Advanced Imaging Methodology in Bacterial Biofilms with a Fluorescent Enzymatic Sensor for pepN Activity.

This research explores the use of the pepN activity fluorescent sensor DCM-Ala in bacterial biofilms, emphasizing its significance due to the critical role of biofilms in various biological processes. Advanced imaging techniques were employed to visualize pepN activity, introducing a novel approach to examining biofilm maturity. We found that the overexpression of pepN increases the ability of E. coli to form biofilm. The findings demonstrate varying levels of pepN activity throughout biofilm development, suggesting potential applications in biofilm research and management. The results indicate that the fluorescent emission from this sensor could serve as a reliable indicator of biofilm maturity, and the imaging techniques developed could enhance our understanding and control of biofilm-related processes. This work highlights the importance of innovative methods in biofilm study and opens new avenues for utilizing chemical emissions in biofilm management.

IMP3 Immunohistochemical Expression Is Related with Progression and Metastases in Xenografted and Cutaneous Melanomas.

INTRODUCTION: Insulin-like growth factor-II messenger RNA-binding protein-3 (IMP3) over-expression is a predictor of tumor recurrence and metastases in some types of human melanoma. Our objective was to evaluate the immunohistochemical expression of IMP3 and other molecules related to tumor prognosis in melanoma-xeno-tumors undergoing treatment. We test the effect of radiotherapy (RT) and mesenchymal stromal cells (MSCs) treatment, analyzing the tumorigenic and metastatsizing capacity in a mice melanoma xenograft model. MATERIALS AND METHODS: We inoculated A375 and G361 human melanoma cell lines into NOD/SCID gamma mice (n = 64). We established a control group, a group treated with MSCs, a group treated with MSCs plus RT, and a group treated with RT. We assessed the immunohistochemical expression of IMP3, E-cadherin, N-cadherin, PARP1, HIF-1α, and the proliferation marker Ki-67. Additionally, we performed a retrospective study including 114 histological samples of patients diagnosed with malignant cutaneous superficial spreading melanoma (n = 104) and nodular melanoma (n = 10) with at least 5 years of follow-up. RESULTS: Most morphological and immunohistochemical features show statistically significant differences between the 2 cell lines. The A375 cell line induced the formation of metastases, while the G361 cell line provoked tumor formation but not metastases. All three treatments reduced the cell proliferation evaluated by the Ki-67 nuclear antigen (p = 0.000, one-way ANOVA test) and reduced the number of metastases (p = 0.004, one-way ANOVA test). In addition, the tumor volumes reduced in comparison with the control groups, 31.74% for RT + MSCs in the A357 tumor cell line, and 89.84% RT + MSCs in the G361 tumor cell line. We also found that IMP3 expression is associated with greater tumor aggressiveness and was significantly correlated with cell proliferation (measured by the expression of Ki-67), the number of metastases, and reduced expression of adhesion molecules. CONCLUSIONS: The combined treatment of RT and MSCs on xenografted melanomas reduces tumor size, metastases frequency, and the epithelial to mesenchymal transition/PARP1 metastatic phenotype. This treatment also reduces the expression of molecules related to cellular proliferation (Ki-67), molecules that facilitate the metastatic process (E-cadherin), and molecules related with prognosis (IMP3).

Ratiometric Two-Photon Near-Infrared Probe to Detect DPP IV in Human Plasma, Living Cells, Human Tissues, and Whole Organisms Using Zebrafish.

DPP IV, otherwise known as CD26 lymphocyte T surface antigen, is a transmembrane glycoprotein also found in circulation in the blood. It plays an important role in several processes like glucose metabolism and T-cell stimulation. Moreover, it is overexpressed in renal, colon, prostate, and thyroid human carcinoma tissues. It can also serve as a diagnostic in patients with lysosomal storage diseases. The biological and clinical importance of having readouts for the activity of this enzyme, in physiological and disease conditions, has led us to design a near-infrared (NIR) fluorimetric probe that also has the characteristics of being ratiometric and excitable by two simultaneous NIR photons. The probe consists of assembling an enzyme recognition group (Gly-Pro) (Mentlein, 1999; Klemann et al., 2016) on the two-photon (TP) fluorophore (derivative of dicyanomethylene-4H-pyran, DCM-NH2) disturbing its NIR characteristic internal charge transfer (ICT) emission spectrum. When the dipeptide group is released by the DPP IV-specific enzymatic action, the donor-acceptor DCM-NH2 is restored, forming a system that shows high ratiometric fluorescence output. With this new probe, we have been able to detect, quickly and efficiently, the enzymatic activity of DPP IV in living cells, human tissues, and whole organisms, using zebrafish. In addition, due to the possibility of being excited by two photons, we can avoid the autofluorescence and subsequent photobleaching that the raw plasma has when it is excited by visible light, achieving detection of the activity of DPP IV in that medium without interference.

