31 results on '"Nogueira CL"'
Search Results
2. Tissue plasminogen activator-treated patients with acute ischemic stroke in the pioneer public service of Rio de Janeiro: a comparative profile with the NINDS study
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Missaka, H, Almeida, JE, Figueiredo, PC, Nogueira, CL, Julião, VR, Alencar, JLL, Lannes, RS, Fernandes, SL, Caetani, G, Abrantes, J, Bevilacqua, TD, Antonucci, V, Pinto, PHCF, and Divan-Filho, S
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- 2011
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3. Conhecimentos sobre n�meros irracionais mobilizados por alunos brasileiros e franceses�: um estudo com alunos de 9� ano e troisi�me
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Rezende, Veridiana, primary, Ignatius Nogueira, Cl�lia Maria, additional, and Magalh�es de Freitas, Jos� Luiz, additional
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- 2015
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4. A sociologia da educação de Pierre Bourdieu: limites e contribuições
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Nogueira Cláudio Marques Martins and Nogueira Maria Alice
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Sociologia da Educação ,Bourdieu ,Família. Escola ,Education (General) ,L7-991 - Abstract
O artigo destaca as contribuições e aponta alguns limites da Sociologia da Educação de Pierre Bourdieu. Na primeira parte, são analisadas as reflexões do autor sobre a relação entre herança familiar (sobretudo, cultural) e desempenho escolar. Na segunda parte, são discutidas suas teses sobre o papel da escola na reprodução e legitimação das desigualdades sociais.
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- 2002
5. Processo de obtenção de pré-impregnados poliméricos termoplásticos via moldagem por compressão a quente
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Nogueira Clara L., Marlet José M. F., and Rezende Mirabel C.
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Compósitos termoplásticos ,pré-impregnados ,moldagem por compressão a quente ,poli(éter-éter-cetona) ,poli(éter-imida) ,poli(sulfeto de fenileno) ,poli(sulfona) ,Chemical technology ,TP1-1185 - Abstract
Os compósitos poliméricos à base de termoplásticos avançados são usados em aplicações estruturais devido à uma combinação favorável de baixa massa específica e desempenho mecânico elevado. Uma das rotas para o processamento desses compósitos é via laminação e consolidação adequadas de pré-impregnados poliméricos termoplásticos. Este trabalho apresenta o estudo da obtenção de pré-impregnados termoplásticos via moldagem por compressão a quente, mostrando os parâmetros temperatura e pressão utilizados no processamento. Como matrizes poliméricas foram utilizadas a poli(éter-éter-cetona), a poli(éter-imida), o poli(sulfeto de fenileno) e a poli(sulfona) e como material de reforço tecido de fibras de carbono.
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- 1999
6. Treatment of recurrent digital scar contracture in paediatric patients by proximal phalangeal island flap
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Bertelli, J. and Nogueira, Cl.
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- 1997
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7. Potential of bacteriophage proteins as recognition molecules for pathogen detection.
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Costa SP, Nogueira CL, Cunha AP, Lisac A, and Carvalho CM
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- Proteins metabolism, Bacteriophages
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Bacterial pathogens are leading causes of infections with high mortality worldwide having a great impact on healthcare systems and the food industry. Gold standard methods for bacterial detection mainly rely on culture-based technologies and biochemical tests which are laborious and time-consuming. Regardless of several developments in existing methods, the goal of achieving high sensitivity and specificity, as well as a low detection limit, remains unaccomplished. In past years, various biorecognition elements, such as antibodies, enzymes, aptamers, or nucleic acids, have been widely used, being crucial for the pathogens detection in different complex matrices. However, these molecules are usually associated with high detection limits, demand laborious and costly production, and usually present cross-reactivity. (Bacterio)phage-encoded proteins, especially the receptor binding proteins (RBPs) and cell-wall binding domains (CBDs) of endolysins, are responsible for the phage binding to the bacterial surface receptors in different stages of the phage lytic cycle. Due to their remarkable properties, such as high specificity, sensitivity, stability, and ability to be easily engineered, they are appointed as excellent candidates to replace conventional recognition molecules, thereby contributing to the improvement of the detection methods. Moreover, they offer several possibilities of application in a variety of detection systems, such as magnetic, optical, and electrochemical. Herein we provide a review of phage-derived bacterial binding proteins, namely the RBPs and CBDs, with the prospect to be employed as recognition elements for bacteria. Moreover, we summarize and discuss the various existing methods based on these proteins for the detection of nosocomial and foodborne pathogens.
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- 2023
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8. A magnetic nanoparticle-based microfluidic device fabricated using a 3D-printed mould for separation of Escherichia coli from blood.
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Jóskowiak A, Nogueira CL, Costa SP, Cunha AP, Freitas PP, and Carvalho CM
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- Humans, Escherichia coli, Lab-On-A-Chip Devices, Printing, Three-Dimensional, Magnetite Nanoparticles, Sepsis
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Herein, A microfluidic device is described, produced with a 3D-printed master mould that rapidly separates and concentrates Escherichia coli directly from whole blood samples, enabling a reduction in the turnaround time of bloodstream infections (BSIs) diagnosis. Moreover, it promotes the cleansing of the blood samples whose complexity frequently hampers bacterial detection. The device comprises a serpentine mixing channel with two inlets, one for blood samples (spiked with bacteria) and the other for magnetic nanoparticles (MNPs) functionalized with a (bacterio)phage receptor-binding protein (RBP) with high specificity for E. coli. After the magnetic labelling of bacteria throughout the serpentine, the microchannel ends with a trapping reservoir where bacteria-MNPs conjugates are concentrated using a permanent magnet. The optimized sample preparation device successfully recovered E. coli (on average, 66%) from tenfold diluted blood spiked within a wide range of bacterial load (10
2 CFU to 107 CFU mL-1 ). The non-specific trapping, tested with Staphylococcus aureus, was at a negligible level of 12%. The assay was performed in 30 min directly from diluted blood thus presenting an advantage over the conventional enrichment in blood cultures (BCs). The device is simple and cheap to fabricate and can be tailored for multiple bacterial separation from complex clinical samples by using RBPs targeting different species. Moreover, the possibility to integrate a biosensing element to detect bacteria on-site can provide a reliable, fast, and cost-effective point-of-care device., (© 2023. The Author(s).)- Published
- 2023
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9. Antiviral and Antibacterial Cold Spray Coating Application on Rubber Substrate, Disruption in Disease Transmission Chain.
