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Bacterial coaggregation is a highly specific type of cell-cell interaction, well-documented among oral bacteria, and involves specific characteristics of the cell surface of the coaggregating strains. However, the understanding of the mechanisms promoting coaggregation in aquatic systems remains limited. This gap is critical to address, given the broad implications of coaggregation for multispecies biofilm formation, water quality, the performance of engineered systems, and diverse biotechnological applications. Therefore, this study aims to comprehensively characterize the cell surface of the coaggregating strain Delftia acidovorans 005P, isolated from drinking water, alongside a non-coaggregating strain, D. acidovorans 009P. By analyzing two strains of the same species, we aim to identify the factors contributing to the coaggregation ability of strain 005P. To achieve this, we employed a combination of physicochemical characterization, Fourier-transform infrared spectroscopy (FTIR), and advancing imaging techniques [transmission electron microscopy and cryo-electron tomography (cryo-ET)]. The coaggregating strain (005P) exhibited higher surface hydrophobicity, negative surface charge, and cell surface and co-adhesion energies than the non-coaggregating strain (009P). The chemical characterization of bacterial surfaces through FTIR revealed subtle differences, particularly in spectral regions linked to carbohydrates and phosphodiesters/amide III of proteins (860-930 cm -1 and 1212-1240 cm -1 , respectively). Cryo-ET highlighted significant differences in pili structures between the strains, such as variations in length, frequency, and arrangement. The pili in the 005P strain, identified as pili-like adhesins, serve as key mediators of coaggregation. By integrating physicochemical analyses and high-resolution imaging techniques, this study conclusively links the coaggregation ability of D. acidovorans 005P to its unique pili characteristics, emphasizing their crucial role in microbial coaggregation in aquatic environments., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

This work aimed to explore an alternative to the use of antibiotics for prevention and treatment of wounds infection caused by two common bacterial pathogens Staphylococcus aureus and Pseudomonas aeruginosa. For this purpose, three different essential oil components (EOCs), namely carvacrol, citronellol and cinnamic acid, were loaded into electrospun fibers of poly-ε-caprolactone (PCL) aided by alpha-cyclodextrin (αCD) and hydroxypropyl-β-cyclodextrin (HPβCD). Electrospun-fibers prepared with each EOC and their mixtures were screened for antimicrobial capability and characterized regarding morphological, mechanical, thermal, surface polarity, antibiofilm and antioxidant properties. αCD formed poly(pseudo)rotaxanes with PCL and weakly interacted with EOCs, while HPβCD facilitated EOC encapsulation and formation of homogeneous fibers (500-1000 nm diameter) without beads. PCL/HPβCD fibers with high concentration of EOCs (mainly carvacrol and cinnamic acid) showed strong antibiofilm (>3 log CFU reduction) and antioxidant activity (10-50% DPPH scavenging effects). Different performances were recorded for the EOCs and their mixtures; cinnamic acid migrated to fiber surface and was released faster. Fibers biocompatibility was verified using hemolysis tests and in ovo tissue integration and angiogenesis assays. Overall, HPβCD facilitates complete release of EOCs from the fibers to the aqueous medium, being an environment-friendly and cost-effective strategy for the treatment of infected wounds., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Biofilms in drinking water (DW) systems persistently challenge traditional disinfection methods due to intricate microbial interactions, with coaggregation playing a crucial role in forming multispecies biofilms. This study examined the implications of coaggregation on tolerance towards sodium hypochlorite (NaOCl) disinfection. Dual-species biofilms were formed for seven days on polyvinyl chloride coupons, comprising a strain of the emerging pathogen Stenotrophomonas maltophilia and the coaggregating strain Delftia acidovorans 005 P. For comparison, dual-species biofilms were also formed with a non-coaggregation strain (D. acidovorans 009 P). The minimum bactericidal concentration (MBC) for each planktonic strain varied (D. acidovorans: 1 mg/L, S. maltophilia: 1.5 mg/L) below the safe DW treatment limits. However, high NaOCl doses (10 ×MBC and 100 ×MBC,) showed low efficacy against dual-species biofilms, indicating significant biofilm tolerance to disinfection. Membrane damage occurred at sub-MBC without culturability loss, underscoring biofilm resilience. The biofilm analysis revealed a complex interplay between the composition of extracellular polymeric substances and the architecture, which was influenced by the presence of the coaggregating strain. Overall, coaggregation significantly influenced biofilm formation and resilience, impacting NaOCl disinfection. These findings underscore the challenges of microbial interactions in biofilms, emphasizing the need for improved disinfection strategies to control biofilms in drinking water systems., Competing Interests: Declaration of Competing Interest 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 paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The biofilm architecture is significantly influenced by external environmental conditions. Biofilms grown on drinking water distribution systems (DWDS) are exposed to environmental contaminants, including parabens, and disinfection strategies, such as chlorine. Although changes in biofilm density and culturability from chemical exposure are widely reported, little is known about the effects of parabens and chlorine on biofilm morphology and architecture. This is the first study evaluating architectural changes in Stenotrophomonas maltophilia colony biofilms (representatives of bacterial communities presented in DWDS) induced by the exposure to methylparaben (MP) at environmental (15 μg/L) and in-use (15 mg/L) concentrations, and chlorine at 5 mg/L, using widefield epi-fluorescence mesoscopy with Mesolens. The GFP fluorescence of colony biofilms allowed the visualization of internal structures and Nile Red fluorescence permitted the inspection of the distribution of lipids. Our data show that exposure to MP triggers physiological and morphological adaptation in mature colony biofilms by increasing the complexity of internal structures, which may confer protection to embedded cells from external chemical molecules. These architectural modifications include changes in lipid distribution as an adaptive response to MP exposure. Although chlorine exposure affected colony biofilm diameter and architecture, the colony roundness was completely affected by the simultaneous presence of MP and chlorine. This work is pioneer in using Mesolens to highlight the risks of exposure to emerging environmental contaminants (MP), by affecting the architecture of biofilms formed by drinking water (DW) bacteria, even when combined with routine disinfection strategies., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2024. Published by Elsevier B.V.)

