Environmental Footprints in Food Services: A Scoping Review.

The collective meals market generates significant revenue for the world economy. Food services are responsible for consuming large amounts of water and energy, as well as generating a substantial volume of waste, which is often improperly disposed of. Given the unchecked expansion of food services, the lack of proper management of environmental resources can undermine sustainability principles, posing a threat to future generations. This scoping review aimed to synthesize the existing scientific literature on carbon and water footprints in food services, describing the main methods and tools used and what strategies have been proposed to mitigate the high values of these footprints. The search for articles was performed on 6 June 2024 in seven electronic databases, using MeSH Terms and adaptations for each database from database inception. The search for local studies was complemented by a manual search in the list of references of the studies selected to compose this review. It included quantitative studies assessing footprints (water or carbon) in food services and excluded reviews, studies that reported footprints for diets, and protocols. A total of 2642 studies were identified, and among these, 29 were selected for this review. According to the findings, it was observed that meats, especially beef, contribute more to water and carbon footprint compared to other proteins. Mitigation strategies for the water footprint include promoting plant-based diets, menu changes, and awareness.

Effects of temperature up-shift and UV-A radiation on fatty acids content and expression of desaturase genes in cyanobacteria Microcystis aeruginosa: stress tolerance and acclimation responses.

Temperature up-shift and UV-A radiation effects on growth, lipid damage, fatty acid (FA) composition and expression of desaturase genes desA and desB were investigated in the cyanobacteria Microcystis aeruginosa. Although UV-A damaging effect has been well documented, reports on the interactive effects of UV radiation exposure and warming on cyanobacteria are scarce. Temperature and UV-A doses were selected based on the physiological responses previously obtained by studies with the same M. aeruginosa strain used in this study. Cells pre-grown at 26 °C were incubated at the same temperature or 29 °C and exposed to UV-A + PAR and only PAR for 9 days. Growth rate was significantly affected by UV-A radiation independently of the temperature throughout the experiment. High temperature produced lipid damage significantly higher throughout the experiment, decreasing at day 9 as compared to 26 °C. In addition, the cells grown at 29 °C under UV-A displayed a decrease in polyunsaturated FA (PUFA) levels, with ω3 PUFA being mostly affected at the end of exposure. Previously, we reported that UV-A-induced lipid damage affects differentially ω3 and ω6 PUFAs. We report that UV-A radiation leads to an upregulation of desA, possibly due to lipid damage. In addition, the temperature up-shift upregulates desA and desB regardless of the radiation. The lack of lipid damage for UV-A on ω3 could explain the lack of transcription induction of desB. The significant ω6 decrease at 26 °C in cells exposed to UV-A could be due to the lack of upregulation of desA.

AP-1γ2 is an adaptor protein 1 variant required for endosome-to-Golgi trafficking of the mannose-6-P receptor (CI-MPR) and ATP7B copper transporter.

Selective retrograde transport from endosomes back to the trans-Golgi network (TGN) is important for maintaining protein homeostasis, recycling receptors, and returning molecules that were transported to the wrong compartments. Two important transmembrane proteins directed to this pathway are the Cation-Independent Mannose-6-phosphate receptor (CI-MPR) and the ATP7B copper transporter. Among CI-MPR functions is the delivery of acid hydrolases to lysosomes, while ATP7B facilitates the transport of cytosolic copper ions into organelles or the extracellular space. Precise subcellular localization of CI-MPR and ATP7B is essential for the proper functioning of these proteins. This study shows that both CI-MPR and ATP7B interact with a variant of the clathrin adaptor 1 (AP-1) complex that contains a specific isoform of the γ-adaptin subunit called γ2. Through synchronized anterograde trafficking and cell-surface uptake assays, we demonstrated that AP-1γ2 is dispensable for ATP7B and CI-MPR exit from the TGN while being critically required for ATP7B and CI-MPR retrieval from endosomes to the TGN. Moreover, AP-1γ2 depletion leads to the retention of endocytosed CI-MPR in endosomes enriched in retromer complex subunits. These data underscore the importance of AP-1γ2 as a key component in the sorting and trafficking machinery of CI-MPR and ATP7B, highlighting its essential role in the transport of proteins from endosomes.

HIV-1 Nef Changes the Proteome of T Cells Extracellular Vesicles Depleting IFITMs and Other Antiviral Factors.

