Yeasts

Arch Biochem Biophys, 1997 Aug 15, 344(2), 295 - 300
Characterization of a pollination-related cDNA from Phalaenopsis encoding a protein which is homologous to human peroxisomal acyl-CoA oxidase; Do YY et al.; The first putative plant acyl-CoA oxidase cDNA has been isolated from a Phalaenopsis cDNA library constructed by poly(A)+ RNA extracted from petals 1 day after pollination . This cDNA, pOACO31, contains a 2100-bp open reading frame which encodes a polypeptide named PACO1 of 699 amino acids . The predicted isoelectric point of PACO1 is 8.74 and the molecular weight is 78,032 Da, similar to that of a monomer of predicted plant acyl-CoA oxidase . Southern blot analysis indicated that this gene occurs in one copy or a low number of copies per haploid genome . When compared with sequences in databases, PACO1 revealed significant similarity only to peroxisomal acyl-CoA oxidase particularly within 13 conserved regions and a putative FMN-binding site.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1835 - 6
In vitro antifungal activity of pneumocandin L-743,872 against a variety of clinically important molds; Del Poeta M et al.; The in vitro activity of the new antifungal drug pneumocandin L-743,872 against 55 isolates of clinically important molds was examined by an adapted macrobroth dilution method for yeasts . Pneumocandin L-743,872 exhibited in vitro antifungal activity against Alternaria sp., Aspergillus flavus, Aspergillus fumigatus, Curvularia lunata, Exophiala jeanselmei, Fonsecaea pedrosoi, Paecilomyces variotii, and Scedosporium apiospermum . The drug appeared to lack significant in vitro inhibitory activity against Fusarium oxysporum, Fusarium solani, Rhizopus arrhizus, Paecilomyces lilacinus, and Scedosporium prolificans.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1832 - 4
In vitro evaluation of voriconazole against some clinically important fungi; McGinnis MR et al.; Voriconazole was compared to amphotericin B, fluconazole, and itraconazole by using an in vitro macrobroth dilution test based upon current National Committee for Clinical Laboratory Standards tentative standards against the dimorphic fungi and several opportunistic molds and yeasts . In all instances, the voriconazole MICs were lower than those of fluconazole . In most instances, the MICs were lower than the recorded MICs of amphotericin B and itraconazole.

J Clin Microbiol, 1997 Aug, 35(8), 2031 - 9
Identification of Candida species by randomly amplified polymorphic DNA fingerprinting of colony lysates; Steffan P et al.; We have characterized a method that produces simple yet diagnostic fingerprints that are unique to isolates of Candida species . DNA from individual colonies can be amplified from crude single-colony lysates . Randomly amplified polymorphic DNA (RAPD) fingerprints generated from a single primer correctly identified the species of most (>98%) of the isolates identified with CHROMagar Candida plates as non-Candida albicans Candida species . RAPD fingerprints were much more informative than the plates, since they distinguished between all tested species and required less time . Most (91%) of these identifications agreed with those assigned by API 20C tests . In almost every incident of species identity mismatch, electrophoretic karyotyping showed that the RAPD fingerprint was correct . This underscores the improved objectivity and reliability of this method over those of conventional diagnostic tools . The identities of approximately 30% of C . albicans isolates identified in clinical laboratories by positive germ tube tests are not verified by either testing on CHROMagar Candida plates or RAPD fingerprinting . Data suggest that clinical isolates conventionally identified as C . albicans in clinical settings are heterogeneous, consisting of both misidentified and atypical yeasts . RAPD fingerprints obtained from primary culture plate colonies allows for rapid, highly accurate determinations of Candida species, hence permitting earlier selection of appropriate antifungal agents in the clinical setting.

Can J Microbiol, 1997 Jul, 43(7), 649 - 57
Murine macrophage elastolytic activity induced by Aspergillus fumigatusstrains in vitro: evidence of the expression of two macrophage-induced protease genes; Rodriguez E et al.; The interaction between Aspergillus fumigatus conidia and murine macrophages of various origins was investigated . Cocultures were carried out between A . fumigatus strains and freshly isolated murine pulmonary alveolar macrophages or two murine macrophage cell-lines: murine alveolar cell-line MALU and murine astrocytoma cell-line J774 . By measuring the variation of elastolytic activity in the coculture supernatants with two elastin substrates, we demonstrated that either viable or fixed A . fumigatus or C . albicans yeasts or nonspecific particles induced significant macrophage elastolytic activity . The effect of A . fumigatus supernatant or the purified A . fumigatus galactomannan suggested also the possible involvement of this polysaccharide in macrophage-protease gene expression, release, and activity in invasive aspergillosis . The effect of inhibitory compounds demonstrated the potential implication of a macrophagic metalloprotease and a macrophagic cysteine protease . RNA analysis allowed us to demonstrate the induction of expression of two macrophagic protease genes in stimulated macrophages . Two distinctive mechanisms appeared to be implicated in macrophage protease induction: nonspecific phagocytosis in the earliest times of the coculture and (or) specific galactomannan recognition after its gradual release by the mycelium.

Infect Immun, 1997 Jul, 65(7), 2564 - 9
Intrapulmonary response to Histoplasma capsulatum in gamma interferon knockout mice; Allendoerfer R et al.; We examined the immunobiological responses to Histoplasma capsulatum in lungs of gamma interferon (IFN-gamma) knockout mice (GKO mice) . Naive GKO mice succumbed by day 9 to intranasal challenge with 2.5 x 10(6) yeasts, whereas all wild-type (WT) mice survived for 45 days . Compared to lungs of WT mice, the lungs of acutely infected GKO mice exhibited dramatically elevated numbers of CFU in lungs and significantly higher levels of tumor necrosis factor alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF) but not interleukin-12 (IL-12) or IL-4 . To determine if IFN-gamma is necessary in reexposure histoplasmosis, GKO and WT mice were inoculated with 10(4) yeasts intranasally and given amphotericin B for 3 weeks . Six weeks later, mice were rechallenged with 2.5 x 10(6) yeasts . All GKO mice died by day 6, whereas all WT mice survived for 45 days . Lungs of GKO mice contained substantially elevated numbers of CFU and higher TNF-alpha and GM-CSF levels but not IL-12 or IL-4 . Thus, IFN-gamma is requisite for control of pulmonary histoplasmosis in naive and reexposed mice.

Cell, 1997 Jun 27, 89(7), 1155 - 64
Coassembly of TRP and TRPL produces a distinct store-operated conductance; Xu XZ et al.; The Drosophila retinal-specific protein, TRP (transient receptor potential), is the founding member of a family of store-operated channels (SOCs) conserved from C . elegans to humans . In vitro studies indicate that TRP is a SOC, but that the related retinal protein, TRPL, is constitutively active . In the current work, we report that coexpression of TRP and TRPL leads to a store-operated, outwardly rectifying current distinct from that owing to either TRP or TRPL alone . TRP and TRPL interact directly, indicating that the TRP-TRPL-dependent current is mediated by heteromultimeric association between the two subunits . We propose that the light-activated current in photoreceptor cells is produced by a combination of TRP homo- and TRP-TRPL heteromultimers.

Cell, 1997 Jun 27, 89(7), 1067 - 76
Daxx, a novel Fas-binding protein that activates JNK and apoptosis; Yang X et al.; The Fas cell surface receptor induces apoptosis upon receptor oligomerization . We have identified a novel signaling protein, termed Daxx, that binds specifically to the Fas death domain . Overexpression of Daxx enhances Fas-mediated apoptosis and activates the Jun N-terminal kinase (JNK) pathway . A C-terminal portion of Daxx interacts with the Fas death domain, while a different region activates both JNK and apoptosis . The Fas-binding domain of Daxx is a dominant-negative inhibitor of both Fas-induced apoptosis and JNK activation, while the FADD death domain partially inhibits death but not JNK activation . The Daxx apoptotic pathway is sensitive to both Bcl-2 and dominant-negative JNK pathway components and acts cooperatively with the FADD pathway . Thus, Daxx and FADD define two distinct apoptotic pathways downstream of Fas.

J Biol Chem, 1997 Jun 20, 272(25), 15834 - 40
Expression cloning of PIG-L, a candidate N-acetylglucosaminyl-phosphatidylinositol deacetylase; Nakamura N et al.; Many eukaryotic cell surface proteins are bound to the cell membrane by a glycosylphosphatidylinositol (GPI) anchor . Several genes involved in GPI anchor biosynthesis have been cloned using complementation of mutant mammalian cell lines and yeasts that are defective in its biosynthesis pathway . However, the gene involved in the second step of this pathway, in which N-acetylglucosaminyl-phosphatidylinositol (GlcNAc-PI) is N-deacetylated to form glucosaminyl (GlcN)-PI, has not been cloned . In this study, we established a GPI anchor-deficient mutant of Chinese hamster ovary (CHO) cells defective in the second step . Complementation analysis with the known GPI anchor mutant cells demonstrated that it belonged to the same complementation group as the CHO cell mutant G9PLAP.85 . Using the new mutant, we cloned a rat gene termed PIG-L (for phosphatidylinositol glycan class L) that is involved in this step . PIG-L encodes a 252-amino acid, endoplasmic reticulum membrane protein, most of which is in the cytoplasmic side . This orientation of PIG-L protein is consistent with the notion that the second step of GPI anchor biosynthesis occurs on the cytoplasmic side of the endoplasmic reticulum.

Cell, 1997 Jun 13, 89(6), 859 - 66
The eucaryal tRNA splicing endonuclease recognizes a tripartite set of RNA elements; Di Nicola Negri E et al.; The tRNA splicing endonuclease cleaves intron-containing tRNA precursors on both sides of the intron . The prevailing belief has been that the enzyme binds only to the mature domain through the invariant bases . We show instead that, for recognition, the endonuclease utilizes distinct sets of structural elements, several of which are within the intron . One subset of recognition elements, localized in the mature domain, is needed for recognition of both cleavage sites, while two other subsets, localized at the exon-intron boundaries, are used for recognition of either one or the other cleavage site . The two cleavage sites are essentially independent: neither is required by the other for cleavage to take place . These results support a two-active-site model for the eucaryal endonuclease.

J Am Osteopath Assoc, 1997 Jun, 97(6), 339 - 46
Advances in the treatment of superficial fungal infections: focus on onychomycosis and dry tinea pedis; Del Rosso JQ; Onychomycosis is one of the most stubborn superficial mycoses . With few exceptions, oral antifungal therapy is needed to achieve resolution . Before oral itraconazole, fluconazole, and terbinafine hydrochloride became available, physicians had to rely on prolonged therapy with griseofulvin or oral ketoconazole . Of the newer oral agents, itraconazole appears to have the broadest spectrum of action, with therapeutic activity against dermatophytes, yeasts, and some nondermatophyte molds . Tissue pharmacokinetics accounts for significantly greater efficacy and much shorter treatment courses for fungal infections of the skin and nails . In general, oral itraconazole, fluconazole, and terbinafine are very well tolerated . The newer oral agents offer improved efficacy over griseofulvin and ketoconazole for onychomycosis and dry tinea pedis.

Chem Biol, 1997 Jun, 4(6), 461 - 71
Methionine aminopeptidase (type 2) is the common target for angiogenesis inhibitors AGM-1470 and ovalicin; Griffith EC et al.; BACKGROUND: Angiogenesis, the formation of new blood vessels, is essential for tumor growth . The inhibition of angiogenesis is therefore emerging as a promising therapy for cancer . Two natural products, fumagillin and ovalicin, were discovered to be potent inhibitors of angiogenesis due to their inhibition of endothelial cell proliferation . An analog of fumagillin, AGM-1470, is currently undergoing clinical trials for the treatment of a variety of cancers . The underlying molecular mechanism of the inhibition of angiogenesis by these natural drugs has remained unknown . RESULTS: Both AGM-1470 and ovalicin bind to a common bifunctional protein, identified by mass spectrometry as the type 2 methionine aminopeptidase (MetAP2) . This protein also acts as an inhibitor of eukaryotic initiation factor 2alpha (elF-2alpha) phosphorylation . Both drugs potently inhibit the methionine aminopeptidase activity of MetAP2 without affecting its ability to block elF-2alpha phosphorylation . There are two types of methionine aminopeptidase found in eukaryotes, but only the type 2 enzyme is inhibited by the drugs . A series of analogs of fumagillin and ovalicin were synthesized and their potency for inhibition of endothelial cell proliferation and inhibition of methionine aminopeptidase activity was determined . A significant correlation was found between the two activities . CONCLUSIONS: The protein MetAP2 is a common molecular target for both AGM-1470 and ovalicin . This finding suggests that MetAP2 may play a critical role in the proliferation of endothelial cells and may serve as a promising target for the development of new anti-angiogenic drugs.

Lipids, 1997 Jun, 32(6), 605 - 10
Sesamol as an inhibitor of growth and lipid metabolism in Mucor circinelloides via its action on malic enzyme; Wynn JP et al.; Sesamol, a nonoil component of sesame seed oil, inhibited growth, fatty acid synthesis, and desaturation by Mucor circinelloides in vivo . Although sesamol also inhibited the growth of other fungi and yeasts, its effect on the lipid metabolism of M . circinelloides was exceptional . An enzymological study demonstrated that sesamol affected lipid synthesis primarily by the inhibition of malic enzyme activity, thereby limiting the NADPH supply for fatty acid synthesis and desaturation . Sesamol itself had no inhibitory effect on malic enzyme activity in vitro . A metabolite of sesamol is therefore probably responsible for the in vivo effects of sesamol on lipid metabolism.

J Clin Microbiol, 1997 Jun, 35(6), 1353 - 60
Detection and identification of fungal pathogens in blood by using molecular probes; Einsele H et al.; A PCR assay was developed for the detection and identification of Candida and Aspergillus species . The design of the oligonucleotide primer pair as well as the species-specific probes used for species identification was derived from a comparison of the sequences of the 18S rRNA genes of various fungal pathogens . The primers targeted a consensus sequence for a variety of fungal pathogens . The assay was tested for sensitivity and specificity with 134 fungal and 85 nonfungal isolates . To assess clinical applicability, 601 blood samples from four defined groups were tested: group A (n = 35), controls; groups B to D (n = 86), patients with febrile neutropenia, without fungal colonization (group B; n = 29) and with fungal colonization (group C; n = 36); and patients with documented invasive fungal infection (IFI) (group D; n = 21) . The assay detected and, by species-specific hybridization, identified most of the clinically relevant Candida and Aspergillus species at 1 CFU/ml of blood . Amplification was 100% sensitive for all molds and yeasts tested, with Histoplasma capsulatum being the only non-Aspergillus species hybridizing with the Aspergillus spp . probe . None of 35 group A patients and only 3 of 65 group B and C patients were PCR positive . The sensitivity of the assay for specimens from patients with IFI (21 patients in group D) was 100% if two specimens were tested . For specificity, 3 of 189 specimens from patients at risk but with negative cultures were positive by the assay, for a specificity of 98% . PCR preceded radiological signs by a median of 4 days (range, 4 to 7 days) for 12 of 17 patients with hepatosplenic candidiasis or pulmonary aspergillosis . For the 10 patients with IFI responding to antifungal therapy, PCR assays became persistently negative after 14 days of treatment, in contrast to the case for 11 patients, who remained PCR positive while not responding to antifungal therapy . Thus, the described PCR assay allows for the highly sensitive and specific detection and identification of fungal pathogens in vitro and in vivo . Preliminary data from the screening of a selected group of patients revealed some value in the early diagnosis and monitoring of antifungal therapy.

Curr Opin Genet Dev, 1997 Apr, 7(2), 220 - 32
Life and death in the cytoplasm: messages from the 3' end; Wickens M et al.; The cytoplasmic life of an mRNA revolves around the regulation of its localization, translation and stability . Interactions between the two ends of the mRNA may integrate translation and mRNA turnover . Regulatory elements in the region between the termination codon and poly(A) tail - the 3' untranslated region - have been identified in a wide variety of systems, as have been some of the key players with which these elements interact.

Genes Dev, 1997 Mar 1, 11(5), 640 - 53
ALY, a context-dependent coactivator of LEF-1 and AML-1, is required for TCRalpha enhancer function; Bruhn L et al.; LEF-1 is a transcription factor that participates in the regulation of the T-cell receptor alpha (TCR alpha) enhancer by facilitating the assembly of multiple proteins into a higher order nucleoprotein complex . The function of LEF-1 is dependent, in part, on the HMG domain that induces a sharp bend in the DNA helix, and on an activation domain that stimulates transcription only in a specific context of other enhancer-binding proteins . With the aim of gaining insight into the function of context-dependent activation domains, we cloned ALY, a novel LEF-1-interacting protein . ALY is a ubiquitously expressed, nuclear protein that specifically associates with the activation domains of LEF-1 and AML-1 (CBF alpha2, PEBP2 alpha(B), which is another protein component of the TCR alpha enhancer complex . In addition, ALY can increase DNA binding by both LEF-1 and AML proteins . Overexpression of ALY stimulates the activity of the TCR alpha enhancer complex reconstituted in transfected nonlymphoid HeLa cells, whereas down-regulation of ALY by anti-sense oligonucleotides virtually eliminates TCR alpha enhancer activity in T cells . Similar to LEF-1, ALY can stimulate transcription in the context of the TCR alpha enhancer but apparently not when tethered to DNA through an heterologous DNA-binding domain . We propose that ALY mediates context-dependent transcriptional activation by facilitating the functional collaboration of multiple proteins in the TCR alpha enhancer complex.

Plant Physiol, 1997 Mar, 113(3), 943 - 9
Binding of the peroxisomal targeting sequence SKL is specified by a low-affinity site in castor bean glyoxysomal membranes . A domain next to the SKL binds to a high-affinity site; Wolins NE et al.; The carboxyl-terminal amino acid sequence serine-lysine-leucine (SKL) is the consensus peroxisomal targeting sequence 1 (PTS1) and is sufficient to direct a polypeptide to peroxisomes in vivo in plants, animals, and yeasts . However, there are also two sites on alkali-stripped glyoxysomal membranes from castor bean (Ricinus communis) endosperm that bind the peptide YHKHLKPLQSKL (SKLp), the sequence of the last 12 amino acids of acyl-coenzyme A oxidase (N.E . Wollins, R.P . Donaldson {1994} J Biol Chem 289: 1149-1153) . It was hypothesized that one of these sites interacts with information other than the PTS1 . To explore the sequence requirements for each SKLp binding site, we tested the peptides YHKHLKPQSKG and YHKHLKPLQS and found that they bound to the high-affinity site, but not to the low-affinity site . When the high-affinity site was blocked with YHKHLKPQSKG, SKLp bound to the low-affinity site with a dissociation constant (Kd) of 8.5 microM . In an attempt to disrupt high-affinity binding, two the upstream, positively charged residues were replaced with negatively charged residues to make the peptide YHKETEPLQSKL . YHKETEPLQSKL did not bind to either site on the glyoxysomal membranes . These results indicate that the PTS1 binds to the low-affinity site and that the adjacent, positively charged domain binds to the high-affinity site.

J Immunol, 1997 Feb 15, 158(4), 1779 - 86
Activation of human macrophage fungistatic activity against Histoplasma capsulatum upon adherence to type 1 collagen matrices; Newman SL et al.; Human monocyte/macrophages (Mphi) were adhered to extracellular matrix proteins, and the intracellular growth of Histoplasma capsulatum (Hc) yeasts were quantified and compared with their growth in Mphi adhered to plastic . Freshly isolated monocytes and cultured monocyte/derived Mphi adhered to type 1 collagen gels, but not to nongelled collagen-, fibronectin-, laminin-, or vitronectin-coated surfaces, demonstrated significant fungistatic activity against Hc yeasts . Activation of Mphi developed immediately upon adherence to the collagen matrices (1 h) and did not require additional time in culture . In addition, many of the yeasts were digested by 24 h postinfection . Mphi adhered to collagen maintained their fungistatic activity for up to 4 days, during which time monolayers cultured on plastic were destroyed . Culture of Mphi in the presence of IFN-gamma or TNF-alpha for 24 h before infection did not augment the fungistatic activity of collagen-adherent Mphi . Likewise, culture of monocytes on collagen gels with IL-3, granulocyte-Mphi CSF (GM-CSF) or Mphi CSF (M-CSF) for 7 days did not enhance Mphi fungistatic activity above that obtained by monocytes cultured on collagen alone . The mechanism(s) of Mphi-mediated fungistasis was not associated with production of toxic oxygen radicals, nitric oxide, or the restriction of intracellular iron . However, experiments with horseradish peroxidase-labeled gold colloids and immunoelectron microscopy demonstrated that phagolysosomal fusion, which is minimal in Hc-infected Mphi adhered to plastic, is enhanced significantly at both 1 h and 24 h postinfection in Mphi adhered to collagen matrices . These data suggest that in vivo, matrix-bound Mphi may express a previously unrecognized antifungal activity that proceeds in the absence of exogenous cytokines and is mediated, in part, by overcoming the capacity of Hc yeasts to inhibit Mphi phagolysosomal fusion.

Eur J Epidemiol, 1997 Feb, 13(2), 235 - 8
Transmission of nocardiosis and molecular typing of Nocardia species: a short review; Provost F et al.; Nocardia species are ubiquitous in the environment and may be found in the soil . They are generally responsible for sporadic pulmonary diseases acquired by inhalation of spores, with secondary localizations in the central nervous system and subcutaneous tissues . There is no absolute evidence for person to person transmission . Presumptive outbreaks of nocardiosis were observed in immunocompromised patients, more frequently in kidney transplant patients than in cardiac transplant patients . Nocardia spp., being present in dust particles, closure and disinfection of the transplantation unit with formaldehyde arrested the sequence of cases of nocardiosis . The original sources of the Nocardia sp . remain doubtful . Other possible sources of contamination are other patients, medical staff and the hospital environment . The first studies of Nocardia spp . typing were based on the detection of extracellular antigens, on the susceptibility of actinomycete strains to killer yeasts, and on the biochemical profiles with fluorogenic substrate . The use of molecular typing techniques have given very promising results . Analysis of plasmid profiles is an interesting way to compare the identity of isolates, although the reliability of this method depends of the presence of plasmids in the isolates . Other typing methods, including analysis of restriction length fragment polymorphism of total DNA, ribosomal DNA fingerprinting, require further investigations to evaluate their discriminating power or to be easily interpretable, whereas a random amplified polymorphic DNA (RAPD) assay was successful for epidemiological purposes . Progress in epidemiological analysis of cases of nocardiosis will be consistent when an improved diagnosis of this infection (molecular and serological diagnosis) will be available, when the genetic diversity of Nocardia spp . isolates will be better known, and when molecular typing, that hold promise in complementing investigations of outbreak of these infections, will be systematically performed when an abnormal increase of cases of nocardiosis in a population with risk factors is observed.

Plant Cell, 1997 Feb, 9(2), 223 - 35
Cell cycle phase specificity of putative cyclin-dependent kinase variants in synchronized alfalfa cells; Magyar Z et al.; The eukaryotic cell division cycle is coordinated by cyclin-dependent kinases (CDKs), represented by a single major serine/threonine kinase in yeasts (Cdc2/CDC28) and a family of kinases (CDK1 to CDK8) in human cells . Previously, two cdc2 homologs, cdc2MsA and cdc2MsB, have been identified in alfalfa (Medicago sativa) . By isolating cDNAs using a cdc2MsA probe, we demonstrate here that at least four additional cdc2 homologous genes are expressed in the tetraploid alfalfa . Proteins encoded by the new cdc2MsC to cdc2MsF cDNAs share the characteristic functional domains of CDKs with the conserved and plant-specific sequence elements . Transcripts from cdc2MsA, cdc2MsB, cdc2MsC, and cdc2MsE genes are synthesized throughout the cell cycle, whereas the amounts of cdc2MsD and cdc2MsF mRNAs peak during G2-to-M phases . The translation of Cdc2MsA/B, Cdc2MsD, and Cdc2MsF proteins follows the pattern of transcript accumulation . The multiplicity of kinase complexes with cell cycle phase-dependent activities was revealed by in vitro phosphorylation experiments . Proteins bound to p13suc1-Sepharose or immunoprecipitated with Cdc2MsA/B antibodies from cells at G1-to-S and G2-to-M phase boundaries showed elevated kinase activities . the Cdc2MsF antibodies separated a G2-to-M phase-related kinase complex . Detection of histone H1 phosphorylation activities in fractions immunoprecipitated with antimitotic cyclin (CyclinMs2) antibodies from G2-to-M phase cells indicates the complex formation between this cyclin and a kinase partner in alfalfa . The observed fluctuation of transcript levels, amounts, and activities of kinases in different cell cycle phases reflects a multilevel regulatory system during cell cycle progression in plants.

FEBS Lett, 1997 Jan 3, 400(2), 206 - 10
Identification of Asp804 and Asp808 as Na+ and K+ coordinating residues in alpha-subunit of renal Na,K-ATPase; Pedersen PA et al.; Mutations to Asp804 and Asp808 in the alpha-subunit almost abolish Na,K-ATPase activity, but high-affinity binding of {3H}ATP or {3H}ouabain at equilibrium and E1-E2 transitions are preserved . Titration of K+-ion displacement of {3H}ATP or {3H}ouabain shows that the mutations interfere with occlusion of K+ in the E2{2K} conformation . Reduced phosphorylation levels or affinities for Na+ in presence of oligomycin indicate that Asp804 and Asp808 also contribute to coordination of Na+ in the E1P{3Na} form . Demonstration of alternate interactions of Na+ or K+ with Asp804 and Asp808 support the notion of cation binding in a ping-pong sequence in catalytic models of Na,K-pumping.

Mycoses, 1997 Jan-Feb, 40(1-2), 3 - 24
Fungal infections of the cardiovascular system--a review; Kuttin ES; Various yeasts and filamentous fungal infections of human and animal heart and blood vessels, as well as the experimental investigations, are reviewed . Clinical, diagnostic, therapeutic and pathological aspects are discussed . The prevalence of mycotic infection of the heart, once considered to be an uncommon disease, has been reported frequently during recent years . The rate will certainly be higher if cases are more thoroughly investigated.

Biochem Cell Biol, 1997, 75(2), 95 - 102
Influence of the adenovirus 5 E1A oncogene on chromatin remodelling; Mymryk JS et al.; In the eukaryotic nucleus, compaction of DNA into chromatin can limit the access of trans-acting factors, providing an additional level of regulation to processes such as transcription, replication, and repair . Recent studies have suggested that the protein products of the adenovirus 5 E1A oncogene can influence SWI-SNF and histone acetylase activities, two cellular processes that facilitate transcription in the context of chromatin . This review focuses on the unexpected effects of E1A on cellular processes that remodel chromatin in relation to its transcriptional and transforming activities.

Adv Biochem Eng Biotechnol, 1997, 58, 185 - 230
Fluidized bed adsorption as a primary recovery step in protein purification; Thommes J; Fluidized bed adsorption has been introduced as an integrative technology combining clarification, concentration, and initial purification in a single step . In the paper presented here, the use of fluidized adsorbents in the primary recovery of proteins starting from unclarified broths is reviewed . First the principle of fluidizing adsorbent particles is discussed, subsequently possible experimental procedures for whole broth adsorption are demonstrated . The system parameters governing the performance of the sorption process in a fluidized bed are discussed in the second part of the paper and considerations on how operating parameters and process design influence the limiting steps are provided . Finally, examples for the successful operation of whole broth adsorption employing fluidized adsorbents are shown and conditions are defined under which this technology may be an alternative to traditional protein purification methods.

DNA Cell Biol, 1997 Jan, 16(1), 59 - 72
The transcriptional and translational control of diazepam binding inhibitor expression in rat male germ-line cells; Kolmer M et al.; The diazepam binding inhibitor {DBI, also known as acyl-CoA-binding protein, (ACBP), or endozepine} is a 10-kD protein that has been suggested to be involved in the regulation of several biological processes such as acyl-CoA metabolism, steroidogenesis, insulin secretion, and gamma-aminobutyric acid type A (GABA(A))/benzodiazepine receptor modulation . DBI has been cloned from vertebrates, insects, plants, and yeasts . In mammals, DBI is expressed in almost all the tissues studied . Nevertheless, DBI expression is restricted to specific cell types . Here we have studied DBI gene expression in the germ-line cells of rat testis . The DBI gene was intensively transcribed in postmeiotic round spermatids from stages VI to VIII of the seminiferous epithelial cycle . A prominent, spermatid-specific upstream transcription initiation site was identified in addition to the multiple common transcriptional initiation sites found in the somatic tissues . However, no DBI protein was detected in round spermatids, suggesting that the DBI transcripts were translationally arrested . The DBI protein was detected in the late spermatogenic stages starting from elongating spermatids from step 18 (stage VI) onward . The DBI protein was also detected in mature spermatozoa and in ejaculated human sperms . The majority of DBI was located at the middle piece of the spermatozoons tail enriched with mitochondria . On the basis of this observation and the well-established role of DBI in acyl-CoA metabolism, we propose that DBI expression in spermatozoa reflects the usage of fatty acids as a primary energy source by spermatozoa . The biological function of DBI in spermatozoa could thus be related to the motility function of sperm.

Bioessays, 1997 Jan, 19(1), 57 - 66
Peroxisome biogenesis; Waterham HR et al.; Peroxisomes are eukaryotic organelles that are the subcellular location of important metabolic reactions . In humans, defects in the organelle's function are often lethal . Yet, relative to other organelles, little is known about how cells maintain and propagate peroxisomes or how they direct specific sets of newly synthesized proteins to these organelles (peroxisome biogenesis/assembly) . In recent years, substantial progress has been made in elucidating aspects of peroxisome biogenesis and in identifying PEX genes whose products, peroxins, are essential for one or more of these processes . The most progress has been made in understanding the mechanism by which peroxisome matrix proteins are imported into the organelles . Signal sequences responsible for targeting proteins to the organelle have been defined . Potential signal receptor proteins, a receptor docking protein and other components of the import machinery have been identified, along with insights into how they operate . These studies indicate that multiple peroxisomal protein-import mechanisms exist and that these mechanisms are novel, not simply variations of those described for other organelles.

Leukemia, 1997 Jan, 11(1), 86 - 96
Elf-2, a rhombotin-2 binding ets transcription factor: discovery and potential role in T cell leukemia; Wilkinson DA et al.; Rhombotin-2 (RBTN-2) is a proto-oncogene only in the context of T lymphocytes . We postulated that the oncogenic effect of RBTN-2 in T cells is likely mediated by binding protein(s) with T cell-specific expression . By screening a T cell cDNA library, we identified a novel ets transcription factor that binds RBTN-2 . This protein was named elf-2 because its DNA-binding domain is virtually identical to that of ets family member elf-1 . Northern analyses showed similar levels of two elf-2 transcripts (3.5 kb and 3.8 kb) in all tissues except thymus . Thymocytes expressed four- to 10-fold greater amounts of the 3.5 kb transcript than other tissues . Sequence analyses of cDNA clones indicated that these transcripts encode proteins differing only at their amino termini, and likely represent alternatively spliced isoforms . These isoforms (elf-2a and elf-2b) contain identical RBTN-2 binding regions and DNA-binding domains . Elf-2b lacks a putative transactivation domain . The expression patterns suggest that RBTN-2 normally interacts equally with elf-2a and elf-2b . In contrast, when RBTN-2 is inappropriately expressed in T cells, RBTN-2 would interact predominantly with elf-2b; this interaction may lead to T cell proliferation.

Plant Cell, 1996 Dec, 8(12), 2223 - 34
PK12, a plant dual-specificity protein kinase of the LAMMER family, is regulated by the hormone ethylene; Sessa G et al.; The ethylene signal is transduced in plant cells via phosphorylation events . To identify protein kinases whose levels of expression are modulated by the plant hormone ethylene, we utilized a differential reverse transcriptase-polymerase chain reaction approach using mRNA extracted from ethylene-treated and untreated tobacco leaves . An ethylene-induced cDNA clone, PK12, encoding a protein kinase, was isolated . PK12 is a new member of the recently defined LAMMER family of protein kinases, which has been identified in mammals, flies, yeasts, and plants . The LAMMER kinases are related to the cell cycle-dependent CDC2-type kinases and are characterized by their similarity at kinase subdomain X . The recombinant PK12 protein autophosphorylates in vitro on serine, threonine, and tyrosine residues, thereby making it a member of the dual-specificity protein kinases . Immunoprecipitation of PK12 from plant extracts and kinase assay revealed that the apparent PK12 activity is rapidly and transiently increased when plants are treated with ethylene . By using in situ hybridization, we detected accumulation of the PK12 transcript in leaves after ethylene treatment and in the untreated flower abscission zone . The tissue in this zone is known to constitutively express ethylene-regulated genes.

FEMS Microbiol Lett, 1996 Dec 1, 145(2), 261 - 5
A method for the large scale isolation of high transformation efficiency fungal genomic DNA; Zhang D et al.; A procedure for isolation of genomic DNA from the zygomycete Cunninghamella elegans and other filamentous fungi and yeasts is reported . This procedure involves disruption of cells by grinding using dry ice, removal of polysaccharides using cetyltrimethylammonium bromide and by phenol extractions, and precipitation of DNA with isopropanol at room temperature . The isolation method produced large scale (approximate 1 mg DNA/5 g wet cells) and highly purified high molecular mass DNA . Sau3AI partially digested DNA showed high transformation efficiency (> 10(6)/100 ng DNA) when ligated to ZAP-express lambda vector.

Biol Signals, 1996 Nov-Dec, 5(6), 301 - 8
Protozoan cell cycle control; Wong JT; Many genes belonging to the cyclin-dependent kinase (cdk) family have been isolated from protozoans . While their role in cell cycle has yet to be proven unequivocally, at least one cdk can complement the cdc2ts/cdc28ts mutants in yeasts . Among the interesting questions relating to cdks in protozoa are: whether one cdk acts throughout the whole cell cycle and whether cyclin partnership is absolutely required . In protozoa, cell cycle control is closely associated with developmental control . Many life cycle differentiation phases can only occur during a specific window in the cell cycle . Because of different DNA biosynthetic pathways, some protozoa among the earliest eukaryotic lineages are unresponsive to common inhibitors of DNA synthesis like hydroxyurea . However, many protozoa do have different checkpoint controls in relation to their response to cell cycle inhibitors.

