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1.
Int J Mol Sci ; 23(3)2022 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-35162991

RESUMO

Malaria parasites require multiple phosphorylation and dephosphorylation steps to drive signaling pathways for proper differentiation and transformation. Several protein phosphatases, including protein phosphatase 1 (PP1), one of the main dephosphorylation enzymes, have been shown to be indispensable for the Plasmodium life cycle. The catalytic subunit of PP1 (PP1c) participates in cellular processes via dynamic interactions with a vast number of binding partners that contribute to its diversity of action. In this study, we used Plasmodium berghei transgenic parasite strains stably expressing PP1c or its inhibitor 2 (I2) tagged with mCherry, combined with the mCherry affinity pulldown of proteins from asexual and sexual stages, followed by mass spectrometry analyses. Mapped proteins were used to identify interactomes and to cluster functionally related proteins. Our findings confirm previously known physical interactions of PP1c and reveal enrichment of common biological processes linked to cellular component assembly in both schizonts and gametocytes to biosynthetic processes/translation in schizonts and to protein transport exclusively in gametocytes. Further, our analysis of PP1c and I2 interactomes revealed that nuclear export mediator factor and peptidyl-prolyl cis-trans isomerase, suggested to be essential in P. falciparum, could be potential targets of the complex PP1c/I2 in both asexual and sexual stages. Our study emphasizes the adaptability of Plasmodium PP1 and provides a fundamental study of the protein interaction landscapes involved in a myriad of events in Plasmodium, suggesting why it is crucial to the parasite and a source for alternative therapeutic strategies.


Assuntos
Malária/parasitologia , Plasmodium berghei/fisiologia , Proteína Fosfatase 1/metabolismo , Proteínas/metabolismo , Proteômica/métodos , Animais , Sítios de Ligação , Cromatografia Líquida , Estágios do Ciclo de Vida , Masculino , Camundongos , Organismos Geneticamente Modificados , Plasmodium berghei/patogenicidade , Domínios Proteicos , Mapas de Interação de Proteínas , Proteína Fosfatase 1/genética , Proteínas/genética , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Espectrometria de Massas em Tandem
2.
Med Mycol ; 60(1)2021 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-34734270

RESUMO

Pneumocystis jirovecii colonization is frequent during chronic obstructive pulmonary disease (COPD) and patients constitute potential contributors to its interhuman circulation. However, the existence of an environmental reservoir cannot be excluded. We assessed the prevalence and factors associated with Pneumocystis colonization during COPD, and studied circulation between patients and their domestic environment. Pneumocystis molecular detection and mtLSU genotyping were performed in oro-pharyngeal washes (OPW) sampled in 58 patients with COPD acute exacerbation, and in indoor dust, sampled in patients' homes using electrostatic dust collectors (EDCs). Lung and systemic inflammation was assessed. Pneumocystis carriage was evaluated in 28 patients after 18 months at stable state. Pneumocystis was detected in 11/58 OPWs during exacerbation (19.0%). Colonized patients presented a significantly lower body mass index, and higher serum IL-17 and CD62P. One patient presented positive detection of typable isolates in both OPW and EDC, with both isolates harboring mtLSU genotype 3. Pneumocystis genotype 1 was further detected in EDCs from three non-colonized patients and one colonized patient with non-typable isolate. Genotypes 1 and 2 were predominant in clinical isolates (both 42%), with genotype 3 representing 16% of isolates. Pneumocystis was detected in 3/28 patients at stable state (10.7%). These data suggest that Pneumocystis colonization could be facilitated by a lower BMI and be related to acute alteration of lung function during COPD exacerbation. It also suggests Th17 pathway and platelet activation could be involved in the anti-Pneumocystis response during colonization. Last, Pneumocystis detection in EDCs supports its potential persistence in indoor dust. LAY SUMMARY: Chronic obstructive pulmonary disease patients tend to be more frequently colonized by Pneumocystis during exacerbation (19.0%) than at stable state (10.7%). Factors associated with colonization include lower BMI, higher IL-17, and CD62P. Pneumocystis detection in patients' dwellings suggests potential persistence in indoor dust.


