Molecular dynamics modelling of the interaction of a synthetic zinc-finger miniprotein with DNA

We report the modelling of the DNA complex of an artificial miniprotein composed of two zinc finger modules and an AT-hook linking peptide. The computational study provides for the first time a structural view of these types of complexes, dissecting interactions that are key to modulate their stability. The relevance of these interactions was validated experimentally. These results confirm the potential of this type of computational approach for studying peptide–DNA complexes and suggest that they could be very useful for the rational design of non-natural, DNA binding miniproteinsThis work has received financial support from Spanish grants (IJC2019-040358-I funded by MCIN/AEI/10.13039/501100011033 to J. R., PID2019-108624RB-I00 funded by MCIN/AEI/10.13039/501100011033 to J. L. M. and RTI2018-096704-B-100 and PID2021-122478NB-I00 to M. O.), the Consellería de Cultura, Educación e Ordenación Universitaria (Grants 2015-CP082, ED431C-2017/19, ED431C-2021/ 25 and ED431G 2019/03: Centro Singular de Investigación de Galicia accreditation 2019–2022 to J. L. M.) and the European Union (European Regional Development Fund-ERDF corresponding to the multiannual financial framework 2014–2020 to J. L. M.). This work was also supported by the BioExcel-2. Centre of Excellence for Computational Biomolecular Research” (823830, M. O.) and the Instituto de Salud Carlos III–Instituto Nacional de Bioinformática (ISCIII PT 17/0009/0007 co-funded by the Fondo Europeo de Desarrollo Regional, M. O.)S

Craniopharyngioma in the elderly: a multicenter and nationwide study in Spain

[Abstract] Background: Craniopharyngioma (CP) is a rare tumor in the elderly whose clinical features and prognosis are not well known in this population. Aim: To evaluate the clinicopathological features and therapeutic outcomes of CP diagnosed in the elderly. Patients and Methods: This was a retrospective, multicenter, national study of CP patients diagnosed over the age of 65 years and surgically treated. Results: From a total of 384 adult CP patients, we selected 53 (13.8%) patients (27 women [50.9%], mean age 72.3 ± 5.1 years [range 65–83 years]) diagnosed after the age of 65 years. The most common clinical symptoms were visual field defects (71.2%) followed by headache (45.3%). The maximum tumor diameter was 2.9 ± 1.1 cm. In most patients, the tumor was suprasellar (96.2%) and mixed (solid-cystic) (58.5%). The surgical approach most commonly used was transcranial surgery (52.8%), and more than half of the patients (54.7%) underwent subtotal resection (STR). Adamantinomatous CP and papillary CP were present in 51 and 45.1%, respectively, with mixed forms in the remaining. Surgery was accompanied by an improvement in visual field defects and in headaches; however, pituitary hormonal hypofunction increased, mainly at the expense of an increase in the prevalence of diabetes insipidus (DI) (from 3.9 to 69.2%). Near-total resection (NTR) was associated with a higher prevalence of DI compared with subtotal resection (87.5 vs. 53.6%, p = 0.008). Patients were followed for 46.7 ± 40.8 months. The mortality rate was 39.6% with a median survival time of 88 (95% CI: 57–118) months. DI at last visit was associated with a lower survival. Conclusion: CP diagnosed in the elderly shows a similar distribution by sex and histologic forms than that diagnosed at younger ages. At presentation, visual field alterations and headaches are the main clinical symptoms which improve substantially with surgery. However, surgery, mainly NTR, is accompanied by worsening of pituitary function, especially DI, which seems to be a predictor of mortality in this population

Effects of intubation timing in patients with COVID-19 throughout the four waves of the pandemic : a matched analysis