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Saha DC, Boegel SJ, Tanvir S, Nogueira CL, Aucoin MG, Anderson WA, and Jahed H
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The objective of this study was to prepare a copper-coated rubber surface using cold spray technology with improved virucidal and antimicrobial properties to fight against highly transmissible viruses and bacteria. A successful cold spray coating was produced using irregular-shaped pure Cu powder on an escalator handrail rubber. The powder particles and the deposited coatings (single and double pass) were characterized in terms of particle morphology and size distribution, coating surface and coat/substrate cross-section properties. The bonding between powder and rubber surfaces was purely mechanical interlocking. The Cu powder penetration depth within the rubber surface increases with a number of depositions pass. The virucidal properties of the coated surface were tested utilizing surrogates for SARS-CoV-2: HCoV-229E, a seasonal human coronavirus, and baculovirus, a high-titer enveloped insect cell virus. A double-pass coated surface showed significant baculovirus inactivation relative to a bare rubber control surface after 2-h (approximately 1.7-log) and 4-h (approximately 6.2-log), while a 4-h exposure reduced HCoV-229E titer to below the limit of detection. A similar microbial test was performed using E. coli, showing a 4-log microbial reduction after 2-h exposure relative to the bare rubber. These promising results open a new application for cold spray in the health sector., Supplementary Information: The online version contains supplementary material available at 10.1007/s11666-023-01553-x., (© ASM International 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)
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- 2023
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10. The impact of a rub and rinse regimen on removal of human coronaviruses from contemporary contact lens materials.
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Nogueira CL, Boegel SJ, Shukla M, Ngo W, Jones L, and Aucoin MG
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- Humans, Contact Lens Solutions pharmacology, Methacrylates, Contact Lenses, Hydrophilic, Coronavirus
- Abstract
Purpose: To assess the influence of contemporary contact lens (CL) materials on human coronavirus attachment and the influence of a rub and rinse step to remove these viruses., Methods: The binding rates of HCoV-229E and HCoV-OC43 to eight soft CL materials and four rigid gas permeable materials were analyzed. The impact of a rub and rinse step to remove these viruses from all materials was examined. The efficacy of Biotrue (Bausch & Lomb), OPTI-FREE Puremoist (Alcon), Clear Care (Alcon) and cleadew (Ophtecs) to remove virus contamination from two representative soft lens materials (etafilcon A and lotrafilcon B) was also determined., Results: Approximately 10
2 to 103 infectious viral particles were recovered from each CL material. Although some materials were more prone to coronavirus adhesion, contamination of both viral types was reduced to below the limit of quantification (LQ) from all materials using a simple saline rinse step. Exposure to Clear Care and cleadew reduced the number of infectious viral particles from both etafilcon A and lotrafilcon B to below the LQ, while for Biotrue and OPTI-FREE Puremoist, infectious viral particles were reduced to below the LQ only when additional rub and rinse steps were included., Conclusion: Human coronavirus contamination can be easily removed from CL surfaces. Although CL care products containing hydrogen peroxide and povidone-iodine efficiently removed virus contamination from CL surfaces without the need for a rub and rinse step, a full regimen including rub and rinse steps is crucial when using CL care products based on non-oxidative systems., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: William Ngo, Lyndon Jones and Manish Shukla are members of the Centre for Ocular Research & Education (CORE) at the University of Waterloo. Over the past three years, members of CORE have received research funding from Alcon, Allergan, Allied Innovations, Aurinia Pharma, Azura Ophthalmics, Bausch Health Canada, BHVI, CooperVision, GL Chemtec, i-Med Pharma, J&J Vision, Lubris, Menicon, Nature’s Way, Novartis, Ophtecs, Oté Pharma, PS Therapy, Santen, SightGlass, SightSage, Topcon and Visioneering. Lyndon Jones is a consultant and/or serves on an advisory board for Alcon, CooperVision, J&J Vision, Novartis and Ophtecs. William Ngo has received consulting fees from Alcon., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)- Published
- 2022
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11. Antimicrobial polymeric composites for high-touch surfaces in healthcare applications.
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Liu M, Bauman L, Nogueira CL, Aucoin MG, Anderson WA, and Zhao B
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Antimicrobial polymer composites have long been utilized in the healthcare field as part of the first line of defense. These composites are desirable in that they pose a minimal risk of developing contagions with antibiotic resistance. For this reason, the field of antimicrobial composites has seen steady growth over recent years and is becoming increasingly important during the current COVID-19 pandemic. In this article, we first review the need of the antimicrobial polymers in high tough surfaces, the antimicrobial mechanism, and then the recent advances in the development of antimicrobial polymer composite including the utilization of intrinsic antimicrobial polymers, the addition of antimicrobial additives, and new exploration of surface patterning. While there are many established and developing methods of imbuing a material with antimicrobial activity, there currently is no standard quantification method for these properties leading to difficulty comparing the efficacy of these materials within the literature. A discussion of the common antimicrobial characterization methods is provided along with highlights on the need of a standardized quantification of antiviral and antibacterial properties in testing to allow ease of comparison between generated libraries and to facilitate proper screening. We also discuss and comment on the current trends of the development of antimicrobial polymer composites with long-lasting and specific antimicrobial activities, nontoxic properties, and environmental friendliness against a broad-spectrum of microbes., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article., (© 2022 Elsevier Inc. All rights reserved.)
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- 2022
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12. Exploitation of a Klebsiella Bacteriophage Receptor-Binding Protein as a Superior Biorecognition Molecule.