Several studies have linked air pollution to COVID-19 morbidity and severity. However, these studies do not account for exposure levels to SARS-CoV-2, nor for different sources of air pollution. We analyzed individual-level data for 8.3 million adults in the Netherlands to assess associations between long-term exposure to ambient air pollution and SARS-CoV-2 infection (i.e., positive test) and COVID-19 hospitalisation risks, accounting for spatiotemporal variation in SARS-CoV-2 exposure levels during the first two major epidemic waves (February 2020-February 2021). We estimated average annual concentrations of PM 10 , PM 2.5 and NO 2 at residential addresses, overall and by PM source (road traffic, industry, livestock, other agricultural sources, foreign sources, other Dutch sources), at 1 × 1 km resolution, and weekly SARS-CoV-2 exposure at municipal level. Using generalized additive models, we performed interval-censored survival analyses to assess associations between individuals' average exposure to PM 10 , PM 2.5 and NO 2 in the three years before the pandemic (2017-2019) and COVID-19-outcomes, adjusting for SARS-CoV-2 exposure, individual and area-specific confounders. In single-pollutant models, per interquartile (IQR) increase in exposure, PM 10 was associated with 7% increased infection risk and 16% increased hospitalisation risk, PM 2.5 with 8% increased infection risk and 18% increased hospitalisation risk, and NO 2 with 3% increased infection risk and 11% increased hospitalisation risk. Bi-pollutant models suggested that effects were mainly driven by PM. Associations for PM were confirmed when stratifying by urbanization degree, epidemic wave and testing policy. All emission sources of PM, except industry, showed adverse effects on both outcomes. Livestock showed the most detrimental effects per unit exposure, whereas road traffic affected severity (hospitalisation) more than infection risk. This study shows that long-term exposure to air pollution increases both SARS-CoV-2 infection and COVID-19 hospitalisation risks, even after controlling for SARS-CoV-2 exposure levels, and that PM may have differential effects on these COVID-19 outcomes depending on the emission source., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Professionals in the justice system are particularly susceptible to occupational stress and burnout due to factors intrinsic to their profession. The Forensic Professional's Stress Inventory (FPSI) was designed to assess stress and psychological distress specifically in justice system professionals. A preliminary 41-item scale was administered to a sample of 690 forensic professionals (i.e., judges, lawyers, and attorneys). Exploratory factor analysis, exploratory structural equation modeling, and confirmatory factor analysis were conducted to find the most interpretable and parsimonious factor solution for FPSI. The 25-item bifactor model (with four first-order factors) demonstrated the most adequate fit to the data. Overall, FPSI revealed adequate psychometric properties and would be a useful instrument for assessing psychological strain and stress in forensic professionals., Competing Interests: Declaration of competing interest The authors declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The increase of bacterial resistance to antibiotics is a growing concern worldwide and the search for new therapies could cost billions of dollars and countless lives. Inert surfaces are major sources of contamination due to easier adhesion and formation of bacterial biofilms, hindering the disinfection process. Therefore, the objective of this study was to develop a photoactivatable and anti-adhesive kappa-carrageenan coating using proanthocyanidin as a photosensitizer. The complete reduction (>5-log 10 CFU/cm 3 ) of culturable cells of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa pathogens was achieved after 30 min of exposure to visible light (420 nm; 30 mW/cm 2 ) with 5 % (w/v) of the photosensitizer. Cell membrane damage was confirmed by measuring potassium leakage, epifluorescence microscopy and bacterial motility analysis. Overall, visible light irradiation on coated solid surfaces mediated by proanthocyanidin showed no cytotoxicity and inactivated clinically important pathogens through the generation of reactive oxygen species, inhibiting bacterial initial adhesion. The developed coating is a promising alternative for a wide range of applications related to surface disinfection and food biopreservation., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The contamination of indoor areas is a global health problem that can cause the dispersion of infectious diseases. In that sense, it is urgent to find new strategies applying a lower concentration of the traditional chemicals used for cleaning and disinfection. Ultraviolet radiation (UV), in particular far-UV-C (200-225 nm), has emerged as a successful, powerful, easy-to-apply, and inexpensive approach for bacterial eradication that still requires scientific assessment. This study investigated new strategies for disinfection based on far-UV-C (222 nm) combined with chlorine and mechanical cleaning, providing an innovative solution using low doses. The bactericidal activity of far-UV-C (222 nm) was tested at an intensity of irradiation from 78.4 μW/cm 2 to 597.7 μW/cm 2 (for 1 min) against Escherichia coli and Staphylococcus epidermidis adhered on polystyrene microtiter plates. It was further tested in combination with mechanical cleaning (ultrasounds for 1 min) and free chlorine (0.1, 0.5, and 1 mg/L for 5 min). The triple combination consisting of mechanical cleaning + free chlorine (0.5 mg/L) + far-UV-C (54 mJ/cm 2 ) was tested against cells adhered to materials found in hospital settings and other public spaces: polyvinyl chloride (PVC), stainless steel (SS), and polyetheretherketone (PEEK). Disinfection with far-UV-C (54 mJ/cm 2 ) and free chlorine at 0.5 mg/L for 5 min allowed a total reduction of culturable E. coli cells and a logarithmic reduction of 2.98 ± 0.03 for S. epidermidis. The triple combination of far-UV-C, free chlorine, and mechanical cleaning resulted in a total reduction of culturable cells for both adhered bacteria. Bacterial adhesion to PVC, SS, and PEEK occurred at distinct extents and influenced the bactericidal activity of the triple combination, with logarithmic reductions of up to three. The overall results highlight that, based on culturability assessment, far-UV-C (54 mJ/cm 2 ) with chlorine (0.5 mg/L; 5 min) and mechanical cleaning (1 min) as an efficient disinfection strategy using mild conditions. The combination of culturability and viability assessment of disinfection is recommended to detect regrowth events and increase the effectiveness in microbial growth control., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or conflicts of interest in the writing of this article., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Among different pollutants of emerging concern, parabens have gained rising interest due to their widespread detection in water sources worldwide. This occurs because parabens are used in personal care products, pharmaceuticals, and food, in which residues are generated and released into aquatic environments. The regulation of the use of parabens varies across different geographic regions, resulting in diverse concentrations observed globally. Concentrations of parabens exceeding 100 μg/L have been found in wastewater treatment plants and surface waters while drinking water (DW) sources typically exhibit concentrations below 6 μg/L. Despite their low levels, the presence of parabens in DW is a potential exposure route for humans, raising concerns for both human health and environmental microbiota. Although a few studies have reported alterations in the functions and characteristics of microbial communities following exposure to emerging contaminants, the impact of the exposure to parabens by microbial communities, particularly biofilm colonizers, remains largely understudied. This review gathers the most recent information on the occurrence of parabens in water sources, as well as their effects on human health and aquatic organisms. The interactions of parabens with microbial communities are reviewed for the first time, filling the knowledge gaps on the effects of paraben exposure on microbial ecosystems and their impact on disinfection tolerance and antimicrobial resistance, with potential implications for public health., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Although it is considered the reference for quantification of neutralizing antibodies, classical method of the plaque reduction neutralization test (PRNT) is labor intensive, requires specific equipment and inputs, besides a long time for its finalization, even in the micro-PRNT version (in 96-well plates). It has a higher sample throughput, however the smaller wells make the reading of plaques more difficult. With an immunoenzymatic revelation step and a semi-automated reading, the μFRN-HRP (micro Focus Reduction Neutralization - Horseradish Peroxidase) is a faster and more efficient test for the quantification of YF neutralizing antibodies. This study aimed to standardize, validate, and compare it with the reference method in 6-well plates (PRNT). Once the execution protocol was standardized, precision, accuracy, selectivity, and robustness were evaluated to validate the μFRN-HRP. In addition, 200 sera of vaccinees were processed by the μFRN-HRP and by the micro-PRNT to compare with the reference test, estimating agreement by Intraclass Correlation Coefficient (ICC). The standardization and validation of the μFRN-HRP was carried out successfully. Weak to moderate agreement was observed between μFRN-HRP and PRNT for titers in reciprocal dilution, while the same comparison between the classical tests resulted in a better ICC. However, titers in milli-international units obtained by μFRN-HRP showed a substantial agreement with PRNT, while the agreement between micro-PRNT and PRNT was inferior. Therefore, μFRN-HRP can be used in the confirmation of natural YF infection and immune response to vaccination, replacing the micro-PRNT, gaining agility, while preserving the specificity of the result., (Copyright © 2023. Published by Elsevier B.V.)

