Generation of a novel three-dimensional scaffold-based model of the bovine endometrium.

Bovine in vitro endometrial models that resemble tissue function in vivo are needed to study infertility, long-term uterine alterations induced by pathogens and impact of endocrine disruptor chemicals on reproductive function and other reproductive system complications that cause high economic losses in livestock species. The present study aimed to generate an innovative, reproducible, and functional 3D scaffold-based model of the bovine endometrium structurally robust for long term-culture. We developed a multicellular model containing both endometrial epithelial and stromal cells. Epithelial cells organized to form a luminal-like epithelial layer on the surface of the scaffold. Stromal cells produced their own extracellular matrix forming a stable subepithelial compartment that physiologically resembles the normal endometrium. Both cell types released prostaglandin E2 and prostaglandin F2α following a treatment with oxytocin and arachidonic acid. Additionally signal pathways mediating oxytocin and arachidonic acid stimulation of prostaglandin synthesis were analyzed by real time PCR (RT-PCR). Oxytocin receptor (OXTR), prostaglandin E2 receptor 2 (EP2), prostaglandin E2 receptor 4 (EP4), prostaglandin F receptor (PTGFR), prostaglandin E synthase (PTGES), PGF-synthase (PGFS) and prostaglandin-endoperoxide synthase 2 (COX-2) expression was detected in both control and treatment groups, however, only significant changes in abundance of OXTR mRNA transcripts were found. The results obtained by this study are a step forward in bovine in vitro culture technology. This 3D scaffold-based model provides a platform to study regulatory mechanisms involved in endometrial physiology and can set the basis for a broader tool for designing and testing novel therapeutic strategies for recurrent uterine pathologies.

Silver, copper and copper oxide nanoparticles in the fight against human viruses: progress and perspectives.

The rapid development of nanomedicine has created a high demand for silver, copper and copper oxide nanoparticles. Due to their high reactivity and potent antimicrobial activity, silver and copper-based nanomaterials have been playing an important role in the search for new alternatives for the treatment of several issues of concern, such as pathologies caused by bacteria and viruses. Viral diseases are a significant and constant threat to public health. The most recent example is the pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this context, the object of the present review is to highlight recent progress in the biomedical uses of these metal nanoparticles for the treatment and prevention of human viral infections. We discuss the antiviral activity of AgNPs and Cu-based NPs, including their actions against SARS-CoV-2. We also discuss the toxicity, biodistribution and excretion of AgNPs and CuNPs, along with their uses in medical devices or on inert surfaces to avoid viral dissemination by fomites. The challenges and limitations of the biomedical use of these nanoparticles are presented.

Silver nanoparticles: Toxicity in model organisms as an overview of its hazard for human health and the environment.

Silver nanoparticles (AgNPs) have attracted remarkable attention due to their powerful antimicrobial action as well as their particular physicochemical properties. This has led to their application in a wide variety of products with promising results. However, their interaction with the environment and toxicity in live terrestrial or aquatic organisms is still a matter of intense debate. More detailed knowledge is still required about the toxicity of AgNPs, their possible uptake mechanisms and their adverse effects in live organisms. Several studies have reported the interactions and potential negative effects of AgNPs in different organisms. In this review, we report and discuss the current state of the art and perspectives for the impact of AgNPs on different organisms present in the environment. Recent progress in interpreting uptake, translocation and accumulation mechanisms in different organisms and/or living animals are discussed, as well as the toxicity of AgNPs and possible tolerance mechanisms in live organisms to cope with their deleterious effects. Finally, we discuss the challenges of accurate physicochemical characterization of AgNPs and their ecotoxicity in environmentally realistic conditions such as soil and water media.

Mobbing perception among Intensive Care Unit nurses. / Percepción del mobbing entre el personal de enfermería de la Unidad de Cuidados Intensivos.

