Anesthesia training: Are we doing enough in three years? Cross-sectional study

Introduction: The mínimum number of procedures required to be performed during anesthesia training has not been officially defined in Colombia. Although a number is no guarantee of acquired competencies, it does indicate the level of opportunity offered by the different programs. This study describes the practical training afforded to residents in a graduate anesthesia program in Colombia, and compares its results with international standards. Objective: Describe exposure to procedures performed by residents enrolled in a three-year anesthesia specialization program in Colombia between 2015 and 2020, and compare with the standards proposed by ASCOFAME and ACGME. Methods: Descriptive, cross-sectional study which included residents who did their specialization in a Colombian anesthesia program between 2015 and 2020. Complexity, anesthesia techniques, invasive monitoring and airway approach were described. Finally a descriptive comparison was made with the published references of the Colombian Association of Medical Schools (ASCOFAME) and the Accreditation Council for Graduate Medical Education (ACGME). Results: The results for 10 residents were included. Each resident had a median of 978 cases (IQR 942-1120), corresponding to 25 surgical specialties, the most frequent being general surgery (18%), orthopedics (16%), pediatric surgery (19%), and obstetrics (10.8%). According to the American Society of Anesthesiology (ASA) classification, the majority of patients were ASA II (39.63%) and ASA III (28.4%). Adequate exposure was achieved in 11 of the 15 categories proposed by ACGME and in 6 of the 15 proposed by ASCOFAME. Conclusions: A detailed description of the practice component acquired by the residents during their three years of training was obtained. This baseline provides insight into the national landscape and allows to describe the relationship with international standards.

Introducción: En Colombia no se encuentra oficialmente definido el número mínimo de procedimientos que se deben realizar durante el entrenamiento en anestesiología. Aunque el número no garantiza la adquisición de competencias de la especialidad, sí es un indicador de la oportunidad ofertada por parte de los programas. Este estudio describe el entrenamiento práctico que tienen los médicos residentes en un programa de posgrado de anestesiología en Colombia y compara sus resultados con estándares internacionales. Objetivo: Describir la exposición a procedimientos realizados por los médicos residentes de un programa de especialización en anestesiología de tres años en Colombia, entre 2015 y 2020, y compararlo con los estándares propuestos por ASCOFAME y el ACGME. Métodos: Estudio descriptivo de corte transversal; se incluyeron los residentes que cursaron su programa de especialidad en un programa colombiano de anestesiología entre 2015 y 2020. Se describieron la complejidad, técnicas anestésicas, monitoría invasiva y abordaje de la vía aérea. Finalmente, se compararon los resultados de manera descriptiva con lo referenciado por la Asociación Colombiana de Facultades de Medicina y el Accreditation Council for Graduate Medical Education (ACGME). Resultados: Se incluyeron los resultados de 10 médicos residentes. El número de casos por residente tuvo una mediana de 978 casos (RIQ942-1120), correspondientes a 25 especialidades quirúrgicas; cirugía general (18 %), ortopedia (16 %), cirugía pediátrica (19 %) y obstetricia (10,8 %) fueron las más frecuentes. Según la clasificación de la Sociedad Americana de Anestesiología (ASA), la mayoría de los pacientes tenían ASA 2 (39,63 %), ASA 3 (28,4 %). Se alcanzó una exposición adecuada en 11 de las 15 categorías propuestas por el ACGME y en 6 de las 15 propuestas por la Asociación Colombiana de Facultades de Medicina. Conclusiones: Se obtuvo una descripción detallada del aspecto práctico de los residentes de anestesiología durante sus tres años de formación. Esta línea de base permite ampliar el panorama a escala nacional y describir la relación con estándares internacionales.

Ebola virus VP35 interacts non-covalently with ubiquitin chains to promote viral replication.

Ebolavirus (EBOV) belongs to a family of highly pathogenic viruses that cause severe hemorrhagic fever in humans. EBOV replication requires the activity of the viral polymerase complex, which includes the cofactor and Interferon antagonist VP35. We previously showed that the covalent ubiquitination of VP35 promotes virus replication by regulating interactions with the polymerase complex. In addition, VP35 can also interact non-covalently with ubiquitin (Ub); however, the function of this interaction is unknown. Here, we report that VP35 interacts with free (unanchored) K63-linked polyUb chains. Ectopic expression of Isopeptidase T (USP5), which is known to degrade unanchored polyUb chains, reduced VP35 association with Ub and correlated with diminished polymerase activity in a minigenome assay. Using computational methods, we modeled the VP35-Ub non-covalent interacting complex, identified the VP35-Ub interacting surface, and tested mutations to validate the interface. Docking simulations identified chemical compounds that can block VP35-Ub interactions leading to reduced viral polymerase activity. Treatment with the compounds reduced replication of infectious EBOV in cells and in vivo in a mouse model. In conclusion, we identified a novel role of unanchored polyUb in regulating Ebola virus polymerase function and discovered compounds that have promising anti-Ebola virus activity.