Assuntos
Pneumocystis carinii , Pneumonia por Pneumocystis , Doença Pulmonar Obstrutiva Crônica , Genótipo , Ambiente Domiciliar , Humanos , Doença Pulmonar Obstrutiva Crônica/complicações
3.
Med Mycol ; 55(5): 568-572, 2017 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-27811179

RESUMO

In a prospective bicentric study, Pneumocystis jirovecii excretion and diffusion was explored in air samples collected in the rooms occupied by 17 Pneumocystis-colonized patients. P. jirovecii DNA was detected by real-time PCR in the air collected from 3 patients' rooms (17.6%), with identical genotypes in corresponding clinical and air samples. Pneumocystis DNA was detected for 2/3 patients with autoimmune disease treated with corticosteroids versus 1/6 patients with hematologic disease and 0/5 kidney transplant recipients. These data confirm the possible excretion of the fungus by Pneumocystis-colonized patients and thus bring additional arguments for the prevention of airborne transmission in hospital wards.


Assuntos
Microbiologia do Ar , Infecção Hospitalar/transmissão , Pneumocystis carinii/isolamento & purificação , Pneumocystis carinii/fisiologia , Pneumonia por Pneumocystis/microbiologia , Pneumonia por Pneumocystis/transmissão , Adulto , Idoso , Infecção Hospitalar/microbiologia , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , Feminino , Genótipo , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Adulto Jovem
4.
Respir Res ; 17(1): 66, 2016 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-27250970

RESUMO

Pulmonary surfactant is a complex fluid that comprises phospholipids and four proteins (SP-A, SP-B, SP-C, and SP-D) with different biological functions. SP-B, SP-C, and SP-D are essential for the lungs' surface tension function and for the organization, stability and metabolism of lung parenchyma. SP-A and SP-D, which are also known as pulmonary collectins, have an important function in the host's lung immune response; they act as opsonins for different pathogens via a C-terminal carbohydrate recognition domain and enhance the attachment to phagocytic cells or show their own microbicidal activity by increasing the cellular membrane permeability. Interactions between the pulmonary collectins and bacteria or viruses have been extensively studied, but this is not the same for fungal pathogens. SP-A and SP-D bind glucan and mannose residues from fungal cell wall, but there is still a lack of information on their binding to other fungal carbohydrate residues. In addition, both their relation with immune cells for the clearance of these pathogens and the role of surfactant proteins' regulation during respiratory fungal infections remain unknown. Here we highlight the relevant findings associated with SP-A and SP-D in those respiratory mycoses where the fungal infective propagules reach the lungs by the airways.


Assuntos
Pneumopatias Fúngicas/metabolismo , Pulmão/metabolismo , Pneumonia/metabolismo , Proteína A Associada a Surfactante Pulmonar/metabolismo , Proteína D Associada a Surfactante Pulmonar/metabolismo , Animais , Citocinas/imunologia , Citocinas/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Mediadores da Inflamação/imunologia , Mediadores da Inflamação/metabolismo , Pulmão/imunologia , Pulmão/microbiologia , Pneumopatias Fúngicas/imunologia , Pneumopatias Fúngicas/microbiologia , Pneumonia/imunologia , Pneumonia/microbiologia , Proteína A Associada a Surfactante Pulmonar/imunologia , Proteína D Associada a Surfactante Pulmonar/imunologia
5.
J Eukaryot Microbiol ; 63(3): 309-17, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-26509699

RESUMO

To quantitatively assess the risk of contamination by Pneumocystis depending on the degree of immunosuppression (ID) of the exposed rat hosts, we developed an animal model, where rats went through different doses of dexamethasone. Then, natural and aerial transmission of Pneumocystis carinii occurred during cohousing of the rats undergoing gradual ID levels (receivers) with nude rats developing pneumocystosis (seeders). Following contact between receiver and seeder rats, the P. carinii burden of receiver rats was determined by toluidine blue ortho staining and by qPCR targeting the dhfr monocopy gene of this fungus. In this rat model, the level of circulating CD4(+) and CD8(+) T lymphocytes remained significantly stable and different for each dose of dexamethasone tested, thus reaching the goal of a new stable and gradual ID rat model. In addition, an inverse relationship between the P. carinii burden and the level of circulating CD4(+) or CD8(+) T lymphocytes was evidenced. This rat model may be used to study other opportunistic pathogens or even co-infections in a context of gradual ID.