The primary aim of our study was to investigate the association between intubation timing and hospital mortality in critically ill patients with COVID-19-associated respiratory failure. We also analysed both the impact of such timing throughout the first four pandemic waves and the influence of prior non-invasive respiratory support on outcomes. This is a secondary analysis of a multicentre, observational and prospective cohort study that included all consecutive patients undergoing invasive mechanical ventilation due to COVID-19 from across 58 Spanish intensive care units (ICU) participating in the CIBERESUCICOVID project. The study period was between 29 February 2020 and 31 August 2021. Early intubation was defined as that occurring within the first 24 h of intensive care unit (ICU) admission. Propensity score (PS) matching was used to achieve balance across baseline variables between the early intubation cohort and those patients who were intubated after the first 24 h of ICU admission. Differences in outcomes between early and delayed intubation were also assessed. We performed sensitivity analyses to consider a different timepoint (48 h from ICU admission) for early and delayed intubation. Of the 2725 patients who received invasive mechanical ventilation, a total of 614 matched patients were included in the analysis (307 for each group). In the unmatched population, there were no differences in mortality between the early and delayed groups. After PS matching, patients with delayed intubation presented higher hospital mortality (27.3% versus 37.1%, p =0.01), ICU mortality (25.7% versus 36.1%, p=0.007) and 90-day mortality (30.9% versus 40.2%, p=0.02) when compared to the early intubation group. Very similar findings were observed when we used a 48-hour timepoint for early or delayed intubation. The use of early intubation decreased after the first wave of the pandemic (72%, 49%, 46% and 45% in the first, second, third and fourth wave, respectively; first versus second, third and fourth waves p<0.001). In both the main and sensitivity analyses, hospital mortality was lower in patients receiving high-flow nasal cannula (n=294) who were intubated earlier. The subgroup of patients undergoing NIV (n=214) before intubation showed higher mortality when delayed intubation was set as that occurring after 48 h from ICU admission, but not when after 24 h. In patients with COVID-19 requiring invasive mechanical ventilation, delayed intubation was associated with a higher risk of hospital mortality. The use of early intubation significantly decreased throughout the course of the pandemic. Benefits of such an approach occurred more notably in patients who had received high-flow nasal cannul

Prognostic implications of comorbidity patterns in critically ill COVID-19 patients: A multicenter, observational study

Background The clinical heterogeneity of COVID-19 suggests the existence of different phenotypes with prognostic implications. We aimed to analyze comorbidity patterns in critically ill COVID-19 patients and assess their impact on in-hospital outcomes, response to treatment and sequelae. Methods Multicenter prospective/retrospective observational study in intensive care units of 55 Spanish hospitals. 5866 PCR-confirmed COVID-19 patients had comorbidities recorded at hospital admission; clinical and biological parameters, in-hospital procedures and complications throughout the stay; and, clinical complications, persistent symptoms and sequelae at 3 and 6 months. Findings Latent class analysis identified 3 phenotypes using training and test subcohorts: low-morbidity (n=3385; 58%), younger and with few comorbidities; high-morbidity (n=2074; 35%), with high comorbid burden; and renal-morbidity (n=407; 7%), with chronic kidney disease (CKD), high comorbidity burden and the worst oxygenation profile. Renal-morbidity and high-morbidity had more in-hospital complications and higher mortality risk than low-morbidity (adjusted HR (95% CI): 1.57 (1.34-1.84) and 1.16 (1.05-1.28), respectively). Corticosteroids, but not tocilizumab, were associated with lower mortality risk (HR (95% CI) 0.76 (0.63-0.93)), especially in renal-morbidity and high-morbidity. Renal-morbidity and high-morbidity showed the worst lung function throughout the follow-up, with renal-morbidity having the highest risk of infectious complications (6%), emergency visits (29%) or hospital readmissions (14%) at 6 months (p<0.01). Interpretation Comorbidity-based phenotypes were identified and associated with different expression of in-hospital complications, mortality, treatment response, and sequelae, with CKD playing a major role. This could help clinicians in day-to-day decision making including the management of post-discharge COVID-19 sequelae. Copyright (C) 2022 The Author(s). Published by Elsevier Ltd

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

The evolution of the ventilatory ratio is a prognostic factor in mechanically ventilated COVID-19 ARDS patients

