Hypermigration of macrophages through the concerted action of GRA effectors on NF-κB/p38 signaling and host chromatin accessibility potentiates Toxoplasma dissemination.

Mononuclear phagocytes facilitate the dissemination of the obligate intracellular parasite Toxoplasma gondii. Here, we report how a set of secreted parasite effector proteins from dense granule organelles (GRA) orchestrates dendritic cell-like chemotactic and pro-inflammatory activation of parasitized macrophages. These effects enabled efficient dissemination of the type II T. gondii lineage, a highly prevalent genotype in humans. We identify novel functions for effectors GRA15 and GRA24 in promoting CCR7-mediated macrophage chemotaxis by acting on NF-κB and p38 MAPK signaling pathways, respectively, with contributions of GRA16/18 and counter-regulation by effector TEEGR. Further, GRA28 boosted chromatin accessibility and GRA15/24/NF-κB-dependent transcription at the Ccr7 gene locus in primary macrophages. In vivo, adoptively transferred macrophages infected with wild-type T. gondii outcompeted macrophages infected with a GRA15/24 double mutant in migrating to secondary organs in mice. The data show that T. gondii, rather than being passively shuttled, actively promotes its dissemination by inducing a finely regulated pro-migratory state in parasitized human and murine phagocytes via co-operating polymorphic GRA effectors.

Impact of climate risk materialization and ecological deterioration on house prices in Mar Menor, Spain.

The frequency and severity of extreme events related to climate change have intensified worldwide in the last decades. It is documented that increasing extreme rainfall and flooding cause more nutrient runoff into waterbodies, initiating numerous harmful algal bloom (HAB) events, especially in fragile ecosystems. We analyze the dramatic economic damage of one of these episodes in Mar Menor, the largest salt-water lagoon in Europe. We show that when the public perceived the severity of environmental degradation, the return on housing investment was 43% lower in the surroundings than in similar neighboring zones 6 years after the HAB (2015). This represents a loss in housing wealth of more than 4000 million euros, around ten times the gains of changing from dry-farming to irrigated crops, which makes this ecosystem fragile. Hence, we quantify some of the economic consequences of ecological deterioration linked to episodes of Global Climate Change.

Correction: Martínez-Valverde et al. Health Needs Assessment: Chronic Kidney Disease Secondary to Type 2 Diabetes Mellitus in a Population without Social Security, Mexico 2016-2032. Int. J. Environ. Res. Public Health 2022, 19, 9010.

In the original publication [...].

The Toxoplasma effector GRA28 promotes parasite dissemination by inducing dendritic cell-like migratory properties in infected macrophages.

Upon pathogen detection, macrophages normally stay sessile in tissues while dendritic cells (DCs) migrate to secondary lymphoid tissues. The obligate intracellular protozoan Toxoplasma gondii exploits the trafficking of mononuclear phagocytes for dissemination via unclear mechanisms. We report that, upon T. gondii infection, macrophages initiate the expression of transcription factors normally attributed to DCs, upregulate CCR7 expression with a chemotactic response, and perform systemic migration when adoptively transferred into mice. We show that parasite effector GRA28, released by the MYR1 secretory pathway, cooperates with host chromatin remodelers in the host cell nucleus to drive the chemotactic migration of parasitized macrophages. During in vivo challenge studies, bone marrow-derived macrophages infected with wild-type T. gondii outcompeted those challenged with MYR1- or GRA28-deficient strains in migrating and reaching secondary organs. This work reveals how an intracellular parasite hijacks chemotaxis in phagocytes and highlights a remarkable migratory plasticity in differentiated cells of the mononuclear phagocyte system.

In vitro evaluation of Resveratrol as a potential pre-exposure prophylactic drug against Trypanosoma cruzi infection.