Assuntos
Microbiologia do Ar , Modelos Animais de Doenças , Hospedeiro Imunocomprometido , Pneumocystis carinii/fisiologia , Pneumonia por Pneumocystis/microbiologia , Pneumonia por Pneumocystis/transmissão , Aerossóis , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Contagem de Colônia Microbiana , Dexametasona/administração & dosagem , Genes Fúngicos , Pulmão/microbiologia , Masculino , Pneumocystis carinii/efeitos dos fármacos , Pneumocystis carinii/crescimento & desenvolvimento , Pneumocystis carinii/isolamento & purificação , Ratos
6.
BMC Microbiol ; 14: 23, 2014 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-24495513

RESUMO

BACKGROUND: Histoplasma capsulatum and Pneumocystis organisms cause host infections primarily affecting the lung tissue. H. capsulatum is endemic in the United States of America and Latin American countries. In special environments, H. capsulatum is commonly associated with bat and bird droppings. Pneumocystis-host specificity has been primarily studied in laboratory animals, and its ability to be harboured by wild animals remains as an important issue for understanding the spread of this pathogen in nature. Bats infected with H. capsulatum or Pneumocystis spp. have been found, with this mammal serving as a probable reservoir and disperser; however, the co-infection of bats with both of these microorganisms has never been explored. To evaluate the impact of H. capsulatum and Pneumocystis spp. infections in this flying mammal, 21 bat lungs from Argentina (AR), 13 from French Guyana (FG), and 88 from Mexico (MX) were screened using nested-PCR of the fragments, employing the Hcp100 locus for H. capsulatum and the mtLSUrRNA and mtSSUrRNA loci for Pneumocystis organisms. RESULTS: Of the 122 bats studied, 98 revealed H. capsulatum infections in which 55 of these bats exhibited this infection alone. In addition, 51 bats revealed Pneumocystis spp. infection of which eight bats exhibited a Pneumocystis infection alone. A total of 43 bats (eight from AR, one from FG, and 34 from MX) were found co-infected with both fungi, representing a co-infection rate of 35.2% (95% CI = 26.8-43.6%). CONCLUSION: The data highlights the H. capsulatum and Pneumocystis spp.co-infection in bat population's suggesting interplay with this wild host.


Assuntos
Quirópteros , Coinfecção/veterinária , Histoplasma/isolamento & purificação , Histoplasmose/veterinária , Infecções por Pneumocystis/veterinária , Pneumocystis/isolamento & purificação , Animais , Argentina , Guiana , México , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Ribossômico/genética , Análise de Sequência de DNA
7.
Phytother Res ; 27(11): 1640-5, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23280633

RESUMO

Screening of the antifungal activities of ten Guadeloupean plants was undertaken to find new extracts and formulations against superficial mycoses such as onychomycosis, athlete's foot, Pityriasis versicolor, as well as the deep fungal infection Pneumocystis pneumonia. For the first time, the CMI of these plant extracts [cyclohexane, ethanol and ethanol/water (1:1, v/v)] was determined against five dermatophytes, five Candida species, Scytalidium dimidiatum, a Malassezia sp. strain and Pneumocystis carinii. Cytotoxicity tests of the most active extracts were also performed on an HaCat keratinocyte cell line. Results suggest that the extracts of Bursera simaruba, Cedrela odorata, Enterolobium cyclocarpum and Pluchea carolinensis have interesting activities and could be good candidates for developing antifungal formulations.


