Search results for: Augusto Barata Da Rocha

Authors: Danny Alexander R. Moreno, Gustavo L. Sartorello, Yuli Andrea P. Bermudez, Richard R. Lobo, Ives Claudio S. Bueno, Augusto H. Gameiro

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Meat production is a significant sector for Brazil's economy; however, the agricultural segment has suffered censure regarding the negative impacts on the environment, which consequently results in climate change. Therefore, it is essential the implementation of nutritional strategies that can improve the environmental performance of livestock. This research aimed to estimate the environmental impact and profitability of the use of yerba mate extract (Ilex paraguariensis) as an additive in the feeding of feedlot lamb. Thirty-six castrated male lambs (average weight of 23.90 ± 3.67 kg and average age of 75 days) were randomly assigned to four experimental diets with different levels of inclusion of yerba mate extract (0, 1, 2, and 4 %) based on dry matter. The animals were confined for fifty-three days and fed with 60:40 corn silage to concentrate ratio. As an indicator of environmental impact, the carbon footprint (CF) was measured as kg of CO₂ equivalent (CO₂-eq) per kg of body weight produced (BWP). The greenhouse gas (GHG) emissions such as methane (CH₄) generated from enteric fermentation, were calculated using the sulfur hexafluoride gas tracer (SF₆) technique; while the CH₄, nitrous oxide (N₂O - emissions generated by feces and urine), and carbon dioxide (CO₂ - emissions generated by concentrate and silage processing) were estimated using the Intergovernmental Panel on Climate Change (IPCC) methodology. To estimate profitability, the gross margin was used, which is the total revenue minus the total cost; the latter is composed of the purchase of animals and food. The boundaries of this study considered only the lamb fattening system. The enteric CH₄ emission from the lamb was the largest source of on-farm GHG emissions (47%-50%), followed by CH₄ and N₂O emissions from manure (10%-20%) and CO₂ emission from the concentrate, silage, and fossil energy (17%-5%). The treatment that generated the least environmental impact was the group with 4% of yerba mate extract (YME), which showed a 3% reduction in total GHG emissions in relation to the control (1462.5 and 1505.5 kg CO₂-eq, respectively). However, the scenario with 1% YME showed an increase in emissions of 7% compared to the control group. In relation to CF, the treatment with 4% YME had the lowest value (4.1 kg CO₂-eq/kg LW) compared with the other groups. Nevertheless, although the 4% YME inclusion scenario showed the lowest CF, the gross margin decreased by 36% compared to the control group (0% YME), due to the cost of YME as a food additive. The results showed that the extract has the potential for use in reducing GHG. However, the cost of implementing this input as a mitigation strategy increased the production cost. Therefore, it is important to develop political strategies that help reduce the acquisition costs of input that contribute to the search for the environmental and economic benefit of the livestock sector.

Keywords: meat production, natural additives, profitability, sheep

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Authors: Raphael C. Zero, Marita V. Cardozo, Thiago A. S. S. Rocha, Mariana T. Kihara, Fernando A. Ávila, Fabrício S. Oliveira

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There are several fixative and preservative solutions for use on cadavers, many of them using formaldehyde as the fixative or anatomical part preservative. In some countries, such as Brazil, this toxic agent has been increasingly restricted. The objective of this study was to microbiologically identify and quantify the key agents in tanks containing 96GL ethanol or sodium chloride solutions, used respectively as fixatives and preservatives of cat cadavers. Eight adult cat corpses, three females and five males, with an average weight of 4.3 kg, were used. After injection via the external common carotid artery (120 ml/kg, 95% 96GL ethyl alcohol and 5% pure glycerin), the cadavers were fixed in a plastic tank with 96GL ethanol for 60 days. After fixing, they were stored in a 30% sodium chloride aqueous solution for 120 days in a similar tank. Samples were collected at the start of the experiment - before the animals were placed in the ethanol tanks, and monthly thereafter. The bacterial count was performed by Pour Plate Method in BHI agar (Brain Heart Infusion) and the plates were incubated aerobically and anaerobically for 24h at 37ºC. MacConkey agar, SPS agar (Sulfite Polymyxin Sulfadizine) and MYP Agar Base were used to isolate the microorganisms. There was no microbial growth in the samples prior to alcohol fixation. After 30 days of fixation in the alcohol solution, total aerobic and anaerobic (<1.0 x 10 CFU/ml) were found and Pseudomonas sp., Staphylococcus sp., Clostridium sp. were the identified agents. After 60 days in the alcohol fixation solution, total aerobes (<1.0 x 10 CFU/ml) and total anaerobes (<2.2 x 10 CFU/mL) were found, and the identified agents were the same. After 30 days of storage in the aqueous solution of 30% sodium chloride, total aerobic (<5.2 x 10 CFU/ml) and total anaerobes (<3.7 x 10 CFU/mL) were found and the agents identified were Staphylococcus sp., Clostridium sp., and fungi. After 60 days of sodium chloride storage, total aerobic (<3.0 x 10 CFU / ml) and total anaerobes (<7.0 x 10 CFU/mL) were found and the identified agents remained the same: Staphylococcus sp., Clostridium sp., and fungi. The microbiological count was low and visual inspection did not reveal signs of contamination in the tanks. There was no strong odor or purification, which proved the technique to be microbiologically effective in fixing and preserving the cat cadavers for the four-month period in which they are provided to undergraduate students of University of Veterinary Medicine for surgery practice. All experimental procedures were approved by the Municipal Legal Department (protocol 02.2014.000027-1). The project was funded by FAPESP (protocol 2015-08259-9).

