Oportunidades perdidas en la detección temprana de la infección por VIH en pacientes con infecciones de transmisión sexual: un estudio de vida real / Missed opportunities in the early detection of HIV infection in patients with sexually transmitted infections: A real-life study

Introducción: Las enfermedades de transmisión sexual, como la cervicitis, la proctitis y la uretritis, se asocian a altas tasas de infección por VIH. Ante la sospecha de estas patologías, se debería solicitar una serología del VIH. Material y métodos: Estudio retrospectivo realizado durante 2018 en el Hospital Costa del Sol (Marbella, Málaga). Se revisaron las serologías para el VIH solicitadas en pacientes a los que se les pidió una PCR para Chlamydia trachomatis y Neisseria gonorrhoeae. Resultados: Se valoraron 1.818 pacientes, en los que se realizó serología para el VIH al 44,7%, de las cuales 14 (1,7%) resultaron positivas. El 55,3% restante fueron oportunidades perdidas de diagnóstico. Conclusiones: Las infecciones por C.trachomatis y N.gonorrhoeae están asociadas a una elevada tasa de infección oculta por el VIH. El grado de sospecha de VIH en esta población sigue siendo bajo, y resulta esencial que se refuerce ante la posibilidad de infección por estas patologías.(AU)

Introduction: Sexually transmitted diseases such as cervicitis, proctitis and urethritis are associated with high rates of HIV infection. When these pathologies are suspected, HIV serology should be requested. Material and methods: A Retrospective study was performed during 2018 at the Hospital Costa del Sol (Marbella, Málaga, Spain). HIV serologies requested in patients who were asked for PCR for Chlamydia trachomatis and Neisseria gonorrhoeae were reviewed. Results: A total of 1818 patients were evaluated, in which HIV serology was performed in 44.7%, of which 14 (1.7%) were positive. The remaining 55.3% were missed diagnostic opportunities. Conclusions: C.trachomatis and N.gonorrhoeae infections are associated with a high rate of occult HIV infection. The degree of suspicion of HIV in this population remains low and it is essential that it be reinforced in the presence of the possibility of infection by these pathologies.(AU)

Nonsurgical Management of Shoulder Pain in Rotator Cuff Tears: Ultrasound-Guided Biceps Tenotomy Combined With Corticosteroid Injection.

Traditionally, the management of rotator cuff tears in elderly individuals has involved surgical intervention, specifically biceps tenotomy. However, surgical procedures come with inherent medical risks and significant financial costs. As an alternative, ultrasound-guided biceps tenotomy combined with corticosteroid has emerged as a highly effective, well-tolerated, and cost-efficient option. This article aims to describe the procedural technique of nonsurgical management using ultrasound-guided biceps tenotomy combined with corticosteroid injection.

Missed opportunities in the early detection of HIV infection in patients with sexually transmitted infections: A real-life study.

INTRODUCTION: Sexually transmitted diseases such as cervicitis, proctitis and urethritis are associated with high rates of HIV infection. When these pathologies are suspected, HIV serology should be requested. MATERIAL AND METHODS: A retrospective study was performed during 2018 at the Hospital Costa del Sol (Marbella, Málaga, Spain). HIV serologies requested in patients who were asked for PCR for Chlamydia trachomatis and Neisseria gonorrhoeae were reviewed. RESULTS: A total of 1818 patients were evaluated, in which HIV serology was performed in 44.7%, of which 14 (1.7%) were positive. The remaining 55.3% were missed diagnostic opportunities. CONCLUSIONS: C. trachomatis and N. gonorrhoeae infections are associated with a high rate of occult HIV infection. The degree of suspicion of HIV in this population remains low and it is essential that it be reinforced in the presence of the possibility of infection by these pathologies.

Taxonomic novelties in Haplopappus (Asteraceae, Astereae) from Chile.

