Plitidepsin Is a Broad-Spectrum Antiviral Shaping the Cell Proteostatic Balance

Background: Different viruses employ similar pathways for replication, revealing key intracellular hotspots to target with host-directed therapies and achieve a broad-spectrum antiviral activity. Plitidepsin is a clinically approved antitumoral agent that blocks the elongation factor eEF1A required for protein translation. This drug counteracts SARS-CoV-2 replication and shows a favorable safety profile in COVID-19 patients. Yet, the precise antiviral mechanism of action of plitidepsin remains unknown. Method(s): Here we used a deep quantitative proteomic analysis to measure the impact of plitidepsin on the proteome of SARS-CoV-2-infected Vero E6 cells. This was complemented with transmission electron microscopy assays, which unraveled the subcellular and morphological changes associated to plitidepsin treatment. In addition, we performed functional in vitro assays to dissect the antiviral activity of plitidepsin against SARS-CoV-2 and other viruses. Result(s): We found that this drug inhibited the synthesis of all SARS-CoV-2 proteins in a dose-dependent manner. These included the R1AB polyproteins, which facilitate the synthesis of non-structural proteins involved in the formation of double membrane vesicles (DMV) required for viral replication. Plitidepsin reduced DMV formation and the morphogenesis of new viruses, having a greater impact on viral than on host proteins. Less than 14% of the cellular proteome was significantly affected by plitidepsin, inducing the up-regulation of key molecules associated with protein biosynthesis, such as the translation initiation factors eIF4A2 and eIF2S3. Therefore, plitidepsin induced a compensatory state that rescued protein translation. This proteostatic response explains how cells preserve the cellular proteome after treatment with a translation inhibitor such as plitidepsin. In addition, it suggests that plitidepsin could inhibit other RNA-dependent and non-integrated DNA viruses, as we confirmed in vitro using Zika virus, Hepatitis C virus replicon and Herpes simplex virus. However, the compensatory proteostasis induced by plitidespin also explains why this drug failed to inhibit the replication of integrated DNA proviruses such as HIV-1. Conclusion(s): Unraveling the mechanism of action of host-directed therapies like plitidepsin is imperative to define the indications and antiviral profile of these compounds. This knowledge will be key to develop broad-spectrum treatments and have them ready to deploy when future pandemic viruses break through.

Busqueda De Un Patron Clinico Para La Prediccion De Colonoscopia Patologica En Los Pacientes Participantes De Las 2 Primeras Rondas Del Programa De Deteccion Precoz De Cancer Colorrectal Del Hospital Comarcal De Inca

Introduccion: En el desarrollo de los Programas de Deteccion Precoz de Cancer Colorrectal (PDPCCR), la colonoscopia es la prueba diagnostica final, asi como el principal factor l imitante, este hecho sumado a situaciones como la reciente pandemia por COVID, que puso en minimos el funcionamiento de varios servicios de endoscopia a nivel nacional, hace que nos plantearamos intentar buscar patrones predictivos que a futuro permitan la priorizacion de aquellos pacientes con mayor riesgo de endoscopia patologica dentro del programa de cribado, a fin de agilizar el desarrollo de los mismos. Metodos: Estudio retrospectivo basado en una muestra de datos clinicos y endoscopicos recopilados de las 2 primeras rondas del PDPCCR del Area de la Tramuntana-Hospital Comarcal de Inca (Mallorca). Resultados: Se encontro asociacion entre la variabilidad de la edad, el sexo y los valores cuantitativos de la sangre oculta en heces inmunoquimica (SOHi) con el riesgo de hallazgos endoscopicos, asi como la toma de antiinflamatorios no esteroideos (AINES) y acido acetilsalicilico (AAS) como factores protectores. Conclusiones: Hemos podido replicar los resultados de otros estudios, lo que fortalece nuestro programa de cribado, y nos sirve de base para obtener prospectivamente modelos de riesgo mas precisos.Copyright © 2023 Elsevier Espana, S.L.U. Todos los derechos reservados.

