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role of interleukin (il)-18 in experimental paracoccidioidomycosis.interleukin (il)-18 has been regarded as a th1 type cytokine involved in many fungal and parasitic infections. since there have been no studies, as of yet, evaluating the role of this cytokine in paracoccidioidomycosis (pcm), we assessed the function of il-18 by using an experimental pcm model. our results showed that il-18 knockout (il-18 -/-) balb/c were more resistant to paracoccidioides brasiliensis than their littermate controls (wt). in fact, mortality rate was higher in wt mice and in the ...200818608933
recognition of laminin by paracoccidioides brasiliensis conidia: a possible mechanism of adherence to human type ii alveolar cells.this study addresses the recognition of laminin by paracoccidioides brasiliensis conidia, as well as its possible role in the adherence of conidia to a549 cells. adherence of conidia to immobilized laminin was shown to be specific, as anti-laminin antibodies, soluble laminin or the laminin-derived peptides ikvav and cdpgyigsr inhibited this interaction. rgd containing peptides and various monosaccharides had no effect on adherence, with the exception of n-acetylneuraminic acid. pre-treatment of ...200818608937
paracoccidioides brasiliensis in a liquid-based papanicolaou test from a pregnant woman: report of a case.with the exception of candida species, fungal organisms are rarely encountered in the papanicolaou test and, when encountered, usually represent contaminants rather then infections. we present the case of a healthy 29-year-old pregnant female, gravida 1, para 0, who presented for her first prenatal visit at 12-week gestation and had many large fungal-like elements with a distinct budding pattern of multiple narrow-based buds resembling a "ship's-wheel" identified in her routine liquid-based papa ...200818618723
vaccination with heat shock protein 60 induces a protective immune response against experimental paracoccidioides brasiliensis pulmonary infection.paracoccidioides brasiliensis causes a chronic granulomatous mycosis prevalent in latin america. the successful resolution of infection with this fungus is dependent on the activation of cellular immunity. we previously identified heat shock protein 60 (hsp60) as a target of the humoral response in paracoccidioidomycosis. herein we expressed the gene encoding hsp60 in escherichia coli and analyzed the immunological activity of this recombinant antigen. the immunization of balb/c mice with recomb ...200818625731
helminth infections associated with multiple sclerosis induce regulatory b cells.to assess the importance of b-cell control during parasite infections in multiple sclerosis (ms) patients.200818655096
human neutrophils susceptibility to paracoccidioides brasiliensis: an ultrastructural and cytochemical assay.paracoccidioidomycosis, caused by the dimorphic fungus paracoccidioides brasiliensis, is the most prevalent systemic mycosis of latin america, with brazil accounting for 80% of the reported cases. the great number of neutrophils found in p. brasiliensis granulomas demonstrates the importance of polymorphonuclear neutrophils (pmn) cells during this mycotic infection. it has been found that neutrophils from healthy human donors can ingest and kill the fungus through a typical phagocytic process. t ...200818404552
bacterial and fungal pneumonias after lung transplantation.the aim of this study was to evaluate the epidemiology of bacterial and fungal pneumonia in lung transplant (lt) recipients and to assess donor-to-host transmission of these microorganisms.200818455028
evidence for positive selection in putative virulence factors within the paracoccidioides brasiliensis species complex.paracoccidioides brasiliensis is a dimorphic fungus that is the causative agent of paracoccidioidomycosis, the most important prevalent systemic mycosis in latin america. recently, the existence of three genetically isolated groups in p. brasiliensis was demonstrated, enabling comparative studies of molecular evolution among p. brasiliensis lineages. thirty-two gene sequences coding for putative virulence factors were analyzed to determine whether they were under positive selection. our maximum ...200818820744
innate immunity to paracoccidioides brasiliensis infection.innate immunity is based in pre-existing elements of the immune system that directly interact with all types of microbes leading to their destruction or growth inhibition. several elements of this early defense mechanism act in concert to control initial pathogen growth and have profound effect on the adaptative immune response that further develops. although most studies in paracoccidioidomycosis have been dedicated to understand cellular and humoral immune responses, innate immunity remains po ...200818777631
comparison of transcription of multiple genes during mycelia transition to yeast cells of paracoccidioides brasiliensis reveals insights to fungal differentiation and pathogenesis.the ascomycete paracoccidioides brasiliensis is a human pathogen with a broad distribution in latin america. the infection process of p. brasiliensis is initiated by aerially dispersed mycelia propagules, which differentiate into the yeast parasitic phase in human lungs. therefore, the transition to yeast is an initial and fundamental step in the infective process. in order to identify and characterize genes involved in p. brasiliensis transition to yeast, which could be potentially associated t ...200818777633
diversity in paracoccidioides brasiliensis. the pbgp43 gene as a genetic marker.paracoccidioides brasiliensis is a temperature-dependent dimorphic fungus and the agent of paracoccidioidomycosis (pcm), which is prevalent in rural workers of latin american countries. until a decade ago, most of the studies involving p. brasiliensis used clinical isolates, since environmental samples from soil are difficult to obtain. more recently, p. brasiliensis has been isolated from infected wild and domestic animals, especially from the nine-banded armadillo dasypus novemcinctus in brazi ...200818777634
pulmonary immune responses induced in balb/c mice by paracoccidioides brasiliensis conidia.knowledge concerning the host-paracoccidioides brasiliensis interactions is abundant. yet, most of the experimental studies have used yeast cells to prepare the corresponding inoculum. as these cells do not represent the naturally infecting propagules, the corresponding experiments by-pass the earlier stages of such interactions. studies done in patients, who also harbour yeast cells, suffer from the same bias. the review presented below focuses on the immune responses of balb/c mice infected wi ...200818777636
melanin as a virulence factor of paracoccidioides brasiliensis and other dimorphic pathogenic fungi: a minireview.melanin pigments are substances produced by a broad variety of pathogenic microorganisms, including bacteria, fungi, and helminths. microbes predominantly produce melanin pigment via tyrosinases, laccases, catecholases, and the polyketide synthase pathway. in fungi, melanin is deposited in the cell wall and cytoplasm, and melanin particles ("ghosts") can be isolated from these fungi that have the same size and shape of the original cells. melanin has been reported in several human pathogenic dim ...200818777637