J Med Vet Mycol, 1996 Nov-Dec, 34(6), 371 - 8
Cell-mediated immunity in host resistance against infection caused by Penicillium marneffei; Kudeken N et al.; Penicillium marneffei is one of the most important opportunistic infectious pathogens in AIDS patients in Thailand and Southeast Asia . However, very little is known about the host defence mechanisms against P . marneffei infection . In the present study, we established the first experimental murine model of chronic pulmonary and disseminated infection using P . marneffei, and examined the immunological response to such infection in euthymic and athymic mice . In this model, micro-organisms inoculated intratracheally multiplied progressively in the lungs and disseminated to the liver and spleen . However, the number of organisms decreased gradually in these organs . In contrast, congenitally athymic mice developed severe pulmonary and disseminated systemic mycosis . Pulmonary penicilliosis marneffei was associated with a marked cellular inflammatory response as evident by histological abnormalities and increased intraparenchymal leucocyte count . To confirm the importance of cell-mediated immunity in host resistance to P . marneffei infection, we transferred nylon wool non-adherent spleen cells into the athymic mice . Such treatment significantly reduced the number of yeasts in the organs of athymic mice . Taken together, our results demonstrate that the cell-mediated immunity play a central role in a host defence mechanism against infection with P . marneffei, and suggest that our new model may be a useful approach for studying the pathogenesis of this fungal disease.

Cell, 1996 Nov 1, 87(3), 577 - 88
Crystal structure at 2.4 angstroms resolution of the complex of transducin betagamma and its regulator, phosducin; Gaudet R et al.; The crystal structure of transducin's betagamma subunits complexed with phosducin, which regulates Gtbetagamma activity, has been solved to 2.4 angstroms resolution . Phosducin has two domains that wrap around Gtbetagamma to form an extensive interface . The N-terminal domain binds loops on the "top" Gtbeta surface, overlapping the Gtalpha binding surface, explaining how phosducin blocks Gtbetagamma's interaction with Gtalpha . The C-terminal domain shows structural homology to thioredoxin and binds the outer strands of Gtbeta's seventh and first blades in a manner likely to disrupt Gtbetagamma's normal orientation relative to the membrane and receptor . Phosducin's Ser-73, which when phosphorylated inhibits phosducin's function, points away from Gtbetagamma, toward a large flexible loop . Thus phosphorylation is not likely to affect the interface directly, but rather indirectly through an induced conformational change.

J Biol Chem, 1996 Oct 25, 271(43), 26868 - 75
PIG-A and PIG-H, which participate in glycosylphosphatidylinositol anchor biosynthesis, form a protein complex in the endoplasmic reticulum; Watanabe R et al.; Many eukaryotic cell surface proteins are bound to the membrane via a glycosylphosphatidylinositol (GPI) anchor . Assembly of the GPI anchor precursor is a sequential addition of components to phosphatidylinositol (PI) in the endoplasmic reticulum (ER) . The first step is the transfer of N-acetylglucosamine (GlcNAc) to PI from UDP-GlcNAc to generate GlcNAc-PI . This simple step, however, is regulated by at least three genes because in both mammals and yeasts, there are three mutants of different complementation classes . To clarify this complexity, we analyzed the products of two cloned human genes, PIG-A and PIG-H . Here we demonstrate 1) that PIG-A is an ER transmembrane protein with a large cytoplasmic domain that has homology to a bacterial GlcNAc transferase and a small lumenal domain; 2) that PIG-H is a cytoplasmically oriented, ER-associated protein; and 3) that they form a protein complex . We also show that part of the small lumenal domain of PIG-A plays an essential functional role in targeting itself to the rough ER . Taken together with the cytoplasmic orientation of GlcNAc-PI, these results indicated that PIG-A and PIG-H are subunits of the GPI GlcNAc transferase that transfers GlcNAc to PI on the cytoplasmic side of the ER.

Biochim Biophys Acta, 1996 Oct 17, 1297(2), 235 - 43
Further studies on the bioaffinity chromatography of NAD(+)-dependent dehydrogenases using the locking-on effect; O'Carra P et al.; Previous studies have capitalized on ordered kinetic mechanisms in the design of biospecific affinity chromatographic methods for highly efficient purifications and mechanistic studies of enzymes . The most direct tactic has been the use of immobilised analogues of the following, usually enzyme-specific substrates, e.g., lactate/pyruvate in the case of lactate dehydrogenase for which NAD+ is the leading substrate . Such immobilised specific substrates are, however, often difficult or impossible to synthesise . The locking-on strategy reverses the tactic by using the more accessible immobilised leading substrate, immobilised NAD+, as adsorbent with soluble analogues of the enzyme-specific ligands (e.g., lactate in the case of lactate dehydrogenase) providing a substantial reinforcement of biospecific adsorption sufficient to effect adsorptive selection of an enzyme from a group of enzymes such as the NAD(+)-specific enzymes . The value of this approach is demonstrated using model studies with lactate dehydrogenase (LDH, EC 1.1.1.27), alcohol dehydrogenase (ADH, EC 1.1.1.1), glutamate dehydrogenase (GDH, EC 1.4.1.3) and malate dehydrogenase (MDH, EC 1.1.1.37) . Purification of bovine liver GDH in high yield from crude extracts is described using the tactic.

J Hosp Infect, 1996 Oct, 34(2), 123 - 9
Colonization and infection associated with Malassezia and Candida species in a neonatal unit; Shattuck KE et al.; The objectives of this study were to determine, in neonates of < 1250 g birthweight (N = 57), the initial time of skin colonization by Malassezia furfur, rate of colonization by Candida spp., and whether skin colonization by these yeasts was predictive of central line colonization or fungaemia . By age two weeks, 51% of neonates were culture-positive for M . furfur on umbilical or groin skin . During hospitalization, positive skin cultures for M . furfur or Candida spp . were obtained in 70% and 37% of neonates, respectively . Risk factors associated with positive skin cultures were mechanical ventilation and three or more episodes of suspected sepsis . Eight of the 52 infants with central venous catheters, had positive blood cultures withdrawn from the lines; five (62%) of these had positive skin surveillance cultures . Although positive skin cultures for M . furfur, Candida spp., or both were commonly observed in this population, they were not predictive of positive central line cultures or systemic illness.

J Biol Chem, 1996 Sep 6, 271(36), 22189 - 95
Regulation of the RNA polymerase I and III transcription systems in response to growth conditions; Clarke EM et al.; To better understand the mechanisms that regulate stable RNA synthesis, we have analyzed the RNA polymerase I and III transcriptional activities of extracts isolated from cells propagated under a variety of conditions . Under balanced growth conditions the levels of both RNA polymerase I- and III-specific transcription increased proportionally with growth rate . Upon nutritional starvation, RNA polymerase I transcription rapidly declined, followed by 5 S rDNA and eventually tDNA transcription . Transcriptional activities in extracts were restored when the nongrowing cultures were resuspended in fresh medium, although growth did not resume . The differential expression of 5 S rDNA and tDNA genes in extracts prepared from cells subjected to partial starvation was traced to a 5 S rDNA-specific inhibitor and not to a defect in any RNA polymerase III transcription factor . Characterization of this inhibitor indicated that it was not 5 S rRNA . It was sensitive to phenol extraction and resistant to RNase, and its target did not appear to be transcription factor IIIA . Not all treatments that slowed or stopped growth down-regulated the stable RNA transcription apparatus . Cells that have been subjected to either energy starvation or cycloheximide treatment still retain the ability to synthesize stable RNA in vitro, suggesting the presence of alternative regulatory mechanisms.

Biochim Biophys Acta, 1996 Sep 5, 1296(2), 145 - 51
Molecular characterisation of a thermoactive beta-1,3-glucanase from Oerskovia xanthineolytica; Parrado J et al.; Molecular characterisation of a lytic thermoactive beta-1,3-glucanase from Oerskovia xanthineolytica LL-G109 has been performed . A molecular mass of 27 195.6 +/- 1.3 Da and an isoelectric point of 4.85 were determined by electrospray mass spectrometry and from its titration curve, respectively . Its thermoactivity profile shows it to be a heat-stable enzyme with a temperature optimum of 65 degrees C . The secondary structure content of the protein was estimated by circular dichroism to be approx . 25% alpha-helix, 7% random coil, and 68% beta-sheet and beta-turn structure . Nuclear magnetic resonance spectra confirm the high content of beta-structure . Furthermore, the presence of a compact hydrophobic core is indicated by the presence of slowly exchanging amide hydrogens and the enzyme's relatively high resistance to proteolysis . The N-terminal sequences of the intact protein and of a tryptic peptide each exhibit significant similarity to family 16 of glycosyl hydrolases whose overall fold is known to contain almost exclusively beta-sheets and surface loops . Moreover, the sequenced tryptic peptide appears to encompass residues of the Oerskovia xanthineolytica glucanase active site, since it contains a portion of the family 16 active-site motif E-{L/I/V}-D-{L/I/V}-E.

Ann Pathol, 1996 Sep, 16(4), 266 - 70
{Role of guided fine needle punction in the diagnosis of deep-seated Aids-related infections . Report of a case of hepato-nodal histoplasmosis}; Longchampt E et al.; A case of disseminated histoplasmosis diagnosed by fine needle aspiration biopsy is reported . The patient suffering from acquired immune deficiency syndrome (AIDS) had enlarged liver, spleen and mesenteric lymph nodes . Cytological smears prepared from a CT scan guided fine needle aspiration biopsy of one of the lymph node and the liver, showed numerous free or intrahistiocytic yeasts consistent with Histoplasma capsulatum . Yeasts and protozoars morphologically close to Histo-plasma capsulatum are reviewed . The indications of fine needle aspiration biopsy for the diagnosis of infections in AIDS patients are emphasized . This method enables to send rapidly material for cultures and to start immediately an appropriate treatment.

J Enzyme Inhib, 1996 Aug, 11(1), 51 - 66
Piperastatin B: a new selective serine carboxypeptidase inhibitor from Streptomyces lavendofoliae MJ908-WF13; Murakami S et al.; Piperastatin B, a new inhibitor of serine carboxypeptidase was purified from a culture broth of Streptomyces lavendofoliae MJ908-WF13 as a minor component by monitoring its inhibitory activity against carboxypeptidase Y (CP-Y) . Its structure was determined to be N-formyl-Val-Thr-Leu-Val-Pip-Leu-Pip (pip: piperazic acid, hexahydropyridadine-3-carboxylic acid) . Piperastatin B is a highly specific competitive inhibitor of CP-Y (Ki = 55 nM) with little effect on related enzymes and resembles the major component, piperastatin A, in these respects.

Pediatr Allergy Immunol, 1996 Aug, 7(3), 151 - 5
Domestic fungal viable propagules and sensitization in children with IgE mediated allergic diseases; Dill I et al.; In order to study the possible relationship between domestic fungal exposure and sensitization to fungi, the homes of 20 children with allergic diseases (mainly bronchial asthma) were inspected for visible fungal growth and airborne viable fungal propagules were sampled . Inclusion criterion was sensitization to at least one of the tested fungi as shown by proof of specific IgE in serum . Twelve of 19 homes (63%) revealed a considerable amount of viable fungal propagules and/or visible fungal growth . Penicillium, Cladosporium, Aspergillus, Mycelia sterilia and yeasts were most prevalent . There was no clear-cut relationship to serologically proven sensitization.

Clin Infect Dis, 1996 Aug, 23(2), 305 - 13
Update on the management of onychomycosis: highlights of the Third Annual International Summit on Cutaneous Antifungal Therapy; Elewski BE et al.; Onychomycosis is an increasingly common fungal infection of the nail, which has traditionally been difficult to diagnose and treat and has physical and psychological consequences for the patient . Onychomycosis can be caused by dermatophytes, nondermatophytic filamentous fungi, and yeasts . The relative percentages of cases due to these etiologic agents vary with geographic location; however, in the United States, dermatophytes are the most common pathogens . Toenails are affected four times as often as fingernails . Microscopy and culture are the diagnostic "gold standards" for onychomycosis, although biopsy of the nail may be required to obtain a definitive diagnosis when conditions that mimic onychomycosis, such as psoriasis, are suspected . The treatment of onychomycosis includes a combination of topical therapy, surgical or chemical nail avulsion, and systemic therapy . The new generation of systemic agents (itraconazole, fluconazole, and terbinafine) is associated with a higher cure rate and shorter courses of treatment than are the older systemic antifungal drugs (i.e., griseofulvin and ketoconazole); these characteristics have sparked new interest in onychomycosis . Of these newer antifungals, itraconazole and terbinafine are the only agents currently approved by the U.S . Food and Drug Administration for the treatment of onychomycosis.

Genes Dev, 1996 Aug 1, 10(15), 1858 - 69
The EVES motif mediates both intermolecular and intramolecular regulation of c-Myb; Dash AB et al.; The c-Myb transcription factor is a proto-oncoprotein whose latent transforming activity can be unmasked by truncation of either terminus . Because both ends of Myb are involved in negative regulation, we tested whether they could associate in a two-hybrid assay and identified a carboxy-terminal motif that interacts with the amino-terminal DNA-binding domain . The EVES motif is highly conserved in vertebrate c-Myb proteins and contains a known site of phosphorylation previously implicated in the negative regulation of c-Myb . Interestingly, a related EVES motif is present in p100, a ubiquitously expressed transcriptional coactivator found in diverse species . We show that p100 interacts with and influences the activity of c-Myb, implicating it in the regulation of c-Myb, differentiation, and cell growth . Our results suggest that Myb is regulated by a novel mechanism in which intramolecular interactions and conformational changes control the intermolecular associations among Myb, p100, and the transcriptional apparatus.

Mol Cell Biol, 1996 Aug, 16(8), 4486 - 94
A consensus motif in the RFX DNA binding domain and binding domain mutants with altered specificity; Emery P et al.; The RFX DNA binding domain is a novel motif that has been conserved in a growing number of dimeric DNA-binding proteins, having diverse regulatory functions, in eukaryotic organisms ranging from yeasts to humans . To characterize this novel motif, we have performed a detailed dissection of the site-specific DNA binding activity of RFX1, a prototypical member of the RFX family . First, we have performed a site selection procedure to define the consensus binding site of RFX1 . Second, we have developed a new mutagenesis-selection procedure to derive a precise consensus motif, and to test the accuracy of a secondary structure prediction, for the RFX domain . Third, a modification of this procedure has allowed us to isolate altered-specificity RFX1 mutants . These results should facilitate the identification both of additional candidate genes controlled by RFX1 and of new members of the RFX family . Moreover, the altered-specificity RFX1 mutants represent valuable tools that will permit the function of RFX1 to be analyzed in vivo without interference from the ubiquitously expressed endogenous protein . Finally, the simplicity, efficiency, and versatility of the selection procedure we have developed make it of general value for the determination of consensus motifs, and for the isolation of mutants exhibiting altered functional properties, for large protein domains involved in protein-DNA as well as protein-protein interactions.

Chemotherapy, 1996 Jul-Aug, 42(4), 294 - 307
Antiproliferative effects of delta 24(25) sterol methyl transferase inhibitors on Trypanosoma (Schizotrypanum) cruzi: in vitro and in vivo studies; Urbina JA et al.; We have studied the antiproliferative effects of two sterol analogs previously reported as potent inhibitors of delta 24(25) sterol methyl transferase (E.C . 2.1.1.43) of yeasts and fungi on epimastigotes and amastigotes on Trypanosoma (Schizotrypanum) cruzi, the causative agents of Chagas disease, as well as its chemotherapeutic effects in a murine model of the disease . On the epimastigote form proliferating in liver infusion tryptose medium at 28 degrees C 22,26-azasterol (AZA), a cholestanol analog with a 6-membered aza ring as a side chain produced a dose-dependent reduction of the growth rate up to 3 microM, but at 10 microM complete growth arest and cell lysis took place after 120-144 h . For 24(R,S),25-epiminolanosterol (EIL), complete growth arrest and lysis took place with 6 microM . In both cases the antiproliferative effects were potentiated by the simultaneous incubation of the epimastigotes with inhibitors of sterol C-14 alpha-demethylase such as ketoconazole or SDZ 89,485, as indicated by concave isobolograms and fractional inhibitory concentrations ranging from 0.11 to 0.46 . Analysis of the sterol composition in control and treated cells by thin-layer and capillary gas-liquid chromatography coupled to mass spectrometry showed that growth inhibition correlated with the complete disappearance of the native endogenous sterols of the parasite (ergosterol and 24-ethyl analogs) and the accumulation of 24-desalkyl sterols . Against the clinically relevant amastigote form proliferating inside cultured Vero cells at 37 degrees C, AZA eradicated the parasite of 100 nM, while the corresponding concentration for EIL was 300 nM . Synergic effects of both inhibitors when combined with ketoconazole against this form of the parasite was demonstrated using a three-dimensional analytic method which allowed the identification of optimal drug concentrations . Finally, it was found that daily oral administration of AZA at 50 mg/kg/day for a total of 43 doses to mice infected with a lethal inoculum of T . cruzi allowed survival of all treated animals 25 days after infection, while all control (untreated) animals were dead at this point of time . Increased survival correlated with a 90% reduction in parasitemia in the treated animals . The antiparasitic effects of the azasterol were potentiated in combined treatments with ketoconazole . This is the first report of a successful application of a sterol methyl transferase inhibitor as a chemotherapeutic agent in a protozoal infection.

Oncogene, 1996 Jun 6, 12(11), 2291 - 300
A 15 amino acid stretch close to the Grb2-binding domain defines two differentially expressed hSos1 isoforms with markedly different Grb2 binding affinity and biological activity; Rojas JM et al.; We compared structure, expression and functional properties of two hSos1 cDNA isoforms (IsfI and Isf II) isolated, respectively, from human fetal brain and adult skeletal muscle libraries . IsfI and IsfII nucleotide sequences differ only by the presence in IsfII of an inframe 45 hp insertion located near the first proline-rich motif required for Grb2 binding . Some human tissues express only one isoform whereas others express different proportions of both in fetal and adult stages . In vitro binding assays and in vivo functional studies showed that MI exhibits significantly higher Grb2 binding affinity and biological activity than IsfI . These results suggest that functionally different hSos1 isoforms, with differential tissue expression and distribution, play important regulatory roles in the mechanisms controlling Ras activation in different tissues and/or developmental stages.

Mech Dev, 1996 Jun, 57(1), 3 - 20
Five years on the wings of fork head; Kaufmann E et al.; Since its discovery five years ago the conserved family of fork head/HNF-3-related transcription factors has gained increasing importance for the analysis of gene regulatory mechanisms during embryonic development and in differentiated cells . Different members of this family, which is defined by a conserved 110 amino acid residues encompassing DNA binding domain of winged helix structure, serve as regulatory keys in embryogenesis, in tumorigenesis or in the maintenance of differentiated cell states . The purpose of this review is to summarize the accumulating amount of data on structure, expression and function of fork head/HNF-3-related transcription factors.

Mol Endocrinol, 1996 Jun, 10(6), 694 - 704
Estrogen receptor affinity and location of consensus and imperfect estrogen response elements influence transcription activation of simplified promoters; Nardulli AM et al.; We have examined the ability of estrogen receptor (ER) to bind and bend DNA fragments containing the Xenopus laevis vitellogenin A2 estrogen response element (ERE), which contains a palindromic, consensus ERE sequence, the X . laevis vitellogenin B1 ERE2, which contains a 1-bp mismatch in the 5'-end of the half-palindrome, and the human pS2 ERE, which contains a 1-bp mismatch in the 3'-end of the half-palindrome . ER binding induced a 65 degrees bend in DNA fragments containing the consensus ERE, the vitellogenin B1 ERE2, or the pS2 ERE . However, ER affinity for the consensus ERE was 2-fold greater than for either the vitellogenin B1 ERE2 or the pS2 ERE . When Chinese hamster ovary (CHO) cells were transfected with reporter plasmids containing either the consensus ERE, the vitellogenin B1 ERE2, or the pS2 ERE separated from the TATA sequence by 26 helical turns, exposure to 10 nm 17 beta-estradiol increased transcription 12.7-, 2.4-, and 3.8-fold, respectively . Increasing the spacing between the ERE and TATA sequence to three helical turns decreased the ability of the consensus ERE to activate transcription by 55% and increased the ability of the pS2 ERE to activate transcription by 35% but had no significant effect on vitellogenin B1 ERE2 activity . Further increasing the distance between the ERE and TATA sequence to 3.6 helical turns restored the activity of promoters containing the consensus ERE and pS2 ERE but decreased the activity of the promoter containing the relatively weak vitellogenin B1 ERE2 . These data support the idea that 1) the affinity of ER for the ERE, 2) the location of an ERE within the promoter, and 3) the magnitude and orientation of DNA bends induced by binding of ER or other proteins are important in transcription activation of estrogen-responsive genes.

Rinsho Byori, 1996 Jun, 44(6), 512 - 7
{The antigen (CANDTEC antigen) detected by CAND TEC test for diagnosis of candidiasis}; Mitsuya M et al.; Most guinea pigs inoculated with 5.4 x 10(9) of C . albicans intraperitoneally, produce CANDTEC antigen (GPCANDTECAG) in sera . The antigen is heat-labile (at 56 degrees C for 30 min) as is that in humans . According to gel filtration, the molecule size of the antigen from guinea pigs was 4000KDa or more . ELISA revealed the antigen-positive gel fractions to contain a small amount of mannan from the yeasts and C3 . ELISA using rabbit anti-GPCANDTECAG serum indicated that the two CANDTEC antigens from guinea pigs and humans shared determinants . Gel filtration indicated that the CANDTEC antigen from patients was from 4000KDa to 3000KDa . In the antigen-positive gel fractions, IgM was detected by ELISA, but mannan and C3 were not detected . However, immunoblotting analysis on the antigen-positive fraction revealed a unique band of 200KDa, stained with concanavalin A-ALP . These findings indicate that CANDTEC antigens in guinea pigs and humans are immune complexes formed after infection of Candida, although the antigens have different components.

J Clin Microbiol, 1996 Jun, 34(6), 1583 - 5
Duration of incubation of fungal cultures; Morris AJ et al.; To determine the optimum duration of incubation for recovery of fungi, the results of 2,173 consecutive clinical cultures were reviewed . Overall, 94% of fungal isolates were detected by day 7 and 98% were detected by day 14 . Yeasts were usually (98%) detected within the first week of incubation . Recovery of molds required more time, but 81% were detected by day 7 and more than 96% were detected by day 14.

Cas Lek Cesk, 1996 May 29, 135(11), 335 - 9
{Oncogenes and the malignancy process}; Mares J et al.; An important group of genes for the development of neoplastic diseases are, in addition to tumour suppressor genes, protooncogenes . The latter are highly preserved genes present in a similar sequence in the cell genomes of different species (yeasts - man) . They encode components of biochemical signalling pathways by which external mitotic signals stimulate cell proliferation and products which inhibit cell differentiation . The result of activation of protooncogenes into oncogenes (mutations, chromosomal rearrangements, amplifications, viral insertions, insertion mutagenesis) is in particular hyperstimulation of cells resulting in uncontrolled proliferation . Mutations are of the dominant type, elimination of one allele leads to the transformation of a protooncogene into an oncogene . Oncogenes are classified with regard to the transmission level of the mitogenic signal on which they act . Originally they were detected in the genome of oncogenic viruses . However, they do not form their constant and specific constituent, the virus acts as a vector which transmits cellular protooncogenes (or oncogenes) during the reproductive cycle from one cell to another . The activity of various types of oncogenes is the necessary prerequisite for the genesis and development of various neoplastic diseases . Detection of oncogene alterations provides in some instances important diagnostic, prognostic and therapeutic findings.

Biochem Biophys Res Commun, 1996 May 24, 222(3), 802 - 8
The polyether bistratene A activates protein kinase C-delta and induces growth arrest in HL60 cells; Griffiths G et al.; Bistratene A (BisA) induced growth arrest in G2/M in HL60 cells . In addition, BisA-treated cells (50 nM for 48 h) became adherent and expressed the adhesion molecule CD11c, but did not express the monocyte enzyme alpha-napthyl acetate esterase or phagocytose complement coated yeasts . BisA activated protein kinase C (PKC)-delta and induced translocation of PKC-delta to the nucleus . This suggests that activation of PKC-delta can induce growth arrest and cell adhesion, but is insufficient to mediate full differentiation of HL60 cells . BisA has potential as a new probe for determining the function of PKC isoenzymes, specifically PKC-delta.

FEBS Lett, 1996 May 20, 386(2-3), 255 - 9
ADP delivery from adenylate kinase in the mitochondrial intermembrane space to oxidative phosphorylation increases in the presence of macromolecules; Laterveer FD et al.; Macromolecules were added to isolated rat liver mitochondria to mimic cytosolic macromolecules and tested for their effects on the ADP delivery from adenylate kinase in the intermembrane space to oxidative phosphorylation . In the presence of 10% (w/v) dextran M20 or bovine serum albumin, approximately 60% of the maximal ADP flux from adenylate kinase to oxidative phosphorylation was not accessible to an extramitochondrial ADP scavenger . In the absence of macromolecules this was 34% . ADP determinations from incubations with macromolecules demonstrated the existence of flux-dependent ADP concentration gradients across the outer membrane which can be as high as 12 microM.

Mutat Res, 1996 May 15, 363(1), 67 - 75
Multiple nuclear localization signals in XPG nuclease; Knauf JA et al.; We report here evidence for the mechanism of nuclear localization of XPG nuclease in human cells . Several candidate nuclear localization signal (NLS) peptides have been proposed for XPG protein . We have identified XPG peptides containing functional NLS and a potential nuclear retention signal (NRS) using in situ immunofluorescene localization of transiently expressed beta-galactosidase fusion proteins . Two XPG regions with putative NLS {amino acid (AA) coordinates: NLS-B (AA 1057-1074) and NLS-C (AA 1171-1185)} were each shown to independently localize the beta-gal extensively (> 80%) to the nucleus of HeLa cells . The C-terminus peptide containing NLS-C, an NLS conserved evolutionarily between yeasts and humans, also directed sub-localization of beta-galactosidase to intranuclear foci reminiscent of native XPG protein, as well as to peri-nucleolar regions . Peptides in the putative XPG 'NLS domain' (AA approximately 1051-1185) apparently function in concert for nuclear localization and also for retention of XPG in nuclear matrix-associated foci . Evidence presented elsewhere (Park et al., 1995) indicates that the peptide containing NLS-C (AA 1146-1185) also regulates the dynamic localization of XPG in the nucleus following UV-irradiation.

Mycoses, 1996 May-Jun, 39(5-6), 195 - 9
Disseminated infection with Trichosporon asahii; Itoh T et al.; Trichosporon fungaemia and disseminated, purpuric, papular skin lesions developed on the head, trunk and extremities of a 5-year-old female with acute lymphocytic leukaemia . Histopathologically, the skin lesions demonstrated dermal budding yeasts . She died despite treatment with antifungal drugs . The isolate from the blood was further identified morphologically and physiologically as Trichosporon asahii, based on the revision of the genus Trichosporon by Gueho et al . (1992) . According to the new revision, T . asahii is the only taxon regularly involved in systemic mycoses, so that most of the isolates previously reported as T . beigelii (formerly, T . cutaneum) in human deep mycoses are now thought to belong to T . asahii.

Hepatogastroenterology, 1996 May-Jun, 43(9), 771 - 5
Focal lesion of African histoplasmosis presenting as a malignant gastric ulcer; Sanguino JC et al.; We describe the case of a localized lesion of African Histoplasmosis, presenting as a gastric ulcer, with radiological and endoscopic features suggesting malignancy, that was submitted to surgery . Histologic examination of the resected specimen revealed typical yeasts of Histoplasma duboisii . There where no clinical, radiological or endoscopic signs of disseminated disease and conventional antifungal therapy was not prescribed . The patient has been followed for 11 years now, without evidence of relapse . There are few reports of gastrointestinal Histoplasmosis, and even fewer specifically caused by H . Duboissii Previous descriptions of gastric ulcer in immunocompetent hosts are related to disseminated forms of American Histoplasmosis . Although focal digestive lesions have been found in African patients, there is no available data on the incidence of gastric ulcer as a presenting sign of the disease.

Horm Metab Res, 1996 May, 28(5), 223 - 6
Influence of zinc and selenium deficiency on parameters relating to thyroid hormone metabolism; Kralik A et al.; 48 weaned male Sprague-Dawley rats with an initial average body weight of 41 g were divided into 4 groups of 12 animals (zinc-deficient; zinc-adequate, pair-fed with zinc-deficient group; selenium-deficient; selenium-adequate) for 40 days . All groups were fed a semisynthetic diet with casein being the source of protein . In the selenium-deficient diet, there was a selenium concentration of 0.038 mg/kg . The other diets were supplemented with Na-selenite in order to adjust the selenium concentration to 0.3 mg/kg . In the zinc-deficient diet, there was a zinc concentration of 4.1 mg/kg . The zinc concentrations in the other diets were adjusted to 45 mg/kg by the addition of zinc-sulfate heptahydrate . Zinc-deficient rats were characterized by a markedly reduced alkaline phosphatase activity in their serum, whilst selenium-deficient rats showed a markedly reduced glutathione peroxidase in serum proving their respective zinc-deficient and selenium-deficient states . Zinc deficiency decreased concentrations of triiodothyronine (T3) and free thyroxine (fT4) in serum by approximately 30% when compared with zinc-adequate controls . The concentration of thyroxine (T4) in serum was not affected by zinc deficiency . Selenium-deficient animals had lower concentrations of T3 and T4 than selenium-adequate animals . The concentration of fT4 in serum was not affected by selenium deficiency . The activity of hepatic type I 5'deiodinase was decreased by 67% by zinc deficiency and by 47% by selenium deficiency compared to adequate controls . The study data show that both zinc and selenium deficiency affect the metabolism of thyroid hormones.

Antonie Van Leeuwenhoek, 1996 Apr, 69(3), 217 - 22
Stationary phase development of Trimmatostroma abietis; Figueras MJ et al.; Processes of anamorph cell replication in Trimmatostroma abietis are described . Growth and conidiation are delimited on the basis of morphological, ultrastructural and ecological criteria . Cellular expansion shifts from bidirectional intercalary in exponential phase cells to isodiametric in late stationary phase cells, in the latter case with endogenous asexual reproduction . Ultrastructural similarities to dothideaceous black yeasts are discussed.

Drugs, 1996 Apr, 51(4), 585 - 620
Itraconazole . A reappraisal of its pharmacological properties and therapeutic use in the management of superficial fungal infections; Haria M et al.; Itraconazole is an orally administered triazole antifungal agent . Its spectrum of activity includes dermatophyte, dimorphic and dematiaceous fungi, yeasts, and some moulds . In clinical trials, mycological cure was attained in approximately 70 to 80, > or = 70 and > or = 80% of patients with, respectively, fingernail and toenail onychomycosis (200 mg/day for 3 months), dermatophytosis (100 mg/day for 2 to 4 weeks) and vaginal candidiasis (400 mg/day for 1 day or 200 mg/day for 3 days) . Approximately 20 to 30% of patients with onychomycosis may relapse after completion of therapy; relapse rate data are limited for the other indications . Recently developed intermittent regimens of itraconazole (400 mg/day for 1 week per month for 3 to 4 months) appear to have similar efficacy to standard regimens in the treatment of onychomycosis . Shorter, higher dosage itraconazole treatment regimens (200 or 400 mg/day for 1 week) are also beneficial in dermatomycoses . Discrepancies and limitations of study design hamper conclusions about efficacy relative to other antifungal drugs . Newer intermittent and short course higher dosage itraconazole regimens have also not been evaluated in comparative studies . Available studies show that the efficacy of itraconazole appears to be greater than that of griseofulvin, but similar to or lower than that of terbinafine in patients with dermatophyte onychomycosis or cutaneous fungal infections . Moreover, the efficacy of itraconazole may be similar to or lower than that of fluconazole in the treatment of cutaneous mycoses . Comparative data from patients with acute vaginal candidiasis suggest that itraconazole is at least as effective as intravaginal clotrimazole and oral fluconazole, and superior to intravaginal econazole . These results require confirmation . Prescription-event monitoring data indicate that itraconazole is generally well tolerated . Gastrointestinal disturbances, dizziness and headache occur most commonly; liver toxicity has been rarely described . Its usefulness in some clinical situations may be limited because of its ability to interact with various therapeutic agents . In conclusion, itraconazole along with other established agents should be considered a first-line treatment for patients with extensive or recalcitrant cutaneous fungal infections, mixed dermatophyte and Candida onychomycosis or vaginal candidiasis . It is currently considered a second-line drug for dermatophyte onychomycosis; the use of newer intermittent itraconazole treatment regimens may, however, extend its role in the management of this condition . Although itraconazole offers greater benefit than conventional therapies (griseofulvin and ketoconazole) in terms of efficacy and tolerability, wider clinical experience is required to determine its merits relative to the newer agents, terbinafine and fluconazole.