AIM: to determine the perception of intensive care unit nursing staff on mobbing. METHOD: qualitative approach study, Grounded Theory was used, 12intensive care unit nurses of 2public hospitals in our country during December 2017. RESULTS: female sex predominated with an average age of 41.33 years old, mostly married, on night shift and trained a nursing technicians; 4categories emerged: general knowledge about mobbing, the origin of mobbing and its main actors, experiences of mobbing as a victim and as a spectator and the implications of mobbing in working life. DISCUSSION: issues of workplace harassment are sensitive for most health workers, since they deal with private situations and lack of support from superiors when they have been victims of harassment. The evidence shows that one of the reasons why mobbing can be perceived in different ways is because little is known about the real concept, it can be associated with multiple forms of violence and there is heterogeneity in the use of the term. CONCLUSION: the majority of intensive care unit nursing staff have been victims and witnesses of mobbing behaviour, with negative repercussions on their job satisfaction and performance; It is also the cause of constant staff turnover.

Combined pollution of copper nanoparticles and atrazine in soil: Effects on dissipation of the pesticide and on microbiological community profiles.

Copper nanoparticles (NCu) have been proposed as an antimicrobial agent in agriculture. Therefore, NCu may interact with numerous pollutants including pesticides. Little is known about the combined effects of NCu and pesticides in soil. This study aimed at assessing the impact of NCu combined with the herbicide atrazine (ATZ) on soil. We focused on assessing the adsorption and dissipation of ATZ in the presence of NCu and the changes in microbial community profiles. First, ATZ adsorption isotherms (described using the Freundlich equation) were evaluated. After that, soil samples were spiked with NCu (40-60 nm) at 0.05 and 0.15% w/w and ATZ (3 mg a.i kg-1) and incubated for 30 days. The results showed that ATZ adsorption is favored by the presence of NCu. On the other hand, NCu at 0.15% w/w caused a significant decrease in ATZ dissipation, increasing its half-life from 6 to 37 days. Microbial community profiles (bacteria, fungi and nitrifying bacteria) remained relatively stable throughout the evaluated period. Therefore, our findings suggest that NCu can increase the persistence of ATZ in soil, which may be mostly associated to physical-chemical interaction with soil particles more than a microbial impact.

Pesticide dissipation and microbial community changes in a biopurification system: influence of the rhizosphere.

The dissipation of atrazine, chlorpyrifos and iprodione in a biopurification system and changes in the microbial and some biological parameters influenced by the rhizosphere of Lolium perenne were studied in a column system packed with an organic biomixture. Three column depths were analyzed for residual pesticides, peroxidase, fluorescein diacetate activity and microbial communities. Fungal colonization was analyzed by confocal laser scanning microscopy to assess the extent of its proliferation in wheat straw. The L. perenne rhizosphere enhanced pesticide dissipation and negligible pesticide residues were detected at 20-30 cm column depth. Atrazine, chlorpyrifos and iprodione removal was 82, 89 and 74% respectively in the first 10 cm depth for columns with vegetal cover. The presence of L. perenne in contaminated columns stimulated peroxidase activity in all three column depth sections. Fluorescein diacetate activity decreased over time in all column sections with the highest values in biomixtures with vegetal cover. Microbial communities, analyzed by PCR-DGGE, were not affected by the pesticide mixture application, presenting high values of similarity (>65%) with and without vegetal cover. Microbial abundance of Actinobacteria varied according to treatment and no clear link was observed. However, bacterial abundance increased over time and was similar with and without vegetal cover. On the other hand, fungal abundance decreased in all sections of columns after 40 days, but an increase was observed in response to pesticide application. Fungal colonization and straw degradation during pesticide dissipation were verified by monitoring the lignin autofluorescence loss.

Production of ligninolytic enzymes and some diffusible antifungal compounds by white-rot fungi using potato solid wastes as the sole nutrient source.

