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effects of high-pressure processing on in vitro infectivity of encephalitozoon cuniculi.high-pressure processing (hpp) has been shown to be an effective means of eliminating bacteria and destructive enzymes from a variety of food products. hpp extends the shelf life of products while maintaining the sensory features of food and beverages. in this study, we examined the effects of hpp on the infectivity of encephalitozoon cuniculi spores in vitro. spores were exposed to between 140 and 550 mpa for 1 min in a commercial hpp unit. following treatment, the spores were loaded onto cell ...200516539038
a cytosine-rich region upstream of start codons serving as a signal for initiation of translation in encephalitozoon cuniculi?encephalitozoon cuniculi, an intracellular eukaryote frequently infecting immunodeficient humans, displays pronounced compaction in its genes. short-leaded mrna has been observed which has led to speculation into alternative mechanisms of translation initiation. it has been proposed that a 'downstream box' could serve as an initiation signal. in this report, non-randomness analysis was used to study the genes of e. cuniculi. surprisingly, it was found that the region 5-10nt upstream of start cod ...200515694488
the parasitophorous vacuole membrane of encephalitozoon cuniculi lacks host cell membrane proteins immediately after invasion.microsporidia of the genus encephalitozoon develop inside a parasitophorous vacuole (pv) of unknown origin. using colocalization studies, the pv was found to be absent from the endocytic pathway markers early endosomal autoantigen 1, transferrin receptor, and lysosome-associated membrane protein 1 and for the endoplasmic reticulum marker calnexin. the nonfusiogenic characteristic of the pv appears to be acquired as early as 1 min postinfection and is not reversed by drug treatment with albendazo ...200515643077
a review of the development of two types of human skeletal muscle infections from microsporidia associated with pathology in invertebrates and cold-blooded vertebrates.traditionally, the microsporidia were primarily studied in insects and fish. there were only a few human cases of microsporidiosis reported until the advent of aids, when the number of human microsporidian infections dramatically increased and the importance of these new pathogens to medicine became evident. over a dozen different kinds of microsporidia infecting humans have been reported. while some of these infections were identified in new genera (enterocytozoon, vittaforma), there were also ...200516004364
humoral intestinal immunity against encephalitozoon cuniculi (microsporidia) infection in mice.three strains of mice, balb/c, il-12 knock-out (ko) and inf-gamma knock-out, were chosen as an experimental model for the study of intestinal immunity induction against encephalitozoon cuniculi levaditi, nicolau et schoen, 1923 infection. mice were infected perorally with 10(7) spores and re-infected with the same dose 70 days after the first infection. the anti-e. cuniculi iga, igg and igm responses in sera and extracts of stool samples were determined by elisa. results have shown specific anti ...200516004375
investigations into microsporidian methionine aminopeptidase type 2: a therapeutic target for microsporidiosis.the microsporidia have been reported to cause a wide range of clinical diseases particularly in patients that are immunosuppressed. they can infect virtually any organ system and cases of gastrointestinal infection, encephalitis, ocular infection, sinusitis, myositis and disseminated infection are well described in the literature. while benzimidazoles such as albendazole are active against many species of microsporidia, these drugs do not have significant activity against enterocytozoon bieneusi ...200516004378
the microsporidian polar tube: a highly specialised invasion organelle.all of the members of the microsporidia possess a unique, highly specialised structure, the polar tube. this article reviews the available data on the organisation, structure and function of this invasion organelle. it was over 100 years ago that thelohan accurately described the microsporidian polar tube and the triggering of its discharge. in the spore, the polar tube is connected at the anterior end, and then coils around the sporoplasm. upon appropriate environmental stimulation the polar tu ...200516005007
fatal pulmonary microsporidiosis due to encephalitozoon cuniculi following allogeneic bone marrow transplantation for acute myelogenous leukemia.microsporidia are ubiquitous obligate eukaryotic intracellular parasites that are now felt to be more akin to degenerate fungi than to protozoa. microsporidia can be highly pathogenic, causing a broad range of symptoms in humans, especially individuals who are immunocompromised. the vast majority of human cases of microsporidiosis have been reported during the past 20 years, in patients with hiv/aids, while only relatively rare cases have been described in immunocompetent individuals. however, m ...200516036880
microsporidian polar tube proteins: highly divergent but closely linked genes encode ptp1 and ptp2 in members of the evolutionarily distant antonospora and encephalitozoon groups.the spore polar tube is a unique organelle required for cell invasion by fungi-related microsporidian parasites. two major polar tube proteins (ptp1 and ptp2) are encoded by two tandemly arranged genes in encephalitozoon species. a look at antonospora (nosema) locustae contigs (http://jbpc.mbl.edu/nosema/contigs/) revealed significant conservation in the order and orientation of various genes, despite high sequence divergence features, when comparing with encephalitozoon cuniculi complete genome ...200516051504
simultaneous detection of four human pathogenic microsporidian species from clinical samples by oligonucleotide microarray.microsporidian species have been rapidly emerging as human enteric pathogens in immunocompromised and immunocompetent individuals in recent years. routine diagnostic techniques for microsporidia in clinical laboratories are laborious and insensitive and tend to underestimate their presence. in most instances, they are unable to differentiate species of spores due to their small sizes and similar morphologies. in this study, we report the development of another protozoan oligonucleotide microarra ...200516081959
experimental meningoencephalomyelitis by encephalitozoon cuniculi in cyclophosphamide-immunosuppressed mice.encephalitozoonosis is an increasingly important opportunistic protozoan infection in immunocompromised individuals. this study aims to examine the development of an experimental encephalitozoon cuniculi infection in the central nervous system of immunosuppressed mice. adult balb-c mice were inoculated intraperitoneally with e. cuniculi spores, treated with cyclophosphamide during the experimental period and killed from 15 to 75 days post-inoculation. tissue samples were collected and processed ...200516100970
