22 research outputs found
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins
Background: TolT was originally described as a Trypanosoma cruzi molecule that accumulated on the trypomastigote flagellum bearing similarity to bacterial TolA colicins receptors. Preliminary biochemical studies indicated that TolT resolved in SDS-PAGE as ~3–5 different bands with sizes between 34 and 45 kDa, and that this heterogeneity could be ascribed to differences in polypeptide glycosylation. However, the recurrent identification of TolT-deduced peptides, and variations thereof, in trypomastigote proteomic surveys suggested an intrinsic TolT complexity, and prompted us to undertake a thorough reassessment of this antigen. Methods/Principle findings: Genome mining exercises showed that TolT constitutes a larger-than-expected family of genes, with at least 12 polymorphic members in the T. cruzi CL Brener reference strain and homologs in different trypanosomes. According to structural features, TolT deduced proteins could be split into three robust groups, termed TolT-A, TolT-B, and TolT-C, all of them showing marginal sequence similarity to bacterial TolA proteins and canonical signatures of surface localization/membrane association, most of which were herein experimentally validated. Further biochemical and microscopy-based characterizations indicated that this grouping may have a functional correlate, as TolT-A, TolT-B and TolT-C molecules showed differences in their expression profile, sub-cellular distribution, post-translational modification(s) and antigenic structure. We finally used a recently developed fluorescence magnetic beads immunoassay to validate a recombinant protein spanning the central and mature region of a TolT-B deduced molecule for Chagas disease serodiagnosis. Conclusion/Significance: This study unveiled an unexpected genetic and biochemical complexity within the TolT family, which could be exploited for the development of novel T. cruzi biomarkers with diagnostic/therapeutic applications.Fil: Lobo, Mabel Maite. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Balouz, Virginia. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Melli, Luciano Jorge. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Carlevaro, Giannina Alejandra. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Cortina, MarÃa Eugenia. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Camara, MarÃa de los Milagros. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Canepa, Gaspar Exequiel. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Carmona, Santiago Javier. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Altcheh, Jaime Marcelo. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; Argentina. Consejo Nacional de Investigaciones CientÃficas y Técnicas; ArgentinaFil: Campetella, Oscar Eduardo. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Ciocchini, Andres Eduardo. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Agüero, Fernan Gonzalo. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Mucci, Juan Sebastián. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Buscaglia, Carlos Andres. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San MartÃn. Instituto de Investigaciones Biotecnológicas; Argentin
The Differential Interaction of Brucella and Ochrobactrum with Innate Immunity Reveals Traits Related to the Evolution of Stealthy Pathogens
International audienceBACKGROUND: During evolution, innate immunity has been tuned to recognize pathogen-associated molecular patterns. However, some alpha-Proteobacteria are stealthy intracellular pathogens not readily detected by this system. Brucella members follow this strategy and are highly virulent, but other Brucellaceae like Ochrobactrum are rhizosphere inhabitants and only opportunistic pathogens. To gain insight into the emergence of the stealthy strategy, we compared these two phylogenetically close but biologically divergent bacteria. METHODOLOGY/PRINCIPAL FINDINGS: In contrast to Brucella abortus, Ochrobactrum anthropi did not replicate within professional and non-professional phagocytes and, whereas neutrophils had a limited action on B. abortus, they were essential to control O. anthropi infections. O. anthropi triggered proinflammatory responses markedly lower than Salmonella enterica but higher than B. abortus. In macrophages and dendritic cells, the corresponding lipopolysaccharides reproduced these grades of activation, and binding of O. anthropi lipopolysaccharide to the TLR4 co-receptor MD-2 and NF-kappaB induction laid between those of B. abortus and enteric bacteria lipopolysaccharides. These differences correlate with reported variations in lipopolysaccharide core sugars, sensitivity to bactericidal peptides and outer membrane permeability. CONCLUSIONS/SIGNIFICANCE: The results suggest that Brucellaceae ancestors carried molecules not readily recognized by innate immunity, so that non-drastic variations led to the emergence of stealthy intracellular parasites. They also suggest that some critical envelope properties, like selective permeability, are profoundly altered upon modification of pathogen-associated molecular patterns, and that this represents a further adaptation to the host. It is proposed that this adaptive trend is relevant in other intracellular alpha-Proteobacteria like Bartonella, Rickettsia, Anaplasma, Ehrlichia and Wolbachia
