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[Introduction]: It is now clear that coronavirus disease 19 (COVID-19) severity is associated with a dysregulated immune response, but the relative contributions of different immune cells is still not fully understood. SARS CoV-2 infection triggers marked changes in NK cell populations, but there are contradictory reports as to whether these effector lymphocytes play a protective or pathogenic role in immunity to SARS-CoV-2., [Methods]: To address this question we have analysed differences in the phenotype and function of NK cells in SARS-CoV-2 infected individuals who developed either very mild, or life-threatening COVID-19 disease., [Results]: Although NK cells from patients with severe disease appeared more activated and the frequency of adaptive NK cells was increased, they were less potent mediators of ADCC than NK cells from patients with mild disease. Further analysis of peripheral blood NK cells in these patients revealed that a population of NK cells that had lost expression of the activating receptor NKG2D were a feature of patients with severe disease and this correlated with elevated levels of cell free NKG2D ligands, especially ULBP2 and ULBP3 in the plasma of critically ill patients. In vitro, culture in NKG2DL containing patient sera reduced the ADCC function of healthy donor NK cells and this could be blocked by NKG2DL-specific antibodies., [Discussion]: These observations of reduced NK function in severe disease are consistent with the hypothesis that defects in immune surveillance by NK cells permit higher levels of viral replication, rather than that aberrant NK cell function contributes to immune system dysregulation and immunopathogenicity.

Ebola virus (EBOV) is a single-strand RNA virus belonging to the Filoviridae family, which has been associated to most Ebola virus disease outbreaks to date, including the West African and the North Kivu epidemics between 2013 and 2022. This unprecedented health emergency prompted the search for effective medical countermeasures. Following up on the carbazole hit identified in our previous studies, we synthetized a new series of compounds, which demonstrated to prevent EBOV infection in cells by acting as virus entry inhibitors. The in vitro inhibitory activity was evaluated through the screening against surrogate models based on viral pseudotypes and further confirmed using replicative EBOV. Docking and molecular dynamics simulations joined to saturation transfer difference–nuclear magnetic resonance (STD–NMR) and mutagenesis experiments to elucidate the biological target of the most potent compounds. Finally, in vitro metabolic stability and in vivo pharmacokinetic studies were performed to confirm their therapeutic potential.

The constant appearance of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VoCs) has jeopardized the protective capacity of approved vaccines against coronavirus disease-19 (COVID-19). For this reason, the generation of new vaccine candidates adapted to the emerging VoCs is of special importance. Here, we developed an optimized COVID-19 vaccine candidate using the modified vaccinia virus Ankara (MVA) vector to express a full-length prefusion-stabilized SARS-CoV-2 spike (S) protein, containing 3 proline (3P) substitutions in the S protein derived from the beta (B.1.351) variant, termed MVA-S(3Pbeta). Preclinical evaluation of MVA-S(3Pbeta) in head-to-head comparison to the previously generated MVA-S(3P) vaccine candidate, expressing a full-length prefusion-stabilized Wuhan S protein (with also 3P substitutions), demonstrated that two intramuscular doses of both vaccine candidates fully protected transgenic K18-hACE2 mice from a lethal challenge with SARS-CoV-2 beta variant, reducing mRNA and infectious viral loads in the lungs and in bronchoalveolar lavages, decreasing lung histopathological lesions and levels of proinflammatory cytokines in the lungs. Vaccination also elicited high titers of anti-S Th1-biased IgGs and neutralizing antibodies against ancestral SARS-CoV-2 Wuhan strain and VoCs alpha, beta, gamma, delta, and omicron. In addition, similar systemic and local SARS-CoV-2 S-specific CD4+ and CD8+ T-cell immune responses were elicited by both vaccine candidates after a single intranasal immunization in C57BL/6 mice. These preclinical data support clinical evaluation of MVA-S(3Pbeta) and MVA-S(3P), to explore whether they can diversify and potentially increase recognition and protection of SARS-CoV-2 VoCs.

We generated an optimized COVID-19 vaccine candidate based on the modified vaccinia virus Ankara (MVA) vector expressing a full-length prefusion-stabilized SARS-CoV-2 spike (S) protein, termed MVA-CoV2-S(3P). The S(3P) protein was expressed at higher levels (2-fold) than the non-stabilized S in cells infected with the corresponding recombinant MVA viruses. One single dose of MVA-CoV2-S(3P) induced higher IgG and neutralizing antibody titers against parental SARS-CoV-2 and variants of concern than MVA-CoV2-S in wild-type C57BL/6 and in transgenic K18-hACE2 mice. In immunized C57BL/6 mice, two doses of MVA-CoV2-S or MVA-CoV2-S(3P) induced similar levels of SARS-CoV-2-specific B- and T-cell immune responses. Remarkably, a single administration of MVA-CoV2-S(3P) protected all K18-hACE2 mice from morbidity and mortality caused by SARS-CoV-2 infection, reducing SARS-CoV-2 viral loads, histopathological lesions, and levels of pro-inflammatory cytokines in the lungs. These results demonstrated that expression of a novel full-length prefusion-stabilized SARS-CoV-2 S protein by the MVA poxvirus vector enhanced immunogenicity and efficacy against SARS-CoV-2 in animal models, further supporting MVA-CoV2-S(3P) as an optimized vaccine candidate for clinical trials.

The coronavirus (CoV) disease 2019 (COVID-19) is expected to become endemic in the coming years, meaning that the worldwide society shall be prepared to routinely manage the highly contagious respiratory SARS-CoV-2. A rapid and early diagnosis of the SARS-CoV-2 infection is crucial for controlling the spread of the disease, interrupting the transmission chain, and providing timely medical attention to patients. We introduce an innovative nanophotonic biosensor for the quantitative detection of viral particles in less than 20 minutes total assay time. The nanosensor, based on the bimodal waveguide (BiMW) interferometric technology, has been functionalized with novel bioengineered nanobodies (Nb) targeting the SARS-CoV-2 receptor-binding domain (RBD). Our approach relies on the direct capture of the viral particles, and the optimized methodology allows the detection of the SARS-CoV-2 virus with outstanding sensitivity, below 200 TCID50 per mL, being able to provide accurate viral load determination within a broad dynamic range (102–106 TCID50 per mL). Both the nanobodies and the sensor nanotechnology can be produced at a large scale with highly-efficient cost-effective procedures, and they are being integrated into a user-friendly point-of-care device for multiplexed and decentralized operation. The implementation of this unique biosensor in primary care assistance, hospitals, pharmacies, or private laboratories could greatly aid in the relief and descongestion of the sanitary systems and the clinical and social management of COVID-19.