attempts at a peptide vaccine against paracoccidioidomycosis, adjuvant to chemotherapy.chemotherapy is the basis of treatment of paracoccidioidomycosis in its various forms. depending on the paracoccidioides brasiliensis virulence, the status of host immunity, the degree of tissue involvement and fungal dissemination, treatment can be extended for long periods with an alarming frequency of relapses. association of chemotherapy with a vaccine to boost the cellular immune response seemed a relevant project not only to reduce the time of treatment but also to prevent relapses and imp ...200818777638
international issues: meningoencephalitis due to paracoccidioides brasiliensis. 200819015480
evidence for the role of calcineurin in morphogenesis and calcium homeostasis during mycelium-to-yeast dimorphism of paracoccidioides brasiliensis.paracoccidioides brasiliensis is a dimorphic fungus that causes paracoccidioidomycosis, the most prevalent human deep mycosis in latin america. the dimorphic transition from mycelium to yeast (m-y) is triggered by a temperature shift from 25 degrees c to 37 degrees c and is critical for pathogenicity. intracellular ca(2+) levels increased in hyphae immediately after temperature-induced dimorphism. the chelation of ca(2+) with extracellular (egta) or intracellular (bapta) calcium chelators inhibi ...200818776037
the stress responsive and morphologically regulated hsp90 gene from paracoccidioides brasiliensis is essential to cell viability.paracoccidioides brasiliensis is a dimorphic fungus that causes the most prevalent systemic mycosis in latin america. the response to heat shock is involved in pathogenesis, as this pathogen switches from mycelium to yeast forms in a temperature dependent fashion that is essential to establish infection. hsp90 is a molecular chaperone that helps in the folding and stabilization of selected polypeptides. hsp90 family members have been shown to present important roles in fungi, especially in the p ...200818808717
insights into the pathobiology of paracoccidioides brasiliensis from transcriptome analysis--advances and perspectives.paracoccidioiddes brasiliensis is a thermodimorphic fungus endemic to latin america, where it causes the most prevalent systemic mycosis, paracoccidioidomycosis (pcm). dna microarray technology has been used to identify patterns of gene expression when a microbe is confronted with conditions of interest, such as in vitro and/or ex vivo interaction with specific cells. p. brasiliensis is one organism that has benefited from this approach. even though its genome has not been sequenced yet, much ha ...200818777632
lack of galectin-3 drives response to paracoccidioides brasiliensis toward a th2-biased immunity.there is recent evidence that galectin-3 participates in immunity to infections, mostly by tuning cytokine production. we studied the balance of th1/th2 responses to p. brasiliensis experimental infection in the absence of galectin-3. the intermediate resistance to the fungal infection presented by c57bl/6 mice, associated with the development of a mixed type of immunity, was replaced with susceptibility to infection and a th2-polarized immune response, in galectin-3-deficient (gal3(-/-)) mice. ...200919229338
identification of paracoccidioides brasiliensis in adrenal glands biopsies of two patients with paracoccidioidomycosis and adrenal insufficiency.the authors report two cases of adrenal insufficiency secondary to infiltration of the adrenal glands by paracoccidioides brasiliensis. the first patient had been treated for a chronic multifocal form of paracoccidiodomycosis 11 years ago. the diagnosis of the mycosis was done simultaneous with that of the adrenal insufficiency in the second patient. in both patients the diagnosis was done by direct visualization of fungus in adrenal biopsies. they were treated with hormonal supplements and itra ...200919229390
paracoccidioidomycosis in patients infected with and not infected with human immunodeficiency virus: a case-control study.epidemiologic and clinical data for 53 patients with paracoccidioidomycosis and co-infected with human immunodeficiency virus (hiv) (cases) were compared with those for 106 patients with endemic paracoccidioidomycosis (controls). the prevalence of paracoccidioides brasiliensis co-infection was estimated in 1.4% in cases of acquired immunodeficiency syndrome (aids). patients co-infected with hiv were younger, less involved in agricultural occupations; 83.7% had cd4+ cell count < 200 cells/microl. ...200919270282
detection of paracoccidioides brasiliensis gp43 gene in sputa by loop-mediated isothermal amplification method.the fungus paracoccidioides brasiliensis is the pathogen of paracoccidioidomycosis (pcm), a systemic mycosis prevalent in latin america. the loop-mediated isothermal amplification method (lamp) was used in this study to detect the presence of p. brasiliensis in sputa samples from patients with chronic pcm, suspected pcm, and a negative control. the target p. brasiliensis gp43 gene was amplified in less than 4 hr in 11 of 18 sputa samples tested. the lamp method had the advantage of speed and sim ...200919288448
bone marrow necrosis related to paracoccidioidomycosis: the first eight cases identified at autopsy.to report the first eight bone marrow necrosis (bmn) cases related to paracoccidioidomycosis (pcm) from patient autopsies with well-documented bone marrow (bm) histology and cytology.200919309401
nitric oxide participation in granulomatous response induced by paracoccidioides brasiliensis infection in mice.the role of nitric oxide (no) in granulomas of paracoccidioides brasiliensis-infected inducible no synthase-deficient c57bl/6 mice (inos ko) and their wild-type counterparts and its association with osteopontin (opn) and matrix metalloproteinases (mmps) was studied. at 15 days after infection (dai), inos ko mice showed compact and necrotic granulomas with opn+ macrophages and multinucleated giant cells, whereas wild-type mice developed loose granulomas with many fungi and opn+ cells distributed ...200919360439
molecular approaches for eco-epidemiological studies of paracoccidioides brasiliensis.medical mycology has greatly benefited from the introduction of molecular techniques. new knowledge on molecular genetics has provided both theoretical and practical frameworks, permitting important advances in our understanding of several aspects of pathogenic fungi. considering paracoccidioides brasiliensis in particular, important eco-epidemiological aspects, such as environmental distribution and new hosts were clarified through molecular approaches. these methodologies also contributed to a ...200919722090
resistance of melanized yeast cells of paracoccidioides brasiliensis to antimicrobial oxidants and inhibition of phagocytosis using carbohydrates and monoclonal antibody to cd18.paracoccidioides brasiliensis, a thermal dimorphic fungal pathogen, produces a melanin-like pigment in vitro and in vivo. we investigated the involvement of carbohydrates and monoclonal antibody to cd18, on phagocytosis inhibition, involving macrophage receptors and the resistance of melanized fungal cells to chemically generated nitric oxide (no), reactive oxygen species (ros), hypochlorite and h2o2. our results demonstrate that melanized yeast cells were more resistant than nonmelanized yeast ...200919722091