Chagas' disease or American trypanosomiasis, caused by Trypanosoma cruzi infection, is an endemic disease in Latin America, which has spread worldwide in the past years. The drugs presently used for treatment have shown limited efficacy due to the appearance of resistant parasites and severe side effects. Some of the most recent studies on anti-parasitic drugs have been focused on protein acetylation, a reversible reaction modulated by Acetyl Transferases (KATs) and Deacetylases (KDACs). We have previously reported the anti-parasite activity of resveratrol (RSV), an activator of KDACs type III (or sirtuins), and showed that this drug can reduce the growth of T. cruzi epimastigotes and the infectivity of trypomastigotes. Since RSV is now widely used in humans due to its beneficial effects as an antioxidant, it has become an attractive candidate as a repurposing drug. In this context, the aim of the present study was to evaluate the ability of this drug to protect three different types of host cells from parasite infection. RSV treatment before parasite infection reduced the percentage of infected cells by 50-70% depending on the cell type. Although the mammalian cell lines tested showed different sensitivity to RSV, apoptosis was not significantly affected, showing that RSV was able to protect cells from infection without the activation of this process. Since autophagy has been described as a key process in parasite invasion, we also monitored this process on host cells pretreated with RSV. The results showed that, at the concentrations and incubation times tested, autophagy was not induced in any of the cell types evaluated. Our results show a partial protective effect of RSV in vitro, which justifies extending studies to an in vivo model to elucidate the mechanism by which this effect occurs.

Dosis efectivas asociadas a estudios híbridos SPECT-CT en pacientes adultos / Effective dose associated to hybrid SPECT-CT imaging in adult patients / Doses eficazes associadas a estudos de SPECT-CT híbridos em pacientes adultos

Introducción: la técnica de imagen híbrida de SPECT-CT combina la imagen de la tomografía por emisión de fotón único (SPECT) con el estudio de tomografía computada (TC), obteniendo información funcional y anatómica en un mismo estudio. La dosis efectiva total de radiación ionizante recibida en los estudios SPECT-CT puede ser estimada a partir de la dosis efectiva atribuible a la actividad administrada del radiofármaco y la dosis efectiva del componente de tomografía computada (TC). Objetivos: estimar la dosis efectiva total en los protocolos SPECT-CT utilizados en población adulta y determinar el aporte adicional del estudio TC sobre la dosis efectiva total. Método: se evaluaron 258 estudios SPECT-CT para estimar la dosis efectiva total aportada por la administración de los radiofármacos y los estudios de TC de baja dosis. Para estimar el aporte de ambos componentes se utilizaron factores de conversión específicos de cada radiofármaco y región explorada mediante TC. Resultados: la dosis efectiva total (media ± DS) en los estudios SPECT-CT fueron: 12,4 ± 1,44 mSv en el estudio de perfusión miocárdica, 1,14 ± 0,25 mSv en ganglio centinela de mama, 8,6 ± 0,6 mSv paratiroides, 1,48 ± 1,02 mSv tiroides y los estudios óseos de las regiones de cuello 4,5 ± 0,3, tórax 6,07 ± 0,3 mSv, abdomen y pelvis 6,1 ± 0,3 mSv. La dosis de radiación aportada por el estudio TC se encuentra entre 0,46 mSv para la región del tórax en el estudio de ganglio centinela de mama y 2,3 mSv para el SPECT-CT óseo en la región de abdomen y pelvis. Conclusión: se logró estimar la dosis efectiva en los protocolos SPECT-CT de uso clínico más frecuente en población adulta y el aporte de los estudios TC a la dosis efectiva total siendo relativamente baja comparado con la dosis aportada por los radiofármacos administrados con la excepción del estudio de ganglio centinela donde la contribución del componente TC es aproximadamente la mitad de la dosis efectiva total.

