyngeal and esophageal candidiasis caused by fluconazole-resistant C . albicans; Petraitis V et al.; PLD-118, formerly BAY 10-8888, is a synthetic antifungal derivative of the naturally occurring beta-amino acid cispentacin . We studied the activity of PLD-118 in escalating dosages against experimental oropharyngeal and esophageal candidiasis (OPEC) caused by fluconazole (FLC)-resistant Candida albicans in immunocompromised rabbits . Infection was established by fluconazole-resistant (MIC > 64 microg/ml) clinical isolates from patients with refractory esophageal candidiasis . Antifungal therapy was administered for 7 days . Study groups consisted of untreated controls; animals receiving PLD-118 at 4, 10, 25, or 50 mg/kg of body weight/day via intravenous (i.v.) twice daily (BID) injections; animals receiving FLC at 2 mg/kg/day via i.v . BID injections; and animals receiving desoxycholate amphotericin B (DAMB) i.v . at 0.5 mg/kg/day . PLD-118- and DAMB-treated animals showed a significant dosage-dependent clearance of C . albicans from the tongue, oropharynx, and esophagus in comparison to untreated controls (P </= 0.05, P </= 0.01, P </= 0.001, respectively), while FLC had no significant activity . PLD-118 demonstrated nonlinear plasma pharmacokinetics across the investigated dosage range, as was evident from a dose-dependent increase in plasma clearance and a dose-dependent decrease in the area under the plasma concentration-time curve . The biochemical safety profile was similar to that of FLC . In summary, PLD-118 demonstrated dosage-dependent antifungal activity and nonlinear plasma pharmacokinetics in treatment of experimental FLC-resistant oropharyngeal and esophageal candidiasis.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3845 - 9
Caspofungin uptake is mediated by a high-affinity transporter in Candida albicans; Paderu P et al.; The uptake of the echinocandin drug caspofungin acetate in Candida albicans was evaluated at drug levels at or near the MIC for the organism . Maximal uptake was achieved in 10 min and was energy independent . A saturable transport system, consistent with a facilitated-diffusion carrier, was observed with the unlabeled drug competing with the labeled drug for uptake and efflux . More than 90% of the transported drug was observed in a single kinetic compartment that was available for efflux, indicating that the drug was free in the cytoplasm following uptake . Efflux was also energy independent but was sensitive to the presence of a fully loaded carrier on both faces of the bilayer . Overall, the data presented are consistent with the presence of a high-affinity facilitated-diffusion transporter that mediates caspofungin uptake and could be a potential source of transport-related reduced susceptibility.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3828 - 33
Antimicrobial activity of euplotin C, the sesquiterpene taxonomic marker from the marine ciliate Euplotes crassus; Savoia D et al.; Strains of the marine ciliate protist Euplotes crassus produce exclusive terpenoids called euplotins that play an ecological role . Among these derivatives, euplotin C is the main of four secondary metabolites isolated from cultures of this protozoon and represents the sesquiterpene taxonomic marker from E . crassus . Because different terpenoid metabolites of plant origin showed a certain antimicrobial activity, we assessed the compound euplotin C, purified by high-pressure liquid chromatography and solubilized in two solubility enhancers, against the protozoa Leishmania major and Leishmani infantum, the fungus Candida albicans, and nine strains of gram-positive and gram-negative microorganisms . An activity of euplotin C against Leishmania promastigotes was demonstrated (50% lethal doses were 4.6 or 8.1 microg/ml depending on the agent used to solubilize the compound), while the effect was less evident on Candida and nearly absent on bacteria . A nonsignificant cytotoxicity (50% lethal dose, >200 microg/ml) against the J774 cell line was observed . A leishmanicidal activity was also shown by the living, euplotin-producing cells of E . crassus cultured together with promastigotes; this activity increased with time from 10 min to 6 h of incubation . This study provides an initial rationale for the evaluation of euplotin C and other similar natural products as alternative or possibly synergistic compounds for current antiprotozoon chemotherapeutics.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3690 - 6
Posaconazole is a potent inhibitor of sterol 14alpha-demethylation in yeasts and molds; Munayyer HK et al.; Posaconazole (POS; SCH 56592) is a novel triazole that is active against a wide variety of fungi, including fluconazole-resistant Candida albicans isolates and fungi that are inherently less susceptible to approved azoles, such as Candida glabrata . In this study, we compared the effects of POS, itraconazole (ITZ), fluconazole (FLZ), and voriconazole (VOR) on sterol biosynthesis in strains of C . albicans (both azole-sensitive and azole-resistant strains), C . glabrata, Aspergillus fumigatus, and Aspergillus flavus . Following exposure to azoles, nonsaponifiable sterols were extracted and resolved by liquid chromatography and sterol identity was confirmed by mass spectroscopy . Ergosterol was the major sterol in all but one of the strains; C . glabrata strain C110 synthesized an unusual sterol in place of ergosterol . Exposure to POS led to a decrease in the total sterol content of all the strains tested . The decrease was accompanied by the accumulation of 14alpha-methylated sterols, supporting the contention that POS inhibits the cytochrome P450 14alpha-demethylase enzyme . The degree of sterol inhibition was dependent on both dose and the susceptibility of the strain tested . POS retained activity against C . albicans isolates with mutated forms of the 14alpha-demethylase that rendered these strains resistant to FLZ, ITZ, and VOR . In addition, POS was a more potent inhibitor of sterol synthesis in A . fumigatus and A . flavus than either ITZ or VOR.

Mol Microbiol, 2004 Sep, 53(5), 1451 - 69
Regulatory networks affected by iron availability in Candida albicans; Lan CY et al.; Iron, an essential element for almost every organism, serves as a regulatory signal for the expression of virulence determinants in many prokaryotic and eukaryotic pathogens . Using a custom Affymetrix GeneChip representing the entire Candida albicans genome, we examined the changes in genome-wide gene expression in this opportunistic pathogen as a function of alterations in environmental concentrations of iron . A total of 526 open reading frame (ORF) transcripts are more highly expressed when the levels of available iron are low, while 626 ORF transcripts are more highly expressed in high-iron conditions . The transcripts dominantly affected by iron concentration range from those associated with cell-surface properties to others which affect mitochondrial function, iron transport and virulence-related secreted hydrolases . Moreover gene expression as assayed in DNA microarrays confirms and extends reports of alterations in cell-surface antigens and drug sensitivity correlated with iron availability . To understand how these genes and pathways might be regulated, we isolated a gene designated SFU1 that encodes a homologue of the Ustilago maydis URBS1, a transcriptional repressor of siderophore uptake/biosynthesis . Comparisons between wild-type and SFU1-null mutant strains revealed 139 potential target genes of Sfu1p; many of which are iron-responsive . Together, these results not only expand our understanding of global iron regulation in C . albicans, but also provide insights into the potential role of iron availability in C . albicans virulence .

Org Lett, 2004 Sep 30, 6(20), 3533 - 6
Total synthesis of basiliskamides A and B; Lipomi DJ et al.; {structure: see text} The first enantioselective synthesis of the polyketide antibiotics basiliskamides A and B, which exhibit potent in vivo activity against Candida albicans and Aspergillus fumigatus, has been achieved . Serial asymmetric crotylsilane and crotylboronate additions established the C7-C10 stereochemical tetrad . Takai iodoolefination and palladium-mediated cross-coupling were used to install the (Z,E)-vinyl acrylamide . Spectroscopic data is consistent with the assignment of the absolute configurations of the natural products as (7S,8S,9R,10S).