Am J Hum Genet, 1996 Apr, 58(4), 694 - 702
Thiopurine S-methyltransferase deficiency: two nucleotide transitions define the most prevalent mutant allele associated with loss of catalytic activity in Caucasians; Tai HL et al.; The autosomal recessive trait of thiopurine S-methytransferase (TPMT) deficiency is associated with severe hematopoietic toxicity when patients are treated with standard doses of mercaptopurine, azathioprine, or thioguanine . To define the molecular mechanism of this genetic polymorphism, we cloned and characterized the cDNA of a TPMT-deficient patient, which revealed a novel mutant allele (TPMT*3) containing two nucleotide transitions (G460-->A and A719-->G) producing amino acid changes at codons 154 (Ala-->Thr) and 240 (Tyr--> Cys), differing from the rare mutant TPMT allele we previously identified (i.e., TPMT*2 with only G238-->C) . Site-directed mutagenesis and heterologous expression established that either TPMT*3 mutation alone leads to a reduction in catalytic activity (G460-->A, ninefold reduction; A719-->G, 1.4-fold reduction), while the presence of both mutations leads to complete loss of activity . Using mutation specific PCR-RFLP analysis, the TPMT*3 allele was detected in genomic DNA from approximately 75 percent of unrelated white subjects with heterozygous phenotypes, indicating that TPMT*3 is the most prevalent mutant allele associated with TPMT-deficiency in Caucasians.

Cell, 1996 Mar 22, 84(6), 853 - 62
Site-specific phosphorylation of IkappaBalpha by a novel ubiquitination-dependent protein kinase activity; Chen ZJ et al.; Signal-induced activation of the transcription factor NF-kappaB requires specific phosphorylation of the inhibitor IkappaBalpha and its subsequent proteolytic degradation . Phosphorylation of serine residues 32 and 36 targets IkappaBalpha to the ubiquitin (Ub)-proteasome pathway . Here we report the identification of a large, multisubunit kinase (molecular mass approximately 700 kDa) that phosphorylates IkappaBalpha at S32 and S36 . Remarkably, the activity of this kinase requires the Ub-activating enzyme (E1), a specific Ub carrier protein (E2) of the Ubc4/Ubc5 family, and Ub . We also show that a ubiquitination event in the kinase complex is a prerequisite for specific phosphorylation of IkappaBalpha . Thus, ubiquitination serves a novel regulatory function that does not involve proteolysis.

Science, 1996 Mar 15, 271(5255), 1533 - 9
Phosphoinositides as regulators in membrane traffic; De Camilli P et al.; Phosphorylated products of phosphatidylinositol play critical roles in the regulation of membrane traffic, in addition to their classical roles as second messengers in signal transduction at the cell surface . Growing evidence suggests that phosphorylation-dephosphorylation of the polar heads of phosphoinositides (polyphosphorylated inositol lipids) in specific intracellular locations signals either the recruitment or the activation of proteins essential for vesicular transport . Cross talk between phosphatidylinositol metabolites and guanosine triphosphatases is an important feature of these regulatory mechanisms.

Science, 1996 Mar 15, 271(5255), 1526 - 33
Coat proteins and vesicle budding; Schekman R et al.; The trafficking of proteins within eukaryotic cells is achieved by the capture of cargo and targeting molecules into vesicles that bud from a donor membrane and deliver their contents to a receiving department . This process is bidirectional and may involve multiple organelles within a cell . Distinct coat proteins mediate each budding event, serving both to shape the transport vesicle and to select by direct or indirect interaction the desired set of cargo molecules . Secretion, which has been viewed as a default pathway, may require sorting and packaging signals on transported molecules to ensure their rapid delivery to the cell surface.

Biochemistry, 1996 Mar 12, 35(10), 3141 - 6
Secondary structure and topology of a mitochondrial presequence peptide associated with negatively charged micelles . A 2D H-NMR study; Chupin V et al.; In this study the secondary structure and topology of the peptide, corresponding to the presequence of cytochrome oxidase subunit IV (p25) in a negatively charged membrane-mimetic environment, were assessed by circular dichroism and two-dimensional nuclear magnetic resonance . The micelles used consisted of dodecylphosphoglycol (DPG), a mild anionic detergent with a headgroup resembling that of phosphatidylglycerol . The secondary structure was analyzed by interresidue nuclear Overhauser enhancement measurements and chemical shifts of backbone protons . The data revealed alpha-helix formation of the peptide upon interaction with the micelles, both in the N- and in the C-terminal halves, which are separated from each other by the proline residue at position 13 . The topology of the peptide was studied by determining the effect of spin-labeled 12-doxylstearate on the line widths of the peptide proton resonances . This method revealed the insertion of hydrophobic residues of both the N- and the C-terminal halves of p25 into the hydrophobic environment of the micelles, demonstrating the orientation of the amphiphilic helix.

FEBS Lett, 1996 Mar 11, 382(1-2), 18 - 20
Rabbit translation elongation factor 1 alpha stimulates the activity of homologous aminoacyl-tRNA synthetase; Negrutskii BS et al.; Functional and structural sequestration of aminoacyl-tRNA has been recently found in eukaryotic cells and the aminoacyl-tRNA channeling has been suggested {B.S . Negrutskii et al., Proc . Natl . Acad . Sci . 91 (1994) 964-968}, but molecular details and mechanism of the process remained unclear . In this paper we have verified a possible interaction between rabbit aminoacyl-tRNA synthetase and homologous translation elongation factor 1 alpha (EF-1 alpha), the proteins which may play a role of sequential components involved in the transfer of the aminoacyl-tRNA along the protein synthetic metabolic chain . The stimulation of the phenylalanyl-tRNA synthetase activity by EF-1 alpha is found . The effect is shown to be specific towards the origin of tRNA and elongation factor molecules . The data obtained favor the direct transfer mechanism of the aminoacyl-tRNA channeling process during eukaryotic protein synthesis.

Lipids, 1996 Mar, 31(3), 253 - 9
Study of the delta 12-desaturase system of Lipomyces starkeyi; Lomascolo A et al.; The specific activity of the microsomal delta 12-desaturase system, which transforms oleic acid into linoleic acid, was about 16 pmol/min/mg protein . However, most of the total activity was nonsedimentable even after a 200000 x g centrifugation for 100 min . The study of various physicochemical parameters showed that this enzymatic complex, functioning optimally between pH 7 and 8, had low thermal stability . Ca2+ which may cause an aggregation of the microsomes, and Hg2+ completely inhibited the activity, whereas Mg2+, Mn2+, and Zn2+ were activators . The delta 12-desaturase system was relatively specific toward oleic acid, though isomers of this fatty acid also had an action, either as substrates or as competitive inhibitors, on the activity of the system . The study of the effect of the exogenous oleoyl-CoA and elaidoyl-CoA on the specific activity of the delta 12-desaturase system showed a preference toward oleoyl-CoA.

Vet Pathol, 1996 Mar, 33(2), 238 - 41
Visceral mycosis in Chinook salmon (Oncorhynchus tschawytscha) due to Sporobolomyces salmonicolor; Muench TM et al.; One-month-old Chinook salmon fry from a cold-water hatchery were presented live for euthanasia and necropsy . Gross lesions were emaciation in 90% of the fry and ascites and increased cutaneous pigmentation in the remaining 10% . A cause for the emaciation was not determined . Histologically, the fry with ascites and increased pigmentation had visceral mycosis with aerocystitis, myositis, peritonitis, and dermatitis . Sporobolomyces salmonicolor, a rare human pathogen, was isolated and identified in tissue sections from affected fry.

J Med Vet Mycol, 1996 Mar-Apr, 34(2), 145 - 7
Isolation of Malassezia sympodialis from feline skin; Bond R et al.; Carriage of Malassezia yeasts was investigated in 17 cats in two colonies using a lipid-supplemented culture medium . Malassezia pachydermatis was isolated from one cat . Lipid-dependent Malassezia yeasts with electrophoretic karyotypes consistent with M . sympodialis were isolated from all six cats in one group and from one of 11 in the second group . To our knowledge, this is the first report of the isolation of lipid-dependent yeasts from cats.

J Mol Evol, 1996 Feb, 42(2), 321 - 2
CUG codons in Candida spp.; Jukes TH et al.; Codon CUG is used for serine instead of for leucine, its usual assignment, in several yeasts of the genus Candida . We propose a series of steps for the reassignment, including disappearance of leucine CUG and its anticodon CAG, formation of a new serine tRNA, with anticodon CAG, from a duplication of the gene for serine tRNA (IGA), and then production of CUG codons by mutation at sites that are mostly "nonessential."

J Antimicrob Chemother, 1996 Feb, 37(2), 275 - 83
Relative growth measurement of Candida species in a single concentration of fluconazole predicts the clinical response to fluconazole in HIV infected patients with oral candidosis; Cartledge JD et al.; The growth of 113 Candida spp . isolated prospectively from 104 HIV-positive patients treated for thrush was determined using a single concentration of 3 mg/L (10(-5) M) fluconazole relative to growth in drug free medium . Using a receiver operator characteristic curve, a relative growth in fluconazole of > or = 88% best discriminated between isolates from 56 patients who responded to treatment with fluconazole and 48 who failed to respond to 7 days, treatment with at least 100 mg/d drug with a sensitivity of 98% and a specificity of 96% . When the isolates from the eight patients with mixed infections due to both resistant and susceptible yeasts were excluded, the sensitivity and specificity both reached 100% . Relative growth of Candida spp . in a single concentration of fluconazole can therefore be used to accurately predict the clinical response to standard fluconazole treatment of thrush in patients infected with HIV.

Br J Oral Maxillofac Surg, 1996 Feb, 34(1), 23 - 5
A comparative study of the efficacy of fluconazole and amphotericin B in the treatment of oropharyngeal candidosis in patients undergoing radiotherapy for head and neck tumours; Finlay PM et al.; Radiotherapy given during treatment of oral and pharyngeal malignancy is frequently associated with colonization of the oral mucosa by Candida species . Treatment of these infections has included topical and systemic agents . In the present study 73 patients with oropharyngeal candidosis were treated with either amphotericin B (10 mg lozenges, four times daily for 14 days, 36 patients) or fluconazole (50 mg daily for 7 days, 37 patients) . The yeasts most frequently isolated were C albicans and C glabrata . Clinical signs and symptoms showed improvement at end of treatment in 72% of patients who received amphotericin B compared with 92% of patients who received fluconazole . Mycological cure at end of treatment was achieved in 31% of the amphotericin B group and 46% of patients who received fluconazole . For both treatments the cure rate was less in denture wearers than in non denture wearers.

J Am Acad Dermatol, 1996 Feb, 34(2 Pt 1), 273 - 7
Present and potential diagnostic techniques in onychomycosis; Pierard GE et al.; The problem of onychomycosis has been frequently addressed during recent years . To make the diagnosis of onychomycosis dermatologists have relied on clinical presentation, culture, and microscopy . These approaches are hampered by false-negative and false-positive results that have confused treatment outcomes . Two new diagnostic techniques, immunohistochemistry and flow cytometry, provide an effective means of identifying different dermatophytes, yeasts, and nondermatophytic molds . Immunohistochemistry employs antibodies to certain fungi to enable positive identification in situ, whereas flow cytometry differentiates fungi on the basis of molecular differences . These techniques provide new evidence that nondermatophytic molds and yeasts can actively invade nail tissue and that mixed infections occur . These findings could have important implications for the treatment of onychomycosis.

Arch Dermatol, 1996 Feb, 132(2), 190 - 3
Neonatal Malassezia furfur pustulosis; Rapelanoro R et al.; BACKGROUND: Papulopustular eruptions of the face in neonates are frequently referred to as neonatal acne or sebaceous miliaria . Our findings suggest that there is an association between this type of eruption and Malassezia furfur infection . OBSERVATIONS: Direct examination of pustule smears showed M furfur yeasts in eight of 13 cases involving neonates with erythema and papulopustules of the face, neck, and scalp (mean age at onset, 22 days {range, 7 to 30 days}) . The pustules were predominantly neutrophilic . Treatment with 2% ketoconazole cream applied topically twice daily was effective in 1 week . CONCLUSION: Malassezia furfur is frequently associated with a common nonfollicular pustulosis of the newborn, probably improperly termed neonatal acne.

J Inorg Biochem, 1996 Feb, 61(2), 125 - 42
Ag(I)-binding to phytochelatins; Mehra RK et al.; Phytochelatins (PCs) are glutathione-derived peptides with the general structure (gamma-Glu-Cys)nGly, where n varies from 2 to 11 . A variety of metal ions such as Cu(II), Cd(II), Pb(II), Zn(II), and Ag(I) induce PC synthesis in plants and some yeasts . It has generally been assumed that the inducer metals also bind PCs . However, very little information is available on the binding of metals other than Cu(I) and Cd(II) to PCs . In this paper, we describe the Ag(I)-binding characteristics of PCs with the structure (gamma-Glu-Cys)2Gly, (gamma-Glu-Cys)3Gly, and (gamma-Glu-Cys)4Gly . The Ag(I)-binding stoichiometries of these three peptides were determined by (i) UV/VIS spectrophotometry, (ii) luminescence spectroscopy at 77 K, and (iii) reverse-phase HPLC . The three techniques yielded similar results . ApoPCs exhibit featureless absorption in the 220-340 nm range . The binding of Ag(I) to PCs induced the appearance of specific absorption shoulders . The titration end point was indicated by the flattening of the characteristic absorption shoulders . Similarly, luminescence at 77 K due to Ag(I)-thiolate clusters increased with the addition of graded Ag(I) equivalents . The luminescence declined when Ag(I) equivalents in excess of the saturating amounts were added to the peptides . At neutral pH, (gamma-Glu-Cys)2Gly, (gamma-Glu-Cys)3Gly, and (gamma-Glu-Cys)4Gly bind 1.0, 1.5, and 4.0 equivalents of Ag(I), respectively . The Ag(I)-binding capacity of (gamma-Glu-Cys)2Gly and (gamma-Glu-Cys)3Gly was increased at pH 5.0 and below so that Ag(I)/-SH ratio approached 1.0 . A similar pH-dependent binding of Ag(I) to glutathione was also observed . The increased Ag(I)-binding to PCs at lower pH is of physiological significance as these peptides accumulate in acidic vacuoles . We also report lifetime data on Ag(I)-PCs . The relatively long decay-times (approximately 0.1-0.3 msec) accompanied with a large Stokes shift in the emission band are indicative of spin-forbidden phosphorescence.

Tsitologiia, 1996, 38(9), 889 - 913
{The structural and functional characteristics of the dolichol cycle enzymes}; Shpakov AO et al.; The literature and the authors' our data on the biosynthesis of a dolichol derivative Dol-PP-GlcNAc2Man9Glc3, involved in protein N-glycosylation in the endoplasmic reticulum (ER) of the eukaryotic cells, have been summarized and analysed . The structural and functional characteristics of dolichol-coupled enzymes, catalyzing biosynthesis of Dol-PP-GlcNAc2Man9Glc3, are considered . It is shown that the dolichol cycle enzymes, in conformity with their structural peculiarities and ER membrane topology, can be divided into three groups having the common evolutionary origin . Possible mechanisms of the dolichol derivative translocation through membrane is discussed . A conclusion is made about formation of multicomponent complexes by dolichol-coupled enzymes . These complexes secure functional co-ordination of the enzymes.

Environ Mol Mutagen, 1996, 28(3), 176 - 81
Aneuploidy in germ cells: disruption of chromosome mover components; Preston RJ; The task of the Workgroup on "Disruption of Chromosome Mover Components" was to establish what cellular structures are involved in chromosome segregation and how disruption of these could occur . Recent research on the mechanism of action of the cellular components that segregate chromosomes accurately during mitosis or meiosis has served to highlight the number of potential targets for disruption . The process of chromosome segregation represents an orchestrated chain of events centered on the activities of cellular motors, kinesins and dyneins . These motors are involved in arranging chromosomes at the metaphase plate, providing the spindle tension necessary for progression, and the actual segregation of the chromosomes to the poles . The Workgroup determined that there is a lack of information on the effects of chemical exposure to cell motors and other chromosome mover components, and that there is a clear need for further research . This article describes the discussions of the Workgroup and highlights areas of future research into chromosome movement, particularly in human meiotic and mitotic cells . The Workgroup emphasized that obtaining mechanistic data on the induction of aneuploidy will allow for extrapolation of the dose response curves for chemical exposures below the level of observation and for using aneuploidy data for quantitative risk assessment for adverse health effects.

Adv Enzyme Regul, 1996, 36, 3 - 15
Differences in the properties of mammalian ribonucleotide reductase toward its substrates; Cory JG et al.; These studies, using three different reagents, show that the substrate properties of ribonucleotide reductase are specific but can be variable depending upon the nature of the interaction of the reagent with the holoenzyme or the individual subunit . Etheno-CDP, which acts as a competitive inhibitor with respect to CDP, interacts with the active site of the holoenzyme . This interaction was the result of rather tight structural requirements as epsilon-ADP did not result in a similar level of inhibition of either CDP or ADP reductase activities . The YL 1/2 antibody which binds very tightly to the NHI subunit has a much greater effect on CDP reductase activity than ADP reductase activity . The nonapeptide that corresponds to the C-terminus amino acid sequence of the NHI subunit and which binds to the EB subunit and aborts the formation of the NHI-EB active complex has a greater effect on ADP reductase activity than on CDP reductase activity . The use of reagents such as these can be helpful in dissecting the subtle but important differences in the substrate properties of mammalian ribonucleotide reductase.

EXS, 1996, 77, 307 - 20
Toxic metal-responsive gene transcription; Zhu Z et al.; Metals play a dual role in biological systems, serving as essential co-factors for a wide range of biochemical reactions yet these same metals may be extremely toxic to cells . To cope with the stress of increases in environmental metal concentrations, eukaryotic cells have developed sophisticated toxic metal sensing proteins which respond to elevations in metal concentrations . This signal is transmitted to stimulate the cellular transcriptional machinery to activate expression of metal detoxification and homeostasis genes . This review summarizes our current understanding of the biochemical and genetic mechanisms which underlie cellular responses to toxic metals via metalloregulatory transcription factors.

Rapid Commun Mass Spectrom, 1996, 10(11), 1371 - 8
Delayed extraction improves specificity in database searches by matrix-assisted laser desorption/ionization peptide maps; Jensen ON et al.; Peptide mass maps obtained by matrix-assisted laser desorption ionization (MALDI) are an attractive means to identify proteins by searches in sequence databases . Here we demonstrate that the recently introduced delayed ion-extraction technique, when coupled to reflectron MALDI time-of-flight mass spectrometry, leads to dramatically improved search specificity . Routine resolution in the range of 6,000 to 12,000 allows assignment of monoisotopic masses throughout the peptide mass range . Database searches can be performed with high precision by use of a mass accuracy which is currently better than 30 ppm over a wide mass range and better than 5 ppm for a narrow mass range . This high performance makes it possible to identify proteins with fewer peptide masses than before . Additional low intensity peaks can be assigned after a search because of the improved signal-to-noise ratio of delayed-extraction peptide mass spectra, increasing sequence coverage of matched proteins . The improvements in database search specificity can be used to identify the components of simple protein mixtures . In combination with advanced sample preparation and automation techniques, delayed-extraction MALDI time-of-flight mass spectrometry is now an extremely powerful tool for the database identification of proteins.

Ann Dermatol Venereol, 1996, 123(3), 157 - 64
{Allergologic survey in 251 patients with moderate or severe dermatitis . Incidence and value of the detection of contact eczema, food allergy or sensitization to air-borne allergens}; Guillet MH et al.; INTRODUCTION: Because of the increased recruitment of uncontrolled atopic dermatitis (AD) necessitating chronic use of dermocorticosteroids, we developed a prospective allergologic survey in a serie of 251 patients presenting with moderate or severe AD . PATIENTS AND METHOD: 251 patients were refered for allergologic assessment and followup . The clinical severity was assessed by use of standardized scores . Patients were grouped by age: group 1 (70 children younger than 2 years), group 2 (93 children between 2 and 7 years), group 3 (23 children between 7 and 15 years), group 4 (65 children over 15 years and adults) . All the patients were systematically screened for contact dermatitis and IgE mediated sensitization (inhallant and food allergens) with blood tests for IgE, prior to evaluation of clinical relevance . RESULTS: Aero-allergen sensitization was demonstrated in 51 p . 100 of children and 89 p . 100 of adults . It was present earlier in severe AD with main clinical involvement for nose and throat and respiratory symptoms . Clinical responsibility for dermatitis was documented in only 6 p . 100 of AD . Food allergy was early incriminated as flare factors in most of severe AD (96 p . 100 of children and 81 p . 100 of adults) with major and persistant improvement under eviction diet . Main allergens were eggs (46 p . 100), pea-nuts (29 p . 100), shellfish (24 p . 100), milk (20 p . 100), flour (14 p . 100), fish (14 p . 100), soybeans (8.9 p . 100) . Food allergy to yeasts (7.2 p . 100) was important in adults . Food allergy is the earliest allergy in the course of severe AD and the number of involved trophallergens increases in older patients . Patch tests were positive in 40 p . 100 of patients (i . e . 31 p . 100 of children and 66 p . 100 of adults) with a greater incidence in moderate AD . Main allergens were metals (54 p . 100), fragrances (19 p . 100), balsam of Peru (10 p . 100), parabens (8 p . 100) and lanoline (4 p . 100) . CONCLUSION: When AD is not efficiently controlled by dermocorticosteroids, allergologic screening and treatment of children and adults proves to be very interesting . Specific measures regarding food allergy and contact dermatitis reduce or vanish cutaneous flares . As for inhallant sensitizations, Dermatologists should be awared that they may play a role regarding assessment of sensitization and prevention of respiratory symptoms in moderate and severe AD since the risk of complications is important in both groups.

J Ethnopharmacol, 1995 Dec 15, 49(3), 163 - 9
Antifungal activity of turmeric oil extracted from Curcuma longa (Zingiberaceae); Apisariyakul A et al.; Turmeric oil and curcumin, isolated from Curcuma longa L., were studied against fifteen isolates of dermatophytes, four isolates of pathogenic molds and six isolates of yeasts . The inhibitory activity of turmeric oil was tested in Trichophyton-induced dermatophytosis in guinea pigs . The results showed that all 15 isolates of dermatophytes could be inhibited by turmeric oil at dilutions of 1:40-1:320 . None of the isolates of dermatophytes were inhibited by curcumin . The other four isolates of pathogenic fungi were inhibited by turmeric oil at dilutions of 1:40-1:80 but none were inhibited by curcumin . All six isolates of yeasts tested proved to be insensitive to both turmeric oil and curcumin . In the experimental animals, turmeric oil (dilution 1:80) was applied by dermal application on the 7th day following dermatophytosis induction with Trichophyton rubrum . An improvement in lesions was observed in 2-5 days and the lesions disappeared 6-7 days after the application of turmeric oil.

Curr Opin Cell Biol, 1995 Dec, 7(6), 781 - 9
The self-destructive personality of a cell cycle in transition; Deshaies RJ; The transition from G1 to S phase, sister chromatid separation in anaphase, and the exit from mitosis are driven by the destruction of cell cycle regulatory proteins by distinct ubiquitin-dependent proteolytic pathways . The components and targets of these key degradation pathways are now becoming clear . Genetic and biochemical dissections of these extremely specific and well regulated destruction pathways are providing fundamental insights into the mechanisms of control of the cell division cycle.

Cell, 1995 Dec 1, 83(5), 683 - 92
Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins; Rexach M et al.; The molecular dynamics of nuclear protein import were examined in a solution binding assay by testing for interactions between a protein containing a nuclear localization signal (NLS), the transport factors karyopherin alpha, karyopherin beta, and Ran, and FXFG or GLFG repeat regions of nucleoporins . We found that karyopherins alpha and beta cooperate to bind FXFG but not GLFG repeat regions . Binding of the NLS protein to karyopherin alpha was enhanced by karyopherin beta . Two novel reactions were discovered . First, incubation of a karyopherin heterodimer-NLS protein complex with an FXFG repeat region stimulated the dissociation of the NLS protein from the karyopherin heterodimer . Second, incubation of the karyopherin heterodimer with RanGTP (or with a Ran mutant that cannot hydrolyze GTP) led to the dissociation of karyopherin alpha from beta and to an association of Ran with karyopherin beta; RanGDP had no effect . We propose that movement of NLS proteins across the nuclear pore complex is a stochastic process that operates via repeated association-dissociation reactions.

FEBS Lett, 1995 Nov 27, 376(1-2), 91 - 4
Automation of micro-preparation and enzymatic cleavage of gel electrophoretically separated proteins; Houthaeve T et al.; To achieve high throughput, protein microcharacterization sample preparation must be automated . We describe a cartesian robot capable of processing 32 protein samples in parallel . The system is based on specially designed flow-through reactors for contamination-free reagent delivery and removal . Washing of excised gel pieces, reduction and alkylation, proteolytic cleavage and peptide extraction are performed in these reactors . Compatibility of the system with HPLC peptide separation and Edman degradation as well as with laser desorption mass spectrometry of the unseparated mixture is demonstrated . This is the first report describing automated preparation and processing of multiple protein samples.

Mol Gen Genet, 1995 Nov 15, 249(2), 185 - 90
Genetic mapping of mitochondrial markers by recombinational analysis in Chlamydomonas reinhardtii; Remacle C et al.; The mitochondrial genome of Chlamydomonas reinhardtii is a 15.8 kb linear DNA molecule present in multiple copies . In crosses, the meiotic products only inherit the mitochondrial genome of the mating type minus (paternal) parent . In contrast mitotic zygotes transmit maternal and paternal mitochondrial DNA copies to their diploid progeny and recombinational events between molecules of both origins frequently occur . Six mitochondrial mutants unable to grow in the dark (dk- mutants) were crossed in various combinations and the percentages of wild-type dk+ recombinants were determined in mitotic zygotes when all progeny cells had become homoplasmic for the mitochondrial genome . In crosses between strains mutated in the COB (apocytochrome b) gene and strains mutated in the COX1 (subunit 1 of cytochrome oxidase) gene, the frequency of recombination was 13.7% (+/- 3.2%) . The corresponding physical distance between the mutation sites was 4.3 kb . In crosses between strains carrying mutations separated by about 20 bp, a recombinational frequency of 0.04% (+/- 0.02%) was found . Two other mutants not yet characterized at the molecular level were also used for recombinational studies . From these data, a linear genetic map of the mitochondrial genome could be drawn . This map is consistent with the positions of the mutation sites on the mitochondrial DNA molecule and thereby validates the method used to generate the map . The frequency of recombination per physical distance unit (3.2% +/- 0.7% per kilobase) is compared with those obtained for other organellar genomes in yeasts and Chlamydomonas.

Cell, 1995 Nov 3, 83(3), 423 - 32
Rabaptin-5 is a direct effector of the small GTPase Rab5 in endocytic membrane fusion; Stenmark H et al.; We have identified a novel 100 kDa coiled-coil protein, rabaptin-5, that specifically interacts with the GTP form of the small GTPase Rab5, a potent regulator of endocytic transport . It is mainly cytosolic, but a fraction colocalizes with Rab5 to early endosomes . Expression of a GTPase-deficient Rab5 mutant enhances the binding of rabaptin-5 to enlarged endosomes . Overexpression of rabaptin-5 alone is sufficient to promote expansion of early endosomes . Rab5 recruits rabaptin-5 to purified early endosomes in a GTP-dependent manner, demonstrating functional similarities with other members of the Ras superfamily . Immunodepletion of rabaptin-5 from cytosol strongly inhibits Rab5-dependent early endosome fusion . Rabaptin-5 is thus a Rab effector required for membrane docking and fusion.

Pediatr Allergy Immunol, 1995 Nov, 6(4), 181 - 6
Viable fungi in indoor air in homes and schools in the Sør-Varanger community during winter; Dotterud LK et al.; The present study investigated the content of fungal aerospores in homes and schools of house-dust-mite (HDM)-sensitized and control children in a subarctic area . During winter, airborne microfungi were collected from the homes and schools of 19 HDM-sensitized children and 19 nonatopic controls, all living in the community of Sor-Varanger, northern Norway . The samples were cultivated and microfungal growth was identified microscopically . Indoor humidity, temperature, and carbon dioxide (CO2) concentrations were measured . Housing conditions and sociodemographic and symptom data were obtained by a questionnaire . Penicillium was the most common microfungus in both homes and schools, followed by various yeasts, Aspergillus, Cladosporium, and Mucor . The number of infected homes was equal in the HDM-sensitized group and the control group, but aerospore counts were higher in the HDM-sensitized group than in the control group . The lowest aerospore counts were found in the schools . High aerospore counts also appeared to be related to high indoor humidity . The keeping of pets and damp indoor conditions were more frequent in homes of HDM-sensitized children than in the control group, whereas parental smoking and carpeting occurred with equal frequency in both groups . This indicates that no allergy sanitation measures had been undertaken, especially in the homes of the HDM-sensitized children.

J Antibiot (Tokyo), 1995 Nov, 48(11), 1262 - 6
Chondramides A approximately D, new antifungal and cytostatic depsipeptides from Chondromyces crocatus (myxobacteria) . Production, physico-chemical and biological properties; Kunze B et al.; Novel depsipeptides, named chondramides were produced at levels up to 4.3 mg/liter by several myxobacteria of the genus Chondromyces . The compounds are structurally closely related to jaspamide/jasplakinolide from marine sponges of the genus Jaspis . Initially the chondramides were detected in acetone extracts of the biomass of Chondromyces crocatus, strain Cm c2 . So far, four structural variants could be characterized, the chondramides A approximately D . They inhibited the growth of a few yeasts and showed high cytostatic activity against cultivated human and animal cells.

J Med Microbiol, 1995 Nov, 43(5), 360 - 7
An alpha 5 beta 1-like integrin receptor mediates the binding of less pathogenic Candida species to fibronectin; Santoni G et al.; The present study was undertaken to investigate whether less pathogenic Candida species (C . tropicalis, C . stellatoidea, C . krusei and C . glabrata) express a fibronectin receptor (FNr) antigenically related to alpha 5 beta 1 integrin, which mediates their binding to fibronectin (FN) . By flow cytometric analysis, a monoclonal antibody (MAb) directed against human alpha 5 integrin subunit (clone SAM-1) and two different antisera to FNr positively stained C . tropicalis, C . stellatoidea and C . glabrata, with the greatest expression observed for C . tropicalis . No or only marginal immunoreactivity was found on C . krusei . C . tropicalis, C . stellatoidea, C . glabrata, but not C . krusei yeasts specifically adhered to FN; higher levels of adhesion were found for C . tropicalis and C . stellatoidea with respect to C . glabrata . Less pathogenic Candida spp . bound to the Arg-Gly-Asp (RGD) containing 120-kDa fragment of FN and adhesion to intact FN was markedly inhibited by Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), but not by Gly-Arg-Gly-Glu-Ser-Pro (GRGESP) peptides . In addition, anti-alpha 5 SAM-1 MAb and both anti-FNr antisera strongly blocked binding of less pathogenic Candida spp . to FN . Overall, these results indicate that less pathogenic Candida spp., including C . tropicalis, C . stellatoidea and C . glabrata, express a receptor antigenically related to alpha 5 beta 1 integrin which mediates their adhesion to FN.

Biochemistry, 1995 Oct 24, 34(42), 13934 - 42
Probing intradomain and interdomain conformational changes during equilibrium unfolding of phosphoglycerate kinase: fluorescence and circular dichroism study of tryptophan mutants; Sherman MA et al.; Phosphoglycerate kinase is a monomeric protein composed of two globular domains of the alpha/beta type . Extensive domain-domain interactions involve three segments of the polypeptide chain that are distant from one another in the primary sequence: the N-terminus, the C-terminus, and a centrally located alpha-helix . In order to monitor spectroscopically the conformational changes that occur in the individual domains and at the interdomain interface during the unfolding process, we have constructed a series of single-tryptophan mutants . In addition to two previously described mutants, each with single tryptophans in the C-terminal domain (W308 and W333) {Szpikowska, B . K., Beechem, J . M., Sherman, M . A., & Mas, M . T . (1994) Biochemistry 33, 2217-2225}, four new single-tryptophan mutants have been constructed: two with tryptophans located in the interdomain region (W194 and W399) and two with tryptophans in the N-terminal domain (W48 and W122) . The equilibrium unfolding transitions induced by guanidine hydrochloride were monitored using far-UV CD, near-UV CD, steady-state, and time-resolved fluorescence . These studies reveal two unfolding transitions and suggest a sequential unfolding process for the mutants described in this paper . During the first transition (Cm approximately 0.5 M) the interdomain region and C-terminal domain unfold; the N-terminal domain remains relatively compact but lacks much of the tertiary structure that characterizes the native state . A hyperfluorescent intermediate is detected during this transition by tryptophan probes placed within the N-terminal domain . Complete unfolding of the N-terminal domain occurs during the second transition (Cm approximately 0.9 M).

Trends Biochem Sci, 1995 Oct, 20(10), 405 - 11
Post-translational modification of poly(ADP-ribose) polymerase induced by DNA strand breaks; Lindahl T et al.; There are one million molecules of poly(ADP-ribose) polymerase (PARP) in mammalian cell nuclei and the enzyme is found in most eukaryotes, with the notable exception of yeasts . In response to DNA damage caused by ionizing radiation or alkylating agents, PARP binds to strand interruptions in DNA and undergoes rapid automodification with synthesis of long branched polymers of highly negatively charged poly(ADP-ribose) . DNA repair occurs after dissociation of modified PARP from DNA strand breaks . Biochemical data with enzyme-depleted extracts and studies of enzyme-deficient mice show that PARP does not participate directly in DNA repair . Possible roles for poly(ADP-ribose) synthesis are discussed.

J Mol Evol, 1995 Oct, 41(4), 407 - 13
Molecular and phylogenetic analysis of PCR-amplified cyclin-dependent kinase (CDK) family sequences from representatives of the earliest available lineages of eukaryotes; Riley DE et al.; Cyclin-dependent kinase (CDK) and cell division control (CDC2) sequences are strongly conserved among eukaryotes and may complement the use of other sequence families in eukaryotic phylogenetic inference . We synthesized degenerate PCR primers to amplify the catalytic region of CDK homologs in representatives of the earliest available lineages of eukaryotes . CDK family sequence-based, maximum-likelihood distance measurements with neighbor-joining, and Fitch-Margoliash least-squares analyses produced unrooted dendrograms that included protists, yeasts, and higher eukaryotes . Bootstrap confidence estimates supported CDK-based phylogenetic groupings among the protists, fungi, and vertebrates although resolution within these groups was often insignificant . However, Trichomonas vaginalis and Giardia lamblia exhibited two of the most divergent CDK-like sequences consistent with rRNA-phylogenetic inference of early divergence of these eukaryotic lineages . In the evolution from unicellular to multicellular organisms, a constellation of amino acid residues aligning with the human, CDK N-terminal beta sheet may have undergone abrupt replacement.