Lifting COVID-19 mitigation measures in Spain (May-June 2020).

INTRODUCTION: The state of alarm was declared in Spain due to the COVID-19 epidemic on March 14, 2020, and established population confinement measures. The objective is to describe the process of lifting these mitigation measures. METHODS: The Plan for the Transition to a New Normality, approved on April 28, contained four sequential phases with progressive increase in socio-economic activities and population mobility. In parallel, a new strategy for early diagnosis, surveillance and control was implemented. A bilateral decision mechanism was established between the Spanish Government and the autonomous communities (AC), guided by a set of qualitative and quantitative indicators capturing the epidemiological situation and core capacities. The territorial units were established ad-hoc and could be from Basic Health Zones to entire AC. RESULTS: The process run from May 4 to June 21, 2020. AC implemented plans for reinforcement of core capacities. Incidence decreased from a median (50% of territories) of 7.4 per 100,000 in 7 days at the beginning to 2.5 at the end. Median PCR testing increased from 53% to 89% of suspected cases and PCR total capacity from 4.5 to 9.8 per 1000 inhabitants weekly; positivity rate decreased from 3.5% to 1.8%. Median proportion of cases with traced contacts increased from 82% to 100%. CONCLUSION: Systematic data collection, analysis, and interterritorial dialogue allowed adequate process control. The epidemiological situation improved but, mostly, the process entailed a great reinforcement of core response capacities nation-wide, under common criteria. Maintaining and further reinforcing capacities remained crucial for responding to future waves.

Lifting COVID-19 mitigation measures in Spain (May-June 2020).

Introduction: The state of alarm was declared in Spain due to the COVID-19 epidemic on March 14, 2020, and established population confinement measures. The objective is to describe the process of lifting these mitigation measures. Methods: The Plan for the Transition to a New Normality, approved on April 28, contained four sequential phases with progressive increase in socio-economic activities and population mobility. In parallel, a new strategy for early diagnosis, surveillance and control was implemented. A bilateral decision mechanism was established between the Spanish Government and the autonomous communities (AC), guided by a set of qualitative and quantitative indicators capturing the epidemiological situation and core capacities. The territorial units were established ad-hoc and could be from Basic Health Zones to entire AC. Results: The process run from May 4 to June 21, 2020. AC implemented plans for reinforcement of core capacities. Incidence decreased from a median (50% of territories) of 7.4 per 100,000 in 7 days at the beginning to 2.5 at the end. Median PCR testing increased from 53% to 89% of suspected cases and PCR total capacity from 4.5 to 9.8 per 1000 inhabitants weekly; positivity rate decreased from 3.5% to 1.8%. Median proportion of cases with traced contacts increased from 82% to 100%. Conclusion: Systematic data collection, analysis, and interterritorial dialogue allowed adequate process control. The epidemiological situation improved but, mostly, the process entailed a great reinforcement of core response capacities nation-wide, under common criteria. Maintaining and further reinforcing capacities remained crucial for responding to future waves.

Introducción: El 14 de marzo de 2020 España declaró el estado de alarma por la pandemia por COVID-19 incluyendo medidas de confinamiento. El objetivo es describir el proceso de desescalada de estas medidas. Métodos: Un plan de transición hacia una nueva normalidad, del 28 de abril, incluía 4 fases secuenciales incrementando progresivamente las actividades socioeconómicas y la movilidad. Concomitantemente, se implementó una nueva estrategia de diagnóstico precoz, vigilancia y control. Se estableció un mecanismo de decisión bilateral entre Gobierno central y comunidades autónomas (CCAA), guiado por un panel de indicadores cualitativos y cuantitativos de la situación epidemiológica y las capacidades básicas. Las unidades territoriales evaluadas comprendían desde zonas básicas de salud hasta CCAA. Resultados: El proceso se extendió del 4 de mayo al 21 de junio y se asoció a planes de refuerzo de las capacidades en las CCAA. La incidencia disminuyó de una mediana inicial de 7,4 por 100.000 en 7 días a 2,5 al final del proceso. La mediana de pruebas PCR aumentó del 53% al 89% de los casos sospechosos, y la capacidad total de 4,5 a 9,8 pruebas semanales por 1.000 habitantes; la positividad disminuyó del 3,5% al 1,8%. La mediana de casos con contactos trazados aumentó del 82% al 100%. Conclusión: La recogida y análisis sistemático de información y el diálogo interterritorial logaron un adecuado control del proceso. La situación epidemiológica mejoró, pero sobre todo, se aumentaron las capacidades, en todo el país y con criterios comunes, cuyo mantenimiento y refuerzo fue clave en olas sucesivas.