Extracellular vesicles (EVs) are biomolecule carriers for intercellular communication in health and disease. Nef is a HIV virulence factor that is released from cells within EVs and is present in plasma EVs of HIV-1 infected individuals. We performed a quantitative proteomic analysis to fully characterize the Nef-induced changes in protein composition of T cell-derived EVs and identify novel host targets of HIV. Several proteins with well-described roles in infection or not previously associated with HIV pathogenesis were specifically modulated by Nef in EVs. Among the downregulated proteins are the interferon-induced transmembrane 1, 2, and 3 (IFITM1-3) proteins, broad-spectrum antiviral factors known to be cell-to-cell transferable by EVs. We demonstrate that Nef depletes IFITM1-3 from EVs by excluding these proteins from the plasma membrane and lipid rafts, which are sites of EVs biogenesis in T cells. Our data establish Nef as a modulator of EVs' global protein content and as an HIV factor that antagonizes IFITMs.

Role of Pel and Psl polysaccharides in the response of Pseudomonas aeruginosa to environmental challenges: oxidative stress agents (UVA, H2O2, sodium hypochlorite) and its competitor Staphylococcus aureus.

Pseudomonas aeruginosa is a versatile bacterium capable of adapting to a wide range of stress factors, including solar UVA radiation (400-315 nm). High UVA doses produce lethal effects due to the action of reactive oxygen species. Sublethal UVA doses also induces oxidative damage, but, in addition, it triggers a variety of adaptive responses, including the overexpression of pelA and pslA genes in P. aeruginosa. These genes encode the synthesis of Pel and Psl, which are essential polysaccharides in biofilm formation. The present study analysed the role of Pel and Psl in the adaptive responses generated by exposure to low UVA doses, and their importance in the response to lethal doses of UVA, hydrogen peroxide (H2O2), and sodium hypochlorite, in both planktonic cells and submerged and air-liquid interface (ALI) biofilms. It also studied the roles of Pel and Psl in P. aeruginosa-Staphylococcus aureus interaction. The results demonstrate that the capacity of sublethal UVA exposure to increase cell hydrophobicity and cell attachment and generate cross-protection phenomena in P. aeruginosa depends on the presence of Pel and Psl. The study also shows that Pel and Psl have a key role in the tolerance to lethal doses of UVA radiation, sodium hypochlorite and H2O2, in both biofilms and planktonic cells. Finally, co-culture assays showed total inhibition of S. aureus growth in presence of P. aeruginosa. This phenomenon depends, at least in part, on the simultaneous presence of Pel and Psl in planktonic cells and biofilms, suggesting a relevant role of these polysaccharides in the interaction between these species.

Medidas de prevenção de lesão por pressão em enfermarias pediátricas: atuação dos profissionais de enfermagem / Pressure injury prevention measures in pediatric wards: nursing professionals' actions

RESUMO Objetivo classificar a qualidade da assistência de enfermagem e analisar a associação entre as caraterísticas dos profissionais e a execução de medidas preventivas de lesão por pressão em crianças hospitalizadas. Métodos estudo observacional, transversal, realizado com 235 profissionais de enfermagem. O instrumento de Prevenção de Lesão por Pressão possui três domínios, com 23 itens: Medidas preventivas e detecção precoce de lesão por pressão (9); Medidas de alívio de pressão (8) e Avaliação e notificação (6), analisados pelo Índice de Positividade para Qualidade da Assistência. Resultados predominou sexo feminino (98,7%), com idade média de 38,83 ± 9,94 anos, técnicos de enfermagem (57,4%) e com tempo de experiência profissional superior a cinco anos (77,1%). Constatou-se assistência predominantemente sofrível nos três domínios, em 82,6% das ações. Encontrou-se associação significante com as variáveis "participação em cursos" e "desgaste no trabalho" e uma tendência mais frequente de realização das medidas na faixa etária 31-40 anos. Conclusão a assistência de enfermagem foi predominantemente sofrível; a assistência associou-se à participação em cursos de aperfeiçoamento e desgaste no trabalho. Contribuições para a prática evidenciou-se a necessidade do investimento em capacitação profissional e oferta de insumos considerados indispensáveis para viabilizar uma assistência qualificada e segura.

ABSTRACT Objective to classify the quality of nursing care and analyze the association between professionals' characteristics and the implementation of preventive measures for pressure injuries in hospitalized children. Methods this observational, cross-sectional study involved 235 nursing professionals. The Pressure Injury Prevention instrument comprised three domains with a total of 23 items: Preventive measures and early detection of pressure injuries (9 items), Pressure relief measures (8 items), and Assessment and reporting (6 items), analyzed using the Positivity Index for Quality of Care. Results Most participants were females (98.7%), with a mean age of 38.83 ± 9.94 years, nursing technicians (57.4%), and had more than five years of experience (77.1%). Nursing care was predominantly inadequate across all three domains, with 82.6% of actions rated as poor. Significant associations were found between "participation in training courses" and "work-related exhaustion". There was a trend towards increased compliance with measurements among professionals aged 31-40. Conclusion nursing care was predominantly poor and participation in training courses and the presence of exhaustion were associated with better adherence to preventive measures. Contributions to practice the study highlights the pressing need for investments in professional training and the provision of necessary resources to support high-quality and safe nursing care.