Presse Med, 2004 Jul 31, 33(13), 866 - 8
{Kidney transplant and Candida albicans arteritis . The importance of analysing the transplant conservation liquid}; Gari-Toussaint M et al.; INTRODUCTION: Candida arteritis can compromise the functional prognosis of the graft or even the life of the transplant recipient . The infection can be transmitted by the graft . OBSERVATION: A 46 year-old woman contracted a Candida albicans ateritis of the graft following a kidney transplant that led to a detransplantation . The yeast was probably transmitted by the graft from the donor, source of an unknown candida infection: it was found in the conservation liquid of the graft itself, and in the renal artery and vascular pedicle . Analysis of of these three elements by enzymatic electrophoresis showed that they were identical . COMMENTARIES: This case report underlines the need to establish guidelines and sanitary safety measures, notably that of systematically placing in culture the concervation solutions and alerting the transplant team if any fungi are isolated.

Infect Immun, 2004 Oct, 72(10), 5868 - 76
Dysregulated inflammatory response to Candida albicans in a C5-deficient mouse strain; Mullick A et al.; Experimental infection of inbred mouse strains with Candida albicans provides a good model system to identify host genetic determinants that regulate onset of, response to, and ultimate outcome of disseminated candidiasis . The A/J mouse strain is exquisitely sensitive to infection with C . albicans, while the C57BL/6J strain is relatively resistant, as measured by survival following intravenous injection of Candida blastospores . This differential susceptibility is caused by an A/J-specific loss-of-function mutation in the C5 component of the complement pathway . C5 plays several critical roles in host response to infection, including target lysis and phagocyte recruitment . Therefore, to determine which of its functions were required for host resistance to candidiasis, a detailed comparative analysis of pathophysiology and host response to acute C . albicans infection was conducted in A/J and C57BL/6J mice . C5-sufficient C57BL/6J mice were found to succumb late in infection due to severe kidney pathology, typified by fungal replication and robust neutrophil-based inflammatory response associated with extensive tissue damage . In contrast, A/J mice were moribund within 24 h postinfection but displayed little if any kidney damage despite an inability to mobilize granulocytes and a high fungal load in the kidney . Rather, C5 deficiency in A/J mice was associated with higher levels of circulating cytokines tumor necrosis factor alpha, interleukin-6, monocyte chemotactic protein 1 (MCP-1), MCP-5, and eotaxin in response to C . albicans . Transfer of the C5-defective allele from A/J onto a C57BL/6J genetic background in recombinant congenic strain BcA17 recapitulated the phenotypic aspects of the susceptibility of A/J mice to C . albicans, confirming the causative role of C5 deficiency in the dysregulated cytokine response.

Adv Perit Dial, 2004, 20, 58 - 61
Successful treatment of Candida infections in peritoneal dialysis patients: case reports and review of the literature; Kleinpeter MA; Infections with Candida species have been associated with significant morbidity and mortality in peritoneal dialysis (PD) patients . Such infections include peritonitis and exit-site infections attributable to Candida species, disseminated candidiasis in immunocompromised patients, and Candida esophagitis . In peritonitis and exit-site infections, both success and failure have been reported with commercially available medications . In disseminated candidiasis and Candida esophagitis, successful treatment and patient recovery depend on the overall nutritional and immune state of the patient . One case each of peritonitis and exit-site infection with non Candida albicans species were successfully treated with oral voriconazole . No literature currently exists on the use of this new product in dialysis patients . Presented here is a treatment strategy that resulted in maintenance of PD in the home setting and catheter survival following completion of treatment . A review of the English-language literature shows mixed outcomes associated with continuation of PD during treatment for Candida infection in PD patients . In conclusion, a commercially available product can be used to successfully treat PD patients who have Candida infections and to maintain the PD catheter for PD.

Microbiol Immunol, 2004, 48(9), 665 - 7
The anthracycline antitumor agents doxorubicin and daunorubicin reduce the activity of Candida albicans phospholipase B; Arai R et al.; A pathogenic yeast, Candida albicans, causes life-threatening infection in immunocompromised patients . Inhibiting the production or release of phospholipase B by C . albicans should reduce direct host cell damage, and inhibit the release of eicosanoids from cells of this microorganism . Of the antitumor agents tested, doxorubicin and daunorubicin inhibited the activity of phospholipase B, and prostaglandin production by C . albicans . These two agents have the potential to inhibit the activity of C . albicans phospholipase B, although the inhibitory concentrations exceeded the clinical dose.

FEMS Yeast Res, 2004 Oct, 5(1), 63 - 72
ABC multidrug transporter Cdr1p of Candida albicans has divergent nucleotide-binding domains which display functional asymmetry; Jha S et al.; In order to ascertain the molecular basis of ATP-mediated drug extrusion by Cdr1p, a multidrug transporter of Candida albicans, we recently have reported that the Walker A motif of the N-terminal nucleotide biding domain (NBD) of this protein contains an uncommon cysteine residue (C193; GXXGXGCS/T) which is indispensable for ATP hydrolysis . This residue is exceptionally conserved in N-terminal NBDs of fungal ABC transporters and hence makes these transporters an evolutionarily divergent group . However, the presence of a conventional lysine residue at a similar position in the Walker A motif of the C-terminal NBD warrants the individual contribution of both the NBDs in the ATP-driven efflux function of such transporters . In this study we have investigated the contribution of this divergent Walker A motif in the context of the full Cdr1p protein under in vivo conditions by swapping these two crucial amino acids (C193K in Walker A motif of N-terminal NBD and K901C in Walker A motif of C-terminal NBD) between the two NBDs . Both the native and the mutant variants of Cdr1p were integrated at the PDR5 locus as GFP-tagged fusion proteins and were hyper-expressed . Our study shows that both C193K- and K901C-expressing cells elicit a severe impairment of Cdr1p's ATPase function . However, both these mutations have distinct phenotypes with respect to other functional parameters such as substrate efflux and drug resistance profiles . In contrast to C193K, K901C mutant cells were substantially hypersensitive to the tested drugs (fluconazole, ansiomycin, miconazole and cycloheximide) and were unable to expel rhodamine 6G . Our results for the first time show that both NBDs influence the Cdr1p function asymmetrically, and that the positioning of the cysteine and lysine residues within the respective Walker A motifs is functionally not interchangeable.

Diagn Microbiol Infect Dis, 2004 Sep, 50(1), 25 - 32
Evaluation of fluconazole resistance mechanisms in candida albicans clinical isolates from HIV-infected patients in Brazil; Goldman GH et al.; In this study, we describe resistance mechanisms in fluconazole-resistant isolates of C . albicans isolated from AIDS patients from nine Brazilian hospitals . These mechanisms include the presence of point mutations in the ERG11 gene and overexpression of ERG11, and several genes encoding efflux pumps, as measured by quantitative real-time reverse transcriptase polymerase chain reaction . Several fluconazole-resistant strains had multiple mechanisms of resistance . Four mutations previously described, Y132F, K143R, E266D, and V437I, were identified among the strains, whereas some isolates contained more than one mutation . Fourteen novel mutations were identified . Interestingly, all Brazilian fluconazole-resistant isolates showed homozygosity at mating-type loci (MTL) associated with fluconazole resistance . This is the first comprehensive assessment at molecular level of mechanisms of fluconazole resistance in C . albicans isolates from South America.

Int J Antimicrob Agents, 2004 Oct, 24(4), 401 - 4
Serotonin (5-HT) enhances the activity of amphotericin B against Aspergillus fumigatus in vitro; Heller I et al.; We investigated the in vitro synergistic antifungal potential of combining serotonin (5-HT) and sertraline with amphotericin B and itraconazole against clinical isolates of Aspergillus spp . Synergy tests were performed using the chequerboard microdilution method . Activity was measured against Aspergillus fumigatus (n = 7), Aspergillus flavus (n = 3) and Aspergillus terreus (n = 2), and compared with that for Candida albicans and Candida parapsilosis . The fractional inhibitory concentration (FIC) indices ranged between 0.25 and 3 for the various isolates tested . 5-HT was shown to enhance the activity of amphotericin B against Aspergillus spp . Combination studies with 5-HT and itraconazole and with sertraline and itraconazole or amphothericin B showed different activities for the various strains, including synergism (FIC < 1.0), additivity (FIC = 1), and indifference (FIC between 1.0 and 2.0) . 5-HT and sertraline showed antagonistic activity (FIC > 2) with amphotericin B and itraconazole against C . parapsilosis.

