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fungal growth inhibitory properties of new phytosphingolipid analogues.to study the growth inhibitory properties of a series of phytosphingosine (phs) and phytoceramide (phc) analogues.200818005346
a metabolome study of the steady-state relation between central metabolism, amino acid biosynthesis and penicillin production in penicillium chrysogenum.the relation between central metabolism and the penicillin biosynthesis pathway in penicillium chrysogenum was studied by manipulating the steady-state flux in both pathways. a high producing industrial strain was cultivated at a growth rate mu=0.05 h(-1) in glucose-limited chemostat cultures, both under penicillin-g producing and non-producing conditions. non-producing conditions were accomplished in two ways: (1) by cultivation without addition of the side chain precursor phenylacetic acid and ...200817905623
[preparation of a novel aminated fungal biomass and its sorption for typical endocrine disruptors].the fungal biomass of penicillium chrysogenum was used as raw materials to prepare the sorbent, and polyethylenimine (pei) was grafted onto the biomass surface in a two-step reaction. the best sorbent was obtained through the optimization experiments. as the pei-modified biomass was found to possess zero zeta-potential at ph 10.2, the biosorbent is favorable for the removal of anionic pentachlorophenol (pcp), 2,4-dichlorophenoxyacetic acid (2,4-d) and perfluorooctane sulphonate (pfos) potassium ...200818649533
rna-silencing in penicillium chrysogenum and acremonium chrysogenum: validation studies using beta-lactam genes expression.in this work we report the development and validation of a new rna interference vector (pjl43-rnai) containing a double-stranded rna expression cassette for gene silencing in the filamentous fungi penicillium chrysogenum and acremonium chrysogenum. classical targeted gene disruption in these fungi is very laborious and inefficient due to the low frequency of homologous recombination. the rnai vector has been validated by testing the attenuation of two different genes of the beta-lactam pathway; ...200818590779
post-translational enzyme modification by the phosphopantetheinyl transferase is required for lysine and penicillin biosynthesis but not for roquefortine or fatty acid formation in penicillium chrysogenum.nrpss (non-ribosomal peptide synthetases) and pkss (polyketide synthases) require post-translational phosphopantetheinylation to become active. this reaction is catalysed by a pptase (4'-phosphopantetheinyl transferase). the ppt gene of penicillium chrysogenum, encoding a protein that shares 50% similarity with the stand-alone large pptases, has been cloned. this gene is present as a single copy in the genome of the wild-type and high-penicillin-producing strains (containing multiple copies of t ...200818558918
phagocytosis of aspergillus fumigatus conidia by primary nasal epithelial cells in vitro.invasive aspergillosis, which is mainly caused by the fungus aspergillus fumigatus, is an increasing problem in immunocompromised patients. infection occurs by inhalation of airborne conidia, which are first encountered by airway epithelial cells. internalization of these conidia into the epithelial cells could serve as a portal of entry for this pathogenic fungus.200818564423
microbial and xanthine dehydrogenase inhibitory activity of some flavones.xanthine dehydrogenase (xdh) is responsible for the pathological condition called gout. in the present study different flavones synthesized from chalcone were evaluated in vitro for their inhibitory activity. inhibitory activity of flavones on xdh was determined in terms of inhibition of uric acid synthesis from xanthine. the enzymatic activity was found maximum at ph 7.5 and temperature 40 degrees c. the flavones 6-chloro-2-[3-(4-hydroxy-phenyl)-1-phenyl-1-h-pyrazol-4-yl]-chromen-4-one (f(1)) a ...200818569337
in vitro activity of penicillium chrysogenum antifungal protein (paf) and its combination with fluconazole against different dermatophytes.strains of five dermatophyte species (microsporum canis, microsporum gypseum, trichophyton mentagrophytes, trichophyton rubrum and trichophyton tonsurans) were selected for testing against penicillium chrysogenum antifungal protein (paf) and its combination with fluconazole (fcz). inhibition of microconidia germination and growth was detected with mics of paf ranging from 1.56 to 200 microg ml(-1) when it was used alone, or at constant concentration (100 microg ml(-1)) in combination with fcz at ...200818574706
isotopic non-stationary 13c gluconate tracer method for accurate determination of the pentose phosphate pathway split-ratio in penicillium chrysogenum.current (13)c labeling experiments for metabolic flux analysis (mfa) are mostly limited by either the requirement of isotopic steady state or the extremely high computational effort due to the size and complexity of large metabolic networks. the presented novel approach circumvents these limitations by applying the isotopic non-stationary approach to a local metabolic network. the procedure is demonstrated in a study of the pentose phosphate pathway (ppp) split-ratio of penicillium chrysogenum i ...200818555197
inactivation of conidia of penicillium chrysogenum, p. digitatum and p. italicum by ethanol solutions and vapours.a fractional factorial design, 2(5-1) experiments, was used for assessing the influence of 5 factors: water activity, aw [0.7, 0.9], temperature, t ( degrees c) [10, 30], mode of application, a [liquid, vapour], ethanol concentration, e (% w/w) [5, 10] and time, t (d) [1, 4] on the inactivation of spores of penicillium chrysogenum, p. digitatum and p. italicum. survival was determined by germination at optimal conditions within 3d. the experimental response was log (n 0/nt), where n 0 and nt (sp ...200818160164
antifungal activity stability of flaxseed protein extract using response surface methodology.the stability of the antifungal activity of flaxseed (linum usitatissimum) protein extract was evaluated in this study. response surface methodology (rsm) using box-behnken factorial design was used to evaluate the effects of treatment variables, that is, temperature (50 to 90 degrees c), time (1 to 29 min), and ph (2 to 8), on the residual antifungal activity (raa) against penicillium chrysogenum, fusarium graminearum, aspergillus flavus, and a penicillium sp. isolated from moldy noodles. regre ...200818211360
eighty years after its discovery, fleming's penicillium strain discloses the secret of its sex.eighty years ago, alexander fleming discovered antibacterial activity in the asexual mold penicillium, and the strain he studied later was replaced by an overproducing isolate still used for penicillin production today. using a heterologous pcr approach, we show that these strains are of opposite mating types and that both have retained transcriptionally expressed pheromone and pheromone receptor genes required for sexual reproduction. this discovery extends options for industrial strain improve ...200818223118