Current coronavirus disease-19 (COVID-19) vaccines are administered by the intramuscular route, but this vaccine administration failed to prevent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus infection in the upper respiratory tract, mainly due to the absence of virus-specific mucosal immune responses. It is hypothesized that intranasal (IN) vaccination could induce both mucosal and systemic immune responses that blocked SARS-CoV-2 transmission and COVID-19 progression. Here, we evaluated in mice IN administration of three modified vaccinia virus Ankara (MVA)-based vaccine candidates expressing the SARS-CoV-2 spike (S) protein, either the full-length native S or a prefusion-stabilized [S(3P)] protein; SARS-CoV-2-specific immune responses and efficacy were determined after a single IN vaccine application. Results showed that in C57BL/6 mice, MVA-based vaccine candidates elicited S-specific IgG and IgA antibodies in serum and bronchoalveolar lavages, respectively, and neutralizing antibodies against parental and SARS-CoV-2 variants of concern (VoC), with MVA-S(3P) being the most immunogenic vaccine candidate. IN vaccine administration also induced polyfunctional S-specific Th1-skewed CD4+ and cytotoxic CD8+ T-cell immune responses locally (in lungs and bronchoalveolar lymph nodes) or systemically (in spleen). Remarkably, a single IN vaccine dose protected susceptible K18-hACE2 transgenic mice from morbidity and mortality caused by SARS-CoV-2 infection, with MVA-S(3P) being the most effective candidate. Infectious SARS-CoV-2 viruses were undetectable in lungs and nasal washes, correlating with high titers of S-specific IgGs and neutralizing antibodies against parental SARS-CoV-2 and several VoC. Moreover, low histopathological lung lesions and low levels of pro-inflammatory cytokines in lungs and nasal washes were detected in vaccinated animals. These results demonstrated that a single IN inoculation of our MVA-based vaccine candidates induced pote

The immune response promoted by SARS-CoV-2 vaccination is relevant to develop novel vaccines and optimized prevention strategies. We analyzed the adaptive immunity in healthy donors (HD) and convalescent individuals (CD), before and after administering BNT162b2 vaccine. Our results revealed specific changes in CD4+ T cell reactivity profile in vaccinated HD and CD, with an increase in S1 and S2 positive individuals, proportionally higher for S2. On the contrary, NCAP reactivity observed in HD and CD patients was no longer detectable after vaccination. Despite the substantial antibody response in CD, MPro-derived peptides did not elicit CD4+ lymphocyte activation in our assay in either condition. HD presented an increment in anti-S and anti-RBD IgG after first dose vaccination, which increased after the second vaccination. Conversely, anti-S and anti-RBD IgG and IgA titers increased in already positive CD after first dose administration, remaining stable after second dose inoculation. Interestingly, we found a strong significant correlation between S1-induced CD4+ response and anti-S IgA pre-vaccination, which was lost after vaccine administration.

The emergence of SARS-CoV-2 variants that escape from immune neutralization are challenging vaccines and antibodies developed to stop the COVID-19 pandemic. Thus, it is important to establish therapeutics directed toward multiple or specific SARS-CoV-2 variants. The envelope spike (S) glycoprotein of SARS-CoV-2 is the key target of neutralizing antibodies (Abs). We selected a panel of nine nanobodies (Nbs) from dromedary camels immunized with the receptor-binding domain (RBD) of the S, and engineered Nb fusions as humanized heavy chain Abs (hcAbs). Nbs and derived hcAbs bound with subnanomolar or picomolar affinities to the S and its RBD, and S-binding cross-competition clustered them in two different groups. Most of the hcAbs hindered RBD binding to its human ACE2 (hACE2) receptor, blocked cell entry of viruses pseudotyped with the S protein and neutralized SARS-CoV-2 infection in cell cultures. Four potent neutralizing hcAbs prevented the progression to lethal SARS-CoV-2 infection in hACE2-transgenic mice, demonstrating their therapeutic potential. Cryo-electron microscopy identified Nb binding epitopes in and out the receptor binding motif (RBM), and showed different ways to prevent virus binding to its cell entry receptor. The Nb binding modes were consistent with its recognition of SARS-CoV-2 RBD variants; mono and bispecific hcAbs efficiently bound all variants of concern except omicron, which emphasized the immune escape capacity of this latest variant.

We generated an optimized COVID-19 vaccine candidate based on the modified vaccinia virus Ankara (MVA) vector expressing a full-length prefusion-stabilized SARS-CoV-2 spike (S) protein, termed MVA-CoV2-S(3P). The S(3P) protein was expressed at higher levels (2-fold) than the non-stabilized S in cells infected with the corresponding recombinant MVA viruses. One single dose of MVA-CoV2-S(3P) induced higher IgG and neutralizing antibody titers against parental SARS-CoV-2 and variants of concern than MVA-CoV2-S in wild-type C57BL/6 and in transgenic K18-hACE2 mice. In immunized C57BL/6 mice, two doses of MVA-CoV2-S or MVA-CoV2-S(3P) induced similar levels of SARS-CoV-2-specific B- and T-cell immune responses. Remarkably, a single administration of MVA-CoV2-S(3P) protected all K18-hACE2 mice from morbidity and mortality caused by SARS-CoV-2 infection, reducing SARS-CoV-2 viral loads, histopathological lesions, and levels of pro-inflammatory cytokines in the lungs. These results demonstrated that expression of a novel full-length prefusion-stabilized SARS-CoV-2 S protein by the MVA poxvirus vector enhanced immunogenicity and efficacy against SARS-CoV-2 in animal models, further supporting MVA-CoV2-S(3P) as an optimized vaccine candidate for clinical trials.

Two doses of the MVA-CoV2-S vaccine candidate expressing the SARS-CoV-2 spike (S) protein protected K18-hACE2 transgenic mice from a lethal dose of SARS-CoV-2. This vaccination regimen prevented virus replication in the lungs, reduced lung pathology, and diminished levels of pro-inflammatory cytokines. High titers of IgG antibodies against S and receptor-binding domain (RBD) proteins and of neutralizing antibodies were induced against parental virus and variants of concern, markers that correlated with protection. Similar SARS-CoV-2-specific antibody responses were observed at prechallenge and postchallenge in the two-dose regimen, while the single-dose treatment does not avoid vaccine breakthrough infection. All vaccinated animals survived infection and were also protected to SARS-CoV-2 reinfection. Furthermore, two MVA-CoV2-S doses induced long-term memory S-specific humoral and cellular immune responses in C57BL/6 mice, 6 months after immunization. The efficacy and immunological benefits of the MVA-CoV2-S vaccine candidate against COVID-19 supports its consideration for human clinical trials.