Keywords: anatomy, fixation, microbiology, small animal, surgery

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Authors: Jesús Horacio, Alfredo Ollero, Víctor Bouzas-Blanco, Augusto Pérez-Alberti

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Rivers have traditionally been classified, assessed and managed in terms of hydrological, chemical and / or biological criteria. Geomorphological classifications had in the past a secondary role, although proposals like River Styles Framework, Catchment Baseline Survey or Stroud Rural Sustainable Drainage Project did incorporate geomorphology for management decision-making. In recent years many studies have been attracted to the geomorphological component. The geomorphological processes and their associated forms determine the structure of a river system. Understanding these processes and forms is a critical component of the sustainable rehabilitation of aquatic ecosystems. The fluvial auto-classification approach suggests that a river is a self-built natural system, with processes and forms designed to effectively preserve their ecological function (hydrologic, sedimentological and biological regime). Fluvial systems are formed by a wide range of elements with multiple non-linear interactions on different spatial and temporal scales. Besides, the fluvial auto-classification concept is built using data from the river itself, so that each classification developed is peculiar to the river studied. The variables used in the classification are specific stream power and mean grain size. A discriminant analysis showed that these variables are the best characterized processes and forms. The statistical technique applied allows to get an individual discriminant equation for each geomorphological type. The geomorphological classification was developed using sites with high naturalness. Each site is a control point of high ecological and geomorphological quality. The changes in the conditions of the control points will be quickly recognizable, and easy to apply a right management measures to recover the geomorphological type. The study focused on Galicia (NW Spain) and the mapping was made analyzing 122 control points (sites) distributed over eight river basins. In sum, this study provides a method for fluvial geomorphological classification that works as an open and flexible tool underlying the fluvial auto-classification concept. The hydrographic mapping is the visual expression of the results, such that each river has a particular map according to its geomorphological characteristics. Each geomorphological type is represented by a particular type of hydraulic geometry (channel width, width-depth ratio, hydraulic radius, etc.). An alteration of this geometry is indicative of a geomorphological disturbance (whether natural or anthropogenic). Hydrographic mapping is also dynamic because its meaning changes if there is a modification in the specific stream power and/or the mean grain size, that is, in the value of their equations. The researcher has to check annually some of the control points. This procedure allows to monitor the geomorphology quality of the rivers and to see if there are any alterations. The maps are useful to researchers and managers, especially for conservation work and river restoration.

Keywords: fluvial auto-classification concept, mapping, geomorphology, river

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Authors: Ian Christopher Rocha

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This study aimed to determine the association of the quality of life (QOL) of the caregivers of children in need of special protection (CNSP) in child-caring institutions in Metro Manila with the levels of their role overload (RO) and role distress (RD). The CNSP in this study covered the orphaned, abandoned, abused, neglected, exploited, and mentally-challenged children. In this study, the domains of QOL included physical health (PH), psychological health, social health (SH), and living conditions (LC). It also intended to ascertain the association of their personal and work-related characteristics with their RO and RD levels. The respondents of this study were 130 CNSP caregivers in 17 residential child-rearing institutions in Metro Manila. A purposive non-probability sampling was used. Using a quantitative methodological approach, the survey method was utilized to gather data with the use of a self-administered structured questionnaire. Data were analyzed using both descriptive and inferential statistics. Results revealed that the level of RO, the level of RD, and the QOL of the CNSP caregivers were all moderate. Data also suggested that there were significant positive relationships between the RO level and the caregivers’ characteristics, such as age, the number of training, and years of service in the institution. At the same time, the findings revealed that there were significant positive relationships between the RD level and the caregivers’ characteristics, such as age and hours of care rendered to their care recipients. In addition, the findings suggested that all domains of their QOL obtained significant relationships with their RO level. For the correlations of their level of RO and their QOL domains, the PH and the LC obtained a moderate negative correlation with the RO level while the rest of the domains obtained weak negative correlations with RO level. For the correlations of their level of RD and the QOL domains, all domains, except SH, obtained strong negative correlations with the level of RD. The SH revealed to have a moderate negative correlation with RD level. In conclusion, caregivers who are older experience higher levels of RO and RD; caregivers who have more training and years of service experience the higher level of RO; and caregivers who have longer hours of rendered care experience the higher level of RD. In addition, the study affirmed that if the levels of RO and RD are high, the QOL is low, and vice versa. Therefore, the RO and RD levels are reliable predictors of the caregivers’ QOL. In relation, the caregiving situation in the Philippines revealed to be unique and distinct from other countries because the levels of RO and RD and the QOL of Filipino CNSP caregivers were all moderate in contrast with their foreign counterparts who experience high caregiving RO and RD leading to low QOL.

Keywords: quality of life, caregivers, children in need of special protection, physical health, psychological health, social health, living conditions, role overload, role distress

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Authors: Eduardo Antonio Lozano Hernandez, Nancy Ramirez Alvarez, Lorena Margarita Rios Mendoza, Jose Vinicio Macias Zamora, Felix Augusto Hernandez Guzman, Jose Luis Sanchez Osorio

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Microplastics (MPs) are one of the most numerous reported wastes found in the marine ecosystem, representing one of the greatest risks for organisms that inhabit that environment due to their bioavailability. Such is the case of bivalve mollusks, since they are capable of filtering large volumes of water, which increases the risk of contamination by microplastics through the continuous exposure to these materials. This study aims to determine, quantify and characterize microplastics found in the cultured oyster Crassostrea gigas. We also analyzed if there are spatio-temporal differences in the microplastic concentration of organisms grown in two bays having quite different human population. In addition, we wanted to have an idea of the possible impact on humans via consumption of these organisms. Commercial size organisms (>6cm length; n = 15) were collected by triplicate from eight oyster farming sites in Baja California, Mexico during winter and summer. Two sites are located in Todos Santos Bay (TSB), while the other six are located in San Quintin Bay (SQB). Site selection was based on commercial concessions for oyster farming in each bay. The organisms were chemically digested with 30% KOH (w/v) and 30% H₂O₂ (v/v) to remove the organic matter and subsequently filtered using a GF/D filter. All particles considered as possible MPs were quantified according to their physical characteristics using a stereoscopic microscope. The type of synthetic polymer was determined using a FTIR-ATR microscope and using a user as well as a commercial reference library (Nicolet iN10 Thermo Scientific, Inc.) of IR spectra of plastic polymers (with a certainty ≥70% for polymers pure; ≥50% for composite polymers). Plastic microfibers were found in all the samples analyzed. However, a low incidence of MP fragments was observed in our study (approximately 9%). The synthetic polymers identified were mainly polyester and polyacrylonitrile. In addition, polyethylene, polypropylene, polystyrene, nylon, and T. elastomer. On average, the content of microplastics in organisms were higher in TSB (0.05 ± 0.01 plastic particles (pp)/g of wet weight) than found in SQB (0.02 ± 0.004 pp/g of wet weight) in the winter period. The highest concentration of MPs found in TSB coincides with the rainy season in the region, which increases the runoff from streams and wastewater discharges to the bay, as well as the larger population pressure (> 500,000 inhabitants). Otherwise, SQB is a mainly rural location, where surface runoff from streams is minimal and in addition, does not have a wastewater discharge into the bay. During the summer, no significant differences (Manne-Whitney U test; P=0.484) were observed in the concentration of MPs found in the cultured oysters of TSB and SQB, (average: 0.01 ± 0.003 pp/g and 0.01 ± 0.002 pp/g, respectively). Finally, we concluded that the consumption of oyster does not represent a risk for humans due to the low concentrations of MPs found. The concentration of MPs is influenced by the variables such as temporality, circulations dynamics of the bay and existing demographic pressure.