proteolytic activity in encephalitozoon cuniculi sporogonial stages: predominance of metallopeptidases including an aminopeptidase-p-like enzyme.a fraction enriched in spore precursor cells (sporoblasts) of the microsporidian encephalitozoon cuniculi, an intracellular parasite of mammals, was obtained by percoll gradient centrifugation. soluble extracts of these cells exhibited proteolytic activity towards azocasein, with an alkaline optimum ph range (9-10). prevalence of some metallopeptidases was supported by the stimulating effect of ca2+, mg2+, mn2+ and zn2+ ions, and inhibition by two chelating agents (edta and 1,10-phenanthroline), ...200516137693
effects of a novel anti-exospore monoclonal antibody on microsporidial development in vitro.the possible role of humoral antibodies in the immune response to microsporidiosis was studied using a novel anti-exospore monoclonal antibody (mab) p5/h1 which recognised encephalitozoon cuniculi. the effect of the p5/h1 mab on microsporidial growth in vitro resulted in a reduction of the numbers of e. cuniculi spores in a vero e6 cell-line culture. this reduction in the number of infected cells and the decrease of intracellular spores in infected cells was found when mab p5/h1 was present in c ...200414610668
what does the microsporidian e. cuniculi tell us about the origin of the eukaryotic cell?the relationship among the three cellular domains archaea, bacteria, and eukarya has become a central problem in unraveling the tree of life. this relationship can now be studied as the completely sequenced genomes of representatives of these cellular domains become available. we performed a bioinformatic investigation of the encephalitozoon cuniculi proteome. e. cuniculi has the smallest sequenced eukaryotic genome, 2.9 megabases coding for 1997 proteins. the proteins of e. cuniculi were compar ...200415693625
susceptibility of ifn-gamma or il-12 knock-out and scid mice to infection with two microsporidian species, encephalitozoon cuniculi and e. intestinalis.susceptibility of three strains of immunodeficient mice to two related microsporidian species encephalitozoon cuniculi levaditi, nicolau et schoen, 1923 and encephalitozoon intestinalis (cali, kotler et orenstein, 1993) was compared. while both, severe combined immunodeficient (scid) and interferon-gamma knock-out (ifn-gamma ko) mice, succumbed to either intraperitoneal (i.p.) or peroral (p.o.) (natural) infection with both parasites, only i.p. infection with e. cuniculi killed interleukin-12 kn ...200415729937
[host immune response in mammals to encephalitozoon cuniculi infection].the immunity system status of an individual plays the key role in regulation of opportune infection. in the fight against the intracellular parasites several non-specific as well as specific immunity mechanisms are applied. the dominant role in response to infection caused by the representatives of genus encephalitozoon plays the cell-mediated immune response. encephalitozoon cuniculi, as the most explored representative of this genus is able to survive in the host organism despite his active im ...200415524273
[laboratory diagnostics of human microsporidiosis].there are many techniques available for the identification of microsporidia in clinical specimens. chromotrope 2r, calcofluor white m2r and fish technique have all been reported to be useful as selective methods for microsporidia in stool specimens and in body fluids. microsporidia in histologic tissue preparations have also been visualized with giemsa, hematoxylin and eosin stain, brown-hopps stain or warthin-starry staining. microsporidia can also be identified by using tests for detecting igg ...200416862802
polyamine metabolism in a member of the phylum microspora (encephalitozoon cuniculi): effects of polyamine analogues.the uptake, biosynthesis and catabolism of polyamines in the microsporidian parasite encephalitozoon cuniculi are detailed with reference to the effects of oligoamine and arylamine analogues of polyamines. enc. cuniculi, an intracellular parasite of mammalian cells, has both biosynthetic and catabolic enzymes of polyamine metabolism, as demonstrated in cell-free extracts of mature spores. the uptake of polyamines was measured in immature, pre-emergent spores isolated from host cells by percoll g ...200415133083
structure and mechanism of mrna cap (guanine-n7) methyltransferase.a suite of crystal structures is reported for a cellular mrna cap (guanine-n7) methyltransferase in complex with adomet, adohcy, and the cap guanylate. superposition of ligand complexes suggests an in-line mechanism of methyl transfer, albeit without direct contacts between the enzyme and either the n7 atom of guanine (the attacking nucleophile), the methyl carbon of adomet, or the sulfur of adomet/adohcy (the leaving group). the structures indicate that catalysis of cap n7 methylation is accomp ...200414731396
a comprehensive evolutionary classification of proteins encoded in complete eukaryotic genomes.sequencing the genomes of multiple, taxonomically diverse eukaryotes enables in-depth comparative-genomic analysis which is expected to help in reconstructing ancestral eukaryotic genomes and major events in eukaryotic evolution and in making functional predictions for currently uncharacterized conserved genes.200414759257
guanosine distribution and oxidation resistance in eight eukaryotic genomes.reactive oxygen species that attack dna are continuously generated in living cells. both the guanosine (g) mole fraction and its distribution should affect the stability of genomes and their parts to oxidation. at a lesser g content, genomes should be more oxidation resistant or "ennobled". oxidant scavenging by g's in nonessential parts of introns and intergenic domains should decrease g oxidation in the essential exons. to determine whether genomes are indeed ennobled and whether oxidant-scave ...200414982441
disseminated lethal encephalitozoon cuniculi (genotype iii) infections in cotton-top tamarins (oedipomidas oedipus)--a case report.for the first time, encephalitozoon (e.) cuniculi genotype iii ('dog strain') was verified in two cotton-top tamarins (oedipomidas oedipus) by light microscopy, immunohistochemistry, electron microscopy, pcr and sequencing. the animals had a disseminated lethal infection with this protist. in earlier reports, genotype iii had been found only in domestic dogs, man, emperor tamarins (saguinus imperator) and golden lion tamarins (leontopithecus rosalia). this investigation establishes now that the ...200414984833
immune response in mice infected by encephalitozoon cuniculi and suppressed by dexamethasone.several indicators of immune response were observed in immunocompetent mice of the icr line and those suppressed by dexamethasone upon their experimental infection with the microsporidia of encephalitozoon cuniculi. the mice were infected by one-shot intraperitoneal administration of 5 x 10(7) pathogenic spores. on days 7, 14, 28 and 42 after infection, peripheral blood leukocyte phagocytic activity was determined and compared, including phagocytic index and the blastogenic response in spleen ce ...200415119788