Introduction: SPECT-CT Hybrid image technique combines the SPECT (single-photon emission computed tomography) image with the CT (computerized tomography) image to obtain both functional and anatomical images in the same study. The total effective ionizing radiation dose received in SPECT-CT studies may be estimated based on the effective dose from the radiopharmaceutical administered and the effective dose from the CT (computerized tomography) component. Objectives: the study aims to estimate the total effective dose in SPECT-CT protocols applied for the adult population, and to determine the additional contribution from the CT component to the total effective dose. Method: 258 SPECT-CT studies were evaluated to estimate the total effective dose from the administration of radiopharmaceuticals and low dose CT studies. Specific conversion factors for each radiopharmaceutical and area of the body explored with the CT were used to estimate radiation doses from both components. Results: total effective dose (average ± SD) in the SPECT-CT studies was: 12.4 ± 1.44 mSv in the myocardial perfusion study, 1.14 ± 0.25 mSv in the breast sentinel lymph node study, 8.6 ± 0.6 mSv in the parathyroid study, 1.48 ± 1.02 mSv in the thyroid study. As to bone studies, doses found were: 4.5 ± 0.3, in neck studies, 6.07 ± 0.3 mSv in thoracic studies and 6.1 ± 0.3 mSv in abdominal and pelvic studies. The radiation dose from the CT study ranges from 0.46 mSv for the thoracic region on the breast sentinel lymph node study to 2.3 mSv for the bone SPECT-CT study of the abdominal and pelvic region. Conclusions: we managed to estimate the effective dose in the the most frequently used SPECT-CT protocols for the adult population and the contribution of CT studies to the total effective dose. It was found to be relatively low when compared to the dose contributed by the radiopharmaceuticals administered, with the exception of the sentinel lymph node study for which the contribution from the CT study is approximately half the total effective dose.

Introdução: a técnica de imagem híbrida SPECT-CT combina a imagem de tomografia por emissão de fóton único (SPECT) com o estudo de tomografia computadorizada (TC), obtendo informações funcionais e anatômicas no mesmo estudo. A dose efetiva total de radiação ionizante recebida em estudos SPECT-CT pode ser estimada a partir da dose efetiva atribuível à atividade administrada do radiofármaco e da dose efetiva do componente de tomografia computadorizada (TC). Objetivos: estimar a dose efetiva total nos protocolos SPECT-CT utilizados na população adulta e determinar a contribuição adicional do estudo de TC na dose efetiva total. Método : 258 estudos SPECT-CT foram avaliados para estimar a dose efetiva total fornecida pela administração de radiofármacos e estudos de TC de baixa dose. Para estimar a contribuição de ambos os componentes, foram utilizados fatores de conversão específicos para cada radiofármaco e região explorada pela TC. ⁠ Resultados: a dose efetiva total (média ± DP) nos estudos SPECT-CT foi: 12,4 ± 1,44 mSv no estudo de perfusão miocárdica, 1,14 ± 0,25 mSv no linfonodo sentinela mamário, 8,6 ± 0,6 mSv paratireoide, 1,48 ± 1,02 mSv estudos de tireoide e ossos das regiões do pescoço 4,5 ± 0,3, tórax 6,07 ± 0,3 mSv, abdômen e pelve 6,1 ±0,3mSv. A dose de radiação fornecida pelo estudo de TC está entre 0,46 mSv para a região do tórax no estudo do linfonodo sentinela da mama e 2,3 mSv para o SPECT-CT ósseo na região do abdome e pelve. Conclusão: foi possível estimar a dose efetiva nos protocolos de SPECT-CT mais utilizados clinicamente na população adulta e a contribuição dos estudos de TC para a dose efetiva total, sendo relativamente baixa em relação à dose fornecida pelos radiofármacos administrados com a exceção do estudo do linfonodo sentinela onde a contribuição do componente TC é aproximadamente metade da dose efetiva total.

Biological features of TcM: A new Trypanosoma cruzi isolate from Argentina classified into TcV lineage.

Trypanosoma cruzi, the etiologic agent of Chagas disease (CD) presents a wide genetic and phenotypic diversity that is classified into seven lineages or discrete typing units (DTU: TcI to TcVI and Tcbat). Although isolates and strains that belong to a particular group can share some attributes, such as geographic distribution, others like growth rate, cell tropism, and response to treatment can be highly variable. In addition, studies that test new trypanocidal drugs are frequently conducted on T. cruzi strains maintained for a long time in axenic culture, resulting in changes in parasite virulence and other important features. This work aimed to isolate and characterize a new T. cruzi strain from a chronic Chagas disease patient. The behavior of this isolate was studied by using standard in vitro assays and in vivo mice infection tests and compared with the T. cruzi Y strain (TcY), broadly used in research laboratories worldwide. Data showed that TcM behaves as a slow-growing strain in vitro that develops chronic infections in mice and displays high tropism to muscular tissues, in accordance with its clinical performance. In contrast, the Y strain behaved as an acute strain that can infect different types of cells and tissues. Interestingly, TcM, which belongs to DTU TcV, is more susceptible to benznidazole than TcY, a TcII strain considered moderately resistant to this drug. These differential properties contribute to the characterization of a TcV strain, one of the main lineages in the southern countries of South America, and open the possibility to introduce changes that improve the management of Chagas patients in the future.