Keywords: FTIR-ATR, Human risk, Microplastic, Oyster

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Authors: Domingos Viegas, Miguel Almeida, Carmen Rocha, Ilda Novo, Yolanda Luna

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Forest fires along the more than 1200km of the Spanish-Portuguese border are more and more frequent, currently achieving around 2000 fire events per year. Some of these events develop to large international wildfire requiring concerted operations based on shared information between the two countries. The fire event of Valencia de Alcantara (2003) causing several fatalities and more than 13000ha burnt, is a reference example of these international events. Currently, Portugal and Spain have a specific cross-border cooperation protocol on wildfires response for a strip of about 30km (15 km for each side). It is recognized by public authorities the successfulness of this collaboration however it is also assumed that this cooperation should include more functionalities such as the development of a common risk information system for transboundary fire events. Since Portuguese and Spanish authorities use different approaches to determine the fire risk indexes inputs and different methodologies to assess the fire risk, sometimes the conjoint firefighting operations are jeopardized since the information is not harmonized and the understanding of the situation by the civil protection agents from both countries is not unique. Thus, a methodology aiming the harmonization of the fire risk calculation and perception by Portuguese and Spanish Civil protection authorities is hereby presented. The final results are presented as well. The fire risk index used in this work is the Canadian Fire Weather Index (FWI), which is based on meteorological data. The FWI is limited on its application as it does not take into account other important factors with great effect on the fire appearance and development. The combination of these factors is very complex since, besides the meteorology, it addresses several parameters of different topics, namely: sociology, topography, vegetation and soil cover. Therefore, the meaning of FWI values is different from region to region, according the specific characteristics of each region. In this work, a methodology for FWI calibration based on the number of fire occurrences and on the burnt area in the transboundary regions of Portugal and Spain, in order to assess the fire risk based on calibrated FWI values, is proposed. As previously mentioned, the cooperative firefighting operations require a common perception of the information shared. Therefore, a common classification of the fire risk for the fire events occurred in the transboundary strip is proposed with the objective of harmonizing this type of information. This work is integrated in the ECHO project SpitFire - Spanish-Portuguese Meteorological Information System for Transboundary Operations in Forest Fires, which aims the development of a web platform for the sharing of information and supporting decision tools to be used in international fire events involving Portugal and Spain.

Keywords: data harmonization, FWI, international collaboration, transboundary wildfires

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Authors: Leslie Raphael de M. Ferraz, Salvana Priscylla M. Costa, Tarcyla de A. Gomes, Giovanna Christinne R. M. Schver, Cristóvão R. da Silva, Magaly Andreza M. de Lyra, Danilo Augusto F. Fontes, Larissa A. Rolim, Amanda Carla Q. M. Vieira, Miracy M. de Albuquerque, Pedro J. Rolim-Neto

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Efavirenz (EFV) is a drug used as first-line treatment of AIDS. However, it has poor aqueous solubility and wettability, presenting problems in the gastrointestinal tract absorption and bioavailability. One of the most promising strategies to improve the solubility is the use of solid dispersions (SD). Therefore, this study aimed to characterize SD EFZ with the polymers: PVP-K30, PVPVA 64 and SOLUPLUS in order to find an optimal formulation to compose a future pharmaceutical product for AIDS therapy. Initially, Physical Mixtures (PM) and SD with the polymers were obtained containing 10, 20, 50 and 80% of drug (w/w) by the solvent method. The best formulation obtained between the SD was selected by in vitro dissolution test. Finally, the drug-carrier system chosen, in all ratios obtained, were analyzed by the following techniques: Differential Scanning Calorimetry (DSC), polarization microscopy, Scanning Electron Microscopy (SEM) and spectrophotometry of absorption in the region of infrared (IR). From the dissolution profiles of EFV, PM and SD, the values of area Under The Curve (AUC) were calculated. The data showed that the AUC of all PM is greater than the isolated EFV, this result is derived from the hydrophilic properties of the polymers thus favoring a decrease in surface tension between the drug and the dissolution medium. In adittion, this ensures an increasing of wettability of the drug. In parallel, it was found that SD whom had higher AUC values, were those who have the greatest amount of polymer (with only 10% drug). As the amount of drug increases, it was noticed that these results either decrease or are statistically similar. The AUC values of the SD using the three different polymers, followed this decreasing order: SD PVPVA 64-EFV 10% > SD PVP-K30-EFV 10% > SD Soluplus®-EFV 10%. The DSC curves of SD’s did not show the characteristic endothermic event of drug melt process, suggesting that the EFV was converted to its amorphous state. The analysis of polarized light microscopy showed significant birefringence of the PM’s, but this was not observed in films of SD’s, thus suggesting the conversion of the drug from the crystalline to the amorphous state. In electron micrographs of all PM, independently of the percentage of the drug, the crystal structure of EFV was clearly detectable. Moreover, electron micrographs of the SD with the two polymers in different ratios investigated, we observed the presence of particles with irregular size and morphology, also occurring an extensive change in the appearance of the polymer, not being possible to differentiate the two components. IR spectra of PM corresponds to the overlapping of polymer and EFV bands indicating thereby that there is no interaction between them, unlike the spectra of all SD that showed complete disappearance of the band related to the axial deformation of the NH group of EFV. Therefore, this study was able to obtain a suitable formulation to overcome the solubility limitations of the EFV, since SD PVPVA 64-EFZ 10% was chosen as the best system in delay crystallization of the prototype, reaching higher levels of super saturation.