New bioisosteric sulphur-containing choline kinase inhibitors with a tracked mode of action.

Since the identification of human choline kinase as a protein target against cancer progression, many compounds have been designed to inhibit its function and reduce the biosynthesis of phosphatidylcholine. Herein, we propose a series of bioisosteric inhibitors that are based on the introduction of sulphur and feature improved activity and lipophilic/hydrophilic balance. The evaluation of the inhibitory and of the antiproliferative properties of the PL (dithioethane) and FP (disulphide) libraries led to the identification of PL 48, PL 55 and PL 69 as the most active compounds of the series. Docking analysis using FLAP suggests that for hits to leads, binding mostly involves an interaction with the Mg2+ cofactor, or its destabilization. The most active compounds of the two series are capable of inducing apoptosis following the mitochondrial pathway and to significantly reduce the expression of anti-apoptotic proteins such as the Mcl-1. The fluorescence properties of the compounds of the PL library allowed the tracking of their mode of action, while PAINS (Pan Assays Interference Structures) filtration databases suggest the lack of any unspecific biological response.

Hypothalamic orexinergic neuron changes during the hibernation of the Syrian hamster.

Hibernation in small mammals is a highly regulated process with periods of torpor involving drops in body temperature and metabolic rate, as well as a general decrease in neural activity, all of which proceed alongside complex brain adaptive changes that appear to protect the brain from extreme hypoxia and low temperatures. All these changes are rapidly reversed, with no apparent brain damage occurring, during the short periods of arousal, interspersed during torpor-characterized by transitory and partial rewarming and activity, including sleep activation, and feeding in some species. The orexins are neuropeptides synthesized in hypothalamic neurons that project to multiple brain regions and are known to participate in the regulation of a variety of processes including feeding behavior, the sleep-wake cycle, and autonomic functions such as brown adipose tissue thermogenesis. Using multiple immunohistochemical techniques and quantitative analysis, we have characterized the orexinergic system in the brain of the Syrian hamster-a facultative hibernator. Our results revealed that orexinergic neurons in this species consisted of a neuronal population restricted to the lateral hypothalamic area, whereas orexinergic fibers distribute throughout the rostrocaudal extent of the brain, particularly innervating catecholaminergic and serotonergic neuronal populations. We characterized the changes of orexinergic cells in the different phases of hibernation based on the intensity of immunostaining for the neuronal activity marker C-Fos and orexin A (OXA). During torpor, we found an increase in C-Fos immunostaining intensity in orexinergic neurons, accompanied by a decrease in OXA immunostaining. These changes were accompanied by a volume reduction and a fragmentation of the Golgi apparatus (GA) as well as a decrease in the colocalization of OXA and the GA marker GM-130. Importantly, during arousal, C-Fos and OXA expression in orexinergic neurons was highest and the structural appearance and the volume of the GA along with the colocalization of OXA/GM-130 reverted to euthermic levels. We discuss the involvement of orexinergic cells in the regulation of mammalian hibernation and, in particular, the possibility that the high activation of orexinergic cells during the arousal stage guides the rewarming as well as the feeding and sleep behaviors characteristic of this phase.

Thermal and Rheological Properties of Hydrophobic Nanosilica in Sunflower Oil Suspensions at High Pressures.