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Nogueira CL, Pires DP, Monteiro R, Santos SB, and Carvalho CM
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- Carrier Proteins, Humans, Klebsiella, Klebsiella pneumoniae, Bacteriophage Receptors, Klebsiella Infections
- Abstract
Klebsiella pneumoniae is a Gram-negative bacterium that has become one of the leading causes of life-threatening healthcare-associated infections (HAIs), including pneumonia and sepsis. Moreover, due to its increasingly antibiotic resistance, K. pneumoniae has been declared a global top priority concern. The problem of K. pneumoniae infections is due, in part, to the inability to detect this pathogen rapidly and accurately and thus to treat patients within the early stages of infections. The success in bacterial detection is greatly dictated by the biorecognition molecule used, with the current diagnostic tools relying on expensive probes often lacking specificity and/or sensitivity. (Bacterio)phage receptor-binding proteins (RBPs) are responsible for the recognition and adsorption of phages to specific bacterial host receptors and thus present high potential as biorecognition molecules. In this study, we report the identification and characterization of a novel RBP from the K. pneumoniae phage KpnM6E1 that presents high specificity against the target bacteria and high sensitivity (80%) to recognize K. pneumoniae strains. Moreover, adsorption studies validated the role of gp 86 in the attachment to bacterial receptors, as it highly inhibits (86%) phage adsorption to its Klebsiella host. Overall, in this study, we unravel the role and potential of a novel Klebsiella phage RBP as a powerful tool to be used coupled with analytical techniques or biosensing platforms for the diagnosis of K. pneumoniae infections.
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- 2021
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13. Aedes fluviatilis cell lines as new tools to study metabolic and immune interactions in mosquito-Wolbachia symbiosis.
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Conceição CC, da Silva JN, Arcanjo A, Nogueira CL, de Abreu LA, de Oliveira PL, Gondim KC, Moraes B, de Carvalho SS, da Silva RM, da Silva Vaz I Jr, Moreira LA, and Logullo C
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- Aedes immunology, Aedes metabolism, Animals, Cell Line, Female, Host Microbial Interactions immunology, Symbiosis immunology, Aedes microbiology, Immunity, Innate, Wolbachia immunology
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In the present work, we established two novel embryonic cell lines from the mosquito Aedes fluviatilis containing or not the naturally occurring symbiont bacteria Wolbachia, which were called wAflu1 and Aflu2, respectively. We also obtained wAflu1 without Wolbachia after tetracycline treatment, named wAflu1.tet. Morphofunctional characterization was performed to help elucidate the symbiont-host interaction in the context of energy metabolism regulation and molecular mechanisms of the immune responses involved. The presence of Wolbachia pipientis improves energy performance in A. fluviatilis cells; it affects the regulation of key energy sources such as lipids, proteins, and carbohydrates, making the distribution of actin more peripheral and with extensions that come into contact with neighboring cells. Additionally, innate immunity mechanisms were activated, showing that the wAflu1 and wAflu1.tet cells are responsive after the stimulus using Gram negative bacteria. Therefore, this work confirms the natural, mutually co-regulating symbiotic relationship between W. pipientis and A. fluviatilis, modulating the host metabolism and immune pathway activation. The results presented here add important resources to the current knowledge of Wolbachia-arthropod interactions., (© 2021. The Author(s).)
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- 2021
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14. Bacteriophage-receptor binding proteins for multiplex detection of Staphylococcus and Enterococcus in blood.
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Santos SB, Cunha AP, Macedo M, Nogueira CL, Brandão A, Costa SP, Melo LDR, Azeredo J, and Carvalho CM
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- Animals, Bacteriophage Receptors chemistry, Bacteriophage Receptors metabolism, Horses, Limit of Detection, Bacteremia blood, Bacteremia diagnosis, Bacteriophages genetics, Enterococcus chemistry, Enterococcus metabolism, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Staphylococcus chemistry, Staphylococcus metabolism, Viral Proteins chemistry, Viral Proteins genetics, Viral Proteins metabolism
- Abstract
Healthcare-associated infections (HCAIs) affect hundreds of millions of patients, representing a significant burden for public health. They are usually associated to multidrug resistant bacteria, which increases their incidence and severity. Bloodstream infections are among the most frequent and life-threatening HCAIs, with Enterococcus and Staphylococcus among the most common isolated pathogens. The correct and fast identification of the etiological agents is crucial for clinical decision-making, allowing to rapidly select the appropriate antimicrobial and to prevent from overuse and misuse of antibiotics and the consequent increase in antimicrobial resistance. Conventional culture methods are still the gold standard to identify these pathogens, however, are time-consuming and may lead to erroneous diagnosis, which compromises an efficient treatment. (Bacterio)phage receptor binding proteins (RBPs) are the structures responsible for the high specificity conferred to phages against bacteria and thus are very attractive biorecognition elements with high potential for specific detection and identification of pathogens. Taking into account all these facts, we have designed and developed a new, fast, accurate, reliable and unskilled diagnostic method based on newly identified phage RBPs and spectrofluorometric techniques that allows the multiplex detection of Enterococcus and Staphylococcus in blood samples in less than 1.5 hr after an enrichment step., (© 2020 Wiley Periodicals LLC.)
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- 2020
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15. Molecular epidemiology of Mycobacterium tuberculosis strains from prison populations in Santa Catarina, Southern Brazil.
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Medeiros TF, Nogueira CL, Prim RI, Scheffer MC, Alves EV, Rovaris DB, Zozio T, Rastogi N, and Bazzo ML
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- Bacterial Typing Techniques, Brazil epidemiology, Evolution, Molecular, Female, Genotype, Humans, Male, Molecular Epidemiology, Mycobacterium tuberculosis classification, Phylogeny, Public Health Surveillance, Tuberculosis diagnosis, Mycobacterium tuberculosis genetics, Prisoners, Tuberculosis epidemiology, Tuberculosis microbiology
- Abstract
The Tuberculosis (TB) notification rates are 5 to 81 times higher in prisons worldwide when compared to the general population. The state of Santa Catarina (SC) has few epidemiological data regarding TB in prisons. The aim of this study was to evaluate the molecular epidemiology of circulating strains in prisons of SC. The study comprised 95 clinical samples from six prisons. Among the cases included, all subjects were male, predominantly caucasians, and young adults, with low education level. The positive smear in the TB diagnosis comprised 62.0% of cases. About 50% of subjects had some condition associated with TB. The Spoligotyping results showed that the most frequent lineages were LAM (50.7%), T (22.2%) and S (11.6%). The 12-loci MIRU generated 62 different genotypes. The MSTs showed evolutionary relationships between Mycobacterium tuberculosis spoligotypes from SC and evolutionary relationships between the prison isolates and studied parameters. This first study on TB in prison units of SC highlighted the predominance of SIT216/LAM5, and SIT34/S. Interestingly, his profile was found to be different from that observed in a previous study performed with the state's general population. This data shows the need for continued surveillance of episodes of TB occurring among prison inmates in an emerging country like Brazil., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2018
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16. Identification of the Infection Source of an Outbreak of Mycobacterium Chelonae Keratitis After Laser in Situ Keratomileusis.