Ebola Virus VP35 Interacts Non-Covalently with Ubiquitin Chains to Promote Viral Replication Creating New Therapeutic Opportunities.

Ebolavirus (EBOV) belongs to a family of highly pathogenic viruses that cause severe hemorrhagic fever in humans. EBOV replication requires the activity of the viral polymerase complex, which includes the co-factor and Interferon antagonist VP35. We previously showed that the covalent ubiquitination of VP35 promotes virus replication by regulating interactions with the polymerase complex. In addition, VP35 can also interact non-covalently with ubiquitin (Ub); however, the function of this interaction is unknown. Here, we report that VP35 interacts with free (unanchored) K63-linked polyUb chains. Ectopic expression of Isopeptidase T (USP5), which is known to degrade unanchored polyUb chains, reduced VP35 association with Ub and correlated with diminished polymerase activity in a minigenome assay. Using computational methods, we modeled the VP35-Ub non-covalent interacting complex, identified the VP35-Ub interacting surface and tested mutations to validate the interface. Docking simulations identified chemical compounds that can block VP35-Ub interactions leading to reduced viral polymerase activity that correlated with reduced replication of infectious EBOV. In conclusion, we identified a novel role of unanchored polyUb in regulating Ebola virus polymerase function and discovered compounds that have promising anti-Ebola virus activity.

Impact of prior dengue virus infection on Zika virus infection during pregnancy in marmosets.

Zika virus (ZIKV) infection during pregnancy causes severe developmental defects in newborns, termed congenital Zika syndrome (CZS). Factors contributing to a surge in ZIKV-associated CZS are poorly understood. One possibility is that ZIKV may exploit the antibody-dependent enhancement of infection mechanism, mediated by cross-reactive antibodies from prior dengue virus (DENV) infection, which may exacerbate ZIKV infection during pregnancy. In this study, we investigated the impact of prior DENV infection or no DENV infection on ZIKV pathogenesis during pregnancy in a total of four female common marmosets with five or six fetuses per group. The results showed that negative-sense viral RNA copies increased in the placental and fetal tissues of DENV-immune dams but not in DENV-naïve dams. In addition, viral proteins were prevalent in endothelial cells, macrophages, and neonatal Fc receptor-expressing cells in the placental trabeculae and in neuronal cells in the brains of fetuses from DENV-immune dams. DENV-immune marmosets maintained high titers of cross-reactive ZIKV-binding antibodies that were poorly neutralizing, raising the possibility that these antibodies might be involved in the exacerbation of ZIKV infection. These findings need to be verified in a larger study, and the mechanism involved in the exacerbation of ZIKV infection in DENV-immune marmosets needs further investigation. However, the results suggest a potential negative impact of preexisting DENV immunity on subsequent ZIKV infection during pregnancy in vivo.

Dengue virus NS5 degrades ERC1 during infection to antagonize NF-kB activation.

Dengue virus (DENV) is the most important human virus transmitted by mosquitos. Dengue pathogenesis is characterized by a large induction of proinflammatory cytokines. This cytokine induction varies among the four DENV serotypes (DENV1 to 4) and poses a challenge for live DENV vaccine design. Here, we identify a viral mechanism to limit NF-κB activation and cytokine secretion by the DENV protein NS5. Using proteomics, we found that NS5 binds and degrades the host protein ERC1 to antagonize NF-κB activation, limit proinflammatory cytokine secretion, and reduce cell migration. We found that ERC1 degradation involves unique properties of the methyltransferase domain of NS5 that are not conserved among the four DENV serotypes. By obtaining chimeric DENV2 and DENV4 viruses, we map the residues in NS5 for ERC1 degradation, and generate recombinant DENVs exchanging serotype properties by single amino acid substitutions. This work uncovers a function of the viral protein NS5 to limit cytokine production, critical to dengue pathogenesis. Importantly, the information provided about the serotype-specific mechanism for counteracting the antiviral response can be applied to improve live attenuated vaccines.