Assuntos
Antifúngicos/farmacologia , Arthrodermataceae/efeitos dos fármacos , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Asteraceae/química , Bursera/química , Candida/efeitos dos fármacos , Cedrela/química , Linhagem Celular , Fabaceae/química , Guadalupe , Humanos , Malassezia/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pneumocystis carinii/efeitos dos fármacos
8.
J Zoo Wildl Med ; 44(1): 15-20, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23505698

RESUMO

Histoplasma capsulatum is a dimorphic fungus that is widely distributed in the tropical or subtropical areas of the world and infects several mammalian hosts, mainly bats. Infective propagules grow in bat and bird droppings. A specific molecular marker, a highly sensitive fragment of a co-activator protein-coding gene (Hcp100), was used to detect H. capsulatum in lung samples of wild and captive bats from France using a nested polymerase chain reaction. To determine whether bats in France are potential carriers of H. capsulatum, 83 bats were sampled from two regions in France. Sixty-one specimens belonging to the Pteropus rodricensis (n = 45) and Rousettus aegyptiacus (n = 16) species were collected from a zoologic park (La Palmyre, western France). Twenty-two specimens were recovered from the Natural History Museum (Bourges) including the species Plecotus austriacus (n = 1), Pipistrellus pipistrellus (n = 3), and Nyctalus noctula (n = 18). From the lung DNA samples of 83 dead bats, only one sample of an N. noctula bat from Bourges amplified the H. capsulatum Hcp100 marker. The amplified product was sequenced and revealed a high similarity to the G217B H. capsulatum reference strain sequence that was deposited in the GenBank database. This finding suggests that H. capsulatum is an environmental pathogen in France that may infect bats.


Assuntos
Quirópteros/fisiologia , Histoplasma/isolamento & purificação , Histoplasmose/veterinária , Pneumopatias/microbiologia , Animais , Sequência de Bases , DNA Fúngico , França/epidemiologia , Histoplasmose/epidemiologia , Pneumopatias/epidemiologia , Reação em Cadeia da Polimerase/veterinária
9.
Appl Environ Microbiol ; 78(22): 8122-36, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23001662

RESUMO

Bats belong to a wide variety of species and occupy diversified habitats, from cities to the countryside. Their different diets (i.e., nectarivore, frugivore, insectivore, hematophage) lead Chiroptera to colonize a range of ecological niches. These flying mammals exert an undisputable impact on both ecosystems and circulation of pathogens that they harbor. Pneumocystis species are recognized as major opportunistic fungal pathogens which cause life-threatening pneumonia in severely immunocompromised or weakened mammals. Pneumocystis consists of a heterogeneous group of highly adapted host-specific fungal parasites that colonize a wide range of mammalian hosts. In the present study, 216 lungs of 19 bat species, sampled from diverse biotopes in the New and Old Worlds, were examined. Each bat species may be harboring a specific Pneumocystis species. We report 32.9% of Pneumocystis carriage in wild bats (41.9% in Microchiroptera). Ecological and behavioral factors (elevation, crowding, migration) seemed to influence the Pneumocystis carriage. This study suggests that Pneumocystis-host association may yield much information on Pneumocystis transmission, phylogeny, and biology in mammals. Moreover, the link between genetic variability of Pneumocystis isolated from populations of the same bat species and their geographic area could be exploited in terms of phylogeography.


Assuntos
Portador Sadio/veterinária , Variação Genética , Pulmão/microbiologia , Pneumocystis/classificação , Pneumocystis/genética , Pneumonia por Pneumocystis/veterinária , Animais , Portador Sadio/microbiologia , Quirópteros , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Mitocondrial/química , DNA Mitocondrial/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , Pneumocystis/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Análise de Sequência de DNA
10.
J Infect Dis ; 203(9): 1333-6, 2011 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-21372312

RESUMO

To better understand the diffusion of Pneumocystis in the environment, airborne shedding of Pneumocystis carinii in the surrounding air of experimentally infected rats was quantified by means of a real-time polymerase chain reaction assay, in parallel with the kinetics of P. carinii loads in their lungs. P. carinii DNA was detected in the air 1 week after infection and increased until 4-5 weeks after infection before stabilizing. A significant correlation was shown between lung burdens and the corresponding airborne levels, suggesting the possibility of estimating the fungal lung involvement through quantification of Pneumocystis in the exhaled air.


