Infect Immun, 2004 Apr, 72(4), 2029 - 34 Degenerate peptide recognition by Candida albicans adhesins Als5p and Als1p; Klotz SA et al.; Candida albicans and Saccharomyces cerevisiae expressing the adhesins Als5p or Als1p adhere to immobilized peptides and proteins that possess appropriate sequences of amino acids in addition to a sterically accessible peptide backbone . In an attempt to further define the nature of these targets, we surveyed the ability of yeast cells to adhere to 90- micro m-diameter polyethylene glycol beads coated with a 7-mer peptide from a library of 19(7) unique peptide-beads . C . albicans bound to ca . 10% of beads from the library, whereas S . cerevisiae expressing Als5p or Als1p bound to ca . 0.1 to 1% of randomly selected peptide-beads . S . cerevisiae expressing Als1p had a distinctly different adherence phenotype than did cells expressing Als5p . The former adhered in groups or clumps of cells, whereas the latter adhered initially as single cells, an event which was followed by the build up of cell-cell aggregates . Beads with adherent cells were removed, and the peptide attached to the bead was determined by amino acid sequencing . All adhesive beads carried a three-amino-acid sequence motif (tau phi+) that possessed a vast combinatorial potential . Adherence was sequence specific and was inhibited when soluble peptide identical to the immobilized peptide was added . The Als5p adhesin recognized some peptides that went unrecognized by Als1p . The sequence motif of adhesive peptides identified by this method is common in proteins and offers so many possible sequence combinations that target recognition by the Als proteins is clearly degenerate . A degenerate recognition system provides the fungi with the potential of adhering to a multitude of proteins and peptides, an advantage for any microorganism attempting to establish a commensal or pathogenic relationship with a host. Infect Immun, 2004 Apr, 72(4), 1956 - 63 Salivary secretory leukocyte protease inhibitor and oral candidiasis in human immunodeficiency virus type 1-infected persons; Chattopadhyay A et al.; Oropharyngeal candidiasis, typically caused by Candida albicans, is the most common oral disease associated with human immunodeficiency virus type 1 (HIV-1) infection . Secretory leukocyte protease inhibitor (SLPI), a 12-kDa antiprotease, suppresses the growth of C . albicans in vitro . To determine whether the mucosal protein plays a role in protecting oral tissues against fungal infection, we conducted a cross-sectional study investigating the oral and systemic health and salivary SLPI levels in 91 dentate HIV-1-infected adults receiving medical care in the southeastern United States . Participants with a self-reported history of clinical oropharyngeal candidiasis during the previous 2 years constituted the test group (n = 52), while the comparison group (n = 39) had no oropharyngeal candidiasis during that period . Data collected from medical records, oral examination, and SLPI enzyme-linked immunosorbent assay quantitation of whole saliva were analyzed by t test, analysis of variance, linear regression, and unconditional logistic regression . The test group had a significantly higher mean salivary SLPI level than the comparison group (1.9 microg/ml versus 1.1 microg/ml, P < 0.05) . Linear regression modeling identified CD4 cell count and history of oropharyngeal candidiasis as key predictors of salivary SLPI and revealed a significant interaction (P < 0.05) between immunosuppression (CD4 cell count below 200 cells/ microl) and positive history of oropharyngeal candidiasis in predicting salivary SLPI level . By logistic regression modeling, a salivary SLPI level exceeding 2.1 microg/ml, low CD4 count, antiretroviral monotherapy, and smoking were key predictors of oropharyngeal candidiasis . These data support a key role for SLPI in the oral mucosal defense against C . albicans . The antimicrobial mucosal protein may serve as an indicator of previous oropharyngeal candidiasis infection among immunosuppressed persons. FEMS Immunol Med Microbiol, 2004 Apr 9, 40(3), 239 - 42 LAAE-14, a new anti-inflammatory drug, increases the survival of Candida albicans-inoculated mice; Lucas R et al.; LAAE-14, a lipidic acid-amido ether derivative, has been recently described as a new anti-inflammatory drug . We have studied the effect of treatment with this compound on the susceptibility of mice to in vivo experimental Candida albicans infection . ICR mice orally treated with LAAE-14 (25 mg kg(-1)) and experimentally intravenously infected showed a significantly increased survival as compared to control mice . In vitro, the compound did not inhibit the growth of C . albicans yeast cells or the yeast-to-hyphal transition . The in vitro production of prostaglandin E2 by peritoneal macrophages in response to the yeasts and hyphae of C . albicans was significantly decreased upon treatment with LAAE-14, in a dose-dependent manner . Thus, reduced prostaglandin production during fungal infection could be an important factor in controlling fungal colonisation and infection. Int Immunopharmacol, 2004 Mar, 4(3), 387 - 401 DNA array analysis of altered gene expression in human leukocytes stimulated with soluble and particulate forms of Candida cell wall beta-glucan; Ishibashi K et al.; We previously reported that 1,3-beta-d-glucan derived from Candida albicans, a pathogenic fungus was obtained by oxidation of the cell wall with sodium hypochlorite (NaClO) . It could be solubilized by treatment with dimethylsulfoxide (DMSO) . We found that OX-CA and CSBG showed significantly different levels of activity toward leukocytes . Here, we have used cDNA microarrays to analyze the mRNA expression of 1176 genes in PBMCs stimulated with Candida cell wall glucan and considered the difference in the activation mechanism of OX-CA and CSBG . Total mRNA showed a significant change for 147 out of 1176 arrayed genes on stimulation with OX-CA and CSBG for 4 h . Among those genes, 62 were common, 26 were OX-CA-specific and 59 were CSBG-specific . Many of these up-regulated genes encode effectors with well-characterized proinflammatory properties . The expression of genes related with signal transduction differed in the particulate and soluble glucans derived from C . albicans having exactly the same primary structure . This fact suggested that each glucan induced specific biological activity through a different activation mechanism . This study using cDNA microarrays to analyze a broad spectrum of mRNA expression provides information on the biological activity of Candida cell wall glucan as a potential pathogenic factor. Nat Rev Microbiol, 2003 Nov, 1(2), 106 - 16 The biology of mating in Candida albicans; Johnson A; Candida albicans has maintained an elaborate--but largely hidden--mating apparatus, which shares some features with the closely related 'model' yeast Saccharomyces cerevisiae, but which also has some important differences . The differences are particularly noteworthy, as they could indicate the strategies that allow C . albicans to survive and mate in the hostile environment of a mammalian host . Indeed, some features of C . albicans mating seem to be intimately connected to its host. Curr Genet, 2004 Jun, 45(6), 390 - 8 Epub 2004 Mar 18. Shuttle vectors for Candida albicans: control of plasmid copy number and elevated expression of cloned genes; Du W et al.; Plasmids containing the inosine monophosphate dehydrogenase gene CaIMH3 from Candida albicans strain ATCC 32354 transform their host to resistance against mycophenolic acid (MPA) . The transformants maintain the plasmids at a high copy number (20-40 per cell) and express the CaIMH3 gene at very high levels relative to untransformed controls . The plasmid copy number can be controlled by the concentration of MPA in the media . The transformation procedure is reproducible and the efficiency of transformation is high, up to 15,000 per microgram . Unrearranged plasmids are readily recovered by transforming total DNA from transformants back into Escherichia coli . C . albicans genes cloned into the plasmid are expressed at elevated levels relative to untransformed controls . A derivative vector containing the CaMAL2 promoter and termination sequences expresses the CaERG11 ORF at high levels and confers moderate resistance to fluconazole . These shuttle vectors should facilitate global genomics approaches in C . albicans that have been hampered by its diploid genome. Ann N Y Acad Sci, 2003 Dec, 1010, 573 - 6 Role of phospholipomannan in Candida albicans escape from macrophages and induction of cell apoptosis through regulation of bad phosphorylation; Ibata-Ombetta S et al.; Candida albicans, the most common opportunistic fungal pathogen of humans is a part of the normal microbial flora . To investigate host-parasite interaction related to the commensal-pathogen switch of this yeast we compared the response of macrophages to C . albicans and to the non-pathogenic yeast Saccharomyces cerevisiae . In contrast to S . cerevisiae, C . albicans survived within macrophages . This escape from macrophages was associated with qualitative differences in the sequential phosphorylation of MEK, ERK1/2, and p90RSK during phagocytosis . Decreased activation of this pathway was observed with C . albicans and was associated with a species-specific overexpression of the MEK phosphatase, MKP-1 . Dysregulation of the ERK1/2/p90RSK signal transduction pathway by C . albicans was associated downstream with reduction in Bad phosphorylation, specifically at Ser-112, and disappearance of free Bcl-2 . This ended at apoptosis of cells that have ingested C . albicans, as revealed by staining of phosphatidylserine exposure in the macrophage outer membrane . The role of phospholipomannan (PLM), a phylogenetically unique glycolipid with a phytoceramide moiety expressed at the surface of and shed by C . albicans, was examined . Addition of PLM to macrophages led to dysregulation similar to that observed with live C . albicans and promoted the survival of the sensitive S . cerevisiae within the cells . Evidence of externalization of membranous phosphatidylserine, loss of mitochondrial integrity, and DNA fragmentation after incubation of macrophages with PLM suggest that this molecule supported the activities observed with C . albicans yeast cells. FEMS Microbiol Lett, 2004 Mar 19, 232(2), 133 - 8 The C-terminal antibody binding domain of Candida albicans mp58 represents a protective epitope during candidiasis; Viudes A et al.; The 58-kDa surface mannoprotein of Candida albicans (mp58) elicits strong antibody responses during infection . Epitope mapping with sera from patients with candidiasis and control individuals indicated the presence of multiple IgG-reactive continuous epitopes on the protein, expanding both the amino- and carboxy-terminal domains and several internal regions . These immunoreactive regions were similar to the ones previously identified using sera from immunized animals . Two of the epitopic regions (including the C-terminal domain) showed increased reactivity with antibodies present in sera from patients with candidiasis as compared to control individuals . Patients who survived the infection displayed increased antibody reactivity towards the C-terminal epitope as compared to those succumbing to candidiasis . A monoclonal antibody directed towards this epitopic region conferred protection in serum therapy experiments in a murine model of hematogenously disseminated candidiasis . Together, these observations indicate the carboxy-terminal antibody binding domain of C . albicans mp58 represents a protective epitope during candidiasis. Asian Pac J Allergy Immunol, 2003 Sep, 21(3), 189 - 92 Anergy testing in patients with head and neck cancer; Pumhirun P et al.; Total T cells, but the reasons why the patients became anergic . Patients with head and neck cancer were found to be deficient in were not clear . Possible explanations include a change in T-lymphocyte numbers, particularly the helper/suppressor T-cell ratio, with the cause of this change still unknown . Tumor immunosuppressing factors and cancer-induced immunosuppression are proposed to be such causes . The deficiency of T cells resulted in an impaired cell-mediated immune response (CMIR), which lowered the host resistance, such facilitating the tumor to spread . As the CMIR can be evaluated by delayed hypersensitivity skin testing (= anergy screen), the objective of this study was to compare the CMIR function of patients with head and neck cancer to a non-cancer control group using this anergy screen . The study group consisted of 20 patients (17 males, 3 females, age range 10-76 years) with head and neck cancer, which were anti-HIV negative and had not received any therapy yet . The control group consisted of another 20 persons (17 males, 3 females, age range 21-72 years) without any cancer and who were also anti-HIV negative . Exclusion criteria were (1) eczema or skin disease in the area to be tested, (2) having received oral prednisolone within the last week and (3) an anti-HIV positive immune status . The antigens used in this study consisted of PPD (5 IU), tetanus toxoid (TT) (0.8 LF/ml and 1.6 LF/ml, Candida albicans (20 PNU/ ml and 200 PNU/ml), mumps-measles-rubella (MMR) vaccine (1:10 v/v and 1:5 v/v) . The test was done by intradermal injection of 0.1 ml of each antigen . The anergy screen was considered positive when the test resulted in an erythema or induration larger than 5 mm at 72 hours after the injection . Complete anergy was diagnosed when there was no skin reaction at all, partial anergy when only 1 antigen tested positive and no anergy when there were positive skin reactions to two or more antigens . In the study group, 9 (45%) patients were diagnosed with complete anergy, 11 (55%) with partial anergy and none with no anergy, while in the control group, none were complete anergic, 3 (15%) were partially anergic