Validation of the TRI-SCORE in patients undergoing surgery for isolated tricuspid regurgitation.

INTRODUCTION: Estimation of peri-procedural risk in patients with tricuspid regurgitation (TR) undergoing isolated tricuspid valve surgery (ITVS) is of paramount importance. The TRI-SCORE is a new surgical risk scale specifically developed for this purpose, which ranged from 0 to 12 points and included eight parameters: right-sided heart failure signs, daily dose of furosemide ≥125 mg, glomerular filtration rate <30 mL/min, elevated bilirubin (with a value of 2 points), age ≥70 years, New York Heart Association Class III-IV, left ventricular ejection fraction <60% and moderate/severe right ventricular dysfunction (with a value of 1 point). The objective of the study was to evaluate the performance of the TRI-SCORE in an independent cohort of patients undergoing ITVS. METHODS: A retrospective observational study was performed in four centres, including consecutive adult patients undergoing ITVS for TR between 2005 and 2022. The TRI-SCORE and the traditional risk scores used in cardiac surgery (Logistic EuroScore (Log-ES) and EuroScore-II (ES-II)) were applied for each patient, and discrimination and calibration of the three scores were evaluated in the entire cohort. RESULTS: A total of 252 patients were included. The mean age was 61.5±11.2 years, 164 (65.1%) patients were female, and TR mechanism was functional in 160 (63.5%) patients. The observed in-hospital mortality was 10.3%. The estimated mortality by the Log-ES, ES-II and TRI-SCORE was 8.7±7.3%, 4.7±5.3% and 11.0±16.6%, respectively. Patients with a TRI-SCORE ≤4 and >4 had an in-hospital mortality of 1.3% and 25.0%, p=0.001, respectively. The discriminatory capacity of the TRI-SCORE had a C-statistic of 0.87 (0.81-0.92), which was significantly higher than both the Log-ES (0.65 (0.54-0.75)) and ES-II (0.67 (0.58-0.79)), p=0.001 (for both comparisons). CONCLUSION: This external validation of the TRI-SCORE demonstrated good performance to predict in-hospital mortality in patients undergoing ITVS, which was significantly better than the Log-ES and ES-II, which underestimated the observed mortality. These results support the widespread use of this score as a clinical tool.

Pathogenesis of Zika Virus Infection.

Zika virus (ZIKV) is an emerging virus from the Flaviviridae family that is transmitted to humans by mosquito vectors and represents an important health problem. Infections in pregnant women are of major concern because of potential devastating consequences during pregnancy and have been associated with microcephaly in newborns. ZIKV has a unique ability to use the host machinery to promote viral replication in a tissue-specific manner, resulting in characteristic pathological disorders. Recent studies have proposed that the host ubiquitin system acts as a major determinant of ZIKV tropism by providing the virus with an enhanced ability to enter new cells. In addition, ZIKV has developed mechanisms to evade the host immune response, thereby allowing the establishment of viral persistence and enhancing viral pathogenesis. We discuss recent reports on the mechanisms used by ZIKV to replicate efficiently, and we highlight potential new areas of research for the development of therapeutic approaches.

Análisis prospectivo y líneas estratégicas de acción para la producción agraria colombiana en un entorno pos-COVID-19 / Prospective analysis and strategic lines of action for colombian agricultural production in a pos-COVID-19 environment

El artículo desarrolla un análisis prospectivo exploratorio, orientado a la identificación de elementos de análisis, escenarios y líneas de acción estratégicas sobre la producción agraria colombiana en un entorno pos COVID-19. El método empleado desarrolló revisión de literatura, análisis estructural para la identificación de escenarios y la enunciación de líneas de acción. Los hallazgos sugieren que la crisis de demanda es el elemento más influyente, y que posibles transformaciones del sector tendrían que pasar por cambios en el uso de los territorios agrarios, en la subordinación institucional y en una novedosa perspectiva de seguridad alimentaria.

The purpose of the article was to develop a prospective exploratory analysis, oriented to the identification of elements of analysis, scenarios and strategic lines of action, on Colombian agricultural production in a post Covid-19 environment. The method used developed a literature review, structural analysis for the identification of scenarios and the enunciation of strategic lines. The findings suggest that the demand crisis is the most influential element, and that transformations in the sector would have to go through changes in the use of agrarian territories, in institutional subordination and in a novel perspective of food security.