minimization of a eukaryotic mini-intein.inteins are internal protein splicing elements that can autocatalytically self-excise from their host protein and ligate the protein flanks (exteins) with a peptide bond. large inteins comprise independent protein splicing and endonuclease domains whereas mini-inteins lack the central endonuclease domain. to identify mini-intein domains that are essential for protein splicing, deletions were introduced at different sites of the 157-aa prp8 mini-intein of penicillium chrysogenum. the removal of e ...200818054328
fungistatic activity of flaxseed in potato dextrose agar and a fresh noodle system.although numerous researchers have studied flaxseed as a food ingredient for its health benefits, flaxseed (linum usitatissimum) has never been considered as a food preservative. the objective of this study was to investigate the effect of flaxseed flour (ff) concentration (0, 6, 9, 12, and 15% wt/wt), cultivar ('omega' and brown) and source (four seed companies located in minnesota and north dakota) on flaxseed fungistatic activity. fungal radial growth was used to assess the fungistatic activi ...200818077042
mold challenge study in bottled natural mineral waters and spring waters.microbiological challenge study was carried out to verify the microbial stability of bottled waters against four different mold species isolated from bottled water (fusarium sp.; cladosporium sp.; penicillium chrysogenum and aspergillus fumigatus) and to follow the growth of the molds in bottled water. twelve types of bottled water with different mineralization and co2 level in pet and glass packages were collected from 4 european countries. three different inoculation levels of spore suspension ...200818595319
mold species in dust from the international space station identified and quantified by mold-specific quantitative pcr.dust was collected over a period of several weeks in 2007 from hepa filters in the u.s. laboratory module of the international space station (iss). the dust was returned on the space shuttle atlantis, mixed, sieved and the dna was extracted. using a dna-based method called mold-specific quantitative pcr (msqpcr), 39 molds were measured in the dust. potential opportunistic pathogens aspergillus flavus and aspergillus niger and potential moderate toxin producers penicillium chrysogenum and penicil ...200818602989
expression of the acremonium chrysogenum ceft gene in penicillum chrysogenum indicates that it encodes an hydrophilic beta-lactam transporter.the acremonium chryrsogenum ceft gene encoding a membrane protein of the major facilitator superfamily implicated in the cephalosporin biosynthesis in a. chrysogenum was introduced into penicillium chrysogenum wisconsin 54-1255 (a benzylpenicillin producer), p. chrysogenum npe6 pyrg(-) (a derivative of wisconsin 54-1255 lacking a functional pende gene) and p. chrysogenum ta98 (a deacetylcephalosporin producer containing the cefd1, cefd2, cefef and cefg genes from a. chrysogenum). rt-pcr analysis ...200818668246
expression of the transporter encoded by the ceft gene of acremonium chrysogenum increases cephalosporin production in penicillium chrysogenum.by introduction of the cefef genes of acremonium chrysogenum and the cmch gene of streptomyces clavuligerus, penicillium chrysogenum can be reprogrammed to form adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid (ad7-accca), a carbamoylated derivate of adipoyl-7-aminodeacetoxy-cephalosporanic acid. the ceft gene of a. chrysogenum encodes a cephalosporin c transporter that belongs to the major facilitator superfamily. introduction of ceft into an ad7-accca-producing p. chrysogenum st ...200818691664
cloning and functional identification of c-4 methyl sterol oxidase genes from the penicillin-producing fungus penicillium chrysogenum.two c-4 methyl sterol oxidase genes (pcerg25a and pcerg25b) that are involved in ergosterol biosynthesis have been cloned from the penicillin-producing fungus penicillium chrysogenum. cdnas of both pcerg25a and pcerg25b have an orf 885 bp in length, encoding a peptide of 295 residues. the deduced amino acid sequences of pcerg25a and pcerg25b show 86% identity, and have high identities to the characterized c-4 methyl sterol oxidases from candida albicans and saccharomyces cerevisiae. the function ...200818707625
no-donors. part 17: synthesis and antimicrobial activity of novel ketoconazole-no-donor hybrid compounds.novel hybrid compounds combining the antifungal drug ketoconazole with a diazen-1-ium-1,2-diolate or an organic nitrate moiety and the corresponding no-donors without ketoconazole were synthesized and their activities against a broad variety of fungal strains were tested. hybridization modifies the spectrum of antimicrobial activities and generally, the ketoconazole-no-donor hybrids are more potent than ketoconazole. the no-donors alone show insufficient effectiveness.200818710813
antimicrobial effect of oxidized cellulose salts.antimicrobial properties of oxidized cellulose and its salts in linters (-l) and microsphere (-m) form (okcel h-l, okcel zn-m, okcel znna-l, okcel znna-m and okcel ag-l) were tested by a dilution method against a spectrum of microbial strains: escherichia coli, pseudomonas aeruginosa, staphylococcus epidermidis, bacillus licheniformis, aspergillus niger, penicillium chrysogenum, rhizopus oryzae, scopulariopsis brevicaulis, candida albicans and candida tropicalis. okcel ag-l exhibited antimicrobi ...200818712552
distributions of the growth rate of the germ tubes and germination time of penicillium chrysogenum conidia depend on water activity.the effects of water activities for sporulation (a(wsp)) and germination (a(wge)) on the distributions of the growth rate of the germ tubes (mu) and the germination time (t(g)) of penicillium chrysogenum conidia were determined by monitoring the length of the same germ tubes throughout the experiments automatically. no relationship between the individual t(g)'s and mu's could be established. irrespective of the water activity for germination, mu was greater and t(g) was less for conidia produced ...200818721680
lys89, lys90, and phe91 are critical core amino acid residues of the pen ch 18 major fungal allergen recognized by human ige antibodies.a vacuolar serine protease (pen ch 18) has been identified as a major allergen of penicillium chrysogenum. the molecular features of antigenic determinant(s) on pen ch 18 recognized by human ige antibodies, however, have remained unclear. here, we show that a dominant ige epitope on the n-terminally processed pen ch 18 allergen was narrowed down to residues 83-91. in addition, lys89, lys90, and possibly phe91 were identified as the core residues. substitution of lys89, lys90, or phe91 with alani ...200818760997
effect of the penicillium chrysogenum antifungal protein (paf) on barley powdery mildew and wheat leaf rust pathogens.the small molecular mass antifungal protein of penicillium chrysogenum (paf) inhibited the growths of two obligate biotrophic fungal pathogens, blumeria graminis f. sp. hordei and puccinia recondita f.sp. tritici and, hence, mitigated the symptoms of barley powdery mildew and wheat leaf rust infections, respectively. paf also affected adversely the germination of b. graminis conidia and p. recondita uredospores causing degenerative branching of germ tubes. since powdery mildews and rusts cause s ...200818798177