The potential of pentapotassium bis(peroxymonosulphate) bis(sulphate) (OXONE) to control biofilms in drinking water distribution systems (DWDS) was evaluated and compared to chlorine disinfection. Mature biofilms of drinking water (DW)-isolated Stenotrophomonas maltophilia were formed using a simulated DWDS with a rotating cylinder reactor (RCR). After 30 min of exposure, OXONE at 10 × minimum bactericidal concentration (MBC) caused a significant 4 log reduction of biofilm culturability in comparison to the unexposed biofilms and a decrease in the number of non-damaged cells below the detection limit (4.8 log cells/cm 2 ). The effects of free chlorine were restricted to approximately 1 log reduction in both biofilm culturability and non-damaged cells. OXONE in synthetic tap water (STW) at 25 ºC was more stable over 40 days than free chlorine in the same conditions. OXONE solution exhibited a disinfectant decrease of about 10% of the initial concentration during the first 9 days, and after this time the values remained stable. Whereas possible reaction of chlorine with inorganic and organic substances in STW contributed to free chlorine depletion of approximately 48% of the initial concentration. Electron paramagnetic resonance (EPR) spectroscopy studies confirmed the presence of singlet oxygen and other free radicals during S. maltophilia disinfection with OXONE. Overall, OXONE constitutes a relevant alternative to conventional DW disinfection for effective biofilm control in DWDS., Competing Interests: Declaration of Competing Interest 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 paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: To describe trends in the prevalence of diabetes mellitus (DM) in Brazil and to analyze its association with the consumption of artificially sweetened beverages among individuals aged 18 years or older., Study Design: This was a repeated cross-sectional study., Methods: Annual data from VIGITEL surveys (2006-2020) were used, which included adults from all Brazilian state capitals. The outcome was the prevalence of DM (type 1 and type 2). The main exposure variable was consuming beverages like soft drinks and artificial juices, either in its 'diet, light, or zero' form. Covariates included sex, age, sociodemographic characteristics, smoking, alcohol consumption, physical activity, fruit consumption, and obesity. The temporal trend in the indicators and the etiological fraction (population attributable risk [PAR]) were calculated. Analyses were performed using Poisson regression. The association between DM and consumption of beverages was tested, excluding the year 2020 due to the pandemic; restricting the analysis to the final three years (2018-2020)., Results: Overall, 757,386 subjects were included. The prevalence of DM increased from 5.5% to 8.2%, with an annual growth of 0.17 percentage points (95% CI 0.11-0.24). Among those who consumed diet/light/zero beverages, the annual percentage change of DM was four times greater. The PAR corresponding to the consumption of diet/light/zero beverages on the occurrence of DM was 17%., Conclusions: An increasing prevalence of DM was observed, while diet/light/zero beverages consumption remains stable. A substantial reduction in the annual percentage change of DM could be observed if people stopped consuming diet/light soda/juice., (Copyright © 2023 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Anthropogenic activities have increased the dispersal of emerging contaminants (ECs), particularly of parabens, causing an escalation of their presence in wastewater (WW). Current WW technologies do not present satisfactory efficiency or sustainability in removing these contaminants. However, bioremediation with microalgae-based systems is proving to be a relevant technology for WW polishing, and the use of microalgae-bacteria consortia can improve the efficiency of WW treatment. This work aimed to study dual cultures of selected bacteria (Raoultella ornithinolytica, Acidovorax facilis, Acinetobacter calcoaceticus, Leucobacter sp. or Rhodococcus fascians) and the microalga Chlorella vulgaris in microbial growth and WW bioremediation - removal of methylparaben (MetP) and nutrients. The association with the bacteria was antagonistic for C. vulgaris biomass productivity as a result of the decreased growth kinetics in comparison to the axenic microalga. The presence of MetP did not disturb the growth of C. vulgaris under axenic or co-cultured conditions, except when associated with R. fascians, where growth enhancement was observed. The removal of MetP by the microalga was modest (circa 30 %, with a removal rate of 0.0343 mg/L.d), but increased remarkably when the consortia were used (> 50 %, with an average removal rate > 0.0779 mg/L.d), through biodegradation and photodegradation. For nutrient removal, the consortia were found to be less effective than the axenic microalga, except for nitrogen (N) removal by C. vulgaris w/ R. fascians. The overall results propose that C. vulgaris co-cultivation with bacteria can increase MetP removal, while negatively affecting the microalga growth and the consequent reduction of sludge production, highlighting the potential of microalgae-bacteria consortia for the effective polishing of WW contaminated with parabens., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

This work aimed to determine the ability of Listeria innocua (L.i.) to colonize eight materials found in food-processing and packaging settings and to evaluate the viability of the sessile cells. We also selected four commonly used phytochemicals (trans-cinnamaldehyde, eugenol, citronellol, and terpineol) to examine and compare their efficacies against L.i. on each surface. Biofilms were also deciphered in chamber slides using confocal laser scanning microscopy to learn more about how phytochemicals affect L.i. The materials tested were silicone rubber (Si), polyurethane (PU), polypropylene (PP), polytetrafluoroethylene (PTFE), stainless steel 316 L (SS), copper (Cu), polyethylene terephthalate (PET), and borosilicate glass (GL). L.i. colonized Si and SS abundantly, followed by PU, PP, Cu, PET, GL, and PTFE surfaces. The live/dead status ranged from 65/35% for Si to 20/80% for Cu, and the estimates of cells unable to grow on Cu were the highest, reaching even 43%. Cu was also characterized by the highest degree of hydrophobicity (ΔG TOT = -81.5 mJ/m 2 ). Eventually, it was less prone to attachment, as we could not recover L.i. after treatments with control or phytochemical solutions. The PTFE surface demonstrated the least total cell densities and fewer live cells (31%) as compared to Si (65%) or SS (nearly 60%). It also scored high in hydrophobicity degree (ΔG TOT = -68.9 mJ/m 2 ) and efficacy of phytochemical treatments (on average, biofilms were reduced by 2.1 log 10 CFU/cm 2 ). Thus, the hydrophobicity of surface materials plays a role in cell viability, biofilm formation, and then biofilm control and could be the prevailing parameter when designing preventive measures and interventions. As for phytochemical comparison, trans-cinnamaldehyde displayed greater efficacies, with the highest reductions seen on PET and Si (4.6 and 4.0 log 10 CFU/cm 2 ). The biofilms in chamber slides exposed to trans-cinnamaldehyde revealed the disrupted organization to a greater extent than other molecules. This may help establish better interventions via proper phytochemical selection for incorporation in environment-friendly disinfection approaches., Competing Interests: Declaration of Competing Interest 