Nat Rev Mol Cell Biol, 2002 Dec, 3(12), 919 - 31 Sorting out the cellular functions of sorting nexins; Worby CA et al.; Sorting nexins are a diverse group of cellular trafficking proteins that are unified by the presence of a phospholipid-binding motif - the PX domain . The ability of these proteins to bind specific phospholipids, as well as their propensity to form protein-protein complexes, points to a role for these proteins in membrane trafficking and protein sorting . It will be interesting to determine whether the various sorting nexins have specialized or generalized roles in protein trafficking. Food Addit Contam, 2002 Nov, 19(11), 1081 - 90 Post-harvest storage of corn: effect of beginning moisture content on mycoflora and fumonisin contamination; Ono EY et al.; The effect of storage on mycoflora profile was monitored bimonthly in 36 corn (Zea mays L.) samples, dividing the same sample into groups dried to 11 and 14% moisture content (1008 analysis) . These groups were further subdivided based on the initial total count (moulds and yeasts) up to 10(4) CFU g(-1) (12 samples, range 1.6 x 10(4) to 9.0 x 10(4), mean 3.8 x 10(4) CFU g(-1)) and up to 10(5) CFU g(-1) (24 samples, range 1.0 x 10(5) to 5.0 x 10(5), mean 2.7 x 10(5) CFU g(-1)) . In the corn group dried to 11%, the fumonisin content was analysed at the initial stage (freshly harvested) and at the end of 12-month storage . Fusarium spp . and Penicillium spp . prevailed at the freshly harvested stage (100%), maintaining this profile throughout 12 months, in corn dried to both 11 and 14% . Cladosporium spp., Aspergillus spp . and Phoma spp . were also detected at lower frequencies during the storage . Fusarium spp . and the total fungal colony count during 12-month storage carried out with samples dried to 11 or 14% moisture content were statistically evaluated using ANOVA for randomized complete block design . The correlation between storage time and Fusarium spp . and total fungal colony count data was analysed by Pearson's correlation test . There was no difference in Fusarium spp . and total counts in the 10(4) CFU g(-1) initial total count group throughout the storage time (p < 0.05) . There was a negative correlation between fungal population and storage time (p < 0.05) in the 10(5) CFU g(-1) initial total count group . Fumonisins were detected in all freshly harvested corn, at a mean concentration of 9.9 +/- 6.0 micro g g(-1) (range 0.74-22.6 micro g g(-)1) . These values did not change in the 12-month stored corn (mean of 9.9 +/- 5.8 micro g g(-1), range 0.81-23.7 micro g g(-1)) . These post harvest data indicated the importance of moisture content at the crop harvesting/predrying stage to control fungal growth and further fumonisin production. Eukaryot Cell, 2002 Jun, 1(3), 378 - 90 A G-protein beta subunit required for sexual and vegetative development and maintenance of normal G alpha protein levels in Neurospora crassa; Yang Q et al.; The genome of the filamentous fungus Neurospora crassa contains a single gene encoding a heterotrimeric G-protein beta subunit, gnb-1 . The predicted GNB-1 protein sequence is most identical to G beta proteins from the filamentous fungi Cryphonectria parasitica and Aspergillus nidulans . N . crassa GNB-1 is also 65% identical to the human GNB-1 protein but only 38 and 45% identical to G beta proteins from budding and fission yeasts . Previous studies in animal and fungal systems have elucidated phenotypes of G beta null mutants, but little is known about the effects of G beta loss on G alpha levels . In this study, we analyzed a gnb-1 deletion mutant for cellular phenotypes and levels of the three G alpha proteins . Delta gnb-1 strains are female-sterile, with production of aberrant fertilized reproductive structures . Delta gnb-1 strains conidiate more profusely and have altered mass on solid medium . Loss of gnb-1 leads to inappropriate conidiation and expression of a conidiation-specific gene during growth in submerged culture . Intracellular cyclic AMP levels are reduced by 60% in vegetative plate cultures of delta gnb-1 mutants . Loss of gnb-1 leads to lower levels of the three G alpha proteins under a variety of conditions . Analysis of transcript levels for the gna-1 and gna-2 G alpha genes in submerged cultures indicates that regulation of G alpha protein levels by gnb-1 is posttranscriptional . The results suggest that GNB-1 directly regulates apical extension rate and mass accumulation . In contrast, many other delta gnb-1 phenotypes, including female sterility and defective conidiation, can be explained by altered levels of the three N . crassa G alpha proteins. Mikrobiologiia, 2002 Sep-Oct, 71(5), 667 - 74 {The structure of the micromycete complexes of oligotrophic peat deposits in the southern taiga subzone of West Siberia}; Golovchenko AV et al.; The analysis of the micromycete complexes of oligotrophic peat deposits in the Vasyugan Marsh by direct count and culture methods showed that the micromycete carbon comprises no more than 3% of the total peat carbon and that the microscopic fungal biomass varies from 2 to 13 tons/hectare, depending on the season and the peat deposit thickness . Fungal spores were found in all layers of the peat deposits, whereas the mycelium was found only in the active peat layer . The high abundance of eukaryotic cells in the peats was due to the presence of yeastlike cells rather than fungal spores . Analyses by culture methods showed that micromycetes were present in all peat layers and that their abundance tended to decrease with depth, except for yeasts, which were uniformly distributed in a vertical direction . The micromycete complexes of the peat deposits were similar in the diversity and abundance of dominant species but differed in the composition of minor species . Peat yeasts were dominated by ascomycetes. Trends Genet, 2002 Dec, 18(12), 636 - 42 Intracellular mRNA localization: motors move messages; Tekotte H et al.; Intracellular mRNA localization is a common mechanism of post-transcriptional regulation of gene expression . In a wide range of organisms, mRNA localization coupled with translational regulation target the proteins to their site of function . Here, we describe recent exciting evidence that some mRNAs are transported as particles along the cytoskeleton by the molecular motors dynein, kinesin or myosin . We discuss the key questions of how localized mRNAs might be linked to motors and what determines their cytoplasmic destinations. Trends Cell Biol, 2002 Nov, 12(11), 509 - 16 Checking on the fork: the DNA-replication stress-response pathway; Osborn AJ et al.; To ensure the fidelity of DNA replication, cells activate a stress-response pathway when DNA replication is perturbed . This pathway regulates not only progress through the cell cycle but also transcription, apoptosis, DNA repair/recombination and DNA replication itself . Mounting evidence has suggested that this pathway is important for the maintenance of genomic integrity . Here, we discuss recent findings about how this pathway is activated by replication stress and how it regulates the DNA-replication machinery to alleviate the stress. Curr Biol, 2002 Nov 19, 12(22), 1908 - 18 Caenorhabditis elegans HUS-1 is a DNA damage checkpoint protein required for genome stability and EGL-1-mediated apoptosis; Hofmann ER et al.; BACKGROUND: The inability to efficiently repair DNA damage or remove cells with severely damaged genomes has been linked to several human cancers . Studies in yeasts and mammals have identified several genes that are required for proper activation of cell cycle checkpoints following various types of DNA damage . However, in metazoans, DNA damage can induce apoptosis as well . How DNA damage activates the apoptotic machinery is not fully understood . RESULTS: We demonstrate here that the Caenorhabditis elegans gene hus-1 is required for DNA damage-induced cell cycle arrest and apoptosis . Following DNA damage, HUS-1 relocalizes and forms distinct foci that overlap with chromatin . Relocalization does not require the novel checkpoint protein RAD-5; rather, relocalization appears more frequently in rad-5 mutants, suggesting that RAD-5 plays a role in repair . HUS-1 is required for genome stability, as demonstrated by increased frequency of spontaneous mutations, chromosome nondisjunction, and telomere shortening . Finally, we show that DNA damage increases expression of the proapoptotic gene egl-1, a response that requires hus-1 and the p53 homolog cep-1 . CONCLUSIONS: Our findings suggest that the RAD-5 checkpoint protein is not required for HUS-1 to relocalize following DNA damage . Furthermore, our studies reveal a new function of HUS-1 in the prevention of telomere shortening and mortalization of germ cells . DNA damage-induced germ cell death is abrogated in hus-1 mutants, in part, due to the inability of these mutants to activate egl-1 transcription in a cep-1/p53-dependent manner . Thus, HUS-1 is required for p53-dependent activation of a BH3 domain protein in C . elegans. J Nat Prod, 2002 Nov, 65(11), 1712 - 4 Aldehyde dehydrogenase inhibitors from the mushroom Clitocybe clavipes; Kawagishi H et al.; Five fatty acid derivatives including three novel compounds were isolated from the mushroom Clitocybe clavipe . Their structures were elucidated by spectral analyses . These compounds inhibited aldehyde dehydrogenase in vitro. Nucleosides Nucleotides Nucleic Acids, 2002 Jun-Jul, 21(6-7), 449 - 62 Analysis of relative positions of ribonucleotide bases in a crystal structure of ribosome; Takasu A et al.; Relative positions of bases to bases in a crystal structure of ribosome were analyzed extensively . It was found that there is no clear relation between bases apart more than 15 A and, thus, the relative location of bases can be analyzed within 15 A of the reference bases . As for base pairing, major positioning was found to be due to the Watson-Crick type base pairs . Some other positions corresponding to non-Watson-Crick type base pairs were also found in some extents . As for base-base stacking, it was observed that the bases stacked to adenine base are dispersive . It was found that less non-Watson-Crick base pairs was found close to the protein binding site, suggesting that the protein components have a tendency to bind to the regular stem structures . The database of relative location of bases must be useful for improvement of structural determination and structural modeling systems. Biochem Soc Trans, 2002 Nov, 30(Pt 6), 1047 - 50 Regulation of lipid accumulation in oleaginous micro-organisms; Ratledge C; A small number of eukaryotic micro-organisms, the oleaginous species, can accumulate triacylglycerols as cellular storage lipids, sometimes up to 70% of the biomass . Some of these lipids, particularly those containing high proportions of polyunsaturated fatty acids of nutritional and dietary importance, are now in commercial production; these are known as single-cell oils . The biochemistry of lipid accumulation has been investigated in yeasts and filamentous fungi and can now be described in some detail: lipid accumulation is triggered by cells exhausting nitrogen from the culture medium, but glucose continues to be assimilated . Activity of isocitrate dehydrogenase within the mitochondrion, however, now slows or even stops due to the diminution of AMP within the cells . This leads to the accumulation of citrate, which is transported into the cytosol and cleaved to acetyl-CoA by ATP:citrate lyase, an enzyme that does not occur in non-oleaginous species . This enzyme is therefore essential for lipid accumulation . The presence of this enzyme does not, however, explain why different species of oleaginous micro-organisms have different capacities for lipid accumulation . The extent of lipid accumulation is considered to be controlled by the activity of malic enzyme (ME), which acts as the sole source of NADPH for fatty acid synthase (FAS) . If ME is inhibited, or genetically disabled, then lipid accumulation is very low . There is no general pool of NADPH which can otherwise be used by FAS . The stability of ME is therefore crucial and it is proposed that ME is physically attached to FAS as part of the lipogenic metabolon . ME activity correlates closely with lipid accumulation in two filamentous fungi, Mucor circinelloides and Mortierella alpina . When ME ceases to be active, lipid accumulation also stops . No other enzyme activity shows such a correlation. Curr Opin Investig Drugs, 2002 Oct, 3(10), 1437 - 45 DNA binding compounds targeting fungal pathogens: an emerging concept in the discovery of novel antifungal agents; Lou L et al.; With the urgent need for novel agents to combat emerging fungal resistance to existing drugs, activity in the exploration of small molecule DNA binders has increased . Recently, selected cationic heterocyclic compounds belonging to a broad class of molecules known to bind to the minor groove of DNA were revealed to have potent antifungal activity . These molecules are different from the conventional DNA-interacting drugs such as topoisomerase inhibitors, DNA alkylators or intercalators . Selected compounds are fungicidal towards a variety of pathogenic yeasts and molds, and a selected lead compound is efficacious in a mouse model of systemic candidosis. Mutat Res, 2002 Nov 30, 509(1-2), 35 - 47 RecQ helicases and cellular responses to DNA damage; Wu L et al.; The faithful replication of the genome is essential for the survival of all organisms . It is not surprising therefore that numerous mechanisms have evolved to ensure that duplication of the genome occurs with only minimal risk of mutation induction . One mechanism of genome destabilization is replication fork demise, which can occur when a translocating fork meets a lesion or adduct in the template . Indeed, the collapse of replication forks has been suggested to occur in every replicative cell cycle making this a potentially significant problem for all proliferating cells . The RecQ helicases, which are essential for the maintenance of genome stability, are thought to function during DNA replication . In particular, RecQ helicase mutants display replication defects and have phenotypes