and 17 (85%) had no anergy . There was a statistically significant difference (p < 0.01) between these two groups . In conclusion, patients with head and neck cancer seemed to have an impaired CMIR, with at least the partial anergy being statistical significantly different compared to the non-cancer group. Antonie Van Leeuwenhoek, 2004 Apr, 85(3), 199 - 207 Partial purification and characterization of a mannosyl transferase involved in O -linked mannosylation of glycoproteins in Candida albicans; Arroyo-Flores BL et al.; Incubation of a mixed membrane fraction of C . albicans with the nonionic detergents Nonidet P-40 or Lubrol solubilized a fraction that catalyzed the transfer of mannose either from endogenously generated or exogenously added dolichol-P-{14C}Man onto endogenous protein acceptors . The protein mannosyl transferase solubilized with Nonidet P-40 was partially purified by a single step of preparative nondenaturing electrophoresis and some of its properties were investigated . Although transfer activity occurred in the absence of exogenous mannose acceptors and thus depended on acceptor proteins isolated along with the enzyme, addition of the protein fraction obtained after chemical de-mannosylation of glycoproteins synthesized in vitro stimulated mannoprotein labeling in a concentration-dependent manner . Other de-mannosylated glycoproteins, such as yeast invertase or glycoproteins extracted from C . albicans, failed to increase the amount of labeled mannoproteins . Mannosyl transfer activity was not influenced by common metal ions such as Mg(2+), Mn(2+) and Ca(2+), but it was stimulated up to 3-fold by EDTA . Common phosphoglycerides such as phosphatidylglycerol and, to a lower extent, phosphatidylinositol and phosphatidylcholine enhanced transfer activity . Interestingly, coupled transfer activity between dolichol phosphate mannose synthase, i.e., the enzyme responsible for Dol-P-Man synthesis, and protein mannosyl transferase could be reconstituted in vitro from the partially purified transferases, indicating that this process can occur in the absence of cell membranes. Fitoterapia, 2004 Mar, 75(2), 192 - 200 Antimicrobial activity of Artemisia douglasiana leaf essential oil; Setzer WN et al.; Artemisia douglasiana leaf has been shown to be efficacious complementary herbal treatment for chronic bladder infection in a paraplegic youth . The leaf oil has been analyzed by GC-MS and the major components found to be camphor (29%), artemisia ketone (26%), artemisia alcohol (13%), alpha-thujone (10%), 1,8-cineole (8%), and hexanal (5%) . The leaf oil and the major components have been tested for antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger . The essential oil shows limited antimicrobial activity in vitro, so it is unclear if the oil exerts a direct antimicrobial effect in vivo, or plays some role in stimulation of host defenses. Ginekol Pol, 2003 Dec, 74(12), 1526 - 30 {Importance of the vaginal ecosystem examination in the first trimester of pregnancy}; Peterek J; OBJECTIVES: Vaginal ecosystem disturbances especially bacterial vaginosis are strongly associated with infectious complication during the course of pregnancy . DESIGN: The aim of the study was the determination of the vaginal ecosystem examination for the diagnosis of the following infection: bacterial vaginosis, candidiasis and trichomoniasis in pregnant women in the first trimester of pregnancy . MATERIALS AND METHODS: The studies included 106 randomly selected asymptomatic pregnant women in I trimester of pregnancy coming for gynaecological examination for the first time after cessation of menstruation were divided into two group: 56 primiparae and 50 multiparae . The diagnosis of BV was based on Amsel criteria concerning vaginal secretion . Trichomonas vaginalis was determined in wet vaginal smear . Candida albicans was detected in vaginal smear after addition of 10% KOH solution . RESULTS: Out of 106 studied subjects in the first trimester of pregnancy, BV was detected in 17.9%, Candida albicans in 15.1%, Trichomonas vaginalis in 3.8% . The detectability of BV infection varied significantly between both groups of studied women (p < 0.001) while no correlation was found between the occurrence of Candida albicans and Trichomonas vaginalis infections . CONCLUSION: Vaginal ecosystem screening in pregnant women makes possible under ambulatory conditions, to detect the following infections: BV, candidiasis and trichomoniasis . Determination of disturbances in vaginal ecosystem is simple and inexpensive, not requiring laboratory facilities for the assessment of vaginal infections. Eur J Clin Microbiol Infect Dis, 2004 Apr, 23(4), 317 - 22 Epub 2004 Mar 17. Epidemiology of candidaemia in Europe: results of 28-month European Confederation of Medical Mycology (ECMM) hospital-based surveillance study; Tortorano AM et al.; In order to update the epidemiological and mycological profile of candidaemia in Europe, the European Confederation of Medical Mycology conducted a prospective, sequential, hospital population-based study from September 1997 to December 1999 . A total of 2,089 cases were documented by 106 institutions in seven European countries . Rates of candidaemia ranging from 0.20 to 0.38 per 1,000 admissions were reported . Candida albicans was identified in 56% of cases . Non-albicans Candida species were most frequently isolated from patients with haematological malignancies (65%) . With increasing age, an increasing incidence of Candida glabrata was seen . The 30-day mortality rate was 37.9% . The survey results underline the burden of candidaemia in a wide range of patient populations, confirm the importance of non- albicans species, and provide baseline data for future surveillance studies at a European level. Antonie Van Leeuwenhoek, 2004 Jan, 85(1), 63 - 8 Cr(VI) reduction in a chromate-resistant strain of Candida maltosa isolated from the leather industry; Ramirez-Ramirez R et al.; A Cr(VI)-resistant yeast was isolated from tanning liquors from a leather factory in Leon, Guanajuato, Mexico . Based on morphological and physiological analyses and the D1/D2 domain sequence of the 26S rDNA, the yeast was identified as Candida maltosa . Resistance of the strain to high Cr(VI) concentrations and its ability to chemically reduce chromium was studied . When compared to the three laboratory yeasts Candida albicans, Saccharomyces cerevisiae and Yarrowia lipolytica, the C . maltosa strain was found to tolerate chromate concentrations as high as 100 micro g/ml . In addition to this phenotypic trait, the C . maltosa strain showed ability to reduce Cr(VI) . Chromate reduction occurred both in intact cells (grown in culture medium or in soil containing chromate) as well as in cell-free extracts . NADH-dependent chromate reductase activity was found associated with soluble protein and, to a lesser extent, with the membrane fraction. Bioorg Med Chem, 2004 Apr 1, 12(7), 1751 - 68 Mannich reaction: an approach for the synthesis of water soluble mulundocandin analogues; Lal B et al.; Semisynthetic modifications at Hydroxy tyrosine (Htyr) unit of mulundocandin (1) were carried out to improve its aqueous solubility . A single step introduction of substituted aminomethyl groups at the ortho position(s) of phenolic hydroxyl of HTyr unit of mulundocandin has been achieved in 7-85% yield . The in vitro screening of Mannich products against Candida albicans and Aspergillus fumigatus, retained the in vivo activity of parent by oral and intraperitoneal route . Compound 20, showed significant improvement in activity over mulundocandin (1) and activity compares well with that of fluconazole. Indian J Pathol Microbiol, 2003 Jan, 46(1), 124 - 6 Evaluation of germ tube test in various media; Arora DR et al.; One of the most valuable tests for the rapid presumptive identification of Candida albicans is germ tube production . Previously researchers have concluded superior results with serum in germ tube test . However, handling of pooled human serum has danger of acquisition of hepatitis and HIV infections . Therefore, in the present study we compared various media--pooled human serum, sterile horse serum, peptone water and tryptic soy broth . It was observed that human serum remains to be the best with 100% positivity for identification of C . albicans followed by horse serum (76.3%), peptone water (61.8%) and tryptic soy broth (61.8%). J Pharm Pharmacol, 2004 Mar, 56(3), 285 - 9 Histatins: antimicrobial peptides with therapeutic potential; Kavanagh K et al.; Histatins are a group of antimicrobial peptides, found in the saliva of man and some higher primates, which possess antifungal properties . Histatins bind to a receptor on the fungal cell membrane and enter the cytoplasm where they target the mitochondrion . They induce the non-lytic loss of ATP from actively respiring cells, which can induce cell death . In addition, they have been shown to disrupt the cell cycle and lead to the generation of reactive oxygen species . Their mode of action is distinct from those exhibited by the conventional azole and polyene drugs, hence histatins may have applications in controlling drug-resistant fungal infections . The possibility of utilising histatins for the control of fungal infections of the oral cavity is being actively pursued with the antifungal properties of topical histatin preparations and histatin-impregnated denture acrylic being evaluated . Initial clinical studies are encouraging, having demonstrated the safety and efficacy of histatin preparations in blocking the adherence of the yeast Candida albicans to denture acrylic, retarding plaque formation and reducing the severity of gingivitis . Histatins may represent a new generation of antimicrobial compounds for the treatment of oral fungal infections and have the advantage, compared with conventional antifungal agents, of being a normal component of human saliva with no apparent adverse effects on host tissues and having a mode of action distinct to azole and polyene antifungals. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2004 Mar, 97(3), 351 - 8 Evaluation of the recurrence of denture stomatitis and Candida colonization in a small group of patients who received itraconazole; Cross LJ et al.; OBJECTIVES: The aim of the study was to determine the recurrence rate of denture stomatitis and persistence of Candida in 22 patients (5 male and 17 female, mean age 71 years) over a 3-year period . STUDY DESIGN: Denture hygiene practice, denture cleanliness, and the presence of palatal erythema were assessed for each patient at the start of the study (baseline) . The oral cavity was sampled for yeasts by imprint culture and denture discs . Ten patients received a capsular form of itraconazole (100 mg twice daily for 15 days) and 12 patients were provided with 100 mg of itraconazole in the form of a mouthwash (10 mL twice daily), which was then swallowed . No further antifungal treatment was administered to any of the patients . Clinical and microbiological assessments were repeated for each patient at 6 months and 3 years after the original appointment . Yeasts were identified by colony color on CHROMagar Candida, germ-tube formation, and API-32C profiling . Selected isolates were then typed by inter-repeat polymerase chain reaction (IR PCR) . RESULTS: Candida albicans was isolated at baseline from all patients either alone (12 patients) or in combination with another species (10 patients) . Other yeast species recovered were C glabrata (5 patients), C tropicalis (1 patient), C guilliermondii (1 patient), C krusei (1 patient), C parapsilosis (1 patient), C kefyr (1 patient), and Saccharomyces cerevisiae (2 patients) . Candida albicans and/or C glabrata were recovered from 11 of the 22 patients after 6 months or 3 years . A complete and consistent change of yeast species from baseline was observed in 6 patients after 6 months and at 3 years . The remaining 5 patients were yeast-free at the follow-up assessments . PCR fingerprinting of C albicans and C glabrata indicated strain persistence over 6 months in 10 patients and in 4 patients after 3 years . A switch in strain type occurred for 1 patient after 6 months and for 3 patients after 3 years . CONCLUSIONS: The recurrence of denture stomatitis in patients who maintained a high standard of denture cleanliness was low . Although itraconazole was beneficial in reducing the fungal load, there may be strain persistence or subsequent recolonization of the oral cavity by a broader range of potentially less sensitive yeast species. Diagn Microbiol Infect Dis, 2004 Mar, 48(3), 153 - 60 The role of antifungal susceptibility testing in the therapy of candidiasis; Hospenthal DR et al.; Prior to the introduction of azoles, no real need for antifungal susceptibility testing (AFST) existed, as amphotericin B was the only agent available to treat systemic candidiasis . Introduction of fluconazole and itraconazole provided alternate, less toxic antifungal therapies . Intrinsic resistance of Candida krusei, decreased susceptibility of Candida glabrata, and development of resistance by Candida albicans (in mucosal disease in AIDS) to azoles led to development of our current AFST methodologies . The goal of AFST, like that of antibacterial susceptibility testing, is to predict clinical response, or at least to forecast failure . Although the ability of AFST to predict clinical outcome (clinical correlation) is still being fully elucidated, current methodologies do appear to reliably predict clinical resistance to azoles . Ready access to AFST is currently limited, affecting its timely use, but even with this lack of timeliness, AFST can still play an important role in patient care . Important potential roles include: 1) use in the development of local antibiograms to aid empiric selection of antifungals; 2) testing of isolates from candidemia or deep infection to aid in selection of long-term therapies; and, 3) the testing of