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 paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Ultraviolet irradiation C (UVC) has emerged as an effective strategy for microbial control in indoor public spaces. UVC is commonly applied for air, surface, and water disinfection. Unlike common 254 nm UVC, far-UVC at 222 nm is considered non-harmful to human health, being safe for occupied spaces, and still effective for disinfection purposes. Therefore, and allied to the urgency to mitigate the current pandemic of SARS-CoV-2, an increase in UVC-based technology devices appeared in the market with levels of pathogens reduction higher than 99.9 %. This environmentally friendly technology has the potential to overcome many of the limitations of traditional chemical-based disinfection approaches. The novel UVC-based devices were thought to be used in public indoor spaces such as hospitals, schools, and public transport to minimize the risk of pathogens contamination and propagation, saving costs by reducing manual cleaning and equipment maintenance provided by manpower. However, a lack of information about UVC-based parameters and protocols for disinfection, and controversies regarding health and environmental risks still exist. In this review, fundamentals on UVC disinfection are presented. Furthermore, a deep analysis of UVC-based technologies available in the market for the disinfection of public spaces is addressed, as well as their advantages and limitations. This comprehensive analysis provides valuable inputs and strategies for the development of effective, reliable, and safe UVC disinfection systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Coaggregation plays an important role in the development of multispecies biofilms in different environments, often serving as an active bridge between biofilm members and other organisms that, in their absence, would not integrate the sessile structure. The ability of bacteria to coaggregate has been reported for a limited number of species and strains. In this study, 38 bacterial strains isolated from drinking water (DW) were investigated for their ability to coaggregate, in a total of 115 pairs of combinations. Among these isolates, only Delftia acidovorans (strain 005P) showed coaggregating ability. Coaggregation inhibition studies have shown that the interactions mediating D. acidovorans 005P coaggregation were both polysaccharide-protein and protein-protein, depending on the interacting partner bacteria. Dual-species biofilms of D. acidovorans 005P and other DW bacteria were developed to understand the role of coaggregation on biofilm formation. Biofilm formation by Citrobacter freundii and Pseudomonas putida strains highly benefited from the presence of D. acidovorans 005P, apparently due to the production of extracellular molecules/public goods favouring microbial cooperation. This was the first time that the coaggregation capacity of D. acidovorans was demonstrated, highlighting its role in providing a metabolic opportunity for partner bacteria., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The first aim was to investigate the impact of different electromyography (EMG) parameters as a reference to normalize the EMG amplitude of the superficial quadriceps femoris muscles across different sets of a knee extension exercise. The second aim is to examine the reliability between days of the EMG parameters used as a reference. Eleven young males attended the laboratory on 4 different days and performed one repetition maximum test, maximumvoluntary isometric contractions, and a resistance training protocol until failure. Surface EMG was placed over the rectus femoris, vastus lateralis, and vastus medialis muscles. Seven EMG parameters were calculated from the tasks and used to normalize EMG amplitude measured during the resistance training protocol. A repeated-measures two-way ANOVA was used (normalized EMG amplitude × set) to compare normalized EMG across sets, while an intraclass correlation coefficient, coefficient of variation, and Bland-Altman plots were used to calculate the intra-day reliability of the EMG parameters. The present investigation showed that normalized EMG amplitude of the superficial muscles of the quadriceps measured during a knee extension exercise is influenced by the EMG parameter and depends on the muscle. While rectus femoris and vastus lateralis normalized EMG amplitude presented one parameter among seven showing similar value to the other parameters, VM showed two. Lastly, all EMG parameters for all muscles presented an overall excellent reliability and agreement between days., Competing Interests: Declaration of Competing Interest 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 paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Musk fragrances have been detected in drinking water (DW) at trace concentrations. However, their impact on the microbial quality of DW has been disregarded. This work provides a pioneer evaluation of the effects of two synthetic musks contaminants, tonalide (AHTN) and galaxolide (HHCB), in microbial biofilms formed on two different surfaces, polyvinyl chloride (PVC) and stainless steel AISI 316 (SS316). Three bacterial species isolated from DW (Acinetobacter calcoaceticus, Burkholderia cepacia and Stenotrophomonas maltophilia), were used to develop 7-day-old single and mixed species biofilms. The impact of musks was assessed directly on biofilms but also on the bacteria motility, biofilm formation ability and biofilm susceptibility to chlorination. AHTN musk caused the most remarkable effects by increasing the cellular density and viability of mixed biofilms, and the extracellular polysaccharides content of biofilms on SS316. Most of the alterations caused by the direct exposure of biofilms to musks were observed when SS316 was used as an adhesion surface. In contrast, the ability to form biofilms and their susceptibility to chlorine were more affected for bacteria from HHCB-exposed biofilms on PVC. The overall results demonstrate that the presence of musks at residual concentrations influences DW bacterial dynamics, with the potential to impact the DW quality and safety. The type of plumbing material may further impact the effects of musks., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Tuberculosis (TB) is a disease with worldwide presence and a major cause of death in several developing countries. Current diagnostic methodologies often lack specificity and sensitivity, whereas a long time is needed to obtain a conclusive result., Methods: In an effort to develop better diagnostic methods, this study aimed at the discovery of a biomarker signature for TB diagnosis using a Nuclear Magnetic Resonance based metabolomics approach. In this study, we acquired 1 H NMR spectra of blood serum samples of groups of healthy subjects, individuals with latent TB and of patients with pulmonary and extra-pulmonary TB. The resulting data were treated with uni- and multivariate statistical analysis., Results: Six metabolites (inosine, hypoxanthine, mannose, asparagine, aspartate and glutamate) were validated by an independent cohort, all of them related with metabolic processes described as associated with TB infection., Conclusion: The findings of the study are according with the WHO Target Product Profile recommendations for a triage test to rule-out active TB., Competing Interests: Conflicts of interest The authors declare no conflicts of interest., (Copyright © 2021 Chang Gung University. Published by Elsevier B.V. All rights reserved.)

