Publications by authors named "M Ángeles Esteban"

1,028 Publications

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Neutrophil subtypes shape HIV-specific CD8 T-cell responses after vaccinia virus infection.

NPJ Vaccines 2021 Apr 12;6(1):52. Epub 2021 Apr 12.

Institute for Research in Biomedicine, Università della Svizzera Italiana, Bellinzona, Switzerland.

Neutrophils are innate immune cells involved in the elimination of pathogens and can also induce adaptive immune responses. Nα and Nβ neutrophils have been described with distinct in vitro capacity to generate antigen-specific CD8 T-cell responses. However, how these cell types exert their role in vivo and how manipulation of Nβ/Nα ratio influences vaccine-mediated immune responses are not known. In this study, we find that these neutrophil subtypes show distinct migratory and motility patterns and different ability to interact with CD8 T cells in the spleen following vaccinia virus (VACV) infection. Moreover, after analysis of adhesion, inflammatory, and migration markers, we observe that Nβ neutrophils overexpress the α4β1 integrin compared to Nα. Finally, by inhibiting α4β1 integrin, we increase the Nβ/Nα ratio and enhance CD8 T-cell responses to HIV VACV-delivered antigens. These findings provide significant advancements in the comprehension of neutrophil-based control of adaptive immune system and their relevance in vaccine design.
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http://dx.doi.org/10.1038/s41541-021-00314-7DOI Listing
April 2021

SUMOylation modulates the stability and function of PI3K-p110β.

Cell Mol Life Sci 2021 Apr 8. Epub 2021 Apr 8.

Centro de Investigación en Medicina Molecular (CIMUS), CIMUS, P2L7, Universidade de Santiago de Compostela and Instituto de Investigaciones Sanitarias (IDIS), Avda Barcelona, 15706, Santiago de Compostela, Spain.

Class I PI3K are heterodimers composed of a p85 regulatory subunit and a p110 catalytic subunit involved in multiple cellular functions. Recently, the catalytic subunit p110β has emerged as a class I PI3K isoform playing a major role in tumorigenesis. Understanding its regulation is crucial for the control of the PI3K pathway in p110β-driven cancers. Here we sought to evaluate the putative regulation of p110β by SUMO. Our data show that p110β can be modified by SUMO1 and SUMO2 in vitro, in transfected cells and under completely endogenous conditions, supporting the physiological relevance of p110β SUMOylation. We identify lysine residue 952, located at the activation loop of p110β, as essential for SUMOylation. SUMOylation of p110β stabilizes the protein increasing its activation of AKT which promotes cell growth and oncogenic transformation. Finally, we show that the regulatory subunit p85β counteracts the conjugation of SUMO to p110β. In summary, our data reveal that SUMO is a novel p110β interacting partner with a positive effect on the activation of the PI3K pathway.
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http://dx.doi.org/10.1007/s00018-021-03826-6DOI Listing
April 2021

Emerging SARS-CoV-2 Variants and Impact in Global Vaccination Programs against SARS-CoV-2/COVID-19.

Vaccines (Basel) 2021 Mar 11;9(3). Epub 2021 Mar 11.

Centro Nacional de Biotecnología, Department of Molecular and Cellular Biology, Consejo Superior de Investigaciones Científicas (CNB-CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.

The emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) variants in different continents is causing a major concern in human global health. These variants have in common a higher transmissibility, becoming dominant within populations in a short time, and an accumulation of a high number of mutations in the spike (S) protein, especially within the amino terminal domain (NTD) and the receptor binding domain (RBD). These mutations have direct implications on virus infection rates through higher affinity of S RBD for the cellular angiotensin-converting enzyme-2 (ACE-2) receptor. There are also signs of enhanced virulence, re-infection frequency, and increased resistance to the action of monoclonal and polyclonal antibodies from convalescence sera and in vaccinated individuals in regions where the variants spread dominantly. In this review, we describe the different SARS-CoV-2 variants that have thus far been identified in various parts of the world with mutational changes and biological properties as well as their impact in medical countermeasures and human health.
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http://dx.doi.org/10.3390/vaccines9030243DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999234PMC
March 2021

Ultrasonography study of the skin wound healing process in gilthead seabream (Sparus aurata).

J Fish Dis 2021 Mar 24. Epub 2021 Mar 24.

Immunobiology for Aquaculture Group, Department of Cell Biology and Histology, Faculty of Biology, University of Murcia, Murcia, Spain.

This work aimed to carry out an in vivo study of the skin healing process in gilthead seabream (Sparus aurata) after being experimentally wounded. Firstly, the structure of normal skin was studied by real-time ultrasonography (Vevo Lab, VisualSonics) and light microscopy. Besides this, experimental wounds were made on the left flank of each fish with a circular biopsy punch (8 mm diameter) below the lateral line. The healing process was assessed on live fish at 0, 6, 11 and 23 days post-wounding using the real-time ultrasonography in B-mode and Power Doppler mode (Vevo 3100 FUJIFILM, VisualSonics). Through the ultrasonography images, both the skin structure and the evolution of the changes that wounds originated in the surrounding tissues were studied in vivo over time. Concomitantly, the pattern of neovascularization in the wounded area was followed during the healing process and it was demonstrated that, although the neovascularization started very early after the skin damage, it was increased in wounded areas from day 11 post-wounding onwards. The results obtained proved the utility and power of using ultrasounds in fish to evaluate in vivo complex biological processes in real time, which are difficult to study by other methodologies. The present data shed some light on the reparation of external injuries in aquatic vertebrates.
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http://dx.doi.org/10.1111/jfd.13370DOI Listing
March 2021

Global Profiling of the Lysine Crotonylome in Different Pluripotent States.

Genomics Proteomics Bioinformatics 2021 Mar 18. Epub 2021 Mar 18.