mice immunization with radioattenuated yeast cells of paracoccidiodes brasiliensis: influence of the number of immunizations.paracoccidioides brasiliensis is the agent of paracoccidioidomycosis, the most prevalent mycosis in latin america. up to the moment no vaccine has been reported. the aim of this study was to evaluate the influence of the number of immunizations on the protection elicited by radioattenuated yeast cells of p. brasiliensis. balb/c mice were divided into two groups that were immunized once (group 1) or twice (group 2), respectively. in each group, mice were divided into sub-groups that were challeng ...200919350415
increased immunoglobulin g anti-paracoccidioides brasiliensis serum antibody avidity as a predictor of favorable posttherapeutic evolution in paracoccidioidomycosis.paracoccidioidomycosis is endemic in latin america, and ca. 80% of all cases occur in brazil. little is known about antibody avidity or the evolution of such avidity in the posttherapeutic period for the different clinical presentations of the disease. in the present study, we evaluated 53 patients with paracoccidioidomycosis and calculated the avidity index. medium- and high-avidity antibodies were found in 79.5% of patients with chronic presentation (n = 39). among patients with the acute form ...200919726614
antimicrobial effect of farnesol, a candida albicans quorum sensing molecule, on paracoccidioides brasiliensis growth and morphogenesis.farnesol is a sesquiterpene alcohol produced by many organisms, and also found in several essential oils. its role as a quorum sensing molecule and as a virulence factor of candida albicans has been well described. studies revealed that farnesol affect the growth of a number of bacteria and fungi, pointing to a potential role as an antimicrobial agent.200919402910
beneficial effects of a low-protein diet on host resistance to paracoccidioides brasiliensis in mice.although protein malnutrition impairs immune functions, several studies have recently shown that protein restriction without malnutrition is beneficial to host defenses against invading pathogens and cancer. in an effort to establish the optimum diet for host resistance, we investigated the effect of different dietary protein levels on host resistance to paracoccidioides brasiliensis.200919403266
genomic dna microarray comparison of gene expression patterns in paracoccidioides brasiliensis mycelia and yeasts in vitro.paracoccidioides brasiliensis is a thermally dimorphic fungus, and causes the most prevalent systemic mycosis in latin america. infection is initiated by inhalation of conidia or mycelial fragments by the host, followed by further differentiation into the yeast form. information regarding gene expression by either form has rarely been addressed with respect to multiple time points of growth in culture. here, we report on the construction of a genomic dna microarray, covering approximately 25 % o ...200919406900
morphologic organization of pulmonary granulomas in mice infected with paracoccidioides brasiliensis.a morphologic study of the lungs was carried out in swiss mice infected with yeast isolated from paracoccidioides brasiliensis (pb18). the lung was processed 1, 2, 4, and 8 weeks after inoculation for histologic staining with hematoxylin and eosin (h&e), methenamine silver nitrate (gomori-grocott), and picrosirius to qualitative and quantitative analyses of the granulomas and the presence of fungal lesions. the numbers of cfus/g counted in the lungs were 189.8 +/- 20.64, 353.6 +/- 46.21, 547.2 + ...200919407127
inducible nitric oxide synthase-deficient mice show exacerbated inflammatory process and high production of both th1 and th2 cytokines during paracoccidioidomycosis.paracoccidioidomycosis, the major systemic mycosis in latin america, is caused by fungus paracoccidioides brasiliensis. to analyze the influence of inducible nitric oxide synthase (inos) in this disease, inos-deficient (inos(-/-)) and wild-type (wt) mice were infected intravenously with p. brasiliensis 18 isolate. we found that, unlike wt mice, inos(-/-) mice did not control fungal proliferation, and began to succumb to infection by day 50 after inoculation of yeast cells. typical inflammatory g ...200919063988
characterization and functional analysis of the beta-1,3-glucanosyltransferase 3 of the human pathogenic fungus paracoccidioides brasiliensis.the fungus paracoccidioides brasiliensis causes paracoccidioidomycosis, a systemic granulomatous mycosis prevalent in latin america. in an effort to elucidate the molecular mechanisms involved in fungus cell wall assembly and morphogenesis, beta-1,3-glucanosyltransferase 3 (pbgel3p) is presented here. pbgel3p presented functional similarity to the glucan-elongating/glycophospholipid-anchored surface/ph-regulated /essential for pseudohyphal development protein families, which are involved in fung ...200919076240
paracoccidioides brasiliensis pancreatic destruction in calomys callosus experimentally infected.the wild rodent calomys callosus is notably resistant to trypanosoma cruzi infection. in order to better characterize this animal model for experimental infections, we inoculated c. callosus intraperitoneally with paracoccidioides brasiliensis, a thermally dimorphic fungus that causes a chronic disease with severe granuloma formation in the mouse and humans. the dissemination of p. brasiliensis cells through the lungs, liver, pancreas, and spleen was assessed by histological analysis.200919422699
enolase from paracoccidioides brasiliensis: isolation and identification as a fibronectin-binding protein.paracoccidioides brasiliensis yeast cells can enter mammalian cells and may manipulate the host cell environment to favour their own growth and survival. moreover, fibronectin and several other host extracellular matrix proteins are recognized by various components of the yeast cell extracts. the present study was designed to isolate and characterize a fibronectin-binding protein from p. brasiliensis. we also compared p. brasiliensis strain 18, tested before (pb18a) and after (pb18b) animal pass ...200919429745
inhibition of human neutrophil apoptosis by paracoccidioides brasiliensis: role of interleukin-8.paracoccidioidomycosis (pcm) is a systemic mycosis caused by paracoccidiodes brasiliensis that presents a wide spectrum of clinical manifestations. because of the great number of neutrophils polymorphonuclear neutrophils (pmn) found in the p. brasiliensis granuloma, studies have been done to evaluate the role of these cells during the development of the infection. this fungus is found intracellularly in pmn and monocytes/macrophages, suggesting that it is capable of evading damage and surviving ...200919144080
amphotericin b in poly(lactic-co-glycolic acid) (plga) and dimercaptosuccinic acid (dmsa) nanoparticles against paracoccidioidomycosis.the present study reports on the preparation and testing of a desoxycholate amphotericin b (d-amb) sustained delivery system based on poly(lactic-co-glycolic acid) (plga) and dimercaptosuccinic acid (dmsa) polymeric blends (nano-d-amb) aimed at reducing the number of amb administrations required to treat mycosis.200919151037