Background: There is continued concern about residential proximity to agricultural pesticide use and possible adverse health effects. Studies on this subject have been scarce with inconsistent results. We explored associations between residential proximity to specific crops, pesticide use and cause-specific mortality in a prospective census-based cohort study in The Netherlands., Methods: Selecting inhabitants aged >30 living in less urbanized areas, at the same address for nine years up to baseline (2004) from a national register-based cohort, we followed ~3.1 million individuals for cause-specific mortality until 2012. We estimated the area of specific crop groups cultivated within buffers of 50 m, 100 m and 250 m around each individual's residence and the amount of fungicides, herbicides and insecticides used within the same buffers for the period 1995-2003. The association between these exposure proxies and 25 primary causes of death was investigated using Cox proportional hazards regression, adjusting for individual and area-level confounders., Results: Residential proximity to crops was associated with decreased mortality risks overall. In contrast to the overall trend an increased risk was observed for chronic lower respiratory diseases and proximity to maize cultivation. We found no evidence of an association between the amount of pesticides used and cause-specific mortality., Conclusions: In a large prospective census-based cohort study in The Netherlands we found evidence of an increased risk of chronic lower respiratory diseases in relation to maize cultivation which was not reflected in general pesticide use, hinting to specific pesticides or practices in maize cultivation that may lead to the observed increased risk., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The understanding of microbial susceptibility to disinfectants is an important step to provide drinking water (DW) of adequate microbiological quality. In drinking water distribution systems (DWDS) the application of disinfectants is the main approach to control microorganisms. Although chlorine has been commonly used for DW treatment, the increase of microbial resistance and the production of harmful disinfection by-products promote the necessity to seek new alternatives. This study evaluated the antimicrobial activity of sodium dichloroisocyanurate (NaDCC), trichloroisocyanuric acid (TCCA), and pentapotassium bis(peroxymonosulphate) bis(sulphate) (OXONE) against two emerging pathogens isolated from DW, Acinetobacter calcoaceticus and Stenotrophomonas maltophilia. Free chlorine from calcium hypochlorite was used for comparison. The dose and time-responses against planktonic bacteria were performed as well as the assessment of the effects on membrane integrity. Moreover, the effects against 48 h-old biofilms formed on polyvinyl chloride and stainless steel were evaluated in terms of biofilm culturability and removal. Minimum bactericidal concentrations of 2.1 and 3.1 mg/L for NaDCC, 2.5 and 3.8 mg/L for TCCA, 340 and 690 mg/L for OXONE, and 0.80 and 1.0 mg/L for free chlorine alone were obtained against S. maltophilia and A. calcoaceticus, respectively. The kinetic modeling revealed that NaDCC and TCCA caused similar inactivation rates and the time for first log reduction by OXONE was less than 10 min, for both bacteria. All the disinfectants triggered significant bacterial cytoplasmic membrane destabilization, even at sub-lethal concentrations. A 30 min treatment with the disinfectants allowed a reduction in the biofilm culturability up to 5 log. OXONE was the disinfectant with the best efficiency against both bacterial biofilms. However, none of the disinfectants caused significant biofilm removal (reduction < 1 log cells/cm 2 ). This study highlights NaDCC, TCCA, and OXONE as promising alternatives to free chlorine for DW disinfection, particularly for planktonic growth control and biofilm culturability reduction., Competing Interests: Declaration of competing interest 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 paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The presence of contaminants of emerging concern (CECs) in the environment has been recognized as a worldwide concern. In particular, water pollution by CECs is becoming a major global problem, which requires ongoing evaluation of water resources policies at all levels and the use of effective and innovative wastewaters treatment processes for their removal. Microalgae have been increasingly recognized as relevant for wastewater polishing, including CECs removal. These microorganisms are commonly cultivated in suspension. However, the use of planktonic microalgae for wastewater treatment has limitations in terms of microbiological contamination, process effectiveness and sustainability. The use of consortia of microalgae and bacteria represents a significant advance for sustainable wastewater polishing, particularly when the microorganisms are associated as biofilms. These immobilized mixed cultures can overcome the limitations of suspended-microalgae systems and improve the performance of the involved species for CECs removal. In addition, microalgae-bacteria based systems can offer a relevant combined effect for CECs removal and biomass production enhancement. This study reviews the advantages and advances on the use of microalgae for wastewater treatment, highlighting the potential on the use of microalgae-bacteria biofilms for CECs removal and the further biomass valorisation for third-generation biofuel production., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Vascular endothelial growth factor A (VEGF-A) is expressed by several cell types and is a crucial factor for angiogenic-osteogenic coupling. However, the immunolocalization of VEGF-A during the early stages of the alveolar process formation remains underexplored. Thus, we analyzed the spatio-temporal immunolocalization of VEGF-A and its relationship with Runt-related transcription factor 2 (Runx2) and osterix (Osx) during the early steps of intramembranous ossification of the alveolar process in rat embryos. Embryo heads (E) of 16, 18 and 20-day-old rats were processed for paraffin embedding. Histomorphometry and immunohistochemistry to detect VEGF-A, Runx2, and Osx (osteoblast differentiation markers) were performed. The volume density of bone tissue including bone cells and blood vessels increased significantly in E18 and E20. Cells showing high VEGF-A immunoreactivity were initially observed within a perivascular niche in the ectomesenchyme; afterwards, these cells were diffusely located near bone formation sites. Runx2-and Osx-immunopositive cells were observed in corresponded regions of cells showing strong VEGF-A immunoreactivity. Although these immunostained cells were observed in all specimens, this immunolocalization pattern was more evident in E16 specimens and gradually decreased in E18 and E20 specimens. Double immunofluorescence labelling showed intracellular co-localization of Osx and VEGF-A in cells surrounding the developing alveolar process, indicating a crucial role of VEGF-A in osteoblast differentiation. Our results showed VEGF-A immunoexpression in osteoblasts and its precursors during the maxillary alveolar process formation of rat embryos. Moreover, the VEGF-A-positive cells located within a perivascular niche at the early stages of the alveolar process development suggest a crosstalk between endothelium and ectomesenchymal cells, reinforcing the angiogenic-osteogenic coupling in this process., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2021. Published by Elsevier Inc.)

The treatment of Parkinson´s disease (PD) has benefited from significant advances resulting from the increasing research efforts focused on new therapeutics. However, the current treatments for PD are mostly symptomatic, alleviating disease symptoms without reversing or retarding disease progression. Thus, it is critical to find new molecules that can result in more effective treatments. Within this framework, this study aims to evaluate the neuroprotective and anti-inflammatory effects of three compounds (eleganolone, eleganonal and fucosterol) isolated from the brown seaweed Bifurcaria bifurcata. In vitro neuroprotective effects were evaluated on a PD cellular model induced by the neurotoxin 6-hydroxydopamine (6-OHDA) on SH-SY5Y human cells, while lipopolysaccharide (LPS) - stimulated RAW 264.7 macrophages were used to evaluate the anti-inflammatory potential. Additionally, the underlying mechanisms of action were also investigated. Compounds were isolated by preparative chromatographic methods and their structural elucidation attained by NMR spectroscopy. Among the tested compounds, eleganolone (0.1-1 µM; 24 h) reverted the neurotoxicity induced by 6-OHDA in about 20%. The neuroprotective effects were mediated by mitochondrial protection, reduction of oxidative stress, inflammation and apoptosis, and inhibition of NF-kB pathway. The results suggest that eleganolone may provide advantages in the treatment of neurodegenerative conditions and, therefore, should be considered for future preclinical studies., (Copyright © 2021. Published by Elsevier Ltd.)