Co-Management Reduces Mortality in Post-Sternotomy Mediastinitis.

Background: Post-sternotomy mediastinitis (PSM) is one of the most feared complications of cardiac surgery. The impact of a multidisciplinary management approach on this pathology is yet unknown. Patients and Methods: A multidisciplinary approach based on a co-management model (CMM) of care was initiated in January 2018 because of the incorporation of a hospitalist unit on a cardiac surgery department. An observational retrospective cohort study was designed to evaluate the impact of the CMM of care compared to the standard model (SM) of care in patients diagnosed with PSM. Our primary and secondary outcomes were survival time and treatment failure rate (two or more surgical procedures needed to solve PSM or PSM-related death), respectively. Data related to patient death date were collected from the Spanish National Death Index. A multivariable Cox regression model was created using those variables believed to be clinically relevant. Results: Ninety-one patients developed PSM from January 2010 to June 2020. Regarding the pre-operative clinical status, surgical procedure, and PSM severity, both groups had similar baseline characteristics. Patients were followed for a mean of 27.54 ± 30.5 months. A total of 60.3% of the SM group and 11.1% of the CMM group (p < 0.001) died. Treatment failure occurred in 53 patients (72.6%) in the SM group versus 7 (38.6%) in the CMM group (p = 0.007). The CMM independently reduced overall mortality (hazard ratio [HR], 0.11; 95% confidence interval [CI]. 0.01-0.83) and treatment failure rate (HR, 0.01; 95% CI, 0.001-0.183). Gram-positive bacterial infection (HR, 3.73; 95% CI, .6-8.3), and complete osteosynthesis material removal (HR, 0.47; 95% CI, 0.24-0.91) also influenced mortality in our model. Conclusions: A co-management care model reduced overall mortality in patients diagnosed with post-sternotomy mediastinitis.

Use of antibiotics in patients who were attacked by animals that can transmit rabies.

Animal bites have a high probability of becoming infected. In high-risk wounds, the use of antibiotics that kill the associated microorganisms is recommended. The aim of this study was to determine the use of inappropriate antibiotics in patients in different regions of Colombia who were attacked in 2020 by animals that can transmit rabies. This was a retrospective follow-up study of a cohort of patients with wounds caused by animals; the patients were affiliated with the Colombian Health System. Sociodemographic, clinical and pharmacological data were collected. A total of 280 patients were analysed. The median age was 28.0 years, and 52.1% were men. The attacks were attributed to dogs (85.0%) and cats (15.0%), and 9.6% were classified as severe exposure. Antibiotics were used in 71.1% of cases for prophylaxis and in 4.3% of the cases for the treatment of superinfection; cephalexin (37.5%), amoxicillin/clavulanic acid (15.7%) and dicloxacillin (10.4%) were the predominant antibiotics used. A total of 72.0% of patients who received antibiotics received inadequate prescriptions, and 49.2% had no indication to receive prophylaxis. Being treated in the Caribbean region (OR: 4.09; 95% CI: 1.79-9.30) and receiving analgesics (OR: 3.15; 95% CI: 1.25-7.94) were associated with a greater probability of being prescribed antibiotics inadequately; attacks resulting in severe exposure were associated with a lower probability (OR: 0.40; 95% CI: 0.16-0.98). A significant proportion of patients had no indication for prophylactic antibiotics or was prescribed antibiotics not recommended by clinical practice guidelines. There was a low prevalence of prescriptions for amoxicillin/clavulanic acid, the most recommended antibiotic.

Development and Evaluation from Laboratory to Field Trial of a Dual-Purpose Fracturing Nanofluid: Inhibition of Associated Formation Damage and Increasing Heavy Crude Oil Mobility.