Health Needs Assessment: Chronic Kidney Disease Secondary to Type 2 Diabetes Mellitus in a Population without Social Security, Mexico 2016-2032.

Health needs assessment is a relevant tracer of planning process of healthcare programs. The objective is to assess the health needs of chronic kidney disease (CKD) secondary to type 2 diabetes mellitus (T2 DM) in a population without social security in Mexico. The study design was a statistical simulation model based on data at the national level of Mexico. A stochastic Markov model was used to simulate the progression from diabetes to CKD. The time horizon was 16 years. The results indicate that in 2022, kidney damage progression and affectation in the diabetic patient cohort will be 34.15% based on the time since T2 DM diagnosis. At the end of the 16-year period, assuming that the model of care remains unchanged, early renal involvement will affect slightly more than twice as many patients (118%) and cases with macroalbuminuria will triple (228%). The need for renal replacement therapy will more than double (169%). Meanwhile, deaths associated with cardiovascular risk will more than triple (284%). We concluded that the clinical manifestations of patients with CKD secondary to T2 DM without social security constitute a double challenge. The first refers to the fact that the greatest health need is early care of CKD, and the second is the urgent need to address cardiovascular risk in order to reduce deaths in the population at risk.

Mobilome of Apicomplexa Parasites.

Transposable elements (TEs) are mobile genetic elements found in the majority of eukaryotic genomes. Genomic studies of protozoan parasites from the phylum Apicomplexa have only reported a handful of TEs in some species and a complete absence in others. Here, we studied sixty-four Apicomplexa genomes available in public databases, using a 'de novo' approach to build candidate TE models and multiple strategies from known TE sequence databases, pattern recognition of TEs, and protein domain databases, to identify possible TEs. We offer an insight into the distribution and the type of TEs that are present in these genomes, aiming to shed some light on the process of gains and losses of TEs in this phylum. We found that TEs comprise a very small portion in these genomes compared to other organisms, and in many cases, there are no apparent traces of TEs. We were able to build and classify 151 models from the TE consensus sequences obtained with RepeatModeler, 96 LTR TEs with LTRpred, and 44 LINE TEs with MGEScan. We found LTR Gypsy-like TEs in Eimeria, Gregarines, Haemoproteus, and Plasmodium genera. Additionally, we described LINE-like TEs in some species from the genera Babesia and Theileria. Finally, we confirmed the absence of TEs in the genus Cryptosporidium. Interestingly, Apicomplexa seem to be devoid of Class II transposons.

Software evaluation for de novo detection of transposons.

Transposable elements (TEs) are major genomic components in most eukaryotic genomes and play an important role in genome evolution. However, despite their relevance the identification of TEs is not an easy task and a number of tools were developed to tackle this problem. To better understand how they perform, we tested several widely used tools for de novo TE detection and compared their performance on both simulated data and well curated genomic sequences. As expected, tools that build TE-models performed better than k-mer counting ones, with RepeatModeler beating competitors in most datasets. However, there is a tendency for most tools to identify TE-regions in a fragmented manner and it is also frequent that small TEs or fragmented TEs are not detected. Consequently, the identification of TEs is still a challenging endeavor and it requires a significant manual curation by an experienced expert. The results will be helpful for identifying common issues associated with TE-annotation and for evaluating how comparable are the results obtained with different tools.

From telomere to telomere: The transcriptional and epigenetic state of human repeat elements.