Mycoses, 1995 Sep-Oct, 38(9-10), 421 - 4
Aetiology of onychomycosis in Al Ain, United Arab Emirates; Nsanze H et al.; A 1-year study was conducted on 151 patients with finger- or toenail disease attending a dermatology clinic in Al Ain, United Arab Emirates . Nail scrapings and or clippings were collected and investigated with a KOH preparation for direct microscopy and cultured for fungi . Eighty-nine (59%) patients were females . Most females (62%) presented with fingernail disease while in men the toenails were involved in 76% . Fungi were isolated in 78 (52%) of all cases and the fingernails were more commonly positive than toenails . Of the 78 isolates, 49 (63%) were yeasts, including eight Candida species, dermatophytes accounted for 22 (28%) and seven (9%) were other moulds . Non-dermatophyte moulds constituted 20% of male isolates but only 4% of female isolates . Eighty-one per cent of Candida species were from fingernails, while 87% of all the dermatophytes were from toenails.

Br J Cancer, 1995 Sep, 72(3), 583 - 8
Prostate-specific membrane antigen: evidence for the existence of a second related human gene; Leek J et al.; Prostate-specific membrane antigen (PSM) is a glycoprotein recognised by the prostate-specific monoclonal antibody 7E11-C5, which was raised against the human prostatic carcinoma cell line LNCaP . A cDNA clone for PSM has been described . PSM is of clinical importance for a number of reasons . Radiolabelled antibody is being evaluated both as an imaging agent and as an immunotherapeutic in prostate cancer . Use of the PSM promoter has been advocated for gene therapy applications to drive prostate-specific gene expression . Although PSM is expressed in normal prostate as well as in primary and secondary prostatic carcinoma, different splice variants in malignant tissue afford the prospect of developing reverse transcription-polymerase chain reaction (RT-PCR)-based diagnostic screens for the presence of prostatic carcinoma cells in the circulation . We have undertaken characterisation of the gene for PSM in view of the protein's interesting characteristics . Unexpectedly, we have found that there are other sequences apparently related to PSM in the human genome and that PSM genomic clones map to two separate and distinct loci on human chromosome 11 . Investigation of the function of putative PSM-related genes will be necessary to enable us to define fully the role of PSM itself in the development of prostatic carcinoma and in the clinical management of this malignancy.

Microbiol Rev, 1995 Sep, 59(3), 325 - 44
Protein trafficking in kinetoplastid protozoa; Clayton C et al.; The kinetoplastid protozoa infect hosts ranging from invertebrates to plants and mammals, causing diseases of medical and economic importance . They are the earliest-branching organisms in eucaryotic evolution to have either mitochondria or peroxisome-like microbodies . Investigation of their protein trafficking enables us to identify characteristics that have been conserved throughout eucaryotic evolution and also reveals how far variations, or alternative mechanisms, are possible . Protein trafficking in kinetoplastids is in many respects similar to that in higher eucaryotes, including mammals and yeasts . Differences in signal sequence specificities exist, however, for all subcellular locations so far examined in detail--microbodies, mitochondria, and endoplasmic reticulum--with signals being more degenerate, or shorter, than those of their higher eucaryotic counterparts . Some components of the normal array of trafficking mechanisms may be missing in most (if not all) kinetoplastids: examples are clathrin-coated vesicles, recycling receptors, and mannose 6-phosphate-mediated lysosomal targeting . Other aspects and structures are unique to the kinetoplastids or are as yet unexplained . Some of these peculiarities may eventually prove to be weak points that can be used as targets for chemotherapy; others may turn out to be much more widespread than currently suspected.

Trends Biochem Sci, 1995 Sep, 20(9), 367 - 71
GPI-anchor biosynthesis; Takeda J et al.; Many eukaryotic proteins bind to membranes using a glycosylphosphatidylinositol (GPI) anchor . GPI anchors are essential in yeasts and probably also in protozoa . Although mammalian cells can survive without GPI anchors, their deficiency in haemopoietic cells cause a haemolytic disease, paroxysmal nocturnal haemoglobinuria (PNH) . Here, we discuss recent progress in our understanding of GPI-anchor biosynthesis that could lead to a better understanding of PNH and chemotherapeutic agents to treat protozoal and fungal infections.

Trends Biochem Sci, 1995 Sep, 20(9), 357 - 61
Biochemical and genetic dissection of mitotic chromosome condensation; Hirano T; The molecular mechanisms responsible for mitotic chromosome condensation are unknown . Two independent approaches, biochemical studies in vertebrate cells and genetic analyses in yeasts, have converged recently, leading to the identification of a family of putative ATPases that play a fundamental role in this process . Further characterization of these proteins promises to uncover a highly dynamic aspect of mitotic chromosome architecture.

Biol Pharm Bull, 1995 Aug, 18(8), 1057 - 9
Structure and activity of HYI killer toxin from Hansenula saturnus; Komiyama T et al.; The primary structure of HYI killer toxin produced by Hansenula saturnus was determined in its reduced and pyridylethylated form, as well as the peptides resulting from protease digestion . It was found that the HYI killer toxin consisted of 87 amino acid residues and the molecular weight was calculated to be 9543 Da . It showed 87% homology with HM-1 killer toxin produced by H . mrakii, while there were multiple mutations including one amino acid deletion which, nevertheless, did not alter the strong cytocidal effect on sensitive yeasts.

Clin Orthop, 1995 Aug, (317), 223 - 6
Blastomycosis of the forearm synovium in a child; Albert MC et al.; Blastomyces is a pathogen that may infect healthy and immunocompromised hosts . Frequently entering the lungs, the initial pneumonitis usually is followed by skin and bony involvement secondary to hematogenous spread . Infection of the synovium without bone or skin involvement in the pediatric population has not been reported previously . A soft tissue mass of the forearm in a 9-year-old boy was the initial manifestation of blastomycosis . Radiographic images, ultrasound, and magnetic resonance imaging of the forearm could not distinguish inflammation from sarcoma . Frozen section during open biopsy revealed a granulomatous process and several budding yeasts . There was no evidence of a malignancy . Cultures of the tissue grew Blastomyces dermatitidis . The patient was treated successfully with surgical debridement of the mass and amphotericin B followed by itraconazole, for a total therapeutic course of 18 weeks . Although rarely diagnosed in children, blastomycosis must be included in the differential diagnosis of any soft tissue mass.

J Pediatr, 1995 Aug, 127(2), 322 - 8
Oropharyngeal candidiasis in immunocompromised children: a randomized, multicenter study of orally administered fluconazole suspension versus nystatin . The Multicenter Fluconazole Study Group; Flynn PM et al.; OBJECTIVE: To compare the efficacy, safety, and tolerance of fluconazole suspension versus nystatin in the treatment of oropharyngeal thrush in immunocompromised children . DESIGN: Multicenter, randomized, observer-masked trial . SETTING: Thirty-two centers participated, including hospitals and ambulatory care clinics . PATIENTS: We enrolled 182 immunocompromised infants and children, ages 5 months to 14 years, with signs of oral thrush and presence of yeasts on potassium hydroxide- or gram-stained preparations . Subjects were randomly assigned to receive a single daily dose of fluconazole suspension, 2 to 3 mg/kg per day, or nystatin, 400,000 units four times daily for 14 days; 159 patients, who had culture confirmation of thrush and received at least 7 days of study drug, were evaluated for efficacy; all patients were evaluated for safety . RESULTS: Clinical cure was demonstrated in 91% of the subjects in the fluconazole group and 51% of the subjects in the nystatin group (p < 0.001), and eradication of the organism cultured at entry occurred in 76% and 11% (p < 0.001), respectively . Gastrointestinal conditions developed in six patients who received fluconazole and in three who received nystatin; two fluconazole recipients were subsequently withdrawn from the study . Laboratory abnormalities occurred with equal frequency in both groups . Clinical relapse rates were similar in both groups at 2 weeks (18% and 24% for fluconazole and nystatin, respectively) and 1 month (28% and 27%, respectively) after the completion of study drug . CONCLUSIONS: Fluconazole suspension is more effective than nystatin in the treatment of thrush in immunocompromised children . Both regimens were well tolerated.

Arch Dermatol, 1995 Aug, 131(8), 937 - 42
Terbinafine for the treatment of pedal onychomycosis . A foot closer to the promised land of cured nails?
Shear NH, Gupta AK.
Onychomycosis of the nails includes fungal infection caused by primary nail pathogens that invade the healthy nail plate and secondary invasion in nails with preexisting disease . Onychomycosis may be due to secondary infection from tinea pedis and is the most common nail disease, accounting for roughly 30% of all cutaneous fungal infections . In a survey of 9332 British adults, 16 years of age and older, the prevalence of dermatophyte infection was 2.6% to 2.8% . In subjects 55 years of age or older, the prevalence was 4.7% . In the United States, a similar prevalence has been reported . Onychomycosis can have a significant psychological effect on the subject . In a North American study, the relative composition of isolates from nails was dermatophytes (91%), yeasts (5%), and molds (4%) . Candida is isolated predominantly from fingernails . Uncommonly, there are mixed infections . In other parts of the world, the frequency of these agents may vary.

Biochim Biophys Acta, 1995 Jul 25, 1263(1), 79 - 85
A Chlamydomonas homologue to the 14-3-3 proteins: cDNA and deduced amino acid sequence; Liebich I et al.; We have isolated and sequenced a 1464 bp cDNA from the unicellular green alga Chlamydomonas reinhardtii encoding an acidic polypeptide (259 aa) with considerable homologies to the 14-3-3 proteins of animals, yeasts and higher plants . Like the other members of this highly conserved protein kinase regulatory protein family, the deduced amino acid sequence of the Chlamydomonas 14-3-3 protein includes two putative phosphorylation sites within the N-terminal region (positions 62 and 67) . Furthermore, an EF hand motif characteristic for Ca(2+)-binding sites is located within the C-terminal part of this polypeptide (positions 208-219) . EF hand motifs are also present in the 14-3-3 proteins of some higher plants but not in those of animals and yeasts.

Antonie Van Leeuwenhoek, 1995 Jul, 68(1), 57 - 63
Intraspecific variability and exopolysaccharide production in Aureobasidium pullulans; Yurlova NA et al.; Forty seven strains of the black yeasts, Aureobasidium pullulans and Hormonema dematioides, and the type strain of Hormonema macrosporum were examined using PCR-ribotyping and universally primed PCR with subsequent hybridization . Four groups (populations) were distinguished within A . pullulans with PCR-ribotyping, which largely coincided with UP-PCR/hybridization groups . The UP-PCR technique revealed a greater degree of heterogeneity between the groups studied . Five strains identified as Hormonena dematioides on the basis of physiological and morphological data formed a group recognizable with PCR-ribotyping and UP-PCR/hybridization, which also included H . macrosporum . Aureobasidium pullulans is characterized by the absence of RsaI restriction sites in rDNA amplified with primers 5.8S-R and LR7, while Hormonema species possessed several bands after RsaI digestion . For analysis of distance between populations, PCR-ribotyping with AluI and MspI is sufficient . Strains of A . pullulans produce exopolysaccharides in liquid media with different nitrogen sources, while the strains of Hormonema synthesize minor amounts of polysaccharides in media with peptone . Populations of A . pullulans differ slightly from each other in their optimal, medium-dependent production of polysaccharides.

Trends Biochem Sci, 1995 Jul, 20(7), 261 - 4
Antisense snoRNAs: a family of nucleolar RNAs with long complementarities to rRNA; Bachellerie JP et al.; A growing subset of small nucleolar RNAs (snoRNAs) contains long stretches of sequence complementarity to conserved sequences in mature ribosomal RNA (rRNA) . This article reviews current knowledge about these complementarities and proposes that these antisense snoRNAs might function in pre-rRNA folding, base modification and ribosomal ribonucleoprotein assembly, in some cases acting as RNA chaperones.

J Biol Chem, 1995 Jun 30, 270(26), 15467 - 70
Expression of human RAD52 confers resistance to ionizing radiation in mammalian cells; Park MS; Processing of mutagenic DNA damages by the double strand breaks (DSB) in eukaryotes is most likely achieved by multiple pathways, including homologous recombination . Although RAD52 has been shown to be important for DSB repair in yeasts, its role in DSB repair in mammalian cells has not been demonstrated . This study reports for the first time that the overexpression of human RAD52 confers enhanced resistance to gamma-rays and induces homologous intrachromosomal recombination in cultured monkey cells . Recombination frequency synergistically increased by the combination of overexpression of RAD52 and ionizing radiation . These observations suggest that homologous recombination mediated by RAD52 is involved in double-stranded break repair in mammalian cells.

J Infect Dis, 1995 Jun, 171(6), 1559 - 62
Interactions between human bronchoalveolar macrophages and Blastomyces dermatitidis conidia: demonstration of fungicidal and fungistatic effects; Sugar AM et al.; The effects of human bronchoalveolar macrophages (BAMs) on Blastomyces dermatitidis conidia were investigated . Macrophage monolayers were incubated for 5 days with or without interferon (IFN)-gamma or macrophage colony-stimulating factor (M-CSF) before challenge with conidia for 48 h at 37 degrees C . Hyphal growth (germination) was inhibited 12% by resident BAMs (P < .05) . In contrast, resident BAMs blocked conidial phase transition by 88% (P < .05) . Intermediate forms, aggregates of large cells in clums (not conidia or yeasts by morphologic criteria), appeared after incubation with BAMs and grew into typical yeasts once removed from the BAMs . IFN-gamma and M-CSF inhibited the ability of BAMs to block phase transition but not germination . Human BAMs demonstrated fungicidal and fungistatic activity against B . dermatitidis conidia independent of reactive oxygen intermediates and had greater effects on phase transition than on germination . The phase transition-associated fungicidal and fungistatic activities of BAMs were inhibited by IFN-gamma and M-CSF.

J Clin Microbiol, 1995 Jun, 33(6), 1635 - 7
Relevance of culturing Candida species from intravascular catheters; Khatib R et al.; The significance of isolating Candida species from intravascular catheters was examined retrospectively . Our findings demonstrate that these organisms are encountered among patients with proven invasive candidiasis or as an isolated finding . Febrile patients with yeasts in two or more additional sites may be at risk for invasive cadidiasis and may benefit from treatment.

Clin Infect Dis, 1995 Jun, 20(6), 1535 - 9
Epidemiology of fungal infections: the promise of molecular typing; Pfaller MA; As the incidence of invasive fungal disease-particularly nosocomial candidal infection-has increased significantly over the past two decades, molecular typing has become increasingly important for the development of rational infection-control measures and therapeutic strategies . Numerous molecular methods have been used to subtype Candida species, including restriction endonuclease analysis of genomic DNA, Southern hybridization analysis, pulsed-field gel electrophoresis, and polymerase chain reaction-based approaches . Increasingly, typing techniques are being applied to other fungal organisms as well . Although an ideal epidemiological typing technique applicable to a wide range of fungal pathogens is not yet available, several molecular-typing methods may permit rapid, simple, and sensitive discrimination of specific strains among the most clinically important species of fungi.

Cell, 1995 May 19, 81(4), 611 - 20
par-1, a gene required for establishing polarity in C . elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed; Guo S et al.; The first cleavage of C . elegans is asymmetric, generating daughter cells with different sizes, cytoplasmic components, and fates . Mutations in the par-1 gene disrupt this asymmetry . We report here that par-1 encodes a putative Ser/Thr kinase with similarity to kinases from yeasts and mammals . Two strong alleles have mutations in the kinase domain, suggesting that kinase activity is essential for par-1 function . PAR-1 protein is localized to the posterior periphery of the zygote and is distributed in a polar fashion preceding the asymmetric divisions of the germline lineage . Because PAR-1 distribution in the germline correlates with the distribution of germline-specific P granules, it is possible that PAR-1 functions in germline development as well as in establishing embryonic polarity.

Plant Cell, 1995 May, 7(5), 611 - 21
Post-transcriptional regulation of nitrate reductase by light is abolished by an N-terminal deletion; Nussaume L et al.; Higher plant nitrate reductases (NRs) carry an N-terminal domain whose sequence is not conserved in NRs from other organisms . A gene composed of a full-length tobacco NR cDNA with an internal deletion of 168 bp in the 5' end fused to the cauliflower mosaic virus 35S promoter and appropriate termination signals was constructed and designated as delta NR . An NR-deficient mutant of Nicotiana plumbaginifolia was transformed with this delta NR gene . In transgenic plants expressing this construct, NR activity was restored and normal growth resulted . Apart from a higher thermosensitivity, no appreciable modification of the kinetic parameters of the enzyme was detectable . The post-transcriptional regulation of NR by light was abolished in delta NR transformants . Consequently, deregulated production of glutamine and asparagine was detected in delta NR transformants . The absence of in vitro delta NR activity modulation by ATP suggests the impairment of delta NR phosphorylation and thereby suppression of delta NR post-translational regulation . These data imply that post-transcriptional control of NR expression is important for the flow of the nitrate assimilatory pathway.

Parasite Immunol, 1995 May, 17(5), 253 - 60
Inhibition of neutrophil oxidative metabolism by trichinellosis patient sera . Parasite origin or host induction?
Bruschi F, Carulli G, Azzara A, Minnucci S.
The presence of sera factors able to inhibit both neutrophil chemotaxis and phagocytosis was observed in all patients studied at two months from infection caused by Trichinella britovi and in most of them after one year . Human neutrophils with eosinophils are able to kill T . spiralis newborn larvae in an ADCC system and their major cytotoxic mechanism is oxidative metabolism products . We evaluated the effect of trichinellosis sera on neutrophil oxidative burst to determine if neutrophils are affected by circulating factors during infection . Cells were incubated with sera from trichinellosis patients . Basal or stimulated Superoxide Anion (SA) production and chemiluminescence in response to different stimulation (PMA, f-MLP, opsonized yeasts) of neutrophils incubated with trichinellosis sera were evaluated and compared with those of cells incubated with control sera . The results show that basal SA production was inhibited by 66% of sera and stimulated by 11% . On the contrary f-MLP stimulated production was significantly increased by 22% sera, and inhibited by none . Chemiluminescence in response to f-MLP or PMA was inhibited by 46 and 80% of sera, respectively . These results show that trichinellosis sera can modulate not only SA production but also other steps of the oxidative burst, irrespective of the stimulating agent, so suggesting that different neutrophil activation pathways are affected . Increased IL-2 levels observed in most of the sera did not correlate with the inhibiting capacity of sera . The hypothesis of a parasite origin of the inhibiting factors is discussed in the light of host-parasite relationship.

J Clin Microbiol, 1995 May, 33(5), 1302 - 7
Effect of increasing inoculum sizes of pathogenic filamentous fungi on MICs of antifungal agents by broth microdilution method; Gehrt A et al.; Inoculum size is a critical variable in development of methods for antifungal susceptibility testing for filamentous fungi . In order to investigate the influence of different inoculum sizes on MICs of amphotericin B, 5-fluorocytosine, itraconazole, and miconazole, 32 clinical isolates (8 Aspergillus fumigatus, 8 Aspergillus flavus, 5 Rhizopus arrhizus, 8 Pseudallescheria boydii, and 3 Fusarium solani isolates) were studied by the broth microdilution method . Four inoculum sizes were studied: 1 x 10(2) to 5 x 10(2), 1 x 10(3) to 5 x 10(3), 1 x 10(4) to 5 x 10(4), and 1 x 10(5) to 5 x 10(5) CFU/ml . The National Committee for Clinical Laboratory Standards reference method for antifungal susceptibility testing in yeasts was modified and applied to filamentous fungi . The inoculum was spectrophotometrically adjusted, and all tests were performed in buffered medium (RPMI 1640) at pH 7.0 with incubation at 35 degrees C for 72 h . MICs were read at 24, 48, and 72 h . Amphotericin B showed a minimum effect of inoculum size on MICs for all species with the exception of P . boydii (P < 0.05) . A significant effect of inoculum size on MICs was observed with 5-fluorocytosine, for which there was an increase of more than 10-fold in MICs against all Aspergillus spp . between inoculum concentrations of 10(2) and 10(4) CFU/ml (P < 0.001) . For itraconazole, the results showed a more species-dependent increase of MICs, most strikingly for R . arrhizus and P . boydii . Miconazole, which was tested only with P . boydii, did not demonstrate a significant effect of inoculum size on MICs . In summary, the effect of inoculum size on MICs for filamentous fungi was dependent upon the organism and antifungal compound tested . Thus, among antifungal compounds, itraconazole and 5-fluorocytosine demonstrated significant inoculum effects, while amphotericin B and miconazole showed comparatively minimum inoculum effects against pathogenic filamentous fungi . Moreover, among filamentous fungi, P . boydii and R . arrhizus exhibited the greatest inoculum effect.

J Clin Microbiol, 1995 May, 33(5), 1164 - 8
Detection of Aspergillus species DNA in bronchoalveolar lavage samples by competitive PCR; Bretagne S et al.; A competitive PCR assay involving the use of bronchoalveolar lavage (BAL) samples for the diagnosis of invasive pulmonary aspergillosis (IPA) was developed . For this purpose, a 1-kb mitochondrial DNA fragment of Aspergillus fumigatus was sequenced . The primers used allowed amplification of A . fumigatus, A . flavus, A . terreus, and A . niger DNAs but not DNAs of other fungi and yeasts . BAL samples from 55 consecutively enrolled patients were tested . Three samples were excluded because of failure of correct amplification of the internal competitive control . Of 28 immunocompromised patients, 6 were PCR positive; 3 died of IPA and their BAL cultures yielded A . fumigatus; and 3 were culture negative and did not develop IPA . Of 15 human immunodeficiency virus-positive patients and 9 immunocompetent patients, 5 and 4, respectively, were both PCR positive and culture negative, and none developed aspergillosis . Thus, PCR confirmed IPA in three patients but gave positive results for 25% (12 of 49) of the patients who did not develop aspergillosis . The predictive value of PCR-positive results seems low for patients at risk for aspergillosis . Moreover, the risk of contamination of reaction buffers or biological samples with Aspergillus conidia seems high and has to be weighed in regard to the potential diagnostic benefit of PCR testing as a routine procedure.

Biochim Biophys Acta, 1995 Apr 26, 1229(2), 175 - 80
Primary structure and properties of the inhibitory protein of the mitochondrial ATPase (H(+)-ATP synthase) from potato; Polgreen KE et al.; The primary structure of the inhibitory protein (IF1) of the potato mitochondrial ATPase has been determined by protein sequencing, and its molecular weight determined by electrospray mass spectrometry . Both are consistent with a 56-residue protein of molecular weight 6697 . This protein shows only weak homology with IF1 sequences from mammals and yeasts, and significant deletions are present compared to these sequences . Homology is strongest in the region between residues 22 and 46 (ox heart numbering), where 5 identities and 6 conserved residues are observed across all five IF1 species . In addition, this region shows homology with protein inhibitors from ATPases other than mitochondrial F1 . It is suggested that this region might constitute an ATPase 'inhibitory motif' . Functional studies show that, unlike IF1 from mammals or yeasts, potato IF1 binds only poorly to ox heart F1, and does not show the ability to exist in 2 (alternate) stable conformations.

Genes Dev, 1995 Apr 15, 9(8), 911 - 24
The origin recognition complex has essential functions in transcriptional silencing and chromosomal replication; Fox CA et al.; The role of the origin recognition complex (ORC) was investigated in replication initiation and in silencing . Temperature-sensitive mutations in ORC genes caused defects in replication initiation at chromosomal origins of replication, as measured by two-dimensional (2-D) origin-mapping gels, fork migration analysis, and plasmid replication studies . These data were consistent with ORC functioning as a eukaryotic replication initiator . Some origins displayed greater replication initiation deficiencies in orc mutants than did others, revealing functional differences between origins . Alleles of ORC5 were isolated that were defective for silencing but not replication, indicating that ORC's role in silencing could be separated from its role in replication . In temperature-sensitive orc mutants arrested in mitosis, temperature-shift experiments caused a loss of silencing, indicating both that ORC had functions outside of the S phase of the cell cycle and that ORC was required for the maintenance of the silenced state.

Antimicrob Agents Chemother, 1995 Apr, 39(4), 996 - 7
Treatment of murine pulmonary blastomycosis with SCH 51048, a broad-spectrum triazole antifungal agent; Sugar AM et al.; The in vitro and in vivo activities of a new broad-spectrum triazole derivative, SCH 51048, against Blastomyces dermatitidis were evaluated . As determined by using the new National Committee for Clinical Laboratory Standards proposed standard for susceptibility testing of yeasts, SCH 51048 was the most active of the four agents tested in vitro against 13 strains of B . dermatitidis . In a well-described murine model of acute pulmonary blastomycosis, SCH 51048 was comparable to amphotericin B and at least 30 times more active than itraconazole . On the basis of these experiments, clinical evaluation of SCH 51048 for the use in treatment of human blastomycosis should proceed.

Clin Microbiol Rev, 1995 Apr, 8(2), 161 - 79
Fungal allergens; Horner WE et al.; Airborne fungal spores occur widely and often in far greater concentrations than pollen grains . Immunoglobulin E-specific antigens (allergens) on airborne fungal spores induce type I hypersensitivity (allergic) respiratory reactions in sensitized atopic subjects, causing rhinitis and/or asthma . The prevalence of respiratory allergy to fungi is imprecisely known but is estimated at 20 to 30% of atopic (allergy-predisposed) individuals or up to 6% of the general population . Diagnosis and immunotherapy of allergy to fungi require well-characterized or standardized extracts that contain the relevant allergen(s) of the appropriate fungus . Production of standardized extracts is difficult since fungal extracts are complex mixtures and a variety of fungi are allergenic . Thus, the currently available extracts are largely nonstandardized, even uncharacterized, crude extracts . Recent significant progress in isolating and characterizing relevant fungal allergens is summarized in the present review . Particularly, some allergens from the genera Alternaria, Aspergillus, and Cladosporium are now thoroughly characterized, and allergens from several other genera, including some basidiomycetes, have also been purified . The availability of these extracts will facilitate definitive studies of fungal allergy prevalence and immunotherapy efficacy as well as enhance both the diagnosis and therapy of fungal allergy.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 778 - 80
Efficacy of the triazole D0870 in a murine model of systemic histoplasmosis; Clemons KV et al.; The efficacy of D0870 was studied in a murine model of systemic histoplasmosis and was compared with that of fluconazole . All regimens of D0870 (1 or 10 mg/kg of body weight given daily or 1, 10, or 100 mg/kg given every other day) and 100 mg of fluconazole per kg given daily provided complete protection from lethality . Comparison of the number of viable Histoplasma capsulatum yeasts remaining in the spleen or liver indicated that D0870 was superior to fluconazole in clearing and curing infection . D0870 was 10- to 100-fold more efficacious than fluconazole in the treatment of experimental histoplasmosis.

Diagn Microbiol Infect Dis, 1995 Mar, 21(3), 129 - 33
Comparison of three methods of antifungal susceptibility testing with the proposed NCCLS standard broth macrodilution assay: lack of effect of phenol red . National Committee for Clinical Laboratory Standards; Sugar AM et al.; Three microtiter plate adaptations of the NCCLS proposed standard for antifungal susceptibility testing were evaluated and compared to the NCCLS broth macrodilution method . Thirteen different fungi, including yeasts and moulds were studied . The first microtiter based method was performed exactly as described for the tube dilution assay, with the exception of performance of the assay in 100 microliter in wells of the microtiter plate . The second assay was the same as the first, except for the deletion of phenol red from the RPMI 1640 . The third microtiter assay was based on the reduction of the formazan dye, XTT, after only 24 hours of incubation . All three microtiter methods compared favorably with the macrodilution method, when visually read after either 24 or 48 hours of incubation . Minimum inhibitory concentrations obtained by the XTT assay were usually lower than those obtained by the methods requiring a visual end point determination . Results were reproducible and comparable to those obtained with the NCCLS method . We conclude that microtiter plate adaptation of the NCCLS proposed standard is feasible and the presence of phenol red does not alter the results with the drugs tested . A 24 hour assay using XTT may provide a quicker and more quantitative method of susceptibility testing of fungi . Further investigation of this approach is warranted.

J Infect Dis, 1995 Feb, 171(2), 320 - 7
Macrophages from human immunodeficiency virus-positive persons are defective in host defense against Histoplasma capsulatum; Chaturvedi S et al.; The phagocytic and fungistatic activity of monocyte-derived macrophages from human immunodeficiency virus (HIV)-positive persons against Histoplasma capsulatum yeasts was determined . Macrophages from HIV-positive patients were profoundly deficient in their capacity to recognize and bind H . capsulatum, but ingestion of bound yeasts was normal . The binding of H . capsulatum by patient macrophages tended to decrease with a decrease in CD4+ T lymphocyte counts . Another major defect was that patient macrophages were more permissive for the intracellular growth of H . capsulatum . Macrophages from 22 of 58 patients showed a > or = 2-fold increase in intracellular growth compared with control macrophages . Thus, in addition to defects in cell-mediated immunity caused by a loss of CD4+ T cells, macrophages from HIV-positive patients exhibit intrinsic defects in macrophage function against H . capsulatum that may contribute to the increased susceptibility of HIV-positive patients to disseminated histoplasmosis.

Protein Sci, 1995 Feb, 4(2), 198 - 208
Replacements in a conserved leucine cluster in the hydrophobic heme pocket of cytochrome c; Lo TP et al.; A cluster of highly conserved leucine side chains from residues 9, 68, 85, 94, and 98 is located in the hydrophobic heme pocket of cytochrome c . The contributions of two of these, Leu 85 and Leu 94, have been studied using a protein structure-function-mutagenesis approach to probe their roles in the maintenance of overall structural integrity and electron transfer activity . Structural studies of the L85C, L85F, L85M, and L94S mutant proteins show that, in each case, the overall fold of cytochrome c is retained, but that localized conformational shifts are required to accommodate the introduced side chains . In particular, the side chains of Cys 85 and Phe 85 form energetically favorable interactions with Phe 82, whereas Met 85 takes on a more remote conformation to prevent an unfavorable interaction with the phenyl ring of Phe 82 . In the case of the L94S mutant protein, the new polar group introduced is found to form hydrogen bonds to nearby carbonyl groups . In all proteins with substitutions at Leu 85, the hydrophobic nature of the heme pocket is preserved and no significant decrease in heme reduction potential is observed . Despite earlier predictions that Leu 85 is an important determinant in cytochrome c electron transfer partner complexation, our studies show this is unlikely to be the case because the considerable surface contour perturbations made by substitutions at this residue do not correspondingly translate into significant changes in electron transfer rates . For the L94S mutant protein, the substitution of a polar hydroxyl group directly into the hydrophobic heme pocket has a larger effect on heme reduction potential, but this is mitigated by two factors . First, the side chain of Ser 94 is rotated away from the heme group and, second, the side chain of Leu 98 shifts into a portion of the new space available, partially shielding the heme group . The Leu 94 Ser substitution does not perturb the highly conserved interface formed by the nearly perpendicular packing of the N- and C-terminal helices of cytochrome c, ruling this out as the cause of this mutant protein becoming thermally labile and having a lower functional activity . Our results show these effects are most likely attributable to disruption of the heme pocket region . Much of the ability of cytochrome c to absorb the introduction of mutations at Leu 85 and Leu 94 appears to be a consequence of the conformational flexibility afforded by the leucine cluster in this region as well as the presence of a nearby internal cavity.(ABSTRACT TRUNCATED AT 400 WORDS)

Yeast, 1995 Feb, 11(2), 101 - 10
Perfect state of Rhodomyces dendrorhous (Phaffia rhodozyma); Golubev WI; After mother-daughter cell conjugation, formation of long holobasidia with terminal basidiospores was observed without mycelium production in Rhodomyces dendrorhous (including the type strain of Phaffia rhodozyma) on polyol-containing media . Basidiospores are not forcibly discharged and germinate by budding . A new genus Xanthophyllomyces (Filobasidiaceae, Tremellales) with a species, X . dendrorhous, is proposed for the teleomorphic state of R . dendrorhous.

Antimicrob Agents Chemother, 1995 Feb, 39(2), 314 - 9
Comparative and collaborative evaluation of standardization of antifungal susceptibility testing for filamentous fungi; Espinel-Ingroff A et al.; The purpose of the study was to evaluate the interlaboratory agreement of broth dilution susceptibility methods for five species of conidium-forming (size range, 2 to 7 microns) filamentous fungi . The methods used included both macro- and microdilution methods that were adaptations of the proposed reference method of the National Committee for Clinical Laboratory Standards for yeasts (m27-P) . The MICs of amphotericin B, fluconazole, itraconazole, miconazole, and ketoconazole were determined in six centers by both macro- and microdilution tests for 25 isolates of Aspergillus flavus, Aspergillus fumigatus, Pseudallescheria boydii, Rhizopus arrhizus, and Sporothrix schenckii . All isolates produced clearly detectable growth within 1 to 4 days at 35 degrees C in the RPMI 1640 medium . Colony counts of 0.4 x 10(6) to 3.3 x 10(6) CFU/ml (mean, 1.4 x 10(6) CFU/ml) were demonstrated in 90% of the 148 inoculum preparations . Overall, good intralaboratory agreement was demonstrated with amphotericin B, fluconazole, and ketoconazole MICs (90 to 97%) . The agreement was lower with itraconazole MICs (59 to 79% median) . Interlaboratory reproducibility demonstrated similar results: 90 to 100% agreement with amphotericin B, fluconazole, miconazole, and ketoconazole MICs and 59 to 91% with itraconazole MICs . Among the species tested, the MICs for S . schenckii showed the highest variability . The results of the study imply that it may be possible to develop a reference method for antifungal susceptibility testing of filamentous fungi.