AIMS: The aim of this study was to evaluate the synthesis of ligninolytic enzymes and some diffusible antifungal compounds by white-rot fungi (WRF) using peels or discarded potato as the sole nutrient source. METHODS AND RESULTS: The strain Trametes hirsuta Ru-513 highlighted for its laccase activity (595 ± 33 U l-1 ), which is able to decolourize 87% of an anthraquinone dye using potato peels as the sole nutritional support. A native polyacrylamide gel of laccase proteins showed the presence of two isoenzymes, corresponding to proteins of 56 and 67 kDa, which were detected by SDS-PAGE. The antifungal activity of ethyl acetate extracts was evaluated by the agar diffusion method, where Anthracophyllum discolor Sp4 and Inonotus sp. Sp2 showed the highest inhibition zones of Mucor miehei. The fungal extracts also inhibited Fusarium oxysporum and Botrytis cinerea growth, with inhibition zones of up to 18 mm. The extract with the highest antifungal activity, from A. discolor Sp4 grown in discarded potato medium, was analysed using a gas chromatograph coupled to a mass spectrometer. Among the identified compounds, chlorinated aromatic compounds and veratryl alcohol were the most abundant compounds. CONCLUSIONS: The results revealed the relevance of potato waste valorization for the sustainable production of ligninolytic enzymes and antifungal compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the synthesis of ligninolytic enzymes and diffusible antifungal compounds by WRF using potato wastes as the sole nutrient source and suggests a relationship between the enzymatic activity and the synthesis of antifungal compounds. These compounds and the synthesis of halogen compounds by WRF using agro-industrial wastes have been poorly studied before.

Why teaching empathy is important for the medical degree. / ¿Por qué es importante la enseñanza de la empatía en el Grado de Medicina?

Empathy is a basic skill in the exercise of medicine and increases patient and physician satisfaction and improves clinical results. However, the teaching of empathy is poorly covered in the teaching plans. A number of studies have observed a reduction in empathy during the final training courses. The reasons for this decline include, the students' excessive academic workload, the prioritisation of acquiring medical expertise over humanistic knowledge, the patient load in hospitals and health centres and the physicians' need to distance themselves from their patients. Nevertheless, intervention studies through simulation with standardised patients have shown an increase in empathy in students, which can be evaluated through the Jefferson scales: JSE-S and JSPPPE. The teaching of empathy to medical students is an important commitment in the curricular programs of medical schools.

Bioaugmentation and rhizosphere-assisted biodegradation as strategies for optimization of the dissipation capacity of biobeds.

Biobeds are on-farm biodepuration systems whose efficiency rely on their high pesticide biodegradation capacity. We evaluated two optimization strategies, bioaugmentation and/or rhizosphere-assisted biodegradation, to maximize the dissipation capacity of biobeds. Iprodione was used as a model pesticide. Its dissipation and metabolism was determined in a biobed packing material inoculated with an iprodione-degrading Arthrobacter strain C1 (bioaugmentation, treatments B+C1) and/or seeded with ryegrass (rhizosphere-assisted biodegradation, treatments B+P). The impact of those strategies on the activity and composition of the microbial community was determined. Bioaugmentation accelerated the dissipation of iprodione which was further enhanced in the bioaugmented, rhizosphere-assisted treatment (treatment B+P+C1, Half-life (DT50) = 3.4 d), compared to the non-bioaugmented, non rhizosphere-assisted control (DT50 = 9.5 d, treatment B). Bioaugmentation resulted in the earlier formation of intermediate formation of metabolites I (3,5-dichlorophenyl-carboxamide), II (3,5-dichlorophenylurea acetate) and 3,5-dichloroaniline (3,5-DCA). The latter was further dissipated by the indigenous microbial community. Acid phosphatase (AP) and ß-glucosidase (GLU) were temporarily stimulated in rhizosphere-assisted treatments, whereas a stimulation of the fluorescein diacetate (FDA) hydrolytic activity in the bioaugmented treatments coincided with the hydrolysis of iprodione. q-PCR showed that changes in the abundance of alpha-proteobacteria and firmicutes was driven by the presence of rhizosphere while bioaugmentation had no significant effect.