genome sequencing and analysis of the filamentous fungus penicillium chrysogenum.industrial penicillin production with the filamentous fungus penicillium chrysogenum is based on an unprecedented effort in microbial strain improvement. to gain more insight into penicillin synthesis, we sequenced the 32.19 mb genome of p. chrysogenum wisconsin54-1255 and identified numerous genes responsible for key steps in penicillin production. dna microarrays were used to compare the transcriptomes of the sequenced strain and a penicilling high-producing strain, grown in the presence and a ...200818820685
the heterotrimeric galpha protein pga1 regulates biosynthesis of penicillin, chrysogenin and roquefortine in penicillium chrysogenum.we have studied the role of the pga1 gene of penicillium chrysogenum, encoding the alpha subunit of a heterotrimeric g protein, in secondary metabolite production. the dominant activating pga1(g42r) mutation caused an increase in the production of the three secondary metabolites penicillin, the yellow pigment chrysogenin and the mycotoxin roquefortine, whereas the dominant inactivating pga1(g203r) allele and the deletion of the pga1 gene resulted in a decrease of the amount of produced penicilli ...200818957609
fungistatic activity of heat-treated flaxseed determined by response surface methodology.the objective of this study was to evaluate the effect of heat treatment on the fungistatic activity of flaxseed (linum usitatissimum) in potato dextrose agar (pda) medium and a fresh noodle system. the radial growth of penicilliumn chrysogenum, aspergillus flavus, and a penicillium sp. isolated from moldy noodles, as well as the mold count of fresh noodle enriched with heat treated flaxseed, were used to assess antifungal activity. a central composite design in the response surface methodology ...200819241553
the pattern of sensitisation to inhalant allergens in omani patients with asthma, allergic rhinitis and rhinoconjunctivitis.identification of relevant allergens that are prevalent in each environment which may have diagnostic and therapeutic implications in allergic diseases. this study aimed to identify the pattern of sensitisation to inhalant allergens in omani patients with asthma, allergic rhinitis and rhinoconjunctivitis.200821748078
ecological and physiological studies on soil fungi at western region, libya.sixty three species and 5 varieties belonging to 30 fungal genera were collected from 75 soil samples. cultivated (29 genera and 58 species + 5 var.), desert (22 and 35 + 2 var.) and saline soil (21 and 41 + 1 var.) fungi were recovered on glucose-, cellulose- and 50% sucrose-czapek's agar at 28℃. the most common genera were alternaria, aspergillus, emericella, fusarium, mycosphaerella, nectria and penicillium. the most prevalent species from the three types of soils on the three types of media ...200823997599
effect of some food preservatives on the lipolytic activity of beef luncheon fungi.beef luncheon meat is one of the most popular meals in several countries in the world including egypt. thirty one fungal species and 3 species varieties were recovered from 30 samples of beef luncheon meat collected from different supermarkets in qena. alternaria, aspergillus, emericella, mucor, mycosphaerella, penicillium and rhizopus were the most common genera on the two types of media. from the above genera, the most prevalent species were alternaria alternate, aspergillus flavus, a. fumigat ...200823997619
screening and productivity of penicillin antibiotic from penicillium sp.this paper highlights the antagonism effect of penicillium isolates, which were screened against the test organisms such as staphylococcus aureus, e. coli and penicillium sp. penicillium notatum and penicillium chrysogenum isolates were used for penicillin biosynthesis. the antibacterial activities of fermented crude penicillin extract were assayed by disc diffusion method. maximum antibacterial activity was observed in gram positive organisms (staphylococcus aureus) when compared with gram nega ...200921117415
diversity and pharmaceutical screening of fungi from benthic mats of antarctic lakes.during the micromat project, the fungal diversity of microbial mats growing in the benthic environment of antarctic lakes was accessed for the discovery of novel antibiotics and anticancers. in all, 160 filamentous fungi belonging to fifteen different genera and 171 yeasts were isolated from 11 lakes, classified and cultivated in different media and at different temperatures. filamentous fungi were then screened to discover novel antimicrobial and cytotoxic compounds. a total of 1422 extracts we ...200921798171
an efficient method for 15n-labeling of chitin in fungi.to permit facile (15)n solid-state nmr (ssnmr) analysis of the degree of acetylation (da) of chitinous materials in fungi a method for the introduction of a (15)n isotopic label has been developed. using penicillium chrysogenum as a model system, a series of (15)n-based media were surveyed for their abilities to support mycelial growth, and a rich medium supplemented with ((15)nh(4))(2)so(4) supported good growth. uptake of label into chitin extracted from mycelia grown in the rich ((15)nh(4))(2 ...200919249848
simultaneous quantification of free nucleotides in complex biological samples using ion pair reversed phase liquid chromatography isotope dilution tandem mass spectrometry.a new sensitive and accurate analytical method has been developed for quantification of intracellular nucleotides in complex biological samples from cultured cells of different microorganisms such as saccharomyces cerevisiae, escherichia coli, and penicillium chrysogenum. this method is based on ion pair reversed phase liquid chromatography electrospray ionization isotope dilution tandem mass spectrometry (ip-lc-esi-id-ms/ms. a good separation and low detection limits were observed for these com ...200919250917
isolation of lysozyme and an antifungal peptide from sea lamprey (petromyzon marinus) plasma.the sea lamprey (petromyzon marinus) belongs to the most primitive class of fish and has only innate immunity. the innate immune factors, lysozyme and an antifungal peptide, were isolated from sea lamprey plasma. sea lamprey plasma (40.1mg protein/ml) was assayed for lysozyme activity by gel diffusion assay. using hen egg white lysozyme standards, plasma concentration of lamprey lysozyme was 5microg lysozyme/mg total protein. the presence of lysozyme in such high concentration in lamprey plasma ...200919552965
using human sera to identify a 52-kda exoantigen of penicillium chrysogenum and implications of polyphasic taxonomy of anamorphic ascomycetes in the study of antigenic proteins.we are interested in isolating and identifying antigenic fungal proteins from species that grow on damp building materials. the indoor clade of penicillium chrysogenum, the so-called fleming clade, is the most common species of penicillium on moldy building materials. we have identified a 52-kda marker protein for the indoor clade of p. chrysogenum not present in a taxonomically diverse selection of fungi. it is found in high concentrations in protein extracted from the fungus grown on paper-fac ...200919590977