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Nascimento H, Viana-Niero C, Nogueira CL, Martins Bispo PJ, Pinto F, de Paula Pereira Uzam C, Matsumoto CK, Oliveira Machado AM, Leão SC, Höfling-Lima AL, and de Freitas D
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- Adult, Corneal Ulcer microbiology, Electrophoresis, Gel, Pulsed-Field, Eye Infections, Bacterial microbiology, Female, Humans, Lasers, Excimer therapeutic use, Male, Microbial Sensitivity Tests, Middle Aged, Mycobacterium Infections, Nontuberculous microbiology, Retrospective Studies, Corneal Ulcer epidemiology, Disease Outbreaks, Eye Infections, Bacterial epidemiology, Keratomileusis, Laser In Situ, Mycobacterium Infections, Nontuberculous epidemiology, Mycobacterium chelonae isolation & purification, Water Microbiology
- Abstract
Purpose: Nontuberculous mycobacteria keratitis is a rare but challenging complication of laser in situ keratomileusis (LASIK). This study was conducted to determine the source(s) of infection in a cluster of cases of keratitis after LASIK and to describe this outbreak and patients' outcomes., Methods: In this retrospective, case series, single-center study, 86 patients were included who underwent LASIK or photorefractive keratectomy between December 2011 and February 2012. Corneal scrapes from the affected eyes, samples of tap and distilled water, water from the reservoir of the distilling equipment, steamer, and autoclave cassette; antiseptic and anesthetic solutions and surgical instrument imprints were cultivated in liquid and on solid media. Gram-negative bacteria and yeasts were identified using automated systems and mycobacteria by polymerase chain reaction-restriction enzyme analysis of the hsp65 gene (PRA-hsp65) and DNA sequencing. Mycobacterial isolates were typed by pulsed-field gel electrophoresis. The cases and outcomes are described. The main outcome measure was identification of the source(s) of the mycobacterial infections., Results: Eight (15 eyes) of 86 patients (172 eyes) who underwent LASIK developed infections postoperatively; no patients who underwent photorefractive keratectomy developed infections. Mycobacterium chelonae was isolated from 4 eyes. The distilled water collected in the surgical facility contained the same M. chelonae strain isolated from the patients' eyes. Different gram-negative bacteria and yeasts were isolated from samples collected at the clinic but not from the patients' eyes., Conclusions: Tap water distilled locally in surgical facilities may be a source of infection after ocular surgery and its use should be avoided.
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- 2018
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17. Characterization of Mycobacterium chelonae -Like Strains by Comparative Genomics.
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Nogueira CL, de Almeida LGP, Menendez MC, Garcia MJ, Digiampietri LA, Chimara E, Cnockaert M, Palomino JC, Portaels F, Martin A, Vandamme P, and Leão SC
- Abstract
Isolates of the Mycobacterium chelonae - M. abscessus complex are subdivided into four clusters (CHI to CHIV) in the INNO-LiPA® Mycobacterium spp DNA strip assay. A considerable phenotypic variability was observed among isolates of the CHII cluster. In this study, we examined the diversity of 26 CHII cluster isolates by phenotypic analysis, drug susceptibility testing, whole genome sequencing and single-gene analysis. Pairwise genome comparisons were performed using several approaches, including average nucleotide identity (ANI) and genome-to-genome distance (GGD) among others. Based on ANI and GGD the isolates were identified as M. chelonae (14 isolates), M. franklinii (2 isolates) and M. salmoniphium (1 isolate). The remaining 9 isolates were subdivided into three novel putative genomospecies. Phenotypic analyses including drug susceptibility testing, as well as whole genome comparison by TETRA and delta differences, were not helpful in separating the groups revealed by ANI and GGD. The analysis of standard four conserved genomic regions showed that rpoB alone and the concatenated sequences clearly distinguished the taxonomic groups delimited by whole genome analyses. In conclusion, the CHII INNO-LiPa is not a homogeneous cluster; on the contrary, it is composed of closely related different species belonging to the M. chelonae-M. abscessus complex and also several unidentified isolates. The detection of these isolates, putatively novel species, indicates a wider inner variability than the presently known in this complex.
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- 2017
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18. Genomic epidemiology of a national outbreak of post-surgical Mycobacterium abscessus wound infections in Brazil.
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Everall I, Nogueira CL, Bryant JM, Sánchez-Busó L, Chimara E, Duarte RDS, Ramos JP, Lima KVB, Lopes ML, Palaci M, Kipnis A, Monego F, Floto RA, Parkhill J, Leão SC, and Harris SR
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- Adaptation, Biological genetics, Bacterial Typing Techniques, Brazil epidemiology, Cross Infection, Disease Transmission, Infectious, Gene Deletion, Genes, Bacterial, Genomics, Humans, Mycobacterium abscessus classification, Mycobacterium abscessus isolation & purification, Phenotype, Phylogeny, Plasmids genetics, Whole Genome Sequencing, Disease Outbreaks, Mycobacterium Infections, Nontuberculous epidemiology, Mycobacterium Infections, Nontuberculous microbiology, Mycobacterium abscessus genetics, Surgical Wound Infection epidemiology, Surgical Wound Infection microbiology
- Abstract
An epidemic of post-surgical wound infections, caused by a non-tuberculous mycobacterium, has been on-going in Brazil. It has been unclear whether one or multiple lineages are responsible and whether their wide geographical distribution across Brazil is due to spread from a single point source or is the result of human-mediated transmission. 188 isolates, collected from nine Brazilian states, were whole genome sequenced and analysed using phylogenetic and comparative genomic approaches. The isolates from Brazil formed a single clade, which was estimated to have emerged in 2003. We observed temporal and geographic structure within the lineage that enabled us to infer the movement of sub-lineages across Brazil. The genome size of the Brazilian lineage was reduced relative to most strains in the three subspecies of Mycobacterium abscessus and contained a novel plasmid, pMAB02, in addition to the previously described pMAB01 plasmid. One lineage, which emerged just prior to the initial outbreak, is responsible for the epidemic of post-surgical wound infections in Brazil. Phylogenetic analysis indicates that multiple transmission events led to its spread. The presence of a novel plasmid and the reduced genome size suggest that the lineage has undergone adaptation to the surgical niche.