Bioorg Med Chem Lett, 2004 Oct 18, 14(20), 5133 - 7
Quinazolinone fungal efflux pump inhibitors . Part 2: In vitro structure-activity relationships of (N-methyl-piperazinyl)-containing derivatives; Watkins WJ et al.; Structure-activity relationships of a novel series of fungal efflux pump inhibitors with respect to potentiation of the activity of fluconazole against strains of Candida albicans and Candida glabrata over-expressing ABC-type efflux pumps are systematically explored.

Proteomics, 2004 Oct, 4(10), 3084 - 106
Proteomics-based identification of novel Candida albicans antigens for diagnosis of systemic candidiasis in patients with underlying hematological malignancies; Pitarch A et al.; Systemic candidiasis remains a major cause of disease and death, particularly among patients suffering from hematological malignancies . In an attempt to contribute to the discovery of useful biomarkers for its diagnosis and therapeutic monitoring, we embarked on a mapping of Candida albicans immunogenic proteins specifically recognized by antibodies produced during the natural course of systemic Candida infection in this high-risk population . About 85 immunoreactive protein species were detected with systemic candidiasis patients' serum specimens by using immunoproteomics (i.e., two-dimensional electrophoresis followed by Western blotting), and identified through a combination of peptide mass fingerprinting by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS), de novo peptide sequencing using nano-electrospray ionization-ion trap (ESI-IT) MS, and genomic database searches . This proteomic approach has led to the characterization of 42 different housekeeping enzymes as C . albicans antigens . Their biological significance is also discussed . Furthermore, this study is the first to report that 26 of them exhibit antigenic properties in C . albicans, and 35 of them become targets of the human antibody response to systemic candidiasis . Our findings suggest that the production of antibodies to C . albicans phosphoglycerate kinase and alcohol dehydrogenase during systemic candidiasis could be associated with a differentiation of the human immune response . We also highlight the relationship between changes in maintenance of circulating levels of specific anti-Candida antibodies and patients' outcome . Some of these variations, especially the rise of high anti-enolase antibody concentrations, appear to be related to recovery from systemic candidiasis in these patients, which might serve as markers for predicting their outcome . This approach could therefore provide new challenges for diagnosis and clinical follow-up of these fungal infections, and even for antifungal drug or vaccine design.

Proteomics, 2004 Oct, 4(10), 3007 - 20
Low virulent strains of Candida albicans: unravelling the antigens for a future vaccine; Fernandez-Arenas E et al.; Several low virulent Candida albicans mutant strains: CM1613 (deleted in the Mitogen Activated Protein (MAP) Kinase MKC1), CNC13 (deleted in the MAP-kinase HOG1) and the morphological mutant 92' were used as vaccines employing a murine model of systemic candidiasis . In this vaccination trial, only the CNC13 strain was able to induce protection against a subsequent infection with a lethal dose of the wild-type strain . The protection induced by CNC13 vaccinated animals resulted in 60-70% percent of survival . These results demonstrate that collaboration between cellular and humoral responses, induced by the CNC13 mutant, elicited a long lasting and effective protection . Using a proteomic approach (two-dimensional gel electrophoresis followed by Western blotting), twenty-five C . albicans immunogenic proteins were detected and identified by matrix-assisted laser desorption/ionization and/or tandem mass spectrometry . We were able to define an antibody pattern in the sera from the nonvaccinating strains (92' and CM1613), which was different from the profile detected in the sera from surviving animals (vaccinated with the CNC13 mutant) . The utility of this proteomic approach has allowed us to identify antigens that induce protective IgG2a antibody isotype in the sera from vaccinated animals: enolase (Eno1p), pyruvate kinase (Cdc19p), pyruvate decarboxylase (Pdc11p), a component from the 40S ribosomal subunit (Bel1p), triosephosphate isomerase (Tpi1p), DL-glycerol phosphatase (Rhr2p), fructose-bisphosphate aldolase (Fba1p) and two new protective antigens: IMP dehydrogenase (Imh3p), and acetyl-CoA synthetase (Acs2p) . The antigenic proteins that promote protective antibodies described in this work are excellent candidates for a future fungal vaccine; their heterologous expression and vaccine design is currently underway.

Genes Immun, 2004 Nov, 5(7), 576 - 87
Genetic analysis of innate immunity in resistance to Candida albicans; Tuite A et al.; Systemic candidiasis is a significant cause of nosocomial infections and the mechanisms of defense against Candida albicans in humans remain poorly understood . Studies in animal models have demonstrated the importance of innate immunity in controlling the response to infection . Although Th1 cytokines have been shown to direct the overall outcome of infection, the precise role of the Th1/Th2 response and, more generally, the adaptive immune response as a whole, in systemic candidiasis, appears to apply mainly to the development of resistance to reinfection . A genetic approach to the identification of host factors regulating pathogenesis and susceptibility to C . albicans infection has been used in humans and in mouse models of infection . Mouse mutants bearing experimentally induced mutations in specific genes have provided a systematic tool for directly assessing the role of individual proteins in C . albicans susceptibility . Inbred mouse strains have been valuable in showcasing the spectrum of naturally occurring variations in initial susceptibility to infection, and type of disease developed . Crosses between resistant and susceptible strains have led to the detection of additional gene effects affecting innate immunity . Of particular interest is the major effect of a naturally occurring loss-of-function mutation in the C5 complement component that has become fixed in many inbred strains . These and other studies have shown that both a functional complement pathway and robust inflammatory response are critical for resistance to C . albicans.

Mol Biol Cell, 2004 Dec, 15(12), 5528 - 37 Epub 2004 Dec.
Multivesicular body-ESCRT components function in pH response regulation in Saccharomyces cerevisiae and Candida albicans; Xu W et al.; The ESCRT-I, -II, and -III protein complexes function to create multivesicular bodies (MVBs) for sorting of proteins destined for the lysosome or vacuole . Prior studies with Saccharomyces cerevisiae have shown that the ESCRT-III protein Snf7p interacts with the MVB pathway protein Bro1p as well as its homolog Rim20p . Rim20p has no role in MVB formation, but functions in the Rim101p pH-response pathway; Rim20p interacts with transcription factor Rim101p and is required for the activation of Rim101p by C-terminal proteolytic cleavage . We report here that ESCRT-III proteins Snf7p and Vps20p as well as all ESCRT-I and -II proteins are required for Rim101p proteolytic activation in S . cerevisiae . Mutational analysis indicates that the Rim20p N-terminal region interacts with Snf7p, and an insertion in the Rim20p "Bro1 domain" abolishes this interaction, as determined with two-hybrid assays . Disruption of the MVB pathway through mutations affecting non-ESCRT proteins does not impair Rim101p processing . The relationship between the MVB pathway and Rim101p pathway is conserved in Candida albicans, because mutations in four ESCRT subunit genes abolish alkaline pH-induced filamentation, a phenotype previously seen for rim101 and rim20 mutants . The defect is suppressed by expression of C-terminally truncated Rim101-405p, as expected for mutations that block Rim101p proteolytic activation . These results indicate that the ESCRT complexes govern a specific signal transduction pathway and suggest that the MVB pathway may provide a signal that regulates pH-responsive transcription.