Nucleic Acids Symp Ser, 1995, (34), 119 - 20
Horizontal transmission of group-I ribozymes: viruses as a carrier of the introns; Yamada T et al.; The nucleotide sequences of two self-splicing group I introns found in viruses that infect the eukaryotic green alga Chlorella were compared with those of 19 nuclear group I introns of 11 green algae, 1 red alga, 2 yeasts and 1 protist . The similarity ranging from 69.5% to 89.0% in any comparison strongly suggested a close relatedness of these introns . Since this kind of viruses infect algal cells residing in protists such as Paramecium, they may serve as a mediator to spread group I introns horizontally among organisms of different phylogenetic kingdoms.

Hum Mol Genet, 1995, 4 Spec No, 1791 - 8
Disorders of peroxisome biogenesis; Braverman N et al.; The peroxisome is a ubiquitous, subcellular organelle containing more than 50 matrix enzymes that participate in a diverse array of metabolic pathways . Failure to assemble normal peroxisomes is the cellular hallmark of Zellweger syndrome and other human disorders of peroxisome biogenesis . Identification of the genes required for peroxisome biogenesis is proceeding at a rapid pace helped immeasurably by work in other species, particularly various yeasts . The ultimate goals of this effort are to identify all of these genes and to understand how their protein products interact to produce normal appearing and functioning peroxisomes . Attainment of these goals will lead to a better understanding of the peroxisome biogenesis disorders, their pathophysiology and treatment.

Proteins, 1995 Jan, 21(1), 57 - 67
Modulation of DNA-binding specificity within the nuclear receptor family by substitutions at a single amino acid position; Zilliacus J et al.; Regulation of gene expression involves a large number of transcription factors with unique DNA-binding properties . Many transcription factors belong to families of related proteins that bind to similar but distinct sequences . In this study we have analyzed how amino acid substitutions at a single position in the DNA-binding domain modulate the DNA-binding specificity within the nuclear receptor family of transcription factors . All possible amino acids were introduced at the first position in the DNA recognition helix, and the specificities of the mutants were analyzed using response elements containing all combinations of bases at two variable base pair positions . All mutant proteins were functional in DNA binding, and could be divided into classes of mutants with different response element specificities . By combining functional data with analysis of the structural effects of the mutations by molecular modeling, we could identify both prohibitive steric interactions as well as positive interactions, such as hydrogen bonds, that function as important determinants for specificity . Only the residues found naturally in the glucocorticoid and estrogen receptors, glycine and glutamate, produce unique binding specificities . The specificities of the other mutants overlap with each other somewhat but the substitutions clearly have potential to contribute to diversity within the nuclear receptor family.

Drugs, 1995 Jan, 49(1), 103 - 20
Amorolfine . A review of its pharmacological properties and therapeutic potential in the treatment of onychomycosis and other superficial fungal infections; Haria M et al.; Amorolfine is a structurally unique, topically active antifungal agent, which possesses both fungistatic and fungicidal activity in vitro . Its spectrum of in vitro activity includes dermatophyte, dimorphic, some dematiaceous and filamentous fungi, and some yeasts . In clinical trials, application of amorolfine 5% nail lacquer once or twice weekly for up to 6 months produced mycological and clinical cure in approximately 40 to 55% of patients with mild onychomycosis 3 months after cessation of therapy . Overall cure and improvement was observed in approximately 85 to 90% of patients with superficial dermatomycoses following treatment with amorolfine 0.25% cream for up to 6 weeks . However, few controlled, comparative trials are available for these different mycoses, and only small numbers of patients have been evaluated to date . Both preparations appear to be well tolerated; only minor localised adverse events have been reported in clinical trials . At present, the major potential indication for topical amorolfine appears to be onychomycosis . Within this clinical setting, amorolfine should be reserved for patients with mild infection without nail matrix involvement . Systemic therapy, however, remains essential for patients with severe intractable onychomycosis involving the nail bed . Evidence to date does not clarify whether the use of adjuvant topical amorolfine reduces the need for systemic therapy in patients with severely infected nails, or whether amorolfine is beneficial in individuals unresponsive to other treatment options.

Acta Derm Venereol Suppl (Stockh), 1995, 191, 1 - 47
Pityrosporum ovale in healthy children, infantile seborrhoeic dermatitis and atopic dermatitis; Broberg A; The occurrence of Pityrosporum ovale was studied in healthy children, children with infantile seborrhoeic dermatitis (ISD) and in patients with atopic dermatitis (AD) . Twenty children with ISD and twenty healthy infants were subjected to culture for P . ovale . Positive cultures were found in 18 of 20 infants with ISD, compared with 4 of 20 controls . The same culture medium containing olive oil as one of the lipids was used to evaluate the frequency of positive P . ovale cultures in 60 patients with AD, 40 patients with rhinoconjunctivitis and/or asthma (RA) and 40 children and young adults with no atopic history (HC) . The results of the quantitative cultures from the forehead did not differ between the groups . P . ovale cultures were positive in 0-20% of children aged 0-10 years and in 60-90% of the 11-20-year-old subjects . Positive P . ovale cultures were found in 87% of 138 healthy children aged 2 months to 15 years when cultures were performed on a medium containing whole fat cows' milk as one lipid source . The largest number of colonies was found among children aged 2-23 months and among children older than 9 years . The occurrence of specific IgE antibodies to P . ovale was evaluated with a skin prick test (SPT) and RAST and compared in 3 groups (AD, RA, HC) of patients aged 0-20 years . Specific IgE were found most often in patients with AD . In patients with AD on different parts of the body, 15% had a positive SPT to P . ovale . In another group of patients, aged 14-53 years, with AD localised mainly to the head and neck area, the SPT was positive in 55% of the patients . Sera from 13 patients with positive SPT to P . ovale were further analysed with IgE immunoblotting using both P . ovale and C . albicans antigens . Simultaneous IgE-binding to both these yeasts was found in 5 sera and these were analysed with RAST-inhibition . Cross-reacting IgE antibodies to P . ovale and C . albicans were found in two of these sera . Cross-reacting sera were pooled and used as an IgE probe in crossed radioimmunoelectrophoresis and Tandem-crossed immunoelectrophoresis . Cross-reacting epitopes were suggested to be located in the mannan polysaccharide of C . albicans and in a high molecular weight fraction of P . ovale.(ABSTRACT TRUNCATED AT 400 WORDS)

Clin Investig, 1994 Nov, 72(11), 878 - 82
Disseminated histoplasmosis in a non-immunocompromised host; Harten P et al.; Histoplasma infections in Europe are rare, and acute disseminated histoplasmosis has only been observed in immunocompromised persons . We describe a case of acute disseminated histoplasmosis in a young, nonimmunocompromised European woman . The probable source of infection was Sri Lanka or the Maldives . At presentation she was severely ill with fever, lymphadenopathy, anemia, thrombocytopenia, hepatosplenomegaly, and polyserositis . Histologically, myelofibrosis and osteosclerosis were observed with extramedullary hematopoiesis . Histoplasma capsulatum yeasts were detected in bone marrow trephine biopsy by methenamine silver staining . Treatment with conventional and liposomal amphotericin B and subsequent itraconazole led to rapid and complete recovery.

Mol Gen Genet, 1994 Oct 28, 245(2), 237 - 45
The mitochondrial processing peptidase from potato: a self-processing enzyme encoded by two differentially expressed genes; Emmermann M et al.; Cytochrome c reductase from potato is a bifunctional protein complex located in the inner mitochondrial membrane, which is involved in respiratory electron transport and processing of mitochondrial precursor proteins . The three largest subunits of the complex share the highest degree of sequence identity with the alpha- and beta-subunits of the soluble processing peptidase (MPP) from fungi and mammals . Evidence is provided that another substoichiometric polypeptide of the cytochrome c reductase complex resembles the alpha-subunit of MPP . A cDNA clone corresponding to the second alpha-MPP protein (alpha-II MPP) encodes a polypeptide of 504 amino acids which is 84% identical to alpha-I MPP . The two different alpha-MPP polypeptides have similar sizes on SDS-polyacrylamide gels but can be distinguished with an antibody raised against a decapeptide that is specific for alpha-II MPP . The presequences of both alpha-subunits of MPP are proteolytically removed by the integrated processing enzyme complex indicating that it acts on the targeting signals of its own precursor proteins . Gene-specific oligonucleotides reveal that the genes encoding alpha-subunit I and alpha-subunit II of MPP are differentially expressed in all tissues analysed but the transcript levels do not vary between tissues.

Mol Cell Biol, 1994 Oct, 14(10), 6983 - 95
Tissue-specific expression of the diazepam-binding inhibitor in Drosophila melanogaster: cloning, structure, and localization of the gene; Kolmer M et al.; The diazepam-binding inhibitor (DBI; also called acyl coenzyme A-binding protein or endozepine) is a 10-kDa polypeptide found in organisms ranging from yeasts to mammals . It has been shown that DBI and its processing products are involved in various specific biological processes such as GABAA/benzodiazepine receptor modulation, acyl coenzyme A metabolism, steroidogenesis, and insulin secretion . We have cloned and sequenced the Drosophila melanogaster gene and cDNA encoding DBI . The Drosophila DBI gene encodes a protein of 86 amino acids that shows 51 to 56% identity with previously known DBI proteins . The gene is composed of one noncoding 5' and two coding exons and is localized on the chromosomal map at position 65E . Several transcription initiation sites were detected by RNase protection and primer extension experiments . Computer analysis of the promoter region revealed features typical of housekeeping genes, such as the lack of TATA and CCAAT elements . However, in its low GC content and lack of a CpG island, the region resembles promoters of tissue-specific genes . Northern (RNA) analysis revealed that the expression of the DBI gene occurred from the larval stage onwards throughout the adult stage . In adult flies, DBI mRNA and immunoreactivity were detected in the cardia, part of the Malpighian tubules, the fat body, and gametes of both sexes . Developmentally regulated expression, disappearing during metamorphosis, was detected in the larval and pupal brains . No expression was detected in the adult nervous system . On the basis of the expression of DBI in some but not all tissues with high energy consumption, we propose that in D . melanogaster, DBI is involved in energy metabolism in a manner that depends on the substrate used for energy production.

Mamm Genome, 1994 Oct, 5(10), 623 - 8
Tissue-specific expression of Ran isoforms in the mouse; Coutavas EE et al.; Ran genes encode a family of well-conserve small nuclear GTPases (Ras-related nuclear proteins), whose function is implicated in both normal cell cycle progression and the transport of RNA and proteins between the nucleus and the cytoplasm . Previous studies of Ran proteins have utilized cell-free systems, yeasts, and cultured mammalian cells . We have now characterized patterns of Ran gene expression in the mouse . Serum starvation suppressed Ran gene transcription in mouse 3T3 cells . Ran mRNA reappeared in cells within 3 h after refeeding . A single Ran mRNA species was detected at low levels in most somatic tissues of the adult mouse . In testis, this Ran mRNA was abundant, as were other larger transcripts . Analysis of testis-derived Ran cDNA clones revealed the presence of two transcripts, one specifying an amino acid sequence identical to that of human Ran/TC4 and one specifying an amino acid sequence 94% identical . Northern blotting and reverse transcriptase-PCR assays with oligonucleotide probes and primers specific for each transcript demonstrated that the isoform identical to Ran/TC4 was expressed in both somatic tissues and testis, while the variant form was transcribed only in testis . The existence of tissue-specific Ran isoforms may help to rationalize the diverse roles suggested for Ran by previous biochemical studies.

J Am Acad Dermatol, 1994 Sep, 31(3 Pt 2), S74 - 7
Topical and surgical treatment of onychomycosis; Cohen PR et al.; Dermatophytes, yeasts, and molds can be the causative organisms of fungal nail infections . Alternatives to the systemic management of onychomycosis include topical and surgical treatments . Traditionally, topical agents used as monotherapy for onychomycosis are only able to inhibit the growth of fungal nail infections; clinical and mycologic cures have recently been observed after treatment with some of the newer preparations . In contrast, surgical treatments almost always need to be used in conjunction with either topical or systemic antifungal therapy . An efficacious topical treatment alternative for onychomycosis involves applying antifungal agents concurrently or sequentially with the removal or debridement of the infected nail structures . Alternatively, the application of antifungal lacquer to fingernail and toenail fungal infections may also be an effective topical therapy for the treatment of less severe forms of onychomycosis.

J Am Acad Dermatol, 1994 Sep, 31(3 Pt 2), S68 - 74
Mycology of nail disorders; Midgley G et al.; Cases of onychomycosis represent up to 30% of diagnosed superficial fungal infections and are caused by three groups of fungi: dermatophytes, yeasts, and nondermatophytic molds . The majority of toenail infections are caused by dermatophytes; Trichophyton rubrum is isolated with the greatest frequency . In infections of fingernails, Candida species can be isolated as frequently as the dermatophytes . Of the molds, Scytalidium species can infect both fingernails and toenails, as well as adjacent skin, and represent 3% of the nail infections in a temperate country such as the United Kingdom but a much higher proportion in tropical countries . Other molds such as Scopulariopsis, Acremonium, and Aspergillus species can infect damaged nails . The isolation of a dermatophyte is always considered indicative of infection, but the presence of other molds, which may be aerial contaminants, must be interpreted with care.

Oral Surg Oral Med Oral Pathol, 1994 Sep, 78(3), 306 - 11
Treatment of Candida-infected denture stomatitis with a miconazole lacquer; Konsberg R et al.; The efficacy of a topically administered miconazole denture lacquer was compared with that of a placebo lacquer in the treatment of Candida-infected denture stomatitis . The study was a double-blind, randomized, controlled clinical trial with two parallel treatment groups . The lacquer was applied once on the fitting denture surface . Follow-up examinations took place on days 3, 7, 14, 21, 28, and 35 . On day 14 the effect of the treatment was assessed . Thirty-six patients were included in the statistical analysis . Eighteen received miconazole and 18 received placebo lacquer . Primary efficacy endpoints were the number of colonies cultured from the palatal mucosa and denture surface on day 14 . Thirteen of 16 patients in the miconazole group A showed < 10 colonies on culture medium on day 14 in the specimens from the palatal mucosa as did 5 of 18 patients in the placebo group B (p < 0.05) . Corresponding results for the denture surface were 6 of 17 and 3 of 18, respectively (p < 0.05) . Reapplication of lacquer was considered necessary (> 100 colonies in at least one sampling site within 14 days) in 35% of the patients from group A and in 83% of the patients from group B . The results indicate that a single application of a miconazole denture lacquer considerably reduces the number of Candida yeasts for a substantial period of time.

Mycopathologia, 1994 Sep, 127(3), 189 - 94
Disturbances in the production of interleukin-1 and tumor necrosis factor in disseminated murine sporotrichosis; Carlos IZ et al.; Production of Interleukin-1 (IL-1) and Tumor Necrosis Factor (TNF) by adherent peritoneal cells from BALB/c mice was measured at week 2, 4, 6, 8 and 10 after intravenous inoculation with 10(6) Sporothrix schenckii yeasts . As compared with age-matched controls, IL-1 and TNF production by adherent peritoneal cells from S . schenckii-infected mice was reduced severely at week 4 and 6 of infection and greater than normal at week 8 and 10 . Moreover, between week 4 and 6 of infection there was a depression of delayed type hypersensitivity response to a specific whole soluble antigen, and an increase in fungal multiplication in the livers and spleens of infected mice . Thus, the deficits of cell-mediated immunity in mice with systemic S . schenckii infection may derive, in part, from impaired amplification of the immune response consequent to abnormal generation of IL-1 and TNF.

Mycoses, 1994 Sep-Oct, 37(9-10), 361 - 5
Systemic treatment of skin candidosis: a randomized comparison of terbinafine and ketoconazole; Jung EG et al.; Terbinafine is an antimycotic drug which has a much higher in vitro activity against dermatophytes than against yeasts . To investigate the clinical relevance of these in vitro data, 118 patients with cutaneous candidosis were enrolled in a randomized, double-blind study and allocated to a 4-week treatment with a daily dose of either 250 mg b.i.d . terbinafine or 200 mg once-daily ketoconazole . At the final assessment, 3 weeks after cessation of therapy, mycological cure rates (negative culture and negative microscopy) were 82% in the terbinafine group and 73% in the ketoconazole group . Effective treatment with negative mycology and no or minimal signs or symptoms could be achieved in 65% of those who received terbinafine and in 57% of those randomized to ketoconazole . Five per cent and 7% of the patients taking terbinafine and ketoconazole, respectively, complained about adverse events, which were usually mild and did not lead to discontinuation of treatment . In one patient in the ketoconazole group, abnormal liver enzymes were noted at the final laboratory assessment . The results of this study indicate that terbinafine 500 mg daily can be an alternative to ketoconazole when systemic treatment of skin candidosis is required.

Genetika, 1994 Aug, 30(8), 1022 - 35
{Ambiguity of translation: a eukaryotic version?}; Inge-Vechtomov SG et al.; In this review, the results of studies on genetic control of translation in the yeasts are considered and analyzed . The studies were performed on the system of the SUP35(SUP2) and SUP45(SUP1) genes . Based on data from genetic analysis, the study of protein synthesis in mutants at these genes, sequencing of both genes, deletion analysis of their functions, and the discovery of homologues of these genes in other eukaryotes, it may be assumed that the gene system under study is a specific evolutionarily conservative system for eukaryotes . This system not only controls translational fidelity, but also ensures integration of translation with other cellular processes.

Genetics, 1994 Jul, 137(3), 837 - 43
Gene conversion between unlinked sequences in the germline of mice; Murti JR et al.; Gene conversion between homologous sequences on non-homologous chromosomes (ectopic gene conversion) is remarkably frequent in fungi . It is thought to be a consequence of genome-wide homology scanning required to form synapses between homologous chromosomes . This activity provides a mechanism for concerted evolution of dispersed genes . Technical obstacles associated with mammalian systems have hitherto precluded investigations into ectopic gene conversion in the mammals . Here, we describe a binary transgenic mouse system to detect ectopic gene conversion in mice . Conversion events are visualized by histochemical staining of spermatids, and corroborated by polymerase chain reaction amplification of transgenes in spermatozoa . The results show that conversion between unliked, hemizygous lacZ transgenes is frequent in the male germline, ranging from 0.1 to 0.7% of spermatids . Genomic location may affect the susceptibility to recombination, since the frequency varied between lines . The results suggest that homologous genes can undergo concerted evolution despite being genomically dispersed . However, mechanisms may exist to modulate this activity, enabling the divergence of duplicated genes.

J Eukaryot Microbiol, 1994 Jul-Aug, 41(4), 408 - 14
Trichomonas vaginalis: dominant G2 period and G2 phase arrest in a representative of an early branching eukaryotic lineage; Riley DE et al.; Eukaryotic mitotic cell cycles have been extensively studied in yeasts and vertebrate cells but little is known about cell cycle mechanisms in early branches of the eukaryotic lineage . Trichomonas vaginalis represents one of the earliest branching eukaryotic lineages available for study . In contrast with most yeasts and vertebrate cells, the T . vaginalis G2 period was prolonged, comprising 50 to 58% of the cell population . Hydroxyurea, aphidicolin, and excess thymidine, all of which arrest yeasts and vertebrate cells at the G1/S phase boundary, had no effect on the T . vaginalis cell cycle, probably due to the known absence of synthetic pathways . The anti-microtubule mitotic inhibitors, colchicine and nocodazole, induced G2 phase synchrony . Metronidazole, a therapeutic reagent, also caused G2 phase arrest . These observations suggest that T . vaginalis is similar to yeasts and vertebrate cells in G2 and M phases, but the parasite's G1/S phase transition is distinctive . The results also suggest potentially therapeutic, anti-trichomonad activity of microtubule inhibitors such as nocodazole . The cultured parasite may prove useful as a model for the mitotic cell cycle in the absence of G1/S phase transitional activities universal in yeasts and vertebrate cells.

J Med Chem, 1994 Jun 24, 37(13), 1908 - 17
Derivatives of a novel cyclopeptolide . 1 . Synthesis, antifungal activity, and structure-activity relationships; Emmer G et al.; The synthesis of a series of derivatives of the novel antifungal cyclopeptolide 1, which consists of nine S-amino acids and R-lactic acid, is described . Besides functional group variation of MeAsp4 (esters 2a-d, amides 3a-d, alcohol 4, and its derivatives) and Tyr(Me)9 (demethyl derivative 8, ethers 12a-f, 13, and oxidative degradation of the phenyl group to 14), opening of the lactone by LiOH in THF/H2O allowed manipulation of the hydroxy group of R-Hypr10 in the resulting acyclic peptide 15 . Recyclization of 15 under Mitsunobu conditions followed by deprotection led to the S-Hypr10 analogue 17 of 1 . Cyclic decapeptides 33 and 34 as well as cyclic undecapeptides 35 and 36 were obtained via the corresponding modified linear peptides 23, 24, 27, and 28 by cyclization . Methylation of all secondary amide groups by CH3I and KH/18crown6 gave the permethylated compound 37 . Two of the derivatives (17 and 34) showed superior activities against yeasts in vitro at pH 6.5 as compared to 1, but not at a lower pH (4.5).

Comput Appl Biosci, 1994 Jun, 10(3), 243 - 7
Exploratory analysis of multiple sequence alignments using phylogenies; Golding B; The significance of an alignment between two sequences can be determined using well-known techniques but cannot be easily evaluated with multiple alignments due to the computational complexity . Therefore multiple alignment algorithms may produce an alignment between sequences even when they have little homology with other sequences . A program is presented that makes use of a phylogeny to explore the implications of an alignment . Using the phylogeny, branch lengths are inferred and a search is conducted for regions of unusually rapid or slow rates of change given the observed rates in the rest of the sequence . A very rapid rate of change can be due to either poor homology or due to rapid divergence because of selection . Phylogenies are calculated using either the neighbor joining algorithm of Saitou and Nei (Mol . Biol . Evol., 4, 406-425, 1987) or a phylogeny supplied by the user . The program also permits randomization of subsections of the sequences to determine the significance of the multiple alignment for these individual regions . The combination of these two simple methods permits rapid and interactive exploration of multiple sequence alignments.

J Biol Chem, 1994 May 6, 269(18), 13279 - 88
Purification of a 15-kDa cdk4- and cdk5-binding protein; Azzi L et al.; Yeasts p13suc1/p18CKS and their human homologues, p9CKShs1/p9CKShs2, strongly interact with p34cdc2 and p34cdk2 . While attempting to purify the starfish oocyte p13suc1 homologue, we discovered a 15-kDa protein cross-reactive with anti-p9CKShs2/anti-p13suc1 antibodies . p15cdk-BP-Sepharose binds an anti-PSTAIRE cross-reactive protein of 33 kDa when loaded with starfish oocyte extracts . The p15cdk-BP-bound "PSTAIRE signal" is part of a 250-kDa complex distinct from p34cdc2/cyclin B . p15cdk-BP-Sepharose beads retain a kinase phosphorylating HMG I/Y, P1, and myelin basic protein (among 24 substrates tested) . Major cdc2 kinase substrates are not phosphorylated by the p15cdk-BP-bound kinase . Phosphopeptide maps of P1 phosphorylated by the p15cdk-BP-bound kinase, p34cdc2/cyclin B, p 33cdk5/p25, and casein kinase 2 showed that these kinases phosphorylate P1 on different sites . Phosphopeptide maps of P1 phosphorylated by the p15cdk-BP-bound starfish kinase and p15cdk-BP-bound human p34cdk4/cyclin D are largely coincident . To investigate the nature of the p15cdk-BP-bound kinase, extracts of mammalian tissues and cells were loaded on p9CKShs1- and p15cdk-BP-Sepharose and the bound proteins were analyzed using specific anti-cdk antibodies . cdc2 and cdk2 bind to p9CKShs1-Sepharose, but not to p15cdk-BP . cdk4 and cdk5 bind to p15cdk-BP-Sepharose, but not to p9CKShs1-Sepharose . We conclude that p15cdk-BP specifically binds the cdk4/cyclin D and cdk5 kinases and, along with p13suc1 and p9CKShs, may be part of a larger family of cdk-binding proteins.

J Med Microbiol, 1994 May, 40(5), 350 - 7
Anti-Candida activity of murine bronchoalveolar lavage fluid; Samaranayake LP et al.; Respiratory secretions provide an efficient method for protecting the large surface area of the lower respiratory tract . To determine whether lung secretions contribute to antifungal defences, murine bronchoalveolar lavage fluid (BLF) was tested for anti-candidal activity against 49 oral and vaginal isolates belonging to six different Candida species . The yeasts were incubated in unconcentrated, cell-free lavage fluid from Sprague-Dawley rats and then cultured quantitatively to measure residual viability . Experiments with C . albicans indicated that sensitivity to BLF increased in a time- and dose-dependent manner . This activity was heat-stable (56 degrees C) and consistent, irrespective of whether the BLF was derived from rats inoculated (orally) with candida or the uninoculated controls . Of the Candida spp . examined, C . albicans was the most susceptible followed by C . parapsilosis and C . tropicalis, whereas C . krusei, C . guilliermondii and C . glabrata were highly resistant . However, there were differences in susceptibility to BLF among different isolates within a given species . These results indicate that a heat-stable, soluble factor(s) in murine lavage fluid may suppress candidal colonisation of the lower respiratory tract and contribute to the defence mechanisms of the lungs.

Br J Dermatol, 1994 Apr, 130 Suppl 43, 12 - 4
A comparison of terbinafine (Lamisil) 1% cream given for one week with clotrimazole (Canesten) 1% cream given for four weeks, in the treatment of tinea pedis; Evans EG; Terbinafine (Lamisil) is a synthetic allylamine antifungal which has been shown to be fungicidal against dermatophytes and a number of yeasts and moulds . Topical application of terbinafine 1% cream has been shown to be a highly effective short-duration therapy for dermatophytosis of the skin . The aim of this multicentre, double-blind, parallel-group study was to compare the safety and efficacy of terbinafine 1% cream, applied twice daily for 1 week, with clotrimazole (Canesten) 1% cream, applied twice daily for 4 weeks, in the treatment of tinea pedis . One hundred and seven patients with mycologically proven tinea pedis received 1 week of terbinafine 1% cream, followed by 3 weeks of placebo, and 104 patients received clotrimazole 1% cream for 4 weeks . Mycological cure (negative microscopy and culture) and effective treatment (mycological cure with no or minimal signs and symptoms) were assessed 4 and 6 weeks after commencing therapy . Mycological cure rates were 93.5% for terbinafine and 73.1% for clotrimazole (P = 0.0001) at 4 weeks . Effective treatment rates at 4 weeks were 89.7% for terbinafine and 58.7% for clotrimazole (P = 0.0001), and 89.7% for terbinafine and 73.1% for clotrimazole (P = 0.002) at 6 weeks . The results show that 1 week of terbinafine b.d . is more effective than 4 weeks of clotrimazole b.d., in terms of both mycological cure and effective treatment.

J Clin Invest, 1994 Apr, 93(4), 1422 - 9
Chloroquine induces human macrophage killing of Histoplasma capsulatum by limiting the availability of intracellular iron and is therapeutic in a murine model of histoplasmosis; Newman SL et al.; We investigated the role of intracellular iron on the capacity of Histoplasma capsulatum (Hc) yeasts to multiply within human macrophages (Mphi) . Coculture of Hc-infected Mphi with the iron chelator deferoxamine suppressed the growth of yeasts in a concentration-dependent manner . The effect of deferoxamine was reversed by iron-saturated transferrin (holotransferrin) but not by iron-free transferrin (apotransferrin) . Chloroquine, which prevents release of iron from transferrin by raising endocytic and lysosomal pH, induced human Mphi to kill Hc . The effect of chloroquine was reversed by iron nitriloacetate, an iron compound that is soluble at neutral to alkaline pH, but not by holotransferrin, which releases iron only in an acidic environment . Chloroquine (40-120 mg/kg) given intraperitoneally for 6 d to Hc-infected C57BL/6 mice significantly reduced the growth of Hc in a dose-dependent manner . At 120 mg/kg there was a 17- and 15-fold reduction (P < 0.01) in CFU in spleens and livers, respectively . The therapeutic effect of chloroquine also correlated with the length of treatment . As little as 2 d of chloroquine therapy (120 mg/kg), when started at day 5 after infection, reduced CFU in the spleen by 50% . Treatment with chloroquine for 10 d after a lethal inoculum of Hc protected six of nine mice; all control mice were dead by day 11 (P = 0.009) . This study demonstrates that: (a) iron is of critical importance to the survival and multiplication of Hc yeasts in human Mphi; (b) in vitro, chloroquine induces Mphi killing of Hc yeasts by restricting the availability of intracellular iron; and (c) in vivo, chloroquine significantly reduces the number of organisms in the spleens and livers of Hc-infected mice and can protect mice from a lethal inoculum of Hc yeasts . Thus, chloroquine may be effective in the treatment of active histoplasmosis and also may be useful in preventing relapse of histoplasmosis in patients with acquired immunodeficiency syndromes.

J Immunol, 1994 Apr 1, 152(7), 3491 - 500
Role of CD8+ T cells in host resistance to systemic infection with Histoplasma capsulatum in mice; Deepe GS Jr; Histoplasma capsulatum is a facultative intracellular fungus that spreads lymphohematogenously to involve organ systems rich in mononuclear phagocytes . We explored herein the in vivo effect of anti-CD8 rat mAb on the course of murine disseminated histoplasmosis . Treatment with anti-CD8 mAb caused a selective depletion of CD8+ T cells in naive mice infected with H . capsulatum . The loss of CD8+ T cells was associated with an increased number of H . capsulatum CFU in spleens and livers of mice during the first 3 wk of infection . Transgenic beta 2-microglobulin-deficient mice (beta 2 M-/-) that lack MHC class I Ag and CD8+ T cells or heterozygous littermates were injected with H . capsulatum . At wk 1 of infection, the number of CFU in livers and spleens of beta 2 M-/- mice was similar to that of littermates . At wk 2 and 3, however, the fungal burden in spleens and livers of beta 2 M-/- mice was larger than that of heterozygous littermates . H . capsulatum-immunized mice given anti-CD8 mAb manifested impaired clearance of H . capsulatum upon rechallenge with a large inoculum of yeasts when compared to immunized controls . Histopathologically, the inflammatory response in spleens and livers of CD8+ T cell-depleted mice was similar to that of mice given control Ab . The results indicate that CD8+ T cells are necessary for optimal clearance of the fungus from tissues of mice infected with H . capsulatum.

J Biol Chem, 1994 Apr 1, 269(13), 9946 - 51
Murine cerebellar neurons express a novel gene encoding a protein related to cell cycle control and cell fate determination proteins; Taoka M et al.; We cloned cDNAs of a novel protein (designated V-1) that has been identified from among the developmentally regulated proteins in the rat cerebellum . Protein sequencing analysis (Taoka, M., Yamakuni, T., Song, S.-Y., Yamakawa, Y., Seta, K., Okuyama, T., and Isobe, T . (1992) Eur . J . Biochem . 207, 615-620) and cDNA sequence analysis revealed that the V-1 protein consists of 117 amino acids and contains 2.5 contiguous repeats of the cdc10/SWI6 motif, which was originally found in the products of the cell cycle control genes of yeasts and the cell fate determination genes in Drosophila and Caenorhabditis elegans . In situ hybridization histochemistry revealed that the expression of the V-1 gene is transiently increased in postmigratory granule cells during postnatal rat cerebellar development and thereafter is markedly suppressed, whereas Purkinje cells constitutively express V-1 mRNA . In contrast, cerebellar granule cells of the staggerer mutant mouse continue to express the V-1 gene even when the granule cells of the normal mouse have ceased to express the V-1 gene, suggesting that the expression of the V-1 gene in granule cells is regulated through the interaction with Purkinje cells . On the basis of these results, we postulate that the V-1 protein has a potential role in the differentiation of granule cells.

J Nat Prod, 1994 Apr, 57(4), 518 - 20
Biological activity of some coumarins from Sri Lankan Rutaceae; Gunatilaka AA et al.; Twelve coumarins isolated from plants of the Rutaceae collected in Sri Lanka have been subjected to a mechanism-based anticancer bioassay employing DNA repair-deficient and repair-proficient yeasts . Of these, seselin {10} and xanthyletin {11} were found to be active . Seselin also exhibited moderate cytotoxicity.

Mol Biol Cell, 1994 Apr, 5(4), 465 - 74
Protein import into mitochondria: the requirement for external ATP is precursor-specific whereas intramitochondrial ATP is universally needed for translocation into the matrix; Wachter C et al.; ATP is needed for the import of precursor proteins into mitochondria . However, the role of ATP and its site of action have been unclear . We have now investigated the ATP requirements for protein import into the mitochondrial matrix . These experiments employed an in vitro system that allowed ATP levels to be manipulated both inside and outside the mitochondrial inner membrane . Our results indicate that there are two distinct ATP requirements for mitochondrial protein import . ATP in the matrix is always needed for complete import of precursor proteins into this compartment, even when the precursors are presented to mitochondria in an unfolded conformation . In contrast, the requirement for external ATP is precursor-specific; depletion of external ATP strongly inhibits import of some precursors but has little or no effect with other precursors . A requirement for external ATP can often be overcome by denaturing the precursor with urea . We suggest that external ATP promotes the release of precursors from cytosolic chaperones, whereas matrix ATP drives protein translocation across the inner membrane.

