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Abstract Aim We tested the effects of a cancer cachexia-anorexia sydrome upon the balance of anti and pro-inflammatory cytokines in the hypothalamus of sedentary or trained tumour-bearing (Walker-256 carcinosarcoma) rats. Methods Animals were randomly assigned to a sedentary control (SC), sedentary tumour-bearing (ST), and sedentary pair-fed (SPF) groups or, exercised control (EC), exercised tumour-bearing (ET) and exercised pair-fed (EPF) groups. Trained rats ran on a treadmill (60%VO2max) for 60 min/d, 5 days/wk, for 8 wks. We evaluated food intake, leptin and cytokine (TNF-α, IL1β) levels in the hypothalamus. Results The cumulative food intake and serum leptin concentration were reduced in ST compared to SC. Leptin gene expression in the retroperitoneal adipose tissue (RPAT) was increased in SPF in comparison with SC and ST, and in the mesenteric adipose tissue (MEAT) the same parameter was decreased in ST in relation to SC. Leptin levels in RPAT and MEAT were decreased in ST, when compared with SC. Exercise training was also able to reduce tumour weight when compared to ST group. In the hypothalamus, IL-1β and IL-10 gene expression was higher in ST than in SC and SPF. Cytokine concentration in hypothalamus was higher in ST (TNF-α and IL-1β, p < 0.05), compared with SC and SPF. These pro-inflammatory cytokines concentrations were restored to control values (p < 0.05), when the animals were submitted to endurance training. Conclusion Cancer-induced anorexia leads towards a pro-inflammatory state in the hypothalamus, which is prevented by endurance training which induces an anti-inflammatory state, with concomitant decrease of tumour weight.

Abstract Background The three trypanosomatids pathogenic to men, Trypanosoma cruzi, Trypanosoma brucei and Leishmania major, are etiological agents of Chagas disease, African sleeping sickness and cutaneous leishmaniasis, respectively. The complete sequencing of these trypanosomatid genomes represented a breakthrough in the understanding of these organisms. Genome sequencing is a step towards solving the parasite biology puzzle, as there are a high percentage of genes encoding proteins without functional annotation. Also, technical limitations in protein expression in heterologous systems reinforce the evident need for the development of a high-throughput reverse genetics platform. Ideally, such platform would lead to efficient cloning and compatibility with various approaches. Thus, we aimed to construct a highly efficient cloning platform compatible with plasmid vectors that are suitable for various approaches. Results We constructed a platform with a flexible structure allowing the exchange of various elements, such as promoters, fusion tags, intergenic regions or resistance markers. This platform is based on Gateway® technology, to ensure a fast and efficient cloning system. We obtained plasmid vectors carrying genes for fluorescent proteins (green, cyan or yellow), and sequences for the c-myc epitope, and tandem affinity purification or polyhistidine tags. The vectors were verified by successful subcellular localization of two previously characterized proteins (TcRab7 and PAR 2) and a putative centrin. For the tandem affinity purification tag, the purification of two protein complexes (ribosome and proteasome) was performed. Conclusions We constructed plasmids with an efficient cloning system and suitable for use across various applications, such as protein localization and co-localization, protein partner identification and protein expression. This platform also allows vector customization, as the vectors were constructed to enable easy exchange of its elements. The development of this high-throughput platform is a step closer towards large-scale trypanosome applications and initiatives.

