Knockdown of Dystrophin Dp71 Impairs PC12 Cells Cycle: Localization in the Spindle and Cytokinesis Structures Implies a Role for Dp71 in Cell Division

The function of dystrophin Dp71 in neuronal cells remains to be established. Previously, we revealed the involvement of this protein in both nerve growth factor (NGF)-induced neuronal differentiation and cell adhesion by isolation and characterization of PC12 neuronal cells with depleted levels of Dp71. In this work, a novel phenotype of Dp71-knockdown cells was characterized, which is their delayed growth rate. Cell cycle analyses revealed an altered behavior of Dp71-depleted cells, which consists of a delay in G0/G1 transition and an increase in apoptosis during nocodazole-induced mitotic arrest. Dp71 associates with lamin B1 and β-dystroglycan, proteins involved in aspects of the cell division cycle; therefore, we compared the distribution of Dp71 with that of lamin B1 and β-dystroglycan in PC12 cells at mitosis and cytokinesis by means of immunofluorescence and confocal microscopy analysis. All of these three proteins exhibited a similar immunostaining pattern, localized at mitotic spindle, cleavage furrow, and midbody. It is noteworthy that a drastic decreased staining in mitotic spindle, cleavage furrow, and midbody was observed for both lamin B1 and β-dystroglycan in Dp71-depleted cells. Furthermore, we demonstrated the interaction of Dp71 with lamin B1 in PC12 cells by immunoprecipitation and pull-down assays, and importantly, we revealed that knockdown of Dp71 expression caused a marked reduction in lamin B1 levels and altered localization of the nuclear envelope protein emerin. Our data indicate that Dp71 is a component of the mitotic spindle and cytokinesis multi-protein apparatuses that might modulate the cell division cycle by affecting lamin B1 and β-dystroglycan levels

Formulation and optimization of polymeric nanoparticles loaded with riolozatrione: a promising nanoformulation with potential anti-herpetic activity

Riolozatrione (RZ) is a diterpenoid compound isolated from a dichloromethane extract of the Jatropha dioica root. This compound has been shown to possess moderate anti-herpetic activity in vitro. However, because of the poor solubility of this compound in aqueous vehicles, generating a stable formulation for potential use in the treatment of infection is challenging. The aim of this work was to optimize and physiochemically characterize Eudragit® L100-55-based polymeric nanoparticles (NPs) loaded with RZ (NPR) for in vitro anti-herpetic application. The NPs formulation was initially optimized using the dichloromethane extract of J. dioica, the major component of which was RZ. The optimized NPR formulation was stable, with a size of 263 nm, polydispersity index < 0.2, the zeta potential of –37 mV, and RZ encapsulation efficiency of 89 %. The NPR showed sustained release of RZ for 48 h with release percentages of 95 and 97 % at neutral and slightly acidic pH, respectively. Regarding in vitro anti-herpetic activity, the optimized NPR showed a selectivity index for HSV-1 of ≈16 and for HSV-2 of 13

Efecto in vitro de los extractos hidroalcohólico y etanólico de semilla de papaya (Carica papaya) en Haemonchus contortus