Post-Covid-19 Irritable Bowel Syndrome

Background and aim: The long-term consequences of COVID- 19 infection on the gastrointestinal tract remain unclear. Here we aimed to evaluate the prevalence of gastrointestinal symptoms and post-COVID-19 disorders of gut-brain interaction (DGBI) after hospitalization for SARS-CoV-2 infection. Material(s) and Method(s): GI-COVID19 is a prospective, multicenter, controlled study. Patients with and without COVID-19 diagnosis were evaluated upon hospital admission and after 1, 6, and 12 months post-hospitalization. Gastrointestinal symptoms, anxiety, and depression were assessed using validated questionnaires, namely the Gastrointestinal Symptoms Rating Scale (GSRS), the Hanxiety and Depression Scale (HADS) and the Rome IV Diagnostic Questionnaire for Functional Gastrointestinal Disorders in Adults. Result(s): The study included 2183 hospitalized patients. The primary analysis included a total of 883 patients (614 COVID-19 patients and 269 controls) due to the exclusion of patients with pre-existing gastrointestinal symptoms and/or surgery. At enrollment, gastrointestinal symptoms were more frequent among COVID-19 patients than in the control group (59.3% vs. 39.7%, P<0.001). At the 12-month follow- up, constipation and hard stools were significantly more prevalent in controls than in COVID-19 patients (16% vs. 9.6%, P=0.019 and 17.7% vs. 10.9%, P=0.011, respectively). Compared to controls, COVID- 19 patients reported higher rates of irritable bowel syndrome (IBS) according to Rome IV criteria: 0.5% vs. 3.2%, P=0.045. Factors significantly associated with IBS diagnosis included history of allergies, chronic intake of proton pump inhibitors, and presence of dyspnea. [Table presented] At the 6-month follow-up, the rate of COVID-19 patients fulfilling the criteria for depression was higher than among controls. Conclusion(s): Compared to controls, hospitalized COVID-19 patients had fewer complaints of constipation and hard stools at 12 months after acute infection. COVID-19 patients had significantly higher rates of IBS than controls. ClinicalTrials.gov number, NCT04691895.Copyright © 2023. Editrice Gastroenterologica Italiana S.r.l.

Developing an assay to distinguish between HIT and VITT antibodies

Introduction Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a rare, but severe side effect after Covid-19 and other vaccinations. First cases of VITT-mimicking antibodies in unvaccinated patients with recurrent thrombosis have been described. Differentiation between heparin-induced thrombocytopenia (HIT) and VITT is difficult in some patients. Widely used enzyme-linked immunoassays (EIA) cannot differentiate between the two, some of them even fail to detect VITT antibodies. So far, differentiation between HIT-like and VITT-like anti-PF4 antibodies can only be performed in specialized laboratories by functional tests using the heparin-induced platelet activation (HIPA) or PF4-induced platelet activation (PIPA) test. We have developed an assay, which can distinguish between HIT and VITT antibodies and can be used in any hospital laboratory. Method Confirming platelet-activation assays (HIPA and PIPA) were performed as described.[1] We defined 3 cohorts: 1) Negative controls (n = 112, including 35 healthy donors from before 2020, 46 clinical patients suspected for HIT but with negative EIA and HIPA and 31 non-thrombotic patients);2) classical HIT-patients with positive EIA and HIPA (n = 121);3) typical VITT patients (n = 63;presenting after vaccination with adenoviral vector-based Covid-19 vaccine and positive EIA and PIPA). Samples were analyzed by an automated coagulation analyzer ACL AcuStar (Werfen / IL Inc., Bedford, MA, USA) using HemosIL AcuStar HIT-IgG(PF4-H) and a prototype of VITT-IgG(PF4) assay according to the manufacturer's protocol. For both assays, raw data was analyzed as relative light units (RLU). Results All VITT samples were positive in the prototype VITT-assay (Fig. 1);only a few (n = 9;14.3 %) also showed weakly positive results in the HIT-assay. On the other hand, most of the HIT samples showed positive results in the HIT-assay (113;93.4 %), 34 of them (30.1 %) also reacted positive in the prototype VITT-assay (12 of them strongly;10.6 %), and three demonstrated an antibody pattern like autoimmune VITT. Negative control samples where all non-reactive in the HITassay and served to adjust the cutoff for the prototype VITT-assay. Conclusion The different reaction pattern of samples of HIT and VITT patients using HemosIL AcuStar HIT-IgG(PF4-H) and a VITT prototype assay was able to distinguish between the two antibody entities for the first time. The combination of assays can facilitate a rapid decision whether heparin may be used for treatment and also identify patients with autoimmune-VITT as a cause of recurrent thrombosis. (Table Presented).

