MRSA colonization in workers from different occupational environments: a one health approach perspective

Projeto “Occupational exposure of ambulance drivers to bioburden” (IPL/2020/BIOAmbuDrivers_ESTeSL).Projeto “Bacterial Bioburden assessment in the context of occupational exposure and animal health of swine productions” (IPL/2016/BBIOR-Health).FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.Staphylococcus aureus and particularly methicillin-resistant S. aureus (MRSA) infections are currently associated with extremely high morbidity and mortality rates worldwide. The global escalation in the development of antibiotic-resistant human pathogens and S. aureus's ability in developing new clones with the capacity to invade community settings leads to an urgent need to develop accurate and efficient assessments of S. aureus colonization in occupational settings, particularly those with increased risk of human and animal colonization and food contamination. Here we present cross-sectional studies with the aim to assemble crucial information regarding MRSA prevalence in workers from five different Portuguese occupational environments (bakeries, swineries (humans and animals), ambulance crews, veterinary clinics, and healthcare facilities). Our data demonstrated a high prevalence of S. aureus asymptomatic carriers among bakery workers (40%; 75% MSSA and 25% MRSA), swinery workers (54%; 8% MSSA and 46% MRSA), firefighters (48.5%; 24% MSSA and 21% MRSA) and healthcare workers (Study 1: 42.2%; 18.4% MSSA and 23.7% MRSA, Study 2: 43.3% MRSA). S. aureus prevalence in veterinary staff was 7.1% (MSSA), lower than the results obtained in control groups (33.3% S. aureus; MRSA 4% to 10%). The present study sustains the urge to develop an accurate and efficient assessment of S. aureus human and animal colonization, particularly in high-risk occupational settings, with proper guidelines and validated procedures in order to avoid potential hazardous health outcomes associated with bioaerosol exposure and associated infectious diseases.info:eu-repo/semantics/publishedVersio

Fermentando resistência: organização coletiva de mulheres militantes para a produção de cerveja artesanal no Rio de Janeiro

TCC (especialização) - Universidade Federal de Santa Catarina. Centro de Ciências Agrárias. Curso de Especialização em AgroecossistemasO número de cervejarias registradas no Brasil aumentou significativamente nos últimos anos. Iniciativas de coletivos de mulheres produtoras de cerveja artesanal vêem contribuindo para esse aumento e para desconstruir uma estruturação masculinizada da prática cervejeira. Nesse sentido, sob a perspectiva do Materialismo Histórico Dialético, buscou-se discutir os desafios e possibilidades do trabalho coletivo de mulheres militantes envolvidas na produção de cerveja artesanal. Utilizamos uma metodologia de natureza qualitativa, composta por pesquisa bibliográfica, aplicação de entrevista estruturada e elaboração de material formativo em forma de caderno pedagógico. Foi possível observar que as práticas dos coletivos estudados se deparam constantemente com os desafios apresentados por uma sociedade estruturada no patriarcado, lucro, racismo, na exploração das classes sociais e apropriação dos meios de produção. E, que o trabalho coletivo cooperado possui papel fundamental para a existência e resistência das iniciativas. A dificuldade no acesso a um material sistematizado e focado nas dúvidas e desafios iniciais apresentada pelas entrevistadas foi esteio da estruturação do caderno pedagógico que visa estimular e facilitar etapas de organização destes e novos coletivos de mulheres cervejeiras.The number of registered breweries in Brazil has increased significantly in recent years. Initiatives by collectives of women craft beer producers are contributing to this increase and to deconstruct a masculinized structure of the beer practice. In this sense, from the perspective of Historical Dialectical Materialism, were discussed the challenges and possibilities of the collective work of militant women involved in the production of craft beer. We use a qualitative methodology, consisting of bibliographic research, application of structured interviews and preparation of training material in the form of a pedagogical notebook. It was possible to observe that the practices of the collectives studied are constantly faced with the challenges presented by a society structured in patriarchy, profit, racism, in the exploitation of social classes and appropriation of the means of production. And, that the cooperative collective work plays a fundamental role for the existence and resistance of the initiatives. The difficulty in accessing a systematized and focused material on the initial doubts and challenges presented by the interviewees was the basis for structuring the training pedagogical notebook that aims to encourage and facilitate stages of organization of these and new collectives of women brewers