Keywords: characterization, dissolution, Efavirenz, solid dispersions

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Authors: Ortiz M. Rocío, Rocha G. Karina, Domenech A. Gloria

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There are several tools to assess the nutritional status of the population. A main instrument commonly used to build those tools is the food composition tables (FCT). Despite the importance of FCT, there are many error sources and variability factors that can be presented on building those tables and can lead to an under or over estimation of ingest of nutrients of a population. This work identified different food composition tables used as an instrument to estimate the nutrient ingest in Ecuador.The collection of data for choosing FCT was made through key informants –self completed questionnaires-, supplemented with institutional web research. A questionnaire with general variables (origin, year of edition, etc) and methodological variables (method of elaboration, information of the table, etc) was passed to the identified FCT. Those variables were defined based on an extensive literature review. A descriptive analysis of content was performed. Ten printed tables and three databases were reported which were all indistinctly treated as food composition tables. We managed to get information from 69% of the references. Several informants referred to printed documents that were not accessible. In addition, searching the internet was not successful. Of the 9 final tables, n=8 are from Latin America, and, n= 5 of these were constructed by indirect method (collection of already published data) having as a main source of information a database from the United States department of agriculture USDA. One FCT was constructed by using direct method (bromatological analysis) and has its origin in Ecuador. The 100% of the tables made a clear distinction of the food and its method of cooking, 88% of FCT expressed values of nutrients per 100g of edible portion, 77% gave precise additional information about the use of the table, and 55% presented all the macro and micro nutrients on a detailed way. The more complete FCT were: INCAP (Central America), Composition of foods (Mexico). The more referred table was: Ecuadorian food composition table of 1965 (70%). The indirect method was used for most tables within this study. However, this method has the disadvantage that it generates less reliable food composition tables because foods show variations in composition. Therefore, a database cannot accurately predict the composition of any isolated sample of a food product.In conclusion, analyzing the pros and cons, and, despite being a FCT elaborated by using an indirect method, it is considered appropriate to work with the FCT of INCAP Central America, given the proximity to our country and a food items list that is very similar to ours. Also, it is imperative to have as a reference the table of composition for Ecuadorian food, which, although is not updated, was constructed using the direct method with Ecuadorian foods. Hence, both tables will be used to elaborate a questionnaire with the purpose of assessing the food consumption of the Ecuadorian population. In case of having disparate values, we will proceed by taking just the INCAP values because this is an updated table.

Keywords: Ecuadorian food composition tables, FCT elaborated by direct method, ingest of nutrients of Ecuadorians, Latin America food composition tables

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Authors: Marcela De Oliveira, Marina P. Da Silva, Fernando C. G. Da Rocha, Jorge M. Santos, Jaime S. Cardoso, Paulo N. Lisboa-Filho

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Multiple Sclerosis (MS) is an immune-mediated disease of the central nervous system characterized by neurodegeneration, inflammation, demyelination, and axonal loss. Magnetic resonance imaging (MRI), due to the richness in the information details provided, is the gold standard exam for diagnosis and follow-up of neurodegenerative diseases, such as MS. Brain atrophy, the gradual loss of brain volume, is quite extensive in multiple sclerosis, nearly 0.5-1.35% per year, far off the limits of normal aging. Thus, the brain volume quantification becomes an essential task for future analysis of the occurrence atrophy. The analysis of MRI has become a tedious and complex task for clinicians, who have to manually extract important information. This manual analysis is prone to errors and is time consuming due to various intra- and inter-operator variability. Nowadays, computerized methods for MRI segmentation have been extensively used to assist doctors in quantitative analyzes for disease diagnosis and monitoring. Thus, the purpose of this work was to evaluate the brain volume in MRI of MS patients. We used MRI scans with 30 slices of the five patients diagnosed with multiple sclerosis according to the McDonald criteria. The computational methods for the analysis of images were carried out in two steps: segmentation of the brain and brain volume quantification. The first image processing step was to perform brain extraction by skull stripping from the original image. In the skull stripper for MRI images of the brain, the algorithm registers a grayscale atlas image to the grayscale patient image. The associated brain mask is propagated using the registration transformation. Then this mask is eroded and used for a refined brain extraction based on level-sets (edge of the brain-skull border with dedicated expansion, curvature, and advection terms). In the second step, the brain volume quantification was performed by counting the voxels belonging to the segmentation mask and converted in cc. We observed an average brain volume of 1469.5 cc. We concluded that the automatic method applied in this work can be used for the brain extraction process and brain volume quantification in MRI. The development and use of computer programs can contribute to assist health professionals in the diagnosis and monitoring of patients with neurodegenerative diseases. In future works, we expect to implement more automated methods for the assessment of cerebral atrophy and brain lesions quantification, including machine-learning approaches. Acknowledgements: This work was supported by a grant from Brazilian agency Fundação de Amparo à Pesquisa do Estado de São Paulo (number 2019/16362-5).

Keywords: brain volume, magnetic resonance imaging, multiple sclerosis, skull stripper

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Authors: Francisco Pereira, António Augusto Vicente, Filipe Vaz, Joel Borges, Pedro Geada

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Light is a growth-limiting factor in microalgae cultivation, where factors like spectral components, intensity, and duration, often characterized by its wavelength, are well-reported to have a substantial impact on cell growth rates and, consequently, photosynthetic performance and mitigation of CO2, one of the most significant greenhouse gases (GHGs). Photobioreactors (PBRs) are commonly used to grow microalgae under controlled conditions, but they often fail to provide an even light distribution to the cultures. For this reason, there is a pressing need for innovations aiming at enhancing the efficient utilization of light. So, one potential approach to address this issue is by implementing plasmonic films, such as the localized surface plasmon resonance (LSPR). LSPR is an optical phenomenon connected to the interaction of light with metallic nanostructures. LSPR excitation is characterized by the oscillation of unbound conduction electrons of the nanoparticles coupled with the electromagnetic field from incident light. As a result of this excitation, highly energetic electrons and a strong electromagnetic field are generated. These effects lead to an amplification of light scattering, absorption, and extinction of specific wavelengths, contingent on the nature of the employed nanoparticle. Thus, microalgae might benefit from this biotechnology as it enables the selective filtration of inhibitory wavelengths and harnesses the electromagnetic fields produced, which could lead to enhancements in both biomass and metabolite productivity. This study aimed at implementing and evaluating a “plasmon-enhanced PBR”. The goal was to utilize LSPR thin films to enhance the growth and CO2 bio-fixation rate of Chlorella vulgaris. The internal/external walls of the PBRs were coated with a TiO2 matrix containing different nanoparticles (Au, Ag, and Au-Ag) in order to evaluate the impact of this approach on microalgae’s performance. Plasmonic films with distinct compositions resulted in different Chlorella vulgaris growth, ranging from 4.85 to 6.13 g.L-1. The highest cell concentrations were obtained with the metallic Ag films, demonstrating a 14% increase compared to the control condition. Moreover, it appeared to be no differences in growth between PBRs with inner and outer wall coatings. In terms of CO2 bio-fixation, distinct rates were obtained depending on the coating applied, ranging from 0.42 to 0.53 gCO2L-1d-1. Ag coating was demonstrated to be the most effective condition for carbon fixation by C. vulgaris. The impact of LSPR films on the biochemical characteristics of biomass (e.g., proteins, lipids, pigments) was analysed as well. Interestingly, Au coating yielded the most significant enhancements in protein content and total pigments, with increments of 15 % and 173 %, respectively, when compared to the PBR without any coating (control condition). Overall, the incorporation of plasmonic films in PBRs seems to have the potential to improve the performance and efficiency of microalgae cultivation, thereby representing an interesting approach to increase both biomass production and GHGs bio-mitigation.