utility of real-time pcr for the detection of paracoccidioides brasiliensis dna in the diagnosis of imported paracoccidioidomycosis.an increase in immigration from endemic regions has resulted in a number of cases of paracoccidioidomycosis (pcm) being imported into spain. a molecular diagnostic technique based on real-time pcr was developed for the detection of paracoccidioides brasiliensis dna in both culture and patients' clinical samples. a molecular beacon probe was used, labelled with fam and directed at the its1 region of ribosomic dna. the detection limit of the technique developed was 1 fg of fungal dna per microl of ...200919184775
dnahsp65 vaccination induces protection in mice against paracoccidioides brasiliensis infection.heat-shock proteins are molecules with extensive data showing their potential as immunomodulators of different types of diseases. the gene of hsp65 from mycobacterium leprae has shown prophylactic and immunotherapeutic effects against a broad arrays of experimental models including tuberculosis, leishmaniasis, arthritis and diabetes. with this in mind, we tested the dnahsp65 vaccine using an experimental model of paraccocidiodomycosis, an important endemic mycosis in latin america. the intramusc ...200919028537
curcumin as a promising antifungal of clinical interest.the antifungal activity of curcumin was evaluated against 23 fungi strains and its in vitro inhibitory effect on the adhesion of candida species to human buccal epithelial cells (bec) was also investigated.200919038979
osteopontin involvement in granuloma formation and in the severity of paracoccidioides brasiliensis infection.the participation of osteopontin (opn) in paracoccidioides brasiliensis infected mice, its association to granulomatogenesis, severity of infection, pattern of lesions, nitric oxide (no) levels and fungal load were evaluated in this investigation. immunohistochemistry analysis showed marked opn staining in extracellular matrix and in macrophages and multinucleated giant cells at the center of lesions, suggesting a possible role of opn in the distribution of these cells within the granulomas. at ...200919048431
peptides from paracoccidioides brasiliensis gp43 inhibit macrophage functions and inflammatory response.paracoccidioidomycosis (pcm) is a systemic granulomatous disease caused by paracoccidioides brasiliensis (pb), a thermal dimorphic fungus. its major antigen is a 43-kda glycoprotein. gp43 embodies different functions: it participates in evasion mechanisms during the installation of primary infection, stimulates granuloma-like formation in vitro and presents t-cell epitopes that induce protective response against the fungus. here, we investigated epitopes from gp43 inhibitory of both, macrophage ...200919026760
cross-reactivity of paracoccidioides brasiliensis, histoplasma capsulatum, and cryptococcus species in the commercial platelia aspergillus enzyme immunoassay.cross-reactivity in the platelia aspergillus enzyme immunoassay was evaluated using 120 sera from patients with paracoccidioidomycosis, histoplasmosis, and cryptococcosis. at a cutoff value of 0.5, positivity rates were 50%, 67%, and 50%, respectively. the implications for these findings are discussed.200919020109
paracoccidioides brasiliensis vaccine formulations based on the gp43-derived p10 sequence and the salmonella enterica flic flagellin.paracoccidioidomycosis (pcm) is a systemic granulomatous disease caused by the dimorphic fungus paracoccidioides brasiliensis. anti-pcm vaccine formulations based on the secreted fungal cell wall protein (gp43) or the derived p10 sequence containing a cd4(+) t-cell-specific epitope have shown promising results. in the present study, we evaluated new anti-pcm vaccine formulations based on the intranasal administration of p. brasiliensis gp43 or the p10 peptide in combination with the salmonella e ...200919204092
interleukin-6 treatment enhances human monocyte permissiveness for paracoccidioides brasiliensis growth by modulating cytokine production.the effect of interleukin (il)-6 on cytokine production was evaluated in human monocyte cultures infected with the virulent strain of paracoccidioides brasiliensis (pb18). peripheral blood monocytes from healthy individuals were preincubated for 24 h with or without human recombinant il-6, and then challenged with pb18 for 4 h and 18 h. p. brasiliensis growth was assessed by viable fungi recovery from co-cultures after plating on bhi-agar. moreover, tumor necrosis factor-alpha (tnf-alpha), il-6 ...200918668419
transcription regulation of the pbgp43 gene by nitrogen in the human pathogen paracoccidioides brasiliensis.we show indirect evidences for the possible involvement of nit2-like binding motifs in transcription modulation of the pbgp43 gene, which codes for an important antigen from the human fungal pathogen paracoccidioides brasiliensis. this investigation was motivated by the finding of 23 nit2-like sites within the proximal -2047 nucleotides of the pbgp43 5' intergenic region from the pb339 isolate. they compose four clusters, two of them identical. we found four nit2-containing probes that were posi ...200919013535
psoas abscesses caused by paracoccidioides brasiliensis in an adolescent.in order to describe the case of an adolescent who developed psoas abscesses caused by paracoccidiodes brasiliensis.200918781395
the malate synthase of paracoccidioides brasiliensis is a linked surface protein that behaves as an anchorless adhesin.the pathogenic fungus paracoccidioides brasiliensis is the agent of paracoccidioidomycosis (pcm). this is a pulmonary mycosis acquired by inhalation of fungal airborne propagules that can disseminate to several organs and tissues leading to a severe form of the disease. adhesion and invasion to host cells are essential steps involved in the internalization and dissemination of pathogens. inside the host, p. brasiliensis may use the glyoxylate cycle for intracellular survival.200920034376
polymorphism in the flanking regions of the pbgp43 gene from the human pathogen paracoccidioides brasiliensis: search for protein binding sequences and poly(a) cleavage sites.paracoccidioides brasiliensis is a thermo-dimorphic fungus that causes paracoccidiodomycosis (pcm). glycoprotein gp43 is the fungal main diagnostic antigen, which can also protect against murine pcm and interact with extracellular matrix proteins. it is structurally related to glucanases, however not active, and whose expression varies considerably. we have presently studied polymorphisms in the pbgp43 flanking regions to help understand such variations.200920042084
climate and acute/subacute paracoccidioidomycosis in a hyper-endemic area in brazil.paracoccidioidomycosis (pcm) is latin america's most prevalent systemic mycosis, carrying an important social burden. its agent, paracoccidioides brasiliensis, has rarely been identified in nature. studies characterizing acute/subacute pcm incidence and their relationship with climate variables are not available. this work analysed a series of acute/subacute cases that occurred in the botucatu area, são paulo state, brazil, from 1969 to 1999, as an outcome of weather variability.200919433518