UVA as environmental signal for alginate production in Pseudomonas aeruginosa: role of this polysaccharide in the protection of planktonic cells and biofilms against lethal UVA doses.

Pseudomonas aeruginosa is an extremely versatile microorganism that survives in a wide variety of niches. It is capable to respond rapidly to changes in the environment by producing secondary metabolites and virulence factors, including alginate. Alginate is an extracellular polysaccharide that protects the bacteria from antibiotics and oxidative agents, and enhances cell adhesion to solid surfaces in the process of biofilm formation. In the present study, we analyzed the role of alginate in the response of P. aeruginosa to lethal doses of ultraviolet-A (UVA) radiation, the major fraction of solar UV radiation reaching the Earth's surface. We also studied the role of alginate in the context of the adaptive responses generated when P. aeruginosa is exposed to sublethal doses of UVA radiation. The survival studies demonstrated that alginate has a key role in the resistance of P. aeruginosa to the oxidative stress generated by lethal UVA doses, both in planktonic cells and in static biofilms. In addition, the presence of alginate proved to be essential in the occurrence of adaptive responses such as induction of biofilm formation and cross-protection against hydrogen peroxide and sodium hypochlorite, both generated by exposure to low UVA doses. Finally, we demonstrated that the increase of biofilm formation is accompanied by an increase in alginate concentration in the biofilm matrix, possibly through the ppGpp-dependent induction of genes related to alginate regulation (algR and algU) and biosynthesis (algD operon). Given the importance of alginate in biofilm formation and its protective roles, better understanding of the mechanisms associated to its functions and synthesis is relevant, given the normal exposure of P. aeruginosa to UVA radiation and other types of oxidative stresses.

Homeophasic Adaptation in Response to UVA Radiation in Pseudomonas aeruginosa: Changes of Membrane Fatty Acid Composition and Induction of desA and desB Expression.

In bacteria, exposure to changes in environmental conditions can alter membrane fluidity, thereby affecting its essential functions in cell physiology. To adapt to these changes, bacteria maintain appropriate fluidity by varying the composition of the fatty acids of membrane phospholipids, a phenomenon known as homeophasic adaptation. In Pseudomonas aeruginosa, this response is achieved mainly by two mechanisms of fatty acid desaturation: the FabA-FabB and DesA-DesB systems. This study analyzed the effect of ultraviolet-A (UVA) radiation-the major fraction of solar UV radiation reaching the Earth's surface-on the homeophasic process. The prototypical strain PAO1 was grown under sublethal UVA doses or in the dark, and the profiles of membrane fatty acids were compared at early logarithmic, logarithmic and stationary growth phases. In the logarithmic growth phase, it was observed that growth under sublethal UVA doses induced the expression of the desaturase-encoding genes desA and desB and increased the proportion of unsaturated fatty acids; in addition, membrane fluidity could also increase, as suggested by the indices used as indicators of this parameter. The opposite effect was observed in the stationary growth phase. These results demonstrate the relevant role of UVA on the homeophasic response at transcriptional level.

Competências profissionais de promoção da saúde na prevenção de quedas na pediatria / Competencias profesionales de promoción de la salud en la prevención de caídas en pediatría / Professional health promotion competencies in preventing falls in pediatrics

Resumo Objetivo: Avaliar os domínios de competências de promoção da saúde dos profissionais, estabelecidos pelo Developing Competencies and Profissional Standards for Health Promotion Capacity Building in Europe, a partir da aplicação do instrumento de prevenção de quedas na pediatria. Métodos: Estudo transversal, descritivo, com abordagem quantitativa. Realizado nos meses de janeiro e fevereiro de 2018 nas unidades abertas de internação de dois hospitais de pediatria, com 203 profissionais de saúde (184 profissionais de enfermagem, dez médicos e nove fisioterapeutas), atuantes nas unidades abertas de internamento nas referidas instituições há pelo menos seis meses, a partir do autopreenchimento do formulário de caracterização profissional e do instrumento de prevenção de quedas na pediatria, o qual possui quatro fatores com 15 ações. A análise descritiva foi realizada de acordo com um checklist elaborado e validado que contém os fatores do instrumento de prevenção de quedas com as respectivas ações e os nove domínios de competências de promoção da saúde. Resultados: Os domínios de competência de promoção da saúde (Comunicação, Planejamento, Implementação, Avaliação e Pesquisa, Diagnóstico, Possibilidade de Mudanças e Parceria) foram identificados nas ações dos fatores do instrumento de prevenção de quedas na pediatria, em que 11 ações tiveram percentual de execução superior a 50%. Os domínios de competências Advocacia em saúde e Liderança não foram identificados. Conclusão: Identificaram-se sete domínios de competências de promoção da saúde, os quais são importantes para que se possa garantir cuidado seguro, integral e resolutivo para prevenção de quedas na pediatria.