isolates from recurrent mucosal disease to aid in selection of alternative regimens. Inflamm Res, 2004 Feb, 53(2), 72 - 7 Epub 2004 Jan 26. Histopathological features of murine systemic vasculitis caused by Candida albicans extract--an animal model of Kawasaki disease; Takahashi K et al.; OBJECTIVE AND DESIGN: We examined the histopathological features of systemic vasculitis caused in mice by injection of a Candida albicans ( C . albicans) extract and investigated the principal genetic roles in the development of vasculitis . MATERIALS AND METHODS: C . albicans extract was injected intraperitoneally for five consecutive days in the 1st and 5th weeks to CD-1, C57BL/6N, C3H/HeN, BALB/cAnN, DBA/2N and CBA/JN mice . At week 8, mice were killed, and histological examination was performed by light microscopy . RESULTS: Arteritis had developed in 66% of CD-1 mice . The extramural coronary arteries and aortic root close to the orifice of coronary arteries were most frequently involved . Histologically, the characteristic feature of the arteritis was proliferative and granulomatous inflammation accompanied by numerous macrophages, lymphocytes, plasma cells and neutrophils . Fibrocellular intimal thickening with destruction of the internal elastic lamina and media was also observed . Five mouse strains after injection of C . albicans extract were clearly classified into a resistant group (CBA/JN, DBA/2N and BALB/cAnN mice) and a sensitive group (C3H/HeN and C57BL/6N mice) . The inbred mouse strains which showed the same histocompatibility-2 (H-2) haplotype exhibited a different susceptibility to development of vasculitis . CONCLUSION: This arteritis murine model shows unique histological features that have not been observed in other animal vasculitis models and it most closely resembles Kawasaki disease in humans . The genetic control of susceptibility to induction of vasculitis by the C . albicans extract is dependent to the mouse strains, but is not linked to the H-2 loci. Peptides, 2003 Nov, 24(11), 1807 - 14 Prevention of lethal murine candidiasis using HP (2-20), an antimicrobial peptide derived from the N-terminus of Helicobacter pylori ribosomal protein L1; Ribeiro PD et al.; Peptide HP (2-20), A(2)KKVFKRLEKLFSKIQNDK(20), is a cationic antimicrobial peptide derived from the N-terminus of Helicobacter pylori ribosomal protein 1, HpRpL1 . Native peptide HP (2-20) and its synthetic derivatives have been shown in vitro to exhibit potent killing activity against Gram-positive, Gram-negative and yeast cells, thus, making them promising candidates for treatment of polymicrobial infections . However, the therapeutic potential of peptide HP (2-20) or its synthetic derivatives in any animal model of either bacterial or fungal diseases has not yet been investigated . In this study, we demonstrate that synthetic peptide amide HP (2-20), administered in six doses (300microg each; one intraperitoneal dose at the time of the infection, followed by five intravenous doses at 12h intervals) to CBA/J male mice experimentally infected with a lethal inoculum ( {Formula: see text} CFU) of Candida albicans, delayed the onset of disease, suppressed disease progression, and greatly increased survival rate and time (16.6% by day 14), as compared with the untreated infected control mice (100% mortality by day 5) . Further, using isotonic buffer systems differing in ionic strength, peptide HP (2-20) was shown in vitro to exhibit an ionic strength-dependent hemolytic activity, previously not detected . Repeated intravenous administration of uninfected control CBA/J male mice with peptide HP (2-20), however, caused neither morbidity nor mortality . These findings strongly evidence the therapeutic efficacy and safety values of peptide HP (2-20) as a lead drug for the treatment of acquired candidiasis. J Pharm Biomed Anal, 2004 Mar 10, 34(5), 1117 - 24 Liquid chromatography-tandem mass spectrometric method for the analysis of fluconazole and evaluation of the impact of phenolic compounds on the concentration of fluconazole in Candida albicans; Sun S et al.; A bioanalytical method using liquid chromatography-tandem mass spectrometry was developed for the analysis of fluconazole in Candida albicans after incubation with phenolic compounds, which have been proved possessing antifungal properties and have synergetic activity against C . albicans when in combination with fluconazole . Samples of C . albicans thallus obtained by centrifuging the mixed culture after 24 h incubation were saponified and centrifuged . The supernatant was evaporated to dryness, reconstituted, and injected on a C18 column using an organic-aqueous mobile phase . The chromatographic run time was 3.5 min per injection, with retention times of 2.4 min for fluconazole . The detection was by monitoring fluconazole at m/z 305-->191 . The standard curve range was 1.0-100.0 ng ml(-1) with a mean correlation coefficient 0.9992 . The precision and accuracy of the quality control (QC) samples were R.S.D < 5.5%, R.E . <3% for intra-day and R.S.D . <6.2%, R.E . <4% for inter-day . The concentration of fluconazole in C . albicans was found to be increased with the increment of the tested compounds concentration when they were in combination. Clin Diagn Lab Immunol, 2004 Mar, 11(2), 344 - 50 Circulating beta (1-3) glucan and immunoglobulin G subclass antibodies to Candida albicans cell wall antigens in patients with systemic candidiasis; Kondori N et al.; Invasive candidiasis in patients who are immunocompromised or in intensive care units (ICUs) presents both diagnostic and therapeutic problems . We previously described antibodies that were directed against Candida albicans cell wall fragments (CW), periodate-treated CW (CW(IO4)), phosphopeptidomannan (PPM), and beta(1-3) glucan . In this study, circulating fungal antigens {mannan and beta(1-3) glucan} and immunoglobulin G (IgG) subclass antibodies to these cell wall antigens (anti-CW) were analyzed in patients with systemic candidiasis . Sera were collected from 14 patients on two or three consecutive occasions, starting on the day when candidiasis was culture proven . The sera were analyzed by enzyme-linked immunosorbent assay . The control groups consisted of lactating mothers (n = 9) (group I) who had breast milk that was positive for C . albicans and also had acute inflammation of the nipples, and age-matched blood donors (n = 10) (group II) . Within the first 3 weeks of Candida infection all of the patients were positive for beta(1-3) glucan by the Gluspecy test, but no patients were positive for mannan in the less-sensitive Pastorex Candida test . The controls were negative for both beta(1-3) glucan (<20 pg/ml) and mannan (<2.5 ng/ml) . IgG1 anti-CW and IgG2 anti-PPM antibodies were the most discriminatory antibodies . The ratio of IgG1 anti-CW to IgG2 anti-PPM was significantly lower in nonsurviving patients than in the other patients within the first week of candidiasis (P = 0.019) . The IgG2 levels of anti-CW(IO4) and antiglucan antibodies correlated strongly (r = 0.681; P < 0.0001), and the absence of these antibodies was associated with increased levels of beta(1-3) glucan . Increased levels of IgG1 anti-CW or IgG2 anti-PPM antibodies (titer of > or = 3 logs) or of a combination of the two antibodies (log sum, > or = 5) showed 92% sensitivity, 100% specificity, and positive predictive values . In conclusion, beta(1-3) glucan and the two subclass antibodies appear to be early specific markers for the laboratory diagnosis of candidiasis . Furthermore, the kinetics of beta(1-3) glucan appearance in serum may assist in evaluating the therapeutic efficacy of antifungal treatments. FEBS Lett, 2004 Mar 12, 561(1-3), 223 - 30 Candida albicans Cdc37 interacts with the Crk1 kinase and is required for Crk1 production; Ni J et al.; Crk1, a Cdc2-related protein kinase from the human pathogenic fungus Candida albicans, plays an important role in hyphal development and virulence . To address its regulatory mechanisms, we searched for Crk1 interacting proteins by two-hybrid screening . A CDC37 ortholog (CaCDC37) was cloned from the screening with the Crk1 kinase domain as the bait . The CaCdc37 interacted preferentially with the kinase domain of Crk1 (Crk1N) as shown by two-hybrid and immunoprecipitation experiments . CaCDC37 could complement a cdc37 thermosensitive mutant (cdc37-34) of Saccharomyces cerevisiae . Importantly, Crk1 protein was hardly detectable in the cdc37-34 mutant at restrictive temperature . However, upon expression of CaCdc37 in the cdc37 mutant, Crk1 protein was detected even at restrictive temperature . Our data suggested that CaCdc37 was required for the production of Crk1 kinase . Like Cdc37 proteins of S . cerevisiae and higher eukaryotes, CaCdc37 might function as a molecular chaperone that stabilized Crk1 and other protein kinases in C . albicans . In support of this, CaSTI1 was identified from a two-hybrid screen with the full-length Crk1 as the bait . CaSti1 showed two-hybrid interactions with both Crk1 and the CaCdc37. J Appl Microbiol, 2004, 96(4), 803 - 9 Quantitative evaluation of antifungal activity of metallic oxide powders (MgO, CaO and ZnO) by an indirect conductimetric assay; Sawai J et al.; AIMS: To evaluate antifungal activities of MgO, CaO and ZnO powders quantitatively by indirect conductimetric assay . METHODS AND RESULTS: Candida albicans NBRC1060, Saccharomyces cerevisiae NBRC1950, Aspergillus niger NBRC4067 and Rhizopus stolonifer NBRC4781 were used as test micro-organisms . The indirect conductimetric assay, in which the change in electrical conductivity of an alkaline solution (NaOH) is produced by absorption of CO2 from microbial metabolism, could offer a simple and rapid evaluation of the antifungal activity within 24-48 h . The conductivity curves obtained for MgO, CaO and ZnO were analysed using the growth inhibition kinetic model proposed by Takahashi for calorimetric evaluation, and the kinetic parameters and minimum inhibitory concentration ({I}100) could be determined . MgO and CaO powders exhibited the antimicrobial activities against all fungi used in this study and showed little differences between types of fungi . However, although ZnO powder inhibited fungal growth, the values of {I}100 were over 100 mg ml-1 . CONCLUSIONS: Although a common method for evaluating antifungal activity requires over 5-7 days, the indirect assay could provide a rapid and quantitative evaluation of antifungal activity within approx . 2 days, and MgO and CaO were found to have antifungal activities . SIGNIFICANCE AND IMPACT OF THE STUDY: The indirect assay can be applicable for simple and rapid evaluation of the antimicrobial activity of insoluble or slightly soluble materials with high turbidity such as antibacterial ceramic powders . Moreover, these materials can be useful for controlling fungi in food processing and the environment. Immunology, 2004 Mar, 111(3), 334 - 42 Protection against systemic candidiasis in mice immunized with secreted aspartic proteinase 2; Vilanova M et al.; Secreted aspartic proteinases (Sap) have been described as virulence factors implicated in the mechanisms of host colonization by the yeast Candida albicans in different types of candidiasis . Intraperitoneal inoculation of C . albicans into BALB/c mice rapidly leads to systemic candidiasis, with significant colonization of the kidneys measurable in the following week . In this study we assessed the potential of vaccination with C . albicans secreted aspartic proteinase 2 (Sap2) in preventing systemic candidiasis in BALB/c mice . Intradermal injection of highly purified native Sap2 protein incorporated in alum adjuvant provided efficient immune protection, as indicated by a 20-fold decrease in the colonization of kidneys . The protective effect of Sap2 immunization with alum adjuvant was also observed in mice infected with a lethal inoculum of C . albicans . Immunization with the native Sap2 alone, as well as with a denatured recombinant form of the protein, also conferred protection, albeit to a lesser level . In all cases, protection correlated with an increase in serum antibodies to Sap2 . Moreover, passive transfer of anti-Sap2 immunoglobulin G (IgG) significantly decreased the yeast burden in kidneys of C . albicans-infected mice . This result shows that immune protection against systemic candidiasis in mice immunized with Sap2 is antibody-mediated . Taken together, these analyses demonstrate that Sap2 can be successfully used as a vaccination target in systemic candidiasis and reveals the potential immunomodulatory role of Sap2 on C . albicans infection. Eur J Biochem, 2004 Mar, 271(6), 1219 - 26 Antimicrobial activities of heparin-binding peptides; Andersson E et al.; Antimicrobial peptides are effector molecules of the innate immune system . We recently showed that the human antimicrobial peptides alpha-defensin and LL-37 bind to glycosaminoglycans (heparin and dermatan sulphate) . Here we demonstrate the obverse, i.e . structural motifs associated with heparin affinity (cationicity, amphipaticity, and consensus regions) may confer antimicrobial properties to a given peptide . Thus, heparin-binding peptides derived from laminin isoforms, von Willebrand factor, vitronectin, protein C inhibitor, and fibronectin, exerted antimicrobial activities against Gram-positive and Gram-negative bacteria . Similar results were obtained using heparin-binding peptides derived from complement factor C3 as well as consensus sequences for heparin-binding (Cardin and Weintraub motifs) . These sequence motifs, and additional peptides, also killed the fungus Candida albicans . These data will have implications for the search for novel antimicrobial peptides and utilization of heparin-protein interactions should be helpful in the identification and purification of novel antimicrobial peptides from complex biological mixtures . Finally, consensus regions may serve as templates for de novo synthesis of novel