phylogenetic analysis of the complete genome sequence of encephalitozoon cuniculi supports the fungal origin of microsporidia and reveals a high frequency of fast-evolving genes.microsporidia are unicellular eukaryotes living as obligate intracellular parasites. lacking mitochondria, they were initially considered as having diverged before the endosymbiosis at the origin of mitochondria. that microsporidia were primitively amitochondriate was first questioned by the discovery of microsporidial sequences homologous to genes encoding mitochondrial proteins and then refuted by the identification of remnants of mitochondria in their cytoplasm. various molecular phylogenies ...200415599510
prevalence of antibodies to encephalitozoon cuniculi in horses in the israel.infection with the intracellular microsporidium encephalitozoon cuniculi can cause a serious disease--encephalitozoonosis in various animals and people. several species of mammals, including the horse, were seem to be potential sources of encephalitozoonosis for animal as well as human hosts. the disease diagnosis is based on clinical signs, pathological findings, and the detection of e. cuniculi or circulating antibodies directed against the parasite. this study investigates the seroconversion ...200415627335
serological survey of selected pathogens of free-ranging foxes in southern argentina, 1998--2001.a serological survey was conducted in the patagonia region of argentina to estimate the prevalence of nine disease agents within the populations of free-ranging culpeo (dusicyon culpaeus) and grey (dusicyon griseus) foxes. the disease agents were aujeszky's disease virus (adv), brucella, canine adenovirus (cav), canine distemper virus (cdv), canine parvovirus (cpv), encephalitozoon cuniculi, leptospira, neospora caninum, and toxoplasma gondii. a total of 84 foxes were sampled (28 culpeo and 56 g ...200415861875
an encephalitozoon cuniculi ortholog of the rna polymerase ii carboxyl-terminal domain (ctd) serine phosphatase fcp1.fcp1 is an essential protein serine phosphatase that dephosphorylates ser2 or ser5 of the rna polymerase ii carboxyl-terminal domain (ctd) heptad repeat y(1)s(2)p(3)t(4)s(5)p(6)s(7). the ctd of the microsporidian parasite encephalitozoon cuniculi consists of 15 heptad repeats, which approximates the minimal ctd length requirement for cell viability in yeast. here we show that e. cuniculi encodes a minimized 411-aa fcp1-like protein (ecfcp1), which consists of a dxdx(t/v) phosphatase domain and a ...200415170348
serodiagnostic studies in an immunocompetent individual infected with encephalitozoon cuniculi.little is known about the prevalence and clinical significance of infection with encephalitozoon species in immunocompetent individuals. in the present study, by using indirect immunofluorescence technique (ifat), western blot, and recombinant antigens of the spore wall (swp1) and polar tube (ptp1, ptp2, and ptp3 ), we analyzed the igg antibody response of a laboratory worker who was infected with encephalitozoon cuniculi. serum samples were analyzed 1, 20, 32, and 38 months after infection. aft ...200415181572
genome compaction and stability in microsporidian intracellular parasites.microsporidian genomes are extraordinary among eukaryotes for their extreme reduction: although they are similar in form to other eukaryotic genomes, they are typically smaller than many prokaryotic genomes. at the same time, their rates of sequence evolution are among the highest for eukaryotic organisms. to explore the effects of compaction on nuclear genome evolution, we sequenced 685,000 bp of the antonospora locustae genome (formerly nosema locustae) and compared its organization with the r ...200415186746
in vitro effects of resveratrol on the viability and infectivity of the microsporidian encephalitozoon cuniculi.microsporidians of the genus encephalitozoon are an important cause of disease in immunocompromised patients, and there are currently no completely effective treatments. the present study investigated the viability and infectivity of spores of encephalitozoon cuniculi that had been exposed to resveratrol (resv), a natural phytoalexin found in grapes and red wine. resv at 50 microm showed significant sporicidal activity, and at 10 to 50 microm it reduced the capacity of the spores to infect dog k ...200415215100
a comparison of homologous genes encoding aminopeptidases among bird and human encephalitozoon hellem isolates and a rabbit e. cuniculi isolate.encephalitozoon cuniculi and e. hellem are often recognized as the agents of human microsporidiosis, but less than optimal therapy is available for treatment. the identification of enzymes critical to the parasitic life cycle is an important step in finding targets for potential drug development. aminopeptidase gene sequences were obtained from cdna and gdna from avian and human e. hellem isolates and from a rabbit e. cuniculi isolate. at the amino acid level, the aminopeptidase sequences from t ...200415221463
quantitation of microsporidia in cultured cells by flow cytometry.microsporidia are obligate intracellular protozoan parasites that emerged as major opportunistic pathogens in humans since the onset of the aids pandemic. in the present study, we investigated whether fcm is a useful method for the quantitation of intracellular microsporidian spores in cultured cells.200415229863
class ii photolyase in a microsporidian intracellular parasite.photoreactivation is the repair of dna damage induced by ultraviolet light radiation using the energy contained in visible-light photons. the process is carried out by a single enzyme, photolyase, which is part of a large and ancient photolyase/cryptochrome gene family. we have characterised a photolyase gene from the microsporidian parasite, antonospora locustae (formerly nosema locustae) and show that it encodes a functional photoreactivating enzyme and is expressed in the infectious spore sta ...200415288781
comparative analysis of complete genomes reveals gene loss, acquisition and acceleration of evolutionary rates in metazoa, suggests a prevalence of evolution via gene acquisition and indicates that the evolutionary rates in animals tend to be conserved.in this study we systematically examined the differences between the proteomes of metazoa and other eukaryotes. metazoans (homo sapiens, ceanorhabditis elegans and drosophila melanogaster) were compared with a plant (arabidopsis thaliana), fungi (saccharomyces cerevisiae and schizosaccaromyces pombe) and encephalitozoan cuniculi. we identified 159 gene families that were probably lost in the metazoan branch and 1263 orthologous families that were specific to metazoa and were likely to have origi ...200415448184