consistent with an inability to efficiently reinitiate replication following replication fork demise . Here, we review some current models for how replication fork repair might be effected, and discuss potential roles for RecQ helicases in this process. EMBO J, 2002 Nov 15, 21(22), 6162 - 73 Nutrient control of gene expression in Drosophila: microarray analysis of starvation and sugar-dependent response; Zinke I et al.; We have identified genes regulated by starvation and sugar signals in Drosophila larvae using whole-genome microarrays . Based on expression profiles in the two nutrient conditions, they were organized into different categories that reflect distinct physiological pathways mediating sugar and fat metabolism, and cell growth . In the category of genes regulated in sugar-fed, but not in starved, animals, there is an upregulation of genes encoding key enzymes of the fat biosynthesis pathway and a downregulation of genes encoding lipases . The highest and earliest activated gene upon sugar ingestion is sugarbabe, a zinc finger protein that is induced in the gut and the fat body . Identification of potential targets using microarrays suggests that sugarbabe functions to repress genes involved in dietary fat breakdown and absorption . The current analysis provides a basis for studying the genetic mechanisms underlying nutrient signalling. Microsc Res Tech, 2002 Nov 15, 59(4), 325 - 30 Lifespan extension by caloric restriction: an aspect of energy metabolism; Yamaza H et al.; Caloric restriction (CR) may retard aging processes and extend lifespan in organisms by altering energy-metabolic pathways . In CR rodents, glucose influx into tissues is not reduced, as compared with control animals fed ad libitum (AL), although plasma concentrations of glucose and insulin are lower . Gene expression profiles in rodents have suggested that CR promotes gluconeogenesis and fatty acid biosynthesis in skeletal muscle . In the liver, CR promotes gluconeogenesis but decreases fatty acid synthesis and glycolysis . In lower organisms such as yeasts and nematodes, incomplete blocks in steps of insulin/insulin-like growth factor-1 (IGF-1) signal pathway extend lifespan . The life-prolonging effect of CR in yeasts requires NPT1 and SIR2 genes, both of which relate to sensing energy status and silencing genes . These findings stress the substantial role of energy metabolism on CR . Future studies on metabolic adaptation and gene silencing with regard to lower caloric intake will be warranted to understand the mechanisms of the anti-aging and life-prolonging effects of CR . Exp Cell Res, 2002 Nov 1, 280(2), 212 - 21 Ubc9 is essential for viability of higher eukaryotic cells; Hayashi T et al.; Ubc9 is an enzyme involved in the conjugation of SUMO-1 (small ubiquitin related modifier 1) to target proteins . The SUMO-1 conjugation system is well conserved from yeasts to higher eukaryotes, but many SUMO-1 target proteins reported recently in higher eukaryotic cells, including IkappaBalpha, MDM2, p53, and PML, are not present in yeasts . To determine the physiological roles of SUMO-1 conjugation in higher eukaryotic cells, we constructed a conditional UBC9 mutant of chicken DT40 cells containing the UBC9 transgene under control of a tetracycline-repressible promoter and characterized their loss of function phenotypes . Ubc9 disappeared 3 days after the addition of tetracycline and the increase in viable cell number stopped 4 days after the addition of drug . In contrast to the cases of ubc9 mutants of budding and fission yeasts, which show defects in progression of G2 or early M phase and in chromosome segregation, respectively, we did not observe accumulation of cells in G2/M phase or a considerable increase in the frequency of chromosome missegregation upon depletion of Ubc9 but we did observe an increase in the number of cells containing multiple nuclei, indicating defects in cytokinesis . A considerable portion of the Ubc9-depleted cell population was committed to apoptosis without accumulating in a specific phase of the cell cycle, suggesting that chromosome damages are accumulated in Ubc9-depleted cells, and apoptosis is triggered without activating checkpoint mechanisms under conditions of SUMO-1 conjugation system impairment. Genetics, 2002 Oct, 162(2), 977 - 85 Incorporation of large heterologies into heteroduplex DNA during double-strand-break repair in mouse cells; Raynard SJ et al.; In this study, the formation and repair of large (>1 kb) insertion/deletion (I/D) heterologies during double-strand-break repair (DSBR) was investigated using a gene-targeting assay that permits efficient recovery of sequence insertion events at the haploid chromosomal immunoglobulin (Ig) mu-locus in mouse hybridoma cells . The results revealed that (i) large I/D heterologies were generated on one or both sides of the DSB and, in some cases, formed symmetrically in both homology regions; (ii) large I/D heterologies did not negatively affect the gene targeting frequency; and (iii) prior to DNA replication, the large I/D heterologies were rectified. J Cell Biochem, 2002, 87(3), 258 - 65 Roles for cytoplasmic polyadenylation in cell cycle regulation; Read RL et al.; Polyadenylation of eukaryotic mRNAs in the nucleus promotes their translation following export to the cytoplasm and is an important determinant of mRNA stability . An additional level of control of gene expression is provided by cytoplasmic polyadenylation, which activates translation of a number of mRNAs important in orchestrating cell cycle events in oocytes . Recent studies indicate that cytoplasmic polyadenylation may be a mechanism of translational activation that is more widespread in eukaryotic cells . Here we discuss the roles of a recently identified family of nucleotidyl transferases (encoded by the cid1 gene family) in cell cycle regulation . To date, this family has been characterised mainly in yeasts, but it is conserved throughout the eukaryotes . Biochemical studies have indicated that a subset of members of this family function as cytoplasmic poly(A) polymerases targeting specific mRNAs for translation . This form of translational control appears to be particularly important for cell cycle regulation following inhibition of DNA synthesis . Biol Pharm Bull, 2002 Oct, 25(10), 1307 - 10 Antifungal evaluation of bis Mannich bases derived from acetophenones and their corresponding piperidinols and stability studies; Gul HI et al.; The development of resistance to current antifungal therapeutics drives the search for effective new agents . The fact that some acetophenone-derived Mannich bases had shown antifungal activities in our previous studies led us to design and synthesize acetophenone-derived bis Mannich bases, B1-B5, bis(beta-aroylethyl)methylamine hydrochlorides, to evaluate their antifungal activity . These bis Mannich bases were then converted to the corresponding piperidinols, C1-C5, which are structural isomers of bis derivatives, 3-aroyl-4-aryl-1-methyl-4-piperidinol hydrochlorides, to see alterations in biological activity . A stability study of B1 and Cl was also carried out to estimate whether they alkylate the thiols . All compounds studied have shown antifungal activity, especially against dermatophytes (Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, and Microsporum canis), in the concentration range studied (2-128 microng/ml) . The activity was especially apparent against T . tonsurans . All compounds had at least equal antifungal activity compared with the reference compound amphotericin-B against T . tonsurans . Bis Mannich bases were generally found to be more potent compounds than their structural isomer piperidinols . The results of our stability studies suggest that thiol alkylation may contribute to the antifungal activity of the Mannich bases synthesized . Even though all compounds showed antifungal activity against dermatophytes, bis Mannich bases B1, B2, B4, and B5 appear to have potential for developing novel antifungal agents against dermatophytes. Int J Dermatol, 2002 Oct, 41(10), 647 - 51 Epidemiology of onychomycosis in patients with diabetes mellitus in India; Dogra S et al.; BACKGROUND: The number of individuals diagnosed with diabetes mellitus is increasing worldwide . Although onychomycosis is often observed in diabetics, there have been no large studies of its epidemiology in this patient group in India . METHODS: We studied the prevalence of onychomycosis in diabetics attending the Diabetes Clinic at the Postgraduate Institute of Medical Education and Research, Chandigarh, India, and compared it with that in a nondiabetic control group . A total of 400 diabetic subjects (237 males, 163 females), aged 48.8 +/- 0.5 years (mean +/- SD), were evaluated . RESULTS: The prevalence of onychomycosis in the diabetic and control groups was 17% and 6.8%, respectively, the difference being statistically significant (P < 0.001) . The presence of onychomycosis was found to correlate significantly with increasing age (P < 0.01) and male gender (P < 0.05) in both diabetic and control groups . From diabetics, yeasts were the most common isolate (48.1%), followed by dermatophytes and nondermatophyte molds in 37% and 14.8%, respectively . In the control group, the distribution of yeasts, dermatophytes, and nondermatophyte molds was 25%, 62.5%, and 12.5%, respectively . After controlling for age and sex, a stepwise logistic regression demonstrated that significant predictors for onychomycosis included the duration of diabetes (P < 0.01), absent or feeble peripheral pulses (P < 0.15), peripheral neuropathy (P < 0.05), and retinopathy (P < 0.001) . CONCLUSIONS: Diabetics were found to be 2.5 times more likely to have onychomycosis than the controls . Predisposing factors included increasing age, male gender, duration of diabetes, impaired peripheral circulation, peripheral neuropathy and retinopathy. Mol Biochem Parasitol, 2002 Sep-Oct, 124(1-2), 51 - 62 Peroxisomal targeting protein 14 (PEX14) from Leishmania donovani . Molecular, biochemical, and immunocytochemical characterization; Jardim A et al.; Pathogens of the Leishmania and Trypanosoma genera compartmentalize glycolytic and other nutritional pathways in glycosomes, unique subcellular organelles related to the peroxisomes of mammals and yeasts . Most glycosomal proteins are targeted to the glycosomes by a COOH-terminal tripeptide signal similar to the peroxisomal targeting signal-1 (PTS-1) . It has been proposed that PTS-1 forms a complex with the PEX5 receptor protein which then docks to the glycosomal membrane through interactions with the membrane associated PEX14 protein . To analyze the role of PEX14 in glycosomal protein import, the gene encoding the L . donovani PEX14 (LdPEX14) was isolated and shown to encode a 464 amino acid protein that exhibited very limited sequence homology with peroxisomal PEX14 proteins . In vitro binding experiments with purified recombinant LdPEX14 and LdPEX5 confirmed that LdPEX14-LdPEX5 interacted with a K(d) of 2.75 microM . When LdPEX5 was preloaded with a PTS-1 peptide, the affinity of the LdPEX14-LdPEX5 interaction affinity increased . Furthermore, binding experiments with truncated forms of LdPEX5 and LdPEX14 showed that the interaction domains localized to the amino terminal region of both proteins . Finally, confocal microscopy, subcellular fractionation, and differential extraction experiments indicated that LdPEX14 is a soluble protein that associates tightly with the glycosomal membrane and further support the role of LdPEX14 in forming a docking complex involved in glycosome biogenesis. Curr Opin Cell Biol, 2002 Aug, 14(4), 434 - 47 Phosphoinositides and the golgi complex; De Matteis M et al.; Phosphoinositides act as precursors of second messengers and membrane ligands for protein modules . Specific lipid kinases and phosphatases are located and differentially regulated in cell organelles, generating a non-uniform distribution of phosphoinositides . Although it is not clear whether and how the phosphoinositide pools are integrated, it is certain that they locally control fundamental processes, including membrane trafficking . This applies to the Golgi complex, where a direct, central role of the phosphatidylinositol 4,5-bisphosphate precursor phosphatidylinositol 4-phosphate has recently been reported. Plant Physiol, 2002 Oct, 130(2), 688 - 97 Arabidopsis ABI5 subfamily members have distinct DNA-binding and transcriptional activities; Kim SY et al.; A small family of novel basic leucine zipper proteins that includes abscisic acid (ABA)-INSENSITIVE 5 (ABI5) binds to the promoter region of the lea class gene Dc3 . The factors, referred to as AtDPBFs (Arabidopsis Dc3 promoter-binding factors), were isolated from an immature seed cDNA library . AtDPBFs bind to the embryo specification and ABA-responsive elements in the Dc3 promoter and are unique in that they can interact with cis-elements that do not contain the ACGT core sequence required for the binding of most other plant basic leucine zipper proteins . Analysis of full-length cDNAs showed that at least five different Dc3 promoter-binding factors are present in Arabidopsis seeds; one of these, AtDPBF-1, is identical to ABI5 . As expected, AtDPBF-1/ABI5 mRNA is inducible by exogenous ABA in seedlings . Despite the near identity in their basic domains, AtDPBFs are distinct in their DNA-binding, dimerization, and transcriptional activity. Genome Biol . 