Post-Infection Irritable Bowel Syndrome in Sars-Cov-2 Patients: The Gi-Covid19 Study

Introduction: SARS-CoV-2 infection, known as COVID-19, may lead to persistent gastrointestinal dysfunction resembling aspects of post-infection disorders of gut-brain interaction (DGBI). However, the long-term consequences of COVID-19 on the gastrointestinal tract remain unclear. Aims & Methods: We aimed to evaluate the prevalence of gastrointestinal symptoms and post-infection disorders of gut-brain interaction (DGBI) up to 12 months after hospitalization and the factors associated with their presence. The GI-COVID19 is a prospective, multicenter, controlled study. Patients with and without COVID-19 diagnosis were assessed at hospital admission and followed up after 1, 6, and 12 months to assess gastrointestinal symptoms using the Gastrointestinal Symptoms Rating Scale, the Rome IV Diagnostic Questionnaire for Functional Gastrointestinal Disorders in Adults, and the hospital anxiety and depression scale. ClinicalTrials. gov number, NCT04691895. Result(s): The study included2183 hospitalized patients. After excluding patients with pre-existing gastrointestinal symptoms and/or surgery, a total of 883 patients (614 COVID-19 and 269 controls) were included in the primary analysis, of whom 435 COVID-19 and 188 controls completed 12 months of follow-up. At enrollment, gastrointestinal symptoms occurred more frequently in COVID-19 patients than in the control group (59.3% vs. 39.7%, P<0.001). Symptoms more frequently complained by COVID-19 patients at enrollment were nausea, diarrhea, loose stool, and urgency. At 1-month follow-up evaluation, nausea and acid regurgitation were significantly more prevalent in COVID-19 patients than in the control group (8.7% vs. 1.7%, P=0.015 and 8.4% vs. 2.1%, P=0.006, respectively). At 6 months, COVID-19 patients reported lower rates of flatus (17.6% vs. 19.1%, P=0.024), constipation (8.9% vs. 17.1%, P<0.001) and hard stools (9.6 vs. 17.2%, P=0.030) as compared with the control group. At 12 months, constipation and hard stools were significantly less prevalent in COVID-19 patients than in the control group (9.6% vs. 16%, P=0.019 and 10.9% vs. 17.7%, P=0.011, respectively). COVID-19 patients reported higher rates of DGBI during follow-up compared to controls (Table), although statistically significant differences were found only for irritable bowel syndrome (IBS) according to Rome III criteria (4.4% vs 1.1%, P=0.036) and Rome IV criteria (3.2% vs 0.5%, P=0.045). The rate of COVID-19 patients depressed at 6 months and with anxiety at 12 months was higher compared to controls (4.1% vs 2.7%, P=0.014 and 4.5% vs 1.1%, P=0.088, respectively). Factors significantly associated with IBS diagnosis were anamnestic allergies (OR 10.024, 95% CI 1.766-56.891, P=0.009), chronic intake of proton pump inhibitors (OR 4.816, 95% CI 1.447-16.025, P=0.010) and dyspnea (OR 4.157, 95% CI 1.336-12.934, P=0.014). Conclusion(s): Hospitalized COVID-19 patients complain less constipation and hard stools than control at 12 months after acute infection. COVID-19 patients are also more likely to develop IBS.

TRANSFORMATION OF A SURGICAL CLINIC FOR COVID-19 PATIENTS: EXPERIENCE REPORT

Objective: to report the transformation process of a surgical inpatient unit into a coronavirus patient care unit and the impact on the health of nursing professionals and users of this service, in the opinion of nurses who experienced the process. Method: this is an experience report of nurses of a surgical inpatient unit of a Federal Teaching Hospital in the Midwest region of Brazil. The experience was presented in three thematic axes: 1) Internal restructuring of the surgical hospitalization unit;2) Training, personnel management and distribution of resources;3) Impacts on worker health. Results: the surgical clinic started to have an exclusive team and flow of single care. Pre-surgical patients were discharged from the hospital with no prospect of returning to the planned treatment. Surgical wards were adapted to isolation wards, and equipment facilities for ventilatory care were optimized. The lack of adequate Personal Protective Equipment and work overload were identified as situations that increase the risk of infection in health professionals. Conclusion: the study demonstrated unhealthy conditions that favor the physical exhaustion and mental suffering of nursing professionals. New research in occupational health planning which supports the work of health service managers in future pandemic scenarios in Brazil is suggested. © 2022, Universidade Federal de Santa Catarina. All rights reserved.

