Molecular diagnosis and typing of Trypanosoma cruzi populations and lineages in cerebral Chagas disease in a patient with AIDS

Trypanosoma cruzi DNA was amplified from an intracranial biopsy and peripheral blood of an HIV patient with encephalitis; this episode was indicative of AIDS and congenital Chagas disease. The analysis of a microsatellite locus revealed a multiclonal parasite population at the brain lesion with a more complex minicircle signature than that profiled in blood using restriction fragment length polymorphism (RFLP)-PCR and low stringency single primer (LSSP) PCR. Interestingly, different sublineages of T. cruzi II were detected in blood and brain by means of spliced-leader and 24s ribosomal-DNA amplifications. Quantitative-competitive PCR monitored the decrease of parasitic load during treatment and secondary prophylaxis with benznidazole. The synergy between parasiticidal plus antiretroviral treatments probably allowed the patient a longer survival than usually achieved in similar episodes. This is the first case report demonstrating a differential distribution of natural parasite populations and sublineages in Chagas disease reactivation, showing the proliferation of cerebral variants not detectable in peripheral blood.Fil: Burgos, Juan Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Bergher, Sandra B.. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Freitas, Jorge M.. Universidade Federal de Minas Gerais; BrasilFil: Bisio, Margarita María Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Altcheh, Jaime Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; ArgentinaFil: Teijeiro, Ricardo. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Begher, Sandra B.. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Freilij, Hector León. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; ArgentinaFil: Deccarlini, Florencia. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Levalle, Jorge. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Lopez Alcoba, Horacio. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Burgos, Juan Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Levin, Mariano Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Duffy, Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Macedo, Andrea M.. Universidade Federal de Minas Gerais; BrasilFil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentin

New textile for personal protective equipment—plasma chitosan/silver nanoparticles nylon fabric

Fabric structures are prone to contamination with microorganisms, as their morphology and ability to retain moisture creates a proper environment for their growth. In this work, a novel, easily processed and cheap coating for a nylon fabric with antimicrobial characteristics was devel- oped. After plasma treatment, made to render the fabric surface more reactive sites, the fabric was impregnated with chitosan and silver nanoparticles by simply dipping it into a mixture of different concentrations of both components. Silver nanoparticles were previously synthesized using the Lee–Meisel method, and their successful obtention was proven by UV–Vis, showing the presence of the surface plasmon resonance band at 410 nm. Nanoparticles with 25 nm average diameter observed by STEM were stable, mainly in the presence of chitosan, which acted as a surfactant for silver nanoparticles, avoiding their aggregation. The impregnated fabric possessed bactericidal activ- ity higher for Gram-positive Staphylococcus aureus than for Gram-negative Pseudomonas aeruginosa bacteria for all combinations. The percentage of live S. aureus and P. aeruginosa CFU was reduced to less than 20% and 60%, respectively, when exposed to each of the coating combinations. The effect was more pronounced when both chitosan and silver were present in the coating, suggesting an effective synergy between these components. After a washing process, the antimicrobial effect was highly reduced, suggesting that the coating is unstable after washing, being almost completely removed from the fabric. Nevertheless, the new-coated fabric can be successfully used in single-use face masks. To our knowledge, the coating of nylon fabrics intended for face-mask material with both agents has never been reported.This study was supported by the Portuguese Foundation for Science and Technology (FCT), under the scope of the strategic funding of UIDB/04469/2020 unit, and BioTecNorte operation (NORTE-01-0145-FEDER-000004). funded by the European Regional Development, Fund under the scope of Norte2020—Programa Operacional Regional do Norte

Trypanosoma cruzi benznidazole susceptibility in vitro does not predict the therapeutic outcome of human Chagas disease

Therapeutic failure of benznidazole (BZ) is widely documented in Chagas disease and has been primarily associated with variations in the drug susceptibility of Trypanosoma cruzi strains. In humans, therapeutic success has been assessed by the negativation of anti-T. cruzi antibodies, a process that may take up to 10 years. A protocol for early screening of the drug resistance of infective strains would be valuable for orienting physicians towards alternative therapies, with a combination of existing drugs or new anti-T. cruzi agents. We developed a procedure that couples the isolation of parasites by haemoculture with quantification of BZ susceptibility in the resultant epimastigote forms. BZ activity was standardized with reference strains, which showed IC50 to BZ between 7.6-32 µM. The assay was then applied to isolates from seven chronic patients prior to administration of BZ therapy. The IC50 of the strains varied from 15.6 ± 3-51.4 ± 1 µM. Comparison of BZ susceptibility of the pre-treatment isolates of patients considered cured by several criteria and of non-cured patients indicates that the assay does not predict therapeutic outcome. A two-fold increase in BZ resistance in the post-treatment isolates of two patients was verified. Based on the profile of nine microsatellite loci, sub-population selection in non-cured patients was ruled out.FAPESPCNP