Juan S Izquierdo-Condoy,1 Miguel Angel Montiel-Alfonso,2 Humberto Alejandro Nati-Castillo,3 Ruth Saucedo,4 Damary S Jaramillo-Aguilar,5 Camila Nanjari-Barrientos,6 Carolina García-Arévalo,7 Daniel Rivera-Flores,8 Maria Ibanesi Díaz Batista,9 Valentina Loaiza-Guevara,10 Esteban Ortiz-Prado1 1OneHealth Global Research Group, Universidad de las Américas, Quito, Ecuador; 2Faculty of Health Sciences, Universidad Católica Nuestra Señora de la Asunción, Asunción, Paraguay; 3Faculty of Health Sciences, Universidad del Quindío, Armenia, Colombia; 4Scientific Committee, Bolivian Association of Scientific Societies of Medical Students (ABOLSCEM), Santa Cruz, Bolivia; 5Faculty of Medical Sciences, University of Cuenca, Cuenca, Ecuador; 6Faculty of Medicine, University of Valparaiso, Valparaiso, Chile; 7Faculty of Medicine, University of Panama, Panama City, Panama; 8Faculty of Medical Sciences, Universidad Nacional Autónoma de Honduras, Tegucigalpa, Honduras; 9Faculty of Health Sciences, Universidad Autonoma de Santo Domingo, Santo Domingo, República Dominicana; 10Faculty of Medicine, Institución Universitaria Visión de las Américas, Pereira, ColombiaCorrespondence: Esteban Ortiz-Prado, OneHealth Global Research Group, Universidad de las Américas, Calle de los Colimes y Avenida De los Granados, Quito, 170137, Ecuador, Tel +593995760693, Email e.ortizprado@gmail.comPurpose: The objective of this study was to describe the level of knowledge, perceptions, and practices in relation to risks and disasters in medical schools in Latin America and the Caribbean.Participants and Methods: Multicenter, observational, analytical, non-probabilistic convenience sample study with 2546 medical students in 9 countries of Latin America and the Caribbean. An online survey was conducted between October 2020 and November 2020, using an instrument validated in each country to assess knowledge, perceptions, and practices regarding risk and disaster prevention measures. Frequencies, percentages, mean and standard deviation (SD) were used for descriptive analysis. Differences resulting from the relationship between the variables studied and the level of knowledge were obtained using the Chi-square test. P-value < 0.05 was accepted as statistically significant for all analyses.Results: The highest proportion of responses came from women, third-semester students, and those studying in public universities. Students from Colombia and Honduras had the highest percentage of high levels of knowledge about disasters, while Peruvian students had the highest percentage of low levels of knowledge. Women and students from public universities showed a higher proportion of high levels of knowledge. 52.7% considered that they live in a country with a medium risk of natural disasters, while 91.2% said that Latin American and Caribbean countries are not prepared to face natural disasters. Only 43.6% believe they are prepared to help in the event of a natural disaster.Conclusion: Most of medical students from Latin America and Latin America and the Caribbean have high and medium level of knowledge in risks and disasters. However, the implementation of disaster training programs for medical students has the potential to improve the preparedness, knowledge, and skills that are important for medical personnel to improve their self-confidence, and their ability to respond, resulting in more effective systems.Keywords: disaster medicine, knowledge, perceptions, practices, students, medical, Latin America, Caribbean

The increasing incidence of neurodegenerative and other diseases is considered to involve an excessive production of reactive oxygen species (ROS). Water supplies are often characterized by excessive organic waste that is decomposed by bacteria, using dissolved oxygen, leading to oxygen depletion. The potassium content of these waters may also affect negatively the mitochondrial metabolism and cellular ROS formation. This work focused on characterizing mitochondrial autofluorescence changes, with flavoprotein origin, and fluorescence ROS signals measured using the 2',7'-dichlorodihydrofluorescein diacetate indicator H2DCFDA. All signals were evoked by hypoxia or by the depolarizing agent KCl (20 mM), at the hippocampal mossy fiber synapses of CA3 area. It was observed that both hypoxia and KCl-induced depolarization elicited a small rise in the autofluorescence and ROS changes. The hypoxia-induced signals were maintained upon normal reoxygenation, but of those evoked by KCl, the autofluorescence signals recovered during washout, while the ROS changes were irreversible.

Objective: Families affected by asthma report difficulty adhering to care regimens because of high medication costs, coupled with increased cost sharing required by some insurance plans. To inform efforts to support adherence, we conducted a qualitative study to explore how families manage asthma care costs., Methods: We conducted phone interviews with commercially-insured, US adults ( n  = 59) who had asthma and/or a child with asthma. Our purposive sample included participants with high- and low/no-deductible health plans. We analyzed data using thematic content analysis to identify strategies for managing asthma care costs and to assess strategies' implications for adherence., Results: Our analysis identified four overarching strategies for managing asthma care costs. First, participants used prevention strategies to avoid costly acute care by minimizing exposure to asthma triggers and adhering strictly to preventive medication regimens. Second, participants used shopping strategies to reduce costs, including by comparing medication prices across pharmacies, using medication coupons or free samples, and switching to lower-cost medications. Third, budgeting strategies involved putting aside funds, including in tax-exempt health savings accounts, or taking on debt to pay for care. Finally, some participants sought to reduce costs by forgoing recommended care, including by skipping medication doses or replacing prescribed medications with alternative therapies., Conclusion: Commercially-insured families use a wide range of strategies to manage asthma care costs, with both positive and negative implications for adherence. Our typology of asthma cost management strategies can inform insurance redesign and other interventions to help families safely reduce costs and maximize adherence to recommended care.