Two new species of Haplopappus (Asteraceae) from central Chile are described in this article. Haplopappuscolliguayensissp. nov. is restricted to La Chapa hill, Colliguay, Valparaíso Region, and is most similar to H.undulatus but differs from the latter in its stem indumentum, leaf shape and margin, and synflorescence arrangement. Haplopappusteillierisp. nov. has been recorded from four high-Andean localities in the Choapa, Petorca, Rocín and Aconcagua river basins, and is most similar to H.punctatus but differs from the latter in its leaf length and margin, number of peduncles per twig, width of outer phyllaries, number of ray florets per capitulum, and achene dimensions. Additionally, we propose the reinstatement of H.kingii in agreement with an exhaustive revision of type material and protologues, as well as the study of herbarium specimens. Haplopappuskingii is restricted to mountainous areas in the southern portion of the Atacama Region, and resembles H.parvifolius and H.retinervius but differs from both by its leaf margin and apex, besides additional differences from each. We provide morphological descriptions, field images, distributional maps, conservation assessments, and taxonomic notes for the three species treated, as well as illustrations for the novel taxa.

Evolutionary-Related High- and Low-Virulent Classical Swine Fever Virus Isolates Reveal Viral Determinants of Virulence.

Classical swine fever (CSF) has been eradicated from Western and Central Europe but remains endemic in parts of Central and South America, Asia, and the Caribbean. CSF virus (CSFV) has been endemic in Cuba since 1993, most likely following an escape of the highly virulent Margarita/1958 strain. In recent years, chronic and persistent infections with low-virulent CSFV have been observed. Amino acid substitutions located in immunodominant epitopes of the envelope glycoprotein E2 of the attenuated isolates were attributed to positive selection due to suboptimal vaccination and control. To obtain a complete picture of the mutations involved in attenuation, we applied forward and reverse genetics using the evolutionary-related low-virulent CSFV/Pinar del Rio (CSF1058)/2010 (PdR) and highly virulent Margarita/1958 isolates. Sequence comparison of the two viruses recovered from experimental infections in pigs revealed 40 amino acid differences. Interestingly, the amino acid substitutions clustered in E2 and the NS5A and NS5B proteins. A long poly-uridine sequence was identified previously in the 3' untranslated region (UTR) of PdR. We constructed functional cDNA clones of the PdR and Margarita strains and generated eight recombinant viruses by introducing single or multiple gene fragments from Margarita into the PdR backbone. All chimeric viruses had comparable replication characteristics in porcine monocyte-derived macrophages. Recombinant PdR viruses carrying either E2 or NS5A/NS5B of Margarita, with 36 or 5 uridines in the 3'UTR, remained low virulent in 3-month-old pigs. The combination of these elements recovered the high-virulent Margarita phenotype. These results show that CSFV evolution towards attenuated variants in the field involved mutations in both structural and non-structural proteins and the UTRs, which act synergistically to determine virulence.

A representação das mulheres na publicidade exterior e nas intervenções urbanas: um estudo comparativo, interseccional e multimodal entre as cidades de natal, recife (brasil) e barcelona (espanha)

Neste artigo objetivamos estudar as imagens de mulheres na publicidade exterior e nas intervenções urbanas, perceber os discursos recorrentes, fenotipos e identificar as ausências e presenças de marcadores sociais da diferença. Observamos as cidades brasileiras de Natal (Rio Grande do Norte) e Recife (Pernambuco), em comparação com a cidade espanhola de Barcelona (Catalunha). Utilizamos a etnografia para o mapeamento e coleta dos dados, cujo período corresponde a abril de 2021 a novembro de 2022. Adotamos parte da metodologia multimodal (análise da emissão) e recorremos à perspectiva interseccional para refletir criticamente sobre as imagens. Concluímos que as intervenções urbanas apresentam temas emergentes do universo das mulheres, como o feminicídio e o racismo, e direcionam à educação feminista, ou seja, elas surgem como contraponto discursivo às imagens de submissão de mulheres propagadas pela publicidade.

En este artículo nos propusimos estudiar las imágenes de las mujeres en la publicidad exterior y las intervenciones urbanas, percibir los discursos recurrentes, los fenotipos e identificar las ausencias y presencias de marcadores sociales de diferencia. Observamos las ciudades brasileñas de Natal (Rio Grande do Norte) y Recife (Pernambuco), en comparación con la ciudad española de Barcelona (Cataluña). Utilizamos la etnografía para el mapeo y la recopilación de datos, cuyo período corresponde a abril de 2021 a noviembre de 2022. Adoptamos parte de la metodología multimodal (análisis de emisiones) y recurrimos a la perspectiva interseccional para reflexionar críticamente sobre las imágenes. Concluimos que las intervenciones urbanas presentan temas emergentes del universo femenino, como el feminicidio y el racismo, y apuntan a la educación feminista, es decir, surgen como contrapunto discursivo a las imágenes de sumisión de la mujer propagadas por la publicidad.