Empowering minority women: Autonomy versus participation

Feminist attempts to empower women within their own cultural traditions have employed two broad strategies: authentic choice and participation. This paper argues that the methodological problems that beset the authentic choice strategy tell in favour of the participation approach. However, proponents of the participation strategy have failed to pay sufficient attention to the background conditions that need to be met if women are to make effective use of the institutional mechanisms their models advocate. If women are to be effective political agents at least some of the most serious structural inequalities that women face must be addressed. A nuanced statement of the participation strategy must therefore take account of long-standing feminist concerns regarding economic equality and access to resources. While this approach falls short of the demanding conditions for democratic citizenship implicit in the authentic choice strategy, it none the less places significant limits on the scope of participatory strategies and links the goal of empowering women within their own cultural traditions to wider feminist struggles to secure greater economic equality for women in general

Comfort and infection control of chitosan-impregnated cotton gauze as wound dressing

The aim of this study was to evaluate the thermo-physiological comfort properties of surgical cotton gauze coated with chitosan (CH) and its effectiveness for the prevention of bacterial colonization. Gauze was coated with CH at mass fractions of 0.50, 0.25, 0.125, 0.10, 0.063 wt% and the friction, flexibility, thermal, moisture management and mechanical properties were evaluated. The best performing gauze in terms of comfort (0.125 wt%) was further evaluated for its ability to inhibit the growth of microorganisms such as bacteria and yeast. Results indicate that the functionalized medical gauze could induce low friction on the wound bed allowing a good degree of moisture and high absorption capacity of wound exudates. Moreover, it shows antimicrobial properties against medical-relevant pathogens. This biofunctional medical gauze demonstrates to deliver an efficient antimicrobial coating and promote the best conditions for maintenance of the wound microenvironment.Z acknowledges funding from FCT - Fundação para a Ciência e a Tecnologia within the scope of the project POCI- 01-0145-FEDER-007136 and UID/CTM/00264. A. Zille also acknowledges financial support of the FCT through an Investigator FCT Research contract (IF/00071/2015). JS acknowledge CAPES Foundation, Ministry of Education of Brazil, Proc. no 8976/13-9 and the Department of Textile Engineering of the University of Minho, Portugal. The work regarding the biological analysis was supported by the Programa Operacional, Fatores de competitividade - COMPETE and by national funds through FCT on the scope of the projects PTDC/SAU-MIC/119069/2010, RECI/EBB-EBI/ 0179/2012 and PEst-OE/EQB/LA0023/2013. PT acknowledges SFRH/BPD/86732/2012 grant. The authors thank the Project BioHealth - Biotechnology and Bioengineering approaches to improve health quality, Ref. NORTE-07-0124-FEDER- 000027, co-funded by the Programa Operacional Regional do Norte (ON.2 - O Novo Norte), QREN, FEDER.info:eu-repo/semantics/publishedVersio

The MHC Gene Region of Murine Hosts Influences the Differential Tissue Tropism of Infecting Trypanosoma cruzi Strains

We have previously demonstrated that both parasite genetic variability and host genetic background were important in determining the differential tissue distribution of the Col1.7G2 and JG T. cruzi monoclonal strains after artificial infections in mice. We observed that the JG strain was most prevalent in hearts of mouse lineages with the MHC haplotype H-2d (BALB/c and DBA2), while Col1.7G2 was predominant in hearts from C57BL/6 mice, which have the H-2b haplotype. To assess whether the MHC gene region indeed influenced tissue tropism of T. cruzi, we used the same two parasite strains to infect C57BL/6 (H-2b) and C57BLKS/J (H-2d) mice; the latter strain results from the introgression of DBA2 MHC region into the C57BL/6 background. We also performed ex vivo infections of cardiac explants from four congenic mice lineages with the H-2b and H-2d haplotypes arranged in two different genetic backgrounds: C57BLKS/J (H-2d) versus C57BL/6 (H-2b) and BALB/c (H-2d) versus BALB/B10-H2b (H-2b). In agreement with our former observations, Col1.7G2 was predominant in hearts from C57BL/6 mice (H-2b), but we observed a clear predominance of the JG strain in hearts from C57BLKS/J animals (H-2d). In the ex vivo experiments Col1.7G2 also prevailed in explants from H-2b animals while no predominance of any of the strains was observed in H-2d mice explants, regardless of the genetic background. These observations clearly demonstrate that the MHC region influences the differential tissue distribution pattern of infecting T. cruzi strains, which by its turn may be in a human infection the determinant for the clinical forms of the Chagas disease

The kSORT assay to detect renal transplant patients at high risk for acute rejection: results of the multicenter AART study