J Virol, 1994 Mar, 68(3), 1728 - 36
Identification and characterization of a protein kinase gene in the Lymantria dispar multinucleocapsid nuclear polyhedrosis virus; Bischoff DS et al.; The Lymantria dispar multinucleocapsid nuclear polyhedrosis virus (LdMNPV) gene encoding vPK has been cloned and sequenced . LdMNPV vPK shows a 24% amino acid identity to the catalytic domains of the eucaryotic protein kinases nPKC from rabbits, HSPKCE from humans, APLPKCB from Aplysia californica, and dPKC98F from Drosophila melanogaster, and homology to several other protein kinases from yeasts, mice, and bovines . The homology suggests that vPK is a serine/threonine protein kinase as defined by Hanks et al . (S.K . Hanks, A.M . Quinn, and T . Hunter, Science 241:42-52, 1988) . Temporal expression studies indicate that vPK is expressed throughout the infection cycle beginning at 4 h postinfection, first as a delayed-early gene and subsequently as a late gene . Sequence analysis and primer extension reactions confirm the presence of distinct early and late transcription initiation regions . Expression of vPK with a rabbit reticulocyte system generated a 31-kDa protein, which is in close agreement with the predicted size of 32 kDa from the amino acid sequence . Phosphorylation activity of in vitro-expressed vPK was demonstrated by using calf thymus histones.

J Biol Chem, 1994 Jan 14, 269(2), 1149 - 53
Specific binding of the peroxisomal protein targeting sequence to glyoxysomal membranes; Wolins NE et al.; It has been demonstrated that the carboxyl-terminal amino acid sequence, serine-lysine-leucine (SKL), is sufficient to direct a polypeptide to peroxisomes in vivo, and that this sequence is functional in plants, animals, and yeasts . Furthermore, many peroxisomal proteins have SKL carboxyl termini, including rat acyl-CoA oxidase . We have synthesized a 125I-peptide with the sequence of the last 12 amino acids of acyl-CoA oxidase, D-Tyr-HKHLKPLQSKL (SKLp), and used it to detect a receptor that recognizes SKL containing proteins targeted to glyoxysomes . SKLp binding to alkali-stripped glyoxysomal membranes was saturable and 80% of the binding could be displaced by 1 microM unlabeled SKLp or 8 micrograms/ml glyoxysomal matrix proteins . Very little specific binding was associated with endoplasmic reticulum or mitochondrial membranes . Specific binding was affected by the ionic composition of the medium; the binding was optimal at pH 6.5 and was inhibited by mono- and divalent cations . Scatchard analysis of SKLp binding to glyoxysomal membranes indicated that there were two binding sites with Kd values of 160 and 1450 nM and abundances of 17 and 43 nmol/mg glyoxysomal membrane protein, respectively . Protease treatment of the alkali-stripped glyoxysomal membranes lowered the number of high affinity sites and destroyed all the low affinity sites . These results demonstrate, for the first time, that there is an integral membrane protein in glyoxysomes that has the characteristics of a receptor for protein import.

Science, 1994 Jan 7, 263(5143), 77 - 81
Catalytic activity of an RNA domain derived from the U6-U4 RNA complex; Yang JH et al.; U6 RNA contains two regions that are essential for proper splicing of nuclear precursor messenger RNA (pre-mRNA) . A comparison of putative secondary structures of the U6-U4 RNA complexes from different phyla revealed a conserved domain that is similar to the catalytic hammerhead RNA motif . Although no catalytic activity was detected in the mammalian U6-U4 RNA complexes, two nucleotide changes in U6 RNA and one in U4 RNA conferred cleavage activity to the complex . Furthermore, the highly conserved domain of the wild-type complex, without the accompanying flanking regions, cleaved an RNA substrate and exhibited other characteristics of the hammerhead ribozyme . The possible involvement of this structure in pre-mRNA splicing is also discussed.

Arch Pathol Lab Med, 1994 Jan, 118(1), 95 - 6
Hyphalike pseudofungus in a lymph node; Teague MD et al.; Mediastinoscopic biopsy specimens of multiple paratracheal lymph nodes in a 66-year-old man with a history of resected pulmonary carcinoma were submitted for histologic evaluation . Structures present in one of these lymph nodes superficially resembled branching septate fungal hyphae . Perls' stain demonstrated that these structures contained iron, and Gomori's methenamine silver stain was negative . The histopathologic features are similar to those of the first and only previous case report of such structures.

Arerugi, 1994 Jan, 43(1), 1 - 8
{Airborne fungi during the last ten years in Sagamihara}; Takatori M et al.; Airborne fungi have been surveyed every week during the 10 years from 1983 to 1992 in Sagamihara . This is a follow up of a 1970 and a 1980 survey at the same sampling station (19.5 m above the ground), using the same gravity potato dextrose agar plate . There were two peak seasons during tsuyu (rainy season; June) and autumn (September to October) . The most predominant fungi found, except for yeasts and non-sporulated fungi, were Cladosporium (2 seasons) and Alternaria (more frequently in tsuyu) . These were followed by Epicoccum (tsuyu), Aureobasidium, Curvularia (summer), Ulocladium (autumn), Penicillium, Arthrinium, Nigrospora (summer), Fusarium, Trichoderma, Pestalotia in decreasing order of their total frequency throughout the 10 years . The most common fungi found during the most years were different from those found during 1970, particularly in the frequency of Aspergillus and Penicillium, but similar to those during 1980 in Sagamihara as well as in most areas of Japan.

Antonie Van Leeuwenhoek, 1994, 65(1), 13 - 20
Investigations of mannan and glucan in the cell wall of Candida boidinii cultivated on methanol; Hristozova T et al.; The polysaccharide components (mannan and glucan) in the cell wall of Candida boidinii M 363 grown on methanol and glucose as control were investigated using electron microscopy, cytochemical and biochemical methods . An ultrastructural rearrangement of the polymers in the cell wall of yeasts cultivated on methanol in comparison to those cultivated on glucose was established . The morphological changes correlate to the quantitative changes in the polysaccharide constituents of the cell wall . The forming and the role of thiosemihydrocarbazide (TSHC)--negative zones in the Candida boidinii cell wall cultivated on methanol media are discussed.

Vet Med (Praha), 1994, 39(6), 329 - 36
{The bioclimate in housing for broilers and its effect on mycotic contamination}; Conkova E; Consequences between climate parameters of stable environment and mould occurrence were monitored during one fattening cycle . Xerophilic and hydrophilic species of moulds were diagnosed in the stable air . Their presence was especially influenced by contaminated feeding mixtures and litter . Mostly the "storage" species of moulds (Penicillium, Aspergillus) but also the "field" species (Fusarium, Alternaria) were isolated from the feeding mixtures . Xerophilic Penicillium species as well as hydrophilic species, e.g . Cladosporium dominated in litter in the beginning of the cycle . Yeasts dominated in the end of the cycle . In examined samples of sedimented dust, the highest frequency of occurrence was in Penicillium germs.

Vet Med (Praha), 1994, 39(6), 321 - 8
{Increased immune response in broilers after administration of natural food supplements}; Kotrbacek V et al.; The trial involved broilers from the 4th to the 56th day of life . The first group P1 (n = 30) received commercial mashes with supplements of 0.5% biomass of freshwater algae (Chlorella vulgaris) . The second group P2 (n = 30) received 0.9% dried cow's colostrum and 0.9% dried brewer's yeasts in addition to the algae (0.2%) . All the ingredients came from fluid-bed drying at temperatures maximally 50 degrees C . Control group K (n = 30) consisted of broilers which received commercial mashes only . At the age of 21, 33 and 56 days, 10 birds of each group were killed, and basic hematological values were determined in the blood samples and phagocytic activity of leucocytes was determined . Samples of the thyme, bursa Fabricii, spleen, ileocecal valve, Meckel's diverticulum, gonads, suprarenal glands and Harder's gland were subjected to histological examination . 22-day chickens of all three groups were vaccinated with Newcastle disease (the vaccine Avipest, Mevak Nitra) and the levels of specific antibodies in blood serum were determined on days 33, 46 and 56 . 0.5% alga supplement to feed ration increased the live weight of experimental broilers at the end of the second week of life only (p < 0.05) . On the 21st and 33rd day of life, phagocytic activity of leucocytes increased significantly (p < 0.01) in these individuals, while in the second month of life development of the intestinal lymphatic tissue and Harder's gland was largely stimulated . Combination of algae with other biological additives did not significantly influence the live weight of broilers, but it enhanced the above-mentioned effect on the phagocytic activity and lymphatic tissue development.(ABSTRACT TRUNCATED AT 250 WORDS)

Mycoses, 1994 Jan-Feb, 37(1-2), 59 - 60
The fungal flora of zoo animals' ears; Kuttin ES et al.; The mycotic flora of the ears of zoo animals was investigated in a large zoological garden in Germany . Malassezia pachydermatis was isolated from the following animals: big ant-eater, brown bear, common wombat, Eurasian badger, Indian elephant, Mangaliza pig and wide-mouthed rhinoceros . Aspergillus and Penicillium species, yeasts and zygomycetes were also isolated from some animals.

Yi Chuan Xue Bao, 1994, 21(3), 193 - 200
{Estimation of evolutionary distances between protein sequences}; Yang Z; Several estimates of the evolutionary distance between two homologous protein sequences were deduced, taking into account of the variation of replacement rate over amino acid sites . A maximum likelihood estimator was also presented with consideration of different probabilities of replacement among amino acids . Suggestions were made as to the application of these distance estimates to real sequence analysis.

Cancer Invest, 1994, 12(4), 384 - 9
Role of upper endoscopy in evaluation of upper gastrointestinal symptoms in patients undergoing bone marrow transplantation; Vishny ML et al.; We reviewed our upper endoscopy (esophagogastroduodenoscopy, EGD) experience in a group of 65 consecutive patients receiving carmustine (BCNU) 600 mg/m2, cisplatin 200 mg/m2, VP-16 2400 mg/m2, and autologous bone marrow transplantation (BMT) for relapsed or refractory non-Hodgkin's lymphoma or Hodgkin's disease . Forty-one patients (33 with chest irradiation) underwent 48 EGDs for the following symptoms: upper gastrointestinal bleeding (melena and/or hematemesis) (12/48); persistent nausea and vomiting (7/48); odynophagia (25/48); and dysphagia (14/48) . All patients who had dysphagia or odynophagia had endoscopic evidence of severe esophagitis, with confluent erosions or ulcerations . Gastrointestinal bleeding, which presented as melena or hematemesis, was caused by severe esophagitis in 11 of 12 patients . Yeasts were detected in 11/42 histological, or cytological specimens and were isolated in 4/26 cultures . No bleeding or infectious complications occurred in any patient as a result of the EGD procedure . We conclude that severe esophagitis documented by EGD is common in lymphoma patients receiving autologous BMT . Use of EGD, however, did not affect the decision to initiate empirical therapy with amphotericin B for persistent fever.

An Med Interna, 1994 Jan, 11(1), 17 - 20
{A substitute treatment for dialysis?}; Battaner E et al.; One of the major problems regarding the administration of amino acids by intravenous feeding is the use of racemic mixtures that are forbidden by the pharmacological regulations; other current problems is the high cost of obtaining pure amino acids . Because of this, our group has been working in the obtention of L-amino acids (assimilables by living organisms) and ketoacids (used by the body as precursors of racemic amino acid mixtures) in a less expensive and simpler way, with the aim of using these products for different pathologies.

Ann Med Interne (Paris), 1994, 145(6), 424 - 8
{African histoplasmosis: clinical and therapeutic aspects, relation to AIDS . Apropos of 4 cases, including a case with HIV-1-HTLV-1 co-infection}; Geffray L et al.; The authors report 4 cases of African histoplasmosis in Zairans . The diagnostic was based ou cutaneous (4 patients), bones (3), lymph-nodes (2), and bowel-localizations (1) and was confirmed by the presence of Histoplasma duboisiis yeasts in cutaneous biopsy (4 patients), in pus of abscess or cutaneous lesions (3) and in stools (1) . Two HIV seronegatives patients had a good outcome with amphotericin B . One patient died without any treatment . One HIV-1 and HTLV-1 seropositive patient had successful short-term outcome with ketoconazole . Seven others cases of association between African histoplasmosis and AIDS are noted in the literature . They suggest the opportunistic nature of this deep mycosis.

Mycoses, 1994, 37 Suppl 1, 97 - 100
{Therapy with ciclopirox lacquer of onychomycoses caused by molds}; Ulbricht H et al.; 60 patients, suffering from an onychomycosis produced by molds were treated for a maximum of 6 months with ciclopirox nail lacquer (8%) . These molds determined by culture were Scopulariopsis brevicaulis (51 x), Aspergillus niger (6 x), Aspergillus fumigatus (2 x) and Hendersonula toruloidea . Anamnesis gave no hints, why molds were the causing fungi of onychomycosis . The achieved mycological cure rate was determined with 90% (culture) respectively 85% (KOH preparation) . The local treatment with ciclopirox nail lacquer was well tolerated by all patients during the entire treatment period of max . 6 months . No side effects occurred . The local therapy with ciclopirox nail lacquer proved to be as effective in the treatment of onychomycosis caused by molds as in the treatment of onychomycosis due to dermatophytes and yeasts.

Mycoses, 1994, 37 Suppl 1, 89 - 96
{Detection of Exophiala dermatitidis (Kano) De Hoog 1977 in septicemia of a child with acute lymphatic leukemia and in patients with cystic fibrosis}; Blaschke-Hellmessen R et al.; In 1992 black yeasts of the species Exophiala dermatitidis were isolated for the first time from patients at the University Clinics in Dresden . Since that time this relatively rarely detected fungus has been frequently cultivated from clinical specimens . Our observations were: Patient with acute lymphatic leukaemia: In a 3 1/2 years old boy E . dermatitidis was isolated from 8 blood cultures during a septicaemic phase . Elimination of the fungus and decreasing of the fever were reached after removing a central venous catheter and treatment with amphotericin B and 5-fluorocytosine for 3 weeks . In this patient E . dermatitidis was assessed to be the cause of the septicaemia setting in via catheter . Patients with cystic fibrosis: In 8 of 51 mycologically surveyed patients E . dermatitidis was frequently - in 2 cases for a long time up to 7 months - isolated from sputum specimens . The occurrence of this fungus was considered to be a colonization with subclinical development . In these patients no fungal invasion or systemic mycosis were seen . The administration of itraconazole for 4 respectively 7 months did not succeed in eliminating E . dermatitidis out of the respiratory tract . It is recommended to include mycological longtime cultures in the surveillance of cystic fibrosis patients for detection of E . dermatitidis.

J Med Vet Mycol, 1994, 32(5), 373 - 80
Exophiala jeanselmei var . lecanii-corni, an aetiologic agent of human phaeohyphomycosis, with report of a case; De Hoog GS et al.; Characters of morphology and nutritional physiology of Exophiala jeanselmei var . lecanii-corni are described . The taxon was proven to be distinct from the var . jeanselmei and from Exophiala dermatitidis by physiology, which is supported by molecular characters . A case of cutaneous phaeohyphomycosis caused by var . lecanii-corni is reported.

Mycoses, 1994, 37 Suppl 2, 77 - 82
{Future directions of antimycotic therapy}; Meunier F; There are presently numerous developments in the field of antifungal armamentarium . Amphotericin B has been considered as the standard treatment since the fifties for most invasive fungal infections . However, emergence of resistant strains and failures despite adequate treatment with amphotericin B are numerous and toxicity as well as side effects are limiting factors in many patients . Alternative modalities to administer amphotericin B using liposomal or lipid formulations are presently evaluated . Tremendous research has been performed in the development of new azoles including ketoconazole, fluconazole and itraconazole . The emergence of resistant strains of yeasts in patients treated with fluconazole requires attention . The problem of possible cross resistance for all azoles should be a matter of further investigation . Itraconazole is a new broad-spectrum antifungal agent . But due to its pharmacokinetic properties monitoring of serum concentrations is recommended in life-threatening situations . New formulations of itraconazole are currently under development and may be extremely useful . The role of azole antifungals for prophylactical administration or therapeutic purposes must be balanced carefully . Studies designed to evaluate adjuvant antifungal treatment based on G-CSF, M-CSF or GM-CSF in combination to antifungals would be useful . Future directions of antifungal treatments should include evaluation of combination of antifungal agents to each other . There is a tremendous need for pivotal clinical studies in the field of antifungal treatments which need international collaboration in order to collect rapidly significant numbers of data . Such comprehensive approaches are initiated by the EORTC Invasive Fungal Infections Cooperative Group, and all European clinicians taking care of mycosis patients are invited to cooperate.

Curr Genet, 1993 Dec, 24(6), 548 - 50
Electrophoretic analysis of the nuclear and organellar genomes in the ultra-small alga Cyanidioschyzon merolae; Maleszka R; Electrophoretic analysis reveals that the nucleus of the ultra-small eukaryotic alga Cyanidioschyzon merolae contains approximately 11.7 x 10(6) base pairs (11.7 Mb) of DNA . This compact genome is fragmented into 15 small chromosomes ranging in size from 410 to 1700 kb . The migratory behaviour of chloroplast DNA is consistent with the presence of a circular plastid genome of about 170 kb . The conformation of mitochondrial DNA resembles that in yeasts and fungi and is predominantly linear and heterogenous in size.

Br J Dermatol, 1993 Dec, 129(6), 696 - 9
Folliculitis in Down's syndrome; Kavanagh GM et al.; Twenty-two male and 20 female adults with Down's syndrome were examined . Ten of the men and two of the women had a follicular rash consistent with Malassezia folliculitis . Oral itraconazole treatment produced a significant improvement in the rash, accompanied by a decrease in the skin Malassezia count . Clinical relapse occurred when therapy was discontinued, and was accompanied by return of the Malassezia yeasts.

Mol Biochem Parasitol, 1993 Dec, 62(2), 243 - 9
Mutational analysis of a signal sequence required for protein secretion in Leishmania major; Tobin JF et al.; In eukaryotes, amino-terminal extensions, signal sequences, mediate the translocation of lysosomal, membrane and secreted proteins into the lumen of the endoplasmic reticulum (ER) . Structure/function studies indicate that eukaryotic signal sequences are composed of 3 distinct domains, a positively charged amino-terminal domain, a central hydrophobic domain, and a polar carboxy-terminal domain . In an attempt to better understand protein trafficking in Leishmania we have constructed strains of Leishmania major that secrete an exogenous protein, interferon-gamma (IFN-gamma), under the control of a mammalian signal sequence . In this report we present a mutational analysis of this signal sequence . Deletion of the entire signal sequence or the hydrophobic core region prevents secretion of IFN-gamma and results in cytoplasmic expression of the protein . Mutations in the amino-terminal domain indicate that a net positive charge is not required for efficient secretion of IFN-gamma . Mutations in the carboxy-terminal domain are more complex and display two phenotypes, either they prevent expression of IFN-gamma or they have no effect on protein secretion . These results indicate that the function of the signal sequence in targeting proteins to the ER in Leishmania is similar to that observed in yeasts and higher eukaryotes and suggests that the Leishmania protein secretory apparatus may also be similar.

Infect Immun, 1993 Nov, 61(11), 4607 - 14
Local immunity in lung-associated lymph nodes in a murine model of pulmonary histoplasmosis; Fojtasek MF et al.; Local immunity against acute pulmonary histoplasmosis was studied in the lung-associated lymph nodes of normal nonimmune mice infected intratracheally with live Histoplasma capsulatum yeasts . The phenotypes and distribution of cells in lung-associated lymph nodes and spleens were determined by flow cytometry . In addition, the immune responsiveness of these cells was evaluated by in vitro blastogenesis . Anti-H . capsulatum antibodies in serum and H . capsulatum antigen in tissue were measured by immunoassays . Cellular immune responses were greater in the lymph nodes than in the spleens . In lymph nodes 7 days after infection, a marked increase in the number of B lymphocytes caused the percentage to rise to 43%, compared with 26% in controls, and it remained elevated throughout the course of infection . A CD3+ cell that did not express CD4 or CD8 increased in number until it constituted 21% of lymph node cells, compared with 5% in controls, by day 14 . The numbers of CD4+ and CD8+ T lymphocytes were modestly increased from days 7 to 35, but their percentages dropped because of the greater numbers of B lymphocytes and CD3+4-8- cells . Macrophages consistently constituted 2 to 3% of lymph node cells during the study . In spleens 7 days after infection, the percentage of macrophages in infected mice rose to 21%, compared with 9% in controls, but the total spleen cell number did not increase until day 14, when all cell subsets were nearly double in number . The in vitro blastogenic response of lymph node cells to H . capsulatum peaked at day 7, but spleen cell response was minimal during the course of infection . Histoplasma-specific serum immunoglobulin G antibodies reached peak levels by day 21 and remained high to the end of the study . In contrast, levels of antigen-specific immunoglobulin M antibodies were very low . These data suggest that antigen-specific immune responses occur in lung-associated lymph nodes and that this draining lymph node response may be an important component in host defense against Histoplasma lung infection.

Trends Genet, 1993 Nov, 9(11), 395 - 9
Thinking about genetic redundancy; Thomas JH; Partial functional redundancy among genes is frequently observed in a wide range of organisms and processes, but the selective value of such redundancy is not immediately apparent . Any fully redundant function should be evolutionarily unstable: unless selection acts to maintain the redundancy it will tend to be lost by mutational drift . I discuss four possible mechanisms by which selection might act to maintain genetic redundancy.

J Fla Med Assoc, 1993 Nov, 80(11), 763 - 5
Anchored proteins . Meeting of basic and clinical research; Adair WL Jr; A novel modification of proteins that make them lipophilic and capable of being tightly associated with membrane has been discovered in a variety of eukaryotic organisms, including man . Known as glycosyl phosphatidylinositol (GPI) anchor, it is profoundly deficient in the hemolytic disease, paroxysmal nocturnal hemoglobinuria (PNH) . The molecular basis for this deficiency is due to the absence of the first enzyme in the biosynthetic pathway for the GPI anchor . Basic research on lower eukaryotes such as trypanosomes and yeasts formed the basis for the clinical discovery.

N Engl J Med, 1993 Oct 21, 329(17), 1231 - 6
Overwhelming pulmonary blastomycosis associated with the adult respiratory distress syndrome; Meyer KC et al.; BACKGROUND AND METHODS . Few cases of overwhelming pulmonary blastomycosis associated with the adult respiratory distress syndrome have been reported . We describe 10 patients with this condition who were treated at one center in Wisconsin . RESULTS . All 10 patients presented with fever, cough, and dyspnea; radiographic evidence of diffuse pulmonary infiltrates; and marked impairment of oxygenation . The mean alveolar-arterial oxygen gradient was 616 mm Hg . Six of the patients had no underlying disease associated with altered immunity, and two had no recent exposure to environmental reservoirs of Blastomyces dermatitidis . In all 10 patients, large numbers of broad-based budding yeasts were seen on microscopical examination of tracheal secretions . All patients were treated with intravenous amphotericin B (0.7 to 1.0 mg per kilogram per day) . Of the five survivors, four received full doses of amphotericin B in the first 24 hours, and four required mechanical ventilatory support for 7 to 151 days . Long-term follow-up of three survivors showed good recovery of pulmonary function . CONCLUSIONS . Overwhelming infection with B . dermatitidis can cause diffuse pneumonitis and the adult respiratory distress syndrome, even in immunocompetent hosts . With prompt diagnosis by microscopical examination of tracheal secretions, intensive therapy with amphotericin B, and ventilatory support, good recovery of pulmonary function is possible.

Gene, 1993 Oct 15, 132(2), 207 - 12
Stable, selectable, integrative DNA transformation in Physarum; Burland TG et al.; The Physarum polycephalum actin promoter, PardC, can drive transient expression of heterologous genes in Physarum amoebae . The hph gene, encoding hygromycin (Hy) phosphotransferase, can confer resistance to Hy on a broad spectrum of organisms . When PardC is translationally fused to hph and transformed into yeasts on high-copy-number vectors, the yeasts become Hy resistant (HyR), showing that PardC-hph is a functional, selectable genetic element . To establish a stable transformation system for Physarum, we electroporated plasmids bearing PardC-hph into Physarum amoebae and then selected for HyR transformants . We show that HyR amoebae arise upon the stable integration of PardC-hph into the nuclear genome in single copy . These results establish a transformation system that can be used to add plasmid-borne genetic information to Physarum.

J Antimicrob Chemother, 1993 Sep, 32(3), 465 - 72
Therapeutic efficacy of a liposomal formulation of amphotericin B (AmBisome) against murine blastomycosis; Clemons KV et al.; The efficacy of a liposomal amphotericin B preparation (AmBisome) and a deoxycholate suspension (Fungizone) were compared in a pulmonary model of murine blastomycosis . Male, four-week-old CD-1 mice were infected intranasally with 19,400 or 27,450 cfu of viable Blastomyces dermatitidis yeasts and intravenous therapy begun 4 days later . Groups of ten mice were given various dosages of Fungizone or AmBisome or were untreated . All treatments were given 3 times per week for 2 weeks . Deaths were recorded over 49 days and the number of viable yeasts in the lungs of survivors quantitated by viable counting . Therapy with 1.0 mg/kg/dose of Fungizone or 1.0, 3.0, 5.0, 7.5 or 20.0 mg/kg/dose of AmBisome were equivalent and prolonged survival over controls and lower dosages of each drug . No acute or chronic toxicities were observed with any regimen . Quantitation of residual numbers of yeasts in the lung showed that 70-100% of mice given 7.5 mg/kg or greater of AmBisome were free of infection and were superior to other regimens . At equivalent 1.0 mg/kg dosages Fungizone was superior to AmBisome . Although AmBisome was three-fold less active than Fungizone on a dosage basis, higher curative doses could be given that were not toxic . These data indicate that AmBisome should be further tested in other models and in clinical trials.

Antimicrob Agents Chemother, 1993 Sep, 37(9), 1962 - 5
Fluconazole resistance in Candida glabrata; Hitchcock CA et al.; We report a case of infection with Candida glabrata in which the organism became resistant to fluconazole and in which pre- and posttreatment isolates were available for comparison . The organism was cross-resistant to ketoconazole and itraconazole, in common with other azole-resistant yeasts . Fluconazole was a potent inhibitor of cytochrome P-450-dependent 14 alpha-sterol demethylase (P-450DM) in lysates of cells from both susceptible and resistant cultures (50% inhibitory concentration, 0.2 microM), indicating that resistance was unrelated to changes in P-450DM . Instead, it appeared to arise from a permeability barrier to fluconazole, since resistant cells were unable to take up radiolabelled drug.

Cas Lek Cesk, 1993 Aug 23, 132(16), 502 - 6
{Trimethoprim in the therapy of symptomatic liver porphyria}; Kalab M et al.; Administration of antimalaria drugs to patients with symptomatic hepatic porphyria (porphyria cutanea tarda) at the First Medical Clinic made it possible due to previous experimental studies, to influence the porphyrim metabolism of yeasts . The effect of trimetoprim was investigated in clinical work in 12 hitherto not treated patients with the manifest form of symptomatic hepatic porphyria . The group comprised 9 men, mean age 56.4 years, and 3 women mean age 43.6 years . During the two-year clinical study dermatological symptoms of the disease became milder or receded . Concurrently there was a significant regression of porphyrinuria . In the whole group porphyrinuria declined to 11% of the original values . The porphyrin content of hepatic tissue declined considerably after two years treatment . In the group as a whole to 40% of the original values . Trimetoprim is another drug which influences porphyrin metabolism . The authors did not detect any undesirable side-effects of trimetoprim treatment . The effect of trimetoprim on clinical and biochemical parameters of the disease is less marked than the effect of chloroquine . This new treatment can be used in patients resistant to chloroquine or in combination with other anti-malaria drugs.

Dig Dis Sci, 1993 Aug, 38(8), 1453 - 8
Association between gastrointestinal tract carriage of Candida, blood group O, and nonsecretion of blood group antigens in patients with peptic ulcer; Burford-Mason AP et al.; Of 112 patients with peptic ulcer disease examined for oral carriage of Candida, 66 (59%) were carriers . Candida carriage was associated with blood group O (P < 0.05) and, independently, with nonsecretion of blood group antigens (P < 0.01) . For each subject, the presence or absence of yeasts was found to be a constant characteristic, and only among patients positive for Candida was blood group O or nonsecretion more frequent than expected in the general population . The quantity of yeasts isolated was significantly greater in patients than in normal subjects (P < 0.002), as was the frequency of carriage in the patient population (59% vs 32%) . This increase was not associated with treatment with H2-receptor antagonists . The results of paired oral and gastroduodenal aspirate cultures suggested that identifying Candida in the oral cavity was a good indicator of the presence of yeasts elsewhere in the gastrointestinal tract . Mechanisms whereby overgrowth of Candida in the upper gastrointestinal tract might contribute to the inflammatory background of peptic ulcer disease are discussed.

No To Hattatsu, 1993 Jul, 25(4), 369 - 73
{An autopsy case of brain candidiasis in premature infant: morphology and intraparenchymal distribution of Candida foci}; Yamaguchi K et al.; An autopsy case of brain candidiasis occurring in a premature infant is presented, and the morphology and intraparenchymal distribution of Candida foci are described in detail with the aid of serial sections of the affected brain . The patient was a boy, who was born after 25 weeks of gestation and died on day 15 . Candida foci were composed of two infectious forms of Candida (yeasts and pseudohyphae) and various inflammatory reactions of the host . They were widely disseminated in the brain parenchyma, leptomeninges and ventricular system . In view of their morphology, they were classified into the acute and chronic inflammatory types . The acute type foci, characterized by microabscess of infiltration of neutrophils, were large and localized predominantly in the cerebral white matter, fiber tracts, central grey matter of the midbrain, reticular formation, floor of fourth ventricle and subependymal germinal layer; most of the acute type foci were found in the watershed zones where the blood supply was considered to be poorer than the other parts of the brain parenchyma . In contrast, the chronic type foci, characterized by nodular proliferation of astrocytes, were small and localized in the grey matter (the cerebral cortex, basal ganglia and brainstem nuclei) and the leptomeninges . This study suggests that Candida infection to the brain may occur by different two kinds of way correlating with the proper vasoarchitecture of brain . In addition, it is recommended to make a close examination of the maternal vagina, placenta and umbilical cord after delivery to detect the risk of Candida infection.

J Gen Microbiol, 1993 Jul, 139 ( Pt 7), 1557 - 64
The production and partial characterization of a monoclonal IgG antibody specific for moulds belonging to the order Mucorales; De Ruiter GA et al.; A monoclonal antibody (mAb) was raised against extracellular polysaccharides from Mucor racemosus after intrasplenic immunization of mice . An indirect ELISA and a dot-blot assay were developed with this mAb . The IgG antibody was found to be very specific for all mould species tested belonging to the order of Mucorales, except species belonging to the genus Mortierella sensu stricto . No cross-reactions were observed with other moulds or yeasts . The immunoreactivity of the polysaccharides of these moulds with this mAb is based on carbohydrate epitopes, in which fucose residues probably play an important role . The mAb may be suited for specific detection of species of the genera Mucor, Rhizopus, Rhizomucor, Thamnidium, Absidia, Syncephalastrum and species belonging to the Mortierella isabellina group in food, and possibly for diagnosis of mucormycosis in humans.

Curr Opin Cell Biol, 1993 Jun, 5(3), 442 - 7
Nuclear organization of pre-mRNA processing; Spector DL; Recent studies have suggested that small nuclear ribonucleoprotein particles (snRNPs), non-snRNP splicing factors, and several heterogeneous nuclear RNP proteins change their organization within the cell in response to transcriptional activity . Several of the RNA substrates with which these factors interact have been shown to localize in tracks that are associated with regions in which splicing factors are concentrated (nuclear speckles) . It is now thought that pre-mRNA splicing may occur within these tracks.

Rev Clin Esp, 1993 Jun, 193(2), 49 - 54
{Candidiasis in an intensive care unit}; Leon Regidor MA et al.; The risk of being infected by candidiasis in an Intensive Care Unit (ICU) is evaluated, using an algorithm which allows the establishment of an early fungicidal treatment . This is a study which includes 34 patients with a mortality of 35% . Yeasts are detected from the second week at ICU, related with the long stay of patients at ICU and the relationship between mortality and age . The first positive sample usually does not indicate disseminate candidiasis (9%) or a positive blood culture (6%) . Population defined as high risk for disseminated candidiasis (HRDC) with a negative blood culture has the worst prognosis (18 cases, 50% mortality rate) . The six cases with HRDC with positive blood culture showed a mortality of 17% . In seven cases there were no HRDC criteria (mortality of 14%) . With said study the existence or not of a HRDC could be determined, establishing the adequate antifungal treatment.

J Biol Chem, 1993 Apr 5, 268(10), 6882 - 5
Cloning of a human gene, PIG-F, a component of glycosylphosphatidylinositol anchor biosynthesis, by a novel expression cloning strategy; Inoue N et al.; The glycosylphosphatidylinositol (GPI)-anchored proteins are widely distributed in eukaryotic cells, from yeasts to mammals . A number of proteins, such as glycosyltransferases, are necessary for GPI anchor biosynthesis . Cloning of genes encoding these proteins is required for analyses of their nature and the biosynthetic pathway . Here we report a new method of expression cloning that is applicable to many mutant rodent and human cells, and its application for cloning a human cDNA termed PIG-F (for Phosphatidyl-Inositol-Glycan class F) using a Thy-1-negative mutant murine thymoma cell line of complementation class F . PIG-F takes a part in the step of transfer of ethanolamine phosphate to the GPI intermediate containing three residues of mannose . This expression cloning strategy is applicable to the identification of not only other genes involved in GPI anchor biosynthesis but also human disease-associated genes using mutant mammalian cell lines.

