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-P