molecular phylogeny of animal pathogen lacazia loboi inferred from rdna and dna coding sequences.lacazia loboi is a geographically restricted, uncultivated fungal pathogen of humans and dolphins. previous investigations using 18s small unit rdna, chitin synthase 2 and gp43 dna sequences positioned l. loboi as a close relative of paracoccidioides brasiliensis. however, given the few individuals of l. loboi studied and the high degree of genetic variation observed in p. brasiliensis, the existence of l. loboi as an independent species has been questioned. to investigate the phylogenetic posit ...200919439181
identification and characterization of antigenic proteins potentially expressed during the infectious process of paracoccidioides brasiliensis.paracoccidioides brasiliensis causes paracoccidioidomycosis (pcm), a systemic mycosis presenting clinical manifestations ranging from mild to severe forms. a p. brasiliensis cdna expression library was produced and screened with pooled sera from pcm patients adsorbed against antigens derived from in vitro-grown p. brasiliensis yeast cells. sequencing dna inserts from clones reactive with pcm patients sera indicated 35 open reading frames presenting homology to genes involved in metabolic pathway ...200919500685
preferential transcription of paracoccidioides brasiliensis genes: host niche and time-dependent expression.paracoccidioides brasiliensis causes infection through inhalation by the host of airborne propagules from the mycelium phase of the fungus. this fungus reaches the lungs, differentiates into the yeast form and is then disseminated to virtually all parts of the body. here we review the identification of differentially-expressed genes in host-interaction conditions. these genes were identified by analyzing expressed sequence tags (ests) from p. brasiliensis cdna libraries. the p. brasiliensis was ...200919547877
inteins in pathogenic fungi: a phylogenetic tool and perspectives for therapeutic applications.inteins or 'internal proteins' are coding sequences that are transcribed and translated with flanking sequences (exteins). after translation, the inteins are excised by an autocatalytic process and the host protein assumes its normal conformation and develops its expected function. these parasitic genetic elements have been found in important, conserved proteins in all three domains of life. most of the eukaryotic inteins are present in the fungi kingdom and the prp8 intein is one of the most wi ...200919547879
tlr2 is a negative regulator of th17 cells and tissue pathology in a pulmonary model of fungal infection.to study the role of tlr2 in a experimental model of chronic pulmonary infection, tlr2-deficient and wild-type mice were intratracheally infected with paracoccidioides brasiliensis, a primary fungal pathogen. compared with control, tlr2(-/-) mice developed a less severe pulmonary infection and decreased no synthesis. equivalent results were detected with in vitro-infected macrophages. unexpectedly, despite the differences in fungal loads both mouse strains showed equivalent survival times and se ...200919553529
interferon-gamma and interleukin-4 single nucleotide gene polymorphisms in paracoccidioidomycosis.the gene polymorphisms interferon-gamma (ifn-gamma) +874 t/a and interleukin (il)-4 -590 c/t have been associated with the altered production of cytokines. therefore, they might be indicative of the occurrence of paracoccidioidomycosis (pcm) caused by paracoccidioides brasiliensis. the analysis of single nucleotide polymorphism (snp) at position+874 ifn-gamma showed an increase occurrence of a/t genotype in both pcm patients and healthy individuals as control (hic) (56% and 45%, respectively), w ...200919682920
transcription levels of chs5 and chs4 genes in paracoccidioides brasiliensis mycelial phase, respond to alterations in external osmolarity, oxidative stress and glucose concentration.the complete sequence of paracoccidioides brasiliensis chs5 gene, encoding a putative chitin synthase revealed a 5583nt open reading frame, interrupted by three introns of 82, 87 and 97bp (genbank accession no ef654132). the deduced protein contains 1861 amino acids with a predicted molecular weight of 206.9kda. both its large size and the presence of a n-terminal region of approx. 800 residues with a characteristic putative myosin motor-like domain, allow us to include pbrchs5 into class v fung ...200919616626
cdc42p controls yeast-cell shape and virulence of paracoccidioides brasiliensis.paracoccidioides brasiliensis is characterized by a multiple budding phenotype and a polymorphic cell growth, leading to the formation of cells with extreme variations in shape and size. since cdc42 is a pivotal molecule in establishing and maintaining polarized growth for diverse cell types, as well as during pathogenesis of certain fungi, we evaluated its role during cell growth and virulence of the yeast-form of p. brasiliensis. we used antisense technology to knock-down pbcdc42's expression ...200919686860
screening non-coding rnas in transcriptomes from neglected species using portrait: case study of the pathogenic fungus paracoccidioides brasiliensis.transcriptome sequences provide a complement to structural genomic information and provide snapshots of an organism's transcriptional profile. such sequences also represent an alternative method for characterizing neglected species that are not expected to undergo whole-genome sequencing. one difficulty for transcriptome sequencing of these organisms is the low quality of reads and incomplete coverage of transcripts, both of which compromise further bioinformatics analyses. another complicating ...200919653905
induction of apoptosis in a549 pulmonary cells by two paracoccidioides brasiliensis samples.paracoccidioidomycosis presents a variety of clinical manifestations and paracoccidioides brasiliensis can reach many tissues, most importantly the lungs. the ability of the pathogen to interact with host surface structures is essential to its virulence. the interaction between p. brasiliensis and epithelial cells has been studied, with particular emphasis on the induction of apoptosis. to investigate the expression of different apoptosis-inducing pathways in human a549 cells, we infected these ...200919820837
effect of dietary oils on host resistance to fungal infection in psychologically stressed mice.psychological stress can modulate host defense against invading pathogens. in this study, we investigated the effect of dietary oils on social isolation stress-induced modulation of host resistance to paracoccidioides brasiliensis. in olive oil-fed mice, 3 weeks of isolation stress resulted in temporarily delayed clearance of this fungus in the liver compared with group-housed mice. by contrast, in soybean oil-fed mice, isolation stress had no significant effect on antifungal activity. the olive ...200919734677
matrix metalloproteinases with gelatinolytic activity induced by paracoccidioides brasiliensis infection.matrix metalloproteinases (mmps) modulate extracellular matrix turnover, inflammation and immunity. we studied mmp-9 and mmp-2 in experimental paracoccidioidomycosis. at 15 and 120 days after infection (dai) with virulent paracoccidioides brasiliensis, mmp-9 was positive by immunohistochemistry in multinucleated giant cells, in mononuclear cells with macrophage and lymphocyte morphologies and also in fungal cells in the lesions of susceptible and resistant mice. using gelatin zymography, pro- an ...200919765107