Recent studies suggest that temporoparietal junction (TPJ) modulation can influence attention and social cognition performance. Nevertheless, no studies have used multichannel transcranial direct current stimulation (tDCS) over bilateral TPJ to estimate the effects on these neuropsychological functions. The project STIPED is using optimized multichannel stimulation as an innovative treatment approach for chronic pediatric neurodevelopmental disorders, namely in children/adolescents with Autism Spectrum Disorder (ASD). In this pilot study, we aim to explore whether anodal multichannel tDCS coupled with a Joint Attention Task (JAT) influences social-cognitive task performance relative to sham stimulation, both in an Emotion Recognition Task (ERT) and in a Mooney Faces Detection Task (MFDT), as well as to evaluate this technique's safety and tolerability. Twenty healthy adults were enrolled in a randomized, single-blinded, sham-controlled, crossover study. During two sessions, participants completed the ERT and the MFDT before and after 20min of sham or anodal tDCS over bilateral TPJ. No significant differences on performance accuracy and reaction time were found between stimulation conditions for all tasks, including the JAT. A significant main time effect for overall accuracy and reaction time was found for the MFDT. Itching was the most common side effect and stimulation conditions detection was at chance level. Results suggest that multichannel tDCS over bilateral TPJ does not affect performance of low-level emotional recognition tasks in healthy adults. Although preliminary safety and tolerability are demonstrated, further studies over longer periods will be pursued to investigate the clinical efficacy in children/adolescents with ASD, where social cognition impairments are preponderant., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Pediatric hypertension is typically defined as blood pressure ≥ sex-, age-, and height-specific 95th percentile (high) cutoffs. Given the number of strata, there are hundreds of cutoffs for defining elevated and high blood pressure that make it cumbersome to use in clinical practice. This study aimed to evaluate the utility of the static cutoffs for pediatric hypertension (120/80 mm Hg for children and 130/80 mm Hg for adolescents) in determining high carotid intimamedia thickness (cIMT) in children and adolescents., Methods: Data were from 6 population-based cross-sectional studies in Brazil, China, Greece, Italy, Spain, and the United Kingdom. A total of 4280 children and adolescents, aged 6 to 17 years, were included. High cIMT was defined as cIMT ≥ sex-, age- and cohort-specific 90th percentile cutoffs., Results: Compared with normal blood pressure, hypertension defined using the percentile-based cutoffs from 2017 American Academy of Pediatrics guideline, and the static cutoffs were associated with similar higher odds for high cIMT (percentile-based cutoffs: odds ratio [OR], 1.46, 95% confidence interval [CI], 1.15-1.86; static cutoffs: OR, 1.65, 95% CI, 1.25-2.17), after adjustment for sex, age, race/ethnicity, body mass index, high-density lipoprotein-cholesterol, triglyceride, and fasting blood glucose. The similar utility of 2 definitions in determining high cIMT was further confirmed by area under the receiver operating characteristic curve and net reclassification improvement methods (P for difference > 0.05)., Conclusion: Static cutoffs (120/80 mm Hg for children, 130/80 mm Hg for adolescents) performed similarly compared with percentile-based cutoffs in determining high cIMT, supporting the use of static cutoffs in identifying pediatric hypertension in clinical practice., (Copyright © 2020 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.)

Microbial fuel cells (MFCs) have been receiving an increased attention over the last years due to their potential to combat two global problems: waste pollution and energy demand. Additionally, when a wastewater is used, MFCs can perform its treatment while recovering energy, leading to the possibility of energy-producing wastewater treatment plants, offsetting their operational costs. However, to overcome their current limitations (lower power outputs and higher costs), a clear understanding of the effect of operation and design parameters on its overall performance is mandatory. Therefore, the goal of this work was to evaluate the effect of operating conditions - batch cycle and yeast extract concentration, and design parameters - anode electrode area, membrane thickness and active area, on the overall performance of a single chamber MFC. The MFC operated with a pure culture of Lactobacillus pentosus and a synthetic wastewater based on a real dairy industry effluent. The overall performance was evaluated through the power output and the COD removal rate. Additionally, the biofilm formed at the anode electrode was characterized in terms of biomass, proteins and polysaccharides content. For the conditions used in this work, a maximum power density of 5.04 ± 0.39 mW/m 2 was achieved with an anode electrode area of 61 cm 2 , a batch cycle of 48 h, 50 mg/L of yeast extract and a Nafion 212 membrane with an active area of 25 cm 2 . The different conditions tested had a clear effect on the MFC energy production and biofilm characteristics, but not on the ability of L. pentosus to treat the dairy wastewater. The COD removal rates were in the range between 42% and 58%, for all the conditions tested., (Copyright © 2018. Published by Elsevier B.V.)

Concerns on the presence of emerging contaminants (ECs) in water sources have increased in recent years. The lack of efficient technologies to remove ECs from residual waters contributes for their appearance in drinking water distribution systems (DWDS). Therefore, sessile microorganisms on DWDS pipes are continuously exposed to trace concentrations of ECs. However, no data exists on the role of ECs on the resident microbiota. The present work aims to understand the effects of prolonged exposure of a bacterial strain of Stenotrophomonas maltophilia, isolated from a DWDS, in both planktonic and biofilm states, to trace concentrations of selected ECs (antipyrine-ANTP; diclofenac sodium salt-DCF; ibuprofen-IBP; galaxolide-GAL; tonalide-TON; carbamazepine-CBZ; clofibric acid-CA; tylosin-TY) on its tolerance to sodium hypochlorite (NaOCl) and resistance to antibiotics. Pre-established S. maltophilia biofilms were exposed to ECs for 26 d. Subsequently, the planktonic behaviour of the biofilm cells grown in the presence of ECS was characterized in terms of susceptibility to NaOCl and to selected antibiotics (levofloxacin and trimethoprim-sulfamethoxazole). Moreover, S.maltophilia was tested on its biofilm productivity in the presence of ECs (alone and mixed). These biofilms were challenged by NaOCl in order to assess the role of ECs on biofilm susceptibility. The results did not evidence remarkable effects of ECs on planktonic S. maltophilia susceptibility to NaOCl and antibiotics. However, S. maltophilia biofilm production and susceptibility to NaOCl was affected from ECs pre-exposure, particularly by the combination of different ECs (CA + CBZ, CA + IBP, CA + CBZ + IBP). S. maltophilia biofilms became more resistant to removal by NaOCl when developed in the presence of mixtures of CA + CBZ and CA + CBZ + IBP. Also, biofilm production was significantly affected. CA was present in all the combinations that altered biofilm behaviour. The overall results propose that exposure to ECs for 26 days had not a huge impact on S. maltophilia planktonic antimicrobial susceptibility. Nevertheless, the prolonged exposure to some ECs altered biofilm production and tolerance to NaOCl, with a potential practical outcome of hindering DWDS disinfection. The simultaneous presence of different ECs in the environment may amplify biofilm resilience., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Iron deficiency is the most common nutritional deficit worldwide. The goal of this work