Keywords: CO₂ bio-fixation, plasmonic effect, photobioreactor, photosynthetic microalgae

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Authors: Carlos A. C. G. Neto, Natan C. G. e Silva , Thaís de O. Costa, Luciana R. B. Gonçalves, Maria V. P. Rocha

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β-galactosidases (E.C. 3.2.1.23) are enzymes that have attracted by catalyzing the hydrolysis of lactose and in producing galacto-oligosaccharides by favoring transgalactosylation reactions. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers is a promising alternative to enhance the stability of the biocatalysts, among which polyethylenimine (PEI) stands out. PEI has certain properties, such as being a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases. Besides that, protects them from environmental variations. The use of chitosan support coated with PEI could improve the catalytic efficiency of β-galactosidase from Kluyveromyces lactis in the transgalactosylation reaction for the production of prebiotics, such as lactulose since this strain is more effective in the hydrolysis reaction. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from K. lactis immobilized on chitosan-coated with PEI, determining the immobilization parameters, its operational and thermal stability, and then to apply it in hydrolysis and transgalactolisation reactions to produce lactulose using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% (v/v) glutaraldehyde and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg protein per gram support. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey supplemented with fructose at a ratio of 1:2 lactose/fructose, totaling 200 g/L. Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6 with 0.1 mM MnCl2. The biocatalyst whose support was coated was named CHI_GLU_PEI_GAL, and the one that was not coated was named CHI_GLU_GAL. The coating of the support with PEI considerably improved the parameters of immobilization. The immobilization yield increased from 56.53% to 97.45%, biocatalyst activity from 38.93 U/g to 95.26 U/g and the efficiency from 3.51% to 6.0% for uncoated and coated support, respectively. The biocatalyst CHI_GLU_PEI_GAL was better than CHI_GLU_GAL in the hydrolysis of lactose and production of lactulose, converting 97.05% of lactose at 5 min of reaction and producing 7.60 g/L lactulose in the same time interval. QUI_GLU_PEI_GAL biocatalyst was stable in the hydrolysis reactions of lactose during the 10 cycles evaluated, converting 73.45% lactose even after the tenth cycle, and in the lactulose production was stable until the fifth cycle evaluated, producing 10.95 g/L lactulose. However, the thermal stability of CHI_GLU_GAL biocatalyst was superior, with a half-life time 6 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (CHI_GLU_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as, the catalytic action of the enzyme. Besides that, this process can be economically viable due to the use of an industrial residue as a substrate.

Keywords: β-galactosidase, immobilization, kluyveromyces lactis, lactulose, polyethylenimine, transgalactosylation reaction, whey

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Authors: Guido Lorenzi, Massimo Santarelli, Carlos Augusto Santos Silva

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The growth of non-dispatchable renewable energy sources in the European electricity generation mix is promoting the research of technically feasible and cost-effective solutions to make use of the excess energy, produced when the demand is low. The increasing intermittent renewable capacity is becoming a challenge to face especially in Europe, where some countries have shares of wind and solar on the total electricity produced in 2015 higher than 20%, with Denmark around 40%. However, other consumption sectors (mainly transportation) are still considerably relying on fossil fuels, with a slow transition to other forms of energy. Among the opportunities for different mobility concepts, electric (EV) and biofuel-powered vehicles (BPV) are the options that currently appear more promising. The EVs are targeting mainly the light duty users because of their zero (Full electric) or reduced (Hybrid) local emissions, while the BPVs encourage the use of alternative resources with the same technologies (thermal engines) used so far. The batteries which are applied to EVs are based on ions of Lithium because of their overall good performance in energy density, safety, cost and temperature performance. Biofuels, instead, can be various and the major difference is in their physical state (liquid or gaseous). In this study gaseous biofuels are considered and, more specifically, Synthetic Natural Gas (SNG) produced through a process of Power-to-Gas consisting in an electrochemical upgrade (with Solid Oxide Electrolyzers) of biogas with CO2 recycling. The latter process combines a first stage of electrolysis, where syngas is produced, and a second stage of methanation in which the product gas is turned into methane and then made available for consumption. A techno-economic comparison between the two alternatives is possible, but it does not capture all the different aspects involved in the two routes for the promotion of a more sustainable mobility. For this reason, a more comprehensive methodology, i.e. Life Cycle Assessment, is adopted to describe the environmental implications of using excess electricity (directly or indirectly) for new vehicle fleets. The functional unit of the study is 1 km and the two options are compared in terms of overall CO2 emissions, both considering Cradle to Gate and Cradle to Grave boundaries. Showing how production and disposal of materials affect the environmental performance of the analyzed routes is useful to broaden the perspective on the impacts that different technologies produce, in addition to what is emitted during the operational life. In particular, this applies to batteries for which the decommissioning phase has a larger impact on the environmental balance compared to electrolyzers. The lower (more than one order of magnitude) energy density of Li-ion batteries compared to SNG implies that for the same amount of energy used, more material resources are needed to obtain the same effect. The comparison is performed in an energy system that simulates the Western European one, in order to assess which of the two solutions is more suitable to lead the de-fossilization of the transport sector with the least resource depletion and the mildest consequences for the ecosystem.

Keywords: electrical energy storage, electric vehicles, power-to-gas, life cycle assessment

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Authors: Carlos A. C. G. Neto, Natan C. G. Silva, Thais O. Costa, Luciana R. B. Goncalves, Maria v. P. Rocha