the malate synthase of paracoccidioides brasiliensis pb01 is required in the glyoxylate cycle and in the allantoin degradation pathway.in the present study, we examined the characteristics of cdna, the regulation of the gene expression of paracoccidioides brasiliensis mls (pbmls), and the enzymatic activity of the protein p. brasiliensis mls (pbmls) from the p. brasiliensis pb01 isolate. pbmls cdna contains 1617 bp, encoding a protein of 539 amino acids with a predicted molecular mass of 60 kda. the protein presents the mlss family signature, the catalytic residues essential for enzymatic activity and the peroxisomal/glyoxysoma ...200919888806
importance of xenarthrans in the eco-epidemiology of paracoccidioides brasiliensis.several pathogens that cause important zoonotic diseases have been frequently associated with armadillos and other xenarthrans. this mammal group typically has evolved on the south american continent and many of its extant species are seriously threatened with extinction. natural infection of armadillos with paracoccidioides brasiliensis in hyperendemic areas has provided a valuable opportunity for understanding the role of this mammal in the eco-epidemiology of paracoccidioidomycosis (pcm), one ...200919919716
detection of a homotetrameric structure and protein-protein interactions of paracoccidioides brasiliensis formamidase lead to new functional insights.abstract paracoccidioides brasiliensis causes paracoccidioidomycosis, a systemic mycosis in latin america. formamidases hydrolyze formamide, putatively plays a role in fungal nitrogen metabolism. an abundant 45-kda protein was identified as the p. brasiliensis formamidase. in this study, recombinant formamidase was overexpressed in bacteria and a polyclonal antibody to this protein was produced. we identified a 180-kda protein species reactive to the antibody produced in mice against the p. bras ...200920002196
characterization of a secreted aspartyl protease of the fungal pathogen paracoccidioides brasiliensis.paracoccidioides brasiliensis is a thermally dimorphic fungus that causes paracoccidioidomycosis, a human systemic disease prevalent in latin america. proteases have been described as playing an important role in the host invasion process in many pathogenic microorganisms. here we describe the identification and characterization of a secreted aspartyl protease (pbsap), isolated from a cdna library constructed with rnas of mycelia transitioning to yeast cells. recombinant pbsap was produced in es ...200920028235
tlr-2, tlr-4 and dectin-1 expression in human monocytes and neutrophils stimulated by paracoccidioides brasiliensis.paracoccidioidomycosis (pcm) is an endemic mycosis in latin america caused by the dimorphic fungus paracoccidioides brasiliensis. the pattern of the immune responses to p. brasiliensis determines the disease progression and clinical outcome. innate immune response is mediated by phagocytic cells, such as macrophage and neutrophils, which ingest and kill invading pathogens and then trigger the adaptive immune system through the secretion of cytokines and chemokines. the c-type like lectin recepto ...200919888805
acute disseminated paracoccidioidomycosis in a 3 year-old child.clinical cases involving paracoccidioidomycosis in children, diagnosed in mato grosso state, in the central western region of brazil, are rare despite the state being classified with a moderate to high incidence. we describe a clinical case of infant acute disseminated paracoccidioidomycosis in mato grosso state, brazil, highlighting the importance of early differential diagnosis from other severe pathologies, thus contributing to the survival of compromised patients. a 3 year-old male child, we ...200920191206
chapter 6--paracoccidioidomycosis.paracoccidioidomycosis is a systemic mycosis caused by the dimorphic fungus paracoccidioides brasiliensis. the disease is restricted to latin america. it is the principal systemic mycosis in brazil, with higher incidences in the southern, southeastern and central regions. the disease is acquired by inhaling fungal propagules. in endemic areas, the primary infection occurs during childhood and involves the immune system. the most common chronic form of paracoccidioidomycosis in adults is the mult ...200920126928
primary complex of paracoccidioidomycosis and hypereosinophilia.primary infection with paracoccidioides brasiliensis has rarely been observed. a 28-year-old male patient presented with a three-month history of fever, respiratory symptoms and malaise. chest x-rays revealed bilateral apical infiltrates, right pleuritis and hilar lymphadenomegaly. the patient presented with leukocytosis, severe eosinophilia and increasing titers of anti-p. brasiliensis antibodies in serum. to our knowledge, this is the first report of the primary pulmonary lymph node complex of ...200920126931
[identification of genes associated with germination of conidia to form mycelia in the fungus paracoccidioides brasiliensis].paracoccidioides brasiliensis is a thermo-dimorphic fungus. at room temperature it grows as a mold that produces conidia, whereas in the vertebrate host it grows as a multiple-budding yeast. the molecular mechanisms involved in the germination from the conidia to the mycelia process remain unknown.200920436992
[systemic mycosis: factors associated with death among patients infected with the human immunodeficiency virus, cuiabá, state of mato grosso, brazil, 2005-2008].between 2005 and 2008, the prevalence of systemic mycosis among 1,300 hiv/aids patients in cuiabá, mato grosso, was 4.6%. the fungus species isolated were cryptococcus neoformans in 50%, cryptococcus gattii in 1.6%, cryptococcus spp in 6.6%, histoplasma capsulatum in 38.3% and paracoccidioides brasiliensis in 3.3%. death was recorded in the cases of 32 patients (53.3%), and cryptococcosis was the main cause. the cd4+ t lymphocyte count was low and similar among patients who survived or died due ...200920209358
activation of the alternative complement pathway in canine normal serum by paracoccidioides brasiliensis.the dimorphic fungus paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a human granulomatous disease. recently the first case of natural disease in dogs was reported. the complement system is an important effector component of humoral immunity against infectious agents. therefore, the aim of this study was to evaluate the activation of the dog alternative complement pathway by p. brasiliensis. initially, the ability of erythrocytes of guinea pig, rabbit, sheep, ch ...200924031350
disseminated paracoccidioidomycosis with a single brainstem lesion. a case report and literature review.paracoccidioidomycosis is a systemic disease endemic to subtropical areas in central and south america caused by a dimorphic fungus known as paracoccidioides brasiliensis. central nervous system involvement is a severe complication of the systemic disease, and has been found in approximately 13% of patients. this paper describes the case of a patient whose computed tomography scan and magnetic resonance imaging showed a single tumor-like lesion in the brainstem. histopathological and mycological ...201024148639