Unraveling biomarkers of exposure for tenuazonic acid through urinary metabolomics.

Mycotoxins are secondary metabolites produced by fungi such as Aspergillus, Alternaria, and Penicillium, affecting nearly 80% of global food crops. Tenuazonic acid (TeA) is the major mycotoxin produced by Alternaria alternata, a prevalent pathogen affecting plants, fruits, and vegetables. TeA is notably prevalent in European diets, however, TeA biomarkers of exposure and metabolites remain unknown. This research aims to bridge this knowledge-gap by gaining insights about human TeA exposure and metabolization. Nine subjects were divided into two groups. The first group received a single bolus of TeA at the Threshold of Toxicological Concern (TTC) to investigate the presence of TeA urinary biomarkers, while the second group served as a control. Sixty-nine urinary samples were prepared and analyzed using UPLC-Xevo TQ-XS for TeA quantification and UPLC-Orbitrap Exploris for polar metabolome acquisition. TeA was rapidly excreted during the first 13 h and the fraction extracted was 0.39 ± 0.22. The polar metabolome compounds effectively discriminating the two groups were filtered using Orthogonal Partial Least Squares-Discriminant Analysis and subsequently annotated (n = 122) at confidence level 4. Finally, the urinary metabolome was compared to in silico predicted TeA metabolites. Nine metabolites, including oxidized, N-alkylated, desaturated, glucuronidated, and sulfonated forms of TeA were detected.

The autophagy protein Def8 is altered in Alzheimer's disease and Aß42-expressing Drosophila brains.

Alzheimer's disease (AD) is the most common neurodegenerative disorder, characterized by protein accumulation in the brain as a main neuropathological hallmark. Among them, Aß42 peptides tend to aggregate and create oligomers and plaques. Macroautophagy, a form of autophagy characterized by a double-membrane vesicle, plays a crucial role in maintaining neuronal homeostasis by degrading protein aggregates and dysfunctional organelles as a quality control process. Recently, DEF8, a relatively uncharacterized protein, has been proposed as a participant in vesicular traffic and autophagy pathways. We have reported increased DEF8 levels in lymphocytes from mild cognitive impairment (MCI) and early-stage AD patients and a neuronal profile in a murine transgenic AD model. Here, we analyzed DEF8 localization and levels in the postmortem frontal cortex of AD patients, finding increased levels compared to healthy controls. To evaluate the potential function of DEF8 in the nervous system, we performed an in silico assessment of its expression and network profiles, followed by an in vivo evaluation of a neuronal Def8 deficient model using a Drosophila melanogaster model of AD based on Aß42 expression. Our findings show that DEF8 is an essential protein for maintaining cellular homeostasis in the nervous system, and it is upregulated under stress conditions generated by Aß42 aggregation. This study suggests DEF8 as a novel actor in the physiopathology of AD, and its exploration may lead to new treatment avenues.

NS1 and PA-X of H1N1/09 influenza virus act in a concerted manner to manipulate the innate immune response of porcine respiratory epithelial cells.