Resumen Objetivo: Evaluar el dominio de las competencias de promoción de la salud de los profesionales, establecidos por el Developing Competencies and Profissional Standards for Health Promotion Capacity Building in Europe, a partir de la implementación del instrumento de prevención de caídas en pediatría. Métodos: Estudio transversal, descriptivo con enfoque cuantitativo. Llevado a cabo en los meses de enero y febrero de 2018 en las unidades abiertas de internación de dos hospitales de pediatría, con 203 profesionales de la salud (184 profesionales de enfermería, 10 médicos y 9 fisioterapeutas), que trabajan hace seis meses por lo menos en las unidades abiertas de internación en las instituciones mencionadas, mediante el autocompletado del formulario de caracterización profesional y el instrumento de prevención de caídas en pediatría, que posee 4 factores con 15 acciones. El análisis descriptivo fue realizado de acuerdo con una checklist elaborada y validada, que contiene los factores del instrumento de prevención de caídas con las respectivas acciones y los nueve dominios de competencias de promoción de la salud. Resultados: El dominio de las competencias de promoción de la salud (Comunicación, Planificación, Implementación, Evaluación e Investigación, Diagnóstico, Posibilidad de Cambios y Colaboración) fue identificado en las acciones de los factores del instrumento de prevención de caídas en pediatría, en que 11 acciones tuvieron un porcentaje de ejecución superior al 50 %. El dominio de las competencias Defensa en salud y Liderazgo no fue identificado. Conclusión: Se identificaron siete dominios de competencias de promoción de la salud, que son importantes para poder garantizar un cuidado seguro, integral y resolutivo para la prevención de caídas en pediatría.

Abstract Objective: To assess the domains of professional health promotion competencies, established by the Developing Competencies and Professional Standards for Health Promotion Capacity Building in Europe, based on applying the Instrument of Prevention of Falls in Pediatrics. Methods: This is a cross-sectional, descriptive and quantitative study. It was carried out in January and February 2018 in open inpatient units of two pediatric hospitals with 203 health professionals (184 nursing professionals, ten doctors and nine physiotherapists); working in open inpatient units at the referred institutions for at least six months; from self-completion of the professional characterization form and the Instrument of Prevention of Falls in Pediatrics, which has four factors with 15 actions. Descriptive analysis was performed according to a checklist developed and validated that contains the factors of the instrument for preventing falls with the respective actions and the nine domains of health promotion competency. Results: The health promotion competency domains (Communication, Planning, Implementation, Assessment, Evaluation and Research, Enable Change, and Mediate through Partnership) were identified in the actions of the factors of the Instrument of Prevention of Falls in Pediatrics. Eleven actions had an execution percentage higher than 50%. The Advocate for Health and Leadership domains have not been identified. Conclusion: Seven domains of health promotion competencies were identified, which are important to ensure safe, comprehensive and resolutive care for preventing falls in pediatrics.

Prescrições e aprazamentos de medicamentos endovenosos em pediatria: estudo descritivo / Prescripciones y horarios de fármacos endovenosos en pediatría: estudio descriptivo / Prescriptions and scheduling of endovenous medications in pediatrics: descriptive study

RESUMO Objetivos: analisar as prescrições e o aprazamento de medicamentos endovenosos a crianças e adolescentes hospitalizados, de acordo com as recomendações do Protocolo de Segurança na Prescrição, Uso e Administração de Medicamentos. Métodos: estudo descritivo, documental, com análise de 352 prescrições de medicamentos endovenosos, em unidades de internação pediátrica. A faixa etária das crianças era de 29 dias até 16 anos de idade completos. Coleta dos dados ocorreu de agosto a novembro de 2017. Utilizou-se instrumento do tipo checklist. A análise foi realizada por meio da estatística descritiva, obedecendo aos preceitos éticos. Resultados: constatou-se que o desempenho foi satisfatório em relação às prescrições e estavam de acordo com o recomendado pelo Ministério da Saúde do Brasil. Os itens essenciais nas prescrições medicamentosas apresentaram as seguintes incidências: data (98%), nome genérico (95,2%), concentração (98,6%), dose (99,7%), via de administração (95,7%), posologia (98,9%), orientações (87,5%), assinatura do médico (99,1%), carimbo médico (97,7%), assinatura do enfermeiro (93,2%) e carimbo do enfermeiro (84,7%). Nas 352 prescrições, analisaram-se 1.069 medicamentos, dos quais 1.059 (99,06%) apresentaram dados satisfatórios quanto à checagem dos medicamentos endovenosos. Conclusões: a maioria dos itens da prescrição foi satisfatória, contudo, alguns itens foram considerados insatisfatórios, tendo ações não realizadas corretamente.