2002 Sep 24;3(10):REPORTS4032 . Epub 2002 Sep 24. Developmental biologists cast a net over sequenced genomes; Gerberding M et al.; A report on the annual meeting of the Society of Developmental Biology, Madison, Wisconsin, USA, 21-24 July 2002. J Biol Chem, 2002 Dec 20, 277(51), 49230 - 7 Epub 2002 Oct 04. Characterization of the two coactivator-interacting surfaces of the androgen receptor and their relative role in transcriptional control; Christiaens V et al.; The androgen receptor interacts with the p160 coactivators via two surfaces, one in the ligand binding domain and one in the amino-terminal domain . The ligand binding domain interacts with the nuclear receptor signature motifs, whereas the amino-terminal domain has a high affinity for a specific glutamine-rich region in the p160s . We here describe the implication of two conserved motifs in the latter interaction . The amino-terminal domain of the androgen receptor is a very strong activation domain constituent of Tau5, which is mainly active in the absence of the ligand binding domain, and Tau1, which is only active in the presence of the ligand binding domain . Both domains are, however, implicated in the recruitment of the p160s . Mutation analysis of the p160s has shown that the relative contribution of the two recruitment mechanisms via the signature motifs or via the glutamine-rich region depend on the nature of the enhancers tested . We propose, therefore, that the androgen receptor-coactivator complex has several alternative conformations, depending partially on the context of the enhancer. J Basic Microbiol, 2002, 42(5), 295 - 301 Laccase production by some Phlebia species; Arora DS et al.; The present study was carried out to examine the ability of four species of the genus Phlebia, viz . P . floridensis, P . brevispora, P . radiata and P . fascicularia, to produce the ligninolytic enzyme laccase in liquid culture . P . floridensis was the most active species that even surpassed laccase production by Trametes versicolor, and was chosen for further study . Among several carbon sources tested, malt extract turned out to be the best medium for subsequent laccase concentration by ammonium sulfate precipitation . Specific enzyme activity increased 12-fold during this procedure and a K(m) value of 0.33 mM was calculated for the resulting laccase preparation using guaiacol as the substrate . Concentrated laccase from P . floridensis was relatively thermostable and retained 70% and 15% of its activity after 1 h preincubation at 50 degrees C and 60 degrees C, respectively. Swiss Med Wkly, 2002 Jun 15, 132(23-24), 303 - 11 Antifungal chemotherapy: advances and perspectives; Groll AH et al.; Invasive fungal infections have emerged as important causes of morbidity and mortality in immunocompromised patients . In response to this challenge, the field of antifungal chemotherapy has considerably expanded . Fluconazole and itraconazole, introduced in the late 1980s, were the first durably useful alternatives to amphotericin B deoxycholate . The clinical development of the lipid formulations of amphotericin B, and, more recently, that of novel echinocandin derivatives and improved antifungal triazoles each represent milestones in antifungal drug research that have further amplified our therapeutic options . Major progress has been made in harmonising disease definitions, in defining the paradigms of antifungal intervention, and in designing and implementing clinical trials . Standardised methods for in vitro susceptibility testing of yeasts and filamentous fungi have become available, and pharmacodynamic concepts have entered preclinical and clinical drug development . This article reviews the evolution of therapeutic options over the past decade, advances in chemoprevention and empirical antifungal therapy, progress in early diagnosis and pre-emptive therapy, the promise of the new echinocandins and second generation triazoles, as well as perspectives for combination therapies and adjuvant immunoreconstitution . Invasive fungal infections will remain a frequent and important complication of modern medicine; the current momentum in the field of laboratory and clinical antifungal drug research provides hope for substantial progress in prevention and management of these life-threatening infections in the near future. Curr Biol, 2002 Oct 1, 12(19), 1670 - 4 Poleward microtubule flux is a major component of spindle dynamics and anaphase a in mitotic Drosophila embryos; Maddox P et al.; During cell division, eukaryotic cells assemble dynamic microtubule-based spindles to segregate replicated chromosomes . Rapid spindle microtubule turnover, likely derived from dynamic instability, has been documented in yeasts, plants and vertebrates . Less studied is concerted spindle microtubule poleward translocation (flux) coupled to depolymerization at spindle poles . Microtubule flux has been observed only in vertebrates, although there is indirect evidence for it in insect spermatocytes and higher plants . Here we use fluorescent speckle microscopy (FSM) to demonstrate that mitotic spindles of syncytial Drosophila embryos exhibit poleward microtubule flux, indicating that flux is a widely conserved property of spindles . By simultaneously imaging chromosomes (or kinetochores) and flux, we provide evidence that flux is the dominant mechanism driving chromosome-to-pole movement (anaphase A) in these spindles . At 18 degrees C and 24 degrees C, separated sister chromatids moved poleward at average rates (3.6 and 6.6 microm/min, respectively) slightly greater than the mean rates of poleward flux (3.2 and 5.2 microm/min, respectively) . However, at 24 degrees C the rate of kinetochore-to-pole movement varied from slower than to twice the mean rate of flux, suggesting that although flux is the dominant mechanism, kinetochore-associated microtubule depolymerization contributes to anaphase A. Proteins, 2002 Nov 15, 49(3), 302 - 20 Rebuilt 3D structure of the chloroplast f1 ATPase-tentoxin complex; Minoletti C et al.; The F1 part of the chloroplast H+ adenosine triphosphate (ATP)-synthase (CF1) strongly interacts with tentoxin, a natural fungous cyclic tetrapeptide known to inhibit the chloroplast enzyme and not the mammalian mitochondrial enzyme . Whereas the synthesis or the hydrolysis of ATP requires the stepwise rotation of the protein rotor gamma within the (alphabeta)3 crown, only one molecule of tentoxin is needed to fully inhibit the complex . With the help of an original homology modeling technique, based on robust distance geometry protocols, we built a tridimensional model of the alpha3beta3gamma CF1) subcomplex (3200 esidues), in which we introduced three different nucleotide occupancies to check their possible influence on the tentoxin binding site . Simultaneous comparison of three available high-resolution X-ray structures of F1, performed with a local structural alignment search tool, led to characterizing common structural blocks and the distorsions experienced by the complex during the catalytic turnover . The common structural blocks were used as a starting point of the spinach CF1 structure rebuilding . Finally, tentoxin was docked into its putative binding site of the reconstructed structure . The docking method was initially validated in the mitochondrial enzyme by its ability to relocate nucleotides into their original position in the crystal . Tentoxin binding was found possible to the two alpha/beta interfaces associated with the empty and adenosine diphosphate (ADP)-loaded catalytic sites, but not to the one associated with the ATP-loaded site . These results suggest a mechanism of CF1 inhibition by one molecule of tentoxin, by the impossibility of the alpha/beta interface bearing tentoxin to pass through the ATP-loaded state . Hum Mol Genet, 2002 Oct 1, 11(20), 2479 - 88 Cancer epigenomics; Plass C; Research in cancer epigenomics is driven by the development of novel technologies and the utilization of model organisms ranging from yeasts to plants to vertebrates . For decades, the search for cancer genes has focused on genetic defects that were used as tags for identification of these genes . With the realization that epigenetic modifications, most importantly DNA methylation events, are frequently involved in transcriptional changes in both tumor suppressor genes and oncogenes, techniques have been developed that support the identification of novel cancer genes altered by DNA methylation alone or in combination with genetic events . Recent data demonstrate that, in addition to DNA methylation, chromatin modifications are also involved in gene regulation . We are now beginning to understand this interesting interplay between chromatin modifications, DNA methylation and gene regulation . This review will summarize our current knowledge of DNA methylation and histone modification in normal cells, introduce emerging concepts that show the intimate link between DNA methylation and chromatin modifications, and highlight recent advancements in our understanding of aberrant DNA methylation, with special emphasis on genome-wide hypermethylation. Cell, 2002 Sep 6, 110(5), 545 - 50 RNA logic in time and space; Darnell RB; An EMBO-sponsored workshop entitled "Translational Control in Development and Neurobiology" was held recently in Mallorca, Spain . The talks covered mechanisms of translational regulation, particularly in terms of temporal and spatial regulation, over a wide range of biological systems. Med Mycol, 2002 Aug, 40(4), 383 - 6 High rate of vaginal infections caused by non-C . albicans Candida species among asymptomatic women; Dan M et al.; A prospective observational study of patients attending a gynecological clinic and those referred to a clinic for genitourinary infections was undertaken with the purpose of evaluating the relative prevalence of non-C . albicans Candida species among Candida isolates from the vagina in different clinical settings in an area with high occurrence of vulvovaginal candidiasis . The rate of non-C . albicans Candida species was 44.5% among asymptomatic women, 19.4% among those with sporadic vaginitis and 21% among patients with chronic vaginal symptoms (p < 0.001 for asymptomatic vs . pooled symptomatic women) . No increase in the rate of non-C . albicans Candida was observed during a period of 4 years (1995-1998) despite a 1.57-fold increase in the sales of azole antifungal agents . Unlike some previous reports we could not document an association of non-C . albicans Candida species with chronic vaginal symptoms or increased use of azole antifungal agents . The significantly higher rate of these yeasts in asymptomatic women is in accord with the known tendency of non-C . albicans Candida species to cause mild symptoms. Plant Physiol, 2002 Sep, 130(1), 179 - 89 Engineering herbicide metabolism in tobacco and Arabidopsis with CYP76B1, a cytochrome P450 enzyme from Jerusalem artichoke; Didierjean L et al.; The Jerusalem artichoke (Helianthus tuberosus) xenobiotic inducible cytochrome P450, CYP76B1, catalyzes rapid oxidative dealkylation of various phenylurea herbicides to yield nonphytotoxic metabolites . We have found that increased herbicide metabolism and tolerance can be achieved by ectopic constitutive expression of CYP76B1 in tobacco (Nicotiana tabacum) and Arabidopsis . Transformation with CYP76B1 conferred on tobacco and Arabidopsis a 20-fold increase in tolerance to linuron, a compound detoxified by a single dealkylation, and a 10-fold increase in tolerance to isoproturon or chlortoluron, which need successive catalytic steps for detoxification . Two constructs for expression of translational fusions of CYP76B1 with P450 reductase were prepared to test if they would yield even greater herbicide tolerance . Plants expressing these constructs had lower herbicide tolerance than CYP76B1 alone, which is apparently a consequence of reduced stability of the fusion proteins . In all cases, increased herbicide tolerance results from more extensive metabolism, as demonstrated with exogenously fed phenylurea . Beside increased herbicide tolerance, expression of CYP76B1 has no other visible phenotype in the transgenic plants . Our data indicate that CYP76B1 can function as a selectable marker for plant transformation, allowing efficient selection in vitro and in soil-grown plants . Plants expressing CYP76B1 may also be a potential tool for phytoremediation of contaminated sites. Plant Physiol, 2002 Sep, 130(1), 22 - 46 Inositol phospholipid metabolism in Arabidopsis . Characterized and putative isoforms of inositol phospholipid kinase and phosphoinositide-specific phospholipase C; Mueller-Roeber B et al.; Phosphoinositides (PIs) constitute a minor fraction of total cellular lipids in all eukaryotic cells . They fulfill many important functions through interaction with a wide range of cellular proteins . Members of distinct inositol lipid kinase families catalyze the synthesis of these phospholipids from phosphatidylinositol . The hydrolysis of PIs involves phosphatases and isoforms of PI-specific phospholipase C . Although our knowledge of the roles played by plant PIs is clearly limited at present, there is no doubt that they are involved in many physiological processes during plant growth and development . In this review, we concentrate on inositol lipid-metabolizing enzymes from the model plant Arabidopsis for which biochemical characterization data are available, namely the inositol lipid kinases and PI-specific phospholipase Cs . The biochemical properties and structure of characterized and genome-predicted isoforms are presented and compared with those of the animal enzymes to show that the plant enzymes have some features clearly unique to this kingdom. Mem Inst Oswaldo Cruz, 2002 Jul, 97(5), 747 - 50 Clinical and mycological study of scalp white piedra in the State of Paraíba, Brazil; Pontes ZB et al.; White piedra is a superficial mycoses characterized by nodules on the hair shaft, caused by the basidiomycetous yeasts . In the present study, clinical and mycological findings of scalp white piedra caused by Trichosporon spp . are related . Twenty three cases of scalp white piedra were observed with a high incidence in women (87%) and preschool children from 2 to 6 (74%) years old . These groups presented a relationship of dependence with this infection . Despite the low socio-economic status, poor standards of hygiene, (48% of the patients) as well as the fact that 30.4% of the children shared the same nursery, these factors were not significant for the transmission of the mycosis . These were the first reports of scalp white piedra in Joao Pessoa city, Paraiba, Brazil. J Mol Biol, 2002 Sep 6, 322(1), 123 - 35 Hydrophobic interactions at the Ccap position of the C-capping motif of alpha-helices; Ermolenko DN et al.; We investigated the possible role of residues at the Ccap position in an alpha-helix on protein stability . A set of 431 protein alpha-helices containing a C'-Gly from the