Avaliação do potencial de virulência de Staphylococcus aureus associados a infeções de pele e tecidos moles

A espécie Staphylococcus aureus causa infeções de pele e tecidos moles (SSTIs) em humanos e animais, apresentando vários fatores de virulência. Este trabalho teve como objetivo determinar a frequência de diversas características associadas a virulência em S. aureus associados a SSTIs em humanos ou animais de companhia, explorando a relação destas com fenótipos de resistência aos antibióticos como a resistência à meticilina (estirpes MRSA) e multirresistência (MDR). Avaliou-se ainda o potencial de virulência de estirpes representativas num modelo de infeção. Estudaram-se 89 estirpes de S. aureus associadas a SSTIs, isoladas de humanos (n=34) ou animais de companhia (n=55). A tipagem do agr e rastreio dos genes lukS-PV/lukF-PV, que codificam a leucocidina PVL, foram realizados por PCR. A produção de biofilme foi avaliada pelo método de violeta de cristal. O potencial de virulência de estirpes representativas foi avaliado num modelo de infeção em Galleria mellonella. A distribuição dos tipos de agr foi semelhante nas duas coleções, sendo os tipos agrI ou agrII predominantes. Os genes lukS-PV/lukF-PV foram detetados em 21% e 2% das estirpes de origem humana ou animal, respetivamente, estando associados às linhagens MRSA-ST8-agrI, e linhagens relacionadas com S. aureus suscetível à meticilina (MSSA) ST152-agrI, ST30-agrIII e ST121-agrIV. A produção de biofilme foi detetada em 64% das estirpes, sendo mais frequente em estirpes de origem animal do que humana, e associada a linhagens associadas exclusivamente a humanos (MRSA/MSSA-ST8-agrI), a animais (MSSA-ST398-agrI) ou a ambos (MRSA-ST22-agrI, MRSA/MSSA-ST5-agrII). Não se verificou relação estatística entre produção de biofilme, tipo de agr, presença de PVL ou fenótipos MRSA/MDR. O modelo de infeção em Galleria mellonella revelou que estirpes MRSA-ST22 produtoras de biofilme apresentam um potencial de virulência superior à estirpe de referência S. aureus RN4220. Este estudo destaca a relevância dos biofilmes em linhagens de S. aureus associadas a SSTIs em humanos ou animais de companhia, muitas das quais associadas a fenótipos MRSA e/ou MDR. Foi ainda demonstrado o potencial de virulência destas estirpes, realçando a necessidade de uma abordagem Uma Só Saúde no estudo das infeções causadas por S. aureus.Staphylococcus aureus is an important agent of skin and soft-tissue infections (SSTIs) in humans and animals, harboring several virulence factors. This work aims to determine the occurrence of relevant traits associated with virulence in SSTI-related S. aureus isolated from humans or pets, exploring their relation with methicillin resistance (MRSA) and multidrug (MDR) phenotypes, and ultimately evaluating the virulence potential of representative strains in an infection model. The study collection included eighty-nine S. aureus isolated from humans (n=34) or pets (n=55). agr typing and screening of PVL-encoding genes, lukS-PV/lukF-PV was performed by PCR. Biofilm production was evaluated by the crystal violet adhesion assay. The virulence potential of representative strains was evaluated in a Galleria mellonella infection model. A similar distribution of agr types was observed in both collections, with agrI and agrII detected in >95% of all strains. Carriage of lukS-PV/lukF-PV was detected in 21% of human and 2% of animal isolates, being associated with MRSA-ST8-agrI, methicillin-susceptible (MSSA) lineages ST152-agrI, ST30-agrIII and ST121-agrIV. Overall, biofilm production was detected in 64% of the strains, yet more frequent in animals than in humans. This trait was detected in lineages associated solely with humans (MRSA/MSSA-ST8-agrI), animals (MSSA-ST398-agrI) or both (MRSA-ST22-agrI, MRSA/MSSA-ST5-agrII). Biofilm production, agr type, PVL carriage or antimicrobial resistance phenotype (MRSA/MDR) were not statistically associated. The Galleria mellonella infection model revealed that biofilm-producing MRSA-ST22 strains presented a higher virulence potential than the reference strain S. aureus RN4220. This study highlights the relevance of biofilm in human and/or animal SSTI-related S. aureus lineages, many of which associated with MRSA and/or MDR phenotypes. The study also evidenced the virulence potential of these strains, strengthening the need for a One Health approach in the study of infections caused by S. aureus