the genes encoding camp-dependent protein kinase catalytic subunit homologues of the microsporidia encephalitozoon intestinalis and e. cuniculi: molecular characterisation and phylogenetic analysis.a gene encoding a protein kinase was identified by homology-based pcr amplification in encephalitozoon intestinalis, a microsporidian parasite pathogenic to humans, and its orthologue has been identified by database mining in the genome of the related species e. cuniculi, whose sequence has been recently published. phylogenetic analysis revealed that the proteins encoded by these genes are homologues of the camp-dependent protein kinase catalytic subunits (pkac). southern blot analysis indicated ...200415464436
can heterologous gene expression shed (a torch) light on protein function? 200415491798
monoclonal antibodies for specific detection of encephalitozoon cuniculi.seven species-specific monoclonal antibodies (mabs) were produced against encephalitozoon cuniculi and characterized. the mabs were immunoglobulin g, and when used for indirect microimmunofluorescence microscopy and western immunoblot assays, they detected e. cuniculi originating from clinical samples. they did not cross-react with other encephalitozoon species (e. intestinalis and e. hellem) or with a collection of gram-negative bacteria, yeast, and other parasites. the mabs reacted primarily w ...200415539506
induction of a rapid and strong antigen-specific intraepithelial lymphocyte response during oral encephalitozoon cuniculi infection.encephalitozoon cuniculi continues to pose a problem for immunocompromised patients. previous studies from our laboratory have elucidated the importance of the cd8(+) t cell subset in the protection against systemic parasite infection. there have been no studies related to the mucosal immunity induced against this orally acquired pathogen. in the present study, the immune response generated in the gut after oral e. cuniculi infection was evaluated. an early and rapid increase of the intraepithel ...200415034055
comparative genomics of pneumocystis carinii with other protists: implications for life style.three protistan genomes were analyzed for differential genetic traits that may be associated with biological adaptations to their unique life styles. the microsporidian, encephalitozoon cuniculi, an obligate intracellular parasite; the ascomycetes, pneumocystis carinii, considered an opportunistic pathogen; and saccharomyces cerevisiae, a model organism exhibiting a free-living life style, were used in comparisons of genomic architecture, reproductive strategies, and metabolic capacity predicted ...200415068263
infectivity of microsporidia spores stored in seawater at environmental temperatures.to determine how long spores of encephalitozoon cuniculi, e. hellem, and e. intestinalis remain viable in seawater at environmental temperatures, culture-derived spores were stored in 10, 20, and 30 ppt artificial seawater at 10 and 20 c. at intervals of 1, 2, 4, 8, and 12 wk, spores were tested for infectivity in monolayer cultures of madin darby bovine kidney cells. spores of e. hellem appeared the most robust, some remaining infectious in 30 ppt seawater at 10 c for 12 wk and in 30 ppt seawat ...200415270118
investigations into shaking mink syndrome: an encephalomyelitis of unknown cause in farmed mink (mustela vison) kits in scandinavia.an apparently novel neurological disease clinically characterized by shaking, tremors, seizures, staggering gait, and ataxia was first observed in farmed mink kits in denmark in 2000 and subsequently in sweden, denmark, and finland in 2001, and again in denmark in 2002. lymphoplasmacytic encephalomyelitis was found in the affected kits. the lesions were most severe in the brainstem and cerebellum and consisted of neuronal degeneration and necrosis, neuronophagia, focal and diffuse gliosis, periv ...200415305741
survey for encephalitozoon cuniculi in arctic foxes (alopex lagopus) in greenland.wild arctic foxes (alopex lagopus) from greenland were tested for antibodies to encephalitozoon cuniculi with an enzyme-linked immunosorbent assay and a carbon immunoassay. of 230 tested foxes none was seropositive. this finding contrasts with observations from other arctic areas and absence of rodents in the diet of these arctic foxes is the most likely explanation for absence of e. cuniculi.200312685089
pathologic quiz case: a 45-year-old renal transplant recipient with persistent fever. 200312683908
serological investigation of canine encephalitozoonosis in norway.encephalitozoon cuniculi, a microsporidian parasite of vertebrates, is considered a health risk to aids patients and other immunocompromised human beings. in most hosts, infection with the parasite runs a subclinical course. in some carnivore species, however, clinical disease affecting whole litters arises from intrauterine transmission of the parasite. in both blue foxes ( alopex lagpus) and dogs ( canis familiaris), outbreaks of encephalitozoonosis can be severe. canine encephalitooonosis has ...200312474043
comparison of uv inactivation of spores of three encephalitozoon species with that of spores of two dna repair-deficient bacillus subtilis biodosimetry strains.when exposed to 254-nm uv, spores of encephalitozoon intestinalis, encephalitozoon cuniculi, and encephalitozoon hellem exhibited 3.2-log reductions in viability at uv fluences of 60, 140, and 190 j/m(2), respectively, and demonstrated uv inactivation kinetics similar to those observed for endospores of dna repair-defective mutant bacillus subtilis strains used as biodosimetry surrogates. the results indicate that spores of encephalitozoon spp. are readily inactivated at low uv fluences and that ...200312514061
chlorine inactivation of spores of encephalitozoon spp.this report is an extension of a preliminary investigation on the use of chlorine to inactivate spores of encephalitozoon intestinalis and to investigate the effect of chlorine on two other species, e cuniculi and e. hellem, associated with human infection. the 50% tissue culture infective doses of these three species were also determined. on the basis of the results obtained, it appears that chlorination of water is an effective means of controlling spores of these organisms in the aquatic envi ...200312571067
the homeobox genes of encephalitozoon cuniculi (microsporidia) reveal a putative mating-type locus.homeobox genes have been found in animals, fungi, and plants. recently, the complete genomic sequence of encephalitozoon cuniculi has become available and it was shown that this microsporida species is related to fungi. given this close relatedness to fungi, i have searched the genome of e. cuniculifor homeobox genes. there are 12 homeobox genes as well as one ste12 orthologue. the large number of homeobox genes when compared to the annotations, which only list one, suggests that possibly other ...200312590353
