Inconsistencies in conventional culture vs molecular approaches: unveiling distinct dynamics in cystic fibrosis polymicrobial communities

Book of Abstracts of CEB Annual Meeting 2017info:eu-repo/semantics/publishedVersio

An in vitro dynamic model of catheter-associated urinary tract infections to investigate the role of uncommon bacteria on the Escherichia coli microbial consortium

About 9% of nosocomial infections are attributed to catheter-associated urinary tract infections (CAUTIs). Uncommon bacteria (Delftia tusurhatensis) have been isolated in CAUTIs in combination with wellestablished pathogenic bacteria such as E. coli. Nonetheless, the reason why E. coli coexists with other bacteria instead of outcompeting and completely eliminating them are unknown. As such, a flow cell reactor simulating the hydrodynamic conditions found in CAUTIs (shear rate of 15 s-1) was used to characterize the microbial physiology of E. coli and D. tsuruhatensis individually and in consortium, in terms of growth kinetics and substrate uptake. Single-species biofilms showed that up to 48 h the CFU counts significantly increased for both species (p<0.05). After 48 h, both species stabilized with similar CFU values reaching log 6.24 CFU.cm2 for E. coli and log 6.31 CFU.cm2 for D. tsuruhatensis (p>0.05). The assessment of spatial distribution of dual-species biofilms by LNA/2´OMe-FISH revealed that E. coli and D. tsuruhatensis coexist and tend to co-aggregate over time, which implies that bacteria are able to cooperate synergistically. Substrate uptake measurements revealed that in artificial urine medium the bacteria metabolized lactic acid, uric acid (E. coli and D. tsuruhatensis) and citric acid (D. tsuruhatensis). In the consortium, D. tsuruhatensis consumed citric acid more rapidly, presumably leaving more uric acid available in the medium to be used by E. coli. In conclusion, metabolic cooperation between E. coli and uncommon species seems to occur when these species share the same environment, leading to the formation of a stable microbial community

Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes

Supplementary data to this article can be found online at https:// doi.org/10.1016/j.fm.2018.12.009.Listeria monocytogenes is one of the most important foodborne pathogens due to the high hospitalization and mortality rates associated to an outbreak. Several new molecular methods that accelerate the identification of L. monocytogenes have been developed, however conventional culture-based methods still remain the gold standard. In this work we developed a novel Peptide Nucleic Acid Fluorescence in situ Hybridization (PNA-FISH) method for the specific detection of L. monocytogenes. The method was based on an already existing PNA probe, LmPNA1253, coupled with a novel blocker probe in a 1:2 ratio. The method was optimized for the detection of L. monocytogenes in food samples through an evaluation of several rich and selective enrichment broths. The best outcome was achieved using One Broth Listeria in a two-step enrichment of 24h plus 18h. For validation in food samples, ground beef, ground pork, milk, lettuce and cooked shrimp were artificially contaminated with two ranges of inoculum: a low level (0.22CFU/25g or mL) and a high level (210CFU/25g or mL). The PNA-FISH method performed well in all types of food matrices, presenting an overall accuracy of 99% and a detection limit of 0.5CFU/25g or mL of food sample.Nuno R. da Silva acknowledges the financial support byFundac¸ ão para a Ciência e Tecnologia (FCT) of the Portuguese’s Min-istry for Science, Technology and Higher Education (MCTES), in theframework of the POCH – Programa Operacional Capital Humano,co-funded by the European Social Fund (SFRH/BD/111825/2015).This work was also supported by FCT under the scope ofthe strategic funding of UID/BIO/04469 unit and COMPETE2020 (POCI-01-0145-FEDER-006684) and BioTecNorte opera-tion (NORTE-01-0145-FEDER-000004) funded by the EuropeanRegional Development Fund under the scope of Norte2020 – Pro-grama Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Agent-based model of diffusion of N-acyl homoserine lactones in a multicellular environment of Pseudomonas aeruginosa and Candida albicans

Experimental incapacity to track microbemicrobe interactions in structures like biofilms, and the complexity inherent to the mathematical modelling of those interactions, raises the need for feasible, alternative modelling approaches. This work proposes an agent-based representation of the diffusion of N-acyl homoserine lactones (AHL) in a multicellular environment formed by Pseudomonas aeruginosa and Candida albicans. Depending on the spatial location, C. albicans cells were variably exposed to AHLs, an observation that might help explain why phenotypic switching of individual cells in biofilms occurred at different time points. The simulation and algebraic results were similar for simpler scenarios, although some statistical differences could be observed (p<0.05). The model was also successfully applied to a more complex scenario representing a small multicellular environment containing C. albicans and P. aeruginosa cells encased in a 3-D matrix. Further development of this model may help create a predictive tool to depict biofilm heterogeneity at the single-cell level.This work has been funded by a Research Grant 2014 by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) to AL; the Portuguese Foundation for Science and Technology (FCT) [grant numbers UID/ BIO/04469/2013, UID/EQU/00511/2013] units and COMPETE 2020 [grant numbers POCI-01-0145-FEDER-006684, POCI-01-0145-FEDER-006939]; North Portugal Regional Operational Programme (NORTE 2020) [grant number NORTE‐01‐0145‐FEDER‐000005 – LEPABE-2-ECO-INNOVATION] under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).info:eu-repo/semantics/publishedVersio