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- 2017
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19. First insight into the molecular epidemiology of Mycobacterium tuberculosis in Santa Catarina, southern Brazil.
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Nogueira CL, Prim RI, Senna SG, Rovaris DB, Maurici R, Rossetti ML, Couvin D, Rastogi N, and Bazzo ML
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- Alcoholism, Bacteriological Techniques, Brazil epidemiology, Coinfection, Comorbidity, Female, Genotype, HIV Infections epidemiology, Humans, Male, Molecular Diagnostic Techniques, Molecular Epidemiology, Mycobacterium tuberculosis pathogenicity, Phenotype, Risk Factors, Substance-Related Disorders epidemiology, Tuberculosis prevention & control, Tuberculosis transmission, Mycobacterium tuberculosis genetics, Tuberculosis epidemiology, Tuberculosis microbiology
- Abstract
Molecular epidemiology of Mycobacterium tuberculosis is useful for understanding disease transmission dynamics, and to establish strategic measures for TB control and prevention. The aim of this study was to analyze clinical, epidemiological and molecular characteristics of MTBC clinical isolates from Santa Catarina state, southern Brazil. During one-year period, 406 clinical isolates of MTBC were collected from Central Laboratory of Public Health and typed by spoligotyping. Demographic and clinical data were collected from the Brazilian National Mandatory Disease Reporting System. The majority of cases occurred in highest population densities regions and about 50% had some condition associated with TB. Among all isolates, 5.7% were MDR, which showed association with drug addiction. LAM was the most predominant lineage with 47.5%, followed by the T superfamily with 25.9% and Haarlem with 12.3%. The MST showed two major groups: the first was formed mainly by the LAM lineage and the second was mainly formed by the T and Haarlem lineages. Others lineages were distributed in peripheral positions. This study provides the first insight into the population structure of M. tuberculosis in SC State. Spoligotyping and other genotyping analyses are important to establish strategic measures for TB control and prevention., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2016
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20. Mycobacterium saopaulense sp. nov., a rapidly growing mycobacterium closely related to members of the Mycobacterium chelonae--Mycobacterium abscessus group.
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Nogueira CL, Whipps CM, Matsumoto CK, Chimara E, Droz S, Tortoli E, de Freitas D, Cnockaert M, Palomino JC, Martin A, Vandamme P, and Leão SC
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- Animals, Bacterial Typing Techniques, Base Composition, Brazil, Cornea microbiology, DNA, Bacterial genetics, DNA, Ribosomal Spacer genetics, Fatty Acids chemistry, Genes, Bacterial, Humans, Molecular Sequence Data, Mycobacterium genetics, Mycobacterium isolation & purification, Mycobacterium Infections microbiology, Mycobacterium chelonae, Nucleic Acid Hybridization, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Zebrafish microbiology, Mycobacterium classification, Phylogeny
- Abstract
Five isolates of non-pigmented, rapidly growing mycobacteria were isolated from three patients and,in an earlier study, from zebrafish. Phenotypic and molecular tests confirmed that these isolates belong to the Mycobacterium chelonae-Mycobacterium abscessus group, but they could not be confidently assigned to any known species of this group. Phenotypic analysis and biochemical tests were not helpful for distinguishing these isolates from other members of the M. chelonae–M.abscessus group. The isolates presented higher drug resistance in comparison with other members of the group, showing susceptibility only to clarithromycin. The five isolates showed a unique PCR restriction analysis pattern of the hsp65 gene, 100 % similarity in 16S rRNA gene and hsp65 sequences and 1-2 nt differences in rpoB and internal transcribed spacer (ITS) sequences.Phylogenetic analysis of a concatenated dataset including 16S rRNA gene, hsp65, and rpoB sequences from type strains of more closely related species placed the five isolates together, as a distinct lineage from previously described species, suggesting a sister relationship to a group consisting of M. chelonae, Mycobacterium salmoniphilum, Mycobacterium franklinii and Mycobacterium immunogenum. DNA–DNA hybridization values .70 % confirmed that the five isolates belong to the same species, while values ,70 % between one of the isolates and the type strains of M. chelonae and M. abscessus confirmed that the isolates belong to a distinct species. The polyphasic characterization of these isolates, supported by DNA–DNA hybridization results,demonstrated that they share characteristics with M. chelonae–M. abscessus members, butconstitute a different species, for which the name Mycobacterium saopaulense sp. nov. is proposed. The type strain is EPM10906T (5CCUG 66554T5LMG 28586T5INCQS 0733T).
- Published
- 2015
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21. Molecular profiling of drug resistant isolates of Mycobacterium tuberculosis in the state of Santa Catarina, southern Brazil.
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Prim RI, Schörner MA, Senna SG, Nogueira CL, Figueiredo AC, Oliveira JG, Rovaris DB, and Bazzo ML
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- Adult, Bacterial Proteins genetics, Bacterial Typing Techniques, Brazil, Female, Genotype, Humans, Male, Polymorphism, Restriction Fragment Length, Sequence Analysis, DNA, Antitubercular Agents pharmacology, DNA, Bacterial, Drug Resistance, Multiple, Bacterial genetics, Mutation genetics, Mycobacterium tuberculosis drug effects, Tuberculosis, Multidrug-Resistant microbiology
- Abstract
Drug resistance is a global threat and one of the main contributing factors to tuberculosis (TB) outbreaks. The goal of this study was to analyse the molecular profile of multidrug-resistant TB (MDR-TB) in the state of Santa Catarina in southern Brazil. Fifty-three MDR Mycobacterium tuberculosis clinical isolates were analysed by spoligotyping and a partial region of the rpoB gene, which is associated with rifampicin resistance (RMP-R), was sequenced. Some isolates were also distinguished by their mycobacterial interspersed repetitive units (MIRU). S531L was the most prevalent mutation found within rpoB in RMP-R isolates (58.5%), followed by S531W (20.8%). Only two MDR isolates showed no mutations within rpoB. Isolates of the Latin American Mediterranean (LAM) family were the most prevalent (45.3%) found by spoligotyping, followed by Haarlem (9.4%) and T (7.5%) families. SIT106 was found in 26.4% of isolates and all SIT106 isolates typed by MIRU-12 (5 out of 14) belong to MIT251. There was a high correlation between the S531W mutation and the LAM family mainly because all SIT2263 (LAM9) isolates carry this mutation. Among isolates with the S531W mutation in rpoB MIRU demonstrates a cluster formed by four isolates (SIT2263 and MIT163) and very similar profiles were observed between eight of the nine isolates. Better characterisation of TB isolates may lead to new ways in which to control and treat TB in this region of Brazil.