Currently, there is a need for reliable tests that allow identification of individuals that have been infected with SARS-CoV-2 even if the infection was asymptomatic. To date, the vast majority of the serological tests for SARS-CoV-2–specific Abs are based on serum detection of Abs to either the viral spike glycoprotein (the major target for neutralizing Abs) or the viral nucleocapsid protein that is known to be highly immunogenic in other coronaviruses. Conceivably, exposure of Ags released from infected cells could stimulate Ab responses that might correlate with tissue damage and, hence, they may have some value as a prognostic indicator. We addressed whether other nonstructural viral proteins, not incorporated into the infectious viral particle, specifically the viral cysteine-like protease, might also be potent immunogens. Using ELISA tests, coating several SARS-CoV-2 proteins produced in vitro, we describe that COVID-19 patients make high titer IgG, IgM, and IgA Ab responses to the Cys-like protease from SARS-CoV-2, also known as 3CLpro or Mpro, and it can be used to identify individuals with positive serology against the coronavirus. Higher Ab titers in these assays associated with more-severe disease, and no cross-reactive Abs against prior betacoronavirus were found. Remarkably, IgG Abs specific for Mpro and other SARS-CoV-2 Ags can also be detected in saliva. In conclusion, Mpro is a potent Ag in infected patients that can be used in serological tests, and its detection in saliva could be the basis for a rapid, noninvasive test for COVID-19 seropositivity.

Currently, there is a need for reliable tests that allow identification of individuals that have been infected with SARS-CoV-2 even if the infection was asymptomatic. To date, the vast majority of the serological tests for SARS-CoV-2 specific antibodies are based on serum detection of antibodies to either the viral spike glycoprotein (the major target for neutralising antibodies) or the viral nucleocapsid protein that are known to be highly immunogenic in other coronaviruses. Conceivably, exposure of antigens released from infected cells could stimulate antibody responses that might correlate with tissue damage and, hence, they may have some value as a prognostic indicator. We addressed whether other non-structural viral proteins, not incorporated into the infectious viral particle, specifically the viral cysteine-like protease, might also be potent immunogens. Using ELISA tests, coating several SARS-CoV-2 proteins produced in vitro, we describe that COVID-19 patients make high titre IgG, IgM and IgA antibody responses to the Cys-like protease from SARS-CoV-2, also known as 3CLpro or Mpro, and it can be used to identify individuals with positive serology against the coronavirus. Higher antibody titres in these assays associated with more severe disease and no cross-reactive antibodies against prior betacoronavirus were found. Remarkably, IgG antibodies specific for Mpro and other SARS-CoV-2 antigens can also be detected in saliva. In conclusion, Mpro is a potent antigen in infected patients that can be used in serological tests and its detection in saliva could be the basis for a rapid, non-invasive test for COVID-19 seropositivity.

In the present invention, we herein generally provide an in vitro method for detecting in at least one biological sample an antibody that binds to at least one epitope of the SARS-CoV-2 virus, comprising contacting said at least one biological sample with at least one isolated SARS-CoV-2 Mpro protein, or at least one fragment of said isolated SARS-CoV-2 Mpro protein comprising at least one epitope of the SARS-CoV-2 virus, and detecting the formation of an antigen-antibody complex between said virus protein or said fragment and an antibody present in said biological sample.

Many new aspects of COVID-19 disease, including different clinical manifestations, have been identified during the pandemic. The wide array of symptoms and variation in disease severity after SARS-CoV-2 infection might be related to heterogeneity in the immune responses of different patients. Here we describe a new method for a simple multi-antigen serological test that generates a full picture of seroconversion in a single reaction. The assay is based on the detection by flow cytometry of multiple immunoglobulin classes (isotypes) specific for four SARS-CoV-2 antigens: the Spike glycoprotein (one of the highly immunogenic proteins), its RBD fragment (the major target for neutralising antibodies), the nucleocapsid protein and the main cysteine-like protease. Until now, most diagnostic serological tests measured antibodies to only one antigen and some patients seemed to not make any antibody response. Our data reveal that while most patients respond against all the viral antigens tested, others show a marked bias to make antibodies against either proteins exposed on the viral particle or those released after cellular infection. Combining all the four antigens and using machine learning techniques, it was possible to clearly discriminate between patients and healthy controls with 100% confidence. Further, combination of antigens and different immunoglobulin isotypes in this multi-antigen assay improved the classification of patients with mild and severe disease. Introduction of this method will facilitate massive screenings of patients to evaluate their immune response. It could also support vaccination campaigns both to select non-immune individuals and to distinguish infected patients from vaccine responders.

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.

Here, we describe a new, simple, highly multiplexed serological test that generates a more complete picture of seroconversion than single antigen-based assays. Flow cytometry is used to detect multiple Ig isotypes binding to four SARS-CoV-2 antigens: the Spike glycoprotein, its RBD fragment (the main target for neutralizing antibodies), the nucleocapsid protein, and the main cysteine-like protease in a single reaction. Until now, most diagnostic serological tests measured antibodies to only one antigen and in some laboratory-confirmed patients no SARS-CoV-2-specific antibodies could be detected. Our data reveal that while most patients respond against all the viral antigens tested, others show a marked bias to make antibodies against either proteins exposed on the viral particle or those released after cellular infection. With this assay, it was possible to discriminate between patients and healthy controls with 100% confidence. Analysing the response of multiple Ig isotypes to the four antigens in combination may also help to establish a correlation with the severity degree of disease. A more detailed description of the immune responses of different patients to SARS-CoV-2 virus might provide insight into the wide array of clinical presentations of COVID-19.

SARS‐CoV‐2 infection causes an abrupt response by the host immune system, which is largely responsible for the outcome of COVID‐19. We investigated whether the specific immune responses in the peripheral blood of 276 patients associated to severity and progression of COVID‐19. At admission, dramatic lymphopenia of T, B and NK cells associated to severity. Conversely, the proportion of B cells, plasmablasts, circulating follicular helper T cells (cTfh) and CD56‐CD16+ NK‐cells increased. Regarding humoral immunity, levels of IgM, IgA and IgG were unaffected, but when degrees of severity were considered, IgG was lower in severe patients. Compared to healthy donors, complement C3 and C4 protein levels were higher in mild and moderate, but not in severe patients, while the activation peptide of C5 (C5a) increased from the admission in every patient, regardless their severity. Moreover, total IgG, the IgG1 and IgG3 isotypes and C4 decreased from day 0 to day 10 in patients who were hospitalized for more than two weeks, but not in patients who were discharged earlier. Our study provides important clues to understand the immune response observed in COVID‐19 patients, associating severity with an imbalanced humoral response and identifying new targets for therapeutic intervention.