Proposta metodológica para exame, registo e representação gráfica dos brocados aplicados
La falta de una fórmula común entre los profesionales y estudiosos que trabajan con los brocados aplicados ha llevado al grupo de trabajo europeo de especialistas en escultura policromada a definir un procedimiento para el examen, registro y representación gráfica que permita profundizar en el estudio de esta delicada y frágil técnica a través de una información normalizada. Este trabajo recoge los aspectos del brocado aplicado que han de ser examinados; cómo éstos pueden ser recogidos y registrados atendiendo a las condiciones y estrategias más inocuas para la obra y propone, por último, un procedimiento para su representación gráfica.The lack of a common formula among professionals and scholars for the study of applied brocades has led the European working group of specialists in polychrome sculpture to define a standardized procedure for examination, registration and graphic representation that allows a deeper understanding of this delicate and fragile technique. This work gathers the aspects of the applied brocade that have to be examined; how these can be collected and registered according to the most harmless conditions and strategies and proposes a procedure for their graphic representation.A falta de uma fórmula comum entre os profissionais e estudiosos que trabalham com brocados aplicados levou o grupo de trabalho europeu de especialistas em escultura policromada a definir um procedimento de exame, registo e representação gráfica que permita aprofundar o estudo desta delicada e frágil técnica através de uma informação normalizada. Este trabalho reúne os aspetos do brocado aplicado que devem ser examinados; como estes podem ser recolhidos e registados de acordo com as condições e estratégias mais inócuas para a obra e propõe, por fim, um procedimento para a sua representação gráfica
A New Multidisciplinary Home Care Telemedicine System to Monitor Stable Chronic Human Immunodeficiency Virus-Infected Patients: A Randomized Study
BACKGROUND:
Antiretroviral therapy has changed the natural history of human immunodeficiency virus (HIV) infection in developed countries, where it has become a chronic disease. This clinical scenario requires a new approach to simplify follow-up appointments and facilitate access to healthcare professionals.
METHODOLOGY:
We developed a new internet-based home care model covering the entire management of chronic HIV-infected patients. This was called Virtual Hospital. We report the results of a prospective randomised study performed over two years, comparing standard care received by HIV-infected patients with Virtual Hospital care. HIV-infected patients with access to a computer and broadband were randomised to be monitored either through Virtual Hospital (Arm I) or through standard care at the day hospital (Arm II). After one year of follow up, patients switched their care to the other arm. Virtual Hospital offered four main services: Virtual Consultations, Telepharmacy, Virtual Library and Virtual Community. A technical and clinical evaluation of Virtual Hospital was carried out.
FINDINGS:
Of the 83 randomised patients, 42 were monitored during the first year through Virtual Hospital (Arm I) and 41 through standard care (Arm II). Baseline characteristics of patients were similar in the two arms. The level of technical satisfaction with the virtual system was high: 85% of patients considered that Virtual Hospital improved their access to clinical data and they felt comfortable with the videoconference system. Neither clinical parameters [level of CD4+ T lymphocytes, proportion of patients with an undetectable level of viral load (p = 0.21) and compliance levels >90% (p = 0.58)] nor the evaluation of quality of life or psychological questionnaires changed significantly between the two types of care.
CONCLUSIONS:
Virtual Hospital is a feasible and safe tool for the multidisciplinary home care of chronic HIV patients. Telemedicine should be considered as an appropriate support service for the management of chronic HIV infection.
TRIAL REGISTRATION:
Clinical-Trials.gov: NCT01117675
Acute toxicity of zinc and arsenic to the warmwater aquatic oligochaete Branchiura sowerbyi as compared to its coldwater counterpart Tubifex tubifex (Annelida, Clitellata)
Purpose This study aimed at evaluating the acute effects of arsenic and zinc to the warmwater aquatic oligochaete Branchiura sowerbyi. Relative sensitivity with the coldwater species Tubifex tubifex was compared. Implications for the use of B. sowerbyi in the risk assessment of sediments in the tropics are discussed.