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- 2015
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22. Antimicrobial susceptibility of rapidly growing mycobacteria using the rapid colorimetric method.
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Ramis IB, Cnockaert M, von Groll A, Nogueira CL, Leão SC, Andre E, Simon A, Palomino JC, da Silva PE, Vandamme P, and Martin A
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- Humans, Mycobacterium growth & development, Mycobacterium isolation & purification, Mycobacterium Infections microbiology, Nontuberculous Mycobacteria drug effects, Nontuberculous Mycobacteria growth & development, Nontuberculous Mycobacteria isolation & purification, Time Factors, Anti-Bacterial Agents pharmacology, Microbial Sensitivity Tests methods, Mycobacterium drug effects
- Abstract
Drug susceptibility testing (DST) of rapidly growing mycobacteria (RGM) are recommended for guiding the antimicrobial therapy. We have evaluated the use of resazurin in Mueller-Hinton medium (MHR) for MIC determination of RGM and compared the results with those obtained with the reference standard broth microdilution in Mueller-Hinton (MH) and with the resazurin microtiter assay (REMA) in 7H9 broth. The MIC of eight drugs: amikacin (AMI), cefoxitin (FOX), ciprofloxacin (CIP), clarithromycin (CLA), doxycycline (DOX), linezolid (LZD), moxifloxacin (MXF) and trimethoprim-sulfamethoxazole (TMP-SMX) were evaluated against 76 RGM (18 species) using three methods (MH, MHR, and REMA) in a 96-well plate format incubated at 37 °C over 3-5 days. Results obtained in the MH plates were interpreted by the appearance of turbidity at the bottom of the well before adding the resazurin. MHR and 7H9-REMA plates were read by visual observation for a change in color from blue to pink. The majority of results were obtained at day 5 for MH and 1 day after for MHR and 7H9-REMA. However, the preliminary experiment on time to positivity results using the reference strain showed that the resazurin can be added to the MH at day 2 to produce the results at day 3, but future studies with large sets of strains are required to confirm this suggestion. A high level of agreement (kappa 1.000-0.884) was obtained between the MH and the MHR. Comparison of results obtained with 7H9-REMA, on the other hand, revealed several discrepancies and a lower level of agreement (kappa 1.000-0.111). The majority of the strains were resistant to DOX and TMP-SMX, and the most active antimicrobials for RGM were AMI and FOX. In the present study, MHR represented an excellent alternative for MIC determination of RGM. The results could be read reliably, more easily, and more quickly than with the classical MH method.
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- 2015
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23. Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria.
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Wildner LM, Bazzo ML, Liedke SC, Nogueira CL, Segat G, Senna SG, Schlindwein AD, Oliveira JG, Rovaris DB, Bonjardim CA, Kroon EG, and Ferreira PC
- Subjects
- Bacterial Proteins genetics, Bacterial Typing Techniques, DNA, Bacterial genetics, Humans, Mycobacterium Infections microbiology, Mycobacterium Infections, Nontuberculous microbiology, Mycobacterium tuberculosis classification, Nontuberculous Mycobacteria classification, Polymerase Chain Reaction, Electrophoretic Mobility Shift Assay, Mycobacterium tuberculosis isolation & purification, Nontuberculous Mycobacteria isolation & purification, RNA, Ribosomal, 16S genetics
- Abstract
The identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.
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- 2014
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24. Demonstration of plasmid-mediated drug resistance in Mycobacterium abscessus.
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Matsumoto CK, Bispo PJ, Santin K, Nogueira CL, and Leão SC
- Subjects
- Anti-Bacterial Agents pharmacology, Bacterial Typing Techniques methods, Brazil, DNA, Bacterial genetics, Humans, Microbial Sensitivity Tests methods, Mycobacterium drug effects, Mycobacterium Infections drug therapy, Drug Resistance genetics, Mycobacterium genetics, Mycobacterium Infections microbiology, Plasmids genetics
- Abstract
Plasmid-mediated kanamycin resistance was detected in a strain of Mycobacterium abscessus subsp. bolletii responsible for a nationwide epidemic of surgical infections in Brazil. The plasmid did not influence susceptibility to tobramycin, streptomycin, trimethoprim-sulfamethoxazole, clarithromycin, or ciprofloxacin. Plasmid-mediated drug resistance has not been described so far in mycobacteria.
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- 2014
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25. External quality assurance with dried tube specimens (DTS) for point-of-care syphilis and HIV tests: experience in an indigenous populations screening programme in the Brazilian Amazon.
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Benzaken AS, Bazzo ML, Galban E, Pinto IC, Nogueira CL, Golfetto L, Benzaken NS, Sollis KA, Mabey D, and Peeling RW
- Subjects
- Brazil, Dried Blood Spot Testing standards, Feasibility Studies, Humans, Mass Screening organization & administration, Program Evaluation, Reproducibility of Results, Dried Blood Spot Testing methods, HIV Infections diagnosis, Health Services, Indigenous organization & administration, Indians, South American, Mass Screening standards, Point-of-Care Systems standards, Syphilis diagnosis
- Abstract
Objectives: The availability of point-of-care (POC) tests for infectious diseases has revolutionised the provision of healthcare for remote rural populations without access to laboratories. However, quality assurance for POC tests has been largely overlooked. We have evaluated the use and stability of dry tube specimens (DTS) for External Quality Assurance (EQA) for HIV and syphilis screening in remote indigenous populations in the Amazon region of Brazil., Methods: All healthcare workers (HCWs) participating in the community-screening were trained. We used HIV and syphilis DTS panels developed by the reference laboratory, containing samples with negative and positive results at different antibody concentrations, for both infections. DTS panels were distributed to HCWs in the communities for reconstitution and testing using POC HIV and syphilis tests. The results of testing were sent to the reference laboratory for marking and remedial action taken where necessary., Results: In total 268 HCWs tested 1607 samples for syphilis and 1608 samples for HIV. Results from HCWs showed a concordance rate of 90% for syphilis and 93% for HIV (κ coefficients of 0.74 and 0.78, respectively) with reference laboratories. Most false negatives were in samples of very low antibody concentration. DTS syphilis specimens produced the expected results after storage at 2-8°C or at 18-24°C for up to 3 weeks., Conclusions: The results show that POC tests for syphilis and HIV give valid results in environments where traditional tests do not work, but errors in the interpretation of POC test results were identified by the EQA programme using DTS. EQA using DTS can help to improve the quality of screening programmes using POC tests in remote regions.