BACKGROUND: Although the role of Th17 and regulatory T cells in the progression of atherosclerosis has been highlighted in recent years, their molecular mediators remain elusive. We aimed to evaluate the association between the CD69 receptor, a regulator of Th17/regulatory T cell immunity, and atherosclerosis development in animal models and in patients with subclinical disease. METHODS: Low-density lipoprotein receptor-deficient chimeric mice expressing or not expressing CD69 on either myeloid or lymphoid cells were subjected to a high fat diet. In vitro functional assays with human T cells were performed to decipher the mechanism of the observed phenotypes. Expression of CD69 and NR4A nuclear receptors was evaluated by reverse transcription-polymerase chain reaction in 305 male participants of the PESA study (Progression of Early Subclinical Atherosclerosis) with extensive (n=128) or focal (n=55) subclinical atherosclerosis and without disease (n=122). RESULTS: After a high fat diet, mice lacking CD69 on lymphoid cells developed large atheroma plaque along with an increased Th17/regulatory T cell ratio in blood. Oxidized low-density lipoprotein was shown to bind specifically and functionally to CD69 on human T lymphocytes, inhibiting the development of Th17 cells through the activation of NR4A nuclear receptors. Participants of the PESA study with evidence of subclinical atherosclerosis displayed a significant CD69 and NR4A1 mRNA downregulation in peripheral blood leukocytes compared with participants without disease. The expression of CD69 remained associated with the risk of subclinical atherosclerosis in an adjusted multivariable logistic regression model (odds ratio, 0.62; 95% CI, 0.40-0.94; P=0.006) after adjustment for traditional risk factors, the expression of NR4A1, the level of oxidized low-density lipoprotein, and the counts of different leucocyte subsets. CONCLUSIONS: CD69 depletion from the lymphoid compartment promotes a Th17/regulatory T cell imbala

Acute hemorrhagic conjunctivitis (AHC) is a painful, contagious eye disease, with millions of cases in the last decades. Coxsackievirus A24 (CV-A24) was not originally associated with human disease, but in 1970 a pathogenic "variant" (CV-A24v) emerged, which is now the main cause of AHC. Initially, this variant circulated only in Southeast Asia, but it later spread worldwide, accounting for numerous AHC outbreaks and two pandemics. While both CV-A24 variant and nonvariant strains still circulate in humans, only variant strains cause AHC for reasons that are yet unknown. Since receptors are important determinants of viral tropism, we set out to map the CV-A24 receptor repertoire and establish whether changes in receptor preference have led to the increased pathogenicity and rapid spread of CV-A24v. Here, we identify ICAM-1 as an essential receptor for both AHC-causing and non-AHC strains. We provide a high-resolution cryo-EM structure of a virus-ICAM-1 complex, which revealed critical ICAM-1-binding residues. These data could help identify a possible conserved mode of receptor engagement among ICAM-1-binding enteroviruses and rhinoviruses. Moreover, we identify a single capsid substitution that has been adopted by all pandemic CV-A24v strains and we reveal that this adaptation enhances the capacity of CV-A24v to bind sialic acid. Our data elucidate the CV-A24v receptor repertoire and point to a role of enhanced receptor engagement in the adaptation to the eye, possibly enabling pandemic spread.

Acute hemorrhagic conjunctivitis (AHC) is a painful, contagious eye disease, with millions of cases in the last decades. Coxsackievirus A24 (CV-A24) was not originally associated with human disease, but in 1970 a pathogenic "variant" (CV-A24v) emerged, which is now the main cause of AHC. Initially, this variant circulated only in Southeast Asia, but it later spread worldwide, accounting for numerous AHC outbreaks and two pandemics. While both CV-A24 variant and nonvariant strains still circulate in humans, only variant strains cause AHC for reasons that are yet unknown. Since receptors are important determinants of viral tropism, we set out to map the CV-A24 receptor repertoire and establish whether changes in receptor preference have led to the increased pathogenicity and rapid spread of CV-A24v. Here, we identify ICAM-1 as an essential receptor for both AHC-causing and non-AHC strains. We provide a high-resolution cryo-EM structure of a virus-ICAM-1 complex, which revealed critical ICAM-1-binding residues. These data could help identify a possible conserved mode of receptor engagement among ICAM-1-binding enteroviruses and rhinoviruses. Moreover, we identify a single capsid substitution that has been adopted by all pandemic CV-A24v strains and we reveal that this adaptation enhances the capacity of CV-A24v to bind sialic acid. Our data elucidate the CV-A24v receptor repertoire and point to a role of enhanced receptor engagement in the adaptation to the eye, possibly enabling pandemic spread.

Cell surface aminopeptidase N (APN) is a membrane-bound ectoenzyme that hydrolyzes proteins and peptides and regulates numerous cell functions. APN participates in tumor cell expansion and motility, and is a target for cancer therapies. Small drugs that bind to the APN active site inhibit catalysis and suppress tumor growth. APN is also a major cell entry receptor for coronavirus, which binds to a region distant from the active site. Three crystal structures that we determined of human and pig APN ectodomains defined the dynamic conformation of the protein. These structures offered snapshots of closed, intermediate and open APN, which represent distinct functional states. Coronavirus envelope proteins specifically recognized the open APN form, prevented ectodomain progression to the closed form and substrate hydrolysis. In addition, drugs that bind the active site inhibited both coronavirus binding to cell surface APN and infection; the drugs probably hindered APN transition to the virus-specific open form. We conclude that allosteric inhibition of APN functions occurs by ligand suppression of ectodomain motions necessary for catalysis and virus cell entry, as validated by locking APN with disulfides. Blocking APN dynamics can thus be a valuable approach to development of drugs that target this ectoenzyme.

The prediction of grape composition is becoming more important due to the need of reducing the current levels of alcohol and pH of the wines, a problem that is exacerbated by climate change. This work presents a 3-year study of the spatial variability of grape composition in a rainfed Tempranillo vineyard located in Rioja (Spain). It is based on the acquisition of multispectral imagery at véraison (start of the ripening process); and zoning based on NDVI, to assess its performance for zonal management. The results reveal a high spatial variability within the plot, with a stable pattern over the years, even with very different climate conditions. NDVI was a good predictor of vegetative growth variables. However, the prediction of grape composition was more complex. Unexpectedly, anthocyanins were found to be higher in the highest vigor zone, which is probably related to the effects of climate change. This unexpected relationship is particularly discussed in the article.