Clonal lineages, antimicrobial resistance, and pvl carriage of staphylococcus aureus associated to skin and soft-tissue infections from ambulatory patients in Portugal

Staphylococcus aureus (S. aureus) is a leading cause of skin and soft-tissue infections (SSTIs) in the community. In this study, we characterized a collection of 34 S. aureus from SSTIs in ambulatory patients in Portugal and analyzed the presence of Panton–Valentine leucocidin (PVL)-encoding genes and antibiotic-resistance profile, which was correlated with genetic determinants, plasmid carriage, and clonal lineage. Nearly half of the isolates (15, 44.1%) were methicillin-resistant Staphylococcus aureus (MRSA) and/or multidrug resistant (MDR). We also detected resistance to penicillin (33/34, 97.1%), fluoroquinolones (17/34, 50.0%), macrolides and lincosamides (15/34, 44.1%), aminoglycosides (6/34, 17.6%), and fusidic acid (2/34, 5.9%), associated with several combinations of resistance determinants (blaZ, erm(A), erm(C), msr(A), mph(C), aacA-aphD, aadD, aph(3′ )-IIIa, fusC), or mutations in target genes (fusA, grlA/gyrA). The collection presented a high genetic diversity (Simpson’s index of 0.92) with prevalence of clonal lineages CC5, CC22, and CC8, which included the MRSA and also most MDR isolates (CC5 and CC22). PVL-encoding genes were found in seven isolates (20.6%), three methicillin-susceptible Staphylococcus aureus (MSSA) (ST152-agrI and ST30-agrIII), and four MRSA (ST8-agrI). Plasmid profiling revealed seventeen distinct plasmid profiles. This work highlights the high frequency of antimicrobial resistance and PVL carriage in SSTIs-related S. aureus outside of the hospital environment.publishersversionpublishe

Staphylococcus aureus causing skin and soft tissue infections in companion animals : antimicrobial resistance profiles and clonal lineages

Research Areas: Infectious Diseases ; Pharmacology & PharmacyABSTRACT - Staphylococcus aureus is a relevant agent of skin and soft tissue infections (SSTIs) in animals. Fifty-five S. aureus comprising all SSTI-related isolates in companion animals, collected between 1999 and 2018 (Lab 1) or 2017 and 2018 (Lab 2), were characterized regarding susceptibility to antibiotics and heavy metals and carriage of antimicrobial resistance determinants. Clonal lineages were established by PFGE, MLST and agr typing. Over half of the isolates (56.4%, 31/55) were methicillin-resistant S. aureus (MRSA), and 14.5% showed a multidrug resistance (MDR) phenotype. Resistance was most frequently observed for beta-lactams (81.8%, related to blaZ and/or mecA), fluoroquinolones (56.4%) and macrolides/lincosamides (14.5%, related to erm(A) or erm(C)). The distributions of heavy-metal MICs allowed the detection of non-wild-type populations associated with several resistance genes. The collection showed genetic diversity, with prevalence of clonal lineage ST22-agrI (45.5%, 25/55), comprising only MRSA isolates, and several less frequently detected clones, including ST5-agrII (14.6%, 8/55), ST398-agrI (9.1%, 5/55) and ST72-agrI (7.3%, 4/55). This work highlights the high frequency of SSTI-related MRSA strains that reflect the clonal lineages circulating both in companion animals and humans in Portugal, reinforcing the need for a One Health approach when studying staphylococci causing infections in companion animals.info:eu-repo/semantics/publishedVersio