uma análise transversal

Publisher Copyright: Copyright © Ordem dos Médicos 2022.Introduction: Pre-exposure prophylaxis (PrEP) has gained relevance as a method of prevention for HIV in certain people and settings. Following the publication of the guideline on PrEP prescribing in Portugal, we aimed to assess the knowledge of Portuguese Medical Students about PrEP. Material and Methods: An online survey was sent to Medical students of Portuguese Medical Schools. We conducted a descriptive analysis of the results and an analytic cross-sectional study to identify factors associated with “knowing about PrEP”, “having had one class about PrEP” and “identifying eligible groups correctly”. Results: Of the 796 students that responded to the survey, 64.6% were aware of what PrEP is. Of these, 34.44% acquired this knowledge during their training. Out of the total amount of respondents, 4.77% could identify correctly and completely the eligible groups for PrEP. As the training years went by, the probability of being aware of PrEP, having had one class about PrEP, and identifying the eligible groups correctly, increased. Of the sixth-year students, 43.48% had had one class about PrEP and among the students that were aware of PrEP, 28% identified what the eligible groups were. After adjusting for the school year, we found differences between Medical Schools regarding the outcomes. The association between the different ways of knowing about PrEP and the ability to correctly identify eligible groups for PrEP was not statistically significant. Conclusion: The differences between Medical Schools could be harmonized through changes in the medical curricula that would allow for this topic to be addressed more often.publishersversionpublishe

Targeting miR-9 in gastric cancer cells using locked nucleic acid oligonucleotides

Abstract Background Gastric cancer is the third leading cause of cancer-related mortality worldwide. Recently, it has been demonstrated that gastric cancer cells display a specific miRNA expression profile, with increasing evidence of the role of miRNA-9 in this disease. miRNA-9 upregulation has been shown to influence the expression of E-cadherin-encoding gene, triggering cell motility and invasiveness. Results In this study, we designed LNA anti-miRNA oligonucleotides with a complementary sequence to miRNA-9 and tested their properties to both detect and silence the target miRNA. We could identify and visualize the in vitro uptake of low-dosing LNA-based anti-miRNA oligonucleotides without any carrier or transfection agent, as early as 2 h after the addition of the oligonucleotide sequence to the culture medium. Furthermore, we were able to assess the silencing potential of miRNA-9, using different LNA anti-miRNA oligonucleotide designs, and to observe its subsequent effect on E-cadherin expression. Conclusions The administration of anti-miRNA sequences even at low-doses, rapidly repressed the target miRNA, and influenced the expression of E-cadherin by significantly increasing its levels

Characterisation of microbial attack on archaeological bone

As part of an EU funded project to investigate the factors influencing bone preservation in the archaeological record, more than 250 bones from 41 archaeological sites in five countries spanning four climatic regions were studied for diagenetic alteration. Sites were selected to cover a range of environmental conditions and archaeological contexts. Microscopic and physical (mercury intrusion porosimetry) analyses of these bones revealed that the majority (68%) had suffered microbial attack. Furthermore, significant differences were found between animal and human bone in both the state of preservation and the type of microbial attack present. These differences in preservation might result from differences in early taphonomy of the bones. © 2003 Elsevier Science Ltd. All rights reserved

Ciência, Crise e Mudança. 3.º Encontro Nacional de História das Ciências e da Tecnologia. ENHCT2012

III Encontro Nacional de História das Ciências e da Tecnologia. O Centro de Estudos de História e Filosofia da Ciência, organiza o 3.º Encontro Nacional de História da Ciência e da Técnica, sob o tema «Ciência, Crise e Mudança» que tem lugar na Universidade de Évora, nos dias 26, 27 e 28 de Setembro de 2012. O Primeiro Encontro Nacional de História da Ciência teve lugar em 21 e 22 Julho de 2009, no seguimento do programa de estímulo ao de¬senvolvimento da História da Ciência em Portugal e de valorização do património cultural e científico do País, lançado pelo Ministério da Ciência, Tecnologia e Ensino Superior (MCTES) em 31 de Janeiro desse ano. A sua organização coube a investigadores do Instituto de História Contemporânea (IHC), da FCSH da UNL, e do Centro Científico e Cultural de Macau (CCCM), em cujas instalações se realizou. De en¬tre as conclusões do Encontro, destacou-se a de realizar periodicamen¬te novos Encontros Nacionais, a serem organizados de forma rotativa por diferentes centros e núcleos de investigadores. Na sequência deste Primeiro Encontro, o Centro Interuniversitário de História das Ciências e da Tecnologia (CIUHCT) organizou, entre 26 e 28 de Julho de 2010, o II Encontro, dedicado ao tema “Comunicação das Ciências e da Tecnologia em Portugal: Agentes, Meios e Audiências”. Cabe agora ao CEHFCi cumprir o que foi decidido no final deste Encontro. Na situação económica e política que hoje vivemos torna-se particularmente urgente aprofundar o estudo e o debate sobre a interação entre a Sociedade, a Ciência e a sua História. Coordenação Científica e Executiva do encontro estiveram a cargo de dois investigadores CEHFCi: Maria de Fátima Nunes, José Pedro Sousa Dia