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Galactosidases are enzymes responsible for catalyzing lactose hydrolysis reactions and also favoring transgalactosylation reactions for the production of prebiotics, among which lactulose stands out. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers in immobilization processes is a promising alternative in order to extend the useful life of the biocatalysts, for example, the coating with polyethyleneimine (PEI). PEI is a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases and also protects it from environmental variations, for example, pH and temperature. In addition, it can substantially improve the immobilization parameters and also the efficiency of enzymatic reactions. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from Kluyveromyces lactis immobilized on PEI coated agarose, determining the immobilization parameters, its operational and thermal stability, and then to apply it in the hydrolysis of lactose and synthesis of lactulose, using whey as a substrate. This immobilization strategy was chosen in order to improve the catalytic efficiency of the enzyme in the transgalactosylation reaction for the production of prebiotics, and there are few studies with β-galactosidase from this strain. The immobilization of β-galactosidase in agarose previously functionalized with 48% (w/v) glycidol and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg/g of protein. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey (66.7 g/L of lactose) supplemented with 133.3 g/L fructose at a ratio of 1:2 (lactose/fructose). Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6, with 0.1 mM MnCl2. The biocatalysts whose supports were coated were named AGA_GLY_PEI_GAL, and those that were not coated were named AGA_GLY_GAL. The coating of the support with PEI considerably improved immobilization yield (2.6-fold), the biocatalyst activity (1.4-fold), and efficiency (2.2-fold). The biocatalyst AGA_GLY_PEI_GAL was better than AGA_GLY_GAL in hydrolysis and transgalactosylation reactions, converting 88.92% of lactose at 5 min of reaction and obtaining a residual concentration of 5.24 g/L. Besides that, it was produced 13.90 g/L lactulose in the same time interval. AGA_GLY_PEI_GAL biocatalyst was stable during the 10 cycles evaluated, converting approximately 80% of lactose and producing 10.95 g/L of lactulose even after the tenth cycle. However, the thermal stability of AGA_GLY_GAL biocatalyst was superior, with a half-life time 5 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (AGA_GLY_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as the enzyme, catalyzed reactions. In addition, the use of whey as a raw material for lactulose production has proved to be an industrially advantageous alternative.

Keywords: β-galactosidase, immobilization, lactulose, polyethylenimine, whey

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Authors: Zélia Maria Da Costa Ludwig, Maria José Valenzuela Bell, Geraldo Henriques Da Silva, Thales Alves Faraco, Victor Rocha Da Silva, Daniel Rotmeister Teixeira, Vírgilio De Carvalho Dos Anjos, Valdemir Ludwig

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Advances in telecommunications grow with the development of optical amplifiers based on rare earth ions. The focus has been concentrated in silicate glasses although their amplified spontaneous emission is limited to a few tens of nanometers (~ 40nm). Recently, phosphate glasses have received great attention due to their potential application in optical data transmission, detection, sensors and laser detector, waveguide and optical fibers, besides its excellent physical properties such as high thermal expansion coefficients and low melting temperature. Compared with the silica glasses, phosphate glasses provide different optical properties such as, large transmission window of infrared, and good density. Research on the improvement of physical and chemical durability of phosphate glass by addition of heavy metals oxides in P2O5 has been performed. The addition of Na2O further improves the solubility of rare earths, while increasing the Al2O3 links in the P2O5 tetrahedral results in increased durability and aqueous transition temperature and a decrease of the coefficient of thermal expansion. This work describes the structural and spectroscopic characterization of a phosphate glass matrix doped with different Er (Erbium) concentrations. The phosphate glasses containing Er3+ ions have been prepared by melt technique. A study of the optical absorption, luminescence and lifetime was conducted in order to characterize the infrared emission of Er3+ ions at 1540 nm, due to the radiative transition 4I13/2 → 4I15/2. Our results indicate that the present glass is a quite good matrix for Er3+ ions, and the quantum efficiency of the 1540 nm emission was high. A quenching mechanism for the mentioned luminescence was not observed up to 2,0 mol% of Er concentration. The Judd-Ofelt parameters, radiative lifetime and quantum efficiency have been determined in order to evaluate the potential of Er3+ ions in new phosphate glass. The parameters follow the trend as Ω2 > Ω4 > Ω6. It is well known that the parameter Ω2 is an indication of the dominant covalent nature and/or structural changes in the vicinity of the ion (short range effects), while Ω4 and Ω6 intensity parameters are long range parameters that can be related to the bulk properties such as viscosity and rigidity of the glass. From the PL measurements, no red or green upconversion was measured when pumping the samples with laser excitation at 980 nm. As future prospects: Synthesize this glass system with silver in order to determine the influence of silver nanoparticles on the Er3+ ions.

Keywords: phosphate glass, erbium, luminescence, glass system

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Authors: Marcelo Lopes Dias Kolling, Felipe Ferreira Laranjeira, Guilherme Augusto Hettwer, Pedro Salomão Piccinini, Marwan Masri, Carlos Oscar Uebel

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Introduction: Injectable fillers are among the most common nonsurgical cosmetic procedures, with significant growth yearly. Knowledge of rheological and mechanical characteristics of fillers, facial anatomy, and injection technique is essential for safety. Concepts such as the use of cannula versus needle, aspiration before injection, and facial danger zones have been well discussed. In case of an accidental intravascular puncture, the pressure inside the vessel may not be sufficient to push blood into the syringe due to the characteristics of the filler product; this is especially true for calcium hydroxyapatite (CaHA) or hyaluronic acid (HA) fillers with high G’. Since viscoelastic properties of normal saline are much lower than those of fillers, aspiration with saline prior to filler injection may decrease the risk of a false negative aspiration and subsequent catastrophic effects. We discuss a technique to add an additional safety step to the procedure with saline aspiration prior to injection, a ‘’reverse Seldinger’’ technique for intravascular access, which we term SANIT: Saline Aspiration Negative Intravascular Test. Objectives: To demonstrate the author’s (PSP) technique which adds an additional safety step to the process of filler injection, with both CaHA and HA, in order to decrease the risk of intravascular injection. Materials and Methods: Normal skin cleansing and topical anesthesia with prilocaine/lidocaine cream are performed; the facial subunits to be treated are marked. A 3mL Luer lock syringe is filled with 2mL of 0.9% normal saline and a 27G needle, which is turned one half rotation. When a cannula is to be used, the Luer lock syringe is attached to a 27G 4cm single hole disposable cannula. After skin puncture, the 3mL syringe is advanced with the plunger pulled back (negative pressure). Progress is made to the desired depth, all the while aspirating. Once the desired location of filler injection is reached, the syringe is exchanged for the syringe containing a filler, securely grabbing the hub of the needle and taking care to not dislodge the needle tip. Prior to this, we remove 0.1mL of filler to allow for space inside the syringe for aspiration. We again aspirate and inject retrograde. SANIT is especially useful for CaHA, since the G’ is much higher than HA, and thus reflux of blood into the syringe is less likely to occur. Results: The technique has been used safely for the past two years with no adverse events; the increase in cost is negligible (only the cost of 2mL of normal saline). Over 100 patients (over 300 syringes) have been treated with this technique. The risk of accidental intravascular puncture has been calculated to be between 1:6410 to 1:40882 syringes among expert injectors; however, the consequences of intravascular injection can be catastrophic even with board-certified physicians. Conclusions: While the risk of intravascular filler injection is low, the consequences can be disastrous. We believe that adding the SANIT technique can help further mitigate risk with no significant untoward effects and could be considered by all performing injectable fillers. Further follow-up is ongoing.