J Microbiol Methods, 2004 Nov, 59(2), 293 - 7
Comparative evaluation of four commercial tests for presumptive identification of Candida albicans; Cardenes CD et al.; Four commercially available tests (Albicans ID2, Chromalbicans Agar, CHROMagar Candida, and BactiCard Candida) and the germ tube (GT) test for presumptive identification of Candida albicans were evaluated using clinical isolates of C . albicans (n=89) and of non-albicans yeasts (n=107) . Sensitivities and specificities of all tests regarding the identification of C . albicans were greater than 92%, except for Chromalbicans Agar plates (88.7% after 48 h) and their specificity was 86% . Overall, the four commercial systems were easy to use and are good systems for the routine identification of C . albicans.

J Infect Chemother, 2004 Aug, 10(4), 253 - 5
Protective effects of human saliva on experimental murine oral candidiasis; Kamagata-Kiyoura Y et al.; The protective effects of human saliva, known to contain various proteinaceous factors that have anti- Candida activity in vitro on oral candidiasis in the mouse model, were examined in vivo . Oral candidiasis was established by oral inoculation of viable Candida albicans (C . albicans) cells to ICR mice, 24 h after administration of predonisolone . These mice received orally 0.1 ml human saliva or sterile distilled water into the oral cavity a total of five times at specific intervals: 3, 6, 12, 24, and 36 h after inoculation . Seventy-two hours after inoculation with C . albicans, viable C . albicans cells in the oral cavity of the mice were counted and a subjective score for the extent of white patches on the tongue surface determined . The results showed that viable counts were significantly lower in the human saliva group than in the distilled water group (P < 0.05) . Scores for white patches on the tongue were also significantly lower in the saliva group than in the distilled water group (P < 0.05) . These results suggest that administration of human saliva may inhibit the colonization of the oral cavity by C . albicans in mice and the subsequent onset of oral candidiasis.

J Clin Microbiol, 2004 Sep, 42(9), 4379 - 82
Neonatal Candida albicans septic thrombosis of the portal vein followed by cavernous transformation of the vessel; Pacifico L et al.; We report two premature neonates with Candida albicans septic thrombosis of the portal vein who developed, in very early childhood, the sonographic appearance of cavernous transformation of the vessel and/or clinical signs of extrahepatic portal hypertension.

FEMS Immunol Med Microbiol, 2004 Oct 1, 42(2), 219 - 24
Apoptosis of phagocytic cells induced by Candida albicans and production of IL-10; Gasparoto TH et al.; Macrophages co-incubated with Candida albicans strain CR1 in vitro showed early signs of apoptosis, but evolved to necrosis after 2 h . In this study, we investigated whether strain CR1 caused apoptosis or necrosis of macrophages after its inoculation into mice peritoneal cavity, and whether this correlated with the secretion of IL-10 . Peritoneal macrophages from mice that received an inoculum of C . albicans CR1 showed signs of apoptosis and necrosis from 30 min to 2 h afterwards, whereas heat-killed C . albicans did not cause those effects . IL-10 production was low during the first 6 h post-infection, when macrophages predominated in the peritoneal exudate, whereas its higher production after 24 h correlated with an increase of neutrophils in the exudate . Treatment of CR1 with pepstatin (an inhibitor of proteinases) prevented the process of apoptosis and significantly reduced IL-10 production, suggesting that the increased production of IL-10 was caused by processes occurring during the initial phase of infection, such as apoptosis, necrosis and uptake of death cells.

FEMS Immunol Med Microbiol, 2004 Oct 1, 42(2), 181 - 5
Effects of human lactoferrin on the cytoplasmic membrane of Candida albicans cells related with its candidacidal activity; Viejo-Diaz M et al.; Human lactoferrin is an innate host defence protein with antimicrobial activity that exerts a candidacidal effect in a cation concentration-dependent manner . We investigated the ability of this cationic protein (with an isoelectric point of 8.7) to permeabilize the cytoplasmic membrane of Candida albicans cells . Despite minor K(+)-release in lactoferrin-treated C . albicans cells, the killing effect was not related to an extensive membrane permeabilization, as indicated by: (a) the non-release of macromolecular cytosolic constituents; (b) the non-permeabilization for extracellular propidium iodide nor for intracellular accumulated calcein; and (c) the inability to disrupt the phospholipid bilayer of 8-aminonaphthalene-1,3,6, trisulfonic acid/ {Formula: see text} -xylene-bis-pyridiniumbromide-loaded liposomes . These results suggest that lactoferrin exerts its candidacidal effect through a mechanism different from membrane permeabilization described for other cationic peptides.

FEMS Immunol Med Microbiol, 2004 Oct 1, 42(2), 155 - 66
The solubilization and biological activities of Aspergillus beta-(1 --> 3)-D-glucan; Ishibashi K et al.; We have recently demonstrated that the cell wall beta-glucan of Candida albicans could be solubilized by sodium hypochlorite, followed by dimethylsulfoxide-extraction (NaClO-DMSO method) . In this study, applying this method to Aspergillus spp., we prepared mycelial cell wall beta-glucan and examined its physical properties and immunotoxicological activity . The acetone-dried mycelia of Aspergillus spp . were oxidized by the NaClO-DMSO method . An analysis of (13)C NMR spectra revealed the preparations to be composed of alpha-(1 --> 3) and beta-(1 --> 3)-D-glucan . Also, the proportion of alpha-(1 --> 3) and beta-(1 --> 3)-D-glucan varied . Furthermore, a solubilized Aspergillus beta-glucan (ASBG) was prepared from OX-Asp by urea-autoclave treatment . ASBG showed limulus activity similar to Candida solubilized beta-glucan (CSBG), and there was little difference in the activity of ASBG between various Aspergillus spp . ASBG affected the production of IL-8 by human peripheral blood mononuclear cells (PBMC) . ASBG should be useful for analyzing the clinical role of beta-glucan.

Clin Diagn Lab Immunol, 2004 Sep, 11(5), 977 - 82
Antigen-specific gene expression profiles of peripheral blood mononuclear cells do not reflect those of T-lymphocyte subsets; McLaren PJ et al.; Advances in microarray technology have allowed for the monitoring of thousands of genes simultaneously . This technology is of particular interest to immunologists studying infectious diseases, because it provides tremendous potential for investigating host-pathogen interactions at the level of immune gene expression . To date, many studies have focused either on cell lines, where the physiological relevance is questionable, or on mixed cell populations, where the contributions of individual subpopulations are unknown . In the present study, we perform an intrasubject comparison of antigen-stimulated immune gene expression profiles between a mixed population of peripheral blood mononuclear cells (PBMC) and the two predominant cell types found in PBMC, CD4+ and CD8+ T lymphocytes . We show that the microarray profiles of CD4+ and CD8+ T lymphocytes differ from each other as well as from that of the mixed cell population . The independence of the gene expression profiles of different cell types is demonstrated with a ubiquitous antigen (Candida albicans) as well as with a disease-specific antigen (human immunodeficiency virus p24) . This study has important implications for microarray studies of host immunity and underscores the importance of profiling the expression of specific cell types.

Curr Opin Microbiol, 2004 Aug, 7(4), 350 - 7
Candida morphogenesis and host-pathogen interactions; Whiteway M et al.; The human fungal pathogen Candida albicans has many morphological forms . Recent advances in genomics and cell biology are providing an improved understanding of the molecular regulation of cell shape, and providing insights into the relationships between morphogenesis and virulence . This understanding may improve our ability to develop strategies to combat Candida infections.