Virulence potential of biofilm producing Staphylococcus pseudintermedius, Staphylococcus aureus and Staphylococcus coagulans causing skin infections in companion animals

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Role of DNA methylation in persistent diabetic macular edema

Projeto ID&CA 2021_IPL/2021/DiffMeDiME_ESTeSLBackground: a) Disease duration and metabolic control are insufficient to understand Diabetic Macular Edema (DME), the leading cause of vision loss in people with diabetes; b) 30-40% of cases of DME do not respond optimally to AVEGF (loading phase); c) poor genetic association in DR development (<25% RD) and PDR progression (25-50%). Purpose: To study the role of DNA methyltransferase expression (DNMT1,DNMT3a, DNMT3b) in persistent diabetic macular edema.info:eu-repo/semantics/publishedVersio

DNA methyltransferase expression (DNMT1, DNMT3a and DNMT3b) as a potential biomarker for anti-VEGF diabetic macular edema response

Funding Information: This project was partially supported by an IDI&CA grant IPL/2021/DiffMeDiME_ESTeSL by H&TRC- Health & Technology Research Center, ESTeSL- Escola Superior de Tecnologia da Saúde, Instituto Politécnico de Lisboa and by Retina Institute of Lisbon (IRL).Purpose: DNA methylation is involved in Diabetic Retinopathy progression showing a metabolic memory mechanism. However, the association of DNA methyltransferase with diabetic macular edema is still unknown. We aimed to describe the differences in DNA methyltransferase gene expression in patients with different diabetic macular edema responses. Methods: A total of 27 diabetic patients, aged 59–90 years, were prospectively enrolled in this cross-sectional study. The participants were classified into control group (CG, n = 11), diabetic macular edema responders (rDME, n = 9) and non-responder diabetic macular edema (nrDME, n = 7) after anti-vascular endothelial growth factor (anti-VEGF) treatment. Only cases with a complete ophthalmological examination, digital 133° color fundus, and SD-OCT assessments were used. After RNA extraction and first-strand cDNA synthesis, quantitative real-time PCR was performed with specific primers on the CFX Connect™ Real-Time PCR Detection System to assess differential transcriptional expression patterns. Results: The DNMT1 gene showed a positive correlation (r = 0.617; p = 0.043) with Best Corrected Visual Acuity (BCVA) in CG, a positive correlation (r = 0.917; p = 0.010) with HbA1c in nrDME and a negative correlation (r = −0.659; p = 0.049) with GCL-IPL thickness in rDME. DNMT3A gene showed a positive correlation (r = −0.890; p = 0.001) with Sub-foveal Choroidal thickness in rDME whereas DNMT3b gene showed a negative correlation (r = −0.815; p = 0.007) with HbA1c and RNFL (r = −0.664; p = 0.026) in CG. Conclusions: Patients with similar metabolic profile risk factors showed associated DNA methyltransferase transcriptional expression patterns differences fitting with the anti-VEGF diabetic macular edema response. Further studies are needed to clarify if these results (1) reflect disease evolution, (2) translate the therapeutic impact, (3) or can help to predict the therapeutic resistance profile.publishersversionepub_ahead_of_prin

DNA methylation in diabetic macular edema: first report

Projeto IPL/2021/DiffMeDiME_ESTeSLEnvelhecimento humano e estilos de vida tem contribuído para aumento da DM. Edema macular diabético (EMD), uma das manifestações oculares da RD, é a principal causa de perda de visão em pessoas diabéticas. 30-40% casos EMD não respondem da melhor forma aos AVEGF (loading phase). Resistência pode persistir após 12 meses tratamento (+/50%). Objetivo do estudo: Desenvolver uma caracterização multimodal não invasiva (SD OCT e OCT A) combinada com a avaliação da metilação de DNA de forma a pode esclarecer alguns mecanismos importantes da doença mas também para novas abordagens terapêuticas que contribuem para a medicina de precisão.info:eu-repo/semantics/publishedVersio