a comprehensive view on proteasomal sequences: implications for the evolution of the proteasome.proteasomes are large multimeric self-compartmentizing proteases, which play a crucial role in the clearance of misfolded proteins, breakdown of regulatory proteins, processing of proteins by specific partial proteolysis, cell cycle control as well as preparation of peptides for immune presentation. two main types can be distinguished by their different tertiary structure: the 20s proteasome and the proteasome-like heat shock protein encoded by heat shock locus v, hslv. usually, each biological ...200312595256
infectivity of microsporidia spores stored in water at environmental temperatures.to determine how long waterborne spores of encephalitozoon cuniculi, e. hellem, and e. intestinalis could survive at environmental temperatures, culture-derived spores were stored in water at 10, 15, 20, 25, and 30 c and tested for infectivity in monolayer cultures of madin darby bovine kidney (mdbk) cells. at 10 c, spores of e. intestinalis were still infective after 12 mo, whereas those of e. hellem and e. cuniculi were infective for 9 and 3 mo, respectively. at 15 c, spores of the same specie ...200312659327
using ancestral sequences to uncover potential gene homologues.gene homologues between distantly related species can be difficult to identify. we test the idea that inferred ancestral sequences could aid in finding gene homologues. ancestral sequences are inferred by aligning gene homologues on a known tree and estimating the most likely amino acid for each position at each node in that tree. blast(r) and hmmer are used separately and together with ancestral sequences to search the genome sequence databases of encephalitozoon cuniculi, entamoeba histolytica ...200315130821
the cog database: an updated version includes eukaryotes.the availability of multiple, essentially complete genome sequences of prokaryotes and eukaryotes spurred both the demand and the opportunity for the construction of an evolutionary classification of genes from these genomes. such a classification system based on orthologous relationships between genes appears to be a natural framework for comparative genomics and should facilitate both functional annotation of genomes and large-scale evolutionary studies.200312969510
encephalitozoon cuniculi in pet rabbits.the results of a serological test for encephalitozoon cuniculi in 125 pet rabbits are reviewed, together with follow-up studies of clinical cases. blood samples were taken from 38 asymptomatic rabbits and 87 rabbits showing neurological, renal or ocular signs suggestive of encephalitozoonosis. in the asymptomatic group, six of 26 (23 per cent) apparently healthy rabbits, sampled as part of a health screen, were seropositive; of the remaining 12 asymptomatic rabbits, sampled because they lived wi ...200312708591
humoral immune response in adult blue foxes (alopex lagopus) after oral infection with encephalitozoon cuniculi spores.encephalitozoon cuniculi causes severe diseases in blue fox puppies. when pregnant vixens are infected, parasites are transmitted over the placenta to the unborn that subsequently develop encephalitozoonosis. adult foxes themselves do not have signs of disease, but show antibody titres to e. cuniculi. the purpose of the present study was to gain information on the immune response in adult foxes after experimental infection. sixteen foxes were infected orally with e. cuniculi spores, eight of the ...200312719134
disseminated encephalitozoon cuniculi infection in a mexican kidney transplant recipient.no cases of encephalitozoon cuniculi infection have been reported in transplant patients.200312811252
overt fatal and chronic subclinical encephalitozoon cuniculi microsporidiosis in a colony of captive emperor tamarins (saguinus imperator).the course of an infection with the microsporidian encephalitozoon cuniculi in a colony of captive emperor tamarins (saguinus imperator) is described. in two litters, the infection was associated with overt disease and death of all infants. immunohistochemistry for e. cuniculi showed generalized infections, and histopathologic evaluation revealed systemic vasculitis and disseminated mixed inflammatory cell infiltration with and without necrosis in several organs. serologically, some of the juven ...200312823634
patterns of context-dependent codon biases.the association of codon context and codon usage was studied in seven bacteria as well as schizosaccharomyces pombe and encephalitozoon cuniculi. the association is strongest in magnitude closest to the codons of interest but there is apparently no rule about which of the two contexts is generally strongest associated to codon usage. in all bacterial species and in the intron-rich sch. pombe it was furthermore observed from plots of chi2 versus n that the wobble positions of codons in the proxim ...200312705888
strand compositional asymmetries of nuclear dna in eukaryotes.both dna replication and transcription are structurally asymmetric processes. an asymmetric nucleotide substitution pattern has been observed between the leading and the lagging strand, and between the coding and the noncoding strand, in eubacterial, viral, and organelle genomes. similar studies in eukaryotes have been rare, because the origins of replication in nuclear genomes are mostly unknown and the replicons are much shorter than those of prokaryotes. to circumvent these predicaments, all ...200314629042
gene loss, protein sequence divergence, gene dispensability, expression level, and interactivity are correlated in eukaryotic evolution.lineage-specific gene loss, to a large extent, accounts for the differences in gene repertoires between genomes, particularly among eukaryotes. we derived a parsimonious scenario of gene losses for eukaryotic orthologous groups (kogs) from seven complete eukaryotic genomes. the scenario involves substantial gene loss in fungi, nematodes, and insects. based on this evolutionary scenario and estimates of the divergence times between major eukaryotic phyla, we introduce a numerical measure, the pro ...200314525925
in utero transmission of encephalitozoon cuniculi strain type i in rabbits.pregnant rabbits were serologically diagnosed as having been infected with encephalitozoon cuniculi. at necropsy at 28 days of gestation, does, placentas and fetuses were found to be infected with e. cuniculi strain type i as evidenced by using the nested-polymerase chain reaction (pcr) technique, thereby confirming vertical transplacental transmission.200312689424
[microsporidial infections in immunocompromised hospitalized patients].microsporidia encephalitozoon cuniculi, obligate intracellular sporeforming protozoan parasite infecting a wide range of vertebrate and invertebrate host, has been described as opportunistic pathogen in severely immunocompromised human host. an antibody immune response to the presence of antibodies against encephalitozoon cuniculi was investigated serologically using indirect immunofluorescence antibody test (ifat). of the total number of 113 sera from immunologically compromised patients examin ...200312822539