deletion and overexpression of the aspergillus nidulans gata factor areb reveals unexpected pleiotropy.the aspergillus nidulans transcription factor area is a key regulator of nitrogen metabolic gene expression. area contains a c-terminal gata zinc finger dna-binding domain and activates expression of genes necessary for nitrogen acquisition. previous studies identified areb as a potential negative regulator of nitrogen catabolism showing similarity with penicillium chrysogenum nreb and neurospora crassa asd4. the areb gene encodes multiple products containing an n-terminal gata zinc finger and a ...200919628561
construction of an hdfa penicillium chrysogenum strain impaired in non-homologous end-joining and analysis of its potential for functional analysis studies.the homologous recombination mechanism for dna-repair is not predominant in most filamentous fungi, resulting in extremely low targeting efficiencies for molecular engineering. to increase the gene targeting efficiency, it is becoming common practice to inactivate the non-homologous end-joining (nhej) pathway that causes random integration, by deleting the fungal homologs of the human ku70 and ku80 genes that encode proteins functioning in the nhej pathway. this has been described for several fi ...200919269344
engineering of penicillium chrysogenum for fermentative production of a novel carbamoylated cephem antibiotic precursor.penicillium chrysogenum was successfully engineered to produce a novel carbamoylated cephalosporin that can be used as a synthon for semi-synthetic cephalosporins. to this end, genes for acremonium chrysogenum expandase/hydroxylase and streptomyces clavuligerus carbamoyltransferase were expressed in a penicilling high-producing strain of p.chrysogenum. growth of the engineered strain in the presence of adipic acid resulted in production of adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxyl ...200919271269
pennicitrinone d, a new citrinin dimer from the halotolerant fungus penicillium notatum b-52. 200919282879
determination of microbial volatile organic compounds (mvocs) by passive sampling onto charcoal sorbents.passive sampling combined with gc-ms was applied for the detection of microbial volatile organic compounds (mvocs) emitted by mould cultures. the fungi species penicillium expansum, penicillium chrysogenum, penicillium brevicompactum, aspergillus fumigatus, aspergillus versicolor, aspergillus niger, and cladosporium cladosporoides were cultivated on dichloran glycerol agar. the emitted volatiles were collected by charcoal-based passive diffusion monitors during the first four weeks of growth. se ...200919289243
comparison of dry- and wet-based fine bead homogenizations to extract dna from fungal spores.the present study explored dna extraction kinetics from fungal spores, i.e., aspergillus niger, penicillium chrysogenum and cladosporium sphaerospermum, by fine bead mill homogenization. in particular, the study aimed to investigate basic differences between the dry- and wet-based methods. the results showed higher initial rates of the dna extractions by the dry-based method than by the wet-based method, due to higher collision efficiency among fine beads and fungal spores. based on the experime ...200919332310
biocontrol of meloidogyne incognita on tomato using antagonistic fungi, plant-growth-promoting rhizobacteria and cattle manure.biocontrol achieved by a single biocontrol agent is generally inconsistent under field conditions. the aim of the present study was to increase the competitiveness and efficacy of biocontrol agents by using them together with cattle manure.200919431151
functional aspects of the solution structure and dynamics of paf--a highly-stable antifungal protein from penicillium chrysogenum.penicillium antifungal protein (paf) is a promising antimycotic without toxic effects on mammalian cells and therefore may represent a drug candidate against the often lethal aspergillus infections that occur in humans. the pathogenesis of paf on sensitive fungi involves g-protein coupled signalling followed by apoptosis. in the present study, the solution structure of this small, cationic, antifungal protein from penicillium chrysogenum is determined by nmr. we demonstrate that paf belongs to t ...200919459942
molecular characterization of a fungal gene paralogue of the penicillin pende gene of penicillium chrysogenum.penicillium chrysogenum converts isopenicillin n (ipn) into hydrophobic penicillins by means of the peroxisomal ipn acyltransferase (iat), which is encoded by the pende gene. in silico analysis of the p. chrysogenum genome revealed the presence of a gene, pc13g09140, initially described as paralogue of the iat-encoding pende gene. we have termed this gene ial because it encodes a protein with high similarity to iat (ial for iat-like). we have conducted an investigation to characterize the ial ge ...200919470155
indoor fungal contamination of moisture-damaged and allergic patient housing analysed using real-time pcr.the aim of our study was to compare, using real-time (rt) pcr, quantitative levels of five fungal species in three kinds of dwellings.200919486284
quantitative pcr study on the mode of action of oligosaccharide elicitors on penicillin g production by penicillium chrysogenum.to investigate the effects of single and multiple additions of the oligosaccharide elicitors, obtained from alginate and locust bean gum, on penicillin g production and the transcript level of penicillin g biosynthetic genes.200919486407
application of raisin extracts as preservatives in liquid bread and bread systems.interest in natural ingredients with multifunctions in food has led to the investigation of the antimicrobial activity of raisins, a traditional baking ingredient. water and 60% ethanol extracts, and raisin juice concentrate (rjc, a commercial food ingredient) were assessed for their ability to inhibit the growth of ropy-bread-causing bacillus species. the ability of these additives and raisin paste were tested for antifungal activity in liquid bread model system and bread. in the liquid bread m ...200919490336
the global regulator laea controls penicillin biosynthesis, pigmentation and sporulation, but not roquefortine c synthesis in penicillium chrysogenum.the biosynthesis of the beta-lactam antibiotic penicillin is an excellent model for the study of secondary metabolites produced by filamentous fungi due to the good background knowledge on the biochemistry and molecular genetics of the beta-lactam producing microorganisms. the three genes (pcbab, pcbc, pende) encoding enzymes of the penicillin pathway in penicillium chrysogenum are clustered, but no penicillin pathway-specific regulators have been found in the genome region that contains the pen ...200918952140
heterologous production of non-ribosomal peptide lld-acv in saccharomyces cerevisiae.non-ribosomal peptides (nrps) are a diverse family of secondary metabolites with a broad range of biological activities. we started to develop an eukaryotic microbial platform based on the yeast saccharomyces cerevisiae for heterologous production of nrps using delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine (acv) as a model nrp. the penicillium chrysogenum gene pcbab encoding acv synthetase was expressed in s. cerevisiae from a high-copy plasmid together with phosphopantetheinyl transferase (p ...200919686863