was to obtain iron-pectin beads by ionic gelation and evaluate their physiological behavior to support their potential application in the food industry. The beads were firstly analyzed by scanning electronic microscopy, and then physical-chemically characterized by performing swelling, thermogravimetric, porosimetry, Mössbauer spectroscopy and X-ray fluorescence analyses, as well as by determining the particle size. Then, physiological assays were carried out by exposing the beads to simulated gastric and intestinal environments, and determining the iron absorption and transepithelial transport into Caco-2/TC7 cells. Iron-pectin beads were spherical (diameter 1-2 mm), with high density (1.29 g/mL) and porosity (93.28%) at low pressure, indicating their high permeability even when exposed to low pressure. Swelling in simulated intestinal medium (pH 8) was higher than in simulated gastric medium. The source of iron [FeSO 4 (control) or iron-pectin beads] did not have any significant effect on the mineral absorption. Regarding transport, the iron added to the apical pole of Caco-2/TC7 monolayers was recovered in the basal compartment, and this was proportional with the exposure time. After 4 h of incubation, the transport of iron arising from the beads was significantly higher than that of the iron from the control (FeSO 4 ). For this reason, iron-pectin beads appear as an interesting system to overcome the low efficiency of iron transport, being a potential strategy to enrich food products with iron, without altering the sensory properties., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The presence of biofilms in drinking water distribution systems (DWDS) is a global public health concern as they can harbor pathogenic microorganisms. Sodium hypochlorite (NaOCl) is the most commonly used disinfectant for microbial growth control in DWDS. However, its effect on biofilm removal is still unclear. This work aims to evaluate the effects of the combination of chemical (NaOCl) and mechanical stresses on the removal of single and dual species biofilms of two bacteria isolated from DWDS and considered opportunistic, Acinectobacter calcoaceticus and Stenotrophomonas maltophilia. A rotating cylinder reactor was successfully used for the first time in drinking water biofilm studies with polyvinyl chloride as substratum. The single and dual species biofilms presented different characteristics in terms of metabolic activity, mass, density, thickness and content of proteins and polysaccharides. Their complete removal was not achieved even when a high NaOCl concentrations and an increasing series of shear stresses (from 2 to 23Pa) were applied. In general, NaOCl pre-treatment did not improve the impact of mechanical stress on biofilm removal. Dual species biofilms were colonized mostly by S. maltophilia and were more susceptible to chemical and mechanical stresses than these single species. The most efficient treatment (93% biofilm removal) was the combination of NaOCl at 175mg·l -1 with mechanical stress against dual species biofilms. Of concern was the high tolerance of S. maltophilia to chemical and mechanical stresses in both single and dual species biofilms. The overall results demonstrate the inefficacy of NaOCl on biofilm removal even when combined with high shear stresses., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Liposomes are exceptional carriers for therapeutic drug delivery. However, they generally suffer from poor cell penetration, low half-life in bloodstream and loss of functionality during storage. To overcome these problems some strategies can be applied, such as functionalization with polymers and the use of protective molecules during dehydration processes. This work reports a complete study about the stability, including freeze-drying in the presence of trehalose, storage and internalization into HEp-2 cells, of stable formulations of pH sensitive polymer-liposome complexes (PLC) composed of soybean lecithin and crosslinked/non-crosslinked poly(acrylic acid) with a cholesterol end-group (CHO-PAA). The results showed that the average hydrodynamic particle size of the complexes persisted unaffected for approximately 75 days after freeze-drying in the presence of 10% w/v trehalose. The efficiency of calcein encapsulation and release profiles in physiologic conditions exhibited no significant alterations when stored for 0 and 1 month, and for 2 and 3 months of storage the calcein release increased with time. The stored complexes were efficiently uptaken into HEp-2-cells, as determined by confocal microscopy. In all cases, the percentage of viable cells was above 90%, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, indicating no potential toxicity. Finally, transepithelial transport assays demonstrated that both fresh and 2 months-stored complexes could transport their calcein content through HEp-2 monolayers over time., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Technological innovations in vaccinology have recently contributed to bring about novel insights for the vaccine-induced immune response. While the current protocols that use peripheral blood samples may provide abundant data, a range of distinct components of whole blood samples are required and the different anticoagulant systems employed may impair some properties of the biological sample and interfere with functional assays. Although the interference of heparin in functional assays for viral neutralizing antibodies such as the functional plaque-reduction neutralization test (PRNT), considered the gold-standard method to assess and monitor the protective immunity induced by the Yellow fever virus (YFV) vaccine, has been well characterized, the development of pre-analytical treatments is still required for the establishment of optimized protocols. The present study intended to optimize and evaluate the performance of pre-analytical treatment of heparin-collected blood samples with ecteola-cellulose (ECT) to provide accurate measurement of anti-YFV neutralizing antibodies, by PRNT. The study was designed in three steps, including: I. Problem statement; II. Pre-analytical steps; III. Analytical steps. Data confirmed the interference of heparin on PRNT reactivity in a dose-responsive fashion. Distinct sets of conditions for ECT pre-treatment were tested to optimize the heparin removal. The optimized protocol was pre-validated to determine the effectiveness of heparin plasma:ECT treatment to restore the PRNT titers as compared to serum samples. The validation and comparative performance was carried out by using a large range of serum vs heparin plasma:ECT 1:2 paired samples obtained from unvaccinated and 17DD-YFV primary vaccinated subjects. Altogether, the findings support the use of heparin plasma:ECT samples for accurate measurement of anti-YFV neutralizing antibodies., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Fourier transform infrared (FT-IR) microscopy was used to image tissue samples from twenty patients diagnosed with thyroid carcinoma. The spectral data were then used to differentiate between follicular thyroid carcinoma and follicular variant of papillary thyroid carcinoma using principle component analysis coupled with linear discriminant analysis and a Naïve Bayesian classifier operating on a set of computed spectral metrics. Classification of patients' disease type was accomplished by using average spectra from a wide region containing follicular cells, colloid, and fibrosis; however, classification of disease state at the pixel level was only possible when the extracted spectra were limited to follicular epithelial cells in the samples, excluding the relatively uninformative areas of fibrosis. The results demonstrate the potential of FT-IR microscopy as a tool to assist in the difficult diagnosis of these subtypes of thyroid cancer, and also highlights the importance of selectively and separately analyzing spectral information from different features of a tissue of interest.