analysis of four human microsporidian isolates by maldi-tof mass spectrometry.spores of four species of microsporidia isolated from humans were analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry (maldi-tof ms) and specific biomarkers were found for each. the microsporidia analyzed included three species, encephalitozoon cuniculi, encephalitozoon hellem, and encephalitozoon intestinalis and the fourth organism is the recently described brachiola algerae. whole spores, spore shells, and soluble fractions were applied directly to the mal ...200312836871
serological screening of occurrence of antibodies to encephalitozoon cuniculi in humans and animals in eastern slovakia.encephalitozoon cuniculi is one of the mamalian microsporidian pathogens that can affect a number of different species of animals as well as humans. the presence of specific serum antibodies to encephalitozoon cuniculi was studied in a group of animals and humans from eastern slovakia by the indirect immunofluorescence of antibodies (ifa). 456 people, 571 rabbits, 457 mice, 193 dogs, 72 cats, and 59 sheep were examined. specific anti-e. cuniculi antibodies were found in 26 out of 456 human sera ...200312852743
bacterial catalase in the microsporidian nosema locustae: implications for microsporidian metabolism and genome evolution.microsporidia constitute a group of extremely specialized intracellular parasites that infect virtually all animals. they are highly derived, reduced fungi that lack several features typical of other eukaryotes, including canonical mitochondria, flagella, and peroxisomes. consistent with the absence of peroxisomes in microsporidia, the recently completed genome of the microsporidian encephalitozoon cuniculi lacks a gene for catalase, the major enzymatic marker for the organelle. we show, however ...200314555490
immune response of e. cuniculi infected mice to aflatoxin b1.the immunomodulatory effects of aflatoxin b1 (afb1) on mice experimentally infected with encephalitozoon cuniculi (e. cuniculi) were studied. mice inoculated intraperitoneally with spores of e. cuniculi drank a daily solution of afb1 (0.2 mg kg(-1) of body weight) for 27 days. application of afb1 to mice demonstrated a decrease of immunocompetent cells. on the other hand, the mice infected with e. cuniculi and given afb1 showed significant increased number of both monocytes and cd8+ t cells, and ...200319180805
encephalitozoon cuniculi placentitis and abortion in a quarterhorse mare.encephalitozoon cuniculi is a microsporidial parasite, which has rarely been reported to cause placentitis in animals. a late-term aborted fetus and placenta from a quarterhorse were presented to the livestock disease diagnostic center, university of kentucky, for diagnostic examination. there was a necrotizing placentitis, with distension of many chorionic epithelial cells by intracytoplasmic vacuoles containing 1-2-microm-diameter, elongated, gram-positive organisms. the organisms were identif ...200312580298
novel synthetic polyamines are effective in the treatment of experimental microsporidiosis, an opportunistic aids-associated infection.microsporidia are eukaryotic obligate intracellular protists that are emerging pathogens in immunocompromised hosts, such as patients with aids or patients who have undergone organ transplantation. we have demonstrated in vitro and in vivo that synthetic polyamine analogs are effective antimicrosporidial agents with a broad therapeutic window. cd8-knockout mice or nude mice infected with the microsporidian encephalitozoon cuniculi were cured when they were treated with four different novel polya ...200211751111
fox encephalitozoonosis: isolation of the agent from an outbreak in farmed blue foxes (alopex lagopus) in finland and some hitherto unreported pathologic lesions.the farmed blue fox (alopex lagopus) is particularly susceptible to congenital infections of the microsporidian species encephalitozoon cuniculi. this report is based on an outbreak of the disease in finland with high mortality. five pups (four males and one female) with prolonged disease were examined. the pups had moderate pathological alterations in the kidneys and mild lesions were found in the brains, hearts, salivary and prostatic glands. diagnosis of e. cuniculi infection was made from se ...200212449250
[experimental encephalitozoon cuniculi infection in dexamethasone-immunosuppressed mice].microsporidian encephalitozoon cuniculi has been recognized as an opportunistic pathogen in immunosuppressed individuals, such as aids patients. the objective of the study was to develop pharmacologically immunosuppressed animals as a model of the natural occurring e. cuniculi infection.200212471388
characterization of the mrna capping apparatus of the microsporidian parasite encephalitozoon cuniculi.a scheme of eukaryotic phylogeny has been suggested based on the structure and physical linkage of the enzymes that catalyze mrna cap formation. here we show that the intracellular parasite encephalitozoon cuniculi encodes a complete mrna capping apparatus consisting of separate triphosphatase (eccet1), guanylyltransferase (ecceg1), and methyltransferase (ecm1) enzymes, which we characterize biochemically and genetically. the triphosphatase eccet1 belongs to a metal-dependent phosphohydrolase fa ...200211687593
encephalitozoon cuniculi in rabbits. 200212498418
first isolation and characterisation of encephalitozoon cuniculi from a free-ranging rat (rattus norvegicus).the microsporidian species encephalitozoon cuniculi can infect a wide variety of mammals including man. it is a common parasite in rabbits and several sporadic infections in laboratory rats have been described. based on molecular data three e. cuniculi strains have been identified. here we describe the first in vitro propagation of e. cuniculi, which was isolated from a free-ranging rat (rattus norvegicus). the rat was one of three seropositive animals among 23 rats captured in the city of zuric ...200212163239
effects of gamma radiation on viability of encephalitozoon spores.spores of encephalitozoon cuniculi, e. hellem, and e. intestinalis harvested from cultured mammalian cells were suspended in deionized water, exposed to gamma irradiation at doses of 0-3.0 kgy, and then tested for infectivity by inoculating spores into monolayer cultures of madin-darby bovine kidney cells. the cultures were examined for developing microsporidia 4 days later. as the dosage level of radiation increased, corresponding decreases were observed in the number of developing microsporidi ...200212197142
disseminated microsporidiosis in a renal transplant recipient.disseminated microsporidiosis is diagnosed uncommonly in patients not infected with human immunodeficiency virus (hiv). we present a case of disseminated microsporidiosis in a renal transplant recipient who was seronegative for hiv. chromotrope-based stains were positive for microsporidia in urine, stools, sputum, and conjunctival scrapings. electron microscopy, immunofluorescence, polymerase chain reaction, and cultures of renal tissue identified the organism as encephalitozoon cuniculi. the pa ...200212220248