Curr Opin Microbiol, 2004 Aug, 7(4), 342 - 9
Candida albicans cell wall glycans, host receptors and responses: elements for a decisive crosstalk; Poulain D et al.; Candida albicans has adapted to live on the mucosal surfaces of animals . The human species has accepted it . By contrast to numerous other commensals, C . albicans has a prominent ability to invade virtually all tissues of a host presenting with natural or acquired defects in homeostasis . C . albicans uses considerable energy to synthesize glycans, which are present either as polymers or as glyconjugates . These glycan molecules play a prominent role in the biology of C . albicans by controlling the structure and plasticity of the cell wall, and are also involved in yeast-host interactions . These glycans are recognized as 'non-self' by host innate and adaptative immunity . The signal they induce in the host depends on the 'glycan code', which is determined by the nature of the sugar, the anomer type of linkage and branching, and the length of the oligosaccharide chains . However, this model is not static because the nature of the C . albicans molecule carrying such glycan codes and their expression at the cell wall surface also determines the host response, and, in turn, the regulation of cell wall glycan arrangement dynamics in C . albicans depends on host stimuli . Candida glycans therefore play an important role in the continuous interchange that regulates the balance between saprophytism and parasitism, and resistance and infection . A goal of current research concerning the virulence attributes of C . albicans will be to determine to what extent this species is able to regulate its glycan code as a response to the host.

Curr Opin Microbiol, 2004 Aug, 7(4), 330 - 5
Proteomics on its way to study host-pathogen interaction in Candida albicans; Rupp S; Candida albicans is a common fungal pathogen of humans, but also exists as a commensal in the population . Proteomics of C . albicans has been used since the early 1980s, however, only the recent publication of the genome sequence of C . albicans and improvements in mass spectrometry technologies have made it possible to apply proteomics to C . albicans on a larger scale . This includes analysing the cell wall, investigating drug response or changes in mutants with defects in virulence . In addition, serological responses to systemic candidiasis have been monitored and screens for virulence factors using patient sera, have been described . These promising approaches are just emerging, anticipating further contributions in C . albicans proteomics that will advance our understanding of host-pathogen interaction in the near future.

Front Biosci, 2004 Sep 01, 9, 3248 - 56
A role for Pin1 in mammalian germ cell development and spermatogenesis; Atchison FW et al.; The peptidyl-prolyl isomerase Pin1 is proposed to have diverse functions in many vital aspects of the cell . Despite the multitude of proteins targeted by Pin1 and the proposed regulatory role it plays in critical cellular functions, Pin1 is an essential gene in some eukaryotic organisms, but is dispensable in metazoans . In two genetic models, Candida albicans and Drosophila melanogaster, Pin1 participates in distinct developmental processes regulated by the MAPK pathway . Pin1-deficient mice exhibit decreased primordial germ cell proliferation during embryonic development, along with several degenerative or proliferative defects in the adult testis, retina, mammary gland, and brain . The combination of primordial germ cell deficit and spermatogonial depletion contributes to severe fertility defects in Pin1-null mice . Since growth factor activated MAPK pathways are vital to germ cell proliferation and differentiation, a role for Pin1 in mammalian germ cell development and spermatogenesis is discussed in the context of the Ras/MEK/MAPK pathway.

Bioorg Med Chem, 2004 Oct 1, 12(19), 5107 - 13
Synthesis, antimicrobial, and anti-HIV-1 activity of certain 5-(1-adamantyl)-2-substituted thio-1,3,4-oxadiazoles and 5-(1-adamantyl)-3-substituted aminomethyl-1,3,4-oxadiazoline-2-thiones; El-Emam AA et al.; The reaction of 5-(1-adamantyl)-1,3,4-oxadiazoline-2-thione 2 with iodoethane, 2-dimethylaminoethyl chloride hydrochloride or 2-piperidinoethyl chloride hydrochloride in ethanolic potassium hydroxide yielded the corresponding 5-(1-adamantyl)-2-ethyl or substituted ethylthio-1,3,4-oxadiazoles 3a-c . Interaction of 2 with formaldehyde solution and primary aromatic amines or 1-substituted piperazines, in ethanol at room temperature yielded the corresponding 5-(1-adamantyl)-3-arylaminomethyl-1,3,4-oxadiazoline-2-thiones 4a-m or 5-(1-adamantyl)-3-(4-substituted-1-piperazinylmethyl)-1,3,4-oxadiazoline-2-thiones 5a-h, respectively . All the synthesized compounds were tested for in vitro activities against certain strains of Gram-positive and Gram-negative bacteria and the yeast-like pathogenic fungus Candida albicans . Compounds 2, 5a, and 5e were found as the most active derivatives, particularly against the Gram-positive bacteria . In addition, the antiviral activity of compounds 2, 4a-m, and 5a-h against HIV-1 using the XTT assay was carried out . Compound 2 produced 100%, 43%, and 37% reduction of viral replication at 50, 10, and 2microg/mL concentrations, respectively.

Diagn Cytopathol, 2004 Sep, 31(3), 141 - 6
Rapid review of liquid-based smears as a quality control measure; Henderson S et al.; The objective of this study was to investigate the effectiveness of a standardized method of rapid review (RR) of monolayer preparations for the identification of abnormalities, the presence of an endocervical component and infectious agents . A total of 200 ThinPrep (Cytyc, Boxborough, MA) slides representing the spectrum of abnormalities commonly encountered in cervical/vaginal cytologic specimens was retrieved from archive . The study set comprised 129 cases within normal limits (WNL); 36 low-grade epithelial abnormalities (LGEA); 28 high-grade epithelial abnormalities (HGEA), including 2 endocervical adenocarcinomas in situ (AIS) and 7 carcinomas . Eighteen false negative (FN) cases were also included for study . Originally missed on initial review, these cases were found to be abnormal on quality control review (17 LGEA; 1 AIS) . Commonly encountered infectious agents were represented and included Candida albicans, Trichomonas vaginalis, herpes simplex virus, and Actinomyces . The slides were reviewed using a standardized method of RR (turret technique, for 60 sec) by three experienced screeners masked to the original reference diagnosis . Median sensitivity for LGEA was 70% (range, 67-72%); HGEA, 69% (range, 54-80%); and FN, 65% (range, 56-78%) . Specificity remained high, median specificity for LGEA was 95%; HGEA, 97%; and FN, 100% . There was no significant overcalling of any diagnostic category . The chi-square test at P < 0.05 showed no significant difference between RR and full manual rescreen of the ThinPrep smears in this study . While no statistical difference was proven, the sensitivity measurements for all categories of abnormality were moderate due to the high proportion of atypical cases included into the study set . Abnormalities on the monolayer preparations frequently displayed fewer, smaller groups of disaggregated cells with rounded cytoplasmic outlines that were difficult to discern on RR . Interobserver variation was noted . Monolayers with a paucity of diagnostic cells and those displaying subtle nuclear atypia were often overlooked.

J Mater Sci Mater Med, 2003 Jun, 14(6), 497 - 502
The effect of resin/crosslinker ratio on the mechanical properties and fungal deterioration of a maxillofacial silicone elastomer; Taylor RL et al.; Variation of the crosslinker/resin ratio of a room temperature condensation cure maxillofacial silicone elastomer has caused considerable changes in the mechanical properties and deterioration by Candida albicans . Increasing the crosslinker/resin ratio caused a decrease in the tensile strength and stiffness of the elastomer . However, tear strength appeared to show an optimum value at the recommended crosslinker/resin ratio . These effects were due to the low molar mass silicone polymer that acts as a carrier for the actual crosslinking additive . The general decrease in mechanical properties was accompanied by an increase in the hexane extractables content and an increase in the Si-H content of the elastomer . The unbound polymer (extractable material) content of the elastomer was found to influence the colonization of the material by C . albicans . An increase in the unbound polymer content corresponded to an increasing number of hyphae and blastospores observed penetrating into the elastomer . The data obtained in this study have significant implications concerning the degree of control of elastomer formulation and the deterioration of maxillofacial appliances.