first report on the diagnosis and treatment of encephalic and urinary paracoccidioidomycosis in a cat.a male persian cat was presented with persistent fever, anorexia, weakness, hypopyon, nystagmus, and intention tremors. the hemogram showed severe neutropenia and laboratory analysis on cerebrospinal fluid (csf) smears revealed abundant yeast cells compatible with paracoccidioides brasiliensis. urinalysis demonstrated persistent funguria and an increased urine protein-to-creatinine ratio (upc) in addition to mild azotemia. long-term therapy with oral fluconazole was effective in controlling the ...201020471880
disseminated paracoccidioidomycosis in a southern two-toed sloth (choloepus didactylus).a southern two-toed sloth (choloepus didactylus), originally acquired from french guiana, died while maintained in quarantine in a pet store in monterrey, mexico. large yeast cells with multiple buds compatible with paracoccidioides brasiliensis were observed in disseminated granulomatous lesions in the lungs, liver, spleen and kidney. transmission electron microscopical examination supported the diagnosis. this is the first report of paracoccidioidomycosis in a two-toed sloth.201020961559
mannose-binding lectin complement pathway plays a key role in complement activation by paracoccidioides brasiliensis.paracoccidioides brasiliensis (pb) is a dimorphic fungal pathogen that causes paracoccidioidomycosis, the most severe deep mycosis from south america. although cell mediated immunity is considered the most efficient protective mechanism against pb infection, mechanisms of innate immunity are poorly defined. herein, we investigated the interaction of the complement system with high and low virulence isolates of pb. we demonstrated that pb18, a high virulence pb isolate, when incubated with normal ...201021035191
a semi-nested pcr assay for molecular detection of paracoccidioides brasiliensis in tissue samples.paracoccidioidomycosis is a systemic infection caused by paracoccidioides brasiliensis.201021181033
involvement of an alternative oxidase in oxidative stress and mycelium-to-yeast differentiation in paracoccidioides brasiliensis.paracoccidioides brasiliensis is a thermodimorphic human pathogenic fungus that causes paracoccidioidomycosis (pcm), which is the most prevalent systemic mycosis in latin america. differentiation from the mycelial to the yeast form (m-to-y) is an essential step for the establishment of pcm. we evaluated the involvement of mitochondria and intracellular oxidative stress in m-to-y differentiation. m-to-y transition was delayed by the inhibition of mitochondrial complexes iii and iv or alternative ...201021183691
response of paracoccidioides brasiliensis pb01 to stressor agents and cell wall osmoregulators.in its attempt to survive, the fungal cell can change the cell wall composition and/or structure in response to environmental stress. the molecules involved in these compensatory mechanisms are a possible target for the development of effective antifungal agents. in the thermodimorphic fungus paracoccidioides brasiliensis pb01, the main polymers that compose the cell wall are chitin and glucans. these polymers form a primary barrier that is responsible for the structural integrity and formation ...201021215956
protein-losing enteropathy in paracoccidioidomycosis identified by scintigraphy: report of three cases.paracoccidioidomycosis (pcm) or south american blastomycosis is a systemic fungal infection caused by a dimorphic fungus, paracoccidioides brasiliensis. it represents the most important systemic mycosis in south america and with higher prevalence among male inhabitants of the rural area. pcm usually affects the lungs, and rarely the intestines. the authors report three cases of pcm, treated at the university hospital of universidade federal de mato grosso do sul whose scintigraphy with technetiu ...201021221488
long-term visual safety of voriconazole in adult patients with paracoccidioidomycosis.voriconazole is an antifungal agent with in vitro activity and clinical efficacy against yeasts, molds, and dimorphic fungi (eg, paracoccidioides brasiliensis). the safety profile of voriconazole includes transient visual adverse events (vaes) that resolve while undergoing treatment or after its discontinuation.201021316537
phenotypic and functional characterization of pulmonary macrophages subpopulations after intratracheal injection of paracoccidioides brasiliensis cell wall components.a shift in the activation of pulmonary macrophages characterized by an increase of il-1, tnf-α and il-6 production has been induced in mice infected with paracoccidioides brasiliensis. it is still unclear whether a functional shift in the resident alveolar macrophage population would be responsible for these observations due to the expression of cell surface molecules. we investigated pulmonary macrophages by flow cytometry from mice treated with p. brasiliensis derivatives by intratracheal rout ...201021257226
oral exfoliative cytology for the diagnosis of paracoccidoidomycosis in a patient with human immunodeficiency virus: a case report.paracoccidioidomycosis is not the most common fungal disease in patients infected with human immunodeficiency virus (hiv), except for endemic regions in latin america countries.201021428160
[paracoccidioidomicosis and multidrug-resistant tuberculosis (tbc-mdr) in patient coinfected with hiv and hepatitis c].a case of an adult male patient diagnosed with hiv and hepatitis c co infection is presented. he had granu-lomatuos hepatitis and blood smear positive to paracoccidioides brasiliensis concomitant to the detection of mdr mycobacterium tuberculosis in sputum further complicated with reactivation of cytomegalovirus (possible pancreatitis and retinitis). difficulties in diagnostic and therapeutic approach in a patient with multiple infections are reviewed.201021279295
poly(lactic acid-glycolic acid) nanoparticles markedly improve immunological protection provided by peptide p10 against murine paracoccidioidomycosis.background and purpose: the present study reports on the preparation and testing of a sustained delivery system for the immunomodulatory peptide p10 aimed at reducing the in vivo degradation of the peptide and the amount required to elicit a protective immune response against paracoccidioidomycosis. experimental approach: balb/c mice were infected with the yeast paracoccidioides brasiliensis to mimic the chronic form of paracoccidioidomycosis. the animals were treated daily with sulfamethoxazole ...201020136827
paracoccidioidomycosis in a spanish missionary.paracoccidioidomycosis is the most important systemic mycosis in south america. in europe the disease is very rare and only found in returning travelers. here we report on a 56-year-old spanish missionary with respiratory symptoms but no other affected systems. diagnosis was made based on serology and pcr for paracoccidioides brasiliensis.201020412183
effects of a high-protein diet on host resistance to paracoccidioides brasiliensis in mice.we investigated the effects of high protein intake on host resistance to paracoccidioides brasiliensis. two-d fasted mice were infected with p. brasiliensis and refed on diets with three different levels (54%, 20%, and 5%) of casein. the mice refed the 54% casein diet showed reduced antifungal activity in the spleen and liver as compared with the mice refed the 5% or the 20% casein diet. after infection, increases in spleen and liver mrna levels of myeloperoxidase, cathepsin-g, and elastase-2 we ...201020208341