Background: Physical inactivity remains a public health problem worldwide. The purpose of this study was to determine compliance with physical activity (PA) recommendations developed by the World Health Organization (WHO) and to analyze the association with different cardiovascular risk factors (CVRF) and sociodemographic and lifestyle determinants in a sample of Spanish workers. Methods: A cross sectional analysis was carried out in a sample of 2.651 individuals (2.428 men and 133 women, aged 40-55 years old) from the Aragon Workers' Health Study (AWHS). Sample was divided into two groups depending on whether they met PA recommendations during the last year, including transportation, leisure and occupational PA. Sociodemographic, anthropometric, clinical, pharmacological and lifestyle data were collected during the years 2011-2014. Median and interquartile range for quantitative variables and frequency distribution for categorical variables were used. Crude and adjusted binary logistic regression models were carried out to study the likelihood of meeting with PA recommendations based on the number of CVRF (hypertension, dyslipidemia, diabetes, obesity and smoking) present. Results: 47.0% of workers complied with WHO recommendations for PA. Lower prevalence of compliance was found among those participants presenting some CVRF (39.7% in hypertensive, 38.7% in dyslipidemic and 32.7% in diabetic). Of the total study sample, 51.3% reported not performing PA or practice a light PA, 20.2% moderate and 28.5% vigorous or very vigorous PA. The likelihood of compliance with PA recommendations decreased with increasing the number of CVRF: 1 CVRF (OR: 0.62; 95%IC: 0.49-0.78), 2 CVRF (OR: 0.49; 95% IC: 0.38-0.62), 3 CVRF (OR: 0.34; 95% IC: 0.25-0.46), 4 CVRF (OR: 0.19; 95% IC: 0.11-0.32). Conclusions: Compliance with recommendations of PA in our population remains insufficient, moreover in those with increased cardiovascular risk., Fundamentos: La inactividad física continúa siendo un problema de salud pública a nivel mundial. El objetivo del presente estudio fue determinar la adherencia a las recomendaciones de actividad física (AF) establecidas por la Organización Mundial de la Salud (OMS) de una muestra de trabajadores españoles, así como analizar su asociación con diferentes factores de riesgo cardiovascular (FRCV), indicadores sociodemográficos y de estilo de vida. Métodos: Estudio tranversal con 2.651 participantes en la cohorte del Aragon Workers' Health Study (AWHS) (2.428 varones y 133 mujeres), con edades comprendidas entre los 40-55 años. Se dividió a la población en dos grupos en función de si habían cumplido o no durante el último año las recomendaciones sobre AF establecidas por la OMS, teniendo en cuenta las actividades relacionadas con la ocupación, ocio y recreación y desplazamientos. Se registraron variables sociodemográficas, antropométricas, clínicas, farmacológicas y de estilo de vida entre los años 2011-2014. Se calcularon la mediana y el intervalo intercuartílico para variables cuantitativas y la distribución de frecuencias para las categóricas. Se estimó un modelo de regresión logística binaria crudo y ajustado para analizar la probabilidad de cumplir las recomendaciones de AF en función del número de FRCV (hipertensión, dislipemia, diabetes, obesidad y tabaquismo). Resultados: El 47,0% de participantes cumplían las recomendaciones. El cumplimiento fue del 39,7% en hipertensos, del 38,7% en dislipémicos y 32,7% en diabéticos. El 51,3% reportó no realizar AF o realizar una AF ligera, el 20,2% moderada y el 28,5% vigorosa o muy vigorosa. Los resultados mostraron cómo a medida que aumentaba el número de FRCV disminuía la probabilidad de adherirse a las recomendaciones: 1 FRCV (OR: 0,62; 95%CI: 0,49-0,78), 2 FRCV (OR: 0,49; 95%CI: 0,38-0,62), 3 FRCV (OR: 0,34; 95%CI: 0,25-0,46), 4 FRCV (OR: 0,19; 95%CI: 0,11-0,32). Conclusiones: El incumplimiento de las recomendaciones de A

The prediction of grape composition is becoming more important due to the need of reducing the current levels of alcohol and pH of the wines, a problem that is exacerbated by climate change. This work presents a 3-year study of the spatial variability of grape composition in a rainfed Tempranillo vineyard located in Rioja (Spain). It is based on the acquisition of multispectral imagery at véraison (start of the ripening process); and zoning based on NDVI, to assess its performance for zonal management. The results reveal a high spatial variability within the plot, with a stable pattern over the years, even with very different climate conditions. NDVI was a good predictor of vegetative growth variables. However, the prediction of grape composition was more complex. Unexpectedly, anthocyanins were found to be higher in the highest vigor zone, which is probably related to the effects of climate change. This unexpected relationship is particularly discussed in the article.

Últimamente ha cobrado gran interés el estudio de los compuestos de la uva que pueden desajustar su composición y la de los vinos con ella elaborados; los que determinan el grado alcohólico y el color, fundamentalmente. Ese desajuste se ve incrementado por las condiciones que impone el cambio climático. El presente trabajo contempla un estudio de 4 años de duración, realizado en un viñedo de secano de ‘Tempranillo’ localizado en la D.O.Ca. Rioja. Su objetivo principal fue evaluar las relaciones entre vigor de la planta, respuesta vegetativo-productiva y composición de la uva. Para ello, se tomaron imágenes aéreas multiespectrales en envero y se zonificó el viñedo en dos niveles diferentes de vigor basándose en el índice NDVI. Inesperadamente, la relación entre antocianos y vigor fue positiva en tres de los cuatro años de estudio, lo cual se discute particularmente en el artículo.

In the coronavirus (CoV), the envelope spike (S) glycoprotein is responsible for CoV cell entry and host-to-host transmission. The S is a multifunctional glycoprotein that mediates both attachment of CoV particles to cell surface receptor molecules as well as membrane penetration by fusion. Receptor-binding domains (RBD) have been identified in the S of diverse CoV; they usually contain antigenic determinants targeted by antibodies that neutralize CoV infections. To penetrate host cells, the CoV can use various cell surface molecules, although they preferentially bind to ectoenzymes. Several crystal structures have determined the folding of CoV RBD and the mode by which they recognize cell entry receptors. Here we review the CoV-receptor complex structures reported to date, and highlight the distinct receptor recognition modes, common features, and key determinants of the binding specificity. Structural studies have established the basis for understanding receptor recognition diversity in CoV, its evolution and the adaptation of this virus family to different hosts. CoV responsible for recent outbreaks have extraordinary potential for cross-species transmission; their RBD bear large platforms specialized in recognition of receptors from different species, which facilitates host-to-host circulation and adaptation to man.