Mobile elements and repetitive genomic regions are sources of lineage-specific genomic innovation and uniquely fingerprint individual genomes. Comprehensive analyses of such repeat elements, including those found in more complex regions of the genome, require a complete, linear genome assembly. We present a de novo repeat discovery and annotation of the T2T-CHM13 human reference genome. We identified previously unknown satellite arrays, expanded the catalog of variants and families for repeats and mobile elements, characterized classes of complex composite repeats, and located retroelement transduction events. We detected nascent transcription and delineated CpG methylation profiles to define the structure of transcriptionally active retroelements in humans, including those in centromeres. These data expand our insight into the diversity, distribution, and evolution of repetitive regions that have shaped the human genome.

Different kinetoplast degradation patterns in American Trypanosoma vivax strains: Multiple independent origins or fast evolution?

We analyzed the kinetoplast (mitochondrial genome) of Trypanosoma vivax strains from America and Africa to determine their precise architecture and to understand their adaptive response to mechanical transmission. The use of long-read based assemblies that retain individuality of tandem repeats, without erasing inter-copy variability, allowed us to investigate the evolutionary dynamics of repetitive kinetoplast-DNA. This analysis revealed that repeat elements located in edges of repeat clusters are less active in terms of renewal, whereas internal copies appear to undergo a permanent process of birth-and-death. Comparing different American strains with the African Y486 strain, we found that in the former, protein coding genes from the maxicircle contain several function disrupting mutations that with very few exceptions are present in one or the other American strain but not in both, suggesting the absence of common ancestry for most of the genomic changes that led to their loss of oxidative phosphorylation capacity. Analysis of another component of kinetoplast, the minicircles, revealed great loss of diversity, and loss of their encoded guideRNAs. Both groups of American strains retain minimal sets required to edit the still functional A6-APTase and RPS12 genes. The extensive maxi- and minicircle divergence suggests a history of multiple introduction events in America of strains that probably started to degrade their kinetoplast in Africa. The notion that kinetoplast degradation began after incursion in America would imply a pace of accumulation of genetic changes considerably faster than other trypanosomatids.

The Trypanosoma cruzi TcTASV-C protein subfamily administrated with U-Omp19 promotes a protective response against a lethal challenge in mice.

The development of a Chagas disease vaccine has yet the need for the identification of novel combinations of antigens and adjuvants. Here, the performance of TcTASV-C proteins that are virulence factors of trypomastigotes and belong to a novel surface protein family specific for T. cruzi, have been evaluated as antigens for a prophylactic vaccine. Several immunization schemes in which TcTASV-C was combined with aluminum hydroxide, saponin and/or U-Omp19 were assayed. Aluminum hydroxide and saponin were assayed together to trigger different pathways of the immune response simultaneously. U-Omp19 is a promising novel adjuvant able to promote a Th1 immune response with IFNg production, thus an interesting molecule to be tested as adjuvant for the control of T. cruzi infection. Therefore, U-Omp19 was added to the aluminum hydroxide-saponin formulation as well as assayed individually with TcTASV-C. The immunization with TcTASV-C and U-Omp19 had the best performance as a prophylactic vaccine. Mice presented the lowest parasitemias and improved survival by 40% after being challenged with a highly virulent T. cruzi strain, which promoted 100% mortality in all other immunized groups. Immunization with TcTASV-C and U-Omp19 triggered cellular responses with IFN-γ and IL-17 production and with lytic antibodies that could explain the protection achieved by this vaccination scheme. To our knowledge, this is the first time that U-Omp19 is tested with a defined T. cruzi antigen in a vaccine formulation.

Transmigration of Trypanosoma cruzi trypomastigotes through 3D cultures resembling a physiological environment.