antimicrobial molecules. Appl Environ Microbiol, 2004 Mar, 70(3), 1356 - 9 Inoculum size effect in dimorphic fungi: extracellular control of yeast-mycelium dimorphism in Ceratocystis ulmi; Hornby JM et al.; We studied the inoculum size effect in Ceratocystis ulmi, the dimorphic fungus that causes Dutch elm disease . In a defined glucose-proline-salts medium, cells develop as budding yeasts when inoculated at > or = 10(6) spores per ml and as mycelia when inoculated at <10(6) spores per ml . The inoculum size effect was not influenced by inoculum spore type, age of the spores, temperature, pH, oxygen availability, trace metals, sulfur source, phosphorous source, or the concentration of glucose or proline . Similarly, it was not influenced by added adenosine, reducing agents, methyl donors, amino sugars, fatty acids, or carbon dioxide . Instead, growing cells excreted an unknown quorum-sensing factor that caused a morphological shift from mycelia to budding yeasts . This yeast-promoting effect is abolished if it is extracted with an organic solvent such as ethyl acetate . The quorum-sensing activity acquired by the organic solvent could be added back to fresh medium in a dose-dependent fashion . The quorum-sensing activity in C . ulmi spent medium was specific for C . ulmi and had no effect on the dimorphic fungus Candida albicans or the photomorphogenic fungus Penicillium isariaeforme . In addition, farnesol, the quorum-sensing molecule produced by C . albicans, did not inhibit mycelial development of C . ulmi when present at concentrations of up to 100 microM . We conclude that the inoculum size effect is a manifestation of a quorum-sensing system that is mediated by an excreted extracellular molecule, and we suggest that quorum sensing is a general phenomenon in dimorphic fungi. J Immunol, 2004 Mar 15, 172(6), 3712 - 8 Toll-like receptor 2 suppresses immunity against Candida albicans through induction of IL-10 and regulatory T cells; Netea MG et al.; Toll-like receptor (TLR) 2 and TLR4 play a pivotal role in recognition of Candida albicans . We demonstrate that TLR2(-/-) mice are more resistant to disseminated Candida infection, and this is associated with increased chemotaxis and enhanced candidacidal capacity of TLR2(-/-) macrophages . Although production of the proinflammatory cytokines TNF, IL-1alpha, and IL-1beta is normal, IL-10 release is severely impaired in the TLR2(-/-) mice . This is accompanied by a 50% decrease in the CD4+CD25+ regulatory T (Treg) cell population in TLR2(-/-) mice . In vitro studies confirmed that enhanced survival of Treg cells was induced by TLR2 agonists . The deleterious role of Treg cells on the innate immune response during disseminated candidiasis was underscored by the improved resistance to this infection after depletion of Treg cells . In conclusion, C . albicans induces immunosuppression through TLR2-derived signals that mediate increased IL-10 production and survival of Treg cells . This represents a novel mechanism in the pathogenesis of fungal infections. J Clin Microbiol, 2004 Mar, 42(3), 1288 - 9 Stability of Mueller-Hinton agar supplemented with glucose and methylene blue for disk diffusion testing of fluconazole and voriconazole; Pfaller MA et al.; The shelf life of Mueller-Hinton agar supplemented with 2% glucose and methylene blue (0.5 micro g/ml) (MH-GMB) prepared in the laboratory to test disk diffusion of voriconazole and fluconazole was assessed using quality control (QC) strains of Candida albicans ATCC 90028, Candida krusei ATCC 6258, and Candida parapsilosis ATCC 22019 . MH-GMB agar plates were prepared as described in National Committee for Clinical Laboratory Standards document M44-P, and isolates were tested using 25- micro g fluconazole disks and 1- micro g voriconazole disks over a 36-day period . Zone diameters for fluconazole and voriconazole varied by no more than 4 mm over the study period, and 95 to 100% of results were within the established QC limits for the strains tested . Prepared MH-GMB agar plates provide acceptable performance for disk diffusion testing for at least 30 days when stored at 5 degrees C. Microb Pathog, 2004 Apr, 36(4), 211 - 7 Bone marrow colony-formation in vitro after infection of genetically defined inbred mice with Candida albicans; Wanasaengsakul S et al.; The effect of C . albicans infection on the production of haematopoietic precursor cells in the bone marrow of CBA/CaH and BALB/c mice was evaluated by assay of colony formation in vitro . In immunocompetent mice, neither systemic nor oral infection induced significant alterations in colony formation by bone marrow from the two mouse strains, and Candida infection did not alter the proportion of morphological cell types in the colonies . However, the number of neutrophil-like was relatively greater in colonies derived from acutely infected CBA/CaH nude mice than in those from BALB/c nude mice, whereas small mononuclear cells were present in higher proportions in the latter strain . In both strains of nude mice, there was an increase in colony formation at 6 days after oral infection, but at 8 weeks, when the infection had become chronic, the production of bone marrow cells by CBA/CaH nude mice was significantly less than that by BALB/c nude mice . Reconstitution of nude mice with syngeneic lymphocytes enhanced the production of bone marrow precursor cells by BALB/c, but not by CBA/CaH mice, suggesting that T cells can enhance host resistance by promoting the colony-forming response of the bone marrow in BALB/c mice that are genetically resistant to tissue damage, but not in CBA/CaH that are prone to severe lesions . Finally, culture with Candida antigen in vitro decreased the number of colony-forming cells in cultures from CBA/CaH, but not from BALB/c mice. Scand J Infect Dis, 2004, 36(1), 52 - 5 A prospective epidemiological survey of candidaemia in Sweden; Klingspor L et al.; A prospective epidemiological survey of candidaemia was performed in central Sweden from January 1998 to December 1999 . In total, 191 episodes were reported with an overall rate of 0.32/1000 admissions . Candida albicans was identified in 128 cases (67%), followed by Candida glabrata in 30 (15.7%) and Candida parapsilosis in 14 (7.3%) . Predisposing factors included surgery (31.4%), intensive care (18.8%), solid tumour or haematological malignancy (15.7%), and foetal immaturity (15.7%) . Non-albicans Candida species were more prevalent among patients with haematological malignancies (56%), compared to surgical (30%) and ICU patients (19%) . The crude mortality rate of candidaemia was 31% . The highest mortality rate was observed in patients with haematological malignancies (41.2%), age > 70 y (41%), surgery (38.5%) and infections with > 1 Candida species (40%) or C . glabrata (38%). Laryngorhinootologie, 2004 Feb, 83(2), 117 - 21 {From when on can fungi be identified in nasal mucus of humans?}; Lackner A et al.; BACKGROUND: Fungal spores are frequent in air and their occurrence in the nasal mucus appears to be a common finding within the adult population, as we were able to show in recent studies . 91,3 % of CRS patients but also healthy controls grew positive fungal cultures out of their nasal mucus . The potential role of fungal elements in nasal mucus for the pathogenesis of CRS, with or without polyposis, is currently investigated intensely and discussed very controversially . However, it was still unknown, as of when fungi could be cultured from nasal mucus in humans . We attempted to identify this point of time, in the nasal mucus of neonates . METHODS: In our study we examined nasal mucus from 30 neonates immediately after birth, on the first and fourth day post partum, and after two and four months of life . The samples obtained with sterile cotton swabs were cultured on agar plates . Fungal cultures were identified either conventionally by microscopy or with molecular techniques . To show whether fungi in nasal mucus of newborns were acquired by contamination during birth, mucus of the maternal vagina was examined as well . RESULTS: Just after birth we found in 6 of 30 (20 %) of our neonates positive fungal cultures out of their nasal mucus, in 3 of them Candida albicans, probably due to contamination passing the maternal vagina as cultures of vaginal mucus of their mothers were positive for Candida albicans too . Positive fungal cultures were obtained in 2 of 29 (7 %) neonates on the second and in 4 of 26 (15 %) neonates on the fifth day of life . In all our cases initial presence in nasal mucus contamination just after birth or on the second day of life was limited to one day only . None of the 12 of 30 (40 %) neonates with positive fungal cultures from nasal mucus in the first 5 days of life showed clinical symptoms of nasal fungal colonisation . Besides Candida albicans, Penicillium sp., Cladosporium cladosporioides, Acremonium polychromum, Beauveria bassiana and Epicoccum nigrum could be detected in the first 5 days of life . After the second month of life, examination of nasal mucus yielded positive fungal cultures in 8 of 11 (72 %), after four months even 17 of 18 (94 %) of babies, with a wide array of different species . CONCLUSIONS: Fungi can be cultured from nasal mucus as soon as contact with the environmental air exists . Furthermore, a transfer of fungi from the mother's birth canal into the nose during birth is possible . Presence of fungal spores is common but not persistent in the nose of babies in the first days of life . However, after four months the situation is similar to the one in adults: fungal cultures can be obtained from almost everyone's nose . Therefore fungal spores must be considered a normal content of nasal mucus . Fungal spores are inhaled with every breath, some stick to the mucus, are transported to the nasopharynx and swallowed . This does not cause any clinical symptoms and is therefore not a pathological finding at all. Mycoses, 2004 Feb, 47(1-2), 24 - 8 Phylogenetic relationships among medically important yeasts based on sequences of mitochondrial large subunit ribosomal RNA gene; Yamada Y et al.; Sequences of the mitochondrial large subunit ribosomal RNA (mtLsurRNA) gene of medically important yeasts were analysed . Sixteen strains of eight species including two varieties were subjected to sequencing . Sequencing enabled us to recognize the differences between all the species and varieties . Alignment analysis revealed that these sequences consisted of three clusters: the Candida albicans group, the C . glabrata group, and the basidiomycetous group . It is possible, therefore, that the mtLsurRNA gene is one of the targets not only for species identification but also for phylogenetic analysis of closely related yeasts . The dendrogram of each group, obtained from this gene, supports the previous study of yeasts based upon the chromosomal genes. J Med Chem, 2004 Mar 11, 47(6), 1553 - 74 Design, synthesis, and evaluation of aza-peptide epoxides as selective and potent inhibitors of caspases-1, -3, -6, and -8; James KE et al.; Aza-peptide epoxides, a novel class of irreversible protease inhibitors, are specific for the clan CD cysteine proteases . Aza-peptide epoxides with an aza-Asp residue at P1 are excellent irreversible inhibitors of caspases-1, -3, -6, and -8 with second-order inhibition rates up to 1 910 000 M(-1) s(-1) . In general, the order of reactivity of aza-peptide epoxides is S,S > R,R > trans > cis . Interestingly, some of the R,R epoxides while being less potent are actually more selective than the S,S epoxides . Our aza-peptide epoxides designed for caspases are stable, potent, and specific inhibitors, as they show little to no inhibition of other proteases such as the aspartyl proteases porcine pepsin, human cathepsin D, plasmepsin 2 from P . falciparum, HIV-1 protease, and the secreted aspartic proteinase 2 (SAP-2) from Candida albicans; the serine proteases granzyme B and alpha-chymotrypsin; and the cysteine proteases cathepsin B and papain (clan CA), and legumain (clan CD). J Biol Chem, 2004 May 7, 279(19), 20250 - 6 Epub 2004 Mar 02. Identification and characterization of a mucosal antimicrobial peptide expressed by the chinchilla (Chinchilla lanigera) airway; Harris RH et al.; Cationic antimicrobial peptides (APs) are produced at mucosal surfaces and play a key role as a first line of defense against infection . To understand how APs might impact disease progression in otitis media (OM), our goal was to identify and characterize APs expressed by the epithelium lining the uppermost airway of the chinchilla, the established rodent host for the study of the bacterial-viral pathogenesis in OM . Using a molecular approach, we cloned a cDNA encoding a homolog of human beta-defensin 3, designated chinchilla beta-defensin-1 (cBD-1), and found by Northern analysis expression of the corresponding mRNA in nasopharyngeal and tongue mucosae as well as skin . By reverse transcription-PCR, cBD-1 mRNA was also detected in RNA isolated from trachea, lung, and Eustachian tube tissues . The predicted mature form of cBD-1, expressed as a recombinant peptide in Escherichia coli, demonstrated bactericidal activity against the three primary opportunistic pathogens of OM as well as Candida albicans . Continued study of this and other APs will allow us to determine their role in bacterial colonization of the upper airway as well as how viruses might contribute to the pathogenesis of OM by modulating AP expression. Oral Dis, 2004 Mar, 10(2), 81 - 6 Inhibition of adherence of Candida albicans and Candida dubliniensis to a resin composite restorative dental material by salivary secretory IgA and monoclonal antibodies; Elguezabal N et al.; OBJECTIVE: The attachment of Candida to oral surfaces is a crucial step in the colonization of the oral cavity and the eventual development of oral diseases caused by this microorganism . Inhibition of adhesion is one of the strategies currently studied to prevent Candida infections . The main objective of this study was to investigate the inhibitory effect of the human salivary components on the adherence of Candida albicans and C . dubliniensis to Herculite, a widely used resin composite restorative dental material . We have also investigated the influence on the adherence of three monoclonal antibodies (mAbs) directed against C . albicans cell wall antigens . DESIGN: The adhesion of three strains of C . albicans and one strain of C . dubliniensis was studied by a visual method after incubating the fungus and the resin in presence and in absence of human whole saliva, secretory immunoglobulin A (sIgA) and three mAbs directed against C . albicans cell wall surface antigens . RESULTS: Adherence of C . albicans was inhibited by whole saliva (41.7%), salivary sIgA (55.7%) and the salivary components that bind to the cell wall (36.7%) . Whole saliva significantly reduced the adhesion of C . dubliniensis to Herculite to 45.3% of the control level . Saliva previously adsorbed with fungal cells or sIgA depleted saliva had no effect on adherence . An inhibition in the adhesion of C . albicans and C . dubliniensis to Herculite similar to that shown by whole saliva was also observed when mAbs C7 and 26G7 were used . However, mAb 21E6 increased adhesion of all the strains to Herculite . CONCLUSIONS: The results suggest that sIgA, as well as whole saliva, are important in blocking adherence of C . albicans and C . dubliniensis to Herculite and that this effect can be reproduced with mAbs directed against the cell wall surface of C . albicans. J Mass Spectrom, 2004 Feb, 39(2), 193 - 201 De novo sequencing of antimicrobial peptides isolated from the venom glands of the wolf spider Lycosa singoriensis; Budnik BA et al.; Antimicrobial peptides (AMPs), named lycocitin 1, 2 and 3, and a peptide with a monoisotopic molecular mass of 3038.70 Da were detected in the venom glands of the wolf spider Lycosa singoriensis . Two of the peptides, lycocitin 1 and 2, are new AMPs whereas lycocitin 3 is highly homologous to lycotoxin II isolated from the venom of spider Lycosa carolinensis . In addition, two other peptides with monoisotopic masses of 2034.20 and 2340.28 Da showing the motif typical for antimicrobial peptides were also identified . These peptides and lycocitin 1, 2 and 3 were de novo sequenced using electron capture dissociation and low-energy collisional tandem mass spectrometry . The amino acid sequence of lycocitin 1 was determined as GKLQAFLAKMKEIAAQTL-NH(2) . Lycocitin 2 differs from lycocitin 1 by a replacement of a lysine residue for an arginine residue at the second position . Lycocitin 3 differs from the known lycotoxin II consisting of 27 amino acid residues by a deletion of Gly-26 . Both lycocitin 1 and 2 inhibit growth of Gram-positive (Staphylococcus aureus, Bacillus subtilis) and Gram-negative (Escherichia coli) bacteria and fungi (Candida albicans, Pseudomonas aeruginosa) at micromolar concentrations . J Infect Chemother, 2004 Feb, 10(1), 59 - 61 Reduced activity of Candida detachment factors in the saliva of the elderly; Kamagata-Kiyoura Y et al.; Saliva in healthy human subjects contains protenaceous factors that detach Candida albicans cells from plastic plates . To investigate the physiological role of the detachment activity of human saliva, we compared the activity of salivary specimens collected from the elderly (who are at greater risk of oral candidiasis) to the activity of specimens collected from children and nonelderly adults . C . albicans cells were cultured and attached to the bottoms of the wells of a 96-well plate, and saliva preparations were then added to each well . After 3 h of incubation, Candida cells attached to the bottoms of the wells were counted, using the crystal violet staining method . The results showed that the activity of salivary detachment factors was significantly lower among the elderly than among nonelderly adults and children . Furthermore, the activity of salivary detachment factors was reduced in 12 of the 43 elderly subjects (27.9%), a significantly higher proportion relative to that in the children and nonelderly adults . However, there was no significant difference in the level of salivary proteins between the samples collected from elderly subjects displaying low and high salivary detachment activity . These results suggest that the increased number of people with lower salivary detachment activity among the elderly may be one of the reasons that oral candidiasis is more common among the elderly than in nonelderly adults and children. Arch Soc Esp Oftalmol, 2004 Feb, 79(2), 89 - 92 {Recurrence of bacterial endophthalmitis after penetrating keratoplasty}; Garcia Serrano JL et al.; OBJECTIVE/METHODS: One month after penetrating keratoplasty, a male patient suffered recurrent abscess and endophthalmitis caused by mixed bacteria (Staphylococcus aureus and Candida Albicans) . RESULTS: Two samples of vitreous and aqueous were taken, and a combination of systemic Vancomycin and Amphotericin B were injected intravitreally . The patient was asymptomatic for a month, but the abscess reactivated and was treated with Vancomycin and Ciprofloxacin administered intravenously . Fifteen days after withdrawing the treatment endophthalmitis returned and was treated with vitrectomy and additional systemic/intravitreal antibiotics and antifungal therapy . The final visual acuity was 20/40 . CONCLUSIONS: The delayed vitrectomy proved to be necessary to remove established germs and to eliminate recurrent infections. J Am Acad Dermatol, 2004 Mar, 50(3), 343 - 8 Innate immune defense of the nail unit by antimicrobial peptides; Dorschner RA et al.; BACKGROUND: The nail is susceptible to microbial invasion, yet is usually able to defend itself from infection . This occurs despite isolation from cell-mediated immunity . OBJECTIVE: The aim of this study was to determine whether soluble innate immune molecules are present in the nail environment that can protect against microbial colonization . METHODS: Chromatographic techniques were used to purify cationic antimicrobial molecules from porcine hoof extracts . Sections of human and mouse digits were immunostained with antibodies to each species' cathelicidin antimicrobial peptide . Liquid antimicrobial assays were used to determine the activity of these molecules against relevant pathogens . RESULTS: Human, porcine, and murine nails contain antimicrobial molecules, and the human cathelicidin LL-37 can kill Candida albicans . CONCLUSION: The presence of antimicrobial peptides in nails with activity against relevant nail pathogens may account for the ability of the nail unit to resist infection in the absence of direct access to the cellular immune system. Support Care Cancer, 2004 Jul, 12(7), 511 - 6 Epub 2004 Feb 19. High-dose fluconazole therapy for cancer patients with solid tumors and candidemia: an observational, noncomparative retrospective study; Torres HA et al.; BACKGROUND: Response rates for candidemia treated with standard-dose fluconazole (400 mg/day) are approximately 70% . Higher doses of fluconazole have been recommended for susceptible dose-dependent Candida isolates . Herein, we describe the outcome of 20 patients with solid tumors and candidemia treated with high-dose fluconazole (HDF) at The University of Texas M.D . Anderson Cancer Center (1998-2002) . PATIENTS AND METHODS: Patients were identified either by searching the microbiology laboratory database or through direct referral from primary oncology services to the Infectious Diseases Consultative Services . A retrospective review of cases was performed . HDF was defined as > or =600 mg/day . RESULTS: Five patients were treated with 600 mg/day, whereas 15 patients received 800 mg/day . Only one patient was neutropenic . The median APACHE II score at the onset of candidemia was 12 (range 6-24) . The most common species identified were Candida albicans (eight patients, 40%) and Candida parapsilosis (seven patients, 35%) . Of 19 patients whose quantitative data were available, eight (42%) had high-grade candidemia {> or =200 colony forming units (CFU)/ml} . Fifteen (83%) of 18 isolates were fluconazole susceptible, and two (both Candida glabrata) were fluconazole resistant (MIC 64 each) in vitro . Nineteen patients (95%) responded to HDF therapy . The only HDF failure occurred in a patient with C . glabrata (MIC 64.0) infection . The other patient with C . glabrata (MIC 64.0) infection responded to HDF . Central venous catheters were removed from all patients with > or =10 CFU/ml candidemias . All patients with high-grade candidemias responded to HDF . The median duration of HDF therapy was 16 (range 6-42) days . No significant toxicity occurred . CONCLUSIONS: Although our data are limited, HDF appears to be well tolerated and may be associated with higher response rates than standard-dose fluconazole in a selected group of patients with solid tumors and candidemia caused by species that are susceptible to this triazole. Protein Sci, 2004 Mar, 13(3), 703 - 13 Lead optimization of antifungal peptides with 3D NMR structures analysis; Landon C et al.; Antimicrobial peptides are key components of the innate immune response in most multicellular organisms . These molecules are considered as one of the most innovative class of anti-infective agents that have been discovered over the last two decades, and therefore, as a source of inspiration for novel drug design . Insect cystine-rich antimicrobial peptides with the CS alpha beta scaffold (an alpha-helix linked to a beta-sheet by two disulfide bridges) represent particularly attractive templates for the development of systemic agents owing to their remarkable resistance to protease degradation . We have selected heliomicin, a broad spectrum antifungal CS alpha beta peptide from Lepidoptera as the starting point of a lead optimization program based on phylogenic exploration and fine tuned mutagenesis . We report here the characterization, biological activity, and 3D structure of heliomicin improved analogs, namely the peptides ARD1, ETD-135, and ETD-151 . The ARD1 peptide was initially purified from the immune hemolymph of the caterpillars of Archeoprepona demophoon . Although it differs from heliomicin by only two residues, it was found to be more active against the human pathogens Aspergillus fumigatus and Candida albicans . The peptides ETD-135 and ETD-151 were engineered by site-directed mutagenesis of ARD1 in either cationic or hydrophobic regions . ETD-135 and ETD-151 demonstrated an improved antifungal activity over the native peptides, heliomicin and ARD1 . A comparative analysis of the 3D structure of the four molecules highlighted the direct impact of the modification of the amphipathic properties on the molecule potency . In addition, it allowed to characterize an optimal organization of cationic and hydrophobic regions to achieve best antifungal activity. J Immunol, 2004 Mar 1, 172(5), 3070 - 7 Postsecretory processing generates multiple cathelicidins for enhanced topical antimicrobial defense; Murakami M et al.; The production of antimicrobial peptides and proteins is essential for defense against infection . Many of the known human antimicrobial peptides are multifunctional, with stimulatory activities such as chemotaxis while simultaneously acting as natural antibiotics . In humans, eccrine appendages express DCD and CAMP, genes encoding proteins processed into the antimicrobial peptides dermcidin and LL-37 . In this study we show that after secretion onto the skin surface, the CAMP gene product is processed by a serine protease-dependent mechanism into multiple novel antimicrobial peptides distinct from the cathelicidin LL-37 . These peptides show enhanced antimicrobial action, acquiring the ability to kill skin pathogens such as Staphylococcus aureus and Candida albicans . Furthermore, although LL-37 may influence the host inflammatory response by stimulating IL-8 release from keratinocytes, this activity is lost in subsequently processed peptides . Thus, a single gene product encoding an important defense molecule alters structure and function in the topical environment to shift the balance of activity toward direct inhibition of microbial colonization. J Immunol, 2004 Mar 1, 172(5), 3059 - 69 The contribution of the Toll-like/IL-1 receptor superfamily to innate and adaptive immunity to fungal pathogens in vivo; Bellocchio S et al.; In vitro studies have indicated the importance of Toll-like receptor (TLR) signaling in response to the fungal pathogens Candida albicans and Aspergillus fumigatus . However, the functional consequences of the complex interplay between fungal morphogenesis and TLR signaling in vivo remain largely undefined . In this study we evaluate the impact of the IL-1R/TLR/myeloid differentiation primary response gene 88 (MyD88)-dependent signaling pathway on the innate and adaptive Th immunities to C . albicans and A . fumigatus in vivo . It was found that 1) the MyD88-dependent pathway is required for resistance to both fungi; 2) the involvement of the MyD88 adapter may occur through signaling by distinct members of the IL-1R/TLR superfamily, including IL-1R, TLR2, TLR4, and TLR9, with the proportional role of the individual receptors varying depending on fungal species, fungal morphotypes, and route of infection; 3) individual TLRs and IL-1R activate specialized antifungal effector functions on neutrophils, which correlates with susceptibility to infection; and 4) MyD88-dependent signaling on dendritic cells is crucial for priming antifungal Th1 responses . Thus, the finding that the innate and adaptive immunities to C . albicans and A . fumigatus require the coordinated action of distinct members of the IL-1R/TLR superfamily acting through MyD88 makes TLR manipulation amenable to the induction of host resistance to fungi. Vet J, 2004 Mar, 167(2), 202 - 7 Intestinal candidiasis in a loggerhead sea turtle (Caretta caretta): an immunohistochemical