Nowadays, the reduction of the environmental impact associated with the operation of the oil industry is a primary concern. A growing trend is to develop low-toxicity formulations based on biodegradable components. In this sense, vegetable oils structured with nanomaterials could be an alternative to mineral or synthetic oils for sustainable fluid formulations. Hydrophobic fumed silica nanoparticles have the capability to change the rheological behavior of oil in suspensions, providing a large variety of non-Newtonian behaviors over a wide range of temperatures, from shear-thinning to gel-like, depending on the concentration and the nanosilica's hydrophobicity, that permits the design of fluids with selected characteristic and applications. This work explores the microstructure and the rheological behavior of hydrophobic fumed silica dispersed in a sunflower oil as a function of temperature and pressure. The results suggest that the suspensions of hydrophobic silica in sunflower oil reveals appropriate rheological and thermal properties over a wide range of temperatures and pressures to serve as components of sustainable drilling fluids.

Lifting COVID-19 mitigation measures in Spain (May-June 2020).

INTRODUCTION: The state of alarm was declared in Spain due to the COVID-19 epidemic on March 14, 2020, and established population confinement measures. The objective is to describe the process of lifting these mitigation measures. METHODS: The Plan for the Transition to a New Normality, approved on April 28, contained four sequential phases with progressive increase in socio-economic activities and population mobility. In parallel, a new strategy for early diagnosis, surveillance and control was implemented. A bilateral decision mechanism was established between the Spanish Government and the autonomous communities (AC), guided by a set of qualitative and quantitative indicators capturing the epidemiological situation and core capacities. The territorial units were established ad-hoc and could be from Basic Health Zones to entire AC. RESULTS: The process run from May 4 to June 21, 2020. AC implemented plans for reinforcement of core capacities. Incidence decreased from a median (50% of territories) of 7.4 per 100,000 in 7 days at the beginning to 2.5 at the end. Median PCR testing increased from 53% to 89% of suspected cases and PCR total capacity from 4.5 to 9.8 per 1000 inhabitants weekly; positivity rate decreased from 3.5% to 1.8%. Median proportion of cases with traced contacts increased from 82% to 100%. CONCLUSION: Systematic data collection, analysis, and interterritorial dialogue allowed adequate process control. The epidemiological situation improved but, mostly, the process entailed a great reinforcement of core response capacities nation-wide, under common criteria. Maintaining and further reinforcing capacities remained crucial for responding to future waves.

New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses.

A new chloride-sensitive red fluorescent protein derived from Entacmaea quadricolor is described. We found that mBeRFP exhibited moderate sensitivity to chloride and, via site-directed mutagenesis (S94V and R205Y), we increased the chloride affinity by more than an order of magnitude (kd = 106 ± 6 mM) at physiological pH. In addition, cis-trans isomerization of the chromophore produces a dual emission band with different chloride sensitivities, which allowed us to develop a ratiometric methodology to measure intracellular chloride concentrations.

[Municipalism and community health: transforming through local government. SESPAS Report 2018]. / Municipalismo y salud comunitaria: transformar desde los ayuntamientos. Informe SESPAS 2018.

Municipalities and local government are an essential element to promote public policies that improve people's health, and impact the social determinants of health, through developing approaches which incorporate equity, community engagement and intersectoral partnership at their core. The objective of this article is to analyze the barriers and opportunities that can be encountered within local governments when interventions aimed at improving people's health are developed and implemented. The evolution of the city councils, their competence frameworks and the current socio-political context are described, and three key tools for local action are proposed: intersectorality, community engagement and working in partnership with other administrations. Global strategies such as the implementation of the "community health" and "health in all policies" approach through cross-sectoral networks/partnerships are described, emphasizing the importance of developing formal and informal structures or processes of community engagement and designing local health action plans in cooperation with other administrations, such as regional and national governing bodies. To continue supporting these forms of local governance, we propose adapting the local administrations to the new social realities, with more horizontal and flexible organization models and resource allocation, integrating efficiency and evaluation processes, reclaiming local autonomy, and building local governance through networks and alliances.