Background: Mortality due to COVID-19 is high, especially in patients requiring mechanical ventilation. The purpose of the study is to investigate associations between mortality and variables measured during the first three days of mechanical ventilation in patients with COVID-19 intubated at ICU admission. Methods: Multicenter, observational, cohort study includes consecutive patients with COVID-19 admitted to 44 Spanish ICUs between February 25 and July 31, 2020, who required intubation at ICU admission and mechanical ventilation for more than three days. We collected demographic and clinical data prior to admission; information about clinical evolution at days 1 and 3 of mechanical ventilation; and outcomes. Results: Of the 2,095 patients with COVID-19 admitted to the ICU, 1,118 (53.3%) were intubated at day 1 and remained under mechanical ventilation at day three. From days 1 to 3, PaO2/FiO2 increased from 115.6 [80.0-171.2] to 180.0 [135.4-227.9] mmHg and the ventilatory ratio from 1.73 [1.33-2.25] to 1.96 [1.61-2.40]. In-hospital mortality was 38.7%. A higher increase between ICU admission and day 3 in the ventilatory ratio (OR 1.04 [CI 1.01-1.07], p = 0.030) and creatinine levels (OR 1.05 [CI 1.01-1.09], p = 0.005) and a lower increase in platelet counts (OR 0.96 [CI 0.93-1.00], p = 0.037) were independently associated with a higher risk of death. No association between mortality and the PaO2/FiO2 variation was observed (OR 0.99 [CI 0.95 to 1.02], p = 0.47). Conclusions: Higher ventilatory ratio and its increase at day 3 is associated with mortality in patients with COVID-19 receiving mechanical ventilation at ICU admission. No association was found in the PaO2/FiO2 variation

Estudio de prefactibilidad para la implementación de servicios exequiales para mascotas en la ciudad de Bogotá.

Elaborar el estudio de prefactibilidad, recopilando información y tomando en cuenta diferentes variables que permitan realizar un análisis preliminar y de esta manera minimizar el riesgo para la creación de una empresa de servicios exequiales de mascotas ubicada en la ciudad de Bogotá.Debido al desarrollo y alto crecimiento de servicios fúnebres para mascotas en Colombia, las nuevas tendencias en el mercado de las mascotas permiten generar una gran oportunidad a las empresas funerarias en diversificar su portafolio de servicios y fortalecer sus ventas en previsión exequial a través de la atención profesionalizada en servicios funerarios para mascotas. Esta línea de negocio puede funcionar también como una estrategia de venta para las afiliaciones exequiales mediante la aceptación de la mascota como un miembro de la familia, lo que generará nuevas afiliaciones de previsión exequial y mayor fidelización para con la empresa. Es por esto por lo que, se necesita de mucha promoción, sensibilidad, concientización, creatividad e innovación para poder vender al cliente, por lo tanto, se recomienda utilizar estrategias de promoción incentivando a la compra de estos servicios a través de beneficios adicionales para el comprador. A corto plazo para generar reconocimiento en la comunidad y el perfil de nuestro comprador, se deberá ejecutar el plan de promoción informativo, con el cual buscara los clientes objetivos, utilizando estrategias en Marketing digital y alianzas estratégicas. Las redes sociales juegan un papel importante, por medio de ellas se tendrá actualizado a los clientes, se generarán mecanismos para aumentar la comunidad virtual y así poder dar a conocer la marca. Es importante la constante innovación y participación de la nueva línea de negocio en todo el ciclo de vida de la mascota (servicios y productos que pueda utilizar en vida y en su muerte), para romper el paradigma que se está vendiendo un seguro de muerte, sino al contrario, se está vendiendo una protección y bienestar para la familia, que incluye a su mascota.Due to the development and high growth of funeral services for pets in Colombia, the new trends in the pet market allow a great opportunity for funeral companies to diversify their portfolio of services and strengthen their sales in funeral provision through care Professionalized in funeral services for pets. This line of business can also function as a sales strategy for funeral affiliations by accepting the pet as a member of the family, which will generate new funeral funeral affiliations and greater loyalty to the company. This is why, it takes a lot of promotion, sensitivity, awareness, creativity and innovation to be able to sell to the client, therefore, it is recommended to use promotion strategies encouraging the purchase of these services through additional benefits for the buyer. In the short term to generate recognition in the community and the profile of our buyer, the informative promotion plan must be executed, with which it will search for the target clients, using strategies in digital Marketing and strategic alliances. Social networks play an important role, through them customers will be updated, mechanisms will be generated to increase the virtual community and thus be able to publicize the brand. The constant innovation and participation of the new business line throughout the pet's life cycle (services and products that can be used in life and in death) is important to break the paradigm that death insurance is being sold, on the contrary, a protection and well-being for the family is being sold, which includes your pet

Estudio de prefactibilidad para la implementación de servicios exequiales para mascotas en la ciudad de Bogotá.