Protein Data Bank (PDB) was analyzed, and the normalized frequencies for finding particular residues at the Ccap position, the average fraction of buried surface area, and the hydrogen bonding patterns of the Ccap residue side-chain were calculated . We found that on average the Ccap position is 70% buried and noted a significant correlation (R=0.8) between the relative burial of this residue and its hydrophobicity as defined by the Gibbs energy of transfer from octanol or cyclohexane to water . Ccap residues with polar side-chains are commonly involved in hydrogen bonding . The hydrogen bonding pattern is such that, the longer side-chains of Glu, Gln, Arg, Lys, His form hydrogen bonds with residues distal (>+/-4) in sequence, while the shorter side-chains of Asp, Asn, Ser, Thr exhibit hydrogen bonds with residues close in sequence (<+/-4), mainly involving backbone atoms . Experimentally we determined the thermodynamic propensities of residues at the Ccap position using the protein ubiquitin as a model system . We observed a large variation in the stability of the ubiquitin variants depending on the nature of the Ccap residue . Furthermore, the measured changes in stability of the ubiquitin variants correlate with the hydrophobicity of the Ccap residue . The experimental results, together with the statistical analysis of protein structures from the PDB, indicate that the key hydrophobic capping interactions between a helical residue (C3 or C4) and a residue outside the helix (C", C3' or C4') are frequently enhanced by the hydrophobic interactions with Ccap residues. Ultramicroscopy, 2002 Aug, 92(3-4), 243 - 50 Optical interference artifacts in contact atomic force microscopy images; Mendez-Vilas A et al.; Atomic force microscopy images are usually affected by different kinds of artifacts due to either the microscope design and operation mode or external environmental factors . Optical interferences between the laser light reflected off the top of the cantilever and the light scattered by the surface in the same direction is one of the most frequent sources of height artifact in contact (and occasionally non-contact) images . They are present when imaging highly reflective surfaces, or even when imaging non-reflective materials deposited onto reflective ones . In this study interference patterns have been obtained with a highly polished stainless steel planchet . The influence of these artifacts in surface roughness measurements is discussed, and a semi-quantitative method based on the fast Fourier transform technique is proposed to remove the artifacts from the images . This method improves the results obtained by applying the usual flattening routines. Nat Rev Genet, 2002 Sep, 3(9), 683 - 97 The art and design of genetic screens: filamentous fungi; Casselton L et al.; In the 1940s, screens for metabolic mutants of the filamentous fungus Neurospora crassa established the fundamental, one-to-one relationship between a gene and a specific protein, and also established fungi as important genetic organisms . Today, a wide range of filamentous species, which represents a billion years of evolutionary divergence, is used for experimental studies . The developmental complexity of these fungi sets them apart from unicellular yeasts, and allows the development of new screens that enable us to address biological questions that are relevant to all eukaryotes. Nat Rev Mol Cell Biol, 2002 Sep, 3(9), 639 - 50 Seven-transmembrane receptors; Pierce KL et al.; Seven-transmembrane receptors, which constitute the largest, most ubiquitous and most versatile family of membrane receptors, are also the most common target of therapeutic drugs . Recent findings indicate that the classical models of G-protein coupling and activation of second-messenger-generating enzymes do not fully explain their remarkably diverse biological actions. Plant J, 2002 Sep, 31(5), 663 - 73 The VERNALIZATION INDEPENDENCE 4 gene encodes a novel regulator of FLOWERING LOCUS C; Zhang H et al.; The late-flowering, vernalization-responsive habit of many Arabidopsis ecotypes is mediated predominantly through repression of the floral programme by the FLOWERING LOCUS C (FLC) gene . To better understand this repressive mechanism, we have taken a genetic approach to identify novel genes that positively regulate FLC expression . We identified recessive mutations in a gene designated VERNALIZATION INDEPENDENCE 4 (VIP4), that confer early flowering and loss of FLC expression in the absence of cold . We cloned the VIP4 gene and found that it encodes a highly hydrophilic protein with similarity to proteins from yeasts, Drosophila, and Caenorhabditis elegans . Consistent with a proposed role as a direct activator of FLC, VIP4 is expressed throughout the plant in a pattern similar to that of FLC . However, unlike FLC, VIP4 RNA expression is not down-regulated in vernalized plants, suggesting that VIP4 is probably not sufficient to activate FLC, and that VIP4 is probably not directly involved in a vernalization mechanism . Epistasis analysis suggests that VIP4 could act in a separate pathway from previously identified FLC regulators, including FRIGIDA and the autonomous flowering promotion pathway gene LUMINIDEPENDENS . Mutants lacking detectable VIP4 expression flower earlier than FLC null mutants, suggesting that VIP4 regulates flowering-time genes in addition to FLC . Floral morphology is also disrupted in vip4 mutants; thus, VIP4 has multiple roles in development. Plant J, 2002 Sep, 31(5), 565 - 76 Regulation of ADL6 activity by its associated molecular network; Lam BC et al.; Plant dynamin-like proteins consist of a group of high molecular weight GTPase with diverse structural arrangements and cellular localizations . In addition, unlike animal dynamins, there was no evidence for the involvement of any plant dynamin-like protein in clathrin-mediated vesicle trafficking . In this study we demonstrate that ADL6 (Arabidopsis dynamin-like protein 6), due to its domain arrangement, behaves similarly to the animal dynamins . The association of ADL6 with clathrin-coated vesicles was demonstrated by co-fractionation and immunocytochemical studies . ADL6 also interacted via its C-terminus with gamma-adaptin, an adaptor protein of clathrin-coated vesicles . Our results suggest that ADL6 participates in clathrin-mediated vesicle trafficking originating from the Golgi . In addition, our studies demonstrate that ADL6 intrinsic GTPase activity is regulated by its association with acidic phospholipids and an SH3 (Src homology 3)-containing protein. Angew Chem Int Ed Engl, 2002 Jul 2, 41(13), 2259 - 64 Chemical strategies for iron acquisition in plants; Staiger D; Iron is an essential element for plant nutrition . Although iron is the fourth most abundant element (3 %) of the earth's crust, it is not readily available because of its low solubility . Therefore, plants need an active mechanism to extract iron from the soil . They have evolved several chemical strategies to acquire iron ions and the physiology of these mechanisms has been known for a long time . Only recently, the use of molecular genetic approaches has led to a biochemical and molecular characterization of the players involved, thus providing an entry to the manipulation of iron uptake in plants. Anal Chem, 2002 Aug 15, 74(16), 4235 - 49 High-efficiency nanoscale liquid chromatography coupled on-line with mass spectrometry using nanoelectrospray ionization for proteomics; Shen Y et al.; We describe high-efficiency (peak capacities of approximately 10(3)) nanoscale (using column inner diameters down to 15 microm) liquid chromatography (nanoLC)/low flow rate electrospray (nanoESI) mass spectrometry (MS) for the sensitive analysis of complex global cellular protein enzymatic digests (i.e., proteomics) . Using a liquid slurry packing method with carefully selected packing solvents, 87-cm-length capillaries having inner diameters of 14.9-74.5 microm were successfully packed with 3-microm C18-bonded porous (300-A pores) silica particles at a pressure of 18,000 psi . With a mobile-phase delivery pressure of 10,000 psi, these packed capillaries provided mobile-phase flow rates as low as approximately 20 nL/min at LC linear velocities of approximately 0.2 cm/s, which is near optimal for separation efficiency . To maintain chromatographic efficiency, unions with internal channel diameters as small as 10 microm were specially produced for connecting packed capillaries to replaceable nanoESI emitters having orifice diameters of 2-10 microm (depending on the packed capillary dimensions) . Coupled on-line with a hybrid-quadrupole time-of-flight MS through the nanoESI interface, the nanoLC separations provided peak capacities of approximately 10(3) for proteome proteolytic polypeptide mixtures when a positive feedback switching valve was used for quantitatively introducing samples . Over a relatively large range of sample loadings (e.g., 5-100 ng, and 50-500 ng of cellular proteolytic peptides for 14.9- and 29.7-microm-i.d . packed capillaries, respectively), the nanoLC/nanoESI MS response for low-abundance components of the complex mixtures was found to increase linearly with sample loading . The nanoLC/nanoESI-MS sensitivity also increased linearly with decreasing flow rate (or approximately inversely proportional to the square of the capillary inner diameter) in the flow range of 20-400 nL/min . Thus, except at the lower loadings, decreasing the separation capillary inner diameter has an effect equivalent to increasing sample loading, which is important for sample-limited proteomic applications . No significant effects on recovery of eluting polypeptides were observed using porous C18 particles with surface pores of 300-A versus nonporous particles . Tandem MS analyses were also demonstrated using the high-efficiency nanoLC separations . Chromatographic elution time, MS response intensity, and mass measurement accuracy was examined between runs with a single column (with a single nanoESI emitter), between different columns (same and different inner diameters with different nanoESI emitters), and for different samples (various concentrations of cellular proteolytic peptides) and demonstrated robust and reproducible sensitive analyses for complex proteomic samples. Mol Cell, 2002 Aug, 10(2), 347 - 58 The three-dimensional structure of the autoproteolytic, nuclear pore-targeting domain of the human nucleoporin Nup98; Hodel AE et al.; Nup98 is a component of the nuclear pore that plays its primary role in the export of RNAs . Nup98 is expressed in two forms, derived from alternate mRNA splicing . Both forms are processed into two peptides through autoproteolysis mediated by the C-terminal domain of hNup98 . The three-dimensional structure of the C-terminal domain reveals a novel protein fold, and thus a new class of autocatalytic proteases . The structure further reveals that the suggested nucleoporin RNA binding motif is unlikely to bind to RNA . The C terminus also contains sequences that target hNup98 to the nuclear pore complex . Noncovalent interactions between the C-terminal domain and the cleaved peptide tail are visible and suggest a model for cleavage-dependent targeting of hNup98 to the nuclear pore. Curr Genet, 2002 Aug, 41(5), 311 - 22 Epub 2002 Jul 11. Maintenance of mitochondrial DNA integrity: repair and degradation; Kang D et al.; Mitochondria have their own genome, which is essential for proper oxidative phosphorylation and hence for a large part of ATP production in a cell . Although mitochondrial DNA-less (rho(0)) cells can survive under certain conditions, the integrity of the mitochondrial genome is critical for the survival of multicellular organisms . Mitochondrial DNA (mtDNA) is damaged more than nuclear DNA because mitochondria produce a large amount of reactive oxygen species and tend to accumulate toxic xenobiotics . Therefore, there is keen interest in mechanisms that maintain the integrity of mtDNA . DNA repair may play an important role . The repair of mtDNA has been investigated less intensely than nuclear DNA repair because, for a long time, it was thought that mitochondria lacked DNA repair systems . In fact, DNA damage can be repaired in mitochondria . Base-excision repair in mitochondria is well established . The enzymes responsible for mtDNA repair have been identified and are encoded by the same genes as their nuclear counterparts . Mitochondrion-targeting sequences are generated through alternative splicing of mRNAs, alternative use of transcription initiation sites, or alternative use of translation initiation sites . In addition to DNA repair, the degradation of damaged mtDNA may be tolerated because there are multiple copies of mtDNA molecules in a cell. Epidemiol Mikrobiol Imunol, 2002 Aug, 51(3), 131 - 4 {Resistance of microscopic fungi to selected antimycotics}; Kunova Z; The author attended to enquiries of developing resistance of microscopic fungi (yeasts, micromycetes) to antimycotics used in therapy and their mechanisms of action. Physiol Genomics, 2002 Aug 14, 10(2), 79 - 91 RNA interference of peroxisome-related genes in C . elegans: a new model for human peroxisomal disorders; Petriv OI et al.; RNA-mediated interference (RNAi) for the posttranscriptional silencing of genes was used to evaluate the importance of various peroxisomal enzymes and peroxins for the development of Caenorhabditis elegans and to compare the roles of these proteins in the nematode to their roles in yeasts and humans . The nematode counterparts of the human ATP-binding cassette half-transporters, the enzymes alkyldihydroxyacetonephosphate synthase and Delta(3,5)-Delta (2,4)-dienoyl-CoA isomerase, the receptors for peroxisomal membrane and matrix proteins (Pex19p and Pex5p), and components of the docking and translocation machineries for matrix proteins (Pex13p and Pex12p) are essential for the development of C . elegans . Unexpectedly, RNAi silencing of the acyl-CoA synthetase-mediated activation of fatty acids, the alpha- and beta-oxidation of fatty acids, the intraperoxisomal decomposition of hydrogen peroxide, and the peroxins Pex1p, Pex2p, and Pex6p had no apparent effect on C . elegans development . The described