Municipalismo y salud comunitaria: transformar desde los ayuntamientos. Informe SESPAS 2018 / Municipalism and community health: transforming through local government. SESPAS Report 2018

El ámbito municipal, y con él el municipalismo, se configura como un elemento fundamental para el impulso de políticas públicas que mejoren la salud de las poblaciones, incidan sobre los determinantes sociales de la salud y tengan en cuenta de forma transversal la equidad, la participación ciudadana y la intersectorialidad. El objetivo de este artículo es analizar las dificultades y las oportunidades que ofrece el ámbito municipal para poner en marcha iniciativas locales e incidir en la salud de las poblaciones. En él se describe la evolución de los ayuntamientos y de sus marcos competenciales en el contexto sociopolítico actual. Se proponen tres herramientas clave para la acción local: la intersectorialidad, la participación comunitaria, y las redes y el trabajo conjunto con otras administraciones. Se ahonda en la puesta en marcha de estrategias globales como la implementación del enfoque de «salud comunitaria» y «salud en todas políticas» mediante mesas intersectoriales, la formación de estructuras o procesos formales e informales de participación comunitaria, y la realización de planes de salud local en cooperación con otras administraciones. Como medidas para seguir avanzando se propone adaptar las administraciones locales a las nuevas realidades sociales, crear modelos de organización más horizontales y flexibles, recuperar la autonomía local, dotar a los ayuntamientos de recursos, incorporar la evaluación y la búsqueda de la eficiencia, y construir redes y alianzas de gobernanza local

Municipalities and local government are an essential element to promote public policies that improve people's health, and impact the social determinants of health, through developing approaches which incorporate equity, community engagement and intersectoral partnership at their core. The objective of this article is to analyze the barriers and opportunities that can be encountered within local governments when interventions aimed at improving people's health are developed and implemented. The evolution of the city councils, their competence frameworks and the current socio-political context are described, and three key tools for local action are proposed: intersectorality, community engagement and working in partnership with other administrations. Global strategies such as the implementation of the "community health" and "health in all policies" approach through cross-sectoral networks/partnerships are described, emphasizing the importance of developing formal and informal structures or processes of community engagement and designing local health action plans in cooperation with other administrations, such as regional and national governing bodies. To continue supporting these forms of local governance, we propose adapting the local administrations to the new social realities, with more horizontal and flexible organization models and resource allocation, integrating efficiency and evaluation processes, reclaiming local autonomy, and building local governance through networks and alliances

Extracellular Phosphorylation of TIMP-2 by Secreted c-Src Tyrosine Kinase Controls MMP-2 Activity.

The tissue inhibitor of metalloproteinases 2 (TIMP-2) is a specific endogenous inhibitor of matrix metalloproteinase 2 (MMP-2), which is a key enzyme that degrades the extracellular matrix and promotes tumor cell invasion. Although the TIMP-2:MMP-2 complex controls proteolysis, the signaling mechanism by which the two proteins associate in the extracellular space remains unidentified. Here we report that TIMP-2 is phosphorylated outside the cell by secreted c-Src tyrosine kinase. As a consequence, phosphorylation at Y90 significantly enhances TIMP-2 potency as an MMP-2 inhibitor and weakens the catalytic action of the active enzyme. TIMP-2 phosphorylation also appears to be essential for its interaction with the latent enzyme proMMP-2 in vivo. Absence of the kinase or non-phosphorylatable Y90 abolishes TIMP-2 binding to the latent enzyme, ultimately hampering proMMP-2 activation. Together, TIMP-2 phosphorylation by secreted c-Src represents a critical extracellular regulatory mechanism that controls the proteolytic function of MMP-2.

Intersectorality and social participation as coping policies for health inequities-worldwide / Intersectorialidad y participación social como políticas de afrontamiento para las desigualdades en salud en el mundo

Objective: To determine the impact that intersectoral policies and social participation, implemented worldwide, have had on the modification of the social determinants for health and on the reduction of social health inequities. Method: A scoping review of the literature published in the period 2005-2015 was performed. The literature search was conducted on PubMed and Scielo databases. Two researchers reviewed each document. Data were analysed according to the intersectoral action and social participation variables and according to the theoretical frameworks of the Social Determinants Model of the Commission on Social Determinants of Health (CSDH) and the theoretical constructs of Social Capital (SC) and Life Course (LC). Results: Out of 45 documents likely to be selected for final review, all of them based on title and abstract, 20 documents were eventually picked out and analysed; most them (n = 8) were conducted in all Latin America and Latin America's countries. Twelve documents reported intersectoral action associated with social participation in partnership with different institutions. Regarding theoretical frameworks, most of studies (n = 8) used CSDH and SC. In relation to health outcomes, the studies showed mainly: increased access to health and education, follow-up of pregnant women, increasing in prenatal examinations, reduction in malnutrition/child mortality, reduction in extreme poverty/hunger; reduction in epidemics/tuberculosis, control of alcohol/drug consumption, promotion of health/mental as well as basic sanitation improvements. Conclusions: Intersectoral and social participation experiences studied yielded positive outcomes regarding health status and quality of life in the communities in which such experiences were implemented