Live-attenuated influenza A viruses (LAIV) may be superior to inactivated or subunit vaccines since they can be administered via mucosal routes to induce local immunity in the respiratory tract. In addition, LAIV are expected to trigger stronger T-cell responses that may protect against a broader range of antigen-drifted viruses. However, the development of LAIV is challenging since a proper balance between immunogenicity and safety has to be reached. In this study, we took advantage of reverse genetics to generate three LAIV based on the pandemic H1N1 2009 (pH1N1/09) virus strain: ΔPA-X, which is defective in the synthesis of the accessory PA-X protein, NS1(1-126) lacking 93 amino acids at the C-terminus of the NS1 protein, and a combination of both. Characterization of these recombinant viruses using a novel porcine bronchiolar epithelial cell line (T3) revealed that the ΔPA-X mutant replicated similar to wild type (WT) virus. However, in contrast to the parental virus the ΔPA-X mutant allowed transcription of genes involved in cell cycle progression and limits apoptosis. The NS1(1-126) mutant also replicated comparable to WT virus, but triggered the release of type I and III IFN and several chemokines and cytokines. Surprisingly, only the NS1(1-126)/ΔPA-X double mutant was significantly attenuated on T3 cells, and this was associated with enhanced transcription of genes of the innate immune system and complete absence of apoptosis induction. In conclusion, these findings indicate that NS1 and PA-X act in a concerted manner to manipulate the host cell response, which may help to develop swine LAIV vaccine with a more favorable balance of safety and immunogenicity.

Effects of a Contextual Adaptation of the Unified Protocol in Multiple Emotional Disorders in Individuals Exposed to Armed Conflict in Colombia: A Randomized Clinical Trial.

Importance: A transdiagnostic treatment, the Unified Protocol, is as effective as single diagnostic protocols in comorbid emotional disorders in clinical populations. However, its effects on posttraumatic stress disorder and other emotional disorders in individuals living in war and armed conflict contexts have not been studied. Objective: To evaluate the efficacy of a cultural and contextual adaptation of the Unified Protocol (CXA-UP) on posttraumatic stress disorder, anxiety, and depression compared to waitlist control in individuals exposed to armed conflict in Colombia. Design, Setting, and Participants: From April 2017 to March 2020, 200 participants 18 years and older were randomly assigned to the CXA-UP or to a waitlist condition. CXA-UP consisted of 12 to 14 twice-a-week or weekly individual 90-minute face-to-face sessions. Outcomes were assessed at baseline, posttreatment, and 3 months following treatment. Analyses were performed and compared for all randomly allocated participants (intent-to-treat [ITT]) and for participants who completed all sessions and posttreatment measures (per protocol [PP]). The study took place at an outpatient university center and included individuals who were registered in the Colombian Victims Unit meeting DSM-5 diagnostic criteria for posttraumatic stress disorder, anxiety, or depression or were severely impaired by anxiety or depression. Individuals who were receiving psychological therapy, were dependent on alcohol or drugs, were actively suicidal or had attempted suicide in the previous 2 months, had psychosis or bipolar disorder, or were cognitively impaired were excluded. Intervention: CXA-UP or waitlist. Main Outcomes and Measures: Primary outcomes were changes in anxiety, depression, and somatic scores on the Patient Health Questionnaire and Posttraumatic Stress Disorder Checklist for DSM-5. Results: Among the 200 participants (160 women [80.0%]; 40 men [20.0%]; mean [SD] age, 43.1 [11.9] years), 120 were randomized to treatment and 80 to waitlist. Results for primary outcomes in the ITT analysis showed a significant pretreatment-to-posttreatment reduction when comparing treatment and waitlist on the posttraumatic stress disorder checklist for DSM-5 scores (slope [SE], -31.12 [3.00]; P < .001; Cohen d, 0.90; 90% CI, 0.63-1.19), 9-item Patient Health Questionnaire (PHQ-9) (slope [SE],-11.94 [1.30]; P < .001; Cohen d, 0.77; 90% CI, 0.52-1.06), PHQ-anxiety (slope [SE], -6.52 [0.67]; P < .001; Cohen d, 0.82; 90% CI, 0.49-1.15), and PHQ-somatic (slope [SE], -8.31 [0.92]; P < .001; Cohen d, 0.75; 90% CI, 0.47-1.04). Conclusions and Relevance: In this study, significant reductions and large effect sizes in all measures of different emotional disorders showed efficacy of a single transdiagnostic intervention in individuals exposed to armed conflicts. Trial Registration: ClinicalTrials.gov Identifier: NCT03127982.

Cancer-Associated Fibroblast-Like Tumor Cells Remodel the Ewing Sarcoma Tumor Microenvironment.