analysis of functional gene knockouts through RNAi provides a basis for the use of C . elegans as a valuable model system with which to study the molecular and physiological defects underlying the human peroxisomal disorders. Rev Argent Microbiol, 2002 Apr-Jun, 34(2), 95 - 9 Onychomycosis in João Pessoa City, Brazil; Pontes ZB et al.; Onychomycosis epidemiology is a combination of various factors which include, among others, clinical presentation, etiologic agents of the infection and the patient's history background . Out of a total of 672 nail samples examined, 460 (68.4%) were microscopy positive for fungi and 306 (66.5%) of these were culture positive, including Candida (82%), dermatophytes (13.4%), Trichosporon spp (3.6%) and nondermatophyte molds (1%) . Onychomycosis was more frequent in females (79.7%) than in males (20.3%) . These were more common in fingernails (96.1%) than in toenails (60%) and yeasts were the most isolated etiologic agents . Among the clinical presentations, paronychia (CP) (57.2%) and onycholysis (CO) (24.8%) were the most common, caused frequently by C . albicans in 52.6% and 60.5% of the cases, respectively . T . rubrum (44.4%) and Trichosporon spp (22.2%) were the most frequent species in the case of distal lateral subungual onychomycosis (DLSO) . Fusarium spp was the agent responsible for 33.3% of the cases of proximal subungual onychomycosis (PSO) and for 14.3% of white superficial onychomycosis (WSO), whereas Acremonium spp was responsible for 14.3% of the cases of WSO. Science, 2002 Sep 6, 297(5587), 1692 - 6 Epub 2002 Aug 08. Structure, mechanism, and regulation of the Neurospora plasma membrane H+-ATPase; Kuhlbrandt W et al.; Proton pumps in the plasma membrane of plants and yeasts maintain the intracellular pH and membrane potential . To gain insight into the molecular mechanisms of proton pumping, we built an atomic homology model of the proton pump based on the 2.6 angstrom x-ray structure of the related Ca2+ pump from rabbit sarcoplasmic reticulum . The model, when fitted to an 8 angstrom map of the Neurospora proton pump determined by electron microscopy, reveals the likely path of the proton through the membrane and shows that the nucleotide-binding domain rotates by approximately 70 degrees to deliver adenosine triphosphate (ATP) to the phosphorylation site . A synthetic peptide corresponding to the carboxyl-terminal regulatory domain stimulates ATPase activity, suggesting a mechanism for proton transport regulation. Am J Clin Pathol, 2002 Aug, 118(2), 278 - 86 Evaluation of the status of laboratory practices and the need for continuing education in medical mycology; Rosner ER et al.; A survey to determine the need for training in medical mycology was sent to 605 US laboratories . Training needs were determined by comparing actual laboratory mycology practices with recommended practices, documenting the extent of mycology training reported by employees, and asking respondents to specify the fungi they considered most difficult to identify . The response rate was 56.7% (with only 316 laboratories providing sufficient information) . Results showed a large degree of interlaboratory variation in practices and suggested that more judicious practices could lower costs and improve clinical relevance . Only 55.6% of laboratories reported that at least 1 employee attended a formal mycology continuing education program in the 4 years before the survey . Species of dermatophytes, dematiaceous fungi, and non-Candida yeasts were the most difficult to identify . Training may be needed in basic isolation procedures and in advanced topics such as identification of problematic molds and yeasts and antifungal susceptibility testing . Educators should consider clinical relevance and cost-containment without sacrificing quality when designing courses . Support for additional mycology training may improve if hospital and laboratory administrators are alerted to potential dangers and costs involved in treating patients with invasive fungal infections. Mol Cell Biochem, 2002 May-Jun, 234-235(1-2), 3 - 9 Cyclic oxidation and reduction of protein methionine residues is an important antioxidant mechanism; Stadtman ER et al.; Almost all forms of reactive oxygen species (ROS) oxidize methionine residues of proteins to a mixture of the R- and S-isomers of methionine sulfoxide . Because organisms contain methionine sulfoxide reductases (Msr's) that can catalyze the thioredoxin-dependent reduction of the sulfoxides back to methionine, it was proposed that the cyclic oxidation/reduction of methionine residues might serve as antioxidants to scavenge ROS, and also to facilitate the regulation of critical enzyme activities . We summarize here results of studies showing that organisms possess two different forms of Msr--namely, MsrA that catalyzes reduction of the S-isomer and MsrB that catalyzes the reduction of the R-isomer . Deletion of the msrA gene in mice leads to increased sensitivity to oxidative stress and to a decrease (40%) in the maximum lifespan . This suggests that elimination of both Msr's would have more serious consequences. Cell, 2002 Jul 12, 110(1), 13 - 8 Shmoos, rafts, and uropods- the many facets of cell polarity; Dustin ML; The recent Juan March Foundation meeting on "Regulation and functional insights in cellular polarity" focused on cellular polarity in yeasts, Dictyostelium, epithelial cells, fibroblasts, and immune cells . The molecular systems covered included membrane rafts, actin and tubulin cytoskeleton, polarized transcription, signaling, and cell-cell adhesion . Across these diverse biological and molecular systems, important general concepts emerged, including new ideas for establishing and maintaining polarity that are likely to be applicable across models and experimental systems. Mol Cell, 2002 Jul, 10(1), 105 - 15 90S pre-ribosomes include the 35S pre-rRNA, the U3 snoRNP, and 40S subunit processing factors but predominantly lack 60S synthesis factors; Grandi P et al.; We report the characterization of early pre-ribosomal particles . Twelve TAP-tagged components each showed nucleolar localization, sedimented at approximately 90S on sucrose gradients, and coprecipitated both the 35S pre-rRNA and the U3 snoRNA . Thirty-five non-ribosomal proteins were coprecipitated, including proteins associated with U3 (Nop56p, Nop58p, Sof1p, Rrp9, Dhr1p, Imp3p, Imp4p, and Mpp10p) and other factors required for 18S rRNA synthesis (Nop14p, Bms1p, and Krr1p) . Mutations in components of the 90S pre-ribosomes impaired 40S subunit assembly and export . Strikingly, few components of recently characterized pre-60S ribosomes were identified in the 90S pre-ribosomes . We conclude that the 40S synthesis machinery predominately associates with the 35S pre-rRNA factors, whereas factors required for 60S subunit synthesis largely bind later, showing an unexpected dichotomy in binding. J Clin Microbiol, 2002 Aug, 40(8), 2953 - 8 Multicenter comparative evaluation of six commercial systems and the national committee for clinical laboratory standards m27-a broth microdilution method for fluconazole susceptibility testing of Candida species; Morace G et al.; Fluconazole susceptibility among 800 clinical Candida isolates (60% C . albicans) and two control strains (C . krusei ATCC 6258 and C . parapsilosis ATCC 22019) was tested with the NCCLS M27-A method (gold standard) and six commercial products (Candifast, disk, Etest, Fungitest, Integral System Yeasts, and Sensititre YeastOne) . Results were classified as susceptible, susceptible-dose dependent, or resistant using M27-A breakpoints or, for Fungitest, Integral System Yeasts, and Candifast, as susceptible, intermediate, or resistant, according to the manufacturers' instructions . Concordance with NCCLS M27-A results was analyzed with the chi(2) test . Intra- and interlaboratory reproducibility was also evaluated . NCCLS M27-A (90.1%), Etest (93.1%), Sensititre YeastOne (93.1%), disk (96.7%), Fungitest (92.6%), Integral System Yeasts (40.6%), and Candifast (6.0%) classified the indicated percentages of C . albicans isolates as susceptible . Among non-C . albicans strains, the percentages of susceptible isolates were as follows: NCCLS M27-A, 74.0%; Etest, 83.8%; Sensititre YeastOne, 64.1%; disk, 60.6%; Fungitest, 76.6%; Integral System Yeasts, 28.3%; and Candifast, 27.4% . All methods except Candifast and Integral System Yeasts showed good agreement with NCCLS M27-A results for both C albicans and non-C . albicans isolates . Intralaboratory reproducibility was excellent for NCCLS M27-A, Etest, Sensititre YeastOne, disk, and Fungitest (88 to 91%) . Similar results emerged from the interlaboratory reproducibility evaluation . Our findings indicate that some commercial methods can be useful for fluconazole susceptibility testing of clinical Candida isolates . Those characterized by a lack of medium standardization and/or objective interpretative criteria should be avoided . Particular caution is necessary when testing is being done for clinical and epidemiological purposes. Mol Pathol, 2002 Aug, 55(4), 250 - 61 A role for CCN3 (NOV) in calcium signalling; Li CL et al.; AIMS: In animals and humans increased expression of CCN3 (NOV) is detected in tissues where calcium is a key regulator, such as the adrenal gland, central nervous system, bone and cartilage, heart muscle, and kidney . Because the multimodular structure of the CCN proteins strongly suggests that these cell growth regulators are metalloproteins, this study investigated the possible role of CCN3 in ion flux and transport during development, control of cell proliferation, differentiation, and pathobiology . METHODS: The isolation of CCN3 partners was performed by means of the two hybrid system . Yeasts were cotransfected with an HL60 cDNA library fused to the transactivation domain of the GAL4 transcription factor, and with a plasmid expressing CCN3 fused to the DNA binding domain of GAL4 . Screening of the recombinant clones selected on the basis of leucine, histidine, and tryptophan prototrophy was performed with a beta-galactosidase assay . After the interaction between CCN3 and its putative partners was checked with a GST (glutathione S-transferase) pull down assay, the positive clones were identified by cloning . To establish whether the CCN3 protein affected calcium ion flux, a dynamic imaging microscopy system was used, which allowed the fluorometric measurement of the intracellular calcium concentration . The proteins used in the assays were GST fused with either CCN3 or CCN2 (CTGF) and GST alone as a control . RESULTS: The two hybrid system identified the S100A4 (mts1) calcium binding protein as a partner of CCN3 and the use of the GST fusion proteins showed that the addition of CCN3 and CCN2 to G59 glioblastoma and SK-N-SH neuroblastoma cells caused a pronounced but transient increase of intracellular calcium, originating from both the entry of extracellular calcium and the mobilisation of intracellular stores . CONCLUSIONS: The interaction of CCN3 with S100A4 may account, in part, for the association of CCN3 with carcinogenesis and its pattern of expression in normal conditions . The increased intracellular calcium concentrations induced by CCN3 and CCN2 both involve different processes, among which voltage independent calcium channels might be of considerable importance in regulating the calcium flux associated with cell growth control, motility, and spreading . These observations assign for the first time a biological function to the CCN3 protein and point out a broader role for the CCN proteins in calcium ion signalling. Acta Biochim Pol, 2002, 49(1), 1 - 10 Aminoacyl-tRNA synthetases and aminoacylation of tRNA in the nucleus; Mucha P; This review is focused on findings concerning the presence of translation apparatus components (aminoacyl-tRNA synthetases, aminoacyl-tRNA, elongation factors) as well as translation itself in the nucleus . A nuclear role of these molecules is unknown . New findings suggest that well-accepted model of spatial segregation of transcription and translation in eukaryotic cell may be oversimplifcation . Nuclear coupling of both these processes show us how exciting and surprising may be the world of the living cell. Genetics, 2002 Jul, 161(3), 1089 - 99 Identification of six loci in which mutations partially restore peroxisome biogenesis and/or alleviate the metabolic defect of pex2 mutants in podospora; Ruprich-Robert G et al.; Peroxins (PEX) are proteins required for peroxisome biogenesis . Mutations in PEX genes cause lethal diseases in humans, metabolic defects in yeasts, and developmental disfunctions in plants and filamentous fungi . Here we describe the first large-scale screening for suppressors of a pex mutation . In Podospora anserina, pex2 mutants exhibit a metabolic defect {inability to grow on medium containing oleic acid (OA medium) as sole carbon source} and a developmental defect (inability to differentiate asci in homozygous crosses) . Sixty-three mutations able to restore growth of pex2 mutants on OA medium have been analyzed . They fall in six loci (suo1 to suo6) and act as dominant, allele-nonspecific suppressors . Most suo mutations have pleiotropic effects in a pex2(+) background: formation of unripe ascospores (all loci except suo5 and suo6), impaired growth on OA medium (all loci except suo4 and suo6), or sexual defects (suo4) . Using immunofluorescence and GFP staining, we show that peroxisome biogenesis is partially restored along with a low level of ascus differentiation in pex2 mutant strains carrying either the suo5 or the suo6 mutations . The data are discussed with respect to beta-oxidation of fatty acids, peroxisome biogenesis, and cell differentiation. Peptides, 2002 Jul, 23(7), 1351 - 9 Transmissible spongiform encephalopathies: the story of a pathogenic protein; Van Everbroeck B et al.; An overview is provided from the first description of the transmissible spongiform encephalopathies (TSE) to recent major discoveries in this research field . The TSE are a group of diseases in animal and in man caused by a unique pathogen: the prion protein . The exact nature of the etiological agent or the prion protein is thought to be a misfolded