The development of polymer-liposome complexes (PLCs), in particular for biomedical applications, has grown significantly in the last decades. The importance of these studies comes from the emerging need in finding intelligent controlled release systems, more predictable, effective and selective, for applications in several areas, such as treatment and/or diagnosis of cancer, neurological, dermatological, ophthalmic and orthopedic diseases, gene therapy, cosmetic treatments, and food engineering. This work reports the development and characterization of a pH sensitive system for controlled release based on PLCs. The selected hydrophilic polymer was poly(acrylic acid) (PAA) synthesized by atom transfer radical polymerization (ATRP) with a cholesterol (CHO) end-group to improve the anchoring of the polymer into the lipid bilayer. The polymer was incorporated into liposomes formulated from soybean lecithin and stearylamine, with different stearylamine/phospholipid and polymer/phospholipid ratios (5, 10 and 20%). The developed PLCs were characterized in terms of particle size, polydispersity, zeta potential, release profiles, and encapsulation efficiency. Cell viability studies were performed to assess the cytotoxic potential of PLCs. The results showed that the liposomal formulation with 5% of stearylamine and 10% of polymer positively contribute to the stabilization of the complexes. Afterwards, the carboxylic acid groups of the polymer present at the surface of the liposomes were crosslinked and the same parameters analyzed. The crosslinked complexes showed to be more stable at physiologic conditions. In addition, the release profiles at different pHs (2-12) revealed that the obtained complexes released all their content at acidic conditions. In summary, the main accomplishments of this work are: (i) innovative synthesis of cholesterol-poly(acrylic acid) (CHO-PAA) by ATRP; (ii) stabilization of the liposomal formulation by incorporation of stearylamine and CHO-PAA; (iii) new approach for CHO-PAA crosslinking, resulting in more stable PLCs at physiological conditions; (iv) destabilization of PLCs upon slight changes of pH, showing their pH sensitivity; and (v) the PLCs do not exhibit cellular toxicity., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Mitochondrial DNA (mtDNA) mutations have been assigned as a major cause of genetic disease. When a novel sequence variation is found, it is necessary to evaluate its functional impact, usually requiring functional molecular studies. Given the fact that this approach is difficult to put in practice in a routine basis, it is possible to take advantage of the in silico tools available and predict protein/RNA structure changes and therefore pathogenicity. Here, we describe the characterization of five undescribed mtDNA variants, upon detection of 23 unclassified alterations at Laboratory of Biochemical Genetics, from 2004 to 2014. Those five sequence variations are located in protein-coding genes, in five patients with a diverse range of mitochondrial respiratory chain disease phenotypes including encephalopathy, optic neuropathy, developmental delay, deafness and epilepsy. According to the prediction established by in silico analysis using tools to predict structure and function changes (ClustalW2 ® , PolyPhen-2 ® , SIFT ® , MutationAssessor ® , PredictProtein ® , Provean ® , I-TASSER ® , Haplogrep ® ), from the 23 variants analyzed, the five described are potentially pathogenic. This approach is inexpensive and compatible with a rapid first line response to clinical demanding, contributing to a more rationale genetic diagnosis concerning novel mutations and to clarify the mtDNA involvement in these pathologies., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

The aim of this paper was to test the factorial structure and evaluate the psychometric properties of the Youth Psychopathic Traits Inventory (YPI). The YPI is composed of 10 dimensions that further represent three hypothesized facets of the classical description of psychopathy: callousness, interpersonal manipulation and impulsiveness. A sample of 500 adolescents aged 12 to 18 (M = 14.87; SD = 1.67) from northern Portugal participated in this study. The results generally confirmed the factorial structure of the YPI in this sample, with some qualifications., (Copyright © 2015 The Foundation for Professionals in Services for Adolescents. Published by Elsevier Ltd. All rights reserved.)

Recalcitrant polycyclic aromatic hydrocarbons (PAHs) released into seawater end up in the deep sea sediments (DSSs). However, their fate here is often oversimplified by theoretical models. Biodegradation of PAHs in DSSs, is assumed to be similar to biodegradation in surface habitats, despite high hydrostatic pressures and low temperatures that should significantly limit PAH biodegradation. Bacteria residing in the DSSs (related mainly to α- and γ-Proteobacteria) have been shown to or predicted to possess distinct genes, enzymes and metabolic pathways, indicating an adaptation of these bacterial communities to the psychro-peizophilic conditions of the DSSs. This work summarizes some of the most recent research on DSS hydrocarbonoclastic populations and mechanisms of PAH degradation and discusses the challenges posed by future high CO2 and UV climate scenarios on biodegradation of PAHs in DSSs., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A tumor is a heterogeneous population of cells that provides an environment in which every cell resides in a microenvironmental niche. Microscopic evaluation of tissue sections, based on histology and immunohistochemistry, has been a cornerstone in pathology for decades. However, the dawn of novel technologies to investigate genetic aberrations is currently adopted in routine molecular pathology. We herein describe our view on how recent developments in molecular technologies, focusing on proximity ligation assay and padlock probes, can be applied to merge the two branches of pathology, allowing molecular profiling under histologic observation. We also discuss how the use of image analysis will be pivotal to obtain information at a cellular level and to interpret holistic images of tissue sections. By understanding the cellular communications in the microecology of tumors, we will be at a better position to predict disease progression and response to therapy., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The adhesion of Escherichia coli to glass and polydimethylsiloxane (PDMS) at different flow rates (between 1 and 10 ml s(-1)) was monitored in a parallel plate flow chamber in order to understand the effect of surface properties and hydrodynamic conditions on adhesion. Computational fluid dynamics was used to assess the applicability of this flow chamber in the simulation of the hydrodynamics of relevant biomedical systems. Wall shear stresses between 0.005 and 0.07 Pa were obtained and these are similar to those found in the circulatory, reproductive and urinary systems. Results demonstrate that E. coli adhesion to hydrophobic PDMS and hydrophilic glass surfaces is modulated by shear stress with surface properties having a stronger effect at the lower and highest flow rates tested and with negligible effects at intermediate flow rates. These findings suggest that when expensive materials or coatings are selected to produce biomedical devices, this choice should take into account the physiological hydrodynamic conditions that will occur during the utilization of those devices., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The reactivation or reexpression of telomerase (TERT) is a widespread feature of neoplasms. TERT promoter mutations were recently reported that were hypothesized to result from UV radiation. In this retrospective study, we assessed TERT promoter mutations in 196 cutaneous basal cell carcinomas (BCCs), including 102 tumors from X-irradiated patients, 94 tumors from patients never exposed to ionizing radiation treatment, and 116 melanomas. We sought to evaluate the effects of UV and X-ray irradiation on TERT mutation frequency. TERT mutations were detected in 27% of BCCs from X-irradiated patients, 51% of BCCs from nonirradiated patients, and 22% of melanoma patients. TERT mutations were significantly increased in non-X-irradiated BCC patients compared with X-irradiated BCC patients; the mutations also presented a different mutation signature. In nonirradiated patients, TERT mutations were more frequent in BCCs of sun-exposed skin, supporting a possible causative role of UV radiation. In melanoma, TERT promoter mutations were generally restricted to intermittent sun-exposed areas and were associated with nodular and superficial spreading subtypes, increased thickness, ulceration, increased mitotic rate, and BRAFV600E mutations. Our results suggest that various carcinogenic factors may cause distinct TERT promoter mutations in BCC and that TERT promoter mutations might be associated with a poorer prognosis in melanoma.

The prevalence of piroplasm (order Piroplasmida) infection was assessed in blood and bone marrow samples from 91 red foxes (Vulpes vulpes) from northern, central and southern Portugal by means of molecular methods. PCR for the 18S rRNA gene of Babesia spp. followed by sequencing revealed 63 foxes positive for the Babesia microti-like piroplasm (syn. Theileria annae) (69.2%; 95% confidence interval [CI]: 58.7-78.5%) and one fox positive for Babesia canis (1.1%; 95% CI: 0.0-6.0%). Positivity to the B. microti-like piroplasm or B. canis in 43 blood samples (83.7%) was significantly higher (p<0.001) than in 43 paired bone marrow samples (20.9%). There were no statistically significant differences in the prevalence of infection between genders (p=0.219) or age groups (<2 years vs. ≥ 2 years) (p=1.0). This is the first report of the B. microti-like piroplasm in foxes from Portugal as well as the first report on detection by PCR and genotyping of B. canis in a red fox worldwide. A natural cycle of the B. microti-like piroplasm is suggested in red fox populations based on the high prevalence of the protozoan. Red foxes might be a reservoir of the B. microti-like piroplasm and a source of infection to dogs., (Copyright © 2013 Elsevier B.V. All rights reserved.)