The objective of research was to analyse the potential of Normalized Difference Vegetation Index (NDVI) maps from satellite images, yield maps and grapevine fertility and load variables to delineate zones with different wine grape properties for selective harvesting. Two vineyard blocks located in NE Spain (Cabernet Sauvignon and Syrah) were analysed. The NDVI was computed from a Quickbird-2 multi-spectral image at veraison (July 2005). Yield data was acquired by means of a yield monitor during September 2005. Other variables, such as the number of buds, number of shoots, number of wine grape clusters and weight of 100 berries were sampled in a 10 rows × 5 vines pattern and used as input variables, in combination with the NDVI, to define the clusters as alternative to yield maps. Two days prior to the harvesting, grape samples were taken. The analysed variables were probable alcoholic degree, pH of the juice, total acidity, total phenolics, colour, anthocyanins and tannins. The input variables, alone or in combination, were clustered (2 and 3 Clusters) by using the ISODATA algorithm, and an analysis of variance and a multiple rang test were performed. The results show that the zones derived from the NDVI maps are more effective to differentiate grape maturity and quality variables than the zones derived from the yield maps. The inclusion of other grapevine fertility and load variables did not improve the results.

The objective of research was to analyse the potential of Normalized Difference Vegetation Index (NDVI) maps from satellite images, yield maps and grapevine fertility and load variables to delineate zones with different wine grape properties for selective harvesting. Two vineyard blocks located in NE Spain (Cabernet Sauvignon and Syrah) were analysed. The NDVI was computed from a Quickbird-2 multi-spectral image at veraison (July 2005). Yield data was acquired by means of a yield monitor during September 2005. Other variables, such as the number of buds, number of shoots, number of wine grape clusters and weight of 100 berries were sampled in a 10 rows × 5 vines pattern and used as input variables, in combination with the NDVI, to define the clusters as alternative to yield maps. Two days prior to the harvesting, grape samples were taken. The analysed variables were probable alcoholic degree, pH of the juice, total acidity, total phenolics, colour, anthocyanins and tannins. The input variables, alone or in combination, were clustered (2 and 3 Clusters) by using the ISODATA algorithm, and an analysis of variance and a multiple rang test were performed. The results show that the zones derived from the NDVI maps are more effective to differentiate grape maturity and quality variables than the zones derived from the yield maps. The inclusion of other grapevine fertility and load variables did not improve the results.

Anticuerpo anti-Dectin-1 humano, hibridoma productor de dicho anticuerpo y sus aplicaciones. La presente invención consiste en el hibridoma MGD3 y el anticuerpo monocional producido por él (denominado también MGD3), que reconoce de forma especifica al receptor de membrana Decti-1 humano. El anticuerpo MGD3 es capaz de inhibir la unión de Dectin-1 a su ligando natural, los -glucanos que son componentes de la pared de los hongos. Además, bloquea específicamente la unión a Candida albicans así como la secreción de citoquinas inducidas por la misma. El anticuerpo MGD3 obtenido permite la detección in vitro, ex vivo e in vivo de la proteína humana Dectin-1. Del mismo modo, puede emplearse para detectar el nivel de expresión del receptor en leucocitos humanos y para definir el papel de este receptor en la patogenia de enfermedades autoinmunes y de condiciones caracterizadas por inflamación crónica así como para conocer el efecto terapéutico de distintos agentes anti-inflamatorios e inmunosupresores. Además, el anticuerpo MGD3 podría tener potencial terapéutico en enfermedades inflamatorias crónicas.

[EN] The invention relates to hybridoma MGD3 and the monoclonal antibody produced thereby (also called MGD3), which specifically recognises the human Dectin-1 membrane receptor. Antibody MGD3 is capable of inhibiting the binding of Dectin-1 to the natural ligand thereof, the ss-glucans that are components of the fungal wall. In addition, the aforementioned antibody specifically blocks binding to Candida albicans and the secretion of cytokines induced thereby. The MGD3 antibody obtained enables in vitro, ex vivo and in vivo detection of the human protein Dectin-1. Similarly, said antibody can be used to detect the level of expression of the receptor in human leukocytes, to define the role of this receptor in the pathogenesis of autoimmune illnesses and of conditions characterised by chronic inflammation and to ascertain the therapeutic effect of different anti-inflammatory and immunosuppressor agents. In addition, the MGD3 antibody may have therapeutic potential in chronic inflammatory illnesses., [ES] La invención relaciona al hibridoma MGD3 y al anticuerpo monoclonal producido de tal modo (también llamado MGD3), que reconoce específicamente el receptor humano de la membrana Dectin-1. El anticuerpo MGD3 es capaz de inhibir el atascamiento de Dectin-1 al ligand natural de eso, los ss-glucanos los componentes de ese son de la pared fúngica. Además, del anticuerpo los bloques mencionados específicamente que ataban a la Candida albicans y la secreción de citoquinas indujeron de tal modo. El anticuerpo MGD3 obtenido posibilita in vitro, detección viva e in vivo ex de la proteína humana Dectin-1. Semejantemente, el bote dicho del anticuerpo sea utilizado detectar el nivel de expresión del receptor en leucocitos humanos, definir el papel de este receptor en la patogénesis de enfermedades autoinmunes y de las condiciones caracterizadas por la inflamación crónica y determinar el efecto terapéutico de los diferentes agentes antiinflamatorios y del immunosuppressor. Además, el anticuerpo MGD3 puede tener potencial terapéutico en enfermedades inflamatorias crónicas.

Adenoviruses (Ads) are important human pathogens and valuable gene delivery vehicles. We report here the crystal structure of the species B Ad11 knob complexed with the Ad11-binding region of its receptor CD46. The conformation of bound CD46 differs profoundly from its unbound state, with the bent surface structure straightened into an elongated rod. This mechanism of interaction is likely to be conserved among many pathogens that target CD46 or related molecules.