Jingjie PTM BioLab (Hangzhou) Co.Ltd, Hangzhou 310018, China. Electronic address:

Pluripotent stem cells (PSCs) can be expanded in vitro in different culture conditions, resulting in a spectrum of cell states with distinct properties. Understanding how PSCs transition from one state to another, ultimately leading to lineage-specific differentiation, is important for developmental biology and regenerative medicine. Although there is significant information regarding gene expression changes controlling these transitions, less is known about post-translational modifications of proteins. Protein crotonylation is a newly discovered post-translational modification where lysine residues are modified with a crotonyl group. Here, we employed affinity purification of crotonylated peptides and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to systematically profile protein crotonylation in mouse PSCs in different states including ground, metastable, and primed states, as well as metastable PSCs undergoing early pluripotency exit. We successfully identified 3628 high-confidence crotonylated sites in 1426 proteins. These crotonylated proteins are enriched for factors involved in functions/processes related to pluripotency such as RNA biogenesis, central carbon metabolism, and proteasome function. Moreover, we found that increasing the cellular levels of crotonyl-coenzyme A (crotonyl-CoA) through crotonic acid treatment promotes proteasome activity in metastable PSCs and delays their differentiation, consistent with previous observations showing that enhanced proteasome activity helps to sustain pluripotency. Our atlas of protein crotonylation will be valuable for further studies of pluripotency regulation and may also provide insights into the role of metabolism in other cell fate transitions.
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http://dx.doi.org/10.1016/j.gpb.2021.01.004DOI Listing
March 2021

[Retrospective study of lung carcinoid: experience in a third level Spanish hospital].

Rev Esp Patol 2021 Apr-Jun;54(2):85-91. Epub 2020 Oct 29.

Servicio de Oncología Médica, Hospital Universitario La Paz, Madrid, España.

Introduction: Pulmonary carcinoids are relatively rare neuroendocrine neoplasms, accounting for only 1-2% of malignant thoracic tumours. We describe our experience in the management and follow-up of such an infrequent tumour, with special emphasis on possible problems that might arise.

Patients And Methods: We present a descriptive retrospective study of all patients diagnosed with carcinoid tumour between January 2013 and January 2018. Demographic, histological and clinical data were collected and analyzed. Survival was recorded. SPSS version 21 was used for the statistical analysis.

Results: 42 patients with an average age of 66.26 years were included. The mean period of follow-up was 60 months and the average survival 59.12 months. The only statistically significant factor related to an improved survival time was tumour stage at diagnosis.

Conclusion: Carcinoid tumours are infrequent, which makes the objective collecting of data difficult. For this reason, we hope that the present study will contribute to a better understanding of their evolution.
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http://dx.doi.org/10.1016/j.patol.2020.04.004DOI Listing
October 2020

Incidence of COVID-19 in 902 Patients With Immunomediated Inflammatory Diseases Treated With Biologics and Targeted Synthetic Disease-Modifying Antirheumatic Drugs-Findings From the BIOCOVID Study.

J Clin Rheumatol 2021 Mar 5. Epub 2021 Mar 5.

From the Rheumatology Section, Hospital Universitario Infanta Sofía, Universidad Europea de Madrid Rheumatology Section, Hospital Universitario Infanta Leonor Faculty of Biomedical and Health Sciences, Universidad Europea de Madrid Pharmacy Section, Hospital Universitario Infanta Sofía, Universidad Europea de Madrid Rheumatology Section, Hospital Universitario Infanta Sofía Ophthalmology Section Gastroenterology Section, Hospital Universitario Infanta Sofía, Universidad Europea de Madrid Pharmacy Service Rheumatology Section, Hospital Universitario Infanta Leonor, Universidad Complutense de Madrid, Madrid, Spain.

Objectives: The aim of this study was to examine the incidence of coronavirus disease 2019 (COVID-19) among patients with immunomediated inflammatory diseases (IMIDs) treated with biologic or targeted synthetic disease-modifying antirheumatic drugs (bDMARDs and tsDMARDs) and to evaluate the influence of either IMIDs or related therapies on the incidence and evolution of COVID-19.

Methods: This observational, cross-sectional study was conducted from January 31, 2020, to May 15, 2020. Data of 902 patients were obtained from clinical records in hospitals, primary care units, and community pharmacies. Inclusion criteria were adults with IMIDs treated with bDMARDs or tsDMARDs who started therapy 3 months prior to study commencement. Patients with poor adherence to treatments were excluded. COVID-19 was classified as "definitive" (severe acute respiratory syndrome coronavirus 2 polymerase chain reaction [PCR]-positive), "possible" (characteristic symptoms and negative PCR), and "suspected" (characteristic symptoms but PCR not performed).

Results: COVID-19 was diagnosed in 70 patients (11 definitive, 19 possible, and 40 suspected). The cumulative incidence of definitive COVID-19 was 1.2%. When considering all cases, the incidence was 7.8%. Patients on biosimilars tumor necrosis factor blockers were more likely to have a diagnosis of COVID-19 (odds ratio, 2.308; p < 0.001). Patients on anti-B-cell therapies had a lower incidence of infections (p = 0.046). Low rates of hospitalization (14.3%), pneumonia (14.3%), death (2.9%), or thrombosis (2.9%) were observed, and 94.3% of patients recovered.

Conclusions: The cumulative incidence of confirmed cases of COVID-19 was similar to the general population, with generally low hospitalization, intensive care management, and mortality rates. COVID-19 incidence was less frequent in patients with more severe immunosuppression.
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http://dx.doi.org/10.1097/RHU.0000000000001716DOI Listing
March 2021

Daily rhythms in the morphometric parameters of hepatocytes and intestine of the European sea bass (Dicentrarchus labrax): influence of feeding time and hepatic zonation.

J Comp Physiol B 2021 May 23;191(3):503-515. Epub 2021 Feb 23.

Department of Physiology, Faculty of Biology, University of Murcia, 30100, Murcia, Spain.