Assuntos
Microbiologia do Ar , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Animais , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , Modelos Animais de Doenças , Pulmão/microbiologia , Micologia/métodos , Reação em Cadeia da Polimerase/métodos , Ratos , Fatores de Tempo
11.
FEMS Yeast Res ; 11(1): 2-17, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21114625

RESUMO

At the end of the 20th century the unique taxonomically enigmatic entity called Pneumocystis carinii was identified as a heterogeneous group of microscopic Fungi, constituted of multiple stenoxenic biological entities largely spread across ecosystems, closely adapted to, and coevolving in parallel with, mammal species. The discoveries and reasoning that led to the current conceptions about the taxonomy of Pneumocystis at the species level are examined here. The present review also focuses on the biological, morphological and phylogenetical features of Pneumocystis jirovecii, Pneumocystis oryctolagi, Pneumocystis murina, P. carinii and Pneumocystis wakefieldiae, the five Pneumocystis species described until now, mainly on the basis of the phylogenetic species concept. Interestingly, Pneumocystis organisms exhibit a successful adaptation enabling them to dwell and replicate in the lungs of both immunocompromised and healthy mammals, which can act as infection reservoirs. The role of healthy carriers in aerial disease transmission is nowadays recognized as a major contribution to Pneumocystis circulation, and Pneumocystis infection of nonimmunosuppressed hosts has emerged as a public health issue. More studies need to be undertaken both on the clinical consequences of the presence of Pneumocystis in healthy carriers and on the intricate Pneumocystis life cycle to better define its epidemiology, to adapt existing therapies to each clinical context and to discover new drug targets.


Assuntos
Filogenia , Infecções por Pneumocystis/epidemiologia , Infecções por Pneumocystis/microbiologia , Pneumocystis/classificação , Animais , Humanos , Pulmão/microbiologia , Mamíferos , Pneumocystis/citologia , Pneumocystis/genética , Pneumocystis/patogenicidade , Infecções por Pneumocystis/patologia
12.
J Fungi (Basel) ; 7(11)2021 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-34829225

RESUMO

Histoplasmosis and pneumocystosis co-infections have been reported mainly in immunocompromised humans and in wild animals. The immunological response to each fungal infection has been described primarily using animal models; however, the host response to concomitant infection is unknown. The present work aimed to evaluate the pulmonary immunological response of patients with pneumonia caused either by Histoplasma capsulatum, Pneumocystis jirovecii, or their co-infection. We analyzed the pulmonary collectin and cytokine patterns of 131 bronchoalveolar lavage samples, which included HIV and non-HIV patients infected with H. capsulatum, P. jirovecii, or both fungi, as well as healthy volunteers and HIV patients without the studied fungal infections. Our results showed an increased production of the surfactant protein-A (SP-A) in non-HIV patients with H. capsulatum infection, contrasting with HIV patients (p < 0.05). Significant differences in median values of SP-A, IL-1ß, TNF-α, IFN-γ, IL-18, IL-17A, IL-33, IL-13, and CXCL8 were found among all the groups studied, suggesting that these cytokines play a role in the local inflammatory processes of histoplasmosis and pneumocystosis. Interestingly, non-HIV patients with co-infection and pneumocystosis alone showed lower levels of SP-A, IL-1ß, TNF-α, IFN-γ, IL-18, IL-17A, and IL-23 than histoplasmosis patients, suggesting an immunomodulatory ability of P. jirovecii over H. capsulatum response.

13.
J Nat Prod ; 73(7): 1313-7, 2010 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-20590148

RESUMO

Three compounds were isolated from Acnistus arborescens, a tree commonly used in South and Central America in traditional medicine against several infectious diseases, some of which are caused by fungi. Bioassay-guided fractionation of a MeOH extract of leaves, based on its anti-Pneumocystis carinii activity, led to the isolation of compounds 1-3. Mono- and bidimensional NMR analyses enabled identification of two new withanolides, (20R,22R)-5beta,6beta-epoxy-4beta,12beta,20-trihydroxy-1-oxowith-2-en-24-enolide (1) and (20R,22R)-16beta-acetoxy-3beta,4beta;5beta,6beta-diepoxy-12beta,20-dihydroxy-1-oxowith-24-enolide (2), and withanolide D (3). Antifungal activity on 13 fungi responsible for human infections (five dermatophytes, one nondermatophyte mold, six yeasts, and Pneumocystis carinii) was examined. Cytotoxicity of these compounds was also evaluated in vitro.