Proposal for a Low-Cost Personal Protective Equipment (PPE) to Protect Health Care Professionals in the Fight Against Coronavirus

The global situation caused by a coronavirus in 2020 has become an unprecedented health crisis. Since 1918, during the Spanish flu, Brazilian society haven’t seen a pandemic disease. Among the many individuals exposed to the virus, health care professionals are essential workers who fight the disease. However, due to the lack of previous investments in the public health sector in Brazil, several obstacles are evident, such as the reduced number of Personal Protective Equipment (PPE) available for healthcare professionals who have direct contact with those affected by the disease. Therefore, undergraduate students and professors from UFPA (Universidade Federal do Pará—Federal University of Pará) Biomedical Engineering program course mobilized to create solutions to this problem through the study and production of a low-cost face shield model, which proved to be essential in helping professionals, in addition to engaging several students in the social causes. Similar initiatives like this took place in states in southeastern Brazil, through the making of PPE in 3D printers from PLA (polylactic acid) or ABS (Acrylonitrile Butadiene Styrene) filaments, which can take hours to be ready. Therefore, the present study presents a handmade model to promote higher cost–benefit and shorter manufacturing time. To measure how this PPE proposed here affected the daily work of these professionals, this study performed a feedback form focused on the professionals who used the handmade model. Also, a comparison between the 3D printed versus the handmade model was done. The study showed that 69.5% of the respondents, which were professionals working in the health field, felt more protected with the handmade face shield. Finally, for this research, it is possible to say that the handmade faceshields have proven to be effective, providing confidence when used by the research participants. © 2022, Springer Nature Switzerland AG.

Effect of covid related restrictions on irritable bowel syndrome psychological well-being. a comparative study with ulcerative colitis

Introduction: Patients with Irritable bowel syndrome (IBS) have been reported to have higher anxiety and depression levels than general population, as well as dysfunctional coping skills. The total/ partial lockdown due to COVID19 pandemic has been related with an increased incidence of psychological disorders in general population;however, its effect on IBS patients and patients with other chronic GI-conditions like ulcerative colitis (UC) is not known. Aims & Methods: The main goal of this study was to compare the lockdown effect on mental health and digestive symptoms in patients with IBS, UC and healthy volunteers. During April 2020 (Spanish total lockdown), 30 IBS, 30 UC in remission and 30 healthy volunteers were invited to answer an online survey evaluating stress and anxiety levels by the STAI questionnaire and depression using the Beck questionnaire. IBS patients' symptoms were evaluated with IBS-SSS. After 6 months, in October 2020 (partial lockdown in Spain), the same subjects were contacted, and they were asked to answer again the same online questionnaires. Results: Responses were obtained from 24 IBS (67% women, aged 21-67 years), 23 UC (56% women, aged 35-55 years) and 18 healthy volunteers (68% women, aged 23-70 years). During total lockdown, IBS patients presented higher depression index (11.52±1.6) than UC patients (5.78±0.9 (p=0.008)) and healthy volunteers (4.78±4.3 (p=0.001)). IBS patients also had higher anxiety index (state 29.4±2.5 / trait 26.5±2.5) than healthy volunteers (state 15.6±2.3 (p=0.001) / trait 14.4±1.9 (p=0.002)). After 6 months (during partial lockdown) depression levels increased significatively in IBS patients (from 11.52±1.6 to 16.08±2.22 (p=0.012)), but remained unchanged in healthy subjects and UC patients (from 4.78±4.3 to 3.28±0.6 and from 5.78±0.9 to 5.91±1.17 respectively;p< 0.001 vs IBS). Anxiety scores (both state and trait) remained stable during the partial lockdown (6 months after the total lockdown) in the 3 study groups. The severity of digestive symptoms remained unchanged in IBS patients (scores 283±80 and 285±84, total and partial lockdown, respectively;p=0.837). Conclusion: During the lockdown due to COVID-19 pandemic, IBS patients presented higher depression and anxiety levels than healthy volunteers and patients with UC in remission. Additionally, the depression index worsened in IBS patients during the pandemic, while it remained stable in healthy volunteers and UC patients. During this first phase of the pandemic, the observed changes in psychological traits did not impact on abdominal symptoms, but carefully monitoring of IBS patients in the next months seems to be crucial to evaluate the potential repercussion of worsened mental health due to COVID-19 pandemic in IBS.