Keywords: injectable fillers, safety, saline aspiration, injectable filler complications, hyaluronic acid, calcium hydroxyapatite

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Authors: Rayanne Branco Dos Santos Lima, Lorena Pinheiro Barbosa, Kamila Ferreira Lima, Victor Manuel Tegoma Ruiz, Monyka Brito Lima Dos Santos, Maria Wendiane Gueiros Gaspar, Luzia Camila Coelho Ferreira, Leandro Cardozo Dos Santos Brito, Deyse Maria Alves Rocha

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Introduction: The lack of regulation of the baby's sleep-wake pattern in the first years of life affects the health of thousands of women. Maternal sleep deprivation can trigger or aggravate psychosomatic problems such as depression, anxiety and stress that can directly influence maternal safety, with consequences for the baby's and mother's sleep. Such conditions can affect the family's quality of life and child development. Objective: To correlate maternal security with maternal state anxiety scores and the mother's and baby's total sleep time. Method: Cross-sectional study carried out with 96 mothers of babies aged 10 to 24 months, accompanied by nursing professionals linked to a Federal University in Northeast Brazil. Study variables were maternal security, maternal state anxiety scores, infant latency and sleep time, and total nocturnal sleep time of mother and infant. Maternal safety was calculated using a four-point Likert scale (1=not at all safe, 2=somewhat safe, 3=very safe, 4=completely safe). Maternal anxiety was measured by State-Trait Anxiety Inventory, state-anxiety subscale whose scores vary from 20 to 80 points, and the higher the score, the higher the anxiety levels. Scores below 33 are considered mild; from 33 to 49, moderate and above 49, high. As for the total nocturnal sleep time, values between 7-9 hours of sleep were considered adequate for mothers, and values between 9-12 hours for the baby, according to the guidelines of the National Sleep Foundation. For the sleep latency time, a time equal to or less than 20 min was considered adequate. It is noteworthy that the latency time and the time of night sleep of the mother and the baby were obtained by the mother's subjective report. To correlate the data, Spearman's correlation was used in the statistical package R version 3.6.3. Results: 96 women and babies participated, aged 22 to 38 years (mean 30.8) and 10 to 24 months (mean 14.7), respectively. The average of maternal security was 2.89 (unsafe); Mean maternal state anxiety scores were 43.75 (moderate anxiety). The babies' average sleep latency time was 39.6 min (>20 min). The mean sleep times of the mother and baby were, respectively, 6h and 42min and 8h and 19min, both less than the recommended nocturnal sleep time. Maternal security was positively correlated with maternal state anxiety scores (rh=266, p=0.009) and negatively correlated with infant sleep latency (rh= -0.30. P=0.003). Baby sleep time was positively correlated with maternal sleep time. (rh 0.46, p<0.001). Conclusion: The more secure the mothers considered themselves, the higher the anxiety scores and the shorter the baby's sleep latency. Also, the longer the baby sleeps, the longer the mother sleeps. Thus, interventions are needed to promote the quality and efficiency of sleep for both mother and baby.

Keywords: sleep, anxiety, infant, mother-child relations

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Authors: Gabriela R. Silveira, Claudia S. Rocha, Lais S. Vitti, Jeane L. Borges, Helen B. Durgante

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Emerging adulthood occurs after adolescence in a period that maybe be marked by experimentation, identity reconfigurations, labor life demands, and insertion in the work environment, which tends to generate stress and emotional instability. Health promotion programs for the development of strengths and virtues, based on Positive Psychology, for emerging adults are sparse in Brazil. The aim of this study was to evaluate the preliminary effects of an online multi-component Positive Psychology program for the health promotion of emerging adults based on Cognitive Behavioural Therapy and Positive Psychology. The program included six online (synchronous) weekly group sessions of approximately two hours each and homework (asynchronous) activities. The themes worked were Values and self-care/Prudence, Optimism, Empathy, Gratitude, Forgiveness, and Meaning of life and work. This study presents data from a longitudinal, pre-experimental design with pre (T1) and post-test (T2) evaluation in the intervention group. 47 individuals aged between 19-30 years old participated, mean age of 24.53 years (SD=3.13), 37 females (78.7%). 42 (89.4%) self-defined as heterosexual, four (8.5%) as homosexual, and one (2.5%) as bisexual. 33 (70.2%) had incomplete higher education, four (8.5%) completed higher education, and seven (14.9%) had a graduate level of education. 27 participants worked (57.4%), out of which 25 were health workers (53.2%). 14 (29.8%) were caregivers, 27 (57.4%) had a spiritual belief, 36 (76.6%) had access to leisure, and 38 (80.9%) had perceived social support. The instruments used were a sociodemographic questionnaire, the 10-item Perceived Stress Scale, and the 12-item General Health Questionnaire. The program was advertised on social networks and interested participants filled out the Consent Form and the evaluation protocol at T1 and T2 via Google Docs form. The main research was approved (CEP n.1,899,368; 4,143,219; CAAE: 61997516.5.0000.5334) and complied with sanitary and Ethics criteria in research with human beings. Wilcoxon statistics revealed significant improvements in indicators of perceived stress between T1 (X=22.21, SD=6.79) and T2 (X=15.10, SD=5.82); (Z=-4.353; p=0.001) as well as depression and anxiety symptoms (T1:X=26.72, SD=8.84; T2: X=19.23, SD=4.68); (Z=-3.945, p=0.001) of the emerging adults after their participation in the programme. The programme has an innovative character not only for presenting an online Positive Psychology approach but also for being based on an intervention developed, evaluated, and manualized in Brazil. By focusing on emerging adults, this study contributes to advancing research on a relatively new field in developmental studies. As a limitation, this is a pre-experimental and pilot study, requiring an increase in sample size for greater statistical robustness, also qualitative data analysis is crucial for methodological complementarity. The importance of investing efforts to accompany this age group and provide advances in longitudinal research in the area of health promotion and disease prevention is highlighted.