The digestive system presents daily rhythms at both physiological and histological levels. Although cell morphology rhythms in mammals have been reported, they have scarcely been investigated in fish. The aim of the present research was to investigate the existence of daily rhythms in the morphology of cells in the liver and intestine of a teleost fish, the European sea bass (Dicentrarchus labrax), and how feeding time influences them. Regarding liver, we also focused on differences between the two metabolic zones: perivenous and periportal. For this purpose, fish were divided into two groups: fish fed once a day in the mid-light phase (ML) or the mid-dark phase (MD). After 1 month under each feeding regime, liver and intestine samples were collected every 4 h during a 24-h cycle, and different parameters were studied by light microscopy and image analysis. Daily rhythms occurred in most of the parameters evaluated in the liver. The effect of feeding time depended on the metabolic zone: the rhythms in the periportal zone were synchronized mainly by the light/dark cycle regardless of feeding time, whereas in the perivenous zone, rhythms were influenced more by feeding time. In the intestine, a daily rhythm in villi height was found with acrophases coinciding with feeding time in each group. These findings show for the first time the existence of cellular morphological rhythms in fish liver and intestine, and highlight the interactions between light and feeding cycles in the different metabolic zones of the liver.
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http://dx.doi.org/10.1007/s00360-020-01334-wDOI Listing
May 2021

Interference of anti-streptavidin antibodies: More common than we thought? In relation to six confirmed cases.

Clin Biochem 2021 Apr 2;90:62-65. Epub 2021 Feb 2.

Laboratorio Domecq & Lafage, Hospital Alemán, Buenos Aires, Argentina.

Automated immunoassays are extensively used in routine laboratory diagnostics of endocrine disorders because of their advantages, such as high sensitivity, precision, and specificity. However, these methods are limited by the susceptibility of the immunochemical reaction to various interferences. They may present interferences related to the assay's design, for example, the endogenous presence of anti-streptavidin antibodies (ASA) in platforms that use the biotin-streptavidin interaction. To date, there have been few reports in the literature of interference from endogenous ASA. However, such antibodies would potentially lead to falsely decreased or increased results of hormones that can lead to incorrect diagnoses. We report six patients with unusual thyroid function tests, incongruent to their clinical findings. They present elevated concentrations of total T3 and T4 and TSH values within the reference range when measured at Cobas 8000® e801 module (Roche Diagnostics®). Neither patient had been taking biotin; however, all demonstrated the presence of ASA causing falsely high results on competitive assays and also falsely low results on sandwich assays. The hormone panel was also analyzed in the same samples using a different platform available in our laboratory: Cobas 6000® e601 module (Roche Diagnostics®). Nine samples were sent to an external laboratory to be measured with the chemiluminescent method: ADVIA Centaur® (Siemens® Healthcare Diagnostics). The interference seems to affect e801 module and competitive assays the most without affecting results obtained by this chemiluminescent method. This interference could potentially affect other assays performed on the same platform, such as ATPO and estradiol. Finally, laboratories should suspect the presence of interference when there is no correlation between the hormone profile and the patient's clinic. The biotin neutralization protocol demonstrated its effectiveness to eliminate ASA interference.
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http://dx.doi.org/10.1016/j.clinbiochem.2021.01.013DOI Listing
April 2021

Changes in rainbow trout (Oncorhynchus mykiss) growth and mucosal immune parameters after dietary administration of grape (Vitis vinifera) seed extract.

Fish Physiol Biochem 2021 Apr 4;47(2):547-563. Epub 2021 Feb 4.

Department of Clinical Science, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, Iran.

The effect of dietary grape (Vitis vinifera) seed extract (GSE) on growth performance and mucosal immune parameters in rainbow trout (Oncorhynchus mykiss) fry was studied. Fish (1.3 g mean weight) were randomly distributed in nine tanks (15 fish per tank) and fed diets containing GSE at 0 (control), 100, and 200 mg kgfor 60 days. The results showed that growth parameters were enhanced in both treatment groups compared to the control group. Histological examination of fish skin showed higher epidermis thickness, goblet cell density, and volume density in the GSE groups compared to the values of the control group. Furthermore, the villus height, goblet cell density, and intraepithelial lymphocytes were increased in the fish intestine in those fish fed GSE, with respect to control fish. Feeding fish with low dose of GSE (100 mg kg) up-regulated the expression of some immune-relevant genes, including complement component 3 (C3), lysozyme (Lys), omDB-3, interferon gamma (IFN-γ), and tumor necrosis factor-α (TNF-α) in different mucosal tissues. However, feeding fish the high dose of GSE (200 mg kg) mostly enhanced expression of these genes in the skin. Besides, skin mucus of fish fed GSE showed bactericidal activity against Yersinia ruckeri. It was concluded that GSE, especially at 100 mg kg, modulates the growth performance and mucosal immunity of rainbow trout.
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http://dx.doi.org/10.1007/s10695-021-00930-zDOI Listing
April 2021

Impact of grape pomace flour (GPF) on immunity and immune-antioxidant-anti-inflammatory genes expression in Labeo rohita against Flavobacterium columnaris.

Fish Shellfish Immunol 2021 Apr 27;111:69-82. Epub 2021 Jan 27.

Department of Fish Biology and Ecology, Central Laboratory for Aquaculture Research, Abbassa, Abo-Hammad, Sharqia, Egypt.

This study evaluates the effects of dietary inclusion of grape pomace flour (GPF) on growth, antioxidant, anti-inflammatory, innate-adaptive immunity, and immune genes expression in Labeo rohita against Flavobacterium columnaris. In both normal and challenged fish the growth rate, hematology and biochemical parameters significantly increased when fed with 200 and 300 mg GPF enriched diets; similarly the activities of antioxidants and innate-adaptive immune parameters, such as malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH), phagocytic (PC), respiratory burst (RB), alternative pathway complement (ACP), lysozyme (Lyz), and total immunoglobulin M (IgM) significantly increased in both groups. Similarly, the immune, antioxidant, and anti-inflammatory-related gene mRNA expression was significantly up-regulated in head kidney (HK) tissues. The challenged fish fed without GPF always exhibited lower values of all the studied parameters. The results indicate that both normal and challenged fish treated with 200 mg GPF inclusion diet had significantly enhanced growth rate, antioxidant status, and immune defense mechanisms than with 300 mg GPF diet in L. rohita against F. columnaris.
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http://dx.doi.org/10.1016/j.fsi.2021.01.011DOI Listing
April 2021

COVID-19 vaccine candidates based on modified vaccinia virus Ankara expressing the SARS-CoV-2 spike induce robust T- and B-cell immune responses and full efficacy in mice.