PURPOSE: Despite limited genetic and histologic heterogeneity, Ewing sarcoma (EwS) tumor cells are transcriptionally heterogeneous and display varying degrees of mesenchymal lineage specification in vitro. In this study, we investigated if and how transcriptional heterogeneity of EwS cells contributes to heterogeneity of tumor phenotypes in vivo. EXPERIMENTAL DESIGN: Single-cell proteogenomic-sequencing of EwS cell lines was performed and integrated with patient tumor transcriptomic data. Cell subpopulations were isolated by FACS for assessment of gene expression and phenotype. Digital spatial profiling and human whole transcriptome analysis interrogated transcriptomic heterogeneity in EwS xenografts. Tumor cell subpopulations and matrix protein deposition were evaluated in xenografts and patient tumors using multiplex immunofluorescence staining. RESULTS: We identified CD73 as a biomarker of highly mesenchymal EwS cell subpopulations in tumor models and patient biopsies. CD73+ tumor cells displayed distinct transcriptional and phenotypic properties, including selective upregulation of genes that are repressed by EWS::FLI1, and increased migratory potential. CD73+ cells were distinguished in vitro and in vivo by increased expression of matrisomal genes and abundant deposition of extracellular matrix (ECM) proteins. In epithelial-derived malignancies, ECM is largely deposited by cancer-associated fibroblasts (CAF), and we thus labeled CD73+ EwS cells, CAF-like tumor cells. Marked heterogeneity of CD73+ EwS cell frequency and distribution was detected in tumors in situ, and CAF-like tumor cells and associated ECM were observed in peri-necrotic regions and invasive foci. CONCLUSIONS: EwS tumor cells can adopt CAF-like properties, and these distinct cell subpopulations contribute to tumor heterogeneity by remodeling the tumor microenvironment. See related commentary by Kuo and Amatruda, p. 5002.

High-accuracy detection of supraspinatus fatty infiltration in shoulder MRI using convolutional neural network algorithms.

Background: The supraspinatus muscle fatty infiltration (SMFI) is a crucial MRI shoulder finding to determine the patient's prognosis. Clinicians have used the Goutallier classification to diagnose it. Deep learning algorithms have been demonstrated to have higher accuracy than traditional methods. Aim: To train convolutional neural network models to categorize the SMFI as a binary diagnosis based on Goutallier's classification using shoulder MRIs. Methods: A retrospective study was performed. MRI and medical records from patients with SMFI diagnosis from January 1st, 2019, to September 20th, 2020, were selected. 900 T2-weighted, Y-view shoulder MRIs were evaluated. The supraspinatus fossa was automatically cropped using segmentation masks. A balancing technique was implemented. Five binary classification classes were developed into two as follows, A: 0, 1 v/s 3, 4; B: 0, 1 v/s 2, 3, 4; C: 0, 1 v/s 2; D: 0, 1, 2, v/s 3, 4; E: 2 v/s 3, 4. The VGG-19, ResNet-50, and Inception-v3 architectures were trained as backbone classifiers. An average of three 10-fold cross-validation processes were developed to evaluate model performance. AU-ROC, sensitivity, and specificity with 95% confidence intervals were used. Results: Overall, 606 shoulders MRIs were analyzed. The Goutallier distribution was presented as follows: 0 = 403; 1 = 114; 2 = 51; 3 = 24; 4 = 14. Case A, VGG-19 model demonstrated an AU-ROC of 0.991 ± 0.003 (accuracy, 0.973 ± 0.006; sensitivity, 0.947 ± 0.039; specificity, 0.975 ± 0.006). B, VGG-19, 0.961 ± 0.013 (0.925 ± 0.010; 0.847 ± 0.041; 0.939 ± 0.011). C, VGG-19, 0.935 ± 0.022 (0.900 ± 0.015; 0.750 ± 0.078; 0.914 ± 0.014). D, VGG-19, 0.977 ± 0.007 (0.942 ± 0.012; 0.925 ± 0.056; 0.942 ± 0.013). E, VGG-19, 0.861 ± 0.050 (0.779 ± 0.054; 0.706 ± 0.088; 0.831 ± 0.061). Conclusion: Convolutional neural network models demonstrated high accuracy in MRIs SMFI diagnosis.

Application of untargeted volatile profiling in inflammatory bowel disease research.