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- 2014
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26. Correction: The Detection and Sequencing of a Broad-Host-Range Conjugative IncP-1β Plasmid in an Epidemic Strain of Mycobacterium abscessus subsp. bolletii.
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Leão SC, Matsumoto CK, Carneiro A, Ramos RT, Nogueira CL, Junior JD, Lima KV, Lopes ML, Schneider H, Azevedo VA, and da Costa da Silva A
- Abstract
[This corrects the article on p. e60746 in vol. 8.].
- Published
- 2013
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27. Acute phase proteins for the diagnosis of bacterial infection and prediction of mortality in acute complications of cirrhosis.
- Author
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Lazzarotto C, Ronsoni MF, Fayad L, Nogueira CL, Bazzo ML, Narciso-Schiavon JL, de Lucca Schiavon L, and Dantas-Corrêa EB
- Subjects
- Adult, Aged, Area Under Curve, Bacterial Infections blood, Bacterial Infections diagnosis, Bacterial Infections immunology, Biomarkers blood, Calcitonin Gene-Related Peptide, Chi-Square Distribution, Female, Humans, Liver Cirrhosis blood, Liver Cirrhosis immunology, Male, Middle Aged, Patient Admission, Predictive Value of Tests, Prognosis, Prospective Studies, ROC Curve, Risk Factors, Time Factors, Up-Regulation, Bacterial Infections etiology, Bacterial Infections mortality, C-Reactive Protein analysis, Calcitonin blood, Liver Cirrhosis complications, Liver Cirrhosis mortality, Protein Precursors blood
- Abstract
Introduction: Bacterial infection is a frequent complication in patients with decompensated liver cirrhosis and is related to high mortality rates during follow-up of these individuals. We sought to evaluate the diagnostic value of C-reactive protein (CRP) and procalcitonin (PCT) in diagnosing infection and to investigate the relationship between these biomarkers and mortality after hospital admission., Material and Methods: Prospective study that included cirrhotic patients admitted to the hospital due to complications of the disease. The diagnostic accuracy of CRP and PCT for the diagnosis of infection was evaluated by estimating the sensitivity and specificity and by measuring the area under the receiver operating characteristics curve (AUROC)., Results: A total of 64 patients and 81 hospitalizations were analyzed during the study. The mean age was 54.31 ± 11.87 years with male predominance (68.8%). Significantly higher median CRP and PCT levels were observed among infected patients (P < 0.001). The AUROC of CRP and PCT for the diagnosis of infection were 0.835 ± 0.052 and 0.860 ± 0.047, respectively (P = 0.273). CRP levels > 29.5 exhibited sensitivity of 82% and specificity of 81% for the diagnosis of bacterial infection. Similarly, PCT levels > 1.10 showed sensitivity of 67% and specificity of 90%. Significantly higher levels of CRP (P = 0.026) and PCT (P = 0.001) were observed among those who died within three months after admission., Conclusion: CRP and PCT were reliable markers of bacterial infection in subjects admitted due to complications of liver cirrhosis and higher levels of these tests are related to short-term mortality in those patients.
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- 2013
28. IGF-I and IGFBP-3 serum levels in patients hospitalized for complications of liver cirrhosis.
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Ronsoni MF, Lazzarotto C, Fayad L, Silva MC, Nogueira CL, Bazzo ML, Narciso-Schiavon JL, Dantas-Corrêa EB, and Schiavon Lde L
- Subjects
- Adult, Aged, Bilirubin blood, Biomarkers blood, Creatinine blood, Cross-Sectional Studies, Female, Humans, Inpatients, International Normalized Ratio, Liver Cirrhosis diagnosis, Liver Function Tests, Luminescent Measurements, Male, Middle Aged, Partial Thromboplastin Time, Predictive Value of Tests, Serum Albumin analysis, Serum Albumin, Human, Severity of Illness Index, Hospitalization, Insulin-Like Growth Factor Binding Protein 3 blood, Insulin-Like Growth Factor I analysis, Liver Cirrhosis blood, Liver Cirrhosis complications
- Abstract
Background: IGF-I and IGFBP-3 are part of IGF system and, due to their predominantly hepatic synthesis, they seem to correlate with hepatic dysfunction intensity., Aims: To investigate the significance of IGF-I and IGFBP-3 in patients with decompensated liver disease., Material and Methods: Cross-sectional study that included cirrhotic patients admitted to hospital due to complications of the disease, in whom IGF-I and IGFBP-3 serum levels were measured by chemiluminescence., Results: Seventy-four subjects with a mean age of 53.1 ± 11.6 years were included in the study, 73% were males. IGF-I levels were positively correlated with IGFBP-3 and albumin, and negatively correlated with Child-Pugh, MELD, creatinine, INR and aPTT ratio. IGFBP-3 levels were positively correlated with IGF-I and albumin, and negatively correlated with Child-Pugh, MELD, creatinine, INR, total bilirubin and aPTT ratio. Significantly lower scores of IGF-I and IGFBP-3 were observed in patients with higher MELD values and higher Child-Pugh classes (P < 0.05)., Conclusions: In cirrhotic patients admitted to hospital due to complications of the disease, IGF-I and IGFBP-3 serum levels were associated with variables related to liver dysfunction and to more advanced liver disease. The levels of these markers seem to undergo little influence from other clinical and laboratory variables, therefore mainly reflecting hepatic functional status.
- Published
- 2013
29. The detection and sequencing of a broad-host-range conjugative IncP-1β plasmid in an epidemic strain of Mycobacterium abscessus subsp. bolletii.