in vitro activity of polyoxin d and nikkomycin z against encephalitozoon cuniculi.microsporidia of the genus encephalitozoon are emerging protozoal agents that mainly infect immunocompromised patients with aids. at present, disseminated infections with members of the genus encephalitozoon can only be successfully treated with albendazole. as chitin is a basic component of the microsporidian spore. we evaluated, in vitro, the susceptibility of a human-derived strain of encephalitozoon cuniculi to polyoxin d and nikkomycin z, which are known competitive inhibitors of chitin syn ...200212049463
the microsporidian polar tube: evidence for a third polar tube protein (ptp3) in encephalitozoon cuniculi.the invasion strategy used by microsporidia is primarily related to spore germination. small differentiated spores of these fungi-related parasites inject their contents into target cells through the lumen of a rapidly extruded polar tube, as a prerequisite to obligate intracellular development. previous studies in encephalitozoon species that infect mammals have identified two major antigenic polar tube proteins (ptp1 and ptp2) which are predicted to contribute to the high tensile strength of t ...200212076771
an indirect elisa for detection of encephalitozoon cuniculi infection in farmed blue foxes (alopex lagopus).infection with the intracellular microsporidium encephalitozoon cuniculi can cause serious disease, encephalitozoonosis, in the blue fox (alopex lagopus). the disease diagnosis is based on clinical signs and pathological findings, and detection of e. cuniculi or circulating antibodies directed against the parasite. indirect immunofluorescence (ifat) and carbon immunoassay (cia) are the most commonly used serological methods for diagnosis in this species. in the present study, an indirect elisa ( ...200212831174
[encephalitozoon cuniculi and encephalitozoon intestinalis--causes of opportunistic infections].microsporidia (phylum microsporidia) are intracellular parasites that infect a wide range of protozoa, invertebrates and vertebrate hosts. over a 1000 species have been classified into approximately 100 genera. historically, microsporidial infections in silkworms, honey bees, and salmonid fish have been responsible for significant economic losses. more recently, microsporidiosis has been recognized as an important opportunistic infection in immunologically compromised patients. in this review th ...200211881297
comparative analysis of sequences encoding abc systems in the genome of the microsporidian encephalitozoon cuniculi.microsporidia are amitochondriate eukaryotic microbes with fungal affinities and a common status of obligate intracellular parasites. a set of 13 potential genes encoding atp-binding cassette (abc) systems was identified in the fully sequenced genome of encephalitozoon cuniculi. our analyses of multiple alignments, phylogenetic trees and conserved motifs support a distribution of e. cuniculi abc systems within only four subfamilies. six half transporters are homologous to the yeast atm1 mitochon ...200212023075
in vitro cultivation of microsporidia of clinical importance.although attempts to develop methods for the in vitro cultivation of microsporidia began as early as 1937, the interest in the culture of these organisms was confined mostly to microsporidia that infect insects. the successful cultivation in 1969 of encephalitozoon cuniculi, a microsporidium of mammalian origin, and the subsequent identification of these organisms as agents of human disease heightened interest in the cultivation of microsporidia. i describe the methodology as well as the cell li ...200212097248
antigenic diversity of encephalitozoon hellem demonstrated by subspecies-specific monoclonal antibodies.encephalitozoon hellem is a unicellular, obligate intracellular microsporidian species detected and isolated in hiv-infected patients presenting with keratoconjunctivitis, sinusitis, tracheobronchitis, nephritis, cystitis, and disseminated infection. a total of 24 monoclonal antibodies were produced against e. hellem and characterized. the monoclonal antibodies were of the immunoglobulin (ig) g and ig m subclasses, and, when incorporated into indirect immunofluorescence and immunoblotting assays ...200212120990
[serologic test in rabbits with encephalitozoon cuniculi infection]. 200212125166
comparison of the significance of cd4+ and cd8+ t lymphocytes in the protection of mice against encephalitozoon cuniculi infection.the role of t lymphocyte subpopulations in the protection against intraperitoneal (i.p.) and peroral encephalitozoon cuniculi infections was compared in adoptive-transfer experiments using severe combined immunodeficient mice. whereas cd8+ t cell-depleted, but not cd4+ t cell-depleted, balb/c splenocytes failed to protect the mice against i.p. infection, only scid mice reconstituted with both cd4+ t lymphocyte- and cd8+ t lymphocyte-depleted splenocytes succumbed to peroral infection. the result ...200212197135
phacoemulsification for the management of encephalitozoon cuniculi-induced phacoclastic uveitis in a rabbit.phacoemulsification was performed on a new zealand white rabbit with slowly progressive unilateral phacoclastic uveitis and cataract formation. the irrigating solution with lenticular contents were centrifuged and examined cytologically using weber's chromotrope-based stain. microsporidial spores were observed and positively identified as encephalitozoon cuniculi via polymerase chain reaction. more than 1 year following surgical therapy, the rabbit is visual and comfortable without medications.200212236874
functional and evolutionary analysis of a eukaryotic parasitic genome.the dna sequences of the 11 linear chromosomes of the approximately 2.9 mbp genome of encephalitozoon cuniculi, an obligate intracellular parasite of mammals, include approximately 2000 putative protein-coding genes. the compactness of this genome is associated with the length reduction of various genes. essential functions are dependent on a minimal set of genes. phylogenetic analysis supports the hypotheses that microsporidia are related to fungi and have retained a mitochondrion-derived organ ...200212354558
genotyping of encephalitozoon cuniculi isolates found in hokkaido. 200212403911
disseminated microsporidiosis caused by encephalitozoon cuniculi iii (dog type) in an italian aids patient: a retrospective study.we report a case of disseminated microsporidiosis in an italian woman with aids. this study was done retrospectively using formalin-fixed, paraffin-embedded tissue specimens obtained at autopsy. microsporidia spores were found in the necrotic lesions of the liver, kidney, and adrenal gland and in ovary, brain, heart, spleen, lung, and lymph nodes. the infecting agent was identified as belonging to the genus encephalitozoon based on transmission electron microscopy and indirect immunofluorescence ...200212011264