J Virol 2021 Jan 7. Epub 2021 Jan 7.

Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología (CNB), Consejo Superior de Investigaciones Científicas (CSIC), 28049 Madrid, Spain;

Vaccines against SARS-CoV-2, the causative agent of the COVID-19 pandemic, are urgently needed. We developed two COVID-19 vaccines based on modified vaccinia virus Ankara (MVA) vectors expressing the entire SARS-CoV-2 spike (S) protein (MVA-CoV2-S); their immunogenicity was evaluated in mice using DNA/MVA or MVA/MVA prime/boost immunizations. Both vaccines induced robust, broad and polyfunctional S-specific CD4+ (mainly Th1) and CD8+ T-cell responses, with a T effector memory phenotype. DNA/MVA immunizations elicited higher T-cell responses. All vaccine regimens triggered high titers of IgG antibodies specific for the S, as well as for the receptor-binding domain; the predominance of the IgG2c isotype was indicative of Th1 immunity. Notably, serum samples from vaccinated mice neutralized SARS-CoV-2 in cell cultures, and those from MVA/MVA immunizations showed a higher neutralizing capacity. Remarkably, one or two doses of MVA-CoV2-S protect humanized K18-hACE2 mice from a lethal dose of SARS-CoV-2. In addition, two doses of MVA-CoV2-S confer full inhibition of virus replication in the lungs. These results demonstrate the robust immunogenicity and full efficacy of MVA-based COVID-19 vaccines in animal models and support its translation to the clinic. The continuous dissemination of the novel emerging SARS-CoV-2 virus, with more than 78 million infected cases worldwide and higher than 1,700,000 deaths as of December 23, 2020, highlights the urgent need for the development of novel vaccines against COVID-19. With this aim, we have developed novel vaccine candidates based on the poxvirus modified vaccinia virus Ankara (MVA) strain expressing the full-length SARS-CoV-2 spike (S) protein, and we have evaluated their immunogenicity in mice using DNA/MVA or MVA/MVA prime/boost immunization protocols. The results showed the induction of a potent S-specific T-cell response and high titers of neutralizing antibodies. Remarkably, humanized K18-hACE2 mice immunized with one or two doses of the MVA-based vaccine were 100% protected from SARS-CoV-2 lethality. Moreover, two doses of the vaccine prevented virus replication in lungs. Our findings prove the robust immunogenicity and efficacy of MVA-based COVID-19 vaccines in animal models and support its translation to the clinic.
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http://dx.doi.org/10.1128/JVI.02260-20DOI Listing
January 2021

TDP-43 aggregation induced by oxidative stress causes global mitochondrial imbalance in ALS.

Nat Struct Mol Biol 2021 02 4;28(2):132-142. Epub 2021 Jan 4.

Department of Cellular and Molecular Medicine, Institute of Genomic Medicine, University of California, San Diego, USA.

Amyotrophic lateral sclerosis (ALS) was initially thought to be associated with oxidative stress when it was first linked to mutant superoxide dismutase 1 (SOD1). The subsequent discovery of ALS-linked genes functioning in RNA processing and proteostasis raised the question of how different biological pathways converge to cause the disease. Both familial and sporadic ALS are characterized by the aggregation of the essential DNA- and RNA-binding protein TDP-43, suggesting a central role in ALS etiology. Here we report that TDP-43 aggregation in neuronal cells of mouse and human origin causes sensitivity to oxidative stress. Aggregated TDP-43 sequesters specific microRNAs (miRNAs) and proteins, leading to increased levels of some proteins while functionally depleting others. Many of those functionally perturbed gene products are nuclear-genome-encoded mitochondrial proteins, and their dysregulation causes a global mitochondrial imbalance that augments oxidative stress. We propose that this stress-aggregation cycle may underlie ALS onset and progression.
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http://dx.doi.org/10.1038/s41594-020-00537-7DOI Listing
February 2021

Immunity and inflammatory responses in gilthead sea bream (Sparus aurata L.) exposed to sub-lethal mixture of carbamazepine, cadmium chloride and polybrominated diphenyl ether.

Fish Shellfish Immunol 2021 Apr 23;111:25-35. Epub 2020 Dec 23.

University of Palermo, Dept. of Earth and Marine Science DISTEM, Laboratory of Marine Biochemistry and Ecotoxicology, Via Barlotta 4, 91100, Trapani, Italy; Istituto per lo studio degli impatti Antropici e Sostenibilità in ambiente marino (IAS), Consiglio Nazionale delle Ricerche, Capo Granitola, Trapani, Italy; Consorzio Universitario della Provincia di Trapani, Marine Biology Institute, Via Barlotta 4, 91100, Trapani, Italy.

Chemical contaminants such as industrial and urban by-products, pharmaceuticals, drugs metabolites and, plastics, are continuously found in the oceans, affecting its quality and organism's welfare. Although these compounds are found at concentrations ranged ng L, there is an increasing concern about the potential adverse effects of the interactions among those substances present, simultaneously, in a mixture. In the present study, specimens of sea bream (Sparus aurata) were exposed, by food, to rising concentrations of a mixture of carbamazepine, polybrominated diphenyl ether-47 and cadmium chloride, for 15 days and then, maintained, with the same control diet, without contaminants, for other 15 days. Samples of skin mucus, serum, head-kidney, liver and intestine were sampled at 0, 15 and 30 days. Cellular immune parameters were evaluated on head-kidney, as well as humoral parameters were determined on skin mucus and serum. In addition, the expression of some genes, related to immunity, was analysed on liver and intestine. Both cellular and humoral response were affected at 15 days, showing slightly signs of recovery at 30 days. Besides, the expression of immune-related genes was highly affected, suggesting the development of inflammatory processes, as well as a reduction of immune parameters. Overall, the mixture of compounds severally affected the immune system of sea bream, suggesting a lower degree of recovery. The prolonged exposure to a mixture of these compounds could entail serious change on population immunity and, eventually, promote changes on marine biota.
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http://dx.doi.org/10.1016/j.fsi.2020.12.013DOI Listing
April 2021

Single-cell landscape of the ecosystem in early-relapse hepatocellular carcinoma.