RESUMEN Objetivos: analizar la prescripción y programación de medicamentos endovenosos para niños y adolescentes hospitalizados, de acuerdo con las recomendaciones del Protocolo de Seguridad en la Prescripción, Uso y Administración de Medicamentos. Métodos: estudio descriptivo, documental, con análisis de 352 prescripciones de medicamentos endovenosos en unidades de hospitalización pediátrica. El rango de edad de los niños fue de 29 días a 16 años. La recolección de datos se llevó a cabo de agosto a noviembre de 2017. Se utilizó un instrumento tipo lista de verificación. El análisis se realizó mediante estadística descriptiva, cumpliendo con los preceptos éticos. Resultados: se encontró que el desempeño fue satisfactorio con relación a las prescripciones y estuvo de acuerdo con lo recomendado por el Ministerio de Salud de Brasil. Los ítems esenciales en la prescripción de medicamentos tuvieron las siguientes incidencias: fecha (98%), nombre genérico (95,2%), concentración (98,6%), dosis (99,7%), vía de administración (95,7%), posología (98,9%), directrices (87,5%), firma del médico (99,1%), sello médico (97,7%), firma de la enfermera (93,2%) y sello de la enfermera (84,7%). En las 352 prescripciones se analizaron 1.069 medicamentos, de los cuales 1.059 (99,06%) presentaron datos satisfactorios en cuanto a la verificación de medicamentos endovenosos. Conclusiones: la mayoría de los ítems de prescripción fueron satisfactorios, sin embargo, algunos ítems se consideraron insatisfactorios, con acciones no realizadas correctamente.

ABSTRACT Objectives: to analyze the prescriptions and scheduling of intravenous medications for hospitalized children and adolescents, following the recommendations of the Safety Protocol in the Prescription, Use, and Administration of Medicines. Methods: descriptive, documentary study, with analysis of 352 prescriptions for intravenous medications in pediatric inpatient units. The age range of the children was 29 days up to 16 years old. Data collection took place from August to November 2017. A checklist-type instrument was used. The analysis was performed using descriptive statistics, in compliance with ethical precepts. Results: we found that the performance was satisfactory concerning the prescriptions and followed the recommendations of the Ministry of Health of Brazil. The essential items in medication prescriptions had the following incidences: date (98%), generic name (95.2%), concentration (98.6%), dose (99.7%), route of administration (95.7% ), dosage (98.9%), guidelines (87.5%), physician's signature (99.1%), medical stamp (97.7%), nurse's signature (93.2%) and nurse's stamp (84.7%). In the 352 prescriptions, 1,069 medications were analyzed, of which 1,059 (99.06%) presented satisfactory data regarding the verification of intravenous medications. Conclusions: most of the prescription items were satisfactory; however, some items were considered unsatisfactory, with actions not performed correctly.

Effect of temperature on microcystin-LR removal and lysis activity on Microcystis aeruginosa (cyanobacteria) by an indigenous bacterium belonging to the genus Achromobacter.

Microcystis is a frequent cyanobacterium bloom-forming with cosmopolitan distribution which can produce a hepatotoxin group called microcystins (MCs). These MCs are resistant to the traditional processes employed in the water treatment plants and they are often detected after conventional treatments. Because of this, the bio-removal studies have obtained a great interest in the last decades. In this work, a bacterial strain namely LG1 with the ability to remove microcystin-LR (MC-LR) under laboratory conditions was isolated from Rio de la Plata River and it was identified as Achromobacter spp. This ubiquitous bacterium was able to remove 79.5% MC-LR in 7 days with average removal time of 3.33 ± 0.08, 3.06 ± 0.05, and 2.77 ± 0.05 days at 28, 32, and 36 ± 1 °C, being higher at high temperature (36 °C) with an activation energy = 16.79 ± 1.99 kJ mol-1. LG1 grew better at higher temperature (from 28 to 36 ± 1 °C) increasing the specific growth rate (µ) and reducing 2-fold the lag phase duration (LPD) without significant differences (p > 0.05) between maximum population density (MPD). In addition, LG1 showed a lysis activity on two M. aeruginosa native strains in 7 days measured as chlorophyll a (Chl-a) concentration. The lysis activity increased around 2-fold when increasing the temperature from 28 to 36 ± 1 °C. This is the first report of an indigenous bacterium belonging to the genus Achromobacter spp. isolated from the Rio de la Plata River with the capacity to remove MC-LR and lysis activity on M. aeruginosa.