- Author
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Leão SC, Matsumoto CK, Carneiro A, Ramos RT, Nogueira CL, Lima JD Jr, Lima KV, Lopes ML, Schneider H, Azevedo VA, and da Costa da Silva A
- Subjects
- Bacterial Typing Techniques, Blotting, Southern, Electrophoresis, Gel, Pulsed-Field, Escherichia coli, Molecular Sequence Data, Polymerase Chain Reaction, Mycobacterium genetics, Plasmids genetics
- Abstract
Background: An extended outbreak of mycobacterial surgical infections occurred in Brazil during 2004-2008. Most infections were caused by a single strain of Mycobacterium abscessus subsp. bolletii, which was characterized by a specific rpoB sequevar and two highly similar pulsed-field gel electrophoresis (PFGE) patterns differentiated by the presence of a ∼50 kb band. The nature of this band was investigated., Methodology/principal Findings: Genomic sequencing of the prototype outbreak isolate INCQS 00594 using the SOLiD platform demonstrated the presence of a 56,267-bp [corrected] circular plasmid, designated pMAB01. Identity matrices, genetic distances and phylogeny analyses indicated that pMAB01 belongs to the broad-host-range plasmid subgroup IncP-1β and is highly related to BRA100, pJP4, pAKD33 and pB10. The presence of pMAB01-derived sequences in 41 M. abscessus subsp. bolletii isolates was evaluated using PCR, PFGE and Southern blot hybridization. Sixteen of the 41 isolates showed the presence of the plasmid. The plasmid was visualized as a ∼50-kb band using PFGE and Southern blot hybridization in 12 isolates. The remaining 25 isolates did not exhibit any evidence of this plasmid. The plasmid was successfully transferred to Escherichia coli by conjugation and transformation. Lateral transfer of pMAB01 to the high efficient plasmid transformation strain Mycobacterium smegmatis mc(2)155 could not be demonstrated., Conclusions/significance: The occurrence of a broad-host-range IncP-1β plasmid in mycobacteria is reported for the first time. Thus, genetic exchange could result in the emergence of specific strains that might be better adapted to cause human disease.
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- 2013
- Full Text
- View/download PDF
30. A study on the short-term effect of cafeteria diet and pioglitazone on insulin resistance and serum levels of adiponectin and ghrelin.
- Author
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Colombo G, Bazzo ML, Nogueira CL, Colombo MD, Schiavon LL, and d'Acampora AJ
- Subjects
- Animals, Body Weight, Energy Intake, Male, Pioglitazone, Rats, Rats, Wistar, Adiponectin blood, Dietary Carbohydrates pharmacology, Dietary Fats pharmacology, Ghrelin blood, Insulin Resistance, Thiazolidinediones pharmacology
- Abstract
The interaction between ghrelin and adiponectin is still controversial. We investigated the effect of cafeteria diet and pioglitazone on body weight, insulin resistance, and adiponectin/ghrelin levels in an experimental study on male Wistar rats. The animals were divided into four groups of 6 rats each, and received balanced chow with saline (CHOW-O) or pioglitazone (CHOW-P), or a cafeteria diet with saline (CAFE-O) or pioglitazone (CAFE-P). The chow/cafeteria diets were administered for 35 days, and saline/pioglitazone (10 mg · kg body weight(-1) · day(-1)) was added in the last 14 days prior to euthanasia. CAFE-O animals had a higher mean final weight (372.5 ± 21.01 g) than CHOW-O (317.66 ± 25.11 g, P = 0.017) and CHOW-P (322.66 ± 28.42 g, P = 0.035) animals. Serum adiponectin levels were significantly higher in CHOW-P (55.91 ± 20.62 ng/mL) than in CHOW-O (30.52 ± 6.97 ng/mL, P = 0.014) and CAFE-O (32.54 ± 9.03 ng/mL, P = 0.027) but not in CAFE-P. Higher total serum ghrelin levels were observed in CAFE-P compared to CHOW-P animals (1.65 ± 0.69 vs 0.65 ± 0.36 ng/mL, P = 0.006). Likewise, acylated ghrelin levels were higher in CAFE-P (471.52 ± 195.09 pg/mL) than in CHOW-P (193.01 ± 87.61 pg/mL, P = 0.009) and CAFE-O (259.44 ± 86.36 pg/mL, P = 0.047) animals. In conclusion, a cafeteria diet can lead to a significant weight gain. Although CAFE-P animals exhibited higher ghrelin levels, this was probably related to food deprivation rather than to a direct pharmacological effect, possibly attenuating the increase in adiponectin levels.
- Published
- 2012
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31. Alternative sputum preparation to improve polymerase chain reaction assay for Mycobacterium tuberculosis detection.
- Author
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Nogueira CL, Wildner LM, Senna SG, Rovaris D, Gruner MF, Jakimiu AR, da Silva RM, and Bazzo ML
- Subjects
- Brazil, Humans, Predictive Value of Tests, Sensitivity and Specificity, Tuberculosis microbiology, Mycobacterium tuberculosis genetics, Polymerase Chain Reaction methods, RNA, Bacterial isolation & purification, RNA, Ribosomal, 16S isolation & purification, Ribotyping methods, Sputum microbiology, Tuberculosis diagnosis
- Abstract
Background: Tuberculosis (TB), one of the major airborne infectious bacterial diseases, remains an important health problem worldwide. It is estimated that there are 1700 new cases per year in Santa Catarina State, Brazil., Objective: To improve polymerase chain reaction (PCR) sensitivity in detecting Mycobacterium tuberculosis in sputum samples., Methods: This study proposed the use of glass beads as a modification of the routine protocol for sputum preparation used in the Laboratory of Molecular Biology and Mycobacteria at the Federal University of Santa Catarina, Florianópolis, Brazil. The study comprised 120 sputum samples, 60 of which were treated with the routine protocol, while 60 were treated with the modified protocol using glass beads., Results: Samples treated with the routine protocol had a sensitivity of 56.7% (95%CI 44.1-69.2) in 16S rRNA PCR and 81.7% (95%CI 71.9-91.5) in insertion sequence (IS) 6110 PCR, compared with culture. Samples treated with the modified protocol had a sensitivity of 73.3% (95%CI 62.1-84.5) in 16S rRNA PCR and 100% in IS6110 PCR., Conclusion: The modified protocol using glass beads greatly improved mycobacterial detection in sputum samples compared with the routine protocol.
- Published
- 2012
- Full Text
- View/download PDF
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