To disseminate and colonise tissues in the mammalian host, Trypanosoma cruzi trypomastogotes should cross several biological barriers. How this process occurs or its impact in the outcome of the disease is largely speculative. We examined the in vitro transmigration of trypomastigotes through three-dimensional cultures (spheroids) to understand the tissular dissemination of different T. cruzi strains. Virulent strains were highly invasive: trypomastigotes deeply transmigrate up to 50 µm inside spheroids and were evenly distributed at the spheroid surface. Parasites inside spheroids were systematically observed in the space between cells suggesting a paracellular route of transmigration. On the contrary, poorly virulent strains presented a weak migratory capacity and remained in the external layers of spheroids with a patch-like distribution pattern. The invasiveness-understood as the ability to transmigrate deep into spheroids-was not a transferable feature between strains, neither by soluble or secreted factors nor by co-cultivation of trypomastigotes from invasive and non-invasive strains. Besides, we demonstrated that T. cruzi isolates from children that were born congenitally infected presented a highly migrant phenotype while an isolate from an infected mother (that never transmitted the infection to any of her children) presented significantly less migration. In brief, we demonstrated that in a 3D microenvironment each strain presents a characteristic migration pattern that can be associated to their in vivo behaviour. Altogether, data presented here repositionate spheroids as a valuable tool to study host-pathogen interactions.

Cáncer de mama y su respuesta al tratamiento neoadyuvante según subtipo molecular / Breast cancer and its response to neoadjuvant treatment according to molecular subtipe

El cáncer de mama es una de las patologías más frecuentes a nivel mundial y en el Ecuador ocupa un sitio importante dentro de la mortalidad; en pacientes con tumores de estadios avanzados la quimioterapia neodyuvante es el procedimiento indicado para lograr una reducción tumoral satisfactoria. El objetivo fue determinar la respuesta clínica y patológica en pacientes con cáncer de mama tratadas con quimioterapia neoadyuvante según cada subtipo molecular, atendidos en el hospital "Teodoro Maldonado Carbo" en el período 2015 a 2017. Se hizo uso de un diseño no experimental, transversal de tipo correlacional. Pacientes con cáncer de mama que recibieron neoadyuvancia, en su mayoría con quimioterapia basada en antraciclinas y taxanos. Se clasificó a las pacientes por sus subtipos moleculares, los mismos se obtuvieron en base a las características inmunohistoquímicas de los reportes de patología que constan en el sistema AS-400. Se comprobó la respuesta clínica al tratamiento usando los Criterios RECIST 1.1. Como resultado los 171 pacientes fueron analizados. La edad promedio de las pacientes fue 55 13 años de edad; el 25% fueron luminal B (HER+), 24% luminal B (HER-), 22% triple negativo, 18% HER2+ y 12% luminal A; el 52% de las pacientes tuvieron estadio III de la enfermedad; el 75% (129) de las pacientes fue realizada una mastectomía radical modificada. Se pudo concluir que la respuesta patológica completa en pacientes con tratamiento neoadyuvante se relaciona con los subtipos moleculares y esto es estadísticamente significativo. Además, se evidenció las mayores tasas de respuesta patológica completa en los grupos moleculares de HER2+ y triple negativo.

Breast cancer is one of the most frequent pathologies worldwide and in Ecuador it occupies an important place in mortality. In patients with advanced stage tumors, the neo-adjuvant chemotherapy is the indicated procedure to achieve a satisfactory tumor reduction. The aim was to determine the clinical and pathological response in patients with breast cancer treated with neoadjuvant chemotherapy according to each molecular subtype, treated at the "Teodoro Maldonado Carbo" hospital in the period 2015 to 2017. We used a non-experimental, crosssectional type design. Patients with breast cancer who received neoadjuvant, mostly with chemotherapy based on anthracyclines and taxanes. The patients were classified by their molecular subtypes, they were obtained based on the immunohistochemical characteristics of the pathology reports that appear in the AS-400 system. The clinical response to treatment was checked using the RECIST 1.1 Criteria. As a result, a sum of 171 patients were analyzed. The average age of the patients was 55 + 13 years old; 25% were luminal B (Her +), 24% luminal B (Her-), 22% triple negative, 18% Her2 + and 12% luminal A; 52% of the patients had stage III of the disease; 75% (129) of the patients underwent a modified radical mastectomy. As a conclusion, the complete pathological response in patients with neoadjuvant treatment is related to molecular subtypes and this is statistically significant. Also, the highest rates of complete pathological response in the molecular groups of Her2 + and triple negative were evident.