Vaccinia virus enters cells by endocytosis and via a membrane fusion mechanism mediated by viral envelope protein complexes. While several proteins have been implicated in the entry/fusion event, there is no direct proof for fusogenic activity of any viral protein in heterologous systems. Transient coexpression of A17 and A27 in mammalian cells led to syncytia formation in a pH-dependent manner, as ascertained by confocal fluorescent immunomicroscopy. The pH-dependent fusion activity was identified to reside in A17 amino-terminal ectodomain after overexpression in insect cells using recombinant baculoviruses. Through the use of A17 ectodomain deletion mutants, it was found that the domain important for fusion spanned between residues 18 and 34. To further characterize A17–A27 fusion activity in mammalian cells, 293T cell lines stably expressing A17, A27 or coexpressing both proteins were generated using lentivectors. A27 was exposed on the cell surface only when A17 was coexpressed. In addition, pH-dependent fusion activity was functionally demonstrated in mammalian cells by cytoplasmic transfer of fluorescent proteins, only when A17 and A27 were coexpressed. Bioinformatic tools were used to compare the putative A17–A27 protein complex with well-characterized fusion proteins. Finally, all experimental evidence was integrated into a working model for A17–A27-induced pH-dependent cell-to-cell fusion.

[ES] Se analiza la respuesta hidrológica de la cuenca y las subcuencas del río lsábena (Cuenca del Ebro, Pre-Pirineo). Los datos de partida los proporcionan un estudio pluviométrico de detalle y las medidas de caudal en el periodo abril 2000 - abril 2002. La modelización hidrológica se ha realizado mediante el modelo del Número de Curva calibrado. Los resultados indican una estacionalldad marcada de las crecidas, con máxlmos en otoño y primavera. las subcuencas de la parte alta aportaron el 80% de la escorrentía durante crecidas (60% desde la subcuenca de Cabecera y 20% desde la de Villacarli). En eventos tormentosos locales y convectivos los aportes máximos se desplazaron hacia el tramo medio, concretamente hacia las subcuencas de Vlllacarll y Carrasquero. las cuencas septentrionales presen­taron tiempos base superiores a 10 horas y coeficientes de escorrentía del 15%, mientras que en las subcuencas de la parte media los valores fueron de 8 h y del 4%, respectivamente. El estudio constata que la modelización hidrológica con el método del Número de Curva conlleva notables errores en estudios de crecidas de bajo periodo de retorno y que resulta de vital importancia calibrar alguno de sus parámetros con medidas de campo, como por ejemplo el caudal pico.

[ES] Este trabajo analiza la variabilidad espacial y temporal de la precipitación en la cuenca del río lsábena (Cuenca del Ebro, Prepirineo aragonés) durante los años 2000 y 2001. También estudia los errores en la estimación de la precipitación según la fuente de datos y el tipo de interpolación. El estudio se rea­lizó a partir de datos obtenidos mediante 26 pluviómetros totalizadores y 3 pluviógrafos registradores distribuidos en las distintas subcuencas. En la cabecera de la cuenca la precipitación anual fue mayor, pero en la parte media de la cuenca las tormentas fueron más intensas, y con un desfase temporal medio de -50 minutos. La precipitación por episodio siguió un gradiente positivo en sentido sur-norte ( 1,84% /km). Se presentan los polinomios derivados para estimar el valor integrado de precipitación para cada subcuenca a partir de un solo pluviómetro, con coeficientes de ajuste entre el 70% y el 90%. Asimismo se detectó una infraestimación media del 13% en la precipitación a par

[ES] Se analiza la respuesta hidrológica de la cuenca y las subcuencas del río lsábena (Cuenca del Ebro, Pre-Pirineo). Los datos de partida los proporcionan un estudio pluviométrico de detalle y las medidas de caudal en el periodo abril 2000 - abril 2002. La modelización hidrológica se ha realizado mediante el modelo del Número de Curva calibrado. Los resultados indican una estacionalldad marcada de las crecidas, con máxlmos en otoño y primavera. las subcuencas de la parte alta aportaron el 80% de la escorrentía durante crecidas (60% desde la subcuenca de Cabecera y 20% desde la de Villacarli). En eventos tormentosos locales y convectivos los aportes máximos se desplazaron hacia el tramo medio, concretamente hacia las subcuencas de Vlllacarll y Carrasquero. las cuencas septentrionales presen­taron tiempos base superiores a 10 horas y coeficientes de escorrentía del 15%, mientras que en las subcuencas de la parte media los valores fueron de 8 h y del 4%, respectivamente. El estudio constata que la modelización hidrológica con el método del Número de Curva conlleva notables errores en estudios de crecidas de bajo periodo de retorno y que resulta de vital importancia calibrar alguno de sus parámetros con medidas de campo, como por ejemplo el caudal pico.

[ES] Este trabajo analiza la variabilidad espacial y temporal de la precipitación en la cuenca del río lsábena (Cuenca del Ebro, Prepirineo aragonés) durante los años 2000 y 2001. También estudia los errores en la estimación de la precipitación según la fuente de datos y el tipo de interpolación. El estudio se rea­lizó a partir de datos obtenidos mediante 26 pluviómetros totalizadores y 3 pluviógrafos registradores distribuidos en las distintas subcuencas. En la cabecera de la cuenca la precipitación anual fue mayor, pero en la parte media de la cuenca las tormentas fueron más intensas, y con un desfase temporal medio de -50 minutos. La precipitación por episodio siguió un gradiente positivo en sentido sur-norte ( 1,84% /km). Se presentan los polinomios derivados para estimar el valor integrado de precipitación para cada subcuenca a partir de un solo pluviómetro, con coeficientes de ajuste entre el 70% y el 90%. Asimismo se detectó una infraestimación media del 13% en la precipitación a par

[ES] Se analiza la respuesta hidrológica de la cuenca y las subcuencas del río lsábena (Cuenca del Ebro, Pre-Pirineo). Los datos de partida los proporcionan un estudio pluviométrico de detalle y las medidas de caudal en el periodo abril 2000 - abril 2002. La modelización hidrológica se ha realizado mediante el modelo del Número de Curva calibrado. Los resultados indican una estacionalldad marcada de las crecidas, con máxlmos en otoño y primavera. las subcuencas de la parte alta aportaron el 80% de la escorrentía durante crecidas (60% desde la subcuenca de Cabecera y 20% desde la de Villacarli). En eventos tormentosos locales y convectivos los aportes máximos se desplazaron hacia el tramo medio, concretamente hacia las subcuencas de Vlllacarll y Carrasquero. las cuencas septentrionales presen­taron tiempos base superiores a 10 horas y coeficientes de escorrentía del 15%, mientras que en las subcuencas de la parte media los valores fueron de 8 h y del 4%, respectivamente. El estudio constata que la modelización hidrológica con el método del Número de Curva conlleva notables errores en estudios de crecidas de bajo periodo de retorno y que resulta de vital importancia calibrar alguno de sus parámetros con medidas de campo, como por ejemplo el caudal pico.