This study aims to develop and evaluate fracturing nanofluids from the laboratory to the field trial with the dual purpose of increasing heavy crude oil mobility and reducing formation damage caused by the remaining fracturing fluid (FF). Two fumed silica nanoparticles of different sizes, and alumina nanoparticles were modified on the surface through basic and acidic treatments. The nanoparticles were characterized by transmission electron microscopy, dynamic light scattering, zeta potential and total acidity. The rheological behavior of the linear gel and the heavy crude oil after adding different chemical nature nanoparticles were measured at two concentrations of 100 and 1000 mg/L. Also, the contact angle assessed the alteration of the rock wettability. The nanoparticle with better performance was the raw fumed silica of 7 nm at 1000 mg/L. These were employed to prepare a fracturing nanofluid from a commercial FF. Both fluids were evaluated through their rheological behavior as a function of time at high pressure following the API RP39 test, and spontaneous imbibition tests were carried out to assess the FF's capacity to modify the wettability of the porous media. It was possible to conclude that the inclusion of 7 nm commercial silica nanoparticles allowed obtaining a reduction of 10 and 20% in the two breakers used in the commercial fracture fluid formulation without altering the rheological properties of the system. Displacement tests were also performed on proppant and rock samples at reservoir conditions of overburden and pore pressures of 3200 and 1200 psi, respectively, while the temperature was set at 77 °C and the flow rate at 0.3 cm3/min. According to the effective oil permeability, a decrease of 31% in the damage was obtained. Based on these results, the fracturing nanofluid was selected and used in the first worldwide field application in a Colombian oil field with a basic sediment and water (BSW%) of 100 and without oil production. After two weeks of the hydraulic fracture operation, crude oil was produced. Finally, one year after this work, crude oil viscosity and BSW% kept showing reductions near 75% and 33%, respectively; and having passed two years, the cumulative incremental oil production is around 120,000 barrels.

Ubiquitination of Ebola virus VP35 at lysine 309 regulates viral transcription and assembly.

Ebola virus (EBOV) VP35 is a polyfunctional protein involved in viral genome packaging, viral polymerase function, and host immune antagonism. The mechanisms regulating VP35's engagement in different functions are not well-understood. We previously showed that the host E3 ubiquitin ligase TRIM6 ubiquitinates VP35 at lysine 309 (K309) to facilitate virus replication. However, how K309 ubiquitination regulates the function of VP35 as the viral polymerase co-factor and the precise stage(s) of the EBOV replication cycle that require VP35 ubiquitination are not known. Here, we generated recombinant EBOVs encoding glycine (G) or arginine (R) mutations at VP35/K309 (rEBOV-VP35/K309G/-R) and show that both mutations prohibit VP35/K309 ubiquitination. The K309R mutant retains dsRNA binding and efficient type-I Interferon (IFN-I) antagonism due to the basic residue conservation. The rEBOV-VP35/K309G mutant loses the ability to efficiently antagonize the IFN-I response, while the rEBOV-VP35/K309R mutant's suppression is enhanced. The replication of both mutants was significantly attenuated in both IFN-competent and -deficient cells due to impaired interactions with the viral polymerase. The lack of ubiquitination on VP35/K309 or TRIM6 deficiency disrupts viral transcription with increasing severity along the transcriptional gradient. This disruption of the transcriptional gradient results in unbalanced viral protein production, including reduced synthesis of the viral transcription factor VP30. In addition, lack of ubiquitination on K309 results in enhanced interactions with the viral nucleoprotein and premature nucleocapsid packaging, leading to dysregulation of virus assembly. Overall, we identified a novel role of VP35 ubiquitination in coordinating viral transcription and assembly.

PD1 CD44 antiviral peptide as an inhibitor of the protein-protein interaction in dengue virus invasion.

Dengue virus (DENV) infection is mediated by the interaction between the virus envelope protein and cellular receptors of the host cells. In this study, we designed peptides to inhibit protein-protein interaction between dengue virus and CD44 receptor, which is one of the receptors used by DENV for entry. In silico model complexes were designed between domain III of the viral envelope protein of dengue virus 2 and the domain of human CD44 receptor using ClusPro 2.0, (https://cluspro.bu.edu/login.php), and inhibition peptides were designed with Rosetta Online-Server(http://rosie.rosettacommons.org/peptiderive). We identified one linear antiviral peptide of 18 amino acids derived from the human CD44 receptor, PD1 CD44. It did not show hemolysis or toxicity in HepG2 or BHK cell lines, nor did it stimulate the release of IL-1ß, IL-6, TNF-α, and IFN-γ, below 100 µM. It had an IC50 of 13.8 µM and maximum effective dose of 54.9 µM evaluated in BHK cells. The decrease in plaque-forming units/mL for DENV1, DENV2, DENV3, and DENV4 was 99.60%, 99.40%, 97.80%, and 70.50%, respectively, and similar results were obtained by RT-qPCR. Non-structural protein 1 release was decreased in pre- and co-treatment but not in post-treatment. Competition assays between the DN59 peptide, envelope protein, and the fragment of domain III "MDKLQLKGMSYSMCTGKF" of the viral envelope of DENV2 and PD1 CD44 showed that our peptide lost its antiviral activity. We demonstrated that our peptide decreased endosome formation, and we propose that it binds to the envelope protein of DENV, inhibiting viral invasion/fusion.