Materials and methods Water-only (96 h) and sediment (14 days) toxicity tests were conducted with both species evaluating a concentration series of arsenic and zinc. The tests were conducted considering the environmental conditions in the natural habitat of T. tubifex (predominantly temperate) and B. sowerbyi (predominantly tropical). Both lethal and sublethal endpoints (autotomy of the posterior body parts, abnormal behavior and appearance) were determined in the tests. The lethal (LC10 and LC50) and effect (EC10 and EC50) concentrations were also determined to assess metal sensitivity for both species.
Results and discussion Both test species were more sensitive to Zn than As in water-only tests, which is in agreement with previous studies evaluating the toxicity of these metals to aquatic oligochaetes. Sublethal effects were generally noted at concentrations lower than those leading to mortality. The warmwater oligochaete B. sowerbyi was more sensitive to both metals tested than the coldwater species T. tubifex.
Conclusions Study findings support the need for using indigenous tropical species in risk assessments in the tropics. In addition, sublethal effect parameters should be included in toxicity testing with aquatic oligochaetes.We would like to thank the National Council for Scientific and Technological Development (CPNq) and the Coordination of Higher Education
Personnel Improvement (CAPES) from the Brazilian Government for the scholarships
granted (CNPq: 140771/2010-7; CAPES: PDSE 9805/11-7). This work was possible
thanks to the support from the Education and Science Ministry research project (MEC
CGL2008-04502/BOS) and from the Basque Government (IT-405-10)...Dr. L. Méndez-Fernández was
supported by a postdoctoral fellowship from the University of the Basque Country
Bioaccumulation and chronic toxicity of arsenic and zinc in the aquatic oligochaetes Branchiura sowerbyi and Tubifex tubifex (Annelida, Clitellata)
Oligochaetes feed on bulk sediment and penetrate the sediment through the construction of burrows, making them especially vulnerable to sediment metal contamination. However, the few oligochaete species that have been tested to date are almost exclusively temperate test species. Although the warmwater adapted species Branchiura sowerbyi has been indicated as a promising candidate for tropical sediment toxicity testing, few (especially chronic) studies have been conducted so far to confirm this. Therefore, the aim of the present study was to evaluate the bioaccumulation and chronic 28d lethal and sublethal toxicity of arsenic (As) and zinc (Zn) to both the warmwater-adapted B. sowerbyi and the coldwater-adapted oligochaete Tubifex tubifex for comparison. Arsenic was more toxic to both oligochaete species than Zn. Inter- and intra-species variability in toxicity values of the two test species and other benthic invertebrates was within an order of magnitude. However, B. sowerbyi was the most sensitive species to As even for sediment concentration (EC50: 36.6 ± 2.1 μg/g and 147.1 ± 21.7 μg/g, for B. sowerbyi and T. tubifex, respectively) and for tissue concentration (ER50: 9.2 ± 0.9 μg/g and 887.0 ± 35.0 μg/g, for B. sowerbyi and T. tubifex, respectively). Finally, the Tissue Residue-effects Approach (TRA) using Effective Tissue Residues appears to be a promising way forward in advancing in this since it considers internal body concentrations.We would like to thank the National Council for Scientific and Technological Development (CPNq) and the Coordination of Higher Education Personnel Improvement (CAPES) from Brazilian Government for the scholarships granted (CNPq: 140771/2010-7; CAPES: PDSE 9805/11-7). This work was possible thanks to the support from the Education and Science Ministry research project (MEC CGL2008-04502/BOS) and from the Basque Government (IT-405-10). Financial support was also provided to M. Daam by the Portuguese government (Fundação para a Cien̂cia e Tecnologia; FCT) through the research unit UIDB/04085/2020 (CENSE)
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins.