protein . Although current research has provided a wealth of data indicating that a structural isoform of the prion protein is the responsible pathogen, this hypothesis is not yet experimentally proven. Drug Resist Updat, 2002 Feb, 5(1), 3 - 10 Recent progress in the diagnosis of fungal infections in the immunocompromised host; Erjavec Z et al.; The management of invasive fungal infections has been hampered by the inability to diagnose the infection at an early stage of disease . Although proving the presence of infection by histology and culture remains the cornerstone of the diagnosis, non-culture based methods are becoming available that enable early detection . Molecular diagnosis by PCR appears very promising since fungal DNA can be detected in the blood of infected patients before conventional methods . Furthermore, a broad range of yeasts and molds can be identified to species level . Automation of sample preparation and use of real-time PCR systems will help standardize the procedure and reduce false positive results due to contamination . Promising assays for the detection of fungal antigens in serum have been commercialized, including detection systems for mannan (Candida) and galactomannan (Aspergillus) . Circulating antigens can be detected at an early stage of infection, often before the onset of clinical symptoms . Antigen detection is limited to detecting only one genus and not enabling speciation . Furthermore, both PCR and antigen detection can be used to monitor the response of patients to treatment with anti-fungal agents . Although prospective screening of high-risk patients for the presence of circulating markers of fungal infection appears to be an appropriate strategy, studies are needed to help to establish the optimal approach to managing invasive fungal infections that incorporates the benefits of non-culture based methods. Curr Opin Struct Biol, 2002 Jun, 12(3), 368 - 73 Computational methods for the prediction of protein interactions; Valencia A et al.; Establishing protein interaction networks is crucial for understanding cellular operations . Detailed knowledge of the 'interactome', the full network of protein-protein interactions, in model cellular systems should provide new insights into the structure and properties of these systems . Parallel to the first massive application of experimental techniques to the determination of protein interaction networks and protein complexes, the first computational methods, based on sequence and genomic information, have emerged. Trends Plant Sci, 2002 Jul, 7(7), 301 - 8 Mitogen-activated protein kinase cascades in plants: a new nomenclature; MAPK Group; Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules in eukaryotes, including yeasts, animals and plants . These protein phosphorylation cascades link extracellular stimuli to a wide range of cellular responses . In plants, MAPK cascades are involved in responses to various biotic and abiotic stresses, hormones, cell division and developmental processes . Completion of the Arabidopsis genome-sequencing project has revealed the existence of 20 MAPKs, 10 MAPK kinases and 60 MAPK kinase kinases . Here, we propose a simplified nomenclature for Arabidopsis MAPKs and MAPK kinases that might also serve as a basis for standard annotation of these gene families in all plants. J Biol Chem, 2002 Jul 19, 277(29), 26098 - 102 Epub 2002 May 06. A point mutation of the AF2 transactivation domain of the glucocorticoid receptor disrupts its interaction with steroid receptor coactivator 1; Kucera T et al.; Glucocorticoids cause a 10-fold increase in hepatic phosphoenolpyruvate carboxykinase (PEPCK) gene transcription through two low affinity glucocorticoid receptor (GR) binding sites and a complex array of accessory factor DNA elements and associated proteins . To analyze how co-activators interact with the GR in this context, we took advantage of the C656G GR mutant that binds ligand with very high affinity . This GR activates PEPCK gene transcription at a 500-fold lower dexamethasone concentration than does wild type GR . Transfected C656G GR containing additional mutations or deletions was tested on PEPCK gene expression in H4IIE hepatoma cells . We found that the AF2 domain is the only one of the three defined transactivation domains in GR that is required for PEPCK gene expression and that mutation of this domain disrupts the direct interaction of GR with steroid receptor coactivator 1 (SRC-1) . These data help define the functional interaction between GR and SRC-1 and further define the role of the GR in glucocorticoid-mediated expression of the PEPCK gene. Kidney Int, 2002 Aug, 62(2), 679 - 87 Efficient in vitro lowering of carbonyl stress by the glyoxalase system in conventional glucose peritoneal dialysis fluid; Inagi R et al.; BACKGROUND: Reactive carbonyl compounds (RCOs) present in heat-sterilized peritoneal dialysis (PD) fluid have been incriminated in the progressive deterioration of the peritoneal membrane observed in long-term PD patients . The present study utilized the glyoxalase I (GLO I) system as a new approach to lower in vitro the peritoneal fluid content of RCOs such as methylglyoxal (MGO), glyoxal (GO) and 3-deoxyglucosone (3-DG) . METHODS: GO, MGO, and 3-DG solutions or conventional glucose PD fluids were incubated in vitro with various RCO lowering compounds . The evolution of GO, MGO, and 3-DG levels was monitored by high-performance liquid chromatography . The tested compounds included aminoguanidine and glutathione (GSH), alone or together with GLO I . The human GLO I gene was overexpressed in Chinese hamster ovary (CHO) cells, or ubiquitously in transgenic mice . Cell supernatant of the CHO transfectant and protein extracts of various organs of the transgenic mice were also tested . RESULTS: Aminoguanidine incubated with MGO/GO/3-DG mixtures, promptly reduced RCO levels . GSH alone had a similar but milder and slower effect . Together with GLO I, it promptly decreased GO and MGO levels but was less efficient toward 3-DG . After incubation with glucose PD fluid, GSH together with GLO I had the same effect on MGO, GO, and 3-DG levels . Addition of transfected cell supernatant or tissue extracts overexpressing GLO I, together with GSH to either GO, MGO, or 3-DG solutions, promptly and markedly reduced GO and MGO but not 3-DG levels . CONCLUSIONS: GLO I together with GSH efficiently lowers glucose-derived RCOs, especially GO and MGO, both in conventional glucose PD fluids and in RCO solutions . The fact that genetically manipulated cells overexpressing GLO I activity have a similar effect suggests that maneuvers raising GLO I activity in peritoneal cells or in the peritoneal cavity might help prevent the deleterious effects of the peritoneal carbonyl stress in PD patients . The clinical relevance of this approach is yet to be documented. Int Immunopharmacol, 2002 May, 2(6), 797 - 806 Modulation of rat macrophage function by the Mangifera indica L . extracts Vimang and mangiferin; Garcia D et al.; Vimang is an aqueous extract of Mangiferia indica L., traditionally used in Cuba as an anti-inflammatory, analgesic and antioxidant . In the present study, we investigated the effects of Vimang and of mangiferin (a C-glucosylxanthone present in the extract) on rat macrophage functions including phagocytic activity and the respiratory burst . Both Vimang and mangiferin showed inhibitory effects on macrophage activity: (a) intraperitoneal doses of only 50-250 mg/kg markedly reduced the number of macrophages in peritoneal exudate following intraperitoneal injection of thioglycollate 5 days previously (though there was no significant effect on the proportion of macrophages in the peritoneal-exudate cell population); (b) in vitro concentrations of 0.1-100 microg/ml reduced the phagocytosis of yeasts cells by resident peritoneal and thioglycollate-elicited macrophages; (c) in vitro concentrations of 1-50 microg/ml reduced nitric oxide (NO) production by thioglycollate-elicited macrophages stimulated in vitro with lipopolysaccharide (LPS) and IFNgamma; and (d) in vitro concentrations of 1-50 microg/ml reduced the extracellular production of reactive oxygen species (ROS) by resident and thioglycollate-elicited macrophages stimulated in vitro with phorbol myristate acetate (PMA) . These results suggest that components of Vimang, including the polyphenol mangiferin, have depressor effects on the phagocytic and ROS production activities of rat macrophages and, thus, that they may be of value in the treatment of diseases of immunopathological origin characterized by the hyperactivation of phagocytic cells such as certain autoimmune disorders. Mol Endocrinol, 2002 Jul, 16(7), 1538 - 51 Evolution of glycoprotein hormone subunit genes in bilateral metazoa: identification of two novel human glycoprotein hormone subunit family genes, GPA2 and GPB5; Hsu SY et al.; The canonical members of the human glycoprotein hormone subunit family of cystine knot-forming polypeptides include the common alpha-subunit, and four beta-subunit genes, FSHbeta, LHbeta, TSHbeta, and hCGbeta . Using pairwise sequence analysis of the complete human genome, we have identified two novel glycoprotein hormone subunit-related genes . Based on unique sequence similarity to the alpha- and beta-subunits of glycoprotein hormones, they were named glycoprotein-alpha2 (GPA2) and glycoprotein-beta5 (GPB5), respectively . PCR analysis using a panel of human cDNAs from 14 different tissues demonstrated that GPB5 is similar to other beta-subunits showing restricted tissue expression, mainly in pituitary and brain . In contrast, the GPA2 transcript is found in diverse tissues . Furthermore, immunoreactive GPA2 and GPB5 were detected in the anterior pituitary of mouse and frog, whereas the expression of GPA2 and GPB5 in transfected cells resulted in the secretion of recombinant polypeptides in conditioned medium . After GenBank searches in lower organisms, glycoprotein hormone beta-subunit-related genes were identified from the genome of nematode Caenorhabditis elegans, hookworm Ancylostoma caninum, and Drosophila melanogaster . The evolutionary conservation of these invertebrate homologs can be seen in several key sequence characteristics, and the data suggest that the glycoprotein hormone beta-subunit gene ancestor evolved before the emergence of bilateral metazoa, thus providing a better understanding of the evolution of this group of classic polypeptide hormones and their receptors . Studies of the complete inventory of genes homologous to glycoprotein hormone subunits in the human genome and lower organisms will allow future functional characterization and identification of their respective receptors. Philos Trans R Soc Lond B Biol Sci, 2002 Jun 29, 357(1422), 749 - 60 Plant D-type cyclins and the control of G1 progression; Oakenfull EA et al.; The basic pattern of controls that operate during the G1 phase of the plant cell cycle shows much closer similarity to animals than to the yeasts and other fungi . The activity of D-type cyclin (CycD) kinases is induced in response to stimulatory signals, and these phosphorylate the plant homologue of the retinoblastoma tumour susceptibility (Rb) protein . It is likely that Rb phosphorylation results in the activation of genes under the control of E2F transcription factors, including those required for S phase entry . As the initial triggers of the cascade, attention has focused on the CycDs, and a family of 10 genes is present in Arabidopsis, divided into three major and three minor groups . Analysis to date suggests that these groups are functionally distinct. J Cell Sci, 2002 Jul 1, 115(Pt 13), 2651 - 8 Compartmentation of enzymes in a microbody, the glycosome, is essential in Trypanosoma brucei; Guerra-Giraldez C et al.; All kinetoplastids contain membrane-bound microbodies known as glycosomes, in which several metabolic pathways including part of glycolysis are compartmentalized . Peroxin 2 is essential for the import of many proteins into the microbodies of yeasts and mammals . The PEX2 gene of Trypanosoma brucei was identified and its expression was silenced by means of tetracycline-inducible RNA interference . Bloodstream-form trypanosomes, which rely exclusively on glycolysis for ATP generation, died rapidly upon PEX2 depletion . Insect-form (procyclic) trypanosomes do not rely solely on glycolysis for ATP synthesis . PEX2 depletion in procyclic forms resulted in relocation of most tested matrix proteins to the cytosol, and these mutants also died . Compartmentation of microbody enzymes is therefore essential for survival of bloodstream and procyclic T . brucei . In contrast, yeasts and cultured mammalian cells grow normally in the absence of peroxisomal membranes unless placed on selective media. Mycoses, 2002, 45 Suppl 1, 47 - 52 {Histologic studies on otomycosis}; Vennewald I et al.; Fungal infections of the ear are mostly described as mycoses of the auditory canal . The aim of our investigations was to find out how fungi colonize the ear in immunocompetent patients . In the years from 1993 to 2000, 128 patients suspected of having otomycosis were examined . Of these 115 patients suffered from chronic otitis media with persisting tympanum perforation and otorrhea . A further 13 patients had clinical signs of otitis externa only . In 54 out of 139 samples, fungi were found in the auditory canal, in five on the tympanic membrane, and in five in the middle ear . Two-thirds were isolated as moulds and one-third as yeasts . Dominating species were Aspergillus niger and Candida parapsilosis . Samples of 15 patients suspected of having mastoiditis or cholesteatoma were examined histologically . Fungal hyphae were observed in the middle ear cavity and/or between horny lamellae of cholesteatoma in 4 patients . In the middle ear of immunocompetent patients chronic-hyperplastic (polypous) inflammation was detected with increased production of mucus, which probably promotes the colonization with pathogenic fungi as in the middle ear just like in the auditory canal. Virchows Arch, 2002 Jun, 440(6), 573 - 82 Epub 2002 Apr 09. Telomere lengthening in telomerase-negative cells: the ends are coming together; Scheel C