Cell 2021 Jan 23;184(2):404-421.e16. Epub 2020 Dec 23.

Department of Liver Surgery & Transplantation, Liver Cancer Institute, Zhongshan Hospital, Fudan University, and Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, China; Zhong-Hua Precision Medical Center, Zhongshan Hospital, Fudan University-BGI, Shanghai 200032, China. Electronic address:

Hepatocellular carcinoma (HCC) has high relapse and low 5-year survival rates. Single-cell profiling in relapsed HCC may aid in the design of effective anticancer therapies, including immunotherapies. We profiled the transcriptomes of ∼17,000 cells from 18 primary or early-relapse HCC cases. Early-relapse tumors have reduced levels of regulatory T cells, increased dendritic cells (DCs), and increased infiltrated CD8 T cells, compared with primary tumors, in two independent cohorts. Remarkably, CD8 T cells in recurrent tumors overexpressed KLRB1 (CD161) and displayed an innate-like low cytotoxic state, with low clonal expansion, unlike the classical exhausted state observed in primary HCC. The enrichment of these cells was associated with a worse prognosis. Differential gene expression and interaction analyses revealed potential immune evasion mechanisms in recurrent tumor cells that dampen DC antigen presentation and recruit innate-like CD8 T cells. Our comprehensive picture of the HCC ecosystem provides deeper insights into immune evasion mechanisms associated with tumor relapse.
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http://dx.doi.org/10.1016/j.cell.2020.11.041DOI Listing
January 2021

Molecular characterization of the T cell costimulatory receptors CD28 and CTLA4 in the European sea bass.

Fish Shellfish Immunol 2021 Feb 18;109:106-115. Epub 2020 Dec 18.

Immunobiotechnology for Aquaculture Group, Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100, Murcia, Spain. Electronic address:

For the activation of T cells, it is necessary the specific recognition of the peptide by the T cell receptors (TCR) in the surface of antigen-presenting cells (APCs) and additional signals delivered by costimulatory receptors. In fish, knowledge about the presence of these costimulatory signals is limited and functional evidence almost absent. Thus, in this study, we have identified the stimulatory CD28 and the inhibitory cytotoxic T-lymphocyte-associated protein 4 (CTLA4) coreceptors in the European sea bass (Dicentrarchus labrax), and evaluated their transcription. In parallel, the transcription encoding for the T cell markers CD8α and CD4 was also evaluated. Both coreceptors showed the canonical architecture including a signal peptide, an immunoglobulin domain, a transmembrane region and a cytosolic tail. Protein predictions and phylogenetic tree identify them as true mammalian orthologues of CD28 and CTLA4. We found these genes constitutively expressed in all studied organs of European sea bass with high expression in lymphoid organs (thymus, spleen and head-kidney) and liver. The molecular expression pattern of these genes was up-regulated in head-kidney leucocytes stimulated with T mitogens as concanavalin A and phytohemagglutinin (PHA), but not with the B cell mitogen lipopolysaccharide (LPS). Fish challenged with nodavirus (NNV) evidenced a differential and opposing regulation of the cd28 and ctla4 transcription levels in the brain, the target organ for viral replication, and head-kidney. While cd28 transcription tends to decrease over the infection time in both organs the expression of the ctla4 gene tends to increase. Interestingly, the coreceptor expression is highly and significantly correlated to the transcription of the T cell markers. Our results highlight the important role of CD28 and CTLA4 as costimulatory receptors of T cells in European sea bass but further studies are deserved.
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http://dx.doi.org/10.1016/j.fsi.2020.12.006DOI Listing
February 2021

Probiotic (SpPdp11) as a Fish Health Modulator: A Review.

Microorganisms 2020 Dec 14;8(12). Epub 2020 Dec 14.

Immunobiology for Aquaculture Group, Department of Cell Biology and Histology, Faculty of Biology, Campus Regional de Excelencia Internacional "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain.

Aquaculture is considered one of the largest food production sectors in the world. Probiotics have long been considered as a beneficial tool in this industry since these microorganisms improve the welfare of different fish species by modulating several physiological functions, such as metabolism, digestion, immune response, stress tolerance, and disease resistance, among others. SpPdp11, a probiotic isolated from the skin of healthy gilthead seabream, has been the center of attention in a good number of studies since its discovery. The purpose of this paper is to summarize, comment, and discuss the current knowledge related to the effects of SpPdp11 in two commercially important fish species in aquaculture (gilthead seabream and Senegalese sole). Furthermore, some considerations for future studies are also indicated.
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http://dx.doi.org/10.3390/microorganisms8121990DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7764857PMC
December 2020

Endoscopy-Related Bleeding and Thromboembolic Events in Patients on Direct Oral Anticoagulants or Vitamin K Antagonists.

Clin Gastroenterol Hepatol 2020 Dec 4. Epub 2020 Dec 4.

Department of Gastroenterology and Hepatology, Hospital Universitario Ramón y Cajal, Instituto Ramón y Cajal de Investigación Sanitaria, Centro de Investigación Biomédica en Red en el Área temática de Enfermedades Hepáticas y Digestivas, Universidad de Alcalá, Madrid, Spain.

Background & Aims: Few prospective studies have assessed the safety of direct oral anticoagulants (DOACs) in elective endoscopy. Our primary aim was to compare the risks of endoscopy-related gastrointestinal bleeding and thromboembolic events in patients on DOACs or vitamin K antagonists (VKAs) in this setting. Secondarily, we examined the impact of the timing of anticoagulant resumption on the risk of delayed bleeding in high-risk therapeutic procedures.