comparison of skin prick tests with specific serum immunoglobulin e in the diagnosis of fungal sensitization in patients with severe asthma.it has been shown that patients with allergic bronchopulmonary aspergillosis (abpa) and patients with severe asthma with fungal sensitization (safs) can benefit from antifungal therapy. it is not known whether allergy skin prick tests (spt) or specific ige tests are more sensitive in the identification of patients who are sensitized to fungi and who are therefore candidates for antifungal therapy.200919689458
indoor mold concentration in eastern france.our prospective case-control study of 118 dwellings in eastern france examined fungal contamination in unhealthy dwellings (n = 32) (homes with visible mold contamination and adverse health outcomes reported by the occupants), dwellings occupied by allergic patients (with medical diagnostic and positive prick-tests for molds) (n = 27) and matched control dwellings (n = 59). unhealthy dwellings present higher airborne concentrations of aspergillus, penicillium, and cladosporium than control dwell ...200919840143
molds, mycotoxins, and sick building syndrome.the following is a review of some of the work we have done since 2004 regarding the importance of molds and their mycotoxins in the phenomenon of sick building syndrome (sbs). in these studies we showed that the macrocyclic trichothecene mycotoxins (mtm) of stachybotrys chartarum (sc) are easily dissociated from the surface of the organism as it grows and could therefore be consequently spread in buildings as the fungus experiences additional water events. we then showed that sc and penicillium ...200919854820
fungal spores: a critical review of the toxicological and epidemiological evidence as a basis for occupational exposure limit setting.fungal spores are ubiquitous in the environment. however, exposure levels in workplaces where mouldy materials are handled are much higher than in common indoor and outdoor environments. spores of all tested species induced inflammation in experimental studies. the response to mycotoxin-producing and pathogenic species was much stronger. in animal studies, nonallergic responses dominated after a single dose. allergic responses also occurred, especially to mycotoxin-producing and pathogenic speci ...200919863384
aspects on evolution of fungal beta-lactam biosynthesis gene clusters and recruitment of trans-acting factors.penicillins and cephalosporins are beta-lactam antibiotics. the formation of hydrophobic penicillins has been reported in fungi only, notably penicillium chrysogenum and aspergillus (emericella) nidulans, whereas the hydrophilic cephalosporins are produced by both fungi, e.g., acremonium chrysogenum (cephalosporin c), and bacteria. the producing bacteria include gram-negatives and gram-positives, e.g., streptomyces clavuligerus (cephamycin c) and lysobacter lactamgenus (cephabacins), respectivel ...200919863978
transcriptional upregulation of four genes of the lysine biosynthetic pathway by homocitrate accumulation in penicillium chrysogenum: homocitrate as a sensor of lysine-pathway distress.the lysine biosynthetic pathway has to supply large amounts of alpha-aminoadipic acid for penicillin biosynthesis in penicillium chrysogenum. in this study, we have characterized the p. chrysogenum l2 mutant, a lysine auxotroph that shows highly increased expression of several lysine biosynthesis genes (lys1, lys2, lys3, lys7). the l2 mutant was found to be deficient in homoaconitase activity since it was complemented by the aspergillus nidulans lysf gene. we have cloned a gene (named lys3) that ...200919696106
chapter 5: zones of inhibition? the transfer of information relating to penicillin in europe during world war ii.alexander fleming published his first description of penicillin in 1929, but the journal articles that were to propel penicillin from its relative obscurity were those of howard florey and his co-workers at oxford university. these were published in the lancet in the early years of world war ii and although wartime conditions restricted the flow of information on penicillin throughout europe, they never succeeded in shutting it off altogether. in germany an information-gathering initiative was e ...200919729093
preparation of fungal conidia impacts their susceptibility to inactivation by ethanol vapours.a common protocol employed for the preparation of conidia employs flooding a fungal colony grown on semi-solid media under optimum conditions with an aqueous solution. in contrast, conidia produced in a natural environment are usually not hydrated when disseminated in air and can be produced under water stress. in order to simulate the latter conditions, cultures were grown at different water activities and conidia were dry-harvested on the lid by turning the dishes upside-down then gently tappi ...200919762103
structure of the two-domain hexameric aps kinase from thiobacillus denitrificans: structural basis for the absence of atp sulfurylase activity.the tbd_0210 gene of the chemolithotrophic bacterium thiobacillus denitrificans is annotated to encode a 60.5 kda bifunctional enzyme with atp sulfurylase and aps kinase activity. this putative bifunctional enzyme was cloned, expressed and structurally characterized. the 2.95 a resolution x-ray crystal structure reported here revealed a hexameric assembly with d(3) symmetry. each subunit contains a large n-terminal sulfurylase-like domain and a c-terminal aps kinase domain reminiscent of the two ...200919770499
effect of a heterotrimeric g protein alpha subunit on conidia germination, stress response, and roquefortine c production in penicillium roqueforti.heterotrimeric g protein signaling regulates many processes in fungi, such as development, pathogenicity, and secondary metabolite biosynthesis. for example, the galpha subunit pga1 from penicillium chrysogenum regulates conidiation and secondary metabolite production in this fungus. the dominant activating allele, pga1g42r, encoding a constitutively active pga1 galpha subunit, was introduced in penicillium roqueforti by transformation, resulting in a phenotype characterized by low sporulation a ...200919784932
evidence for dicer-dependent rna interference in the industrial penicillin producer penicillium chrysogenum.rna interference (rnai) is a sequence-specific post-transcriptional gene silencing system that downregulates target gene expression. here, we provide several lines of evidence for rna silencing in the industrial beta-lactam antibiotic producer penicillium chrysogenum using the dsred reporter gene under the control of the constitutive trpc promoter or the inducible xylp promoter. the functional rnai system was verified by detection of sirnas that hybridized exclusively with gene-specific (32)p-la ...200919797363
jem spotlight: fungi, mycotoxins and microbial volatile organic compounds in mouldy interiors from water-damaged buildings.concerns have been raised about exposure to mycotoxin producing fungi and the microbial volatile organic compounds (mvocs) they produce in indoor environments. therefore, the presence of fungi and mycotoxins was investigated in 99 samples (air, dust, wallpaper, mycelium or silicone) collected in the mouldy interiors of seven water-damaged buildings. in addition, volatile organic compounds (vocs) were sampled. the mycotoxins were analysed by liquid chromatography-tandem mass spectrometry (lc-ms/m ...200919809708