Immobilization of the white-rot fungus Anthracophyllum discolor to degrade the herbicide atrazine.

Herbicides cause environmental concerns because they are toxic and accumulate in the environment, food products and water supplies. There is a need to develop safe, efficient and economical methods to remove them from the environment, often by biodegradation. Atrazine is such herbicide. White-rot fungi have the ability to degrade herbicides of potential utility. This study formulated a novel pelletized support to immobilize the white-rot fungus Anthracophyllum discolor to improve its capability to degrade the atrazine using a biopurification system (BS). Different proportions of sawdust, starch, corn meal and flaxseed were used to generate three pelletized supports (F1, F2 and F3). In addition, immobilization with coated and uncoated pelletized supports (CPS and UPS, respectively) was assessed. UPS-F1 was determined as the most effective system as it provided high level of manganese peroxidase activity and fungal viability. The half-life (t1/2) of atrazine decreased from 14 to 6 days for the control and inoculated samples respectively. Inoculation with immobilized A. discolor produced an increase in the fungal taxa assessed by DGGE and on phenoloxidase activity determined. The treatment improves atrazine degradation and reduces migration to surface and groundwater.

Increased diazinon hydrolysis to 2-isopropyl-6-methyl-4-pyrimidinol in liquid medium by a specific Streptomyces mixed culture.

Actinobacteria identified as Streptomyces spp. were evaluated for their ability to remove diazinon as the only carbon source from a liquid medium. Single cultures of Streptomyces strains were exposed to diazinon at a concentration of 50 mg L(-1). After 96 h incubation, six of the eight cultures grew and five strains showed an increase in their total protein concentrations and changes in their protein profile. Up to 32% of the diazinon was removed by the single Streptomyces cultures. A compatibility assay showed that the different Streptomyces species were not antagonistic. Twenty-six mixed cultures were then prepared. Diazinon removal was increased when mixed cultures were used, and maximum diazinon removal of 62% was observed when the Streptomyces spp. strains AC5, AC9, GA11 and ISP13 were mixed; this was defined as the selected mixed culture (SMC). Diazinon removal was positively influenced by the addition of glucose into the liquid medium. Our study showed a diazinon degradation rate of 0.025 h(-1), half-life of 28 h(-1) and 2-isopropyl-6-methyl-4-pyrimidinol (IMHP) production of 0.143 mg L h(-1). Rapid diazinon hydrolysis to IMHP was associated with a decrease in the pH of the medium as a consequence of microbial glucose metabolism and organic acid exudation. Moreover, the SMC of Streptomyces was able to remove IMHP. This work constitutes a new, if not the only, report on diazinon degradation by mixed cultures of Streptomyces spp. Given the high levels of diazinon removal, the SMC formed by four Streptomyces strains has the potential to be used to treat the diazinon present in environmental matrices.

Probing non polar interstellar molecules through their protonated form: Detection of protonated cyanogen (NCCNH+).

Cyanogen (NCCN) is the simplest member of the series of dicyanopolyynes. It has been hypothesized that this family of molecules can be important constituents of interstellar and circumstellar media, although the lack of a permanent electric dipole moment prevents its detection through radioastronomical techniques. Here we present the first solid evidence of the presence of cyanogen in interstellar clouds through the detection of its protonated form toward the cold dark clouds TMC-1 and L483. Protonated cyanogen (NCCNH+) has been identified through the J = 5 - 4 and J = 10 - 9 rotational transitions using the 40m radiotelescope of Yebes and the IRAM 30m telescope. We derive beam averaged column densities for NCCNH+ of (8.6 ± 4.4) × 1010 cm-2 in TMC-1 and (3.9 ± 1.8) × 1010 cm-2 in L483, which translate to fairly low fractional abundances relative to H2, in the range (1-10) × 10-12. The chemistry of protonated molecules in dark clouds is discussed, and it is found that, in general terms, the abundance ratio between the protonated and non protonated forms of a molecule increases with increasing proton affinity. Our chemical model predicts an abundance ratio NCCNH+/NCCN of ~ 10-4, which implies that the abundance of cyanogen in dark clouds could be as high as (1-10) × 10-8 relative to H2, i.e., comparable to that of other abundant nitriles such as HCN, HNC, and HC3N.