simultaneous infection of human host with genetically distinct isolates of paracoccidioides brasiliensis.this study is the first report on genetic differences between isolates of paracoccidioides brasiliensis from a single patient. we describe a simultaneous infection with genetically distinct isolates of p. brasiliensis in a patient with chronic paracoccidioidomycosis. the clinical isolates were obtained from lesions in different anatomical sites and were characterised by random amplified polymorphic dna (rapd) analysis. the rapd technique can be helpful for distinguishing between clinical isolate ...201020209331
immunological basis for the gender differences in murine paracoccidioides brasiliensis infection.this study aimed to investigate the immunological mechanisms involved in the gender distinct incidence of paracoccidioidomycosis (pcm), an endemic systemic mycosis in latin america, which is at least 10 times more frequent in men than in women. then, we compared the immune response of male and female mice to paracoccidioides brasiliensis infection, as well as the influence in the gender differences exerted by paracoccin, a p. brasiliensis component with carbohydrate recognition property. high pr ...201020505765
toll-like receptor 4 signaling leads to severe fungal infection associated with enhanced proinflammatory immunity and impaired expansion of regulatory t cells.toll-like receptors (tlrs) present in innate immune cells recognize pathogen molecular patterns and influence immunity to control the host-parasite interaction. the objective of this study was to characterize the involvement of tlr4 in the innate and adaptive immunity to paracoccidioides brasiliensis, the most important primary fungal pathogen of latin america. we compared the responses of c3h/hej mice, which are naturally defective in tlr4 signaling, with those of c3h/hepas mice, which express ...201020008536
tgf-beta and cd23 are involved in nitric oxide production by pulmonary macrophages activated by beta-glucan from paracoccidioides brasiliensis.pulmonary macrophages (pm), which are cd11b/cd18(+) and cd23(+), may be involved in the onset of inflammatory events caused by paracoccidioides brasiliensis in the lungs. in the present study, we measured the nitric oxide (no) and interleukin in pm production after intratracheal (i.t.) inoculation of an enriched beta-glucan cell wall fraction from p. brasiliensis (fraction f1). balb/c and c57/bl6 (b6) mice were i.t. treated with fraction f1, and their pm were restimulated in vitro with lps and i ...201019949959
expression of paracoccidioides brasiliensis chs3 in a saccharomyces cerevisiae chs3 null mutant demonstrates its functionality as a chitin synthase gene.we report the isolation and sequencing of chs3, a gene that encodes one of several chitin synthases in paracoccidioides brasiliensis, a medically important fungus restricted geographically to latin america. the gene contains a single open reading frame of 3817 bp with two introns (71 and 86 bp) and encodes a 1220 amino acid polypeptide with high similarity to other fungal chitin synthases. northern analysis reveals a high expression of chs3 in the pathogenic yeast-like phase of the fungus and at ...201020037924
hsp65 dna as therapeutic strategy to treat experimental paracoccidioidomycosis.the conventional treatment for paracoccidioidomycosis, the most prevalent mycosis in latin america, involves long periods of therapy resulting in sequels and high frequency of relapses. the search for new alternatives of treatment is necessary. previously, we have demonstrated that the hsp65 gene from mycobacterium leprae shows prophylactic effects against murine paracoccidioidomycosis. here, we tested the dnahsp65 immunotherapy in balb/c mice infected with paracoccidioides brasiliensis, the age ...201020045500
[endemic fungal pathogens in a rural setting of argentina: seroepidemiological study in dogs].three fungal species causing human disease, namely paracoccidioides brasiliensis, histoplasma capsulatum and coccidioides sp., are endemic in different areas of argentina. rates of infection in domestic dogs have been used in other latin american countries as indicators of the presence of these pathogens in a given area. we used such an approach to investigate the epidemiological relevance of paracoccidiodomycosis, histoplasmosis and coccidioidomycosis in our country.201020097588
effect of anti-glycosphingolipid monoclonal antibodies in pathogenic fungal growth and differentiation. characterization of monoclonal antibody mest-3 directed to manpalpha1-->3manpalpha1-->2ipc.studies carried out during the 1990's demonstrated the presence of fungal glycoinositol phosphorylceramides (gipcs) with unique structures, some of them showed reactivity with sera of patients with histoplasmosis, paracoccidioidomycosis or aspergillosis. it was also observed that fungal gipcs were able to inhibit t lymphocyte proliferation "in vitro", and studies regarding the importance of these molecules to fungal survival showed that many species of fungi are vulnerable to inhibitors of sphin ...201020156351
expression of activation and cytotoxic molecules by peripheral blood lymphocytes of patients with paracoccidioidomycosis.in a previous study, we reported an increased number of t cd8(+) cells in the bronchoalveolar lavage (bal) of patients with pulmonary paracoccidioidomycosis, suggesting a role for these cells in the local immune response. the aims of this study were to verify, by flow cytometry, the activation state, as well as the production of cytotoxic molecules by peripheral blood lymphocytes (cd8(+) and cd4(+)). specimens were obtained from patients with paracoccidioidomycosis (pcm), individuals with pcm-in ...201020163282
identification and characterization of tc1/mariner-like dna transposons in genomes of the pathogenic fungi of the paracoccidioides species complex.paracoccidioides brasiliensis (eukaryota, fungi, ascomycota) is a thermodimorphic fungus, the etiological agent of paracoccidioidomycosis, the most important systemic mycoses in latin america. three isolates corresponding to distinct phylogenetic lineages of the paracoccidioides species complex had their genomes sequenced. in this study the identification and characterization of class ii transposable elements in the genomes of these fungi was carried out.201020178623
purification of paracoccidioides brasiliensis catalase p: subsequent kinetic and stability studies.catalases are essential components of the cellular equipment to cope with oxidative stress. here we have purified a highly abundant catalase p of paracoccidioides brasiliensis (pbcatp) that is preferentially expressed in the parasitic yeast phase. this oxidative stress-induced protein was isolated from yeast cells grown in the presence of 15 mm of hydrogen peroxide (h(2)o(2)). we have used consecutive steps of protein precipitation and gel filtration chromatography to achieve the purified protei ...201019897569
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