Pharmacogenetics, 1993 Apr, 3(2), 77 - 85
Hepatic microsomal tolbutamide hydroxylation in Japanese: in vitro evidence for rapid and slow metabolizers; Chen LS et al.; Microsomal hydroxylation of tolbutamide in Japanese livers was studied in vitro to ascertain the enzyme catalysing this reaction . Rates of tolbutamide hydroxylation differed individually 33-fold and 42-fold at 0.1 mM and 2.4 mM tolbutamide concentrations, respectively, and were segregated into two groups, rapid and slow metabolizers . An antibody raised against P450 human-2 (a form of CYP2C9) strongly inhibited the hydroxylation in livers of rapid metabolizers but only weakly inhibited in the slow metabolizer . Kinetic experiments further demonstrated a clear distinction in tolbutamide hydroxylation between two groups; the mean of apparent Km values for tolbutamide was 0.25 mM (n = 3) in the rapid group and 2.58 mM (n = 2) in the slow, respectively . These data suggest that different enzymes are involved in the hydroxylation in both metabolizer groups . Furthermore, CYP2C9 produced by cDNA expression in yeasts, catalysed tolbutamide hydroxylation at rates similar to the rapid metabolizer group at both the 0.1 mM and 2.4 mM concentrations . The apparent Km value of the expressed protein for tolbutamide, 0.26 mM, was similar to that determined for the rapid group of microsomal samples . Clear correlations were observed between the rate of microsomal tolbutamide hydroxylation at 0.1 mM and CYP2C9 protein content or the rate of S-mephenytoin 4'-hydroxylation in human liver . These results indicate that considerable portions of microsomal tolbutamide hydroxylation are catalysed by CYP2C9 or the closely related form in the rapid metabolizers.

J Antibiot (Tokyo), 1993 Apr, 46(4), 631 - 40
Synthesis and antifungal activity of pradimicin derivatives . Modifications of the sugar part; Aburaki S et al.; Modifications at the sugar part of pradimicins were carried out by glycosidations of the aglycones or chemical transformations of natural pradimicins and their antifungal activity was evaluated . Among them, some of the D-xylose-modified derivatives (14, 17, 24) showed activity comparable to that of pradimicin A . The structure-activity relationships obtained through there studies clarified the role of the sugar part in the manifestation of antifungal activity: The 5-O-(6-deoxy-beta-D-sugar) is essential for activity and 2'-epi, 3'-oxo and 4'-deoxy sugar derivatives retain activity against yeasts.

Int J Syst Bacteriol, 1993 Apr, 43(2), 368 - 73
Phylogeny of Metschnikowia species estimated from partial rRNA sequences; Mendonca-Hagler LC et al.; Phylogenetic relationships of species assigned to the genus Metschnikowia were estimated from the extents of divergence among partial sequences of rRNA . The data suggest that the aquatic species (Metschnikowia australis, Metschnikowia bicuspidata, Metschnikowia krissii, and Metschnikowia zobellii) and the terrestrial species (Metschnikowia hawaiiensis, Metschnikowia lunata, Metschnikowia pulcherrima, and Metschnikowia reukaufii) form two groups within the genus . M . lunata and M . hawaiiensis are well separated from other members of the genus, and M . hawaiiensis may be sufficiently divergent that it could be placed in a new genus . Species of the genus Metschnikowia are unique compared with other ascomycetous yeasts because they have a deletion in the large-subunit rRNA sequence that includes nucleotides 434 to 483.

Genitourin Med, 1993 Apr, 69(2), 112 - 4
Management of persistent vulvo vaginal candidosis due to azole-resistant Candida glabrata; White DJ et al.; CASE REPORT--SUBJECTS--Three cases are described of long-standing vaginal candidosis due to Candida glabrata . These had failed to respond to local and systemic antifungals . In each case the infecting strain appeared resistant to a range of azole drugs in vitro . CLINICAL COURSE--Case one--This patient recovered following prolonged treatment with oral itraconazole in combination with oral and vaginal nystatin . Case two . Yeasts were eradicated from this patient following cyclical treatment with oral dydrogesterone; prolonged vaginal treatment with nystatin may have helped . Case three . This patient did not respond to a prolonged course of oral itraconazole in combination with vaginal and oral nystatin, oral medroxyprogesterone or intravaginal boric acid . Eradication of C glabrata was finally achieved by local application of 1% gentian violet . Shortly after eradication of the C glabrata infection, both Case two and Case three developed infections with other Candida species responsive to azole antifungals.

Curr Opin Cell Biol, 1993 Apr, 5(2), 207 - 13
MAP kinase and the activation of quiescent cells; Ruderman JV; Virtually all mitogens lead to the rapid activation of one or more mitogen-activated protein (MAP) kinases . In almost all cases, mitogen-activated surface signaling complexes transmit an essential signal via ras on to a protein kinase cascade that involves the serine/threonine kinase raf . Raf appears to be a MAP kinase kinase kinase, activating MAP kinase kinase which, in turn, activates MAP kinase . Among the targets of MAP kinase are other kinases, nuclear transcription factors and other proteins with roles in cell cycle activation . Both G0-arrested somatic cells and G2-arrested oocytes use many of the same signaling mechanisms to break cell cycle arrest; this is a useful concept in light of newly developed cell-free systems from quiescent oocytes that can be used to study signal transduction in vitro.

Rev Latinoam Microbiol, 1993 Apr-Jun, 35(2), 211 - 6
{Frequency of vaginal trichomoniasis and candidiasis and its relation to the clinical profile}; Sanchez-Vega JT et al.; Eight hundred samples of vaginal swabs were taken from women with active sexual life attending the Cervicouterine Cancer Opportune Detection Service at the Familial Medicine Unit No . 18 of the National Institute of Social Security, to study the incidence and prevalence of vaginal trichomoniasis, the presence of yeasts of the genus Candida and their relationships with clinical features of vulvovaginitis . Identification of parasites was made by means of direct examination with light microscope; 14.39 per cent of candida vulvovaginitis against 3.13 per cent of trichomoniasis was found; in only five cases the two parasites were associated . Leukorrhea and vaginal pruritus were the major symptoms found and the most affected anatomical regions were vaginal walls in trichomoniasis and cervix in candidiasis . Although vaginal trichomoniasis is a worldwide distributed parasitosis, its frequency is very variable, not so vaginal candidiasis, which is more frequent.

J Clin Microbiol, 1993 Mar, 31(3), 572 - 7
The benomyl test as a fundamental diagnostic method for medical mycology; Summerbell RC; The fungicide benomyl has long been known to differentially affect major taxonomic groups of fungi . In the present study 163 species or aggregates of closely similar species of medically important fungi and actinomycetes, as well as species commonly isolated as clinical contaminants, were tested to determine their reactions to three concentrations of benomyl . Fungi of basidiomycetous, endomycetous, and microascaceous affinities were highly resistant, including all common yeasts and Geotrichum, Pseudallescheria, Scedosporium, and Scopulariopsis species . Also resistant were fungi of pleosporalean affinities with poroconidial anamorphs, such as Alternaria, Bipolaris, Curvularia, and Exserohilum species . Most other fungi of ascomycetous affinity were moderately to strongly susceptible . Such fungi included dermatophytes; Coccidioides, Blastomyces, and Histoplasma species; Sporothrix schenckii; medically important aspergilli; and "black yeasts." Benomyl testing aided in the provisional identification of nonsporulating mycelia, including common basidiomycetous isolates obtained as contaminants as well as nonsporulating Aspergillus fumigatus from pulmonary sources.

Br J Biomed Sci, 1993 Mar, 50(1), 27 - 30
Determination of sensitivity to antifungal drugs: evaluation of an API kit; Philpot CM et al.; Evaluation of a commercial kit method for testing sensitivity of yeasts to antifungal drugs was performed employing agar gel dilution as the reference method . For the cidal drugs amphotericin B and flucytosine, results were closely comparable . For the three azole drugs tested, miconazole, econazole and ketoconazole, the discrepant results obtained probably relate to a number of factors affecting the end-point when testing these agents . Although easy to perform, the kit method is relatively expensive and the newer antifungal agents (e.g . fluconazole and itraconazole) are not included . Though showing promise, the kit method has limitations in its present format.

Science, 1993 Feb 26, 259(5099), 1318 - 20
The cloning of PIG-A, a component in the early step of GPI-anchor biosynthesis; Miyata T et al.; The glycosylphosphatidylinositol (GPI) anchor is a membrane attachment structure of many proteins and occurs in a wide variety of eukaryotes from yeasts to mammals . The structure of the core of the GPI anchor is conserved in protozoa and mammals and so is its biosynthetic pathway . A complementary DNA encoding a human protein termed PIG-A (phosphatidylinositol glycan-class A) was cloned . PIG-A was necessary for synthesis of N-acetylglucosaminyl-phosphatidylinositol, the very early intermediate in GPI-anchor biosynthesis.

Cancer Res, 1993 Feb 15, 53(4), 861 - 7
von Hippel-Lindau syndrome: cloning and identification of the plasma membrane Ca(++)-transporting ATPase isoform 2 gene that resides in the von Hippel-Lindau gene region; Latif F et al.; We have isolated and analyzed full-length complementary DNA clones encoded by a 200-kilobase gene encompassing the D3S601 locus that resides in the von Hippel-Lindau (VHL) gene region . The deduced amino acid sequence shows 99% identity with the published sequence of the rat plasma membrane Ca(++)-transporting ATPase isoform 2 complementary DNA, implying that we have cloned and positioned the human plasma membrane Ca(++)-transporting ATPase isoform 2 gene within the VHL critical region . The gene is expressed in VHL target tissues and should be considered a potential candidate gene for the VHL disease.

Cesk Patol, 1993 Feb, 29(1), 32 - 5
{Mycoses of the heart}; Resl M et al.; There is an increasing frequency of heart mycoses . Authors identified 19 such cases between 1972 and 1991, 63% in the last 6 years (frequency was 0.1% in 17.661 autopsies of patients over 7 days of life) . Males prevailed (17:2) . Yeasts of genus Candida dominated, unusual etiology represented Candida parapsilosis, Torulopsis candida and Aspergillus restrictus . Sampling from vegetations or abscesses was a prerequisite for positive autoptic cultivation . Probable source of infection was found in 12 patients and 5 of them had an infected thrombosis of venous catheter . Mycosepsis comprised nearly equal number of endocarditides (mostly left sided) and myocarditides . Spread of endocardial infection into myocardium was rare, most myocarditides resulted from disseminated infection in immunocompromised patients . Endocarditides mainly complicated surgical interventions.

Life Sci, 1993, 52(11), 917 - 26
High-level expression in male germ cells of murine P68 RNA helicase mRNA; Lemaire L et al.; The complete cDNA coding for mouse P68 RNA helicase was cloned and its nucleotide sequence was determined . The sequence is about 95% identical to the human equivalent . Whereas the 5'-untranslated region is less conserved (71%), the 3'-ends of mouse and human mRNAs are nearly identical . Between stop codon and poly(A)-tail both sequences are 97% conserved . At the level of amino acid sequence, the similarity of both, mouse and human, DEAD box family proteins is as high as 98% . In situ hybridizations using cDNA subfragments as probes revealed a testis-selective expression of P68 RNA helicase mRNA . The signal was restricted to late pachytene spermatocytes and haploid spermatids . Northern blot analyses corroborated these results but suggested that expression of related mRNA species occurs in a variety of other tissues.

Hematol Oncol, 1993 Jan-Feb, 11(1), 1 - 5
Protein phosphatase 2A alpha involvement in granulocytic differentiation of HL-60 cells; Sasaki K et al.; Protein phosphatase 2A alpha (PP-2A alpha) in one of the catalytic subunits of PP-2A, which is composed of catalytic and regulatory subunits . In fission yeasts, PP-2A mutants cause mitotic defects and strikingly different cell cycle phenotypes . In rat hepatocellular carcinomas, mRNA expression of PP-2A alpha was increased compared to that of normal hepatocytes . From these studies, PP-2A alpha has been thought to be involved in proliferation, and we expected that PP-2A alpha would decrease in HL-60 cells on dimethyl sulfoxide(DMSO)-induced granulocytic differentiation with decreased growth rate . In this study, we show that PP-2A alpha expression increases in HL-60 cells on DMSO-induced granulocytic differentiation, while PP-1 gamma, which is one of catalytic subunits of protein phosphatase 1, expression did not change significantly . These results suggest that PP-2A alpha may be involved in regulation of differentiation of hematopoietic cells.

Mycoses, 1993 Jan-Feb, 36(1-2), 69 - 73
Incidence of airborne fungi in Isfahan, Iran; Shadzi S et al.; During the one-year period of this study, 288 samples were taken and 954 fungal colonies were isolated from the air of Isfahan, Iran . Among the fungi isolated, Cladosporium, yeasts, Penicillium, Aspergillus and Alternaria were the most frequent isolates respectively . This study showed that environmental factors affect the number and types of airborne fungi, opportunistic fungi, in Isfahan . Sampling location was an important factor: there were more fungal colony counts in the centre and populated locations of the city than in the less populated and rural areas . Regarding collection time, the most colony counts were obtained from samples collected at mid-day and the least in the morning . This study indicated that the incidence of airborne fungi with clinical significance had a direct relationship with the variation of environmental conditions . The results of the present study contribute towards a better understanding of the pattern of occurrence of airborne fungi, and may be useful for allergists, clinicians and epidemiologists.

Mycoses, 1993 Jan-Feb, 36(1-2), 43 - 9
Combination of amorolfine with various antifungal drugs in dermatophytosis; Polak A; The fungistatic activity of ketoconazole, griseofulvin and terbinafine alone and in combination with amorolfine was tested in casitone agar . The fungistatic activity of all three against Trichophyton mentagrophytes is slightly increased by the addition of amorolfine . In yeasts the results are not so clear . The combinations of amorolfine with griseofulvin, terbinafine, itraconazole and fluconazole were always more efficient in a murine model of dermatophytosis than the monotherapy, thus a clear synergism between amorolfine and other antifungals was seen.

Antonie Van Leeuwenhoek, 1993, 64(1), 17 - 26
Nutritional physiology and selective isolation of Exophiala dermatitidis; de Hoog GS et al.; The nutritional physiology of ten strains of Exophiala dermatitidis was investigated . The growth reactions to lactose, citrate, nitrate, nitrite, lysine, creatine and creatinine differ from those found in closely related black yeasts . In addition, it is the only Exophiala species which is able to grow at 40 degrees C . A selective medium containing meso-erythritol as sole carbon source was evaluated . This medium is particularly useful for the isolation of black yeasts from lungs of patients with cystic fibrosis as well as from the environment.

Arch Gynecol Obstet, 1993, 253(3), 157 - 60
Late miscarriage and intraamniotic candidiasis in a woman with a retained intrauterine contraceptive device; Chaim W et al.; PIP: A case report is presented in which a 27-year-old patient, gravida 3, para 2, was admitted at 21 weeks of gestation with uterine contractions . She had had an IUD inserted 3 years before the present pregnancy, and an attempt to remove it when pregnancy was first diagnosed failed . An ultrasound examination at 7 weeks confirmed the gestational age and the presence of an IUD . On admission, ultrasound examination revealed a normal, live fetus with a normal amount of amniotic fluid . The patient was treated with sedation (Meperidine 100 mg and Phenergan 50 mg) and parenteral fluids . Several hours later the cervix was effaced and 4 cm dilated . Under ultrasound guidance, a transabdominal amniocentesis was performed . 20 ml of turbid, greenish amniotic fluid were obtained for Gram stain, white blood cell count, Limulus amebocyte lysate, aerobic, anaerobic and Mycoplasma cultures . Gram stain examination revealed the presence of yeasts . The white blood cell count of amniotic fluid revealed 9000 cells/ml with 73% lymphocytes and 17% polymorphonuclears . The Limulus amebocyte lysate test was negative . The membranes ruptured and a live male fetus weighing 430 g was delivered who died soon afterwards . The placenta was evacuated with the help of suction curettage . The retained IUD was removed with the placenta . Later, Candida was cultured from the amniotic fluid, and from the maternal and fetal sides of the placenta and the fetal skin . The patient was discharged after 5 days of antimycotic treatment was Ketoconazole . The placenta weighed 200 g . The chorionic plate and placental membranes showed histological signs of acute infection with multiple hyphae . The inflammatory infiltrate was composed of granulocytes and fungi . The lungs showed evidence of bronchopneumonia with an inflammatory infiltrate of granulocyte and macrophages . Early diagnosis of the cause of intraamniotic infection accords the best chance of success to antimycotic and other treatments in the prevention of fetal loss .

Int J Syst Bacteriol, 1993 Jan, 43(1), 32 - 5
Validation of the species concept in the genus Dekkera by restriction analysis of genes coding for rRNA; Molina FI et al.; The gene coding for the small-subunit rRNA of 11 type strains belonging to the genus Dekkera and its anamorph, Brettanomyces, was amplified by using the polymerase chain reaction and subjected to digestion with a series of restriction endonucleases . Similarity coefficients were calculated from the number of shared and unique fragments, and a cluster analysis yielded four distinct groups with the following ascosporogenous states: Dekkera anomala, Dekkera bruxellensis, Dekkera custersiana, and Dekkera naardenensis . Results correlate with evidence from isoenzyme electrophoresis and DNA homology analysis . They also confirm previously reported anamorph-teleomorph connections and recently proposed synonymies within the genus.

Curr Genet, 1992 Dec, 22(6), 447 - 54
Sequence analysis of the glyceraldehyde-3-phosphate dehydrogenase genes from the basidiomycetes Schizophyllum commune, Phanerochaete chrysosporium and Agaricus bisporus; Harmsen MC et al.; GPD genes encoding glyceraldehyde-3-phosphate dehydrogenase were isolated from the homobasidiomycetes Schizophyllum commune, Phanerochaete chrysosporium and Agaricus bisporus . All three species contain one transcriptionally active GPD gene, but A . bisporus also contains an inactive GPD gene (tandemly linked to the active gene) . These genes contain 5-9 introns located at conserved positions, differing (except in one case) from intron positions in ascomycetous GPD genes . The predicted amino-acid sequences of the proteins encoded by the three active GPD genes are highly homologous . A comparison with protein sequences from filamentous ascomycetes shows a clear distinction, whereas the GPD genes from ascomycetous yeasts are quite distinct from both the filamentous ascomycetes and basidiomycetes . Promoter regions of ascomycetous GPD genes do not correspond to those of the GPD genes of basidiomycetes which may (partly) explain poor expression in basidiomycetes of introduced genes driven by an ascomycete GPD promoter.

Nihon Kyobu Shikkan Gakkai Zasshi, 1992 Dec, 30 Suppl, 198 - 206
{Mechanisms of clearance of foreign bodies by macrophages}; Suga M et al.; Macrophages have specific functions related to the sites where they are found . Pulmonary alveolar macrophages (PAM) may play an important role the clearance of invading and degenerative substances from the alveolar spaces . To clarify the mechanisms of the clearance by macrophages, we studied the role of scavenger receptors (SR) and Fc receptors (FcR) of PAM . 1) SR: Expression and localization of SR in human PAM . Type I and II SRs were expressed in human PAM, and the two SRs were confirmed in the same cells by double staining method using polyclonal antibodies to the SRs . These SRs were localized on the cell membrane and the endosome in part . The expression of SR in PAM from normal nonsmokers, smokers and patients with pneumoconiosis was studied, but there were no differences among these groups . Kinetics of SR and acetylated low density lipoprotein (acLDL) in PAM . SR recognized acLDL, and SR and acLDL-Au conjugates were concentrated in a coated pit 2 min after incubation . The conjugates were then internalized in the endosome 5 min after incubation . A few acLDL-Au conjugates were present in the endosome near the nucleus, but most of them were found in the lysosome 10 min after incubation . SR was found in the Golgi apparatus 15 min after incubation . These results suggest that SR in PAM may play an important role in the removal of alveolar surfactant . 2) FcR: Effects of FcR mediated phagocytosis on O2-release and phagosome-lysosome fusion in the phagosome of PAM.(ABSTRACT TRUNCATED AT 250 WORDS)

Ukr Biokhim Zh, 1992 Nov-Dec, 64(6), 91 - 4
{Thiochrome inhibition of alcohol dehydrogenase}; Petrov SA; It is shown that thiochrome inhibits alcohol dehydrogenase . Thiochrome is able to be bound with alcohol dehydrogenase more quickly than other thiamine metabolites . This process is specific and has common features with the process of NAD binding by this enzyme . The inhibition of alcohol dehydrogenase by thiochrome is concurrent to NAD . The constant of alcohol dehydrogenase inhibition by thiochrome is 3.9 x 10(-5) M.

Trends Biotechnol, 1992 Nov, 10(11), 388 - 91
Malaria transmission-blocking vaccines; Kaslow DC et al.; Antibodies to surface proteins of the sexual stages of Plasmodium falciparum block completely the transmission of these malaria parasites . Transmission-blocking vaccines therefore represent a powerful and novel approach to controlling the spread of this lethal disease.

Microbiologica, 1992 Oct, 15(4), 367 - 90
Extracellular phenoloxidase activity of micromycetes from various taxonomic groups; Guiraud P et al.; The ability of Micromycetes strains to produce extracellular phenoloxidases was examined on solid malt agar medium using ten different reagents . We established a POx index summarizing the global activity given by the ten reagents used . The results indicated a wide variability depending on the taxonomic groups, the genera and the species . Some groups were relatively homogeneous, either no and low producers of phenoloxidases (Yeasts, Zygomycetes, genera Aspergillus and Penicillium) or medium and high producers of phenoloxidases (Basidiomycetes, Coelomycetes, Tuberculariales and Dematiaceae), while other groups were very heterogeneous (Ascomycetes, Mucedinaceae) . The POx index was significantly higher for strains recently isolated than for strains kept in the fungi collection for a long time.

Sci Am, 1992 Oct, 267(4), 68 - 74B
Histones as regulators of genes; Grunstein M; Histones were once dismissed as little more than packing material for nuclear DNA . In fact, these proteins can both repress and facilitate activation of many genes.

Gene, 1992 Oct 1, 119(2), 175 - 82
Cloning, sequence and expression of a beta-tubulin-encoding gene in the homobasidiomycete Schizophyllum commune; Russo P et al.; The beta-tubulin (beta Tub)-encoding gene (tub-2) of Schizophyllum commune is the first tubulin gene isolated, cloned and sequenced from higher filamentous fungi (homobasidiomycetes) . The S . commune tub-2 gene is organized into nine exons and eight introns . The introns vary from 48 to 107 nt in length, and are distributed throughout the gene . The tub-2 exons code for a protein of 445 amino acids (aa), which shows great homology with beta Tubs of filamentous ascomycetes, plants, and animals, but less homology with yeasts . The codon usage of tub-2 from S . commune is biased, as it is in most beta Tub-encoding genes of filamentous fungi . The S . commune beta Tub shows a conserved aa sequence in the C-terminal domain, which is suggested to interact with microtubule-associated proteins in animals . In contrast, the S . commune beta Tub deviates from most known beta Tubs by having a Cys165 residue, which might be significant for the insensitivity of S . commune haploid strains to the antimicrotubule drug, benomyl . In tub-2 of different haploid strains, sequence polymorphisms occur in the 5' and 3' flanking regions . The expression of tub-2 is high in young mycelium, which has a high number of extending apical cells, but decreases with the aging of the mycelium . No significant difference in the hybridization signal intensity for the tub-2 transcripts was recorded either during intercellular nuclear migration at early mating, or in mycelia with a mutation in the B mating-type gene.(ABSTRACT TRUNCATED AT 250 WORDS)

Nature, 1992 Sep 17, 359(6392), 244 - 6
A vertebrate actin-related protein is a component of a multisubunit complex involved in microtubule-based vesicle motility; Lees-Miller JP et al.; Actin is a cytoskeletal protein which is highly conserved across eukaryotic phyla . Actin filaments, in association with a family of myosin motor proteins, are required for cellular motile processes as diverse as vesicle transport, cell locomotion and cytokinesis . Many organisms have several closely related actin isoforms . In addition to conventional actins, yeasts contain actin-related proteins that are essential for viability . We show here that vertebrates also contain an actin-related protein (actin-RPV) . Actin-RPV is a major component of the dynactin complex, an activator of dynein-driven vesicle movement, indicating that unlike conventional actins which work in conjunction with myosin motors, actin-RPV may be involved in cytoplasmic movements via a microtubule-based system.

Dev Biol, 1992 Sep, 153(1), 1 - 15
Control of the cell cycle; Jacobs T; Cell division is arguably the most fundamental developmental process for single-celled and multicellular organisms alike . The pathway from one cell division to the next is known as the cell cycle . A conserved biochemical regulatory network controls progress along this pathway in plants, animals, and yeasts . This review is intended to serve as a primer on the current state of the eukaryotic cell cycle regulatory model, an introduction to the special roles of cell division and its control in plant development, and a review of recent progress in applying the universal mitotic control paradigm to higher plant systems.

Clin Exp Dermatol, 1992 Sep, 17 Suppl 1, 56 - 60
Double-blind comparison of amorolfine and bifonazole in the treatment of dermatomycoses; Nolting S et al.; A total of 232 patients with mycoses of skin folds, body, or feet were entered into a double-blind, parallel group-study . Therapy with 0.125, 0.25, 0.5% amorolfine cream or 1% bifonazole cream was randomly allocated to patients . The cream was applied once daily for 4 weeks on average . At screening, in 208 patients evaluated for efficacy, a total of 225 fungi were isolated: T . rubrum (77), T . mentagrophytes (65), other dermatophytes (15), C . albicans (34), other yeasts (26) and moulds (8) . One to three weeks after ending therapy, the percentage of patients with negative cultures were as follows: 87.3, 91.7, 90.7 and 92.2% in the amorolfine cream 0.125%, 0.25%, 0.5% and bifonazole cream 1% groups respectively . The differences were not statistically significant . Six out of 223 patients evaluated for safety had local adverse events: one (1.7%), two (3.6%) and three (5.4%) in the amorolfine cream 0.125%, 0.25% and bifonazole cream 1% groups respectively . The most common local adverse events were burning and increased itching, erythema or weeping . A once-daily application of amorolfine cream can be recommended for the treatment of dermatomycoses on the basis of the results from this study . However, a further and similar study with a larger number of patients was required to select the concentration of amorolfine cream for therapeutic use.

Clin Exp Dermatol, 1992 Sep, 17 Suppl 1, 50 - 5
Dose-finding study of amorolfine cream (0.125%, 0.25% and 0.5%) in the treatment of dermatomycoses; del Palacio A et al.; A total of 725 patients with mycosis of the skin folds, large areas of the body or feet were entered into this double-blind, dose-finding study . Treatment with 0.125, 0.25 or 0.5% amorolfine cream was randomly allocated to patients . The cream was applied once daily for 4 weeks on average . At screening, in 527 patients evaluated for efficacy, a total of 533 pathogens were isolated: T . rubrum (322), T . mentagrophytes (84), E . floccosum (45), M . canis (42), other dermatophytes (14), C . albicans (24) and other yeasts (2) . One week after the end of treatment, the culture was negative in 80.5, 81.3 and 84.8% of patients treated with 0.125, 0.25 or 0.5% amorolfine cream, respectively . The differences were not statistically significant . Forty-four out of 714 patients evaluated for safety had local adverse events: 14 (5.8%), 13 (5.5%) and 17 (7.1%) in the amorolfine cream 0.125, 0.25 and 0.5% groups, respectively . Due to local adverse events, six patients (2.5%) in the 0.125% group, six patients (2.6%) in the 0.25% group and seven patients (2.9%) in the 0.5% group discontinued the trial treatment . The most common adverse events were burning, itching, erythema and scaling . No systemic adverse events were reported.

Clin Exp Dermatol, 1992 Sep, 17 Suppl 1, 44 - 9
Comparative efficacy and safety of amorolfine nail lacquer 5% in onychomycosis, once-weekly versus twice-weekly; Reinel D et al.; Amorolfine is a new topical antifungal of the phenylpropyl morpholine class which is highly active both in vitro and in vivo against yeasts, dermatophytes and moulds responsible for superficial fungal infections . Human pharmacological studies have established that amorolfine has a persistent antifungal effect in the nail bed and in the skin without being systemically absorbed . This has been confirmed by clinical work showing that amorolfine is effective in treating dermatomycoses and onychomycoses when administered as cream or nail lacquer . It is ineffective when given orally for systemic mycoses or bacterial infections in animals . In earlier studies a 5% concentration of amorolfine nail lacquer was found to produce a better cure rate in onychomycosis than a lower concentration of 2% . From data available on the penetration of amorolfine and on the persistence of mycologically relevant tissue concentrations, it appeared likely that once- or twice-weekly application of nail lacquer should suffice to produce a satisfactory therapeutic effect in onychomycosis . The aim of this investigation was to assess the efficacy and tolerability of 5% amorolfine nail lacquer given once versus twice weekly to patients with onychomycosis of finger nails and toe nails.

Plant Mol Biol, 1992 Aug, 19(5), 867 - 72
Isolation and characterization of a cDNA clone encoding the TATA box-binding protein (TFIID) from wheat; Kawata T et al.; We isolated a complementary DNA (cDNA) encoding the TATA-binding factor 'TFIID' from a wheat seedling cDNA library . The wheat TFIID transcript of 1.2 kb poly(A)+ RNA was expressed at a low level early in germination, but gradually increased as the seedlings developed . In vitro binding experiments showed that the bacterially expressed wheat TFIID protein could specifically bind to the TATA boxes of the cauliflower mosaic virus (CaMV) 35S, wheat histone H3 and adenovirus major late genes with different affinity . A comparison with Arabidopsis TFIID showed the presence of a plant-specific region consisting of 13 amino acids at the divergent amino terminus and a conserved region (182 amino acids) at the carboxy terminus longer than that observed in yeasts (180 amino acids) and animals (181 amino acids).

Minerva Med, 1992 Jul-Aug, 83(7-8), 475 - 8
{Autoimmune polyendocrinopathy and chronic mucocutaneous candidiasis}; Proto G et al.; A chronic mucocutaneous infection due to yeasts to the Candida genus can be part of the autoimmune polyglandular syndrome, described as APECS, APECED, SPA type I, CES . The authors describe the case of a young patient affected by mucocutaneous candidiasis, hypoparathyroidism and adrenal insufficiency . Chronic mucocutaneous candidiasis was diagnosed first in early childhood and was followed by the development of hypoparathyroidism (4 yrs) and adrenal insufficiency (5 yrs) . The initial lesion of this syndrome is still unknown as the reason of the marked susceptibility to mucocutaneous candidiasis without systemic candidiasis . The treatment of mucocutaneous candidiasis has been improved by the introduction of orally active antifungal drugs as Ketoconazole, first, and fluconazole now.

Genetics, 1992 Jun, 131(2), 307 - 19
Mating type switching in the tetrapolar basidiomycete Agrocybe aegerita; Labarere J et al.; The study of fruiting in the basidiomycete Agrocybe aegerita has shown that some haploid homokaryotic strains can spontaneously switch their mating specificities at the two unlinked A and B mating type factors . This event causes the dikaryotisation of primary homokaryons without plasmogamy and leads to the differentiation of sporulating fruit-bodies (pseudo-homokaryotic fruiting) . For each mating type factor, the genetic analyses have revealed that: (1) parental and switched mating types segregate meiotically as Mendelian markers, (2) a total of six switched mating type factors (two parental and four nonparental) were obtained from a wild strain, (3) most of the nonparental factors have specificities differing from those of a large series of wild factors, (4) strains with the same expressed mating type can generate different specificities, (5) switching is always restricted to the same mating type in a homokaryon, (6) nonparental types can switch again, and (7) meiosis fixes the specificities to which switching can occur . This suggests, for the first time in filamentous fungi, the existence of a mechanism analogous to the mating type switching in yeasts . We hypothese that both A and B mating type regions in A . aegerita are constituted of three loci, one specialized in expression and two other carrying silent information . Mating type switching in homokaryotic strains would occur by copy transposition of silent A and B information into the expression loci . Moreover, we propose that during meiosis the silent loci are substituted by copies of the expressed loci.

Arch Biochem Biophys, 1992 Jun, 295(2), 410 - 20
Isopentenoid synthesis in embryonic Drosophila cells: prenylated protein profile and prenyl group usage; Havel CM et al.; It has been established that vertebrates and yeasts modified a unique subset of polypeptides with farnesyl and geranylgeranyl residues . This observation has been extended to Drosophila Kc cells . {3H}Mevalonate was incorporated into 54 Kc cell peptides (18-92 kDa) . As reported for mammalian cells, most of the labeled peptides had molecular weights between 21 and 27 kDa . C18 radio-HPLC tryptic digest profiles for delipidized, {3H}mevalonate-labeled (a) insect (Drosophila and Spodoptera frugiperda) and mammalian (Chinese hamster ovary met 18-2b) cells, (b) Kc cell nuclear lamin, and (c) a 23.5-kDa purified Kc cell GTP-binding protein were compared and analyzed . {35S}Cysteine-labeled Kc cells yielded a tryptic digest radio-HPLC profile which was congruent with that for {3H}mevalonate-labeled cells . A significant fraction (30-33%) of the doubly labeled tryptic peptides were eluted with greater than or equal to 93% acetonitrile . Kc cell nuclear lamin tryptic digests yielded a single 3H-labeled product which migrated as S-farnesylcysteine . The Kc cell 23.5-kDa GTP-binding protein's 3H-labeled oligopeptide(s)/amino acid(s) was geranylgeranylated and its tryptic digest profile was representative of prenylated proteins whose oligopeptides eluted with greater than or equal to 93% acetonitrile . Moreover, the 3H-labeled oligopeptide/amino acid profiles plus prenyl group patterns for {3H}mevalonate-labeled Kc and mammalian cell total extracts were similar . Collectively, these observations supported a prenylated protein spectrum and prenyl group usage as highly conserved eukaryotic cellular characteristics.

Int J Food Microbiol, 1992 Jun, 16(2), 123 - 30
Fungi causing thread mould spoilage of vacuum packaged Cheddar cheese during maturation; Hocking AD et al.; Thread mould is a defect which occurs sporadically in maturing vacuum packaged Cheddar cheese, caused by the growth of fungi in folds and wrinkles of the plastic film in which the cheese is packaged . Fungi were isolated and identified from 110 Cheddar cheese blocks exhibiting typical thread mould defects . The major causative species were found to be Cladosporium cladosporioides, Penicillium commune, C . herbarum, P . glabrum and a Phoma species . Yeasts were also frequently isolated from the cheese, the majority belonging to the genus Candida . Fungal species which can cause thread mould defects were also found in the cheese factory environment, on cheesemaking equipment, in air, and in curd and whey, providing a wide range of potential sources of contamination.