enhanced removal of pentachlorophenol and 2,4-d from aqueous solution by an aminated biosorbent.the fungal biomasses of penicillium chrysogenum were used as raw materials to prepare the aminated adsorbent through chemical grafting of polyethylenimine (pei) on the biomass surface. due to the protonation of amine groups, the pei-modified biomass was found to possess the zero point of zeta potential at ph 10.2, higher than the pristine biomass at ph 3.8. the aminated biosorbent was effective in removing anionic pentachlorophenol (pcp) and 2,4-dichlorophenoxyacetic acid (2,4-d) from aqueous so ...200919013710
moulds and ochratoxin a on surfaces of artisanal and industrial dry sausages.the use of moulds as a seasoning for sausage can have both desirable and undesirable consequences. the desirable consequences are the creation of a successful product that appeals to consumers. the undesirable consequences are due to the growth of undesirable moulds that produce highly toxic secondary metabolites referred to as mycotoxins. the aim of the paper was to investigate the presence of moulds producing ochratoxin a (ota) on the surface of sausages from northern italy. a total of 757 mou ...200919028307
effects on perceived air quality and symptoms of exposure to microbially produced metabolites and compounds emitted from damp building materials.this work investigated perceived air quality and health effects from exposure to low to high levels of volatile organic compounds (vocs) emitted from damp building materials and a mixture of molds growing on the materials. a mixture of wallemia sebi, fusarium culmorum, penicillium chrysogenum, ulocladium botrytis, and aspergillus versicolor was inoculated on pine wood and particle board. in study 1, each of 27 participants took part in two exposure conditions, one with air from molds growing on ...200919077173
purification and characterization of the cold-active alkaline protease from marine cold-adaptive penicillium chrysogenum fs010.an extracellular cold-active alkaline serine protease from penicillium chrysogenum fs010 has been purified. the purification procedure involved: ammonium sulfate precipitation, deae ion-exchange chromatography and sephadex g-100 gel chromatography. sds-page of the purified enzyme indicated a molecular weight of 41,000 +/- 1,000 da. the protease is stable in a ph range of 7.0-9.0 and has a maximum activity at ph 9.0. compared with other industrial proteases, the enzyme shows a high hydrolytic act ...200919116773
isolation and characterisation of polychlorinated biphenyl (pcb) degrading fungi from a historically contaminated soil.abstract:200919138404
aspergillus fumigatus germ tube growth and not conidia ingestion induces expression of inflammatory mediator genes in the human lung epithelial cell line a549.inhalation of conidia is the main cause of invasive pulmonary aspergillosis (ipa) and the respiratory epithelium is the first line of defence. to explore the triggering factor for the inflammatory response to aspergillus fumigatus, the species mainly responsible for ipa, this study analysed the differential expression of three inflammatory genes in a549 cells after challenge with live and killed conidia. the influence of steroids, one of the main risk factors for developing ipa, was also investi ...200919141733
contribution of peroxisomes to penicillin biosynthesis in aspergillus nidulans.peroxisomal localization of the third enzyme of the penicillin biosynthesis pathway of aspergillus nidulans, acyl-coenzyme a:ipn acyltransferase (iat), is mediated by its atypical peroxisomal targeting signal 1 (pts1). however, mislocalization of iat by deletion of either its pts1 or of genes encoding proteins involved in peroxisome formation or transport does not completely abolish penicillin biosynthesis. this is in contrast to the effects of iat mislocalization in penicillium chrysogenum.200919151327
matching the proteome to the genome: the microbody of penicillin-producing penicillium chrysogenum cells.in the filamentous fungus penicillium chrysogenum, microbodies are essential for penicillin biosynthesis. to better understand the role of these organelles in antibiotics production, we determined the matrix enzyme contents of p. chrysogenum microbodies. using a novel in silico approach, we first obtained a catalogue of 200 p. chrysogenum proteins with putative microbody targeting signals (ptss). this included two orthologs of proteins involved in cephalosporin biosynthesis, which we demonstrate ...200919156454
synthesis of sulfated glucosaminides for profiling substrate specificities of sulfatases and fungal beta-n-acetylhexosaminidases.systematic sulfation: sulfated glycoconjugates are degraded either by desulfation followed by glycoside cleavage, or by glycoside cleavage followed by desulfation. to study these processes, here we report the synthesis of four regioisomerically sulfated p-nitrophenyl glucosaminides from the common precursor p-nitrophenyl n-acetyl-beta-d-glucosaminide. these substrates allowed the rapid analysis of the substrate preferences of a set of four sulfatases and 24 hexosaminidases.sulfated carbohydrates ...200919156788
the vacuolar serine protease, a cross-reactive allergen from cladosporium herbarum.subtilisin-like serine proteases make up one of the most important allergen-families regarding the number of individual allergens. previously, fungal subtilisin-like serine proteases have been identified from aspergillus-, penicillium-, and trichophyton-species having a prevalence of ige-reactivity between 33% and 80%. since ige-cross-reactivity is a common phenomenon within fungal species we wanted to know whether this protein also represents an allergen in cladosporium herbarum. hence, a scree ...200919162325
disruption of the nadph-dependent glutamate dehydrogenase affects the morphology of two industrial strains of penicillium chrysogenum.new morphological aspects of penicillium chrysogenum were found during physiological characterisation of two nadph-dependent glutamate dehydrogenase mutant strains. a morphological characterisation of the previously constructed strains, together with the two beta-lactam producing industrial recipient strains, was conducted. the reference strains showed a compact structure with highly branched hyphal elements whereas the morphology of the deltagdha strains consisting of long elongated hyphal elem ...200919167440
exploring and dissecting genome-wide gene expression responses of penicillium chrysogenum to phenylacetic acid consumption and penicilling production.since the discovery of the antibacterial activity of penicillin by fleming 80 years ago, improvements of penicillin titer were essentially achieved by classical strain improvement through mutagenesis and screening. the recent sequencing of penicillium chrysogenum strain wisconsin1255-54 and the availability of genomics tools such as dna-microarray offer new perspective.200919203396
characterization of a phenylacetate-coa ligase from penicillium chrysogenum.enzymatic activation of paa (phenylacetic acid) to phenylacetyl-coa is an important step in the biosynthesis of the beta-lactam antibiotic penicillin g by the fungus penicillium chrysogenum. coa esters of paa and poa (phenoxyacetic acid) act as acyl donors in the exchange of the aminoadipyl side chain of isopenicillin n to produce penicillin g or penicillin v. the phl gene, encoding a pcl (phenylacetate-coa ligase), was cloned in escherichia coli as a maltose-binding protein fusion and the bioch ...200918834333