Evaluation of biodegradable polymers as encapsulating agents for the development of a urea controlled-release fertilizer using biochar as support material.

Biochar constitutes a promising support material for the formulation of controlled-release fertilizers (CRFs). In this study we evaluated the effect of different polymeric materials as encapsulating agents to control nitrogen (N) leaching from biochar based CRFs. Nitrogen impregnation onto biochar was performed in a batch reactor using urea as N source. The resulting product was encapsulated by using sodium alginate (SA), cellulose acetate (CA) and ethyl cellulose (EC). Leaching potential was studied in planted and unplanted soil columns, monitoring nitrate, nitrite, ammonium and urea concentrations. After 90 days, plants were removed from the soil columns and plant yield was evaluated. It was observed that the ammonium concentration in leachates presented a maximum concentration for all treatments at day 22. The highest concentration of N in the leachates was the nitrate form. The crop yield was negatively affected by all developed CRFs using biochar compared with the traditional fertilization.

Are white-rot fungi a real biotechnological option for the improvement of environmental health?

The use of white-rot fungi as a biotechnological tool for cleaning the environment of recalcitrant pollutants has been under evaluation for several years. However, it is still not possible to find sufficiently detailed investigations of this subject to conclude that these fungi can decontaminate the environment. In the present review, we have summarized and discussed evidence about the potential of white-rot fungi to degrade such pollutants as polycyclic aromatic hydrocarbons, dyes or antibiotics as an example of the complex structures that these microorganisms can attack. This review also discusses field experiment results and limitations of white-rot fungi trials from contaminated sites. Moreover, the use of catabolic potential of white-rot fungi in biopurification systems (biobeds) is also discussed. The current status and future perspectives of white-rot fungi, as a viable biotechnological alternative for improvement of environmental health are noted.

Enzymes as useful tools for environmental purposes.

In the environment enzymes may play important and different roles at least in three cases: as main agents (as isolated, cell-bound or immobilized enzymes) in charge of either the transformation and/or degradation of compounds polluting the environment and the restoration of the polluted environment; as reliable and sensitive tools to detect and measure the amount and concentration of pollutants before, during and after the restoration process; as reliable, easy and sensitive indicators of quality and health status of the environment subjected to the restoration process. To our knowledge papers or reviews integrating findings on these three functions of enzymes are missing in literature. Therefore the main scope of the present paper is to briefly encompass general and specific concepts about roles of enzymes as decontaminating agents, pollutant assaying agents and indicators of environment safety. Examples chosen among those published very recently, supporting and confirming peculiarities, features, and performance of enzymatic agents will be illustrated.

Simulators help improve student confidence to acquire skills in urology.

OBJECTIVE: To know the level of confidence of fifth year medical students in order to perform maneuvers in bladder catheterization and rectal examination before and after training with simulators. To be able to assess student satisfaction regarding the use of the simulation as a learning method. MATERIAL AND METHODS: The study was conducted in the Simulation Center of the Faculty of Medicine. A total of 173 students who completed a practical workshop on the subject of Urology participated. The students were asked to answer anonymous questionnaires on their level of confidence in performing a bladder catheterization and rectal examination before and after the workshop as well as their satisfaction in using the simulation as a training tool. The workshops were organized using groups of 10 students. A teacher or a resident in that area of expertise supervised each student individually, resolving their doubts and teaching them the proper technique. RESULTS: All the evaluations made on the different abilities were significantly higher after training (P<.001). Significant differences were found in the confidence level between men and women before the training regarding male urethral catheterization maneuvers and recognition of normal or pathological prostate, The confidence level was lower in women (P<.05). These differences disappeared after training. The level of overall satisfaction with the workshop was high, going from 4.47 ± 0.9 to a maximum score of 5. CONCLUSIONS: Simulation is a training method that helps improve the confidence of the medical student in performing a bladder catheterization and digital rectal examination.