Role of quorum sensing in UVA-induced biofilm formation in Pseudomonas aeruginosa.

Pseudomonas aeruginosa, a versatile bacterium present in terrestrial and aquatic environments and a relevant opportunistic human pathogen, is largely known for the production of robust biofilms. The unique properties of these structures complicate biofilm eradication, because they make the biofilms very resistant to diverse antibacterial agents. Biofilm development and establishment is a complex process regulated by multiple regulatory genetic systems, among them is quorum sensing (QS), a mechanism employed by bacteria to regulate gene transcription in response to population density. In addition, environmental factors such as UVA radiation (400-315 nm) have been linked to biofilm formation. In this work, we further investigate the mechanism underlying the induction of biofilm formation by UVA, analysing the role of QS in this phenomenon. We demonstrate that UVA induces key genes of the Las and Rhl QS systems at the transcriptional level. We also report that pelA and pslA genes, which are essential for biofilm formation and whose transcription depends in part on QS, are significantly induced under UVA exposure. Finally, the results demonstrate that in a relA strain (impaired for ppGpp production), the UVA treatment does not induce biofilm formation or QS genes, suggesting that the increase of biofilm formation due to exposure to UVA in P. aeruginosa could rely on a ppGpp-dependent QS induction.

Two Functional Variants of AP-1 Complexes Composed of either γ2 or γ1 Subunits Are Independently Required for Major Histocompatibility Complex Class I Downregulation by HIV-1 Nef.

The HIV-1 accessory protein Nef downregulates the cell surface expression of major histocompatibility complex class I (MHC-I) molecules to facilitate virus spreading. The Nef-induced downregulation of MHC-I molecules such as HLA-A requires the clathrin adaptor protein 1 (AP-1) complex. The cooperative interaction of Nef, AP-1, and the cytosolic tail (CT) of HLA-A leads to a redirection of HLA-A targeting from the trans-Golgi network (TGN) to lysosomes for degradation. Although the γ-adaptin subunit of AP-1 has two distinct isoforms (γ1 and γ2), which may form two AP-1 complex variants, so far, only the importance of AP-1γ1 in MHC-I downregulation by Nef has been investigated. Here, we report that the AP-1γ2 isoform also participates in this process. We found that AP-1γ2 forms a complex with Nef and HLA-A2_CT and that this interaction depends on the Y320 residue in HLA-A2_CT and Nef expression. Moreover, Nef targets AP-1γ1 and AP-1γ2 to different compartments in T cells, and the depletion of either AP-1 variant impairs the Nef-mediated reduction of total endogenous HLA-A levels and rescues HLA-A levels on the cell surface. Finally, immunofluorescence and immunoelectron microscopy analyses reveal that the depletion of γ2 in T cells compromises both the Nef-mediated retention of HLA-A molecules in the TGN and targeting to multivesicular bodies/late endosomes. Altogether, these results show that in addition to AP-1γ1, Nef also requires the AP-1γ2 variant for efficient MHC-I downregulation.IMPORTANCE HIV-1 Nef mediates evasion of the host immune system by inhibiting MHC-I surface presentation of viral antigens. To achieve this goal, Nef modifies the intracellular trafficking of MHC-I molecules in several ways. Despite being the subject of intense study, the molecular details underlying these modifications are not yet fully understood. Adaptor protein 1 (AP-1) plays an essential role in the Nef-mediated downregulation of MHC-I molecules such as HLA-A in different cell types. However, AP-1 has two functionally distinct variants composed of either γ1 or γ2 subunit isoforms. Because previous studies on the role of AP-1 in MHC-I downregulation by Nef focused on AP-1γ1, an important open question is the participation of AP-1γ2 in this process. Here, we show that AP-1γ2 is also essential for Nef-mediated depletion of surface HLA-A molecules in T cells. Our results indicate that Nef hijacks AP-1γ2 to modify HLA-A intracellular transport, redirecting these proteins to lysosomes for degradation.

Evaluation of Viable Cells in Pseudomonas aeruginosa Biofilmsby Colony Count and Live/Dead Staining.

Pseudomonas aeruginosa is a human pathogen capable to form robust biofilms. P. aeruginosa biofilms represent a serious problem because of the adverse effects on human health and industry, from sanitary and economic points of view. Typical strategies to break down biofilms have been long used, such as the use of disinfectants or antibiotics, but also, according to their high resistance to standard antimicrobial approaches, alternative strategies employing photocatalysis or control of biofilm formation by modifying surfaces, have been proposed. Colony forming units (cfu) counting and live/dead staining, two classic techniques used for biofilm quantification, are detailed in this work. Both methods assess cell viability, a key factor to analyze the microbial susceptibility to given treatment, then, they represent a good approach for evaluation of an antibiofilm strategy.