The RNA helicase DHX16 recognizes specific viral RNA to trigger RIG-I-dependent innate antiviral immunity.

Type I interferons (IFN-I) are essential to establish antiviral innate immunity. Unanchored (or free) polyubiquitin (poly-Ub) has been shown to regulate IFN-I responses. However, few unanchored poly-Ub interactors are known. To identify factors regulated by unanchored poly-Ub in a physiological setting, we developed an approach to isolate unanchored poly-Ub from lung tissue. We identified the RNA helicase DHX16 as a potential pattern recognition receptor (PRR). Silencing of DHX16 in cells and in vivo diminished IFN-I responses against influenza virus. These effects extended to members of other virus families, including Zika and SARS-CoV-2. DHX16-dependent IFN-I production requires RIG-I and unanchored K48-poly-Ub synthesized by the E3-Ub ligase TRIM6. DHX16 recognizes a signal in influenza RNA segments that undergo splicing and requires its RNA helicase motif for direct, high-affinity interactions with specific viral RNAs. Our study establishes DHX16 as a PRR that partners with RIG-I for optimal activation of antiviral immunity requiring unanchored poly-Ub.

Breastfeeding experiences during the COVID-19 pandemic in Spain:a qualitative study.

BACKGROUND: The pandemic caused by COVID-19 has affected reproductive and perinatal health both through the infection itself and, indirectly, as a consequence of changes in medical care, social policy or social and economic circumstances. The objective of this study is to explore the impact of the pandemic and of the measures adopted on breastfeeding initiation and maintenance. METHODS: A qualitative descriptive study was conducted by means in-depth semi-structured interviews, until reaching data saturation. The study was conducted between the months of January to May 2021. Participants were recruited by midwives from the Primary Care Centres of the Andalusian provinces provinces of Seville, Cádiz, Huelva, Granada, and Jaén. The interviews were conducted via phone call and were subsequently transcribed and analysed by means of reflexive inductive thematic analysis, using Braun and Clarke's thematic analysis. RESULTS: A total of 30 interviews were conducted. Five main themes and ten subthemes were developed, namely: Information received (access to the information, figure who provided the information), unequal support from the professionals during the pandemic (support to postpartum hospitalization, support received from Primary Health Care during the postpartum period), social and family support about breastfeeding (support groups, family support), impact of confinement and of social restriction measures (positive influence on breastfeeding, influence on bonding with the newborn), emotional effect of the pandemic (insecurity and fear related to contagion by coronavirus, feelings of loneliness). CONCLUSION: The use of online breastfeeding support groups through applications such as WhatsApp®, Facebook® or Instagram® has provided important breastfeeding information and support sources. The main figure identified that has provided formal breastfeeding support during this period was that of the midwife. In addition, the social restrictions inherent to the pandemic have exerted a positive effect for women in bonding and breastfeeding, as a consequence of the increase in the time spent at their homes and in the family nucleus co-living.

In Vitro Inhibition of Replication of Dengue Virus Serotypes 1-4 by siRNAs Bound to Non-Toxic Liposomes.

Dengue virus is a ssRNA+ flavivirus, which produces the dengue disease in humans. Currently, no specific treatment exists. siRNAs regulate gene expression and have been used systematically to silence viral genomes; however, they require controlled release. Liposomes show favorable results encapsulating siRNA for gene silencing. The objective herein was to design and evaluate in vitro siRNAs bound to liposomes that inhibit DENV replication. siRNAs were designed against DENV1-4 from conserved regions using siDirect2.0 and Web-BLOCK-iT™ RNAiDesigner; the initial in vitro evaluation was carried out through transfection into HepG2 cells. siRNA with silencing capacity was encapsulated in liposomes composed of D-Lin-MC3-DMA, DSPC, Chol. Cytotoxicity, hemolysis, pro-inflammatory cytokine release and antiviral activity were evaluated using plaque assay and RT-qPCR. A working concentration of siRNA was established at 40 nM. siRNA1, siRNA2, siRNA3.1, and siRNA4 were encapsulated in liposomes, and their siRNA delivery through liposomes led to a statistically significant decrease in viral titers, yielded no cytotoxicity or hemolysis and did not stimulate release of pro-inflammatory cytokines. Finally, liposomes were designed with siRNA against DENV, which proved to be safe in vitro.

