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.
Introducción: La antibiótico-resistencia es un fenómeno por el cual las bacterias logran sobrevivir al tratamiento con antimicrobianos; con incidencia en ambientes intra y extrahospitalarios como: fuentes hídricas, sector agrario/ganadero y fómites. Objetivo: Describir bacterias presentes en fómites de alta circulación en una región centro-occidental de Colombia junto a su perfil de sensibilidad fenotípica y presencia de genes para betalactamasas tipo TEM-full, OXA-3 y SHV-full. Metodología: Se aislaron cepas bacterianas de billetes, pasamanos de escaleras eléctricas y botones de cajeros automáticos; se evaluó su perfil de sensibilidad fenotípica por medio de concentración mínima inhibitoria-técnica automatizada/Vitek2® y genes para betalactamasas tipo TEM-full, OXA-3 y SHV-full mediante PCR convencional. Resultados: Se obtuvo 30 aislados; Acinetobacter baumannii complex, fue la más común; el fómite con mayor aislados y resistencia fueron los billetes; el 53% portó al menos uno de los genes estudiados. Se identificaron bacterias gramnegativas con resistencia frente a: Imipinem, Piperacilina/Tazobactam, Colistina, Ceftazidima, Tigeciclina y Ceftriaxona; bacterias grampositivas con resistencia frente a: Quinupristina/Dalfopristina, Minociclina, Tetraciclina, Teicoplanina, Nitrofuratoina, Oxacilina, Clindamicina, Trimetropina-sulfametoxazol, y Minociclina. Conclusión: Teniendo en cuenta la circulación de cepas con estas resistencias, es importante la educación en la comunidad para evitar la adquisición o propagación de infecciones por manipulación inadecuada de fómites.
Infection through the Hepatitis C virus does not have a vaccine and treatment with pegylated interferon and ribavirin can fail; which is why it may cause chronic infection and, consequently, could develop liver failure or hepatocellular carcinoma. It has been described that virus-cell recognition occurs between the E2 viral envelope protein and diverse cell receptors, with this interaction being critical in viral infection. which is why the study sought to identify inhibitory peptides of the interaction between viral E2 protein and the CD81 and CD209 receptors. Methodology: Through the RCSB protein database, crystals from the CD81 and CD209 receptors were selected, CD81/E2-HCV, CD209/E2-HCV complexes were carried out by SWISS-MODEL to generate inhibitory peptides of protein interaction through the Rosetta web server, this interaction was validated through ClusPro and finally, determined the theoretical physicochemical and cytotoxic properties of these peptides. Results: two peptides were obtained, without predicted toxicity, with a theoretical capacity of blocking the protein interaction between the E2 protein of the virus and CD81 and CD209.
Análisis del uso de antibióticos en antibiogramas de urocultivos realizados por un laboratorio clínico de la región centro-occidental de ColombiaAnalysis of the use of antibiotics in antibiograms of urine carried out by a clinical laboratory of the central-western region of ColombiaCarlos Rodríguez-Salazar 1,2* orcid.org/0000-0002-0071-1289Delia Recalde-Reyes 1,2 orcid.org/0000-0002-7752-1221Leonardo Padilla-Sanabria 1,2 orcid.org/0000-0001-5811-2791 Rodríguez-Salazar C, et al. Univ. Salud. 19(3):378-387, 2017 (Sep -Dic) [379]aeruginosa showed greater resistance and the antibiotic with the highest resistance was nalidixic acid (66.7%).
Conclusion:The study showed that there is a problem in managing, reporting and interpreting antibiograms against naturally resistant microorganisms, which could favor the development of multidrug in sensitive microorganisms of bacterial flora. A review of national and international bibliography showed similar reports; however, no author mentions intrinsic resistances, so the data of antibiotic resistance would be over evaluated.
Background:
The recent outbreak caused by SARS-CoV-2, known as COVID-19, has been cataloged as a global
catastrophe due to the growing number of infected cases and deaths since November 2019; this infectious contagious
disease, to date, does not have a vaccine available or specific treatment, which is why the number of cases continues
increasing. SARS-CoV-2 infects humans as a result of the interaction between the receptor-binding domain of the viral
spike protein and the receptor of the angiotensin-converting enzyme-2 (rACE2), located predominantly in the alveolar cells.
Objective:
To identify through computational tools, inhibitory peptides of the protein-protein interaction between the
Receptor-binding-domain of the Spike protein of SARS-CoV-2 and the angiotensin-converting enzyme-2 receptor.
Methods:
through the Research Collaboratory for Structural Bioinformatics protein database, crystals were selected and
interaction models were carried out between the viral protein and the ACE2; thereafter, the study sought and designed
inhibitory peptides of the interaction through the Rosetta web server, validated their interaction through ClusPro and,
finally, determined the theoretical physicochemical and cytotoxic properties.
Results:
A protein complex was generated modeled through ClusPro; the balanced model was selected with the lowest
binding energy. From the protein interactions of each of the crystals and from the model eight peptides of 20 residues were
obtained. The theoretical evaluation showed non-toxic peptides, six soluble in water, and two insoluble.
Conclusion:
We found eight peptides interacted with the receptor-binding-domain of the Spike Protein of SARS-CoV-2,
which could avoid contact with the cell receptor and generate interference in the infection process.
Este método de cultivo celular es una adaptación para cultivo de líneas celulares eucariotas que tradicionalmente necesitan de una atmósfera de CO2 al 5% para su crecimiento; la presente adaptación se realiza con el fin de facilitar el crecimiento de líneas celulares eucariotas en laboratorios que presenten dificultad de acceso a CO2. Este método se basa en el protocolo descrito en 2005 por el Departamento de Virología, Centro Colaborador de la OPS / OMS para Enfermedades Virales, Instituto de Medicina Tropical "Pedro Kourí", La Habana, Cuba: "Improved Dengue Virus Plaque Formation on BHK21 and LLCMK2 Cells: Evaluation of Some Factors" Mayling Alvarez, Rosmari Rodriguez-Roche, Lídice Bernardo, Luis Morier and Maria G. Guzman! Department of Virology, PAHO/WHO Collaborating Center for Viral Diseases, "Pedro Kourí" Tropical Medicine Institute, Havana, Cuba. Este protocolo ha sido probado con éxito en las líneas celulares: HepG2, Huh-7, BHK, VERO, para ensayos de crecimiento/mantenimiento, congelación y ensayos de plaqueo viral. Este protocolo fue desarrollado gracias al apoyo administrativo de la CUE Alexander von Humboldt de Armenia en convenio con la Universidad del Quindío y desarrollado dentro de la ejecución de los proyectos: Desarrollo de una prueba de western blot para detección de antígenos virales de dengue serotipos 1–4 y Evaluación de la citotoxicidad y producción de citoquinas pro-inflamatorias (IL-1b, INF-y, IL-6, IL-10, TNF α) de leucocitos humanos que interactúan con liposomas cargados con siRNA contra el virus del dengue.
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