Keywords: emerging adults, disease prevention, health promotion, online program

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Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

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Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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Authors: Delsy Salinas, Andreia Garcês, Augusto Silva, Paula Brilhante Simões

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Urinary tract infection (UTI) is a common disease affecting dogs and cats in both community and hospital environment. Bacteria is the most frequent agent isolated, fewer than 1% of infections are due to parasitic, fungal, or viral agents. Common symptoms and laboratory abnormalities includeabdominal pain, pyrexia, renomegaly, and neutrophilia with left shift. A rapid and precise identification of the bacterial agent is still a challenge in veterinarian laboratories. Therefore, this cross-sectional study aims to describe bacterial colony patterns of urine samples by using chromID™ CPS® EliteAgar (BioMérieux, France) from canine and feline specimens submitted to a veterinary laboratory in Portugal (INNO Veterinary Laboratory, Braga)from January to March2022. All urine samples were cultivated in CPS Elite Agar with calibrated 1 µL inoculating loop and incubated at 37ºC for 18-24h. Color,size, and shape (regular or irregular outline)were recorded for all samples. All colonies were classified as Gram-negative or Gram-positive bacteriausing Gram stain (PREVI® Color BioMérieux, France) and determined if they were pure colonies. Identification of bacteria species was performed using GP and GN cards inVitek 2® Compact(BioMérieux, France). A total of 256/1003 submitted urine samples presented bacterial growth, from which 172 isolates were included in this study. The sample’s population included 111 dogs (n=45 males and n=66 females) and 61 cats (n=35 males and n=26 females). The most frequent isolated bacteria wasEscherichia coli (44,7%), followed by Proteus mirabilis (13,4%). All Escherichia coli isolates presented red to burgundy colonies, a colony diameter between 2 to 6 mm, and regular or irregular outlines. Similarly, 100% of Proteus mirabilis isolates were dark yellow colonies with a diffuse pigment and the same size and shape as Escherichia coli. White and pink pale colonies where Staphylococcus species exclusively and S. pseudintermedius was the most frequent (8,2 %). Cian to blue colonies were mostly Enterococcusspp. (8,2%) and Streptococcus spp. (4,6%). Beige to brown colonies were Pseudomonas aeruginosa (2,9%) and Citrobacter spp. (1,2%).Klebsiella spp.,Serratia spp. and Enterobacter spp were green colonies. All Gram-positive isolates were 1 to 2 mm diameter long and had a regular outline, meanwhile, Gram-negative rods presented variable patterns. This results showed that theprevalence of E coli and P. mirabilis as uropathogenic agents follows the same trends in Europe as previously described in other studies. Both agents presented a particular color pattern in CPS Elite Agar to identify them without needing complementary tests. No other bacteria genus could be correlated strongly to a specific color pattern, and similar results have been observed instudies using human’s samples. Chromogenic media shows a great advantage for common urine bacteria isolation than traditional COS, McConkey, and CLEDAgar mediums in a routine context, especially when mixed fermentative Gram-negative agents grow simultaneously. In addition, CPS Elite Agar is versatile for Artificial Intelligent Reading Plates Systems. Routine veterinarian laboratories could use CPS Elite Agar for a rapid screening for bacteria identification,mainlyE coli and P.mirabilis, saving 6h to 10h of automatized identification.

Keywords: cats, CPS elite agar, dogs, urine pathogens

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Authors: Gousilin Leandra Rocha Da Silva, Stéfani T. A. Dantas, Bruna F. Rossi, Erika R. Bonsaglia, Ivana G. Castilho, Terue Sadatsune, Ary Fernandes Júnior, Vera l. M. Rall

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Kidney failure causes decreased diuresis and accumulation of nitrogenous substances in the body. To increase patient survival, hemodialysis is used as a partial substitute for renal function. However, contamination of the water used in this treatment, causing bacteremia in patients, is a worldwide concern. The Burkholderia cepacia complex (Bcc), a group of bacteria with more than 20 species, is frequently isolated from hemodialysis water samples and comprises opportunistic bacteria, affecting immunosuppressed patients, due to its wide variety of virulence factors, in addition to innate resistance to several antimicrobial agents, contributing to the permanence in the hospital environment and to the pathogenesis in the host. The objective of the present work was to characterize molecularly and phenotypically Bcc isolates collected from the water and dialysate of the Hemodialysis Unit and from the blood of patients at a Public Hospital in Botucatu, São Paulo, Brazil, between 2019 and 2021. We used 33 Bcc isolates, previously obtained from blood cultures from patients with bacteremia undergoing hemodialysis treatment (2019-2021) and 24 isolates obtained from water and dialysate samples in a Hemodialysis Unit (same period). The recA gene was sequenced to identify the specific species among the Bcc group. All isolates were tested for the presence of some genes that encode virulence factors such as cblA, esmR, zmpA and zmpB. Considering the epidemiology of the outbreak, the Bcc isolates were molecularly characterized by Multi Locus Sequence Type (MLST) and by pulsed-field gel electrophoresis (PFGE). The verification and quantification of biofilm in a polystyrene microplate were performed by submitting the isolates to different incubation temperatures (20°C, average water temperature and 35°C, optimal temperature for group growth). The antibiogram was performed with disc diffusion tests on agar, using discs impregnated with cefepime (30µg), ceftazidime (30µg), ciprofloxacin (5µg), gentamicin (10µg), imipenem (10µg), amikacin 30µg), sulfametazol/trimethoprim (23.75/1.25µg) and ampicillin/sulbactam (10/10µg). The presence of ZmpB was identified in all isolates, while ZmpA was observed in 96.5% of the isolates, while none of them presented the cblA and esmR genes. The antibiogram of the 33 human isolates indicated that all were resistant to gentamicin, colistin, ampicillin/sulbactam and imipenem. 16 (48.5%) isolates were resistant to amikacin and lower rates of resistance were observed for meropenem, ceftazidime, cefepime, ciprofloxacin and piperacycline/tazobactam (6.1%). All isolates were sensitive to sulfametazol/trimethoprim, levofloxacin and tigecycline. As for the water isolates, resistance was observed only to gentamicin (34.8%) and imipenem (17.4%). According to PFGE results, all isolates obtained from humans and water belonged to the same pulsotype (1), which was identified by recA sequencing as B. cepacia¸, belonging to sequence type ST-767. By observing a single pulse type over three years, one can observe the persistence of this isolate in the pipeline, contaminating patients undergoing hemodialysis, despite the routine disinfection of water with peracetic acid. This persistence is probably due to the production of biofilm, which protects bacteria from disinfectants and, making this scenario more critical, several isolates proved to be multidrug-resistant (resistance to at least three groups of antimicrobials), turning the patient care even more difficult.

Keywords: hemodialysis, burkholderia cepacia, PFGE, MLST, multi drug resistance

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