Methods: We conducted a multicenter, prospective, observational study from January 2018 to March 2020 of 1602 patients on oral anticoagulants (1004 on VKAs and 598 on DOACs) undergoing 1874 elective endoscopic procedures. Our primary outcomes were 90-day thromboembolic events and 30-day endoscopy-related gastrointestinal bleeding. The inverse probability of treatment weighting propensity score method was used for baseline covariate adjustment.

Results: The 2 groups had similar risks of endoscopy-related gastrointestinal bleeding (VKAs vs DOACs, 6.2% vs 6.7%; adjusted odds ratio [OR], 1.05; 95% CI, 0.67-1.65) and thromboembolic events (VKAs vs DOACs, 1.3% vs 1.5%; adjusted OR, 0.90; 95% CI, 0.34-2.38). In high bleeding risk procedures (n = 747), delayed anticoagulant resumption (> 48 hours or 24-48 hours vs < 24 hours) did not reduce the risk of postprocedural bleeding (10.3%, 9%, and 5.8%, respectively; adjusted P = .43). Hot and cold snare polypectomy were the most frequent high-risk interventions (41.8% and 39.8%, respectively).

Conclusion: In a prospective study of patients on DOACs or VKAs undergoing elective endoscopy, endoscopy-related bleeding and thromboembolic events showed similar risk. Our study suggests that early anticoagulant resumption is safe in most patients, but more data are needed for advanced high-risk therapeutic procedures.
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http://dx.doi.org/10.1016/j.cgh.2020.11.037DOI Listing
December 2020

Immune Profiles Identification by Vaccinomics After MVA Immunization in Randomized Clinical Study.

Front Immunol 2020 10;11:586124. Epub 2020 Nov 10.

Sorbonne Université, Inserm, Centre d'Immunologie et des Maladies Infectieuses (CIMIParis), Paris, France.

Background: Our previous work has demonstrated the benefits of transcutaneous immunization in targeting Langerhans cells and preferentially inducing CD8 T-cell responses.

Methods: In this randomized phase Ib clinical trial including 20 HIV uninfected volunteers, we compared the safety and immunogenicity of the MVA recombinant vaccine expressing HIV-B antigen (MVA-B) by transcutaneous and intramuscular routes. We hypothesized that the quality of innate and adaptive immunity differs according to the route of immunization and explored the quality of the vector vaccine-induced immune responses. We also investigated the early blood transcriptome and serum cytokine levels to identify innate events correlated with the strength and quality of adaptive immunity.

Results: We demonstrate that MVA-B vaccine is safe by both routes, but that the quality and intensity of both innate and adaptive immunity differ significantly. Transcutaneous vaccination promoted CD8 responses in the absence of antibodies and slightly affected gene expression, involving mainly genes associated with metabolic pathways. Intramuscular vaccination, on the other hand, drove robust changes in the expression of genes involved in IL-6 and interferon signalling pathways, mainly those associated with humoral responses, and also some levels of CD8 response.

Conclusion: Thus, vaccine delivery route perturbs early innate responses that shape the quality of adaptive immunity.

Clinical Trial Registration: http://ClinicalTrials.gov, identifier PER-073-13.
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http://dx.doi.org/10.3389/fimmu.2020.586124DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7683801PMC
November 2020

Review of inflammation in fish and value of the zebrafish model.

J Fish Dis 2021 Feb 25;44(2):123-139. Epub 2020 Nov 25.

Department of Cell Biology and Histology, Faculty of Biology, Immunobiology for Aquaculture Group, University of Murcia, Murcia, Spain.

Inflammation is a crucial step in the development of chronic diseases in humans. Understanding the inflammation environment and its intrinsic mechanisms when it is produced by harmful stimuli may be a key element in the development of human disease diagnosis. In recent decades, zebrafish (Danio rerio) have been widely used in research, due to their exceptional characteristics, as a model of various human diseases. Interestingly, the mediators released during the inflammatory response of both the immune system and nervous system, after its integration in the hypothalamus, could also facilitate the detection of injury through the register of behavioural changes in the fish. Although there are many studies that give well-defined information separately on such elements as the recruitment of cells, the release of pro- and anti-inflammatory mediators or the type of neurotransmitters released against different triggers, to the best of our knowledge there are no reviews that put all this knowledge together. In the present review, the main available information on inflammation in zebrafish is presented in order to facilitate knowledge about this important process of innate immunity, as well as the stress responses and behavioural changes derived from it.
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http://dx.doi.org/10.1111/jfd.13310DOI Listing
February 2021

The alleviation of skin wound-induced intestinal barrier dysfunction via modulation of TLR signalling using arginine in gilthead seabream (Sparus aurata L).

Fish Shellfish Immunol 2020 Dec 18;107(Pt B):519-528. Epub 2020 Nov 18.

Immunobiology for Aquaculture, Department of Cell Biology and Histology, Faculty of Biology, Campus of International Excellence, Campus Mare Nostrum, University of Murcia, Murcia, Spain. Electronic address:

The present study sought to investigate the effect of arginine on the involvement of toll-like receptors (TLRs) in skin wound-induced intestinal barrier dysfunction in gilthead seabream (Sparus aurata L.). Two replicates of fish (n = 8) were fed a commercial diet (CON, total 2.75% arginine), CON diet enriched with 1% arginine (ARG1, total 3.65% arginine) and 2% arginine (ARG2, total 4.53% arginine) for 30 days. Half of the fish were sampled, whereas the others were injured and sampled 7 days post-wounding. The intestinal histology results showed that a more intense infiltration of mixed leucocytes was evident in the wounded fish, which was remarkably reduced in fish that were fed the ARG1 diet. Serum IgM levels were significantly higher in the ARG1 group than levels in the CON group at 7 days post-wounding. Compared with the fish in the CON group after wounding, dietary administration of 1% arginine markedly downregulated the gene expression of TLRs (TLR2 and TLR5), MyD88, and proinflammatory cytokines (CSF1R, IL-1β, and TNFα), but significantly enhanced the gene expression of IκBα, the anti-inflammatory cytokine TGF-β1, and tight junction proteins (tricellulin and occludin) in wounded fish. Furthermore, the ARG2 diet demonstrated no additional benefits on intestinal cells, compared to both the ARG1 and the CON diets, and it even appeared to induce negative effects. In summary, dietary administration of 1% arginine significantly inhibited intestinal inflammatory response and tight junction disruption in skin-wounded gilthead seabream by modulating TLR signalling in the intestine.
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http://dx.doi.org/10.1016/j.fsi.2020.11.017DOI Listing
December 2020

Enhancement of HIV-1 Env-Specific CD8 T Cell Responses Using Interferon-Stimulated Gene 15 as an Immune Adjuvant.