Human papillomavirus type 18 E5 oncoprotein cooperates with E6 and E7 in promoting cell viability and invasion and in modulating the cellular redox state

BACKGROUND High-risk human papillomaviruses (HR-HPVs) are the etiological agents of cervical cancer. Among them, types 16 and 18 are the most prevalent worldwide. The HPV genome encodes three oncoproteins (E5, E6, and E7) that possess a high transformation potential in culture cells when transduced simultaneously. In the present study, we analysed how these oncoproteins cooperate to boost key cancer cell features such as uncontrolled cell proliferation, invasion potential, and cellular redox state imbalance. Oxidative stress is known to contribute to the carcinogenic process, as reactive oxygen species (ROS) constitute a potentially harmful by-product of many cellular reactions, and an efficient clearance mechanism is therefore required. Cells infected with HR-HPVs can adapt to oxidative stress conditions by upregulating the formation of endogenous antioxidants such as catalase, glutathione (GSH), and peroxiredoxin (PRX). OBJECTIVES The primary aim of this work was to study how these oncoproteins cooperate to promote the development of certain cancer cell features such as uncontrolled cell proliferation, invasion potential, and oxidative stress that are known to aid in the carcinogenic process. METHODS To perform this study, we generated three different HaCaT cell lines using retroviral transduction that stably expressed combinations of HPV-18 oncogenes that included HaCaT E5-18, HaCaT E6/E7-18, and HaCaT E5/E6/E7-18. FINDINGS Our results revealed a statistically significant increment in cell viability as measured by MTT assay, cell proliferation, and invasion assays in the cell line containing the three viral oncogenes. Additionally, we observed that cells expressing HPV-18 E5/E6/E7 exhibited a decrease in catalase activity and a significant augmentation of GSH and PRX1 levels relative to those of E5, E6/E7, and HaCaT cells. MAIN CONCLUSIONS This study demonstrates for the first time that HPV-18 E5, E6, and E7 oncoproteins can cooperate to enhance malignant transformation.

Fístula arteriovenosa dural espinal: a propósito de un caso / Spinal dural arteriovenous fistula: a case report

Las fístulas arteriovenosas durales medulares son malformaciones vasculares adquiridas que constituyen una causa muy infrecuente de mielopatía progresiva (5-10 casos por millón de habitantes por año). La resonancia magnética es el estudio por imágenes de elección para su diagnóstico. A continuación presentamos el caso de una paciente femenina de 89 años, que consultó a la guardia de nuestra institución por un cuadro de paraparesia moderada asociada a parestesias e incontinencia urinaria posterior a esfuerzo físico. Se le diagnosticó una fístula arteriovenosa dural medular como causante de su cuadro. (AU)

Spinal dural arteriovenous fistulas (SDAVF) are acquired spinal vascular malformations and a rare cause of progressive myelopathy (5-10 new cases per year and per 1 million inhabitants). Magnetic resonance imaging is the diagnosis modality of choice. We present a case of a 89-year-old female patient who consulted the emergency department of our institution because of paraparesis and lower extremities paresthesias associated with urinary incontinence post physical effort. With the final diagnosis of spinal dural arteriovenous fistula, as a cause of the clinical symptoms. (AU)

Transmigration of Trypanosoma cruzi Trypomastigotes through 3D Spheroids Mimicking Host Tissues.

While cellular invasion by T. cruzi trypomastigotes and intracellular amastigote replication are well-characterized events that have been described by using 2D monolayer cultures, other relevant parasite-host interactions, like the dynamics of tissue invasiveness, cannot be captured using monolayer cultures. Spheroids constitute a valuable three-dimensional (3D) culture system because they mimic the microarchitecture of tissues and provide an environment similar to the encountered in natural infections, which includes the presence of extracellular matrix as well as 3D cell-cell interactions. In this work, we describe a protocol for studying transmigration of T. cruzi trypomastigotes into 3D spheroids. In the experimental setup, cells and parasites are labelled with two fluorescent dyes, allowing their visualization by confocal microscopy. We also describe the general procedure and setting of the confocal microscope and downstream applications for acquisition and reconstruction of 3D images. This model was employed to analyze the transmigration of trypomastigotes from the highly virulent and pantropic RA T. cruzi strain. Of course, other aspects encountered by T. cruzi in the mammalian host environment can be studied with this methodology.