characterization of aminopeptidase activity from three species of microsporidia: encephalitozoon cuniculi, encephalitozoon hellem, and vittaforma corneae.microsporidia are obligate intracellular parasites of the phylum microspora. to date, more than 1,200 species within 144 genera have been described, with 14 infecting humans. currently, no effective treatment exists for human microsporidiosis. in this study, the biochemical properties of the aminopeptidases were investigated within several species of microsporidia. aminopeptidase activity was detected in 3 species of microsporidia, encephalitozoon cuniculi, e. hellem, and vittaforma corneae, usi ...200212435118
sequence survey of the genome of the opportunistic microsporidian pathogen, vittaforma corneae.the microsporidian vittaforma corneae has been reported as a pathogen of the human stratum corneum, where it can cause keratitis, and is associated with systemic infections. in addition to this direct role as an infectious, etiologic agent of human disease, v. corneae has been used as a model organism for another microsporidian, enterocytozoon bieneusi, a frequent and problematic pathogen of hiv-infected patients that, unlike v. corneae, is difficult to maintain and to study in vitro. unfortunat ...200212425527
inter-strain variability of insertion/deletion events in the encephalitozoon cuniculi genome: a comparative kard-pfge analysis.we applied a two-dimensional pulsed-field gel electrophoresis procedure to the genomes of two karyotype variants assigned to two different strains of the microsporidian encephalitozoon cuniculi, termed d (strain iii) and f (strain ii). data obtained for bsshii and mlui restriction fragment length polymorphisms in each chromosome are compiled and compared to the reference strain i variant a. six insertion/deletion (indels) are found in subterminal position, some of these being characteristic of e ...200111906078
the identification of rrna maturation sites in the microsporidian encephalitozoon cuniculi argues against the full excision of presumed its1 sequence.in encephalitozoon cuniculi like in other microsporidia, the primary transcript for ssu and lsu rrnas includes only one internal transcribed spacer (its1) which separates ssu rrna from the 5.8s region associated with lsu rrna. the extraction of total rna from e. cuniculi-infected mrc5 cells using a hot phenol/chloroform procedure enabled us to perform primer extension and s1 nuclease protection experiments in the aim of identifying rrna maturation sites. our data support a simple processing (fou ...200111906080
sl-11158, a synthetic oligoamine, inhibits polyamine metabolism of encephalitozoon cuniculi. 200111906095
in vitro and in vivo evaluation of aminopeptidase inhibitors as antimicrosporidial therapies. 200111906096
encephalitozoon cuniculi infection in mice with the chronic granulomatous disease (cgd) disorder. 200111906088
isolation of encephalitozoon cuniculi using primary tissue culture techniques from a rabbit in a colony showing encephalitozoonosis.encephalitozoon spores were isolated in a primary tissue culture of the kidneys from an encephalitozoonosis-suspected rabbit in a municipal zoo in hokkaido. the isolated spores were morphologically characteristic of microsporidial ones in chromotrope stain, and proven to be e. cuniculi by a polymerase chain reaction (pcr) with a species-specific primer set and by direct dna sequencing of the pcr products.200111258462
cd8+ t lymphocytes protect scid mice against encephalitozoon cuniculi infection.microsporidia are obligate intracellular parasites that cause opportunistic infections in immunocompromised patients. the role of two main t cell subsets in anti-microsporidial immunity has been studied using an encephalitozoon cuniculi-severe combined immunodeficient (scid) mouse model. whereas scid mice reconstituted with cd4+ t lymphocyte-depleted naive balb/c splenocytes resolved the infection, adoptive transfer of cd8+ t cell-depleted splenocytes failed to protect the animals against a leth ...200111336749
immune response to encephalitozoon cuniculi infection.microsporidia are obligate intracellular parasites, which can cause complications in immunocompromised individuals. very little is known about the host immune response generated against these infectious agents. encephalitozoon cuniculi is the best studied microsporidian and the protective immune response against this parasite is mediated by cytotoxic cd8(+) t cells.200111369277
sequence and analysis of chromosome i of the amitochondriate intracellular parasite encephalitozoon cuniculi (microspora).a dna sequencing program was applied to the small (<3 mb) genome of the microsporidian encephalitozoon cuniculi, an amitochondriate eukaryotic parasite of mammals, and the sequence of the smallest chromosome was determined. the approximately 224-kb e. cuniculi chromosome i exhibits a dyad symmetry characterized by two identical 37-kb subtelomeric regions which are divergently oriented and extend just downstream of the inverted copies of an 8-kb duplicated cluster of six genes. each subtelomeric ...200111157783
microsporidian invasion apparatus: identification of a novel polar tube protein and evidence for clustering of ptp1 and ptp2 genes in three encephalitozoon species.microsporidia are unicellular eukaryotes occurring as obligate intracellular parasites which produce resistant spores. a unique motile process is represented by the sudden extrusion of the sporal polar tube for initiating entry of the parasite into a new host cell. the complete sequence of an acidic proline-rich polar tube protein (renamed ptp1) has been previously reported for encephalitozoon cuniculi and e. hellem. our immunological investigations provided evidence for an additional ptp in e. ...200111159998
the microsporidian encephalitozoon.encephalitozoon cuniculi is an attractive model system for amitochondriate intracellular eukaryotic parasites. it is characterized by a very small genome (below 3 mbp) and a unique invasion apparatus. furthermore, as an infectious agent, it is important in human and veterinary medicine. the compactness of its genome involves the reduction of rdna sequences as well as of some protein-coding genes and intergenic regions. its highly differentiated apparatus to penetrate the host cell, an extrusome- ...200111169593
[microsporidiosis].opportunistic parasites: microsporidia are primitive eukaryotic parasites widespread in a large range of animal species. these opportunistic parasites can cause infections in humans, mainly in immunocompromised patients. pathogenic species: four microsporidian species are important in human pathology, enterocytozoon bieneusi, encephalitozoon intestinalis, encephalitozoon cuniculi and encephalitozoon hellem. laboratory diagnosis: a difficult task, laboratory diagnosis is based on direct microscop ...200111225488
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