Microbiology, 2004 Sep, 150(Pt 9), 3041 - 9
Identification of the dialysable serum inducer of germ-tube formation in Candida albicans; Hudson DA et al.; Yeast cells of Candida albicans are induced by serum at 37 degrees C to produce germ tubes, the first step in a transition from yeast to hyphal growth . Previously, it has been shown that the active component is not serum albumin but is present in the dialysable fraction of serum . In this study, serum induction of germ-tube formation is shown to occur even in the presence of added exogenous nitrogen sources and is therefore not signalled by nitrogen derepression . The active component in serum was purified by ion-exchange, reverse-phase and size-exclusion chromatography from the dialysable fraction of serum and was identified by NMR to be d-glucose . Enzymic destruction of glucose, using glucose oxidase, demonstrated that d-glucose was the only active component in these fractions . Induction of germ-tube formation by d-glucose required a temperature of 37 degrees C and the pH optimum was between pH 7.0 and 8.0 . d-Glucose induced germ-tube formation in a panel of clinical isolates of C . albicans . Although d-glucose is the major inducer in serum, a second non-dialysable, trichloroacetic acid precipitable inducer is also present . However, whereas either 1.4 % (v/v) serum or an equivalent concentration of d-glucose induced 50 % germ-tube formation, the non-dialysable component required a 10-fold higher concentration to induce 50 % germ-tube formation . Serum is, therefore, the most effective induction medium for germ-tube formation because it is buffered at about pH 8.5 and contains two distinct inducers (glucose and a non-dialysable component), both active at this pH.

Microbiology, 2004 Sep, 150(Pt 9), 2807 - 13
Polymorphonuclear leukocytes (PMNs) induce protective Th1-type cytokine epithelial responses in an in vitro model of oral candidosis; Schaller M et al.; The immune response and the anticandidal activity of keratinocytes and polymorphonuclear leukocytes (PMNs) play a key role in host defence against localized Candida albicans infection . An established model of oral candidosis based on reconstituted human oral epithelium (RHE) was supplemented with PMNs to study the effect of these immune cells during experimental oral candidosis . Infection of RHE with C . albicans induced a strong expression of the chemokine interleukin-8 (IL-8) and the cytokine granulocyte-macrophages colony-stimulating factor (GM-CSF), and a moderate stimulation of interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interferon gamma (IFN-gamma) and tumour necrosis factor alpha (TNF-alpha) by keratinocytes . This immune response was associated with chemoattraction of PMNs to the site of infection, whereas uninfected RHE failed to induce cytokine expression or to attract PMNs . Growth of the pathogen and tissue damage of C . albicans-infected RHE were significantly reduced when PMNs were applied to the apical epithelial surface or when PMNs migrated through a perforated basal polycarbonate filter of the model . Notably, protection against epithelial tissue damage was also observed when PMNs were placed on the basal side of non-perforated filters, which prevented PMN migration into the RHE . Addition of PMNs enhanced a Th1-type immune response (IFN-gamma, TNF-alpha), down-regulated the expression of the Th2-type cytokine interleukin-10 (IL-10), and was associated with protection against Candida-induced tissue damage . This PMN-supplemented model of oral candidosis mimics the in vivo situation, and provides a promising tool for studying the immunological interactions between keratinocytes and C . albicans, as well as the influence of PMNs on C . albicans pathogenesis.

J Biol Chem, 2004 Nov 12, 279(46), 47952 - 60 Epub 2004 Sep 03.
Inactivation of CaMIT1 inhibits Candida albicans phospholipomannan beta-mannosylation, reduces virulence, and alters cell wall protein beta-mannosylation; Mille C et al.; Studies on Candida albicans phospholipomannan have suggested a novel biosynthetic pathway for yeast glycosphingolipids . This pathway is thought to diverge from the usual pathway at the mannose-inositol-phospho-ceramide (MIPC) step . To confirm this hypothesis, a C . albicans gene homologue for the Saccharomyces cerevisiae SUR1 gene was identified and named MIT1 as it coded for GDP-mannose:inositol-phospho-ceramide mannose transferase . Two copies of this gene were disrupted . Western blots of cell extracts revealed that strain mit1Delta contained no PLM . Thin layer chromatography and mass spectrometry confirmed that mit1Delta did not synthesize MIPC, demonstrating a role of MIT1 in the mannosylation of C . albicans IPCs . As MIT1 disruption prevented downstream beta-1,2 mannosylation, mit1Delta represents a new C . albicans mutant affected in the expression of these specific virulence attributes, which act as adhesins/immunomodulators . mit1Delta was less virulent during both the acute and chronic phases of systemic infection in mice (75 and 50% reduction in mortality, respectively) . In vitro, mit1Delta was not able to escape macrophage lysis through down-regulation of the ERK1/2 phosphorylation pathway previously shown to be triggered by PLM . Phenotypic analysis also revealed pleiotropic effects of MIT1 disruption . The most striking observation was a reduced beta-mannosylation of phosphopeptidomannan . Increased beta-mannosylation of mannoproteins was observed under growth conditions that prevented the association of beta-oligomannosides with phosphopeptidomannan, but not with PLM . This suggests that C . albicans has strong regulatory mechanisms associating beta-oligomannoses with different cell wall carrier molecules . These mechanisms and the impact of the different presentations of beta-oligomannoses on the host response need to be defined.

Shi Yan Sheng Wu Xue Bao, 2002 Dec, 35(4), 289 - 95
{Long terminal repeats of retroposons in Candida albicans}; Wang Q et al.; 25 Candida albicans strains have been analysed on Southern hybridization patterns with alpha, beta, gamma, kappa probes . The alpha, beta, gamma, kappa, elements can be used to distinguish the differences and relations between C . albicans strains . Chromosomal localization analysis indicated that the alpha, beta, gamma, kappa, elements were located on several different chromosomes in Candida albicans strains . Northern hybridization showed that alpha, beta, gamma were transcribed actively and regulated by different stimuli . The kappa was inactive . Our results indicated that in Candida albicans there were many kinds of retroposons whose numbers and distribution were different . These retroposons could be used to identify Candida albicans strains as moleculer markers.

J Infect Dis, 2004 Oct 1, 190(7), 1327 - 34 Epub 2004 Aug 20.
beta-defensin expression in immunocompetent and immunodeficient germ-free and Candida albicans-monoassociated mice; Schofield DA et al.; BACKGROUND: Defensins are important components of innate immunity and play a key role in the fight against infectious diseases; however, little is known about their role in resistance to fungal infection . METHODS: We examined gene expression of mouse beta -defensins (mBDs) 1-4 in orogastric tissues from germ-free (gf) and Candida albicans-monoassociated immunocompetent (C57BL/6) and immunodeficient (Tg varepsilon 26 or gp91(phox-/-)/NOS2(-/-)) mice, using competitive reverse-transcriptase polymerase chain reaction . RESULTS: The basal levels of beta -defensin gene expression in orogastric tissues from gf mice varied significantly between immunodeficient and immunocompetent mice and by the particular tissue analyzed . During gastric and lethal oroesophageal candidiasis, expression of mBD1, -3, and -4 was induced at the infection sites (stomach and tongue) and was concomitant with an induction of tumor necrosis factor- alpha expression in Tg varepsilon 26 mice, compared with that in tissues from gf mice . Induction of mBD4 expression in response to gastric candidiasis, however, was dependent on the immune competency of the host . A C . albicans mutant that lacked key genes important for filamentation and virulence also significantly induced expression of mBD1, -3, and -4 in Tg varepsilon 26 mice . CONCLUSIONS: These data not only demonstrate quantitative and qualitative differences in beta -defensin expression in gf and gnotobiotic mice, they also suggest a role for these peptides in resistance to gastric and lethal oroesophageal candidiasis.