Among the main health problems for sheep production are gastrointestinal parasites, of which the most important nematode is Haemonchus contortus. Current control strategies focus on the effectiveness, along with the low impact on the ecosystem. The study of the integral control of parasites allows the investigation of alternatives with probable anthelmintic effect, such as the use of papaya (Carica papaya) seeds. This study aimed at evaluating the in vitro effect of hydroalcoholic and ethanolic extracts of papaya seeds on eggs and larvae of H. contortus. The study was carried out with the equipment and facilities of Laboratory 3 of the Multidisciplinary Research Unit at the Facultad de Estudios Superiores Cuautitlán, UNAM. The extracts were prepared through maceration in solvent. A sheep was inoculated with 5000 L3 of H. contortus, a strain isolated and maintained in the facilities. Fecal samples were collected, and eggs were recovered for the hatch inhibition assay. The eggs and L3 were pretreated with hydroalcoholic and ethanolic extract of papaya seed (2.5 mg/ml), distilled water, ethanol 90 %, and levamisole (7.5 mg/ml). The abomasal explants technique was used to evaluate the effect of the extracts on L3 and its association to the abomasal mucosa. An ANOVA was performed for repeated samples using the Statgraphics program. The ethanolic extract of the papaya seed inhibited 92 % of hatching, and the hydroalcoholic extract 50 % (p &lt;0.05). In the case of the explants, the ethanolic extract showed 10 % of the larvae not associated with the abomasal tissue, followed by the hydroalcoholic extract with 60 % of non-associated larvae (p&lt;0.05). In conclusion, the ethanolic extract has an effect on eggs, while the hydroalcoholic extract inhibits the association to the abomasal tissue.Entre los principales problemas sanitarios para la producción ovina están los parásitos gastroentéricos; de ellos, el más importante es Haemonchus contortus. Las estrategias actuales de control enfocan la eficacia, junto con el bajo o nulo impacto al ecosistema; el estudio del control integral de parásitos permite la investigación de alternativas con probable efecto antihelmíntico, entre ellas el uso de semilla de papaya (Carica papaya). El objetivo de este trabajo fue evaluar el efecto in vitro de los extractos hidroalcohólico y etanólico de la semilla de papaya (C. papaya) sobre huevos y larvas de H. contortus. El trabajo se llevó a cabo con el equipo e instalaciones del Laboratorio 3 de la Unidad de Investigación Multidisciplinaria en la Facultad de Estudios Superiores Cuautitlán, UNAM. Se elaboraron los extractos por medio de maceración en solvente. Se inoculó un ovino con 5000 larvas de tercer estadio (L3) de H. contortus, cepa aislada y mantenida en dichas instalaciones. Se tomaron muestras de materia fecal y se recuperaron huevos para el ensayo de inhibición de la eclosión. Los huevos y las L3 fueron pretratados con extracto hidroalcohólico y etanólico de semilla de papaya (2.5 mg/ml), agua destilada, etanol 90 % y levamisol (7.5 mg/ml). Se efectuó la técnica de explantes abomasales, evaluando el efecto de los extractos sobre las L3 y su asociación a la mucosa abomasal. Se realizó un ANOVA para muestras repetidas por medio del programa Statgraphics. El extracto etanólico de semilla de papaya mostró el 92 % de inhibición de la eclosión; el extracto hidroalcohólico mostró 50 % (p&lt;0.05). En el caso de los explantes, el extracto etanólico mostró el 10 % de larvas no asociadas al tejido abomasal, seguido por el extracto hidroalcohólico, con el 60 % de larvas no asociadas (p&lt;0.05). Se concluye que el extracto etanólico tiene efecto sobre huevos, mientras que el extracto hidroalcohólico inhibe la asociación al tejido abomasal

In vitro effect of the hydroalcoholic and ethanolic extracts of papaya seeds (Carica papaya) against Haemonchus contortus

1 recurso en línea (páginas 53-59).Among the main health problems for sheep production are gastrointestinal parasites, of which the most important nematode is Haemonchus contortus. Current control strategies focus on the effectiveness, along with the low impact on the ecosystem. The study of the integral control of parasites allows the investigation of alternatives with probable anthelmintic effect, such as the use of papaya (Carica papaya) seeds. This study aimed at evaluating the in vitro effect of hydroalcoholic and ethanolic extracts of papaya seeds on eggs and larvae of H. contortus. The study was carried out with the equipment and facilities of Laboratory 3 of the Multidisciplinary Research Unit at the Facultad de Estudios Superiores Cuautitlán, UNAM. The extracts were prepared through maceration in solvent. A sheep was inoculated with 5000 L3 of H. contortus, a strain isolated and maintained in the facilities. Fecal samples were collected, and eggs were recovered for the hatch inhibition assay. The eggs and L3 were pretreated with hydroalcoholic and ethanolic extract of papaya seed (2.5 mg/ml), distilled water, ethanol 90 %, and levamisole (7.5 mg/ml). The abomasal explants technique was used to evaluate the effect of the extracts on L3 and its association to the abomasal mucosa. An ANOVA was performed for repeated samples using the Statgraphics program. The ethanolic extract of the papaya seed inhibited 92 % of hatching, and the hydroalcoholic extract 50 % (p<0.05). In the case of the explants, the ethanolic extract showed 10 % of the larvae not associated with the abomasal tissue, followed by the hydroalcoholic extract with 60 % of non-associated larvae (p<0.05). In conclusion, the ethanolic extract has an effect on eggs, while the hydroalcoholic extract inhibits the association to the abomasal tissue.Entre los principales problemas sanitarios para la producción ovina están los parásitos gastroentéricos; de ellos, el más importante es Haemonchus contortus. Las estrategias actuales de control enfocan la eficacia, junto con el bajo o nulo impacto al ecosistema; el estudio del control integral de parásitos permite la investigación de alternativas con probable efecto antihelmíntico, entre ellas el uso de semilla de papaya (Carica papaya). El objetivo de este trabajo fue evaluar el efecto in vitro de los extractos hidroalcohólico y etanólico de la semilla de papaya (C. papaya) sobre huevos y larvas de H. contortus. El trabajo se llevó a cabo con el equipo e instalaciones del Laboratorio 3 de la Unidad de Investigación Multidisciplinaria en la Facultad de Estudios Superiores Cuautitlán, UNAM. Se elaboraron los extractos por medio de maceración en solvente. Se inoculó un ovino con 5000 larvas de tercer estadio (L3) de H. contortus, cepa aislada y mantenida en dichas instalaciones. Se tomaron muestras de materia fecal y se recuperaron huevos para el ensayo de inhibición de la eclosión. Los huevos y las L3 fueron pretratados con extracto hidroalcohólico y etanólico de semilla de papaya (2.5 mg/ml), agua destilada, etanol 90 % y levamisol (7.5 mg/ml). Se efectuó la técnica de explantes abomasales, evaluando el efecto de los extractos sobre las L3 y su asociación a la mucosa abomasal. Se realizó un ANOVA para muestras repetidas por medio del programa Statgraphics. El extracto etanólico de semilla de papaya mostró el 92 % de inhibición de la eclosión; el extracto hidroalcohólico mostró 50 % (p<0.05). En el caso de los explantes, el extracto etanólico mostró el 10 % de larvas no asociadas al tejido abomasal, seguido por el extracto hidroalcohólico, con el 60 % de larvas no asociadas (p<0.05). Se concluye que el extracto etanólico tiene efecto sobre huevos, mientras que el extracto hidroalcohólico inhibe la asociación al tejido abomasal.Bibliografía: páginas 58-59