et al.; Telomeres are crucial for chromosomal stability and cell viability . Activation of telomerase, a specialized reverse transcriptase, is the predominant mechanism for maintaining telomere length and function in yeasts and human cells . Telomere maintenance is regarded as a key element in the immortalization of cells and hence in oncogenesis . Although more than 90% of all malignant tumors display telomerase activity, there appear to be alternatives to this mechanism . Alternative lengthening of telomeres (ALT) in the absence of telomerase has been described in various organisms and recently also in immortalized and transformed human cells . This article will discuss how ALT is detected and how it affects telomere morphology . It will review the frequency and relevance of ALT in in vitro immortalized cell lines, tumor-derived cell lines, and primary tumors . We have only begun to link mechanisms by means of which ALT may act in immortalized human cells to the growing knowledge about telomeres, and we can look forward to further fascinating insights into telomere biology . Our review will also emphasize recent advances in our understanding of the induction and repression of ALT and the demonstration of ALT in cancer cells in the light of new treatment strategies targeted against telomere maintenance mechanisms. Curr Opin Cell Biol, 2002 Jun, 14(3), 377 - 83 Replication timing and transcriptional control: beyond cause and effect; Gilbert DM; In general, transcriptionally active euchromatin replicates during the first half of S phase, whereas silent heterochromatin replicates during the second half . Moreover, changes in replication timing accompany key stages of development . Although there is not a strict correlation between replication timing and transcription per se, recent results reveal a strong relationship between heritably repressed chromatin and late replication that is conserved in all eukaryotes . A long-standing question is whether replication timing dictates the structure of chromatin or vice versa . Mounting evidence supports a model in which replication timing is both cause and consequence of chromatin structure by providing a means to inherit chromatin states that, in turn, regulate replication timing in the subsequent cell cycle . Moreover, new findings relating aberrations in replication timing to defects in centromere function, chromosome cohesion and genome instability suggest that the role of replication timing extends beyond its relationship to transcription . Novel systems in both yeasts and mammals are finally beginning to reveal some of the determinants that regulate replication timing, which should pave the way for a long-anticipated molecular dissection of this complex liaison. J Cell Biol, 2002 Jun 10, 157(6), 941 - 51 Epub 2002 Jun 10. Rlp7p is associated with 60S preribosomes, restricted to the granular component of the nucleolus, and required for pre-rRNA processing; Gadal O et al.; Many analyses have examined subnucleolar structures in eukaryotic cells, but the relationship between morphological structures, pre-rRNA processing, and ribosomal particle assembly has remained unclear . Using a visual assay for export of the 60S ribosomal subunit, we isolated a ts-lethal mutation, rix9-1, which causes nucleolar accumulation of an Rpl25p-eGFP reporter construct . The mutation results in a single amino acid substitution (F176S) in Rlp7p, an essential nucleolar protein related to ribosomal protein Rpl7p . The rix9-1 (rlp7-1) mutation blocks the late pre-RNA cleavage at site C2 in ITS2, which separates the precursors to the 5.8S and 25S rRNAs . Consistent with this, synthesis of the mature 5.8S and 25S rRNAs was blocked in the rlp7-1 strain at nonpermissive temperature, whereas 18S rRNA synthesis continued . Moreover, pre-rRNA containing ITS2 accumulates in the nucleolus of rix9-1 cells as revealed by in situ hybridization . Finally, tagged Rlp7p was shown to associate with a pre-60S particle, and fluorescence microscopy and immuno-EM localized Rlp7p to a subregion of the nucleolus, which could be the granular component (GC) . All together, these data suggest that pre-rRNA cleavage at site C2 specifically requires Rlp7p and occurs within pre-60S particles located in the GC region of the nucleolus. Bioorg Med Chem, 2002 Aug, 10(8), 2657 - 62 Non-thiol farnesyltransferase inhibitors: utilization of an aryl binding site by 5-arylacryloylaminobenzophenones; Mitsch A et al.; We recently described a novel aryl binding site of farnesyltransferase . The 2-naphthylacryloyl residue was developed as an appropriate substituent for our benzophenone-based AAX-peptidomimetic capable of occupying this binding site, resulting in a non-thiol farnesyltransferase inhibitor with nanomolar activity . The activity of this inhibitor is readily explained on the basis of docking studies which show the 2-naphthyl residue fitting into the aryl binding site. Structure (Camb), 2002 Jun, 10(6), 763 - 72 Intra- and intermolecular interactions in sucrose transporters at the plasma membrane detected by the split-ubiquitin system and functional assays; Reinders A et al.; Interaction of two separately expressed halves of sucrose transporter SUT1 was detected by an optimized split-ubiquitin system . The halves reconstitute sucrose transport activity at the plasma membrane with affinities similar to the intact protein . The halves do not function independently, and an intact central loop is not required for membrane insertion, plasma membrane targeting, and transport . Under native conditions, the halves associate into higher molecular mass complexes . Furthermore, the N-terminal half of the low-affinity SUT2 interacts functionally with the C-terminal half of SUT1 . Since the N terminus of SUT2 determines affinity for sucrose, the reconstituted chimera has lower affinity than SUT1 . The split-ubiquitin system efficiently detects intramolecular interactions in membrane proteins, and can be used to dissect transporter structure. Biochem Biophys Res Commun, 2002 May 31, 294(1), 145 - 8 Native transfer RNA catalyzes Diels-Alder reaction; Mielcarek M et al.; In this paper we show that transfer ribonucleic acids (tRNAs) catalyze the Diels-Alder cycloaddition reaction . A new DNA oxidative damage product, 6-furfuryladenine (kinetin) or its riboside (diene), was transformed with dimethyl acetylenedicarboxylate or maleic anhydride (dienophile) . The reaction proceeds in the presence of tRNA at high pressure but not at ambient condition . If so tRNA in prebiotic conditions (RNA world) had at least two functions: catalytic and a carrier of genetic information . It means that tRNA at high pressure shows catalytic properties and is a true Diels-Alderase. Int J Infect Dis, 2002 Mar, 6 Suppl 1, S47 - 53 Antifungal drug resistance: does it matter? Rogers TR. The objectives of this review are: to review the modes of action of currently available antifungal drugs; to define drug resistance and discuss the mechanisms by which fungi can develop resistance to antifungal drugs; to consider the epidemiological and host factors that contribute to the outcome of antifungal therapy and whether the available in vitro susceptibility test methods can reliably predict clinical response; and to assess the overall relevance of drug resistance to the outcome of fungal infections.The incidence of antifungal drug resistance among pathogens causing invasive fungal infections appears to be increasing . In the case of Candida spp., this may in part be a consequence of selective pressure brought about by more intensive antifungal use leading to a 'pathogen shift' . Non-albicans Candida spp . are more likely to demonstrate reduced susceptibility to fluconazole compared to C . albicans . Susceptibility breakpoints developed by the National Committee for Clinical Laboratory Standards to test azoles and flucytosine against Candida spp . are helpful in guiding therapy . Antifungal drug resistance in yeasts is of clinical importance . Increasingly reliable methods of in vitro susceptibility testing can help predict clinical response to therapy, but other considerations, including host- and drug-related factors, can also have an important bearing on the ultimate outcome of treatment. Mycopathologia, 2002, 154(1), 25 - 8 Edible dates (Phoenix dactylifera), a potential source of Cladosporium cladosporioides and Sporobolomyces roseus: implications for public health; Moore JE et al.; Edible dates (Phoenix dactylifera) were examined for the presence of endogenous yeasts and filimentous fungi . Mean counts of fungi were 530 colony forming units (cfu) per gram of fruit, representing a mixture of two phenotypic colony types . Subsequent DNA extraction and PCR amplification of these two morphotypes yielded an amplicon of approximately 350 bp with the 5.8S-28S rRNA ITS region . Sequence analysis identified these to be Cladosporium cladosporioides (230 cfu/g) and Sporobolomyces roseus . Both organisms have been previously reported in opportunistic infections involving skin or in immunocompromised patients . This is the first report of edible dates being a source of these organisms and we emphasize the importance of the common practice of washing hands following the consumption of these fruits by hand. J Biol Chem, 2002 Jul 19, 277(29), 26281 - 5 Epub 2002 May 14. Mediation of the DCC apoptotic signal by DIP13 alpha; Liu J et al.; DCC (deleted in colorectal cancer) is a candidate tumor suppressor gene . However the function of DCC remains elusive . Previously, we demonstrated that forced expression of DCC induces apoptosis or cell cycle arrest (Chen, Y . Q., Hsieh, J . T., Yao, F., Fang, B., Pong, R . C., Cipriano, S . C . & Krepulat, F . (1999) Oncogene 18, 2747-2754) . To delineate the DCC-induced apoptotic pathway, we have identified a protein, DIP13 alpha, which interacts with DCC . The DIP13 alpha protein has a pleckstrin homology domain and a phosphotyrosine binding domain . It interacts with a region on the DCC cytoplasmic domain that is required for the induction of apoptosis . Although ectopic expression of DIP13 alpha alone causes only a slight increase in apoptosis, co-expression of DCC and DIP13 alpha results in an approximately 5-fold increase in apoptosis . Removal of the DCC-interacting domain on DIP13 alpha abolishes its ability to enhance DCC-induced apoptosis . Inhibition of endogenous DIP13 alpha expression by small interfering RNA blocks DCC-induced apoptosis . Our data suggest that DIP13 alpha is a mediator of the DCC apoptotic pathway. Clin Microbiol Infect, 2002 Mar, 8(3), 162 - 73 Identification of Malassezia species from patient skin scales by PCR-RFLP; Gaitanis G et al.; OBJECTIVE: This study was aimed at the development of a DNA-based procedure directly applicable to pathological skin scales and at the assessment of its value in rapid laboratory confirmation and identification of each of the seven Malassezia species . These lipophilic basidiomycetous yeasts in predisposed individuals are involved in pityriasis versicolor, seborrheic dermatitis, blepharitis, folliculitis, atopic dermatitis and fungemia . Standard identification procedures to species level are available, but so far no system for direct detection and characterization of Malassezia species in clinical specimens is available . METHODS: Malassezia DNA was extracted from pathological skin scales by a modified hexadecyltrimethylammonium bromide (CTAB) method and amplified by single and nested polymerase chain reaction (PCR), assays using the general fungal ITS 1/4 and 3/4 primers for amplification of sequences from the Malassezia major ribosomal DNA complex . Restriction fragment length polymorphism (RFLP) analysis of PCR products was used in subsequent species identification . DNA extracted from culture-positive skin scales was also tested by PCR and the RFLP patterns obtained were analyzed . RESULTS: A total of 36 isolates were tested . Distinct pure culture and skin-scale ITS 3/4 HinfI and AluI restriction patterns differentially identified M . furfur, M . globosa, M . restricta, M . sympodialis, M . pachydermatis, M . obtusa and M . slooffiae . Malassezia DNA was extracted from pathological skin scales and RFLP identified solitary and multiple Malassezia species in the same specimen . Molecular identification was confirmed by cultures and biochemical tests . Concurrent detection and identification of Candida and Yarrowia species was also feasible from skin scales . CONCLUSION: The proposed method, described for the first time, could provide a sensitive and rapid detection and identification system for Malassezia species, which may be applied to epidemiological surveys and routine practice. Biochim Biophys Acta, 2002 May 20, 1597(1), 36 - 41 The distinctiveness of ATP:citrate lyase from Aspergillus nidulans; Adams IP et al.; ATP:citrate lyase (ACL), an important enzyme in lipid synthesis, has been purified from Aspergillus nidulans to a specific activity of 19.6 micromol min(-1) mg(-1), almost twice that of any other purified ACL and shown to be distinct from any previously purified ACL . The enzyme is a 371+/-31 kDa hexamer of 3 alpha, 3 beta proteins, unlike the 4 alpha tetramer found in rats or yeasts . The molecular weights of the alpha and beta protein subunits were determined by SDS-PAGE to be 70 and 55 kDa.ACL in A . nidulans (unlike Aspergillus niger) appears to be regulated by the carbon source present in the media . In crude extracts, it was found at high activity (88 micromol min(-1) mg protein(-1)) in glucose-grown cells but only at low activity (10 micromol min(-1) mg protein(-1)) in acetate-grown cells. Biochemistry, 2002 May 7, 41(18), 5765 - 75 Mapping the G-actin binding surface of cofilin using synchrotron protein footprinting; Guan JQ et al.; Cofilin is an actin regulatory protein that binds to both monomeric and filamentous actin, and has filament severing activity . Although crystal structures for the monomeric forms of both G-actin and cofilin have been described, the structure of the binary cofilin-G-actin complex is not available . Synchrotron protein footprinting is used to identify specific side chain residues on the cofilin surface that are buried in the formation of the