J Infect Dis, 2004 Oct 1, 190(7), 1318 - 26 Epub 2004 Aug 25.
Induction of nuclear factor- kappa B and c-Jun/activator protein-1 via toll-like receptor 2 in macrophages by antimycotic-treated Candida albicans; Roeder A et al.; We examined the role of Toll-like receptors (TLRs) by using TLR2-deficient (TLR2(-/-)), TLR4-defective (TLR4(d/d)), and double-knockout murine macrophages and human embryonic kidney (HEK) 293 cells transfected with human TLR2 or TLR4 expression plasmids after stimulation with different preparations of the human pathogenic fungus Candida albicans . Compared with wild-type macrophages, TLR2(-/-) and TLR4(d/d) macrophages had impaired recognition of viable C . albicans, whereas antimycotic (AM)-treated C . albicans solely used TLR2 in a TLR4- and interferon- gamma -independent manner . In human HEK293 cells, AM-treated C . albicans elicited mainly TLR2-dependent activation . The differences in responsiveness to viable C . albicans, compared with C . albicans treated with cytoplasmic membrane-interacting AMs, suggest specific recognition of different pathogen-associated patterns by TLRs in innate antifungal responses . Our analyses of signal transduction after stimulation of wild-type macrophages with AM-treated C . albicans demonstrated involvement of the transcription factors nuclear factor- kappa B and c-Jun/activator protein-1 and of the mitogen-activated protein kinases p38, extracellular-related kinase, and c-Jun NH(2)-terminal kinase.

Microbes Infect, 2004 Sep, 6(11), 985 - 9
Human dendritic cells are less potent at killing Candida albicans than both monocytes and macrophages; Netea MG et al.; Dendritic cells (DC) function as professional phagocytes to kill Candida albicans and subsequently present it to the adaptive immune system . Monocytes, macrophages and DC were generated from five individual donors and their Candida-killing capacity and cytokine release were assessed . Compared to monocytes and macrophages, DC from healthy volunteers were significantly less effective in C . albicans--stimulated cytokine release, killing of C . albicans blastoconidia and damaging of C . albicans hyphae . In conclusion, while important as antigen-presenting cells and initiators of the adaptive immune system, DC are poor in both intracellular killing and damaging of C . albicans hyphae . Effective handling of large numbers of C . albicans is the prime task of the innate immune system consisting of large numbers of neutrophils and monocytes.

Fungal Genet Biol, 2004 Oct, 41(10), 941 - 53
Analysis of heterologous expression of Candida albicans SEC61 gene reveals differences in Sec61p homologues related to species-specific functionality; de la Rosa JM et al.; The protein secretory pathway has not been studied in depth in Candida albicans despite its essential role in the secretion of enzymes and cell surface components related to the ability of the fungus to colonize the human host . To gain further insight into the elements that participate in the first stages of the secretory process in this fungal pathogen we have isolated and characterized the C . albicans ortholog of SEC61 . In other species SEC61 has been shown to encode the core element of the protein translocation apparatus within the ER membrane . The cloned gene appears to be essential for cell viability and encodes a highly conserved protein, very similar to the Sec61p from other yeast species both in sequence and hydropathy profile . However, CaSec61p is not able to complement the thermosensitive-growth phenotype of a Saccharomyces cerevisiae sec61 mutant, even though it is expressed and correctly incorporated into the ER membrane of the transformant cells . We report results indicating that the lack of functional complementation could be related to differences in the primary structure of the cytosolic domain located between the fourth and fifth transmembrane domains of the accepted topological model of Sec61p.

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 2004 Jun, 18(2), 179 - 80
{A prospective study on short period antibiotic treatment of hepatic failure complicated with spontaneous bacterial peritonitis}; Li HW et al.; BACKGROUND: To observe the effects of short-term antibiotic treatment in patients with hepatic failure and spontaneous bacterial peritonitis (SBP) . METHODS: In this prospective study short-term antibiotic treatment was given to 67 cases diagnosed as hepatic failure with spontaneous bacterial peritonitis . Ceftriaxone 2 g, iv drip, q12h for 10 d or ofloxacin 0.2 g, iv drip, q12h for 10 d was given to the patients at random and the efficacy was evaluated on the basis of clinical symptoms, medical examination and ascites after 3, 7, 10 days of therapy . RESULTS: Seven cases (10.44%) were cured and 57 cases (85%) were improved after 3 days therapy, the total effective rate was 95.52%, but in 3 cases the therapy had no effect . The results of ascites bacterial culture and drug susceptibility test showed that one case had drug resistance to ceftriaxone and two cases had drug resistance to ofloxacin, so antibiotics were changed in time . After 7 days therapy, the results showed that 65 cases (97.01%) cured and 2 cases (2.99%) were improved, the total effective rate was 100% . When the therapy lasted for 10 days, all patients were cured . One patient had oral mucous membrane . Candida albicans infection after 3 days therapy; two cases got thrush and one patient got fungal intestinal infection after 7 days therapy; when the therapy lasted for 10 days, 4 cases had thrush and 2 cases had fungal infection of intestines and one patient had pulmonary fungal infection . CONCLUSION: The optimal period of antibiotic treatment of hepatic failure with SBP should be from 7 days to 10 days.

Mol Cell Biol, 2004 Sep, 24(18), 7949 - 57
Innate immune responses in peptidoglycan recognition protein L-deficient mice; Xu M et al.; Peptidoglycan recognition proteins (PGRPs) constitute a family of innate immune recognition molecules . In Drosophila, distinct PGRPs bind to peptidoglycans on gram-positive or gram-negative bacteria and provide essential signals upstream of the Toll and Imd pathways required for immunity against infection . Four PGRPs, PGRP-L, -S, -Ialpha, and -Ibeta, are expressed from three genes in mammals . In this paper, we provide direct evidence that the longest family member, PGRP-L, is a secreted serum protein with the capacity to multimerize . Using gene targeting to create PGRP-L-deficient mice, we demonstrate little contribution by PGRP-L to systemic challenge using gram-negative bacteria (Escherichia coli, slightly less susceptible), Gram-positive bacteria (Staphylococcus aureus), or yeast (Candida albicans) . Peritoneal macrophages from PGRP-L-deficient mice produced decreased amounts of the inflammatory cytokines interleukin 6 and tumor necrosis factor alpha when stimulated with E . coli or lipopolysaccharide, but comparable amounts when stimulated with S . aureus, C . albicans, or their cell wall components . Additionally, these cells produced similar amounts of cytokines when challenged with gram-positive or -negative peptidoglycans . In contrast to its critical role in immunity in flies, PGRP-L is largely dispensable for mammalian immunity against bacteria and fungi.

Cell Microbiol, 2004 Oct, 6(10), 915 - 26
Candida albicans proteinases and host/pathogen interactions; Naglik J et al.; Candida infections are common, debilitating and often recurring fungal diseases and a problem of significant clinical importance . Candida albicans, the most virulent of the Candida spp., can cause severe mucosal and life-threatening systemic infections in immunocompromised hosts . Attributes that contribute to C . albicans virulence include adhesion, hyphal formation, phenotypic switching and extracellular hydrolytic enzyme production . The extracellular hydrolytic enzymes, especially the secreted aspartyl proteinases (Saps), are one of few gene products that have been shown to directly contribute to C . albicans pathogenicity . Because C . albicans is able to colonize and infect almost every tissue in the human host, it may be crucial for the fungus to possess a number of similar but independently regulated and functionally distinct secreted proteinases to provide sufficient flexibility in order to survive and promote infection at different niche sites . The aim of this review is to explore the functional roles of the C . albicans proteinases and how they may contribute to the host/pathogen interaction in vivo.