Inflammatory bowel disease (IBD) diagnosis depends on criteria based on histological, endoscopic, radiological, and clinical results. These studies show drawbacks as being expensive, invasive, and time-consuming. In this work, an untargeted metabolomic strategy based on the monitoring of volatile compounds in serum by headspace gas chromatography-mass spectrometry is proposed as a complementary, fast, and efficient test for IBD patient diagnosis. To develop the method and build a chemometric model that allows the IBD diagnosis, serum samples including IBD patients and healthy volunteers were collected. Analyses were performed by incubating 400 µL of serum for 10 min at 90 °C. For data processing, an untargeted metabolomic strategy was used. A total of 96 features were detected, of which a total of 10 volatile compounds could be identified and confirmed by means of the analysis of real standards. The chemometric treatment consisted of a discriminant analysis of orthogonal partial least squares (OPLS-DA) obtaining a 100% of classification rate, since all the analyzed samples were correctly classified.

Macrophage phagocytosis of SARS-CoV-2-infected cells mediates potent plasmacytoid dendritic cell activation.

Early and strong interferon type I (IFN-I) responses are usually associated with mild COVID-19 disease, whereas persistent or unregulated proinflammatory cytokine responses are associated with severe disease outcomes. Previous work suggested that monocyte-derived macrophages (MDMs) are resistant and unresponsive to SARS-CoV-2 infection. Here, we demonstrate that upon phagocytosis of SARS-CoV-2-infected cells, MDMs are activated and secrete IL-6 and TNF. Importantly, activated MDMs in turn mediate strong activation of plasmacytoid dendritic cells (pDCs), leading to the secretion of high levels of IFN-α and TNF. Furthermore, pDC activation promoted IL-6 production by MDMs. This kind of pDC activation was dependent on direct integrin-mediated cell‒cell contacts and involved stimulation of the TLR7 and STING signaling pathways. Overall, the present study describes a novel and potent pathway of pDC activation that is linked to the macrophage-mediated clearance of infected cells. These findings suggest that a high infection rate by SARS-CoV-2 may lead to exaggerated cytokine responses, which may contribute to tissue damage and severe disease.

Use of polypyrrole ferrite microparticles and liquid chromatography-mass spectrometry for testing natural grass contamination by multiclass mycotoxins.

An analytical methodology based on the combination of dispersive magnetic solid-phase extraction (DMSPE) and liquid chromatography-mass spectrometry (LC-MS) is proposed to explore the occurrence of 13 mycotoxins (aflatoxins B1, G1, B2, and G2; deoxynivalenol; T-2 toxin; ochratoxin A; HT-2 toxin; enniatins A, A1, B, and B2; and beauvericin) and their derivatives in natural grass samples. Magnetic microparticles (Fe3O4) coated with polypyrrole (PPy) polymer were used in DMSPE sample treatment as adsorbent phase, and Fourier-transform infrared spectroscopy, field emission scanning electron microscopy, and energy dispersive X-ray spectroscopy have been used for its characterization. The experimental parameters influencing the adsorption and desorption steps of DMSPE have been optimized. Method validation has been carried out obtaining limits of quantification between 0.07 and 92 µg kg-1 corresponding to enniatin B or A1 and DON, respectively. A total of 83 natural grass samples from 8 dehesa farms were analysed. Enniatin B was found in all the samples (0.29 to 488 µg kg-1 concentration range) followed by enniatin B1 (92.8% of the samples) with a 0.12-137 µg kg-1 concentration range. Moreover, co-occurrence of mycotoxins was studied and between 2 and 5 mycotoxins appeared simultaneously in 97.6% of the samples. Distribution of the contamination according to natural grass location was also investigated.

Macroscopic analogue to entangled polymers.

The entangled structure of polymeric materials is often described as resembling a bowl of spaghetti, swarms of earthworms, or snakes. These analogies not only illustrate the concept, but form the foundation of polymer physics. However, the similarity between these macroscopic, athermal systems and polymers in terms of topology remains uncertain. To better understand this relationship, we conducted an experiment using X-ray tomography to study the structure of arrays of linear rubber bands. We found that, similar to linear polymers, the average number of entanglements increases linearly with the length of the ribbons. Additionally, we observed that entanglements are less frequent near the surface of the container, where there are also more ends, similar to what has been seen in trapped polymers. These findings provide the first experimental evidence supporting the visualization of polymer structures using macroscopic, athermal analogues, confirming the initial intuitive insights of the pioneers of polymer physics.