J Virol 2020 12 22;95(2). Epub 2020 Dec 22.

Department of Preventive Medicine and Public Health and Microbiology, Universidad Autónoma de Madrid, Madrid, Spain

Induction of the endogenous innate immune system by interferon (IFN) triggers the expression of many proteins that serve like alarm bells in the body, activating an immune response. After a viral infection, one of the genes activated by IFN induction is the IFN-stimulated gene 15 (), which encodes a ubiquitin-like protein that undergoes a reversible posttranslational modification (ISGylation). ISG15 protein can also act unconjugated, intracellularly and secreted, acting as a cytokine. Although ISG15 has an essential role in host defense responses to microbial infection, its role as an immunomodulator in the vaccine field remains to be defined. In this investigation, we showed that ISG15 exerts an immunomodulatory role in human immunodeficiency virus (HIV) vaccines. In mice, after priming with a DNA-ISG15 vector mixed with a DNA expressing HIV-1 gp120 (DNA-gp120), followed by a booster with a modified vaccinia virus Ankara (MVA) vector expressing HIV-1 antigens, both wild-type ISG15-conjugated (ISG15-wt) and mutant unconjugated (ISG15-mut) proteins act as immune adjuvants by increasing the magnitude and quality of HIV-1-specific CD8 T cells, with ISG15-wt providing better immunostimulatory activity than ISG15-mut. The HIV-1 Env-specific CD8 T cell responses showed a predominant T effector memory (TEM) phenotype in all groups. Moreover, the amount of DNA-gp120 used to immunize mice could be reduced 5-fold after mixing with DNA-ISG15 without affecting the potency and the quality of the HIV-1 Env-specific immune responses. Our study clearly highlights the potential use of the IFN-induced ISG15 protein as immune adjuvant to enhance immune responses to HIV antigens, suggesting that this molecule might be exploitable for prophylactic and therapeutic vaccine approaches against pathogens. Our study described the potential role of ISG15 as an immunomodulatory molecule in the optimization of HIV/AIDS vaccine candidates. Using a DNA prime-MVA boost immunization protocol, our results indicated an increase in the potency and the quality of the HIV-1 Env-specific CD8 T cell response. These results highlight the adjuvant potency of ISG15 to elicit improved viral antigen presentation to the immune system, resulting in an enhanced HIV-1 vaccine immune response. The DNA-ISG15 vector could find applicability in the vaccine field in combination with other nucleic acid-based vector vaccines.
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http://dx.doi.org/10.1128/JVI.01155-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944454PMC
December 2020

AEU positioning statement on transdermal drug administration: Determinant evolution of functional urologic therapy.

Actas Urol Esp 2020 11 13;44(9):571-573. Epub 2020 Oct 13.

Servicio de Urología. Hospital Universitario Clínico San Carlos, Madrid, España.

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http://dx.doi.org/10.1016/j.acuro.2020.09.001DOI Listing
November 2020

Carbon Footprint of a Port Infrastructure from a Life Cycle Approach.

Int J Environ Res Public Health 2020 10 12;17(20). Epub 2020 Oct 12.

Department of Civil Engineering, Universidad Europea de Madrid, 28670 Villaviciosa de Odón, Madrid, Spain.

One of the most important consequences caused by the constant development of human activity is the uncontrolled generation of greenhouse gases (GHG). The main gases (CO, CH, and NO) are illustrated by the carbon footprint. To determine the impact of port infrastructures, a Life Cycle Assessment approach is applied that considers construction and maintenance. A case study of a port infrastructure in Spain is analyzed. Main results reflect the continuous emission of GHG throughout the useful life of the infrastructure (25 years). Both machinery (85%) and materials (15%) are key elements influencing the obtained results (117,000 Tm CO2e).
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http://dx.doi.org/10.3390/ijerph17207414DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7599594PMC
October 2020

JMJD3 acts in tandem with KLF4 to facilitate reprogramming to pluripotency.

Nat Commun 2020 10 8;11(1):5061. Epub 2020 Oct 8.

Chinese Academy of Sciences (CAS) Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health (GIBH), CAS, 510530, Guangzhou, China.

The interplay between the Yamanaka factors (OCT4, SOX2, KLF4 and c-MYC) and transcriptional/epigenetic co-regulators in somatic cell reprogramming is incompletely understood. Here, we demonstrate that the histone H3 lysine 27 trimethylation (H3K27me3) demethylase JMJD3 plays conflicting roles in mouse reprogramming. On one side, JMJD3 induces the pro-senescence factor Ink4a and degrades the pluripotency regulator PHF20 in a reprogramming factor-independent manner. On the other side, JMJD3 is specifically recruited by KLF4 to reduce H3K27me3 at both enhancers and promoters of epithelial and pluripotency genes. JMJD3 also promotes enhancer-promoter looping through the cohesin loading factor NIPBL and ultimately transcriptional elongation. This competition of forces can be shifted towards improved reprogramming by using early passage fibroblasts or boosting JMJD3's catalytic activity with vitamin C. Our work, thus, establishes a multifaceted role for JMJD3, placing it as a key partner of KLF4 and a scaffold that assists chromatin interactions and activates gene transcription.
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http://dx.doi.org/10.1038/s41467-020-18900-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7545202PMC
October 2020

Telemedicine and smart working: Spanish adaptation of the European Association of Urology recommendations.

Actas Urol Esp 2020 12 11;44(10):644-652. Epub 2020 Sep 11.

Instituto de Cirugía Urológica Avanzada (ICUA), Madrid, España.