study; Oros J et al.; Post mortem examination of a juvenile loggerhead sea turtle (Caretta caretta) stranded in the Canary Islands revealed a fishing-line in the small intestine . Histologically, severe necrotic enteritis, multiple haemorrhages, and marked oedema of the intestinal submucosa were observed . Yeast cells and fungal hyphae were seen in the lamina propria of the intestinal mucosa and in the connective tissue of the submucosa . Because fungal cultures were not taken at the time of necropsy, an immunohistochemical study was performed in order to identify the fungus involved . Specific monoclonal and heterologously absorbed polyclonal antibodies served as the primary reagents for identification of aspergillosis, candidiasis, fusariosis, geotricosis, scedosporiosis, and zygomycosis, using an indirect immunofluorescence staining technique . The fungal elements were strongly stained only by a polyclonal antibody against Candida albicans and a monoclonal antibody against C . albicans . There are no known previous reports of Candida sp . causing skin disease or systemic mycotic infection in sea turtles. Pol Merkuriusz Lek, 2003 Oct, 15(88), 388 - 90 {Multifocal fungal infections in patients after renal transplantation undergoing immunosuppression}; Kurnatowska I et al.; The aim of the study was to estimate the prevalence of fungi in monofocal and multifocal infections in renal transplant recipients and the characteristics of 60 species and intraspecies features of fungal strains isolated from the organ ontocenoses: oral cavity, rectum and genital organs in 32 patients undergoing permanent immunosuppression after renal transplantation . Fungal strains were recovered in 66 out of 96 samples (68.2% off all renal transplant recipients) . There were in ontocenoses: oral cavity (65.6%), rectum (37.5%) and genital organs (25.0%) . Monofocals mycosis were found only in 21.9% of the patients . Multifocal infections occurred in 68.2% and contained ontocenoses of oral cavity and rectum (34.4%) the most frequently . Trifocal infection occurred in 12.5% of all examined renal transplant recipients . Fungal strains identified using API 20 C and API 20 C AUX (bioMerieux) . The activity of 19 hydrolases was investigated using API ZYM . From among 41 strains of fungi the following were found: Candida albicans (31 strains), Candida glabrata (5), Candida guilliermondii (2), Candida krusei (2) and Saccharomyces cerevisiae (1) . The enzymograms were described for all strains and the highest activity was noted in case of: leucine arylamidase, acid phosphatase, esterase, naphtol-AS-BI-phosphohydrolase . The presence high mean of activity of this enzymes means high pathogenicity of C . albicans strains. J Arthroplasty, 2004 Feb, 19(2), 248 - 52 Candidal prosthetic hip infection in a patient with previous candidal septic arthritis; Lazzarini L et al.; We describe the case of a patient with chronic monocytic leukemia who underwent total hip arthroplasty (THA) for hip arthrosis . The patient has a history of Candida albicans arthritis of the same joint 5 months before THA surgery . Seven months after the prosthetic joint surgery, the patient developed a C albicans prosthetic infection that was successfully treated with amphotericin B and prosthesis removal . At surgery, the patient was believed cured of the candidal infection . Risk of infection after prosthetic joint surgery in patients with previous fungal joint infections has not been fully investigated . A lengthy infection-free follow-up period is probably necessary but may not be sufficient to prevent the occurrence of postoperative infections in these patients. Yeast, 2004 Feb, 21(3), 219 - 39 SRE1 and SRE2 are two specific steroid-responsive modules of Candida drug resistance gene 1 (CDR1) promoter; Karnani N et al.; CDR1 gene encoding an ATP-driven drug extrusion pump has been implicated in the development of azole-resistance in Candida albicans . Although the upregulation of CDR1 expression by various environmental factors has been documented, the molecular mechanism underlying such process is poorly understood . We have demonstrated earlier that the CDR1 promoter encompasses a large number of cis-regulatory elements, presumably mediating its response to various drugs . In this study we have identified a novel steroid responsive region (SRR) conferring beta-oestradiol and progesterone inducibility on the CDR1 promoter . The SRR is located -696 to -521 bp upstream of the transcription start site; it is modular in nature and can confer steroid responsiveness to a heterologous promoter (ADH1) linked to a GFP reporter gene . In vitro DNase I protection analyses of SRR revealed two progesterone responsive sequences (-628 to -594 and -683 to -648) and one beta-oestradiol responsive sequence (-628 to -577), which was further corroborated by the gel mobility shift assay . Deletion analyses within the SRR further delimited these steroid responsive sequences into two distinct elements, viz . SRE1 and SRE2 . While SRE1 (-677 to -648) responds only to progesterone, SRE2 (-628 to -598) responded to both progesterone and beta-oestradiol . Both SRE1 and SRE2 were specific for steroids, as they did not respond to other drugs, such as cycloheximide, miconazole and terbinafine . In silico comparison of the SRE1/2 with the promoter sequences of other MDR (CDR2 and PDR5) and non-MDR (HSP90) steroid-responsive genes revealed a similarity with respect to conservation of three 5 bp stretches (AAGAA, CCGAA and ATTGG) . Taken together, we have identified a novel steroid responsive cis-regulatory sequence in the CDR1 promoter, which presumably can be instrumental in understanding the steroid response cascade in Candida albicans . Biomaterials, 2004 Jul, 25(15), 3049 - 57 Conjugation of amino-containing drugs to polysaccharides by tosylation: amphotericin B-arabinogalactan conjugates; Ehrenfreund-Kleinman T et al.; The coupling of amphotericin-B (AmB), a water-insoluble antifungal and antileishmanial agent, to arabinogalactan (AG) via tosylate or mesylate derivatives was investigated as a method for the conjugation of amino-containing drugs to polysaccharides . In the first step, AG was reacted with tosyl- or mesyl-chloride at different ratios to obtain tosylate or mesylate AG derivatives . AmB was conjugated to AG derivatives in either aqueous or organic media via an amine bond . AG-AmB conjugates were soluble in water and exhibited improved stability in aqueous solutions as compared to the unbound drug . The conjugates showed comparable inhibitory concentration values against the pathogenic yeast Candida albicans, and against Leishmania major parasites . They were about 60 times less hemolytic against sheep erythrocytes than the free drug, and less toxic when injected i.v . to BALB/c mice. Pediatr Blood Cancer, 2004 Apr, 42(4), 332 - 7 Candida infections in children treated with conventional chemotherapy for solid tumors (transplant recipients excluded): The Institut Gustave Roussy Pediatrics Department experience; Ridola V et al.; INTRODUCTION: Advances in medical therapy have greatly improved the survival of children suffering from cancer . Although progress has been made in the eradication of malignant disease there is growing concern for the development of fungal infections in patients treated with chemotherapy . MATERIALS AND METHODS: We reviewed all episodes of pediatric candidemia that occurred between January 1988 and December 2000 . We analyzed the general characteristics of this population, risk factors, microbiology features, treatment, complications, and outcome . RESULTS: Seventeen cases of candidemia were observed during the 12 years of the study at an estimated incidence of 0.4% . Neutropenia occurred at the onset of infection in 13/17 (76.5%) children . A central venous device was present in all cases . Seventy-seven percent of the infections were caused by Candida albicans and in 85% of patients, yeasts had colonized the gastrointestinal tract . In 9/17 patients visceral dissemination was documented . Overall, in 77% of the episodes the outcome was favorable . CONCLUSIONS: Candidemia is a rare but severe complication in pediatric oncology . Even if the prognosis is better in children than in adults, Candida septicemia remains of great concern since a high percentage of these infections result in visceral dissemination and mortality is still elevated . Shanghai Kou Qiang Yi Xue, 2003 Aug, 12(4), 288 - 91 {The distribution of Candida spp . in the oral cavities of normal children}; Qi QG et al.; OBJECTIVE: To study the distribution of Candida spp . mainly Candida albicans in the oral cavities of health children . METHODS: Four groups children of different ages, A1: newborn babies, A2: 3.2 years old (average) children,A3, 7.2 years old,A4:12.7 years old and B control group 20.4 years old,mucosal swab sampling with centrification,CHROMagar Candida identified culture medium for culture and identification, and different methods for isolating Candida albicans for A3 group . RESULTS: The isolation rates of Candida spp were A1 7.5%,A2 70%,A3 56.36%,A4 49.12%,B 27.5%,the proportion of Candida albicans also diversely,and the method of PCR was more sensitive than the one of culture . CONCLUSION: Candida spp can be isolated from the normal oral cavities in different aged children, most of them were Candida albicans,both the isolation rates of Candida spp and the proportion of Candida albicans were different. Breast, 2002 Feb, 11(1), 88 - 90 Is lactation mastitis and shooting breast pain experienced by women during lactation caused by Candida albicans? Carmichael AR, Dixon JM. The evidence relating Candida albicans with deep breast pain suffered by some women during breast feeding is reviewed . There is little direct evidence to support the aetiological role of Candida albicans in this condition, and therefore the prescription of fluconazole (an antifungal drug) to lactating women with deep breast pain as suggested by a number of authors of studies on this subject cannot be justified unless as part of a clinical trial. Rev Assoc Med Bras, 2003 Oct-Dec, 49(4), 434 - 8 Epub 2004 Feb 04. {IgA, IgE and IgG subclasses to Candida albicans in serum and vaginal fluid from patients with vulvovaginal candidiasis}; de Carvalho RJ et al.; PURPOSE: To determine the levels of IgA, IgE, IgG and subclasses (IgG1, IgG4) antibodies specific to C . albicans in serum and vaginal washes from women with or without vulvovaginal candidiasis in order to evaluate the role of these antibodies in the immunopathogenesis of the disease . METHODS: Thirty women with clinical symptoms of vulvovaginal candidiasis (15 positive vaginal culture to C . albicans, 11 negative culture and 4 positive culture to non-C . albicans) and 12 asymptomatic control women were selected . Serum and vaginal wash samples were obtained for the detection of anti-C . albicans antibodies by ELISA . RESULTS: Symptomatic patients with positive culture showed significantly higher levels of specific IgA in vaginal washes and lower in serum than those with negative culture . Specific serum IgE levels were very low compared to vaginal IgE . High levels of total specific IgG were found in serum and vaginal washes in both groups, regardless the fungal presence or absence . Specific IgG1 and IgG4 levels were significantly higher only in vaginal washes from symptomatic patients with positive culture, with a slightly higher IgG1/IgG4 ratio, indicating that the IgG1 antibody response may be predominantly involved in the fungal clearance . CONCLUSION: Our results indicate a pronounced antibody response of IgA, IgG1 and IgG4 to C . albicans in vaginal washes in symptomatic patients with positive culture, suggesting a important role of these antibodies in the local immune response triggered by the presence of the fungus. J Antimicrob Chemother, 2004 Mar, 53(3), 526 - 9 Epub 2004 Feb 12. Susceptibility of clinical isolates of Candida lusitaniae to five systemic antifungal agents; Favel A et al.; OBJECTIVES: The aim of the present study was to expand the MIC database for Candida lusitaniae in order to further determine its antifungal susceptibility pattern . METHODS: The activities of amphotericin B, fluconazole, itraconazole, voriconazole and flucytosine were determined in vitro against 80 clinical isolates of C . lusitaniae . A set of 59 clinical isolates of Candida albicans and of 51 isolates of Candida glabrata was included to compare the susceptibilities to amphotericin B . The MICs were determined by Etest with RPMI 1640 agar, and with both this medium and antibiotic medium 3 (AM3) agar for testing of amphotericin B . RESULTS: All isolates were highly susceptible to fluconazole . The susceptibility to itraconazole was good; only 4% of isolates had dose-dependent susceptibility (MICs 0.25-0.5 mg/L) . Voriconazole was very active in vitro (100% of isolates were inhibited at < or =0.094 mg/L) . Flucytosine MICs ranged widely (0.004->32 mg/L) . The set included 19% of flucytosine-resistant isolates . For amphotericin B, 100% of isolates were inhibited at < or =0.75 mg/L (MIC(50) 0.047 mg/L; MIC(90) 0.19 mg/L) and at < or =4 mg/L (MIC(50) 0.25 mg/L; MIC(90) 0.75 mg/L) on RPMI and on AM3, respectively . A single isolate was categorized as resistant to amphotericin B (MIC 0.75 and 4 mg/L on RPMI and on AM3, respectively) . Amphotericin B thus appeared very active in vitro against C . lusitaniae . Whatever the test medium, the level of susceptibility of C . lusitaniae to amphotericin B did not differ much from those of C . albicans and C . glabrata . CONCLUSION: C . lusitaniae appears to be susceptible to amphotericin B, azole antifungal agents, and, to a lesser extent, flucytosine. Eukaryot Cell, 2004 Feb, 3(1), 190 - 9 Candida albicans lacking the gene encoding the regulatory subunit of protein kinase A displays a defect in hyphal formation and an altered localization of the catalytic subunit; Cassola A et al.; The fungal pathogen