Assuntos
Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Plantas Medicinais/química , Vitanolídeos/isolamento & purificação , Vitanolídeos/farmacologia , Antifúngicos/química , Benzamidas , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Guadalupe , Humanos , Mesilato de Imatinib , Testes de Sensibilidade Microbiana , Estrutura Molecular , Piperazinas/farmacologia , Folhas de Planta/química , Pneumocystis carinii/efeitos dos fármacos , Pirimidinas/farmacologia , Solanaceae/química , Estereoisomerismo , Vitanolídeos/química
14.
J Eukaryot Microbiol ; 56(5): 446-53, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19737197

RESUMO

The separation of Pneumocystis carinii life-cycle stages while preserving infectivity is a hitherto unresolved challenge. We describe an original, reproducible, and efficient method for separating trophic from cystic forms of P. carinii using a high-speed cell sorter. The large amounts of highly purified (99.6+/-0.3%) infectious trophic and cystic forms can now be used to elucidate the poorly understood P. carinii life cycle.


Assuntos
Citometria de Fluxo/métodos , Pneumocystis carinii/classificação , Pneumocystis carinii/isolamento & purificação , Animais , Imunofluorescência/métodos , Humanos
15.
Mem Inst Oswaldo Cruz ; 104(3): 419-26, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19547866

RESUMO

First recognised as 'schizonts' of Trypanosoma cruzi, Pneumocystis organisms are now considered as part of an early-diverging lineage of Ascomycetes. As no robust long-term culture model is available, most data on the Pneumocystis cell cycle have stemmed from ultrastructural images of infected mammalian lungs. Although most fungi developing in animals do not complete a sexual cycle in vivo, Pneumocystis species constitute one of a few exceptions. Recently, the molecular identification of several key players in the fungal mating pathway has provided further evidence for the existence of conjugation and meiosis in Pneumocystisorganisms. Dynamic follow-up of stage-to-stage transition as well as studies of stage-specific proteins and/or genes would provide a better understanding of the still hypothetical Pneumocystislife cycle. Although difficult to achieve, stage purification seems a reasonable way forward in the absence of efficient culture systems. This mini-review provides a comprehensive overview of the historical milestones leading to the current knowledge available on the Pneumocystis life cycle.


Assuntos
Ciclo Celular/fisiologia , Genes Fúngicos Tipo Acasalamento/fisiologia , Estágios do Ciclo de Vida/fisiologia , Pneumocystis/crescimento & desenvolvimento , Animais , Ciclo Celular/genética , Genes Fúngicos Tipo Acasalamento/genética , Microscopia Eletrônica de Transmissão , Pneumocystis/genética , Pneumocystis/ultraestrutura
16.
JMIR Med Educ ; 5(1): e10766, 2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30973341

RESUMO

BACKGROUND: During their studies, pharmacy students must acquire the specific skills in clinical virology required for their subsequent professional practice. Recent experiments on teaching and learning in higher education have shown that hybrid courses strengthen the students' commitment to learning and enable high-quality knowledge acquisition. OBJECTIVE: This study concerned the design and deployment of a hybrid course that combines face-to-face and Web-based instruction in clinical virology for fourth-year pharmacy students. The study's objectives were to (1) measure the students' level of involvement in the course, (2) gauge their interest in this type of learning, and (3) highlight any associated difficulties. METHODS: The study included 194 fourth-year pharmacy students from the Lille Faculty of Pharmacy (University of Lille, Lille, France) between January and June 2017. The students followed a hybrid course comprising an online learning module and 5 tutorial sessions in which professional situations were simulated. The learning module and 3 online evaluation sessions were delivered via the Moodle learning management system. Each tutorial session ended with an evaluation. The number of Moodle log-ins, the number of views of learning resources, and the evaluation marks were recorded. The coefficient for the correlation between the marks in the online evaluation and those in the tutorials was calculated. The students' opinions and level of satisfaction were evaluated via a course questionnaire. RESULTS: The course's learning resources and Web pages were viewed 21,446 and 3413 times, respectively. Of the 194 students, 188 (96.9%) passed the course (ie, marks of at least 10 out of 20). There was a satisfactory correlation between the marks obtained in the online evaluations and those obtained after the tutorials. The course met the students' expectations in 53.2% of cases, and 57.4% of the students stated that they were able to work at their own pace. Finally, 26.6% of the students stated that they had difficulty organizing their work around this hybrid course. CONCLUSIONS: Our results showed that pharmacy students were strongly in favor of a hybrid course. The levels of attendance and participation were high. However, teachers must be aware that some students will encounter organizational difficulties.