Elaborar el estudio de prefactibilidad, recopilando información y tomando en cuenta diferentes variables que permitan realizar un análisis preliminar y de esta manera minimizar el riesgo para la creación de una empresa de servicios exequiales de mascotas ubicada en la ciudad de Bogotá.Debido al desarrollo y alto crecimiento de servicios fúnebres para mascotas en Colombia, las nuevas tendencias en el mercado de las mascotas permiten generar una gran oportunidad a las empresas funerarias en diversificar su portafolio de servicios y fortalecer sus ventas en previsión exequial a través de la atención profesionalizada en servicios funerarios para mascotas. Esta línea de negocio puede funcionar también como una estrategia de venta para las afiliaciones exequiales mediante la aceptación de la mascota como un miembro de la familia, lo que generará nuevas afiliaciones de previsión exequial y mayor fidelización para con la empresa. Es por esto por lo que, se necesita de mucha promoción, sensibilidad, concientización, creatividad e innovación para poder vender al cliente, por lo tanto, se recomienda utilizar estrategias de promoción incentivando a la compra de estos servicios a través de beneficios adicionales para el comprador. A corto plazo para generar reconocimiento en la comunidad y el perfil de nuestro comprador, se deberá ejecutar el plan de promoción informativo, con el cual buscara los clientes objetivos, utilizando estrategias en Marketing digital y alianzas estratégicas. Las redes sociales juegan un papel importante, por medio de ellas se tendrá actualizado a los clientes, se generarán mecanismos para aumentar la comunidad virtual y así poder dar a conocer la marca. Es importante la constante innovación y participación de la nueva línea de negocio en todo el ciclo de vida de la mascota (servicios y productos que pueda utilizar en vida y en su muerte), para romper el paradigma que se está vendiendo un seguro de muerte, sino al contrario, se está vendiendo una protección y bienestar para la familia, que incluye a su mascota.Due to the development and high growth of funeral services for pets in Colombia, the new trends in the pet market allow a great opportunity for funeral companies to diversify their portfolio of services and strengthen their sales in funeral provision through care Professionalized in funeral services for pets. This line of business can also function as a sales strategy for funeral affiliations by accepting the pet as a member of the family, which will generate new funeral funeral affiliations and greater loyalty to the company. This is why, it takes a lot of promotion, sensitivity, awareness, creativity and innovation to be able to sell to the client, therefore, it is recommended to use promotion strategies encouraging the purchase of these services through additional benefits for the buyer. In the short term to generate recognition in the community and the profile of our buyer, the informative promotion plan must be executed, with which it will search for the target clients, using strategies in digital Marketing and strategic alliances. Social networks play an important role, through them customers will be updated, mechanisms will be generated to increase the virtual community and thus be able to publicize the brand. The constant innovation and participation of the new business line throughout the pet's life cycle (services and products that can be used in life and in death) is important to break the paradigm that death insurance is being sold, on the contrary, a protection and well-being for the family is being sold, which includes your pet

SUMOylation of p53 mediates interferon activities

There is growing evidence that many host proteins involved in innate and intrinsic immunity are regulated by SUMOylation, and that SUMO contributes to the regulatory process that governs the initiation of the type I interferon (IFN) response. The tumor suppressor p53 is a modulator of the IFN response that plays a role in virus-induced apoptosis and in IFN-induced senescence. Here we demonstrate that IFN treatment increases the levels of SUMOylated p53 and induces cellular senescence through a process that is partially dependent upon SUMOylation of p53. Similarly, we show that vesicular stomatitis virus (VSV) infection induces p53 SUMOylation, and that this modification favors the control of VSV replication. Thus, our study provides evidence that IFN signaling induces p53 SUMOylation, which results in the activation of a cellular senescence program and contributes to the antiviral functions of interferon