[analysis of oxygen transfer in bioreactors for fungus broths. 2. suspensions of p. chrysogenum mycelial associations].unlike the p. shermanii and s. cerevisiae cultures, the study on the distribution of oxygen transfer in stirred bioreactor for p. chrysogenum pellets broths indicated that this process is controlled mainly by the deposition tendency of the biomass. similar to the previously studied systems, the analysis of k(1)a distribution indicated its heterogeneity on the bioreactor height, the oxygen transfer rate increasing from position 1 to 4. contrary to the bacterial and yeasts cultures, the intensific ...200920191858
an engineered yeast efficiently secreting penicillin.this study aimed at developing an alternative host for the production of penicillin (pen). as yet, the industrial production of this beta-lactam antibiotic is confined to the filamentous fungus penicillium chrysogenum. as such, the yeast hansenula polymorpha, a recognized producer of pharmaceuticals, represents an attractive alternative. introduction of the p. chrysogenum gene encoding the non-ribosomal peptide synthetase (nrps) delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine synthetase (acvs) ...200920016817
proteolytic activity of beef luncheon fungi as affected by incorporation of some food preservatives.screening of thirty-one fungal isolates (representing 16 genera, 28 species and 3 varieties) collected from beef luncheon meat for their abilities to produce protease enzyme revealed that eleven isolates (35.48%) exhibited high protease production. however, fifteen isolates (48.39%) had moderate ability and 5 isolates (16.13%) were low producers. aspergillus flavus, gibberella fujikuroi and penicillium chrysogenum were the most active producers of protease. the incorporation of five chemical sub ...200920038493
[effects of pa-pex 11 gene on penicillin production in penicillium aurantiogriseum].to study the effect of pa-pex 11 gene on penicillin production in pencillium aurantiogriseum.200920112676
cultivated anti-aspergillus t(h)1 cells.invasive aspergillosis remains a serious complication in patients undergoing allogeneic stem cell transplantation. since it became clear that lymphocytes provide a critical secondary defense against fungi, adoptive transfer of functionally active anti-aspergillus t cells might be an option to restore adaptive immune effector mechanisms. using the interferon (ifn)-gamma secretion assay, we isolated human activated t cells upon stimulation with a cellular extract of aspergillus fumigatus. after a ...200918651313
study on the diversity of endophytic communities from rice (oryza sativa l.) and their antagonistic activities in vitro.endophytic populations were isolated from 2400 segments of oryza sativa collected from bhadra river project area, southern india during december 2005 (winter) and april 2006 (summer). overall colonization rates from surface sterilized tissues were 40.3% in roots and 25.83% in leaves during winter season, 20.15% in roots and 8.66% in leaves during summer season. nineteen different fungal taxa, a streptomyces sp. and bacterial species were isolated. streptomyces sp., chaetomium globosum, penicilli ...200917433644
dynamic gene expression regulation model for growth and penicillin production in penicillium chrysogenum.as is often the case for microbial product formation, the penicillin production rate of penicillium chrysogenum has been observed to be a function of the growth rate of the organism. the relation between the biomass specific rate of penicillin formation (q(p)) and growth rate (mu) has been measured under steady state conditions in carbon limited chemostats resulting in a steady state q(p)(mu) relation. direct application of such a relation to predict the rate of product formation during dynamic ...201020148401
proteome analysis of the penicillin producer penicillium chrysogenum: characterization of protein changes during the industrial strain improvement.proteomics is a powerful tool to understand the molecular mechanisms causing the production of high penicillin titers by industrial strains of the filamentous fungus penicillium chrysogenum as the result of strain improvement programs. penicillin biosynthesis is an excellent model system for many other bioactive microbial metabolites. the recent publication of the p. chrysogenum genome has established the basis to understand the molecular processes underlying penicillin overproduction. we report ...201020154335
deep sequencing analysis of viruses infecting grapevines: virome of a vineyard.double stranded rna, isolated from 44 pooled randomly selected vines from a diseased south african vineyard, has been used in a deep sequencing analysis to build a census of the viral population. the dsrna was sequenced in an unbiased manner using the sequencing-by-synthesis technology offered by the illumina genome analyzer ii and yielded 837 megabases of metagenomic sequence data. four known viral pathogens were identified. it was found that grapevine leafroll-associated virus 3 (glrav-3) is t ...201020172578
use of the weibull model to describe inactivation of dry-harvested conidia of different penicillium species by ethanol vapours.this study aimed at modelling the effect of ethanol vapours, in the range 0.7-7.5 kpa, on the inactivation of dry-harvested conidia of penicillium chrysogenum, penicillium digitatum and penicillium italicum.201020070448
[phylogenetic analysis and detection of selective pressure on the cvnh domains].cyanovirin-n (cv-n) is a novel protein with broad-spectrum antiviral activity. its homologs constitute a protein family known as cvnh (cyanovirin-n homology), which possess the evolutionarily conserved anti-hiv (human immunodeficiency virus) domain. in this study, more details about the patchy organism distribution of cvnh domain were explored by reconstructing gene trees. duplicated cvnh sequences were also identified in a wide range of species including aspergillus niger, neosartorya fischeri ...201020085891
molecular cloning, characterization and expression analysis of two beta-n-acetylhexosaminidase homologs of coccidioides posadasii.two full-length cdnas were isolated from coccidioides posadasii that encode two deduced proteins (cphex1 and cphex2) with homology to the glycosyl hydrolase 20 family of beta-n-acetylhexosaminidases. cphex1 consists of 595 amino acids, has a predicted molecular mass of 68 kda and shares the highest identity with the n-acetylhexosaminidase (naga) of aspergillus nidulans, while cphex2 consists of 603 amino acids, has a predicted molecular mass of 68.5 kda and shares the highest identity with nag1 ...201020109094
the antifungal protein paf interferes with pkc/mpk and camp/pka signalling of aspergillus nidulans.the penicillium chrysogenum antifungal protein paf inhibits polar growth and induces apoptosis in aspergillus nidulans. we report here that two signalling cascades are implicated in its antifungal activity. paf activates the camp/protein kinase a (pka) signalling cascade. a pkaa deletion mutant exhibited reduced sensitivity towards paf. this was substantiated by the use of pharmacological modulators: paf aggravated the effect of the activator 8-br-camp and partially relieved the repressive activ ...201019889092