Candida albicans switches from a yeast-like to a filamentous mode of growth in response to a variety of environmental conditions . We examined the morphogenetic behavior of C . albicans yeast cells lacking the BCY1 gene, which encodes the regulatory subunit of protein kinase A . We cloned the BCY1 gene and generated a bcy1 tpk2 double mutant strain because a homozygous bcy1 mutant in a wild-type genetic background could not be obtained . In the bcy1 tpk2 mutant, protein kinase A activity (due to the presence of the TPK1 gene) was cyclic AMP independent, indicating that the cells harbored an unregulated phosphotransferase activity . This mutant has constitutive protein kinase A activity and displayed a defective germinative phenotype in N-acetylglucosamine and in serum-containing medium . The subcellular localization of a Tpk1-green fluorescent protein (GFP) fusion protein was examined in wild-type, tpk2 null, and bcy1 tpk2 double mutant strains . The fusion protein was observed to be predominantly nuclear in wild-type and tpk2 strains . This was not the case in the bcy1 tpk2 double mutant, where it appeared dispersed throughout the cell . Coimmunoprecipitation of Bcy1p with the Tpk1-GFP fusion protein demonstrated the interaction of these proteins inside the cell . These results suggest that one of the roles of Bcy1p is to tether the protein kinase A catalytic subunit to the nucleus. Eukaryot Cell, 2004 Feb, 3(1), 135 - 43 Candida albicans Csy1p is a nutrient sensor important for activation of amino acid uptake and hyphal morphogenesis; Brega E et al.; Candida albicans is an important human pathogen that displays a remarkable ability to detect changes in its environment and to respond appropriately by changing its cell morphology and physiology . Serum- and amino acid-based media are known to induce filamentous growth in this organism . However, the mechanism by which amino acids induce filamentation is not yet known . Here, we describe the identification and characterization of the primary amino acid sensor of C . albicans, Csy1 . We show that Csy1p plays an important role in amino acid sensing and filamentation . Loss of Csy1p results in a lack of amino acid-mediated activation of amino acid transport and a lack of induction of transcription of specific amino acid permease genes . Furthermore, a csy1Delta/csy1Delta strain, lacking Csy1p, is defective in filamentation and displays altered colony morphology in serum- and amino acid-based media . These data provide the first evidence that C . albicans utilizes the amino acid sensor Csy1p to probe its environment, coordinate its nutritional requirements, and determine its morphological state. Farmaco, 2004 Feb, 59(2), 163 - 7 Efficacy of a new ketoconazole bioadhesive vaginal tablet on Candida albicans; Karasulu HY et al.; To develop more effective treatment for vaginal candidasis, ketoconazole (KTZ) was formulated in bioadhesive tablet formulations that increase the time of contact of drug with the vaginal mucosa . The bioadhesive vaginal tablets delivery of KTZ was prepared by direct compression of sodium carboxymethyl cellulose or polyvinylpyrrolidone or hydroxypropylmethyl cellulose (HPMC-E(50)) . Dissolution studies of bioadhesive tablets and commercial ovules were carried out with a new basket method (horizontal rotating basket) . In vitro, a good sustained release action was obtained with bioadhesive tablets containing 1:1 and 1:2 drug/polymer ratio using HPMC-E(50) . These bioadhesive tablets containing 400 mg of KTZ showed a zero-order drug release kinetic . KTZ solutions at increasing concentrations (0.16, 0.33, 0.5 and 0.66 mg/ml) were prepared for microbiological trials . These concentrations correspond to 25%, 50%, 75% and 100% of KTZ released from bioadhesive tablets, respectively . Yeast mixture was mixed with each concentration of KTZ at ratio of 1:10 . One hundred microliters of this mixture was transferred in 900 microl liquid Sabouraud medium after a certain time interval for each concentration of KTZ and incubation at 37 degrees C for 24 h . Then this culture streaked onto Sabouraud-dextrose-agar plates, which were incubated at 37 degrees C for 48 h . The 0.16 and 0.33 mg/ml concentrations of KTZ showed fungistatic effect in 120 min . The 0.5 mg/ml concentration of KTZ was fungistatic in 90 and 120 min; and the 0.66 mg/ml concentration of the drug was fungistatic in 120 min as well as in 180 min . It was found that, in vitro antifungal activity of KTZ was dependent on its concentration and contact time with yeast cells . These results indicated that a new bioadhesive vaginal tablet formulations might be further developed for safe convenient and effective treatment of vaginal candidasis. Oral Microbiol Immunol, 2004 Apr, 19(2), 111 - 7 Expression and regulation of novel human beta-defensins in gingival keratinocytes; Premratanachai P et al.; This study evaluated the expression and regulation of beta-defensins DEFB-104 and the recently identified DEFB-105-14 in gingival keratinocytes . Keratinocytes from healthy subjects were exposed to cytokines, Escherichia coli lipopolysaccharide or Candida species . Total RNA was extracted and defensin expression analyzed by reverse transcription-polymerase chain reaction . Three patterns of expression were seen: no expression, constitutive expression and inducible expression . Constitutive mRNA expression was evident for DEFB-104, 107, 109, 111, and 112 . DEFB-108 and 114 were induced by interleukin (IL)-1beta and Candida species . For DEFB-108 expression, synergism was observed when IL-1beta was combined with tumor necrosis factor-alpha or interferon-gamma . Downregulation of DEFB-109 occurred following treatment with Candida albicans . These findings suggest a role for multiple beta-defensins in response to oral infection . Further investigation is needed to better understand their function, both in terms of antimicrobial activities and contributions to innate and acquired immunity. J Cell Biol, 2004 Feb 16, 164(4), 581 - 91 Epub 2004 Feb 09. In Candida albicans, the Nim1 kinases Gin4 and Hsl1 negatively regulate pseudohypha formation and Gin4 also controls septin organization; Wightman R et al.; In the development of hyphal germ tubes of Candida albicans, a band of septin forms at the base of the germ tube (basal septin band) . Later, a septin ring forms, which organizes the first septum within the germ tube (septin ring) . We have investigated the role of the Nim1 kinases, Gin4 and Hsl1, in the formation of these septin structures . We show that during germ tube formation, Gin4 is required for the organization of the septin ring but not the basal septin band . Hsl1 is not required for the formation of either septin rings or basal bands . Unexpectedly, we found that both gin4Delta and hsl1Delta mutants form pseudohyphae constitutively, in a fashion that in the case of gin4Delta, is partly independent of Swe1 . Gin4-depleted pseudohyphae are unable to form hyphae when challenged with serum, but this can be overcome by ectopic expression of Gin4 from the MET3 promoter . Thus, Gin4 may regulate the developmental switch from pseudohyphae to hyphae . Copyright The Rockefeller University Press Neth J Med, 2003 Nov, 61(11), 365 - 9 Candida-specific interferon-gamma deficiency and toll-like receptor polymorphisms in patients with chronic mucocutaneous candidiasis; van der Graaf CA et al.; Chronic mucocutaneous candidiasis (CMC) is a group of disorders, characterised by persistent mucocutaneous infections with Candida species . The underlying defect of CMC has not been elucidated, but a defective cytokine response may be involved . Therefore, we investigated whether an imbalance between IFNgamma and IL-10 may play a role in this disorder . We assessed the cytokine production in whole-blood cultures from CMC patients using Candida albicans, lipopolysaccharide and phytohaemagglutinin as stimuli . As the Toll-like receptors are important pattern recognition receptors for Candida species, we also investigated Toll-like receptor polymorphisms in these patients . Patients with CMC had a significantly decreased IFNgamma production when whole blood was stimulated with C . albicans (232 +/- 120 vs 2279 +/- 609 pg/ml, p<0.02) . When stimulated with phytohaemagglutinin, the differences were not significant (3549 +/- 1320 vs 7631 +/- 1790 pg/ml) . The Candida-stimulated production of IL-10 tended to be higher in CMC patients, whereas TNF and IL-1beta production were similar in patients and controls . Stimulation with LPS showed no differences in cytokine production between patients and controls . Two out of seven patients had the TLR4 Asp299Gly polymorphism and none had the TLR2 Arg677Trp polymorphism . These data support the hypothesis that deficient IFNgamma production is involved in the pathogenesis of CMC, whereas a role for genetic polymorphisms of Toll-like receptor 2 and 4 is not obvious in these patients. Jpn J Ophthalmol, 2004 Jan-Feb, 48(1), 30 - 3 Bilateral Candida albicans endophthalmitis associated with an infected deep venous thrombus; Arroyo JG et al.; BACKGROUND: To describe the clinical and histopathologic findings in a patient with bilateral Candida albicans endophthalmitis due to an infected deep venous thrombus . CASE: A 43-year-old patient with bilateral Candida albicans endophthalmitis due to an infected central venous thrombus . The patient's ophthalmology and hospital charts were reviewed . Histopathologic sections of the Candida albicans retinal granuloma were examined under light microscopy . OBSERVATIONS: Our patient had multiple blood cultures positive for Candida albicans owing to an infected subclavicular venous thrombosis . Bilateral Candida albicans endophthalmitis was diagnosed . Bilateral vitrectomy and membrane peeling for macular traction retinal detachments were performed . In the right eye, a large retinal granuloma was excised during surgery to adequately relieve traction on the macula . Vision improved in both eyes after surgery . Histopathologic findings revealed branching hyphae surrounded by giant cells, endothelial cell-lined vascular channels, and inflammatory cells . CONCLUSIONS: This is the first report of an infected deep venous thrombosis causing bilateral endogenous endophthalmitis . Appropriate management of these patients requires clear differentiation between endogenous chorioretinitis and endophthalmitis . Patients with documented fungemia should have a dilated fundus examination on a regular basis until complete clearance of the infection. Microbiology, 2004 Feb, 150(Pt 2), 285 - 92 Differential surface localization and temperature-dependent expression of the Candida albicans CSH1 protein; Singleton DR et al.; Cell-surface hydrophobicity (CSH) in Candida albicans contributes to virulence and can be conveniently regulated in planktonic cultures by altering growth temperature . The CSH1 gene is the first candidate gene that has been demonstrated to play a role in affecting the CSH phenotype . However, the primary amino acid sequence of the CSH1 gene product suggests that the protein should be restricted to the cytoplasm . A majority of the protein appears to demonstrate that localization . Cell-surface biotinylation and limited glucanase digestion were used to determine and estimate the relative amount of Csh1p in the extracellular compartment in comparison to the cytoplasmic pool . Additionally, Western and Northern blotting were used to assess expression of the CSH1 gene under different growth conditions . Compared with cells grown at 23 degrees C, the total cellular levels of Csh1p are significantly greater at elevated growth temperatures . Detection of Csh1p on the cell surface correlates with the level of overall protein expression . The temperature-dependent regulation and surface presentation of Csh1p suggests a mechanism for regulating the CSH phenotype. Microbiology, 2004 Feb, 150(Pt 2), 277 - 84 Accessibility of the peptide backbone of protein ligands is a key specificity determinant in Candida albicans SRS adherence; Gaur NK et al.; Candida albicans displays a high degree of specificity in selecting and adhering to targets in vivo . The features of target recognition are poorly understood and likely to involve more than the mere chemical composition of the ligand . Using an adherence assay in which protein and peptide ligands are covalently coupled to magnetic beads, the authors have previously described a new adherence mechanism in C . albicans, henceforth referred to as SRS (stable, reversible, specific) adherence . It was previously demonstrated that C . albicans and Saccharomyces cerevisiae expressing agglutinin-like sequence 5 protein (Als5p, previously referred to as Ala1p or Ala1/Als5p) adhere to peptides containing patches of threonine, serine and alanine residues when these are located in the free end of immobilized peptides . The interaction with protein ligands in SRS adherence predominantly involves the formation of hydrogen bonds . Accordingly, this interaction may occur (1) to the peptide backbone of the protein ligand or (2) to the amino acid side chain with an appropriate functional group . Evidence is provided that the primary interaction occurs with the peptide backbone and the secondary interaction occurs with the side chain . The primary interaction with the peptide backbone is sufficient for adherence to occur, whereas the secondary interaction with a side chain possessing an appropriate functional group stabilizes the interaction . In agreement with these results, it is also demonstrated that proteins lacking secondary and tertiary structure, wherein the peptide backbone is sterically accessible, interact with C . albicans and S . cerevisiae expressing Als5p . C . albicans Als proteins are resistant to denaturation by harsh conditions that kill the yeast cells . The proposed interactions in SRS adherence have striking similarities with those of the molecular chaperone Hsp70, which specifically binds to