Introduction: Telemedicine provides remote clinical support through technology tools. It can facilitate medical care delivery while reducing unnecessary office visits. The COVID-19 outbreak has caused an abrupt change in our daily urological practice, where teleconsultations play a crucial role.

Objective: To provide practical recommendations for the effective use of technological tools in telemedicine.

Materials And Methods: A literature search was conducted on Medline until April 2020. We selected the most relevant articles related to «telemedicine» and «smart working» that could provide valuable information.

Results: Telemedicine refers to the use of electronic information and telecommunication tools to provide remote clinical health care support. Smart working is a working approach that uses new or existing technologies to improve performance. Telemedicine is becoming a useful and fundamental tool during the COVID-19 pandemic and will be even more in the future. It is time for us to officially give telemedicine the place it deserves in clinical practice, and it is our responsibility to adapt and familiarize with all the tools and possible strategies for its optimal implementation. We must guarantee that the quality of care received by patients and perceived by them and their families is of the highest standard.

Conclusions: Telemedicine facilitates remote specialized urological clinical support and solves problems caused by limited patient mobility or transfer, reduces unnecessary visits to clinics and is useful to reduce the risk of COVID-19 viral transmission.
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http://dx.doi.org/10.1016/j.acuro.2020.08.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7486047PMC
December 2020

Radiological characterization of gilthead seabream (Sparus aurata) by X-ray computed tomography.

J Fish Biol 2020 Nov 22;97(5):1440-1447. Epub 2020 Sep 22.

Fish Innate Immune System Group, Department of Cell Biology and Histology, Faculty of Biology, University of Murcia, Murcia, Spain.

In recent years, the increasing use of fish as new animal models in scientific research and the growth of fish farming (mainly for human consumption) have highlighted the need for advanced technology to deepen our knowledge of fish biology. Hence, the present study was carried out to radiologically analyse the whole body of gilthead seabream (Sparus aurata) specimens using X-ray computed tomography (CT). Images were acquired in an Albira SPECT/PET/CT tri-modal preclinical-scanner. Segmentation, measurements and three-dimensional reconstruction were made using the Carestream Molecular imaging Albira CT system in conjunction with Pmod, AMIDE and Amira software packages. The results showed that the density values of gilthead seabream are in the range -700 to +2500 HU for the whole body. We also determined the density ranges that topographically coincide with the swim bladder, soft tissues, fat, skin and skeleton. This work describes, validates and demonstrates the application of a fully automated image analysis technique to study and quantify fish body composition, whether segmented or as a whole. In addition, the basis for applying this image technique in other in vivo studies is established.
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http://dx.doi.org/10.1111/jfb.14510DOI Listing
November 2020

Electroanalysis from the past to the twenty-first century: challenges and perspectives.

J Solid State Electrochem 2020 Jun 21:1-9. Epub 2020 Jun 21.

Department of Chemical Engineering and Analytical Chemistry, University of Barcelona, Martí i Franquès 1-11, E08028 Barcelona, Spain.

A personal mini-review is presented on the history of electroanalysis and on their present achievements and future challenges. The manuscript is written from the subjective view of two generations of electroanalytical chemists that have witnessed for many years the evolution of this discipline.
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http://dx.doi.org/10.1007/s10008-020-04733-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7306008PMC
June 2020

β-Catenin safeguards the ground state of mousepluripotency by strengthening the robustness of the transcriptional apparatus.

Sci Adv 2020 Jul 17;6(29):eaba1593. Epub 2020 Jul 17.

Laboratory of Integrative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.

Mouse embryonic stem cells cultured with MEK (mitogen-activated protein kinase kinase) and GSK3 (glycogen synthase kinase 3) inhibitors (2i) more closely resemble the inner cell mass of preimplantation blastocysts than those cultured with SL [serum/leukemia inhibitory factor (LIF)]. The transcriptional mechanisms governing this pluripotent ground state are unresolved. Release of promoter-proximal paused RNA polymerase II (Pol2) is a multistep process necessary for pluripotency and cell cycle gene transcription in SL. We show that β-catenin, stabilized by GSK3 inhibition in medium with 2i, supplies transcriptional coregulators at pluripotency loci. This selectively strengthens pluripotency loci and renders them addicted to transcription initiation for productive gene body elongation in detriment to Pol2 pause release. By contrast, cell cycle genes are not bound by β-catenin, and proliferation/self-renewal remains tightly controlled by Pol2 pause release under 2i conditions. Our findings explain how pluripotency is reinforced in the ground state and also provide a general model for transcriptional resilience/adaptation upon network perturbation in other contexts.
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http://dx.doi.org/10.1126/sciadv.aba1593DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7439582PMC
July 2020

Absence of IgG antibodies among high-risk contacts of two confirmed cases of Crimean-Congo haemorrhagic fever in the autonomous region of Madrid (Spain).

J Infect Public Health 2020 Oct 20;13(10):1595-1598. Epub 2020 Aug 20.

Health Department of the Community of Madrid, Subdirectorate of Epidemiology, Madrid, Comunidad de Madrid, Spain.

Crimean-Congo haemorrhagic fever (CCHF) is a widely distributed tick-borne disease. In Spain, the disease has emerged as outbreak associated with high-risk exposures. Our goal was to evaluate the prevalence of antibodies against the CCHF virus (CCHFV) in high-risk contacts. A cross-sectional study was conducted. Three hundred eighty-six high-risk contacts were identified comprising family contacts and hospital workers who had attended the cases. Fifty-seven cases with closer exposure were selected. However, forty-nine cases participated in the study. IgG antibodies were detected by immunoenzymatic techniques. All determinations tested negative for anti-CCHFV IgG antibodies. Most of the responders were women (73.5%), and belong to the intensive care department (53.1%). In relation to other possible sources of exposures, 18.4% travelled to countries with CCHF transmission risk. No CCHF positivity was recorded among selected high-risk contacts. This highlights the importance of standard precautions which might have protected healthcare workers and care providers from CCHF infection.
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http://dx.doi.org/10.1016/j.jiph.2020.07.016DOI Listing
October 2020