17.
Int J Infect Dis ; 86: 65-72, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31207386

RESUMO

BACKGROUND: Histoplasma capsulatum and Pneumocystis jirovecii are respiratory fungal pathogens that principally cause pulmonary disease. Coinfection with both pathogens is scarcely reported. This study detected this coinfection using specific molecular methods for each fungus in the bronchoalveolar lavage (BAL) of patients from a tertiary care hospital. MATERIALS AND METHODS: BAL samples from 289 hospitalized patients were screened by PCR with specific markers for H. capsulatum (Hcp100) and P. jirovecii (mtLSUrRNA and mtSSUrRNA). The presence of these pathogens was confirmed by the generated sequences for each marker. The clinical and laboratory data for the patients were analyzed using statistical software. RESULTS: The PCR findings separated three groups of patients, where the first was represented by 60 (20.8%) histoplasmosis patients, the second by 45 (15.6%) patients with pneumocystosis, and the last group by 12 (4.2%) patients with coinfection. High similarity among the generated sequences of each species was demonstrated by BLASTn and neighbor-joining algorithms. The estimated prevalence of H. capsulatum and P. jirovecii coinfection was higher in HIV patients.


Assuntos
Coinfecção/epidemiologia , Histoplasmose/epidemiologia , Pneumocystis carinii , Pneumonia por Pneumocystis/epidemiologia , Adulto , Idoso , Lavagem Broncoalveolar , Feminino , Infecções por HIV/complicações , Histoplasma/genética , Histoplasma/isolamento & purificação , Histoplasmose/microbiologia , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Reação em Cadeia da Polimerase , Centros de Atenção Terciária
18.
Infect Genet Evol ; 8(5): 708-26, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18565802

RESUMO

The genus Pneumocystis comprises uncultured, highly diversified microfungal organisms able to attach specifically to type-I alveolar epithelial cells and to proliferate in pulmonary alveoli provoking severe pneumonitis. The pathogenic potential of Pneumocystis species, especially of the human-associated Pneumocystis jirovecii, has stimulated a growing interest in these peculiar microfungi. However, a comprehensive understanding of basic biology and pathogenic power of Pneumocystis organisms calls for their recognition as natural, complex entities, without reducing them to their pathogenic role. For many years, the entity named "Pneumocystis carinii" was considered like an anecdotal pulmonary pathogen able to cause pneumonia in immunosuppressed hosts. Only for the last years, marked genetic divergence was documented among the Pneumocystis strains of different mammals. Cross-infection experiments showed that Pneumocystis species are stenoxenous parasites. Mainly on the basis of the Phylogenetic Concept of Species, Pneumocystis strains were considered as genuine species. Five species were described: P. carinii and Pneumocystis wakefieldiae in rats, P. jirovecii in humans, Pneumocystis murina in mice, and Pneumocystis oryctolagi in rabbits. They also present distinctive phenotypic features. Molecular techniques have revealed a high prevalence of Pneumocystis colonization in wild mammals, probably resulting from active airborne horizontal and vertical (transplacental or aerial) transmission mechanisms. Cophylogeny is the evolutionary pattern for Pneumocystis species, which dwelt in the lungs of mammals for more than 100 million years. Consistently, Pneumocystis organisms exhibit successful adaptation to colonize the lungs of both immunocompromised and healthy hosts that can act as infection reservoir. Pneumocystis pneumonia, rarely reported in wild mammals, seems to be a rather unfrequent event. A larger spectrum of Pneumocystis infections related to the heterogeneous level of immune defence found in natural populations, is, however, expected. Pneumocystis infection of immunocompetent hosts emerges therefore as a relevant issue to human as well as animal health.