Competing Interests: None

Short-term variability in condition factor: relative condition factor Kn; biochemical condition: RNA:DNA and protein content; and instantaneous growth rates were determined in estuarine and coastal fish. Dicentrarchus labrax, Solea senegalensis and Pomatoschistus microps were sampled in the Tejo estuary, while Trachurus trachurus was sampled in an adjacent shallow coastal area. Variation of condition indices was more frequent at the week scale (sampling periods with fortnight intervals) than at the daily scale (consecutive days in each sampling period) in all species. Water temperature was correlated with biochemical indices, while salinity showed no effect, evidencing the influence of environmental short-term variation (temperature) on biochemical condition in natural populations. Yet, decreasing individual variability in fish condition was observed along the sampled weeks, resulting in a more homogeneous condition of populations, particularly for T. trachurus likely due to a more stable coastal environment. Biochemical indices proved to be sensitive to short-term environmental variability, despite species-specific responses., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Estuaries are threatened by intense and continuously increasing human activities. Here we estimated the sensitivity of fish assemblages in a set of estuaries distributed worldwide (based on species vulnerability and resilience), and the exposure to cumulative stressors and coverage by protected areas in and around those estuaries (from marine, estuarine and freshwater ecosystems, due to their connectivity). Vulnerability and resilience of estuarine fish assemblages were not evenly distributed globally and were driven by environmental features. Exposure to pressures and extent of protection were also not evenly distributed worldwide. Assemblages with more vulnerable and less resilient species were associated with estuaries in higher latitudes (in particular Europe), and with higher connectivity with the marine ecosystem, moreover such estuaries were generally under high intensity of pressures but with no concomitant increase in protection. Current conservation schemes pay little attention to species traits, despite their role in maintaining ecosystem functioning and stability. Results emphasize that conservation is weakly related with the global distribution of sensitive fish species in sampled estuaries, and this shortcoming is aggravated by their association with highly pressured locations, which appeals for changes in the global conservation strategy (namely towards estuaries in temperate regions and highly connected with marine ecosystems).

Marine Protected Areas (MPAs) have been widely proposed as a fisheries management tool in addition to their conservation purposes. Despite this, few studies have satisfactorily assessed the dynamics of fishers' adaptations to the loss of fishing grounds. Here we used data from before, during and after the implementation of the management plan of a temperate Atlantic multiple-use MPA to examine the factors affecting the spatial and temporal distribution of different gears used by the artisanal fishing fleet. The position of vessels and gear types were obtained by visual surveys and related to spatial features of the marine park. A hotspot analysis was conducted to identify heavily utilized patches for each fishing gear and time period. The contribution of individual vessels to each significant cluster was assessed to better understand fishers' choices. Different fisheries responded differently to the implementation of protection measures, with preferred habitats of target species driving much of the fishers' choices. Within each fishery, individual fishers showed distinct strategies with some operating in a broader area whereas others kept preferred territories. Our findings are based on reliable methods that can easily be applied in coastal multipurpose MPAs to monitor and assess fisheries and fishers responses to different management rules and protection levels. This paper is the first in-depth empirical study where fishers' choices from artisanal fisheries were analysed before, during and after the implementation of a MPA, thereby allowing a clearer understanding of the dynamics of local fisheries and providing significant lessons for marine conservation and management of coastal systems.

The extent to which host biology, ecology and phylogeny determine the diversity of macroparasite assemblages has been investigated in recent years in several taxa, including fish. However, consensus has not been reached probably as a result of data being collected from different sources, different temporal scales or host and parasite biogeography and phylogeny having greater influence than expected. The present study evaluates the relative importance of 27 biological, ecological and phylogenetic characteristics of 14 flatfish species on the diversity of their ecto- and endoparasite assemblages, comprising a total of 53 taxa. Redundancy analyses were applied to the mean abundance of each parasite taxa infecting each host and to the richness, taxonomic distinctness and variance in taxonomic distinctness calculated for each assemblage within each host. Only a few host characteristics contributed significantly to the observed patterns: host distribution was more important in determining the type and mean abundance of ectoparasites present in an assemblage, whereas diversity of these assemblages were mainly related to the host's maximum size. Endoparasite mean abundance and diversity were mostly influenced by the number of food items ingested and by the presence of Crustacea and Polychaeta in the diet. However, the sympatric occurrence of related hosts also played an important role in the diversity values found in macroparasite assemblages. Results showed that a host characteristic has different importance according to the host-parasite relationship being examined, suggesting an important role for host-parasite co-evolution on the diversity of extant macroparasite assemblages.