Analysis of the biological effect of Carica papaya leaf extracts on the viability and proliferation of K562 cells

Os autores reconhecem o apoio do H&TRC – Centro de Investigação em Saúde e Tecnologia, ESTeSL – Escola Superior de Tecnologia da Saúde de Lisboa, Instituto Politécnico de Lisboa (apoio FCT/MCTES, UIDB/05608/2020 e UIDP/05608/2020).Introdução – A anemia falciforme é uma doença monogénica causada por mutações no gene da β-globina que afeta a estrutura da hemoglobina, sendo associada a diversas complicações clínicas com elevadas taxas de morbilidade e mortalidade. A reativação farmacológica da hemoglobina fetal (HbF) por compostos como a hidroxiureia (HU) é um dos tratamentos atualmente disponíveis; contudo, o seu perfil de segurança e o elevado custo em países subdesenvolvidos limitam a sua utilização. Nesse contexto é essencial estudar novos compostos indutores da HbF com baixa citotoxicidade e que possam estar amplamente disponíveis, como é o caso de extratos de folhas da Carica papaya (CP), uma planta medicinal com propriedades antioxidantes e anti-inflamatórias. Objetivos – Este estudo pretende avaliar o efeito do extrato metanólico das folhas de CP (EMFCP) em parâmetros biológicos como a proliferação e a viabilidade celular em células K562. Método – As células K562 foram expostas durante 72h ao EMFCP a 500 µg/mL e durante 24 horas ao EMFCP (0,5; 50; 100 µg/mL) e à HU (25 μg/mL). A proliferação e viabilidade celular foram analisadas através da quantificação celular pelo método de exclusão do azul de tripano. Resultados – Os resultados demonstram que a proliferação e a viabilidade celular foram afetadas pelo EMFCP apenas na concentração de 500 µg/mL, não se tendo verificado alteração nestes parâmetros nas restantes concentrações utilizadas. Conclusão – Os resultados mostraram que os EMFCP não são citotóxicos quando incubados em células K562 em concentrações inferiores ou iguais a 100 μg/mL, permitindo assim explorar este composto na avaliação do seu potencial terapêutico no contexto da anemia falciforme.ABSTRACT: Background – Sickle cell disease (SCD) is a genetic blood disorder caused by mutations in the β-globin gene that affects the shape and transport of red blood cells in blood vessels, leading to various clinical complications. The pharmacological reactivation of fetal hemoglobin (HbF) through compounds such as hydroxyurea (HU), is one of the currently available treatments, however, their safety concerns and expensive cost in low- and middle-Income countries limit their use. In this context, it is essential to study novel HbF-inducing compounds that have scarcer adverse effects and can be widely available, such as Carica papaya leaf (CP) extracts, a medicinal plant with anti-oxidant and anti-inflammatory properties. Aim of the study – Evaluate the effect of CP methanolic leaf extracts (CPMLE) on biological parameters such as cell proliferation and viability on the K562 cell line. Methods – K562 cells were exposed for 72 hours to CPMLE at 500 µg/mL and for 24 hours to CPMLE (0.5; 50; 100 µg/mL) and HU (25 μg/mL). Cell proliferation and viability after CPMLE exposure were analyzed by cell counting. Results – The results demonstrate that cell proliferation and viability were affected by CPMLE only at the concentration of 500 µg/mL, with no effects being observed at the lower concentrations analysed. Conclusion – Our results showed that CPMLE is not cytotoxic when incubated at concentrations equal to or below 100 μg/mL thus allowing us to explore this compound in the evaluation of its therapeutic potential, in the context of sickle cell anemia treatment.Este estudo foi financiado pelo projeto IDI&CA-IPL/2021/EpiCa/ESTeSL.info:eu-repo/semantics/publishedVersio