Determinar el impacto que las políticas intersectoriales y la participación social, implementadas en todo el mundo, han tenido tanto en la modificación de los determinantes sociales de la salud como en la reducción de las desigualdades sociales en salud. Método: Se realizó una revisión exploratoria (periodo 2005-2015). La búsqueda bibliográfica se hizo en las bases de datos PubMed y Scielo. Cada artículo fue revisado por dos investigadores. Los datos fueron analizados según las variables de acción intersectorial y participación social, y de acuerdo con los marcos teóricos: modelo de determinantes sociales de la Comisión de Determinantes Sociales de la Salud (CDSS) y constructos teóricos de capital social (CS) y curso de la vida. Resultados: De los 45 documentos seleccionables para la revisión final, basándose en el título y el resumen, 20 fueron seleccionados y analizados; la mayoría (n = 8) realizados en América Latina. Doce artículos informaban sobre acciones intersectoriales en asociación con participación social en colaboración con diferentes instituciones. En cuanto a los marcos teóricos, la mayoría (n = 8) utilizaron CDSS y CS. En relación con los resultados de salud, los estudios mostraron principalmente: mayor acceso a la salud y educación, seguimiento de embarazadas, aumento en exámenes prenatales, reducción de la desnutrición/mortalidad infantil, reducción de la pobreza extrema/hambre, reducción de las epidemias/tuberculosis, control del consumo de alcohol/drogas, promoción de la salud/salud mental, y mejoras en el saneamiento básico. Conclusiones: Las experiencias intersectoriales y de participación social estudiadas arrojan resultados positivos en el estado de salud y calidad de vida de las comunidades en que fueron implementadas

Intersectorality and social participation as coping policies for health inequities-worldwide.

OBJECTIVE: To determine the impact that intersectoral policies and social participation, implemented worldwide, have had on the modification of the social determinants for health and on the reduction of social health inequities. METHOD: A scoping review of the literature published in the period 2005-2015 was performed. The literature search was conducted on PubMed and Scielo databases. Two researchers reviewed each document. Data were analysed according to the intersectoral action and social participation variables and according to the theoretical frameworks of the Social Determinants Model of the Commission on Social Determinants of Health (CSDH) and the theoretical constructs of Social Capital (SC) and Life Course (LC). RESULTS: Out of 45 documents likely to be selected for final review, all of them based on title and abstract, 20 documents were eventually picked out and analysed; most them (n = 8) were conducted in all Latin America and Latin America's countries. Twelve documents reported intersectoral action associated with social participation in partnership with different institutions. Regarding theoretical frameworks, most of studies (n = 8) used CSDH and SC. In relation to health outcomes, the studies showed mainly: increased access to health and education, follow-up of pregnant women, increasing in prenatal examinations, reduction in malnutrition/child mortality, reduction in extreme poverty/hunger; reduction in epidemics/tuberculosis, control of alcohol/drug consumption, promotion of health/mental as well as basic sanitation improvements. CONCLUSIONS: Intersectoral and social participation experiences studied yielded positive outcomes regarding health status and quality of life in the communities in which such experiences were implemented.

New Approaches to Overcome Transport Related Drug Resistance in Trypanosomatid Parasites.

Leishmania and Trypanosoma are members of the Trypanosomatidae family that cause severe human infections such as leishmaniasis, Chagas disease, and sleeping sickness affecting millions of people worldwide. Despite efforts to eradicate them, migrations are expanding these infections to developing countries. There are no vaccines available and current treatments depend only on chemotherapy. Drug resistance is a major obstacle for the treatment of these diseases given that existing drugs are old and limited, with some having severe side effects. Most resistance mechanisms developed by these parasites are related with a decreased uptake or increased efflux of the drug due to mutations or altered expression of membrane transporters. Different new approaches have been elaborated that can overcome these mechanisms of resistance including the use of inhibitors of efflux pumps and drug carriers for both active and passive targeting. Here we review new formulations that have been successfully applied to circumvent resistance related to drug transporters, opening alternative ways to solve drug resistance in protozoan parasitic diseases.