Response to lethal UVA radiation in the Antarctic bacterium Pseudomonas extremaustralis: polyhydroxybutyrate and cold adaptation as protective factors.

Pseudomonas extremaustralis is an Antarctic bacterium with high stress resistance, able to grow under cold conditions. It is capable to produce polyhydroxyalkanoates (PHAs) mainly as polyhydroxybutyrate (PHB) and, to a lesser extent, medium-chain length polyhydroxyalkanoates (mclPHAs). In this work, we analyzed the role of PHAs and cold adaptation in the survival of P. extremaustralis after lethal UVA exposure. P. extremaustralis presented higher radiation resistance under polymer accumulation conditions. This result was also observed in the derivative mutant strain PHA-, deficient for mclPHAs production. On the contrary, the PHB- derivative mutant, deficient for PHB production, showed high sensitivity to UVA exposure. Complementation of the PHB- strain restored the wild-type resistance level, indicating that the UVA-sensitive phenotype is due to the lack of PHB. All strains exhibited high sensitivity to radiation when cultured under PHAs non-accumulation conditions. A slight decrease in PHB content was observed after UVA exposure in association with increased survival. The scattering of UVA radiation by intracellular PHAs granules could also result in bacterial cell protection. In addition, cold conditions improved UVA tolerance, probably depending on PHB mobilization. Results showed that PHB accumulation is crucial in the resistance to UVA in P. extremaustralis. Mechanisms involved probably entail depolymerization and light scattering acting as a screen, both conferring protection against oxidative stress.

Segurança do paciente pediátrico no processo de administração de medicamento endovenoso / Pediatric patient safety in the intravenous drug administration process

Objetivo: Avaliar o processo de preparo e administração de medicamentos endovenosos em um hospital pediátrico. Métodos: Estudo transversal de natureza quantitativa. Foram realizadas 135 observações do processo de preparo e administração de medicamentos por via endovenosa em crianças. Para a avaliação do desempenho dos profissionais na execução de cada ação do processo determinou-se o Índice de Positividade para Qualidade da Assistência. A análise foi através de estatística descritiva obedecendo aos preceitos éticos. Resultados: Na etapa de leitura de prescrição médica uma ação obteve desempenho satisfatório (igual ou maior que 70%). As ações de todas as 15 ações da etapa de organização do ambiente e seleção dos materiais nove foram satisfatórias (maior que 70%). Conclusão: Os achados referentes ao estudo nos permite evidenciar que existem falhas no processo de preparo e administração de medicamentos. É imprescindível a melhoria dos cuidados em saúde através de educação permanente. (AU)

Objective: To evaluate the process of preparation and administration of intravenous drugs in a pediatric hospital. Methods: Cross-sectional study of a quantitative nature. 135 observations were made of the process of preparation and administration of drugs intravenously in children. To assess the performance of professionals in the execution of each action in the process, the Positivity Index for Quality of Care was determined. The analysis was through descriptive statistics obeying the ethical precepts. Results: In the step prior to the preparation of the medications, all actions achieved satisfactory performance (≥70%). Of the fifteen actions for preparing medications, nine were satisfactory (≥70%). During and after the administration of six of the seven actions, they obtained satisfactory performance (≥70%). Conclusion: The findings regarding the study allow us to evidence that there are flaws in the process of preparing and administering medications. (AU)

Objetivo: evaluar el proceso de preparación y administración de drogas intravenosas en un hospital pediátrico. Métodos: estudio transversal de naturaleza cuantitativa. Se hicieron 135 observaciones sobre el proceso de preparación y administración de drogas por vía intravenosa en niños. Para evaluar el desempeño de los profesionales en la ejecución de cada acción en el proceso, se determinó el Índice de Positividad para la Calidad de la Atención. El análisis fue a través de estadísticas descriptivas que obedecen los preceptos éticos. Resultados: En el paso previo a la preparación de los medicamentos, todas las acciones lograron un desempeño satisfactorio (≥70%). De las quince acciones para preparar medicamentos, nueve fueron satisfactorias (≥70%). Durante y después de la administración de seis de las siete acciones, obtuvieron un desempeño satisfactorio (≥70%). Conclusión: Los hallazgos con respecto al estudio nos permiten evidenciar que hay fallas en el proceso de preparación y administración de medicamentos. (AU)

Assessment of rapid direct E-test on positive blood culture for same-day antimicrobial susceptibility.