non-native proteins and resists denaturation. J Clin Microbiol, 2004 Feb, 42(2), 899 - 902 Comparison of two microdilution methods for testing susceptibility of Candida spp . to voriconazole; Linares MJ et al.; The growing number of fungal infections, coupled with emerging resistance to classical antifungal agents, has led to the development of new agents, among them voriconazole . Susceptibility to voriconazole was tested by using two microdilution techniques: the reference method described in National Committee for Clinical Laboratory Standards document M27-A2 and a colorimetric method, Sensititre YeastOne . A total of 272 Candida isolates (132 of Candida albicans, 62 of C . parapsilosis, 33 of Candida glabrata, 21 of C . krusei, 15 of C . tropicalis, and 9 of C . lusitaniae) and two control strains (C . parapsilosis ATCC 22019 and C . krusei ATCC 6258) were tested . There was a high rate of agreement between the two methods used (97 to 100%). J Clin Microbiol, 2004 Feb, 42(2), 773 - 7 Comparison of Mycosis IC/F and plus Aerobic/F media for diagnosis of fungemia by the bactec 9240 system; Meyer MH et al.; Fungemia is associated with a high mortality rate . We compared the performance of the Mycosis IC/F selective fungal medium and the Plus Aerobic/F standard bacteriological medium for the diagnosis of fungemia on the Bactec 9240 automatic system . We retrospectively analyzed 550 blood culture pairs composed of one Mycosis IC/F vial and one Plus Aerobic/F vial, drawn in 187 patients with fungemia . The positivity rate by vial was significantly higher on Mycosis IC/F medium than on Plus Aerobic/F medium (88.0% versus 74.9%, P < 0.0001) . The positivity rate for fungus detection on Plus Aerobic/F medium fell to 26.9% when bacteria were present in the same vial . The positivity rate by patient was also significantly higher on Mycosis IC/F medium than on Plus Aerobic/F medium (92.5% versus 75.9%, P < 0.0001) . A marked superiority of Mycosis IC/F medium was demonstrated for diagnosis of Candida glabrata fungemia (31 of 31, 100%, versus 18 of 31, 58.1%, P < 0.0001) . The mean detection time was significantly shorter on Mycosis IC/F medium than on Plus Aerobic/F medium (28.9 +/- 22.2 h versus 36.5 +/- 24.6 h, P < 0.0001) . The mean time saving was 8.8 h for Candida albicans and 43.7 h for C . glabrata . Mycosis IC/F medium enabled more sensitive and earlier diagnosis, particularly for the two strains most frequently responsible for fungemia, C . albicans and C . glabrata, and also in the event of the concomitant presence of both yeasts and bacteria . In patients with risk factors, it would thus appear to be sensible to draw a Mycosis IC/F vial in addition to the standard bacteriological vials. Planta Med, 2004 Jan, 70(1), 90 - 2 Antifungal steroid saponins from Dioscorea cayenensis; Sautour M et al.; From the rhizomes of Dioscorea cayenensis Lam.-Holl (Dioscoreaceae), the new 26- O- beta- D-glucopyranosyl-22-methoxy-3 beta,26-dihydroxy-25( R)-furost-5-en-3- O- alpha- L-rhamnopyranosyl-(1-->4)- alpha- L-rhamnopyranosyl-(1-->4)-{ alpha- L-rhamnopyranosyl-(1-->2)}- beta- D-glucopyranoside ( 1) was isolated together with the known dioscin ( 2) and diosgenin 3- O- alpha- L-rhamnopyranosyl-(1-->4)- alpha- L-rhamnopyranosyl-(1-->4)-{ alpha- L-rhamnopyranosyl-(1-->2)}- beta- D-glucopyranoside ( 3) . Their structures were established on the basis of spectral data . Compound 2 exhibited antifungal activity against the human pathogenic yeasts Candida albicans, C . glabrata and C . tropicalis (MICs of 12.5, 12.5 and 25 micro g/mL, respectively) whereas 3 showed weak activity and 1 was inactive. Blood, 2004 Jun 1, 103(11), 4322 - 9 Epub 2004 Feb 05. T lymphocytes of recipient origin may contribute to the recovery of specific immune response toward viruses and fungi in children undergoing cord blood transplantation; Montagna D et al.; Patients undergoing allogeneic cord blood transplantation (CBT) benefit from a low risk of graft-versus-host disease (GVHD), but there are still concerns that they be able to recover an effective immune capacity early after transplantation . We investigated the ability to develop in vitro T-lymphocyte-mediated immune response toward human cytomegalovirus and Candida albicans antigens, early and late after transplantation, in children given cord blood transplants from either an HLA-identical sibling or an unrelated donor . Proliferative capacity and frequency of antigen-specific T cells were evaluated; antigen-specific CD4(+) T-cell clones were also generated and characterized for T-cell receptor repertoire diversity, cytokine phenotype, and their origin (either from donor or patient) . We found that the majority of recipients can develop a specific response to viral or fungal antigens already early after transplantation . Antigen-specific T-cell clones of both donor and recipient origin contributed to the reconstitution of immune response . Antigen-specific T lymphocytes of recipient origin were detected in patients receiving a transplant from a relative, after a chemotherapy-based conditioning regimen, and who did not have GVHD . Our results document, at a clonal level, that after CBT recovery of either polyclonal or pauciclonal T-cell response toward widespread pathogens is prompt, with some patients benefiting from a contribution of recipient-derived cells. Mem Inst Oswaldo Cruz, 2003 Oct, 98(7), 959 - 61 Epub 2004 Jan 07. Antimicrobial activity in vitro of plumbagin isolated from Plumbago species; de Paiva SR et al.; Plumbagin is a naturally occurring naphthoquinone isolated from roots of Plumbago scandens . The plant was collected at the Campus of Fundacao Oswaldo Cruz, Rio de Janeiro, Brazil . P . scandens is used as a traditional medicine for the treatment of several diseases . The antimicrobial activity of plumbagin was evaluated using the macrodilution method . The compound exhibited relatively specific activity against bacteria and yeast . The minimum inhibitory concentration test showed the growth inhibiton of Staphylococcus aureus at a concentration of 1.56 g/ml and of Candida albicans at a concentration of 0.78 g/ml . These results suggest the naphthoquinone plumbagin as a promising antimicrobial agent. Enferm Infecc Microbiol Clin, 2004 Feb, 22(2), 83 - 8 {Evaluation of a new commercial test (Candida albicans IFA IgG) for the serodiagnosis of invasive candidiasis}; Moragues MD et al.; INTRODUCTION: Two tests for the detection of antibodies to Candida albicans germ tubes in patients with invasive candidiasis were compared: a new commercially available test (Candida albicans IFA IgG) and the indirect immunofluorescence test generally used for this purpose . METHODS: With the use of two indirect immunofluorescence tests, retrospective study was done on 172 sera from 51 patients classified into two groups: Group I included 123 serum samples from 32 patients with invasive candidiasis, and Group II, the control, included 49 serum samples from 19 patients with no evidence of Candida infection . RESULTS: In Group I, 84% of patients presented anti-germ tube antibody titers >or= 1:160 by the Candida albicans IFA IgG test and 78.1% of patients were positive by the generally used test . There was a high correlation between the two tests (R2 =0.9512 by patients; R2 = 0.8986 by sera) . When a titer value of >or= 1:160 was used as cutoff, the Candida albicans IFA IgG test showed a sensitivity of 84.4% and a specificity of 94.7%, whereas the traditional test showed a sensitivity of 78.1% and a specificity of 100% . CONCLUSIONS: The commercially available Candida albicans IFA IgG test is similar to the test generally used for the detection of antibodies to C . albicans germ tubes and provides faster and easier diagnosis of invasive candidiasis in the clinical microbiology laboratory. Mol Microbiol, 2004 Jan, 51(2), 551 - 65 Chromosome 1 trisomy compromises the virulence of Candida albicans; Chen X et al.; Although increases in chromosome copy number typically have devastating developmental consequences in mammals, fungal cells such as Saccharomyces cerevisiae seem to tolerate trisomies without obvious impairment of growth . Here, we demonstrate that two commonly used laboratory strains of the yeast Candida albicans, CAI-4 and SGY-243, can carry three copies of chromosome 1 . Although the trisomic strains grow well in the laboratory, Ura+ derivatives of CAI-4, carrying three copies of chromosome 1, are avirulent in the intravenously inoculated mouse model, unlike closely related strains carrying two copies of chromosome 1 . Furthermore, changes in chromosome copy number occur during growth in an animal host and during growth in the presence of growth-inhibiting drugs . These results suggest that chromosome copy number variation provides a mechanism for genetic variation in this asexual organism. Mol Microbiol, 2004 Jan, 51(2), 371 - 84 An ER packaging chaperone determines the amino acid uptake capacity and virulence of Candida albicans; Martinez P et al.; The Candida albicans CSH3 gene encodes a functional and structural homologue of Shr3p, a yeast protein that is specifically required for proper uptake and sensing of extracellular amino acids in Saccharomyces cerevisiae . A Candida csh3delta/csh3delta null mutant has a reduced capacity to take up amino acids, and is unable to switch morphologies on solid and in liquid media in response to inducing amino acids . CSH3/csh3delta heterozygous strains display normal amino acid induced morphological switching . However, although heterozygous cells apparently sense and properly react to amino acid induced signals they cannot take up amino acids at wild-type rates . Strikingly, both CSH3/csh3delta heterozygous and csh3delta/csh3delta homozygous strains are unable to efficiently mount virulent infections in a mouse model . The haploinsufficiency phenotypes indicate that both CSH3 alleles contribute to maintain high-capacity amino acid uptake in wild-type strains . These results strongly suggest that C . albicans cells use amino acids, presumably as nitrogen sources, during growth in mammalian hosts. Proteomics, 2004 Feb, 4(2), 374 - 82 Two-dimensional reference map of Candida albicans hyphal forms; Hernandez R et al.; We have set up an in vitro model of culture of Candida albicans fungal cells that mimics the physiological conditions found in human beings in order to carry out studies of host-pathogen interaction . Under these conditions, C . albicans undergoes dimorphic transition (yeast-hyphae); this morphological change has been proposed as a virulence factor for this fungus . C . albicans cytoplasmic extract from hyphae cells was analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and a reference map was obtained . Protein identification was carried out by peptide mass fingerprinting or sequence tagging using a matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) or a MALDI-TOF/TOF mass spectrometer . A total of 106 spots, excised from 2-D gels, were analyzed . This resulted in the identification of 43 proteins involved in metabolism, 13 involved in transcription, protein synthesis, and fate, 8 involved in cell rescue, virulence, and defense, and 2 proteins of unknown function . This reference map is an important tool for future studies of protein differential expression after host-pathogen interaction. Bioorg Med Chem, 2004 Feb 15, 12(4), 779 - 87 Design of tetrapeptide ligands as inhibitors of the Src SH2 domain; Nam NH et al.; Src homology-2 (SH2) domains are noncatalytic motifs containing approximately 100 amino acid residues that are involved in intracellular signal transduction . The phosphotyrosine-containing tetrapeptide pTyr-Glu-Glu-Ile (pYEEI) binds to Src SH2 domain with high affinity (K(d)=100 nM) . The development of five classes of tetrapeptides as inhibitors for the Src SH2 domain is described . Peptides were prepared via solid-phase peptide synthesis and tested for affinity to Src SH2 domain using a fluorescence polarization based assay . All of the N-terminal substituted pYEEI derivatives (class II) presented binding affinity (IC(50)=of 2.7-8.6 microM) comparable to pYEEI (IC(50)=6.5 microM) in this assay . C-Terminal substituted pYEEI derivatives (class III) showed a lower binding affinity with IC(50) values of 34-41 microM . Amino-substituted phenylalanine derivatives (class IV) showed weak binding affinities (IC(50)=16-153 microM) . Other substitutions on phenyl ring (class I) or the replacement of the phenyl ring with other cyclic groups (class V) dramatically decreased the binding of tetrapeptides to Src SH2 (IC(50)>100 microM) . The ability of pYEEI and several of the tetrapeptides to inhibit the growth of cancer cells were assessed in a cell-based proliferation assay in human embryonic kidney (HEK) 293 tumor cells . The binding affinity of several of tested compounds against Src SH2 domain correlates with antiproliferative activity in 293T cells . None of the compounds showed any significant antifungal activity against Candida albicans ATCC 14053 at the maximum tested concentration of 10 microM . Overall, these results provided the structure-activity relationships for some FEEI and YEEI derivatives designed as Src SH2 domain inhibitors. Skin Pharmacol Physiol, 2004 Jan-Feb, 17(1), 31 - 6 Toxicity and antimicrobial activity of a hydrocolloid dressing containing silver particles in an ex vivo model of cutaneous infection; Schaller M et al.; In the present study we examined the effects of two hydrocolloid wound dressings (conventional silver-free Comfeel, silver-incorporating Contreet-H) on uninfected and Candida albicans- or methicillin-resistant Staphylococcus aureus-infected reconstituted human epithelium (RHE) . The morphological alterations of the keratinocytes caused by infection and by treatment were analysed with light and electron microscopy . As a measure of epithelial cell damage the release of lactate dehydrogenase from epithelial cells into the surrounding medium was monitored . Appl