[ES] Se analiza la respuesta hidrológica de la cuenca y las subcuencas del río lsábena (Cuenca del Ebro, Pre-Pirineo). Los datos de partida los proporcionan un estudio pluviométrico de detalle y las medidas de caudal en el periodo abril 2000 - abril 2002. La modelización hidrológica se ha realizado mediante el modelo del Número de Curva calibrado. Los resultados indican una estacionalldad marcada de las crecidas, con máxlmos en otoño y primavera. las subcuencas de la parte alta aportaron el 80% de la escorrentía durante crecidas (60% desde la subcuenca de Cabecera y 20% desde la de Villacarli). En eventos tormentosos locales y convectivos los aportes máximos se desplazaron hacia el tramo medio, concretamente hacia las subcuencas de Vlllacarll y Carrasquero. las cuencas septentrionales presen­taron tiempos base superiores a 10 horas y coeficientes de escorrentía del 15%, mientras que en las subcuencas de la parte media los valores fueron de 8 h y del 4%, respectivamente. El estudio constata que la modelización hidrológica con el método del Número de Curva conlleva notables errores en estudios de crecidas de bajo periodo de retorno y que resulta de vital importancia calibrar alguno de sus parámetros con medidas de campo, como por ejemplo el caudal pico.

[ES] Se analiza la respuesta hidrológica de la cuenca y las subcuencas del río lsábena (Cuenca del Ebro, Pre-Pirineo). Los datos de partida los proporcionan un estudio pluviométrico de detalle y las medidas de caudal en el periodo abril 2000 - abril 2002. La modelización hidrológica se ha realizado mediante el modelo del Número de Curva calibrado. Los resultados indican una estacionalldad marcada de las crecidas, con máxlmos en otoño y primavera. las subcuencas de la parte alta aportaron el 80% de la escorrentía durante crecidas (60% desde la subcuenca de Cabecera y 20% desde la de Villacarli). En eventos tormentosos locales y convectivos los aportes máximos se desplazaron hacia el tramo medio, concretamente hacia las subcuencas de Vlllacarll y Carrasquero. las cuencas septentrionales presen­taron tiempos base superiores a 10 horas y coeficientes de escorrentía del 15%, mientras que en las subcuencas de la parte media los valores fueron de 8 h y del 4%, respectivamente. El estudio constata que la modelización hidrológica con el método del Número de Curva conlleva notables errores en estudios de crecidas de bajo periodo de retorno y que resulta de vital importancia calibrar alguno de sus parámetros con medidas de campo, como por ejemplo el caudal pico.

[ES] Este trabajo analiza la variabilidad espacial y temporal de la precipitación en la cuenca del río lsábena (Cuenca del Ebro, Prepirineo aragonés) durante los años 2000 y 2001. También estudia los errores en la estimación de la precipitación según la fuente de datos y el tipo de interpolación. El estudio se rea­lizó a partir de datos obtenidos mediante 26 pluviómetros totalizadores y 3 pluviógrafos registradores distribuidos en las distintas subcuencas. En la cabecera de la cuenca la precipitación anual fue mayor, pero en la parte media de la cuenca las tormentas fueron más intensas, y con un desfase temporal medio de -50 minutos. La precipitación por episodio siguió un gradiente positivo en sentido sur-norte ( 1,84% /km). Se presentan los polinomios derivados para estimar el valor integrado de precipitación para cada subcuenca a partir de un solo pluviómetro, con coeficientes de ajuste entre el 70% y el 90%. Asimismo se detectó una infraestimación media del 13% en la precipitación a par

[ES] Se analiza la respuesta hidrológica de la cuenca y las subcuencas del río lsábena (Cuenca del Ebro, Pre-Pirineo). Los datos de partida los proporcionan un estudio pluviométrico de detalle y las medidas de caudal en el periodo abril 2000 - abril 2002. La modelización hidrológica se ha realizado mediante el modelo del Número de Curva calibrado. Los resultados indican una estacionalldad marcada de las crecidas, con máxlmos en otoño y primavera. las subcuencas de la parte alta aportaron el 80% de la escorrentía durante crecidas (60% desde la subcuenca de Cabecera y 20% desde la de Villacarli). En eventos tormentosos locales y convectivos los aportes máximos se desplazaron hacia el tramo medio, concretamente hacia las subcuencas de Vlllacarll y Carrasquero. las cuencas septentrionales presen­taron tiempos base superiores a 10 horas y coeficientes de escorrentía del 15%, mientras que en las subcuencas de la parte media los valores fueron de 8 h y del 4%, respectivamente. El estudio constata que la modelización hidrológica con el método del Número de Curva conlleva notables errores en estudios de crecidas de bajo periodo de retorno y que resulta de vital importancia calibrar alguno de sus parámetros con medidas de campo, como por ejemplo el caudal pico.

[ES] Este trabajo analiza la variabilidad espacial y temporal de la precipitación en la cuenca del río lsábena (Cuenca del Ebro, Prepirineo aragonés) durante los años 2000 y 2001. También estudia los errores en la estimación de la precipitación según la fuente de datos y el tipo de interpolación. El estudio se rea­lizó a partir de datos obtenidos mediante 26 pluviómetros totalizadores y 3 pluviógrafos registradores distribuidos en las distintas subcuencas. En la cabecera de la cuenca la precipitación anual fue mayor, pero en la parte media de la cuenca las tormentas fueron más intensas, y con un desfase temporal medio de -50 minutos. La precipitación por episodio siguió un gradiente positivo en sentido sur-norte ( 1,84% /km). Se presentan los polinomios derivados para estimar el valor integrado de precipitación para cada subcuenca a partir de un solo pluviómetro, con coeficientes de ajuste entre el 70% y el 90%. Asimismo se detectó una infraestimación media del 13% en la precipitación a par

[ES] Este trabajo analiza la variabilidad espacial y temporal de la precipitación en la cuenca del río lsábena (Cuenca del Ebro, Prepirineo aragonés) durante los años 2000 y 2001. También estudia los errores en la estimación de la precipitación según la fuente de datos y el tipo de interpolación. El estudio se rea­lizó a partir de datos obtenidos mediante 26 pluviómetros totalizadores y 3 pluviógrafos registradores distribuidos en las distintas subcuencas. En la cabecera de la cuenca la precipitación anual fue mayor, pero en la parte media de la cuenca las tormentas fueron más intensas, y con un desfase temporal medio de -50 minutos. La precipitación por episodio siguió un gradiente positivo en sentido sur-norte ( 1,84% /km). Se presentan los polinomios derivados para estimar el valor integrado de precipitación para cada subcuenca a partir de un solo pluviómetro, con coeficientes de ajuste entre el 70% y el 90%. Asimismo se detectó una infraestimación media del 13% en la precipitación a par