[Confidence assessment among medical students trained with an obstetric simulator]. / Confianza de los estudiantes de medicina en el aprendizaje de la exploración obstétrica con simuladores.

BACKGROUND: To determine the degree of confidence of students regarding obstetric examination before and after training with simulators, and assess their satisfaction with the use of simulation as a tool for acquiring skills. METHODS: Training groups involved 10 students per lecturer. Students learned how to perform Leopold's manoeuvres and measure uterine height with a simulator. The instructor supervised each physical exam. Surveys by those students who had previously done an internship at the Department of Obstetric & Gynecology were not taken into account. RESULTS: Students' confidence in performing the procedures improved significantly (p≤0.001) after the intervention.

Atrazine dissipation and its impact on the microbial communities and community level physiological profiles in a microcosm simulating the biomixture of on-farm biopurification system.

The effects of repeated atrazine application (40 mg a.i.kg(-1)) on its degradation, microbial communities and enzyme activities were studied in a peat based biomixture composed by straw, soil and peat in the volumetric proportions of 2:1:1 that can be used in on-farm biopurification system. Atrazine removal efficiency was high (96%, 78% and 96%) after each atrazine application and did not show a lag phase. Microbial enzyme activities were reduced significantly with atrazine application but rapidly recovered. Microbial diversity obtained by BiologEcoplate was similar after the first and second atrazine application. However, an inhibitory effect was observed after the third application. After each atrazine application, culturable fungi were reduced, but rapidly recovered without significant changes in culturable bacteria and actinomycetes compared to the control. Denaturing gradient gel electrophoresis (DGGE) patterns demonstrated that microbial community structure remained relatively stable in time when compared to the controls. In conclusion, our results demonstrated that after successive ATZ applications, the peat based biomixture had a good degradation capacity. Moreover, microbiological assays demonstrated the robustness of the peat based biomixture from a microbiological point of view to support pesticide degradation.

Degradation of pesticide mixture on modified matrix of a biopurification system with alternatives lignocellulosic wastes.

The biobed systems were designed to retain and to degrade pesticides through the properties of a biomixture composed of straw (ST), topsoil and peat (PT) 2:1:1 v/v. The ST is the main substrate in the biomixture, as it allows the proliferation of fungi that promotes pesticide degradation. The use of readily available components in the biomixture is an important aspect to build a biobed. Therefore, potential use of readily available wastes as barley husk (BH), sawdust (SW) and oat husk (OH), as total or partial substitutes of ST were tested in pesticide degradation studies. Metabolite formation and the biological activities were also evaluated. Biomixture composed of OH was highly efficient in pesticide degradation, with t½ values of 28.6, 58.9 and 26.8 d for atrazine (ATZ), chlorpyrifos (CHL) and isoproturon (ISP). On the other hand, comparable for degrading capacities with the ST based biomixture were obtained with SW and BH, but only as partial replacement. Contrarily, high t½ values (more than 100 d) were obtained in biomixtures with total substitution of ST by SW or BH. Metabolite formation was observed in all biomixtures tested, but without clear formation patterns. Moreover, high and stable biological activity was observed in the biomixtures composed of OH. Therefore, our results demonstrated that ST can be partial or totally replaced by OH in the biomixture allowing an efficient degradation of pesticide mixture. However, it is recommended that ST can be only partially replaced by BH and SW in the biomixture to allow efficient pesticide degradation.