Spatial Distribution and Antioxidant Activity of Extracts from Citrus Fruits.

Citrus fruits are recommended components of the human diet because of their enriched composition in bioactive compounds and health benefits. Among their notable components are phenols, with a special emphasis on flavonoids, limonoids, and carboxylic acids. In this research, we have carried out a spatial metabolomics analysis for the characterization of these bioactive families in three citrus fruits, namely, lemons, limes, and mandarins. Sampling was undertaken, for which the juices and three fruit tissues, namely, albedo, flavedo, and segments, were analyzed. This characterization allowed for the determination of 49 bioactive compounds in all the samples. The composition of the different extracts was correlated with the antioxidant capacity measured by the DPPH radical scavenging activity and ß-carotene bleaching assays. Flavonoids, found in the albedo and flavedo at higher concentrations, were the main components responsible for DPPH radical scavenging activity. On the other hand, the combined action of flavonoids and limonoids contributed to explaining the antioxidant activity measured by the ß-carotene bleaching assay. Generally, the antioxidant capacity of juices was lower than that estimated for extracts from citrus tissues.

Carcinoma-associated fibroblast-like tumor cells remodel the Ewing sarcoma tumor microenvironment.

Tumor heterogeneity is a major driver of cancer progression. In epithelial-derived malignancies, carcinoma-associated fibroblasts (CAFs) contribute to tumor heterogeneity by depositing extracellular matrix (ECM) proteins that dynamically remodel the tumor microenvironment (TME). Ewing sarcomas (EwS) are histologically monomorphous, mesenchyme-derived tumors that are devoid of CAFs. Here we identify a previously uncharacterized subpopulation of transcriptionally distinct EwS tumor cells that deposit pro-tumorigenic ECM. Single cell analyses revealed that these CAF-like cells differ from bulk EwS cells by their upregulation of a matrisome-rich gene signature that is normally repressed by EWS::FLI1, the oncogenic fusion transcription factor that underlies EwS pathogenesis. Further, our studies showed that ECM-depositing tumor cells express the cell surface marker CD73, allowing for their isolation ex vivo and detection in situ. Spatial profiling of tumor xenografts and patient biopsies demonstrated that CD73 + EwS cells and tumor cell-derived ECM are prevalent along tumor borders and invasive fronts. Importantly, despite loss of EWS::FLI1-mediated gene repression, CD73 + EwS cells retain expression of EWS::FLI1 and the fusion-activated gene signature, as well as tumorigenic and proliferative capacities. Thus, EwS tumor cells can be reprogrammed to adopt CAF-like properties and these transcriptionally and phenotypically distinct cell subpopulations contribute to tumor heterogeneity by remodeling the TME.

Structural and Dynamical Properties of Elastin-Like Peptides near Their Lower Critical Solution Temperature.

Elastin-like peptides (ELPs) are artificially derived intrinsically disordered proteins (IDPs) mimicking the hydrophobic repeat unit in the protein elastin. ELPs are characterized by a lower critical solution temperature (LCST) in aqueous media. Here, we investigate the sequence GVG(VPGVG)3 over a wide range of temperatures (below, around, and above the LCST) and peptide concentrations employing all-atom molecular dynamics simulations, where we focus on the role of intra- and interpeptide interactions. We begin by investigating the structural properties of a single peptide that demonstrates a hydrophobic collapse with temperature, albeit moderate, because the sequence length is short. We observe a change in the interaction between two peptides from repulsive to attractive with temperature by evaluating the potential of mean force, indicating an LCST-like behavior. Next, we explore dynamical and structural properties of peptides in multichain systems. We report the formation of dynamical aggregates with coil-like conformation, in which valine central residues play an important role. Moreover, the lifetime of contacts between chains strongly depends on the temperature and can be described by a power-law decay that is consistent with the LCST-like behavior. Finally, the peptide translational and internal motion are slowed by an increase in the peptide concentration and temperature.