Ann Hematol, 1992 May, 64(5), 240 - 4
Chronic systemic candidiasis in acute leukemia; Blade J et al.; In the past few years a new syndrome of invasive Candida infection, the so-called hepatosplenic or chronic systemic candidiasis (CSC), has been recognized with increasing frequency in neutropenic patients . From January 1985 to December 1990, ten of 305 acute leukemia (AL) patients treated at our institution were diagnosed as having CSC . In contrast, during the same period this type of Candida infection was not observed in any patient with hematological diseases other than AL treated in our center, including 277 patients who underwent bone marrow transplantation . All patients with CSC had fever and hepatomegaly, and five complained of abdominal pain . Seven patients had neutrophilic leukocytosis and six an increased serum alkaline phosphatase activity . Abdominal computed tomography and ultrasound study showed typical lesions in eight and seven patients, respectively . In four patients a laparoscopy-guided needle liver biopsy displayed yellowish nodules on the liver surface, and the histologic study revealed large granulomas with yeasts and pseudohyphae . All patients were given amphotericin B (mean: 4.6 g, range: 1-12.5 g) and 5-fluorocytosine, and five received fluconazole . No patient died as a direct consequence of CSC and in six the infection resolved . Finally, once controlled, the infectious complication did not preclude subsequent intensive antileukemic therapy, including bone marrow transplantation.

J Prosthet Dent, 1992 May, 67(5), 637 - 44
Management for the highly caries-susceptible patient; Pardo GI et al.; Despite dramatic improvement in caries treatment during the past 30 years, a substantial number of patients remain highly susceptible and do not respond to conventional treatment . It is possible, using simple chairside caries-susceptibility tests, to identify the etiologic factors responsible for the disease and to design a rational approach to treatment that addresses the specific needs of the patient.

Antonie Van Leeuwenhoek, 1992 May, 61(4), 277 - 84
Botryozyma nematodophila gen . nov., spec . nov . (Candidaceae); Smith MT et al.; The new genus Botryozyma with a single species, B . nematodophila is proposed for two isolates from nematodes (Panagrellus zymosiphilus) occurring in grapes with sour-rot . The new genus has typical ascomycetous characteristics and, being unable to produce ascospores, is placed in the family Candidaceae.

Mycoses, 1992 May-Jun, 35(5-6), 121 - 9
Chemiluminescence of polymorphonuclear granulocytes in the presence of selected Candida species; Borg-von Zepelin M et al.; We monitored the respiratory burst of polymorphonuclear granulocytes (PMN) during infection in vitro with 25 strains of six Candida species with different extracellular proteolytic activity by chemiluminescence (CL) . The CL-response of PMNs to viable Candida blastoconidia was compared to the response to ethanol-fixed cells of the same strain . Ethanol-fixed blastoconidia of all tested fungal strains uniformly increased the CL-response on contact . Viable fungal blastoconidia induced different CL-responses: (1) virulent C . albicans and C . tropicalis induced a low CL-response, (2) less virulent C . glabrata, C . parapsilosis and C . krusei induced CL-responses which reflected strain-specific differences, and (3) viable blastoconidia of attenuated C . guilliermondii uniformly caused a high PMN CL-response . After the CL-measurement, the killing of Candida by PMNs was assayed . High CL-response could be correlated to an augmented killing of the yeasts . These results were discussed with respect to a factor of fungal virulence, the secretion of acid proteinase.

Microbiol Rev, 1992 Mar, 56(1), 229 - 64
Recent evidence for evolution of the genetic code; Osawa S et al.; The genetic code, formerly thought to be frozen, is now known to be in a state of evolution . This was first shown in 1979 by Barrell et al . (G . Barrell, A . T . Bankier, and J . Drouin, Nature {London} 282:189-194, 1979), who found that the universal codons AUA (isoleucine) and UGA (stop) coded for methionine and tryptophan, respectively, in human mitochondria . Subsequent studies have shown that UGA codes for tryptophan in Mycoplasma spp . and in all nonplant mitochondria that have been examined . Universal stop codons UAA and UAG code for glutamine in ciliated protozoa (except Euplotes octacarinatus) and in a green alga, Acetabularia . E . octacarinatus uses UAA for stop and UGA for cysteine . Candida species, which are yeasts, use CUG (leucine) for serine . Other departures from the universal code, all in nonplant mitochondria, are CUN (leucine) for threonine (in yeasts), AAA (lysine) for asparagine (in platyhelminths and echinoderms), UAA (stop) for tyrosine (in planaria), and AGR (arginine) for serine (in several animal orders) and for stop (in vertebrates) . We propose that the changes are typically preceded by loss of a codon from all coding sequences in an organism or organelle, often as a result of directional mutation pressure, accompanied by loss of the tRNA that translates the codon . The codon reappears later by conversion of another codon and emergence of a tRNA that translates the reappeared codon with a different assignment . Changes in release factors also contribute to these revised assignments . We also discuss the use of UGA (stop) as a selenocysteine codon and the early history of the code.

Int J Dermatol, 1992 Mar, 31(3), 193 - 5
Eosinophilic pustular folliculitis in patients with acquired immunodeficiency syndrome; Ferrandiz C et al.; Three patients with AIDS had an intensely pruritic eosinophilic pustular folliculitis associated with abundant Pityrosporum yeasts in the follicles, demonstrated by direct microscopy . The excellent response to ketoconazole treatment suggests a possible pathogenic role for Pityrosporum.

Mol Cell Biol, 1992 Mar, 12(3), 1257 - 65
The HeLa Pur factor binds single-stranded DNA at a specific element conserved in gene flanking regions and origins of DNA replication; Bergemann AD et al.; A major site of DNA bending is located 1.6 kb upstream of the P1 transcription start site of the human c-myc gene, near the center of a reported zone of initiation of DNA replication . A repeated, purine-rich element, termed PUR, at the bend site is specifically bound by a protein in HeLa cell nuclear extracts . This protein has specific affinity for the purine-rich single strand of the element . Methylation interference maps a pattern of specific contact points with guanosine bases in a 24-mer oligonucleotide containing the element . UV cross-linking reveals that contact is made by a polypeptide of approximately 28 kDa . The PUR element is present at origins of replication and in gene flanking regions in a variety of eukaryotes from yeasts through humans . The consensus sequence GGNNGAGGGAGARRRR has been derived . This element is present near centers of regions of two mammalian loci (human c-myc and hamster dhfr) recently reported as initiation zones for DNA replication . A 24-mer oligonucleotide representing the hamster dhfr version of the PUR element effectively competes with the human c-myc version for binding to Pur.

Rev Inst Med Trop Sao Paulo, 1992 Mar-Apr, 34(2), 159 - 65
{Preservation of fungi and actinomycetes of medical importance in distilled water}; Rodrigues EG et al.; Several methods have been used for the preservation of fungi, all of them presenting advantages and disadvantages . The choice of the methods depends upon the laboratory availabilities, time of preservation, genetic stability of the cultures and other factors . In this work the results obtained through the utilization of Castellani's method (preservation in distilled water) for the maintenance of 174 strains belonging to the "Micoteca do Instituto de Medicina Tropical de Sao Paulo" are presented . These strains were analyzed after 6, 12, 18 and 24 months, with regard to the percentage of viability taking into consideration the rates of growth and contamination . The smallest percentage of viability occurred in the group of the actinomycetes (50 to 100%) and the largest one in the group of the yeasts (near 100%) . According to other authors, the Castellani's method, besides being simple and economically feasible for small size laboratories, yields good results.

Poult Sci, 1992 Feb, 71(2), 296 - 301
Presence of the diurnal rhythms of monocyte count and macrophage activities in chicks; Kondo Y et al.; The diurnal rhythms of monocyte count in the peripheral blood and the nonspecific activities of peritoneal macrophages were investigated in 6- to 12-wk-old White Leghorn chicks . It was evident that monocyte count and activities of peritoneal macrophages, i.e., phagocytosis and microbicidal activity, showed clear diurnal rhythms . The rhythmic pattern of monocyte count was biphasic, unlike those of humans or mice (which exhibit a monophasic fluctuation pattern) . Diurnal rhythms of phagocytic activity and microbicidal activity also showed two peaks . However, the correspondence of fluctuation patterns between phagocytic activity and microbicidal activity that have been shown in chicken heterophils was not shown in chicken macrophages . This result suggests that the activities of both functions fluctuate independently in chicken macrophages . The patterns of macrophage-nonspecific activities were not in agreement with any rhythmic patterns concerning B cell and T cell activities reported previously . This result may indicate that the accessory functions of macrophage for helper T cell activation are not involved in the formation of the rhythmic patterns of B cell and T cell functions in chicks.

Crit Rev Biotechnol, 1992, 12(1-2), 87 - 132
The production of organic acids; Mattey M; The production of organic acids covers two aspects: first, the metabolic pathways involved in the biosynthesis, and, second, the industrial process strategy adopted . The review seeks to show the underlying biochemical similarities in the biosynthesis of organic acids and the resulting similarities in the commercial processes . Two groups of acids are defined, those with a "long" biosynthetic path from glucose, involving much of the glycolytic pathway and the tricarboxylic acid cycle, and those acids with a "short pathway", essentially a biotransformation of glucose . The regulation of the pathways and the future developments in metabolic control theory and genetic manipulations relating to them are considered . The organisms used industrially are also limited, Aspergillus sp . and Candida yeasts; again the underlying metabolic similarities lead to similar strategies for all the acids discussed.

Eur Neurol, 1992, 32(2), 70 - 3
Chronic Histoplasma capsulatum infection of the central nervous system successfully treated with fluconazole; Tiraboschi I et al.; A 48-year-old man with a long-standing history of communicating hydrocephalus is reported . Ventriculoperitoneal shunting led to clinical improvement, but symptoms recurred despite surgical re-exploration switching the shunt to an atrial drainage . Ten months after the last surgical procedure, an acute myelopathy developed . Concomitant pharyngeal granuloma examination identified Histoplasma capsulatum (Hc) yeasts . Despite initial response to amphotericin B, Hc was isolated from cerebrospinal fluid (CSF), valve reservoir and distal catheter after two courses of therapy . Fluconazole successfully sterilized CSF, but transverse myelopathy persisted unchanged, and shunting was needed to control hydrocephalus.

Annu Rev Physiol, 1992, 54, 579 - 99
Anhydrobiosis; Crowe JH et al.; We believe we have established the major principles governing the stabilization of living cells in the unique condition known as anhydrobiosis . These findings have permitted us to design ways to stabilize membrane vesicles, liposomes, and proteins, and perhaps eventually even intact cells that do not normally survive dehydration . In a complex phenomenon as ancient as anhydrobiosis, one would expect a myriad of adaptations to be required for survival of drying . But the arguments presented here suggest that a single perturbation--synthesis of a disaccharide such as trehalose or sucrose--is sufficient to achieve survival . We hasten to add, however, that it is now certain that additional adaptations are required; for instance, cells containing highly unsaturated lipids may survive drying for a short time, but they are so susceptible to degradation that they survive for a short time only . Thus the interpretation placed on the finding that trehalose can stabilize dry membranes must be regarded from this perspective as well . Nevertheless, we believe that the underlying physical principles governing stability of dry biological materials are universal.

Dermatology, 1992, 184 Suppl 1, 3 - 7
Preclinical data and mode of action of amorolfine; Polak A; Amorolfine is an antifungal showing activity against fungi pathogenic to plants, animals and humans . Amorolfine possesses a broad antifungal spectrum including dermatophytes, yeasts, dimorphic fungi and moulds and is not only fungistatic but fungicidal against most species . Amorolfine interferes with ergosterol biosynthesis at two steps: the delta 14 reduction and the delta 7-8 isomerisation . As a consequence of this inhibition the delta 14 sterol ignosterol is accumulated in the cell membrane and ergosterol is depleted . The cell wall thickness is significantly increased and chitin deposits are included inside and outside . In experimental models of systemic mycosis amorolfine shows no significant activity . This lack of systemic activity may be due to strong protein binding and/or rapid metabolism . In models of superficial fungal infection--trichophytosis and vaginal candidosis--amorolfine has a high activity . On a concentration basis amorolfine is more effective in trichophytosis than naftifine and all azoles tested . Amorolfine clears mycotic foci of trichophytosis in the guinea pig in 10 days, while none of the azoles is able to cure these animals . Tolciclate and terbinafine are the only other substances with a curative effect in these experiments . Amorolfine has a long retention time in the horny layer of the skin . In vaginal candidosis 0.1% amorolfine clears the vagina of viable candida cells in rats.

Annu Rev Biochem, 1992, 61, 897 - 946
Zinc proteins: enzymes, storage proteins, transcription factors, and replication proteins; Coleman JE; In the past five years there has been a great expansion in our knowledge of the role of zinc in the structure and function of proteins . Not only is zinc required for essential catalytic functions in enzymes (more than 300 are known at present), but also it stabilizes and even induces the folding of protein subdomains . The latter functions have been most dramatically illustrated by the discovery of the essential role of zinc in the folding of the DNA-binding domains of eukaryotic transcription factors, including the zinc finger transcription factors, the large family of hormone receptor proteins, and the zinc cluster transcription factors from yeasts . Similar functions are highly probable for the zinc found in the RNA polymerases and the zinc-containing accessory proteins involved in nucleic acid replication . The rapid increase in the number and nature of the proteins in which zinc functions is not unexpected since zinc is the second most abundant trace metal found in eukaryotic organisms, second only to iron . If one subtracts the amount of iron found in hemoglobin, zinc becomes the most abundant trace metal found in the human body.

Arch Vet Pol, 1992, 32(3-4), 5 - 13
The ultrastructure of Pityrosporum pachydermatis; Winiarczyk S; The purpose of the present work was to make a thorough study of the ultrastructure of yeasts Pityrosporum pachydermatis under transmission electron microscope . The study revealed an additional layer, that has not been described so far, situated outside the cell wall . Because its presence could be proved exclusively in the specimens prepared by means of a method which permits minimal washing out of lipids from biological material (dehydration by means of hexylene glycol and immersion in the resin of low viscosity), one can assume, that lipids are one of the components of this outer layer and they bind its structure together.

Australas J Dermatol, 1992, 33(3), 159 - 63
Dermatophyte and non-dermatophyte onychomycosis in Singapore; Lim JT et al.; Onychomycosis is caused by dermatophytes, moulds and yeasts . It is important to identify the non-dermatophyte moulds as they are resistant to the usual anti-fungals . A prospective study was undertaken in the National Skin Centre, Singapore to study the pattern of dermatophyte and non-dermatophyte onychomycosis . 53 male and 47 female patients seen between June 1990 and June 1991 were entered into the study . Direct microscopy was done and the nail clippings were cultured . Toe and finger nails were equally infected . Dermatophytes were isolated from 21 patients namely, T . rubrum (12/21), T . interdigitale (5/21), T . mentagrophytes (3/21) and T . violaceum (1/21) . Candida onychomycosis occurred in 39 patients and was caused by C . albicans (38/39) and C . parapsilosis (1/39) . 37/39 patients had associated paronychia . 5 types of moulds were isolated from 12 patients, namely Fusarium species (6/12), Aspergillus species (3/12), S . brevicaulis (1/12), Aureobasilium species (1/12) and Penicillium species (1/12) . Although the clinical pattern and microscopy may predict the type of organisms, in practice this is difficult . Only cultures were confirmatory . 28% (28/100) had negative cultures despite a positive microscopy, and moulds (12/100) grown might be contaminants rather than pathogens.

Biosystems, 1992, 28(1-3), 117 - 25
Detecting morphological convergence in true fungi, using 18S rRNA gene sequence data; Berbee ML et al.; For the true fungi, phylogenetic relationships inferred from 18S ribosomal DNA sequence data agree with morphology when (1) the fungi exhibit diagnostic morphological characters, (2) the sequence-based phylogenetic groups are statistically supported, and (3) the ribosomal DNA evolves at roughly the same rate in the lineages being compared . 18S ribosomal RNA gene sequence data and biochemical data provide a congruent definition of true fungi . Sequence data support the traditional fungal subdivisions Ascomycotina and Basidiomycotina . In conflict with morphology, some zygomycetes group with chytrid water molds rather than with other terrestrial fungi, possibly owing to unequal rates of nucleotide substitutions among zygomycete lineages . Within the ascomycetes, the taxonomic consequence of simple or reduced morphology has been a proliferation of mutually incongruent classification systems . Sequence data provide plausible resolution of relationships for some cases where reduced morphology has created confusion . For example, phylogenetic trees from rDNA indicate that those morphologically simple ascomycetes classified as yeasts are polyphyletic and that forcible spore discharge was lost convergently from three lineages of ascomycetes producing flask-like fruiting bodies.

Mycopathologia, 1991 Dec, 116(3), 155 - 8
Prevalence of superficial mycoses in the Aosta Valley region of Italy from 1984 to 1989; Dal Tio R et al.; This report represents the results of the culture tests for fungi carried out in the Aosta Valley region of Italy from 1984 to 1989 . The pathological material consisted of cutaneous scales, hair and nails . It was possible to identify 91.8% of the isolates: out of these, 36.1% were dermatophytes, 21% were Aspergillus spp, 15.6% Deuteromycetes, 15% yeasts and 4.1% members of the family Mucoraceae . The resulting data indicate that Microsporum canis was the most wide-spread species (73.7%) among the isolated dermatophytes and keratinophilic fungi (Trichophyton rubrum, T . mentagrophytes, Epidermophyton floccosum and Scopulariopsis brevicaulis): 87.5% were isolated from hair and 65.9% from the glabrous skin . The high prevalence of M . canis as the cause of dermatomycoses is discussed . Lastly the use made of the Regional Hospital's Mycology Department by health workers is discussed.

Nucleic Acids Res, 1991 Nov 25, 19(22), 6221 - 6
RNase P RNA in Candida glabrata mitochondria is transcribed with substrate tRNAs; Shu HH et al.; The biosynthesis of some mitochondrial enzymes requires contributions of both the mitochondrial and nuclear genomes . The ribonucleoprotein enzyme Ribonuclease P (RNase P) is composed of a mitochondrial encoded RNA and nuclear coded protein in many yeasts, including C . glabrata . We have determined that there are at least two sites of transcription initiation that contribute to the expression of the mitochondrial RNase P RNA . A nonanucleotide promoter sequence is located upstream of the initiator tRNA while the other site of initiation of transcription is at an undetermined upstream site . An analysis of the transcripts from the region of the RNase P gene demonstrates directly that the RNase P RNA is present in large primary transcripts and located between the precursors to the initiator tRNAf(Met) and tRNA(Pro) genes . Thus this enzyme subunit is synthesized with some of its substrate tRNAs . An activity with cleavage site specificity like a previously described endonuclease that cleaves near the 3' end of tRNAs, RNase P activity and one or more additional endonucleases or exonucleases not described previously are required to convert the primary transcript to its final functional RNAs.

J Mol Biol, 1991 Oct 20, 221(4), 1191 - 207
Model for the interaction of DNA junctions and resolving enzymes; Bhattacharyya A et al.; Four-way DNA junctions are thought to be important intermediates in a number of recombination processes . Resolution of these junctions occurs by cleavage of two strands of DNA to generate two duplex molecules . The interaction between DNA junctions and resolving enzymes appears to be largely structure-specific, reflecting a molecular recognition on a significant scale . We propose a working model for this interaction that takes account of the present state of knowledge of the structure of the DNA junction, and the substrate requirements of the enzymes . We note that three different enzymes introduce cleavages at phosphodiester bonds that are presented on one side of the molecule, suggesting that the enzymes selectively interact with this face of the junction . By forcing a junction of constant sequence to adopt one or other of the two possible antiparallel isomers, we show that the junction is cleaved in such a way as to suggest a constant mode of interaction with the protein that is dependent on structure rather than sequence . We propose that the feature that is recognized is a mutual inclination of two DNA helices at approximately 120 degrees . We show that a number of DNA substrates that contain similar inclined helices, such as a three-way junction, bulged duplexes and a duplex that is curved because of repeated runs of oligoadenine sequences, are each cleaved by phage T4 endonuclease VII . This mode of DNA-protein interaction could be significant in either recombination or DNA repair processes.

Dtsch Tierarztl Wochenschr, 1991 Oct, 98(10), 381 - 3
{The occurrence and metabolism of physiologically normal-appearing hydrocarbons in agricultural animals}; Lusky K et al.; Carbohydrates are determined in food from animals as result of contamination . Therefore they are a risk for the consumer and may impair the quality of the products . There are differences in the pattern of residues between the groups of carbohydrates . Especially the saturated carbohydrates indicate heavy differences . Aromatic carbohydrates have an equalized level . These relationships are examined by tests with n-alkane-yeasts-feeding of pigs.

Biochim Biophys Acta, 1991 Aug 6, 1074(3), 392 - 7
Penicillium chrysogenum extracellular acid phosphatase: purification and biochemical characterization; Haas H et al.; An extracellular acid phosphatase (EC 3.1.3.2) from crude culture filtrate of Penicillium chrysogenum was purified to homogeneity using high-performance ion-exchange chromatography and size-exclusion chromatography . SDS-PAGE of the purified enzyme exhibited a single stained band at an Mr of approx . 57,000 . The mobility of the native enzyme indicated the Mr to be 50,000, implying that the active form is a monomer . The isoelectric point of the enzyme was estimated to be 6.2 by isoelectric focusing . Like acid phosphatases from several yeasts and fungi the Penicillium enzyme was a glycoprotein . Removal of carbohydrate resulted in a protein band with an Mr of 50,000 as estimated by SDS-PAGE, suggesting that 12% of the mass of the enzyme was carbohydrate . The enzyme was catalytically active at temperatures ranging from 20 degrees C to 65 degrees C with a maximum activity at 60 degrees C and the pH optimum was at 5.5 . The Michaelis constant of the enzyme for p-nitrophenyl phosphate was 0.11 mM and it was inhibited competitively by inorganic phosphate (ki = 0.42 mM).

Mol Cell Biol, 1991 Aug, 11(8), 3823 - 34
Chromosome structure: DNA nucleotide sequence elements of a subset of the minichromosomes of the protozoan Trypanosoma brucei; Weiden M et al.; The genome of the protozoan Trypanosoma brucei contains a set of about 100 minichromosomes of about 50 to 150 kb in size . The small size of these chromosomes, their involvement in antigenic variation, and their mitotic stability make them ideal candidates for a structural analysis of protozoan chromosomes and their telomeres . We show that a subset of the minichromosomes is composed predominantly of simple-sequence DNA, with over 90% of the length of the minichromosome consisting of a tandem array of 177-bp repeats, indicating that these molecules have limited protein-coding capacity . Proceeding from the tip of the telomere to a chromosome internal position, a subset of the minichromosomes contained the GGGTTA telomere repeat, a 29-bp telomere-derived repeat, a region containing 74-bp G + C-rich direct repeats separated by approximately 155 bp of A + T-rich DNA that has a bent character, and 50 to 150 kb of the 177-bp repeat . Several of the minichromosome-derived telomeres did not encode protein-coding genes, indicating that the repertoire of telomeric variant cell surface glycoprotein genes is restricted to some telomeres only . The telomere organization in trypanosomes shares striking similarities to the organization of telomeres and subtelomeres in humans, yeasts, and plasmodia . An electron microscopic analysis of the minichromosomes showed that they are linear molecules without abnormal structures in the main body of the chromosome . The structure of replicating molecules indicated that minichromosomes probably have a single bidirectional origin of replication located in the body of the chromosome . We propose a model for the structure of the trypanosome minichromosomes.

Rev Infect Dis, 1991 Jul-Aug, 13(4), 592 - 9
Response to empiric amphotericin B during antileukemic therapy-induced granulocytopenia; Karp JE et al.; We analyzed the initial and overall responses to empiric therapy with amphotericin B as they related to the rate of occurrence and the type of fungal infection and colonization during 264 consecutive episodes of prolonged, profound, chemotherapy-induced bone marrow aplasia (greater than 30 days, less than 100 polymorphonuclear leukocytes/mm3) in 160 adults with acute leukemia . Amphotericin B was administered during 248 (94%) of these granulocytopenic episodes; in 68 cases the drug was given because of documented infection with yeasts or filamentous fungi (DFI), and in 180 cases it was given because of refractory fever without DFI . The frequency of an initial response in patients with DFI (60%) was similar to that in non-DFI-infected patients (61%) . Both the initial response rate and the overall survival rate were significantly decreased when therapy with amphotericin B was not initiated empirically before documentation of filamentous DFI . Given the comparatively high rates of initial and overall response (the latter being 74% and 71% for DFI and non-DFI, respectively) and the lack of alternative fungicidal agents, our data support prompt empiric treatment with amphotericin B for refractory fever in adults with acute leukemia who are compromised by severe, therapy-induced granulocytopenia.

Mycoses, 1991 Jul-Aug, 34(7-8), 339 - 44
Fungal infection as a cause of skin disease in the eastern province of Saudi Arabia: tinea pedis and tinea manuum; al-Sogair SM et al.; In the period between April 1984 and April 1988 a total of 4294 clinically suspected cases of dermatomycoses were examined for causative fungi . Of these cases 680 were suspected as tinea pedis and tinea manuum . These cases belonged to 21 different nationalities . Both sexes were represented and 649 cases (95.4%) were adults while 31 (4.6%) were prepubertals . Clinical diagnosis was confirmed by direct microscopy in 505 cases or 74.4% (485 adults and 20 prepubertal children) . Out of these, 504 cases were positive on culture and yielded 516 isolates . Candida species and other yeasts were responsible for 88.9% and dermatophytes for 11.1% of these infections . The interdigital type of lesions was the most common type (91.9% of all infections) followed by the hyperkeratotic scaling type (6.9%) while the acute inflammatory type was only 1.2% of infections . Treatment showed satisfactory results in most of the cases.

Mycoses, 1991 Jul-Aug, 34(7-8), 333 - 7
Fungal infection as a cause of skin disease in the eastern province of Saudi Arabia: prevailing fungi and pattern of infection; al-Sogair SM et al.; A total of 4,294 clinically suspected cases of dermatomycoses belonging to 26 different nationalities were examined between April 1984 and April 1988 . Fungi were demonstrated in routine potassium hydroxide/dimethyl sulfoxide mount in 3,814 cases (88.8%) and the etiology was determined by culture in 2,458 cases (57.2%) . Tinea versicolor was the predominant fungal infection (30.9% of all infections) . Onychomycosis and paronychia ranked second in prevalence (16.8%) . Candidal onychomycosis was the most common type of infection . Scalp ringworm among children ranked third (15.3%), Microsporum canis was the main etiologic agent . Tinea pedis and tinea manuum ranked fourth in prevalence (13.2%) . Tinea corporis represented 10.7% of infections and M . canis was the main agent . Tinea cruris accounted for 8.7% of infections and Epidermophyton floccosum was the most common agent . Cutaneous candidosis constituted 4.3% of infections . White piedra was seen in 6 cases (0.16%) . Yeasts were proved not to be unimportant as a cause of disease of skin and nail in our study.

FASEB J, 1991 Jun, 5(9), 2237 - 42
Molecular studies of the uncoupling protein; Ricquier D et al.; The uncoupling protein (UCP) is a proton/anion transporter found in the inner mitochondrial membrane of brown adipocyte . Although UCP has not been detected in mitochondria from any other tissue, it shares structural and catalytic properties with several other mitochondrial carrier proteins . Although UCP was discovered only recently it is one of the most extensively studied mitochondrial carrier proteins . Many tools useful in research on UCP have been developed such as antibodies and cDNAs corresponding to UCP of several animal species . More recently, the mouse, rat, and human genes encoding for UCP have been isolated and sequenced . The availability of these various tools has led to several significant observations . UCP gene expression is strongly controlled at the level of transcription by signals that are activated after the stimulation of brown adipocytes by norepinephrine . The comparison of UCP gene with the genes encoding the adenine nucleotide translocator revealed the existence of structural and evolutionary homologies . Moreover, in humans the UCP gene and one form of adenine nucleotide translocator gene are located on the same chromosome . Recently, the expression of functional UCP in various heterologous systems was achieved (Xenopus oocytes, CHO cells, yeasts) . These data will facilitate studies of the structure/function relationship in UCP (identification of residues involved in H+ transport, Cl- transport, nucleotide binding, mitochondrial targeting...) . Another aspect of the present research on UCP is the understanding of mechanisms that control the UCP gene and the differentiated commitment of adipose precursor cells to thermogenic brown adipocytes . The multifaceted aspects of research on UCP make this protein interesting in areas of research as different as studies of ion translocating mechanisms, cellular specificity of gene transcription, control of gene expression by neuromediators, adipocyte differentiation, and the pharmacological treatment of obesity.

Gig Sanit, 1991 Jun, (6), 12 - 4
{Dynamics of the changes in the basic protective systems of an organism under the influence of unicellular protein}; Iakimova EP et al.; In the present work the results of study in functional and metabolic reactivity of protective systems are summarized . The early stages of the biological actions of protein-vitamin concentrates obtained from ethanol utilizing yeasts by means of unified test-system were studied.

Biochim Biophys Acta, 1991 May 17, 1092(3), 350 - 7
Recent progress in characterization of protein kinase cascades for phosphorylation of ribosomal protein S6; Sturgill TW et al.; Ribosomal protein S6 is phosphorylated in response to mitogens by activation of one or more protein kinase cascades . Phosphorylation of S6 in vivo is catalyzed by (at least) two distinct mitogen-activated S6 kinase families distinguishable by size, the 70 kDa and 90 kDa S6 kinases . Both S6 kinases are activated by serine/threonine phosphorylation . Members of each family have been cloned . The 90 kDa S6 kinases are activated more rapidly than the 70 kDa S6 kinase, and may have other intracellular targets . The 70 kDa S6 kinase is relatively specific for 40 S ribosomal subunits . No kinase capable of activating the 70 kDa S6 kinase has been identified . Members of the 90 kDa S6 kinases are activated in vitro by 42 kDa and 44 kDa MAP kinases, which are in turn activated by mitogen-dependent activators . The pathways for mitogen-stimulated S6 phosphorylation are discussed.

Infect Immun, 1991 May, 59(5), 1639 - 46
Infection of P388D1 macrophages and respiratory epithelial cells by Histoplasma capsulatum: selection of avirulent variants and their potential role in persistent histoplasmosis; Eissenberg LG et al.; We evaluated P388D1 macrophagelike cells as model host cells for studying the intracellular survival and strain-specific virulence of Histoplasma capsulatum . Previously characterized strains which were virulent for mice destroyed monolayers of these cells within a few days . In contrast, related avirulent "smooth" variants failed to do so even after 20 days, although they persisted within P388D1 cells for at least 7 days . On the basis of this observation, we developed a quantitative radiolabel assay to use as an initial screen for virulence . Another cell type lining the respiratory tract was then examined as a potential host for H . capsulatum . Hamster trachea epithelial (HTE) cells readily internalized a variety of strains lacking alpha-(1,3)-glucan in their cell walls; however, the tracheal cells were only rarely infected by organisms possessing this polysaccharide . We subsequently inoculated HTE cells with alpha-(1,3)-glucan-positive strains and enriched for the few yeasts infecting these cells . The progeny resembled smooth variants in terms of colony morphology, the absence of alpha-(1,3)-glucan in their cell walls, and their inability to kill macrophages . Did the HTE cells select for these variant yeasts from the parent inoculum or instigate a change from the parental phenotype? Following a 3-h uptake period, only 2% of the ingested yeasts lacked alpha-(1,3)-glucan . One day later, nearly half of the intracellular organisms lacked this polymer . This rapid conversion of a large proportion of the inoculum suggests some type of environmentally triggered change, perhaps analogous to phase variation seen in many other pathogens . Infection of epithelial cells or some other nonprofessional phagocyte during natural histoplasmosis might give rise to similar variants, thus establishing a reservoir of organisms capable of causing chronic or latent infections.

Boll Chim Farm, 1991 May, 130(5), 166 - 8
In vitro activity of phenyl-O,N,N-azoxycyanide towards Candida spp . and Torulopsis spp; Tullio V et al.; Antifungal activity in vitro of phenyl-0,N,N-azoxycyanide has been tested against some strains of Candida spp . and Torulopsis spp . freshly isolated from humans . The results indicate good activity versus all yeasts tested . These data suggest that the azoxycyanide function could be used to obtain derivatives with potent anticandidal action, if conveniently vehicled by lypophylic compounds.

Acta Gastroenterol Belg, 1991 May-Aug, 54(3-4), 242 - 7
{The virus of non-A, non-B hepatitis . Serological diagnosis}; Donea-Debroise B; Persistence of viral post-transfusion hepatitis together with epidemiological data led to identify 3 forms of clinical non A non B hepatitis: enteric, post-transfusional and sporadic hepatitis . Two groups of viruses are responsible for this pathology; they are designated as HEV (Hepatitis E Virus) and HCV (Hepatitis C Virus) . HEV described by D . Bradley is a 27 to 34 nm . non enveloped particle containing a single strand RNA and belonging to the calicivividae family . HCV described by M . Houghton is a 50 to 60 nm . enveloped virus containing a 10,000 nucleotide long single strand RNA who belongs to the Flaviviridae family . The serological diagnosis is based on the detection of antibodies against the C100-3 antigen, a 363 amino acid recombinant protein produced in yeasts . Reactive samples are to be confirmed for antibodies specificity by using a neutralization assay in the presence of the same antigenic material . Alternatively, it is also possible to identify some sequences of viral genome in the serum, after amplification by the technic of Polymerase Chain Reaction (PCR).

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