poly(ethylene glycol) (peg-400) as an alternative reaction solvent for the synthesis of some new 1-(4-(4'-chlorophenyl)-2-thiazolyl)-3-aryl-5-(2-butyl-4-chloro-1h-imidazol-5yl)-2-pyrazolines and their in vitro antimicrobial evaluation.several 1-(4-(4'-chlorophenyl)-2-thiazolyl)-3-aryl-5-(2-butyl-4-chloro-1h-imidazol-5yl)-2-pyrazoline derivatives were prepared by the base catalyzed treatment of appropriate chalcones with 4-(4'-chlorophenyl)-2-hydrazino-thiazole in poly (ethylene glycol) (peg-400) as an alternative reaction solvent. all the synthesized compounds were tested for their antimicrobial activities against escherichia coli (mtcc 2939), salmonella typhi (mtcc 98), staphylococcus aureus (mtcc 96), bacillus subtilis (mtc ...201019896247
characterization of the novel antifungal protein pgafp and the encoding gene of penicillium chrysogenum.the strain rp42c from penicillium chrysogenum produces a small protein pgafp that inhibits the growth of some toxigenic molds. the molecular mass of the protein determined by electrospray ionization mass spectrometry (esi-ms) was 6 494da. pgafp showed a cationic character with an estimated pi value of 9.22. upon chemical and enzymatic treatments of pgafp, no evidence for n- or o-glycosylations was obtained. five partial sequences of pgafp were obtained by edman degradation and by esi-ms/ms after ...201019914321
calculation errors of time-varying flux control coefficients obtained from elasticity coefficients by means of summation and connectivity theorems in metabolic control analysis.this paper investigates the accuracy of a matrix method proposed by other researchers to calculate time-varying flux control coefficients (dynamic fccs) from elasticity coefficients by means of summation and connectivity theorems in the framework of metabolic control analysis. a mathematical model for the fed-batch penicillin v fermentation process is used as a case example for discussion. calculated results reveal that this method produces significant calculation errors because the theorems are ...201019932125
potential for preventing spread of fungi in air-conditioning systems constructed using copper instead of aluminium.as copper has been previously suggested as an antimicrobial surface, we tested the effectiveness of copper as an antifungal surface which could be used in air-conditioning systems as an alternative to aluminium.201019943884
allyl isothiocyanate as a cancer chemopreventive phytochemical.allyl isothiocyanate (aitc), which occurs in many common cruciferous vegetables, is widely and often frequently consumed by humans. besides antimicrobial activity against a wide spectrum of pathogens, it showed anticancer activity in both cultured cancer cells and animal models, although the underlining mechanisms remain largely undefined. bioavailability of aitc is extremely high, as nearly 90% of orally administered aitc is absorbed. aitc absorbed in vivo is metabolized mainly through the merc ...201019960458
cloning, characterization, and engineering of fungal l-arabinitol dehydrogenases.l-arabinitol 4-dehydrogenase (lad) catalyzes the conversion of l-arabinitol to l-xylulose with concomitant nad(+) reduction in fungal l-arabinose catabolism. it is an important enzyme in the development of recombinant organisms that convert l: -arabinose to fuels and chemicals. here, we report the cloning, characterization, and engineering of four fungal lads from penicillium chrysogenum, pichia guilliermondii, aspergillus niger, and trichoderma longibrachiatum, respectively. the lad from p. gui ...201020414651
antimicrobial activity of submerged cultures of chilean basidiomycetes.this study is part of a screening program aimed at searching for bioactive metabolites from chilean basidiomycetes. submerged cultivation of fungal mycelia in liquid media was evaluated for antimicrobial activity. a total of 148 strains were obtained in vitro. the extracts produced from submerged cultures were evaluated against bacteria and fungi. in the primary antimicrobial assay, approximately 60% of the extracts presented positive biological activity. the highest frequencies of active strain ...201020425689
the t=1 capsid protein of penicillium chrysogenum virus is formed by a repeated helix-rich core indicative of gene duplication.penicillium chrysogenum virus (pcv), a member of the chrysoviridae family, is a double-stranded rna (dsrna) fungal virus with a multipartite genome, with each rna molecule encapsidated in a separate particle. chrysoviruses lack an extracellular route and are transmitted during sporogenesis and cell fusion. the pcv capsid, based on a t=1 lattice containing 60 subunits of the 982-amino-acid capsid protein, remains structurally undisturbed throughout the viral cycle, participates in genome metaboli ...201020463071
application of the saccharomyces cerevisiae flp/frt recombination system in filamentous fungi for marker recycling and construction of knockout strains devoid of heterologous genes.to overcome the limited availability of antibiotic resistance markers in filamentous fungi, we adapted the flp/frt recombination system from the yeast saccharomyces cerevisiae for marker recycling. we tested this system in the penicillin producer penicillium chrysogenum using different experimental approaches. in a two-step application, we first integrated ectopically a nourseothricin resistance cassette flanked by the frt sequences in direct repeat orientation (frt-nat1 cassette) into a p. chry ...201020472720
core promoters of the penicillin biosynthesis genes and quantitative rt-pcr analysis of these genes in high and low production strain of penicillium chrysogenum.the transcription start points of the penicillin biosynthesis genes from penicillium chrysogenum were mapped using the primer extension method. for each of the three genes consensus sequences of the core promoter elements were identified, supporting the notion that the basal transcription of these genes is mediated separately. interestingly, transcription start of the pcbc gene is located within the potential inr element with no tata box-like sequence being found at expected position. this is in ...201020490754
intracellular metabolite determination in the presence of extracellular abundance: application to the penicillin biosynthesis pathway in penicillium chrysogenum.important steps in metabolic pathways are formed by the transport of substrates and products over the cell membrane. the study of in vivo transport kinetics requires accurate quantification of intra- and extracellular levels of the transported compounds. especially in case of extracellular abundance, the proper determination of intracellular metabolite levels poses challenges. efficient removal of extracellular substrates and products is therefore important not to overestimate the intracellular ...201020506508
[fungal allergy among art conservators: prevalence, risk factors and clinical symptoms].the aim of the study was to evaluate the prevalence of hypersensitivity to fungi among art conservators and museum workers.201020509551
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