Assuntos
Evolução Biológica , Infecções por Pneumocystis/microbiologia , Pneumocystis/classificação , Pneumocystis/patogenicidade , Animais , Ecossistema , Interações Hospedeiro-Patógeno , Humanos , Imunocompetência , Estágios do Ciclo de Vida , Pulmão/microbiologia , Pneumocystis/genética , Pneumocystis/crescimento & desenvolvimento , Infecções por Pneumocystis/patologia , Infecções por Pneumocystis/transmissão , Coelhos , Especificidade da Espécie
19.
FEMS Microbiol Rev ; 30(6): 853-71, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17064284

RESUMO

The genus Pneumocystis comprises noncultivable, highly diversified fungal pathogens dwelling in the lungs of mammals. The genus includes numerous host-species-specific species that are able to induce severe pneumonitis, especially in severely immunocompromised hosts. Pneumocystis organisms attach specifically to type-1 epithelial alveolar cells, showing a high level of subtle and efficient adaptation to the alveolar microenvironment. Pneumocystis species show little difference at the light microscopy level but DNA sequences of Pneumocystis from humans, other primates, rodents, rabbits, insectivores and other mammals present a host-species-related marked divergence. Consistently, selective infectivity could be proven by cross-infection experiments. Furthermore, phylogeny among primate Pneumocystis species was correlated with the phylogeny of their hosts. This observation suggested that cophylogeny could explain both the current distribution of pathogens in their hosts and the speciation. Thus, molecular, ultrastructural and biological differences among organisms from different mammals strengthen the view of multiple species existing within the genus Pneumocystis. The following species were subsequently described: Pneumocystis jirovecii in humans, Pneumocystis carinii and Pneumocystis wakefieldiae in rats, and Pneumocystis murina in mice. The present work focuses on Pneumocystis oryctolagi sp. nov. from Old-World rabbits. This new species has been described on the basis of both biological and phylogenetic species concepts.


Assuntos
Pneumocystis/classificação , Pneumonia por Pneumocystis/veterinária , Animais , Animais Selvagens/microbiologia , França , Proteínas Fúngicas/genética , Genes Fúngicos , Pulmão/microbiologia , Microscopia Eletrônica , Dados de Sequência Molecular , Filogenia , Pneumocystis/genética , Pneumocystis/isolamento & purificação , Pneumocystis/patogenicidade , Pneumocystis/ultraestrutura , Pneumonia por Pneumocystis/microbiologia , Coelhos/microbiologia , Especificidade da Espécie
20.
Future Microbiol ; 13: 659-669, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29790796

RESUMO

AIM: To explore Aspergillus interactions with platelets in the blood, especially during clot formation. MATERIALS & METHODS: Aspergillus fumigatus resting or swollen conidia, germlings or hyphae were inoculated into blood sampled into tubes with or without anticoagulant. Interactions were explored using microscopy, and chemokine levels were determined. RESULTS: Anatomopathological examination of the clot revealed conidia and germlings colocalization with platelet aggregates, and neutrophil recruitment around aggregates. Transmission electron microscopy showed conidia and hyphae surrounded by neutrophils. Increased CCL5 and CXCL4 when conidia or germlings but not hyphae were added suggested they could be involved in neutrophil recruitment around aggregates. CONCLUSION: These data suggest platelets could trigger coagulopathy and activate neutrophils during aspergillosis. They open up new perspectives for aspergillosis management.


Assuntos
Aspergillus fumigatus/imunologia , Coagulação Sanguínea , Plaquetas/metabolismo , Neutrófilos/imunologia , Quimiocinas/análise , Voluntários Saudáveis , Humanos , Hifas/imunologia , Microscopia , Microscopia Eletrônica de Transmissão , Esporos Fúngicos/imunologia
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