BACKGROUND:TolT was originally described as a Trypanosoma cruzi molecule that accumulated on the trypomastigote flagellum bearing similarity to bacterial TolA colicins receptors. Preliminary biochemical studies indicated that TolT resolved in SDS-PAGE as ~3-5 different bands with sizes between 34 and 45 kDa, and that this heterogeneity could be ascribed to differences in polypeptide glycosylation. However, the recurrent identification of TolT-deduced peptides, and variations thereof, in trypomastigote proteomic surveys suggested an intrinsic TolT complexity, and prompted us to undertake a thorough reassessment of this antigen. METHODS/PRINCIPLE FINDINGS:Genome mining exercises showed that TolT constitutes a larger-than-expected family of genes, with at least 12 polymorphic members in the T. cruzi CL Brener reference strain and homologs in different trypanosomes. According to structural features, TolT deduced proteins could be split into three robust groups, termed TolT-A, TolT-B, and TolT-C, all of them showing marginal sequence similarity to bacterial TolA proteins and canonical signatures of surface localization/membrane association, most of which were herein experimentally validated. Further biochemical and microscopy-based characterizations indicated that this grouping may have a functional correlate, as TolT-A, TolT-B and TolT-C molecules showed differences in their expression profile, sub-cellular distribution, post-translational modification(s) and antigenic structure. We finally used a recently developed fluorescence magnetic beads immunoassay to validate a recombinant protein spanning the central and mature region of a TolT-B deduced molecule for Chagas disease serodiagnosis. CONCLUSION/SIGNIFICANCE:This study unveiled an unexpected genetic and biochemical complexity within the TolT family, which could be exploited for the development of novel T. cruzi biomarkers with diagnostic/therapeutic applications
Trypanosoma cruzi surface mucins are involved in the attachment to the Triatoma infestans rectal ampoule.
BackgroundTrypanosoma cruzi, the agent of Chagas disease, is a protozoan parasite transmitted to humans by blood-sucking triatomine vectors. However, and despite its utmost biological and epidemiological relevance, T. cruzi development inside the digestive tract of the insect remains a poorly understood process.Methods/principle findingsHere we showed that Gp35/50 kDa mucins, the major surface glycoproteins from T. cruzi insect-dwelling forms, are involved in parasite attachment to the internal cuticle of the triatomine rectal ampoule, a critical step leading to its differentiation into mammal-infective forms. Experimental evidence supporting this conclusion could be summarized as follows: i) native and recombinant Gp35/50 kDa mucins directly interacted with hindgut tissues from Triatoma infestans, as assessed by indirect immunofluorescence assays; ii) transgenic epimastigotes over-expressing Gp35/50 kDa mucins on their surface coat exhibited improved attachment rates (~2-3 fold) to such tissues as compared to appropriate transgenic controls and/or wild-type counterparts; and iii) certain chemically synthesized compounds derived from Gp35/50 kDa mucins were able to specifically interfere with epimastigote attachment to the inner lining of T. infestans rectal ampoules in ex vivo binding assays, most likely by competing with or directly blocking insect receptor(s). A solvent-exposed peptide (smugS peptide) from the Gp35/50 kDa mucins protein scaffolds and a branched, Galf-containing trisaccharide (Galfβ1-4[Galpβ1-6]GlcNAcα) from their O-linked glycans were identified as main adhesion determinants for these molecules. Interestingly, exogenous addition of a synthetic Galfβ1-4[Galpβ1-6]GlcNAcα derivative or of oligosaccharides containing this structure impaired the attachment of Dm28c but not of CL Brener epimastigotes to triatomine hindgut tissues; which correlates with the presence of Galf residues on the Gp35/50 kDa mucins' O-glycans on the former but not the latter parasite clone.Conclusion/significanceThese results provide novel insights into the mechanisms underlying T. cruzi-triatomine interplay, and indicate that inter-strain variations in the O-glycosylation of Gp35/50 kDa mucins may lead to differences in parasite differentiation and hence, in parasite transmissibility to the mammalian host. Most importantly, our findings point to Gp35/50 kDa mucins and/or the Galf biosynthetic pathway, which is absent in mammals and insects, as appealing targets for the development of T. cruzi transmission-blocking strategies