Abstract Background: Development of noninvasive molecular assays to improve disease diagnosis and patient monitoring is a critical need. In renal transplantation, acute rejection (AR) increases the risk for chronic graft injury and failure. Noninvasive diagnostic assays to improve current late and nonspecific diagnosis of rejection are needed. We sought to develop a test using a simple blood gene expression assay to detect patients at high risk for AR. Methods and Findings: We developed a novel correlation-based algorithm by step-wise analysis of gene expression data in 558 blood samples from 436 renal transplant patients collected across eight transplant centers in the US, Mexico, and Spain between 5 February 2005 and 15 December 2012 in the Assessment of Acute Rejection in Renal Transplantation (AART)study. Gene expression was assessed by quantitative real-time PCR (QPCR) in one center. A 17-gene set the Kidney Solid Organ Response Test (kSORT) was selected in 143 samples for AR classification using discriminant analysis (area under the receiver operating characteristic curve [AUC] = 0.94; 95% CI 0.91-0.98), validated in 124 independent samples (AUC = 0.95; 95% CI 0.88-1.0) and evaluated for AR prediction in 191 serial samples, where it predicted AR up to 3 mo prior to detection by the current gold standard (biopsy). A novel reference-based algorithm (using 13 12-gene models) was developed in 100 independent samples to provide a numerical AR risk score, to classify patients as high risk versus low risk for AR. kSORT was able to detect AR in blood independent of age, time post-transplantation, and sample source without additional data normalization; AUC = 0.93 (95% CI 0.86-0.99). Further validation of kSORT is planned in prospective clinical observational and interventional trials. Conclusions: The kSORT blood QPCR assay is a noninvasive tool to detect high risk of AR of renal transplants

Size effects on antimicrobial efficiency of DBD plasma coated silver nanoparticles on textiles

This work studies the surface characteristics, the antimicrobial activity and the aging effect, of plasma pre-treated polyamide 6,6 fabrics (PA66) coated with silver nanoparticles (AgNPs), with the aim to identify the optimum size of nanosilver exhibiting antibacterial properties suitable for manufacturing of hospital textiles. The release of bactericidal Ag+ ions from the 10, 20, 40, 60 and 100 nm AgNPs-coated PA66 surface were function of the particles size, number and aging. Plasma pre-treatment promoted both ionic and covalent interactions between AgNPs and the formed oxygen species on the fibers (Figure 1), favoring the deposition of smaller in diameter AgNPs that consequently showed better immediate and durable antimicrobial effect against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria. Surprisingly, after 30 days of aging, a comparable bacterial growth inhibition was achieved for all the fibers treated with AgNPs of <100 nm in size. The Ag+ in the coatings also favored the electrostatic stabilization of the plasma-induced functional groups on the PA66 surface, thereby retarding the aging process (Figure 2). At the same time, the size-related ratio Ag+/Ag0 of the AgNPs between 40 and 60 nm allowed for controlled release of Ag+ rather than bulk silver. Overall, the results suggest that instead of reducing the AgNPs size, which is associated to higher toxicity, similar long-term effects can be achieved with larger NPs (40-60 nm), even in lower concentrations. Since the antimicrobial efficiency of AgNPs larger than 30 nm is mainly ruled by the release of Ag+ over time and not by the size and number of the AgNPs, this parameter is crucial for the development of efficient antimicrobial coatings on plasma-treated surfaces, and contribution to the safety and durability of clothing used in clinical settings

Size and aging effects on antimicrobial efficiency of silver nanoparticles coated on polyamide fabrics activated by atmospheric DBD plasma

Recently, renewed interest has arisen in silver nanopar@cles for biomedical devices because of their high surface energy, enhanced physicochemical and biological proper@es and extremely large surface area, which provides beAer contact with microorganisms. Atmospheric plasma is an alterna@ve and cost- compe@@ve method to wet chemical nanopar@cles deposi@on methods, avoiding the need of toxic solvents, expensive vacuum equipment and allowing con@nuous and uniform processing of material surfaces. However, there are no reports on the size and @me-dependent an@microbial, physical and chemical surface effects of the silver nanopar@cles immobilized on plasma func@onalized polymers. Thus, the purposes of this study were: (i) the silver nanopar@cle size and aging effects aCer 30 days on the an@microbial ac@vity aCer deposi@on onto DBD plasma-treated polyamide 6,6 fabrics, and (ii) the aging effect on the physico-chemical binding mechanism between different sized silver nanopar@cles and the plasma treaded polyamide 6,6. Five different in size commercial silver nanopar@cles have been employed (10, 20, 40 60 and 100 nm).This work was funded by Portuguese Founda@on for Science and Technology FCT/MCTES (PIDDAC) and co-financed by European funds (FEDER) through the PT2020 program, research project M-ERA-NET/0006/2014 and COMPETE program through FCT within the scope of the project POCI-01-0145-FEDER-007136 and UID/CTM/00264.info:eu-repo/semantics/publishedVersio