INTRODUCTION: Early and appropriated antimicrobial therapy showed to positively impact on the clinical improvement of septic patients. The aim of this study was to evaluate E-test methodology to obtain rapid results of antimicrobial susceptibility, starting directly from blood culture bottles positive to Gram-negative monomicrobial flora. MATERIALS AND METHODS: One hundred and five blood culture samples positive to Gram-negative rods at the microscopic examination were collected. Bacterial identification from early subculture on blood agar after 4 h incubation and rapid direct E-test from blood culture broth were performed on every sample. Antibiotics MIC were achieved after 5-6 h of incubation. Resulting MIC values were compared with those obtained with reference E-test from the overnight subculture. Categorical agreement (CA) and essential agreement (EA) were evaluated. RESULTS: Comparison between rapid direct E-test and reference E-test showed CA ranging from 95.1 to 100 % and 88.2 to 100 % for Enterobacteriaceae (EB) and for non-fermenting Gram-negative bacilli, respectively. Rapid direct E-test showed an overall EA of 80.1 %, revealing different EA rates for the tested antibiotics. Among carbapenemase-producing EB, CA of 87.5 % and EA of 75.5 % for MP were achieved. DISCUSSION: The same-day communication of the antimicrobial susceptibility represents an important challenge in the multidrug-resistance era. Despite not being able to anticipate actual MIC values, the rapid direct E-test may be useful to obtain preliminary AST results in 5-6 h, especially if used in association with phenotypic or genotypic tests to identify the main resistance mechanisms.

The Ambiguous Roles of Extracellular Vesicles in HIV Replication and Pathogenesis.

Cells from all kingdoms of life can release membrane-enclosed vesicles to the extracellular milieu. These extracellular vesicles (EVs) may function as mediators of intercellular communication, allowing the transfer of biologically active molecules between cells and organisms. It has become clear that HIV particles and certain types of EVs, such as exosomes, share many similarities regarding morphology, composition, and biogenesis. This review presents a summary of the literature describing the intricate relationship between HIV and EVs biogenesis. Also, we discuss the latest progress toward understanding the mechanisms by which EVs influence HIV pathogenesis, as well as, how HIV modulates EVs composition in infected cells to facilitate viral spread.

Exposure to low doses of UVA increases biofilm formation in Pseudomonas aeruginosa.

The establishment of bacterial biofilms on abiotic surfaces is a complex process regulated by multiple genetic regulators and environmental factors which are able to modulate the passage of planktonic cells to a sessile state. Solar ultraviolet-A radiation (UVA, 315-400) is one of the main environmental stress factors that bacteria must face at the Earth´s surface. The deleterious effects of UVA are mainly due to oxidative damage. This paper reports that exposure to low UVA doses promotes biofilm formation in three prototypical strains of Pseudomonas aeruginosa, a relevant opportunistic human pathogen. It demonstrates that exposure of planktonic cells to sublethal doses of UVA can increase cell surface hydrophobicity and swimming motility, two parameters known to favor cell adhesion. These results suggest that UVA radiation acts, at least in part, by promoting the first stages of biofilm development.

New envelope stress factors involved in σE activation and conditional lethality of rpoE mutations in Salmonella enterica.

Salmonella enterica serovar Typhimurium (S. typhimurium) can cause food- and water-borne illness with diverse clinical manifestations. One key factor for S. typhimurium pathogenesis is the alternative sigma factor σE, which is encoded by the rpoE gene and controls the transcription of genes required for outer-membrane integrity in response to alterations in the bacterial envelope. The canonical pathway for σE activation involves proteolysis of the antisigma factor RseA, which is triggered by unfolded outer-membrane porins (OMPs) and lipopolysaccharides (LPS) that have accumulated in the periplasm. This study reports new stress factors that are able to activate σE expression. We demonstrate that UVA radiation induces σE activity in a pathway that is dependent on the stringent response regulator ppGpp. Survival assays revealed that rpoE has a role in the defence against lethal UVA doses that is mediated by functions that are dependent on and independent of the alternative sigma factor RpoS. We also report that the envelope stress generated by phage infection requires a functional rpoE gene for optimal bacterial tolerance and that it is able to induce σE activity in an RseA-dependent fashion. σE activity is also induced by hypo-osmotic shock in the absence of osmoregulated periplasmic glucans (OPGs). It is known that the rpoE gene is not essential in S. typhimurium. However, we report here two cases of the conditional lethality of rpoE mutations in this micro-organism. We demonstrate that rpoE mutations are not tolerated in the absence of OPGs (at low to moderate osmolarity) or LPS O-antigen. The latter case resembles that of the prototypic Escherichia coli strain K12, which neither synthesizes a complete LPS nor tolerates null rpoE mutations.