cofilin-G-actin binary complex . Exposure to synchrotron X-rays results in stable oxidative modifications of aromatic, aliphatic, and sulfur-containing side chains, with the rate of modification for a particular residue being dependent on its intrinsic reactivity and solvent accessibility . The rates of modification were monitored for a number of peptides generated by digestion of oxidized cofilin, both in isolation and in its binary complex with G-actin . After binding to G-actin takes place, a significant decrease in modification rates, indicating protection of side chain groups, is seen for cofilin peptides corresponding to residues 4-20, 10-17, 83-96, 91-105, and 106-117 . A number of other peptides show no change in reactivity, and are presumed to represent regions distal to the binding site . Tandem mass spectrometry demonstrates that residues Leu 13, Pro 94, Met 99, and Leu 108 and 112 directly participate in the binding interface . These results are generally consistent with, and complementary to, the results of previous site-directed mutagenesis studies and extend our understanding of the G-actin binding surface of cofilin. Brain Res Mol Brain Res, 2002 Mar 28, 99(2), 145 - 9 Protein inhibitor of neuronal nitric oxide synthase interacts with protein kinase A inhibitors; Yu J et al.; Protein kinase A (PKA) and neuronal nitric oxide synthase (nNOS) are important signaling molecules . It is well known that PKA can specifically phosphorylate nNOS . But the underlying molecular mechanism is still obscure . Our data indicate that the protein inhibitor of nNOS (PIN) binds to protein kinase A inhibitors (PKIs), which suggests that PKIs, together with PIN, might mediate the phosphorylation of nNOS by PKA. Curr Opin Infect Dis, 2002 Apr, 15(2), 133 - 42 Malassezia species in skin diseases; Crespo Erchiga V et al.; Since the taxonomic revision carried out in 1996, enlarging the genus Malassezia to comprise seven different species, a number of studies have investigated from different points of view -- mycological, molecular and immunological -- the relationships of these species with the pathologies associated with lipophilic yeasts, as well as its presence in healthy skin . From these studies, it now appears clear that Malassezia globosa is the main species associated with pityriasis versicolor, which is the only cutaneous disease in which the involvement of Malassezia is undisputed . Nevertheless, this species can also be found in normal skin, in which the predominant species is Malassezia sympodialis . In the remaining dermatological disorders related to Malassezia, the role of these yeasts is controversial . In seborrhoeic dermatitis, atopic dermatitis and folliculitis, several studies have focused on the immunological aspects that could explain the pathogenic mechanism . In other diseases, such as confluent and reticulate papillomatosis, neonatal pustulosis, otitis and onychomycosis, the presence or significance of Malassezia is still a matter of dispute. Food Addit Contam, 2002 Apr, 19(4), 408 - 14 Potential ochratoxin A producers from wine grapes in Argentina and Brazil; Da RR et al.; The aim was to identify the normal mycoflora in wine grapes from Argentina and Brazil . We collected 50 grapes samples from Malbec and Chardonnay varieties in each country during the 1997-98 harvest . Yeasts were a major component of the fungal population, and the most frequent genera of filamentous fungi isolated were: Aspergillus, Penicillium and Botrytis . Other genera identified (in decreasing order) were: Phythophthora, Moniliella, Alternaria and Cladosporium . From grapes, the mean frequency of filamentous fungi ranged from 1.3 x 10(4) to 5.4 x 10(6) CFU g(-1) . We isolated 48 Aspergillus niger strains from Argentinian grape, of which eight could produce ochratoxin A . Sixteen of 53 A . niger strains from Brazilian grapes produced ochratoxin A . The results indicate that similar mycobiota were isolated from Argentinian and Brazilian wine grapes and there could be ochratoxin A production in this substrate. Proc Natl Acad Sci U S A, 2002 Apr 16, 99(8), 5331 - 6 The frameshift signal of HIV-1 involves a potential intramolecular triplex RNA structure; Dinman JD et al.; The cis-acting mRNA elements that promote programmed -1 ribosomal frameshifting present a natural target for the rational design of antiretroviral chemotherapies . It has been commonly accepted that the HIV-1 frameshifting signal is special, because its downstream enhancer element consists of a simple mRNA stem loop rather than a more complex secondary structure such as a pseudoknot . Here we present three lines of evidence, bioinformatic, structural, and genetic, showing that the biologically relevant HIV-1 frameshift signal contains a complex RNA structure that likely includes an extended RNA triple-helix region . We suggest that the potential intramolecular triplex structure is essential for viral propagation and viability, and that small molecules targeted to this RNA structure may possess antiretroviral activities. Mol Biol Cell, 2002 Apr, 13(4), 1390 - 407 Morphology and dynamics of the endocytic pathway in Dictyostelium discoideum; Neuhaus EM et al.; Dictyostelium discoideum is a genetically and biochemically tractable social amoeba belonging to the crown group of eukaryotes . It performs some of the tasks characteristic of a leukocyte such as chemotactic motility, macropinocytosis, and phagocytosis that are not performed by other model organisms or are difficult to study . D . discoideum is becoming a popular system to study molecular mechanisms of endocytosis, but the morphological characterization of the organelles along this pathway and the comparison with equivalent and/or different organelles in animal cells and yeasts were lagging . Herein, we used a combination of evanescent wave microscopy and electron microscopy of rapidly frozen samples to visualize primary endocytic vesicles, vesicular-tubular structures of the early and late endo-lysosomal system, such as multivesicular bodies, and the specialized secretory lysosomes . In addition, we present biochemical and morphological evidence for the existence of a micropinocytic pathway, which contributes to the uptake of membrane along side macropinocytosis, which is the major fluid phase uptake process . This complex endosomal compartment underwent continuous cycles of tubulation/vesiculation as well as homo- and heterotypic fusions, in a way reminiscent of mechanisms and structures documented in leukocytes . Finally, egestion of fluid phase from the secretory lysosomes was directly observed. Nat Cell Biol, 2002 May, 4(5), 329 - 36 The TRPM7 channel is inactivated by PIP(2) hydrolysis; Runnels LW et al.; TRPM7 (ChaK1, TRP-PLIK, LTRPC7) is a ubiquitous, calcium-permeant ion channel that is unique in being both an ion channel and a serine/threonine kinase . The kinase domain of TRPM7 directly associates with the C2 domain of phospholipase C (PLC) . Here, we show that in native cardiac cells and heterologous expression systems, G alpha q-linked receptors or tyrosine kinase receptors that activate PLC potently inhibit channel activity . Numerous experimental approaches demonstrated that phosphatidylinositol 4,5-bisphosphate (PIP(2)), the substrate of PLC, is a key regulator of TRPM7 . We conclude that receptor-mediated activation of PLC results in the hydrolysis of localized PIP(2), leading to inactivation of the TRPM7 channel. Mol Cell, 2002 Mar, 9(3), 505 - 14 A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids; Schwartz M et al.; We show that brome mosaic virus (BMV) RNA replication protein 1a, 2a polymerase, and a cis-acting replication signal recapitulate the functions of Gag, Pol, and RNA packaging signals in conventional retrovirus and foamy virus cores . Prior to RNA replication, 1a forms spherules budding into the endoplasmic reticulum membrane, sequestering viral positive-strand RNA templates in a nuclease-resistant, detergent-susceptible state . When expressed, 2a polymerase colocalizes in these spherules, which become the sites of viral RNA synthesis and retain negative-strand templates for positive-strand RNA synthesis . These results explain many features of replication by numerous positive strand RNA viruses and reveal that these viruses, reverse transcribing viruses, and dsRNA viruses share fundamental similarities in replication and may have common evolutionary origins. Mol Endocrinol, 2002 Apr, 16(4), 757 - 73 The nuclear receptor coactivators p300/CBP/cointegrator-associated protein (p/CIP) and transcription intermediary factor 2 (TIF2) differentially regulate PKA-stimulated transcriptional activity of steroidogenic factor 1; Borud B et al.; Steroidogenic factor-1 (SF-1) is a member of the nuclear receptor superfamily that plays essential roles in the development of endocrine organs . Steroid receptor coactivator 1 and transcription intermediary factor 2 (TIF2) belong to the p160 coactivator family that mediates transcriptional activation by several nuclear receptors, including SF-1 . Here, it is reported that another of the p160 coactivators, p/CIP, interacts with SF-1 through the activation function-2 domain . Both p300/CBP/cointegrator-associated protein (p/CIP) and TIF2 potentiated SF-1-mediated transcription from two reporter gene constructs in transfected nonsteroidogenic COS-1 cells and in adrenocortical Y1 cells . PKA was shown to stimulate SF-1 transcriptional activity, and coexpression of p/CIP together with the PKA catalytic subunit stimulated SF-1-mediated transactivation even further . In contrast, PKA catalytic subunit overexpression impaired the ability of TIF2 to potentiate SF-1-dependent transcription . Activation of PKA also inhibited the TIF2-mediated coactivation of other nuclear receptors such as PPAR alpha/-gamma and liver X receptor-alpha . The TIF2 mRNA levels were not affected by PKA, but instead we found that PKA activation led to a decrease in the levels of TIF2 protein . Moreover, the C-terminal activation domain 2 of TIF2 was required for the inhibitory effect of PKA, suggesting that this region is the target for the PKA-mediated down-regulation . Thus, in contrast to the regulation of p/CIP and steroid receptor coactivator 1, we suggest that activation of PKA leads to selective down-regulation of TIF2 and subsequently repression of TIF2 coactivator function. Biochem Biophys Res Commun, 2002 Mar 29, 292(2), 300 - 7 DOC-2/DAB2 is the binding partner of myosin VI; Inoue A et al.; Myosin VI is a molecular motor that moves processively along actin filaments and is believed to play a role in cargo movement in cells . Here we found that DOC-2/DAB2, a signaling molecule inhibiting the Ras cascade, binds to myosin VI at the globular tail domain . DOC-2/DAB2 binds stoichiometrically to myosin VI with one molecule per one myosin VI heavy chain . The C-terminal 122 amino acid residues of DOC-2/DAB2, containing the Grb2 binding site, is identified to be critical for the binding to myosin VI . Actin gliding assay revealed that the binding of DOC-2/DAB2 to myosin VI can support the actin filament gliding by myosin VI, suggesting that it can function as a myosin VI anchoring molecule . The C-terminal domain but not the N-terminal domain of DOC-2/DAB2 functions as a myosin VI anchoring site . The present findings suggest that myosin VI plays a role in transporting DOC-2/DAB2, a Ras cascade signaling molecule, thus involved in Ras signaling pathways . (c)2002 Elsevier Science (USA). Life Sci Space Res, 1972, 10, 211 - 25 An integrated multi-purpose biology instrument utilizing a single detector, the mass spectrometer; Radmer R et al.; A mass spectrometer is used to analyze the gas phase in a number of reaction vessels filled with Martian soil . By choosing appropriate incubation conditions this instrument can be used to perform a wide spectrum of experiments ranging from the observation of general indices of life, i.e . processes and patterns unexplainable by physico-chemical mechanisms, to assays utilizing isotopes which probe for specific metabolic processes . Of particular interest is the in situ incubation in which a Martian soil sample is maintained at a constant temperature and its gas phase composition analyzed with time . Properly interpreted, this is a very general life-detection probe which makes minimal assumption as to the nature of Martian biology . Other assays and measurements concerning the soil and the atmosphere compatible with this method are also described. Am J Clin Dermatol, 2002, 3(2), 71 - 81 Six novel antimycotics; Rubin AI et al.; We have reviewed six new antimycotic agents which have potential applications for human cutaneous and mucosal diseases . Information on these six drugs was obtained via an English language search of PubMed through the US National Library of Medicine . The antimycotic agents reviewed include rilopirox, lanoconazole, NND-502, butenafine, eberconazole and voriconazole . Rilopirox is a synthetic pyridone derivative, related to ciclopirox, with a fungicidal action . Rilopirox is a hydrophobic, topical agent with potential application in mucosal candida infections, tinea versicolor and seborrheic dermatitis . Lanoconazole, an imidazole, is a topical agent with potential application in tinea infections and cutaneous candidiasis . The drug has been available for clinical use in Japan since 1994 and once-daily application to affected areas is recommended . In addition to its antifungal effect, animal data suggest that application of lanoconazole 0.5 or 1% cream is associated with accelerated wound healing . NND-502, a stereoselective analog of lanoconazole, is a topical agent with potential application in tinea pedis infection . NND-502 appears to be more effective in inhibiting ergosterol biosynthesis than lanoconazole or bifonazole and clinical trials comparing these agents are awaited . Butenafine is the first member of a new class of antifungals, the benzylamine derivatives, and has been approved for topical use in Japan (since 1992) and the US . Butenafine has a potent fungicidal action and the drug has been shown to be effective in multiple clinical trials in patients with tinea pedis, tinea corporis and tinea cruris . Butenafine ha