East Mediterr Health J, 2001 Nov, 7(6), 925 - 34
Gram stain versus culture in the diagnosis of vulvovaginal candidiasis; Omar AA; We evaluated Gram stain as a rapid diagnostic method for vulvovaginal candidiasis (VVC) . Vaginal swabs were taken from 100 pregnant women and subjected to Gram stain and culture; 39% were diagnosed as having VVC (29% had symptomatic VVC and 10% had asymptomatic VVC) . Candida albicans was isolated from 94.9% of all cultures while other Candida species were isolated from 5.1% of cases . Multigravidae were significantly more affected than primigravidae . Gram stain is a valuable method in rapid accurate diagnosis of symptomatic VVC and even superior to culture as it demonstrates the invasive forms of the yeast . However, its low sensitivity could restrict its use in routine practice . A combination of culture and Gram stain is the ideal approach for the diagnosis of VVC.

J Chemother, 2004 Jun, 16(3), 255 - 8
Combined effect of liposomal and conventional amphotericin B in a mouse model of systemic infection with Candida albicans; Kretschmar M et al.; Amphotericin B may be employed as a potentially toxic deoxycholate complex or incorporated in liposomes and other particles which have an altered tissue distribution . A combination of both preparations could help to overcome the drawbacks of the individual preparations . We have employed a mouse model of systemic infection with Candida albicans to investigate whether this assumption was essentially true . We found that the combination of low dosages of both preparations (0.5 mg/kg of body weight/day of conventional and 0.5 mg kg of body weight/day liposomal amphotericin B) was more efficient than a similar dosage of conventional amphotericin B (1 mg/kg of body weight/day) in the liver and similar or higher dosages of liposomal amphotericin B (1 or 5 mg/kg of body weight/day) in the kidneys.

J Mater Sci Mater Med, 2004 Feb, 15(2), 117 - 21
Glass fibre-reinforced composite laced with chlorhexidine digluconate and yeast adhesion; Waltimo T et al.; The aim of this study was to lace dental glass fibre reinforced composite (FRC) prepreg with chlorhexidine digluconate and to examine the adherence of common oral fungal pathogen Candida albicans to FRC made of the prepreg . Four different test and control material groups each comprising 16 test specimens ((5.0 x 5.0 x 0.8) mm3) each were used as substrates for C . albicans adherence . A porous polymer pre-impregnated woven glass fibre prepreg was laced with solution of chlorhexidine gluconate and it was used with autopolymerized denture base polymer to fabricate FRC test specimens . Control group (Group 1) consisted of FRC test specimens stored in water . In Group 2, the test specimens were stored in 10% chlorhexidine digluconate solution for 24 h . Group 3 consisted of specimens fabricated using such fibre reinforcements which were pre-soaked in 20% chlorhexidine digluconate and dried before preparation with denture base resin, and followed by storage of the specimens in water . Group 4 was similar to Group 3 but instead of water storage the specimens were immersed in 10% chlorhexidine digluconate for 24 h . For the candidal adhesion assay the test and control specimens were incubated in standardized suspensions of four different strains of C . albicans, rinsed and prepared for light-microscopy . The mean number of adherent cells in each group was counted microscopically and analysed statistically . There were significantly (P < 0.05) more adherent C . albicans cells found in Group 1 than in the other three groups which did not differ significantly from each other . The lowest numbers of adherent cells were found in Group 3 . Pretreating the porous polymer pre-impregnated glass fibre reinforcement with chlorhexidine digluconate result in reduction in the number of adherent yeast cells on the surface FRC material.

N Engl J Med, 2004 Aug 26, 351(9), 876 - 83
Maintenance fluconazole therapy for recurrent vulvovaginal candidiasis; Sobel JD et al.; BACKGROUND: No safe and convenient regimen has proved to be effective for the management of recurrent vulvovaginal candidiasis . METHODS: After inducing clinical remission with open-label fluconazole given in three 150-mg doses at 72-hour intervals, we randomly assigned 387 women with recurrent vulvovaginal candidiasis to receive treatment with fluconazole (150 mg) or placebo weekly for six months, followed by six months of observation without therapy . The primary outcome measure was the proportion of women in clinical remission at the end of the first six-month period . Secondary efficacy measures were the clinical outcome at 12 months, vaginal mycologic status, and time to recurrence on the basis of Kaplan-Meier analysis . RESULTS: Weekly treatment with fluconazole was effective in preventing symptomatic vulvovaginal candidiasis . The proportions of women who remained disease-free at 6, 9, and 12 months in the fluconazole group were 90.8 percent, 73.2 percent, and 42.9 percent, as compared with 35.9 percent, 27.8 percent, and 21.9 percent, respectively, in the placebo group (P< 0.001) . The median time to clinical recurrence in the fluconazole group was 10.2 months, as compared with 4.0 months in the placebo group (P<0.001) . There was no evidence of fluconazole resistance in isolates of Candida albicans or of superinfection with C . glabrata . Fluconazole was discontinued in one patient because of headache . CONCLUSIONS: Long-term weekly treatment with fluconazole can reduce the rate of recurrence of symptomatic vulvovaginal candidiasis . However, a long-term cure remains difficult to achieve .

J Mol Biol, 2004 Aug 20, 341(4), 1085 - 96
Potential anti-infective targets in pathogenic yeasts: structure and properties of 3,4-dihydroxy-2-butanone 4-phosphate synthase of Candida albicans; Echt S et al.; A synthetic gene specifying a putative 3,4-dihydroxy-2-butanone 4-phosphate synthase of Candida albicans directed the synthesis of a 22.5 kDa peptide in a recombinant Escherichia coli strain . The recombinant protein was purified to apparent homogeneity by two chromatographic steps and was shown to catalyze the formation of L-3,4-dihydroxy-2-butanone 4-phosphate from ribulose 5-phosphate at a rate of 332 nmol mg(-1) min(-1) . Hydrodynamic studies indicated a native molecular mass of 41 kDa in line with a homodimer structure . The protein was crystallized in its apoform . Soaking yielded crystals in complex with the substrate ribulose 5-phosphate . The structures were solved at resolutions of 1.6 and 1.7 angstroms, respectively, using 3,4-dihydroxy-2-butanone 4-phosphate synthase of E . coli for molecular replacement . Structural comparison with the orthologs of Magnaporthe grisea and Methanococcus jannaschii revealed a hitherto unknown conformation of the essential acidic active-site loop .

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3425 - 35
Disruption of the Candida albicans CYB5 gene results in increased azole sensitivity; Rogers KM et al.; Sterol synthesis in fungi is an aerobic process requiring molecular oxygen and, for several cytochrome-mediated reactions, aerobically synthesized heme . Cytochrome b(5) is required for sterol C5-6 desaturation and the encoding gene, CYB5, is nonessential in Saccharomyces cerevisiae . Cyb5p and Ncp1p (cytochrome P-450 reductase) appear to have overlapping functions in this organism, with disruptions of each alone being viable . The cytochrome P-450 reductase phenotype has also been shown to demonstrate increased sensitivity to azole antifungals . Based on this phenotype, the CYB5 gene in the human pathogen Candida albicans was investigated to determine whether the cyb5 genotype was viable and would also demonstrate azole sensitivity . Sequential disruption of the CYB5 alleles by direct transformation resulted in viability, presumably conferred by the presence of a third copy of the CYB5 gene . Subsequent disruption procedures with a pMAL2-CYB5 rescue cassette and a CYB5-URA3 blaster cassette resulted in viable cyb5 strains with no third copy . The C . albicans CYB5 gene is concluded to be nonessential . Thus, the essentiality of this gene and whether we observed two or three alleles was dependent upon the gene disruption protocol . The C . albicans cyb5 strains produced a sterol profile containing low ergosterol levels and sterol intermediates similar to that reported for the S . cerevisiae cyb5 . The C . albicans cyb5 shows increased sensitivity to azoles and terbinafine, an inhibitor of squalene epoxidase, and, unexpectedly, increased resistance to morpholines, which inhibit the ERG2 and ERG24 gene products . These results indicate that an inhibitor of Cyb5p would not be lethal but would make the cell significantly more sensitive to azole treatment.

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