The Role of the Host Ubiquitin System in Promoting Replication of Emergent Viruses.

Ubiquitination of proteins is a post-translational modification process with many different cellular functions, including protein stability, immune signaling, antiviral functions and virus replication. While ubiquitination of viral proteins can be used by the host as a defense mechanism by destroying the incoming pathogen, viruses have adapted to take advantage of this cellular process. The ubiquitin system can be hijacked by viruses to enhance various steps of the replication cycle and increase pathogenesis. Emerging viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), flaviviruses like Zika and dengue, as well as highly pathogenic viruses like Ebola and Nipah, have the ability to directly use the ubiquitination process to enhance their viral-replication cycle, and evade immune responses. Some of these mechanisms are conserved among different virus families, especially early during virus entry, providing an opportunity to develop broad-spectrum antivirals. Here, we discuss the mechanisms used by emergent viruses to exploit the host ubiquitin system, with the main focus on the role of ubiquitin in enhancing virus replication.

Trastornos electrolíticos inducidos por fármacos y sustancias tóxicas / Electrolyte disorders induced by drugs and toxic substances

Las enfermedades pueden generar un desequilibrio de electrolitos como parte de su fisiopatología, al igual que los medicamentos usados crónicamente y algunas sustancias tóxicas disponibles en nuestro medio. A pesar de todos los datos estadísticos existentes, la incidencia global de los trastornos electrolíticos secundarios a fármacos o sustancias tóxicas permanece desconocida, y, posiblemente, subregistrada; por lo tanto, el objetivo de esta revisión es analizar los trastornos electrolíticos que causan algunos medicamentos y sustancias tóxicas, y describir el mecanismo a través del cual se producen las alteraciones, en particular, del sodio, potasio, magnesio, calcio y fósforo, con el fin de alertar a los profesionales de la salud en el momento de enfrentarse a este tipo de condiciones en su práctica clínica. El conocimiento de los efectos adversos relacionados con medicamentos y tóxicos es importante para prevenir, identificar y gestionar de forma eficaz, complicaciones que son potencialmente peligrosas. Esta revisión pretende ser un referente de apoyo para los profesionales de la salud en estas situaciones

Diseases can generate an electrolyte imbalance as part of their pathophysiology, as well as chronic use of some medications, and toxic substances available in our environment. Despite all the separate statistical data, the overall incidence of fluid and electrolyte disorders secondary to drugs or toxic substances remains unknown, and possibly underreported; therefore, the objective of this review is to analyze electrolyte disorders caused by some medications and toxic substances, and describe the mechanism through which changes in sodium, potassium, magnesium, calcium and phosphorus occur, in particular, in order to alert health professionals when facing this type of conditions in their clinical practice. Knowledge of drug and toxic-related adverse effects is important to effectively prevent, identify, and manage complications that can be potentially life-threatening. This review intends to be a reference for supporting health professionals in these situations

Use of low-molecular-weight heparin in severe paraquat poisoning: a case report.

BACKGROUND: Acute paraquat ingestion remains a leading cause of mortality in developing countries. There is currently no evidence that treatment with high-dose immunosuppressants and antioxidants improves survival in patients with paraquat poisoning, and better options are urgently needed. Here, we describe the unexpected survival and recovery of a patient with a potentially fatal paraquat poisoning. CASE PRESENTATION: After ingesting 28 mL of paraquat (20% ion w/v), confirmed by a deep blue color in the urine dithionite test (UDT), a 17-year-old Hispanic Colombian boy was treated according to the hospital protocol with cyclophosphamide, methylprednisolone, N-acetylcysteine, vitamin E and propranolol. Gastrointestinal endoscopy showed extensive ulceration and necrosis. As a novelty, enoxaparin at a single dose of 60 mg was added to his treatment. Despite the evidence of severe mucosal burns in the gastrointestinal tract and high paraquat concentrations found in the UDT, the clinical condition began to improve after 1 day of treatment, with full recovery and discharge from hospital after 21 days. CONCLUSIONS: Although the amount of paraquat ingested by the patient was large and the UDT indicated severe poisoning with a somber prognosis, unexpected survival of the patient was observed, and the addition of enoxaparin was the only change from the standard treatment.