Interplay between SUMOylation and NEDDylation regulates RPL11 localization and function

The ribosomal protein L11 (RPL11) integrates different types of stress into a p53‐mediated response. Here, we analyzed the impact of the ubiquitin‐like protein SUMO on the RPL11‐mouse double‐minute 2 homolog‐p53 signaling. We show that small ubiquitin‐related modifier (SUMO)1 and SUMO2 covalently modify RPL11. We find that SUMO negatively modulates the conjugation of the ubiquitin‐like protein neural precursor cell‐expressed developmentally downregulated 8 (NEDD8) to RPL11 and promotes the translocation of the RP outside of the nucleoli. Moreover, the SUMO‐conjugating enzyme, Ubc9, is required for RPL11‐mediated activation of p53. SUMOylation of RPL11 is triggered by ribosomal stress, as well as by alternate reading frame protein upregulation. Collectively, our data identify SUMO protein conjugation to RPL11 as a new regulator of the p53‐mediated cellular response to different types of stress and reveal a previously unknown SUMO‐NEDD8 interplay

Malformations of the craniocervical junction (chiari type I and syringomyelia: classification, diagnosis and treatment)

Chiari disease (or malformation) is in general a congenital condition characterized by an anatomic defect of the base of the skull, in which the cerebellum and brain stem herniate through the foramen magnum into the cervical spinal canal. The onset of Chiari syndrome symptoms usually occurs in the second or third decade (age 25 to 45 years). Symptoms may vary between periods of exacerbation and remission. The diagnosis of Chiari type I malformation in patients with or without symptoms is established with neuroimaging techniques. The most effective therapy for patients with Chiari type I malformation/syringomyelia is surgical decompression of the foramen magnum, however there are non-surgical therapy to relieve neurophatic pain: either pharmacological and non-pharmacological. Pharmacological therapy use drugs that act on different components of pain. Non-pharmacological therapies are primarly based on spinal or peripheral electrical stimulation

Validity, reliability, and responsiveness of daily monitoring visual analog scales in MASK‐air®

Background: MASK-air® is an app that supports allergic rhinitis patients in disease control. Users register daily allergy symptoms and their impact on activities using visual analog scales (VASs). We aimed to assess the concurrent validity, reliability, and responsiveness of these daily VASs. Methods: Daily monitoring VAS data were assessed in MASK-air® users with allergic rhinitis. Concurrent validity was assessed by correlating daily VAS values with those of the EuroQol-5 Dimensions (EQ-5D) VAS, the Control of Allergic Rhinitis and Asthma Test (CARAT) score, and the Work Productivity and Activity Impairment Allergic Specific (WPAI-AS) Questionnaire (work and activity impairment scores). Intra-rater reliability was assessed in users providing multiple daily VASs within the same day. Test–retest reliability was tested in clinically stable users, as defined by the EQ-5D VAS, CARAT, or “VAS Work” (i.e., VAS assessing the impact of allergy on work). Responsiveness was determined in users with two consecutive measurements of EQ-5D-VAS or “VAS Work” indicating clinical change. Results: A total of 17,780 MASK-air® users, with 317,176 VAS days, were assessed. Concurrent validity was moderate–high (Spearman correlation coefficient range: 0.437–0.716). Intra-rater reliability intraclass correlation coefficients (ICCs) ranged between 0.870 (VAS assessing global allergy symptoms) and 0.937 (VAS assessing allergy symptoms on sleep). Test–retest reliability ICCs ranged between 0.604 and 0.878—“VAS Work” and “VAS asthma” presented the highest ICCs. Moderate/large responsiveness effect sizes were observed—the sleep VAS was associated with lower responsiveness, while the global allergy symptoms VAS demonstrated higher responsiveness. Conclusion: In MASK-air®, daily monitoring VASs have high intra-rater reliability and moderate–high validity, reliability, and responsiveness, pointing to a reliable measure of symptom loads

VER USAL: Plataforma web para el diseño y ejecución de Escape Rooms Virtuales educacionales destinados al aprendizaje de conocimientos aplicando técnicas de gamificación

Memoria ID2022-025. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2022-2023