Impact of the COVID-19 pandemic on the mortality of the elderly patient with a hip fracture. / Impacto de la pandemia COVID-19 en la mortalidad del paciente anciano con fractura de cadera.

COVID-19 became a threat to the public health system, compromising the health of the population. Patients with hip fractures, due to their age and comorbidity, were high-risk patients in this pandemic. The purpose of this study was to observe how the pandemic affected the management of hip fractures in elderly patients. METHODS: This is a descriptive, retrospective study of all patients over the age of 65 diagnosed with a hip fracture that came to the emergency room of Vall d'Hebron University Hospital in the COVID-19 pandemic period, from the 11th of March to the 24th of April 2020. They were followed up during their hospital stay and 30 days after the fracture. RESULTS: A total of 63 patients were included, 18 (28.6%) of whom had a positive RT-qPCR for COVID-19. Four could not be operated on due to the severity of the disease they presented with upon admission, dying a few days afterwards. Three of these patients had COVID-19. The 83.3% of the patients with positive RT-qPCR presented respiratory symptoms during their hospitalization. The length of hospital stays of patients with a positive RT-qPCR (18.25±8.99 days) was longer than that of patients that were RT-qPCR negative (10.9±4.52 days) (P=.01). In-hospital mortality in operated patients was 20% in patients with a positive RT-qPCR, compared with 2.3% in the group of patients who tested negative (P=.018). Mortality at 30 days was 40% in the group with positive RT-qPCR vs 6.8% in patients not infected by SARS-CoV-2 (P=.002). CONCLUSION: SARS-CoV-2 infection in elderly patients with hip fractures increases both the length of hospital stay, as well as in-hospital and 30-day mortality.

Recommendations for use of antigenic tests in the diagnosis of acute SARS-CoV-2 infection in the second pandemic wave: attitude in different clinical settings.

The high transmissibility of SARS-CoV-2 before and shortly after the onset of symptoms suggests that only diagnosing and isolating symptomatic patients may not be sufficient to interrupt the spread of infection; therefore, public health measures such as personal distancing are also necessary. Additionally, it will be important to detect the newly infected individuals who remain asymptomatic, which may account for 50% or more of the cases. Molecular techniques are the "gold standard" for the diagnosis of SARS-CoV-2 infection. However, the massive use of these techniques has generated some problems. On the one hand, the scarcity of resources (analyzers, fungibles and reagents), and on the other the delay in the notification of results. These two facts translate into a lag in the application of isolation measures among cases and contacts, which favors the spread of the infection. Antigen detection tests are also direct diagnostic methods, with the advantage of obtaining the result in a few minutes and at the very "pointof-care". Furthermore, the simplicity and low cost of these tests allow them to be repeated on successive days in certain clinical settings. The sensitivity of antigen tests is generally lower than that of nucleic acid tests, although their specificity is comparable. Antigenic tests have been shown to be more valid in the days around the onset of symptoms, when the viral load in the nasopharynx is higher. Having a rapid and real-time viral detection assay such as the antigen test has been shown to be more useful to control the spread of the infection than more sensitive tests, but with greater cost and response time, such as in case of molecular tests. The main health institutions such as the WHO, the CDC and the Ministry of Health of the Government of Spain propose the use of antigenic tests in a wide variety of strategies to respond to the pandemic. This document aims to support physicians involved in the care of patients with suspected SC2 infection, in the context of a growing incidence in Spain since September 2020, which already represents the second pandemic wave of COVID-19.