Cell mediated immune response elicited in mice after immunization with the P64k meningococcal protein: epitope mapping

The P64k protein of Neisseria meningitidis has been reported as an immunological carrier for weak immunogens. This investigation was aimed at characterizing the T-cell response produced in primed mice and at identifying T helper cell epitopes within this molecule. BALB/c mice subcutaneously immunized with the recombinant antigen provided inguinal lymph node cells (LNC) that proliferated in the presence of P64k in a dose-dependent manner. Proliferating cells secreted IL-4 while the concentration of IL-12 remained unaltered in the culture supernatant. By testing a panel of 59 overlapping synthetic peptides spanning the entire sequence of the antigen a T-cell determinant was localized. Prime-boost and lymphoproliferation experiments, conducted with highly purified synthetic peptides, confirmed that the segment including amino acids 470-485 comprises a T-cell epitope within the P64k molecule

Sistema de talleres educativos para la vinculación escuela-familia, en el quinto año de EGB

The effective connection between schools and families is a crucial aspect of the educational success of students. This study focused on designing and evaluating educational workshops to strengthen this link in the General Basic Education School Rotary International in Canton Mocache. The design of the workshop system was based on a thorough review of the literature on school-family bonding, as well as on the specific needs identified in the educational community. Workshops were developed that addressed topics such as effective communication, parental involvement in school activities, and strategies to support learning at home. These workshops were designed taking into account cultural diversity and variations in family expectations and values. The mixed methodology employed included exploratory surveys to teachers, as well as pre-test and post-test to 35 families and 37 students respectively, along with a validation by expert criteria. Results revealed significant improvements in school-family communication, parental involvement in school activities, and student academic achievement. These findings suggest that the designed educational workshops have a positive impact on strengthening the school-family bond, thus contributing to improve the educational quality and academic development of students.La conexión efectiva entre la escuela y las familias es un aspecto crucial para el éxito educativo de los estudiantes. Este estudio se centró en diseñar y evaluar talleres educativos para fortalecer este vínculo en la escuela de educación general básica Rotary Internacional en el Cantón Mocache. El diseño del sistema de talleres se basó en una revisión exhaustiva de la literatura sobre la vinculación escuela-familia, así como en las necesidades específicas identificadas en la comunidad educativa. Se desarrollaron talleres que abordaban temas como la comunicación efectiva, la participación de los padres en las actividades escolares y estrategias para apoyar el aprendizaje en el hogar. Estos talleres fueron diseñados teniendo en cuenta la diversidad cultural y las variaciones en las expectativas y valores de las familias. La metodología mixta empleada incluyó encuestas exploratorias a docentes, así como pretest y postest a 35 familias y 37 estudiantes respectivamente, junto con una validación por criterio de expertos. Los resultados revelaron mejoras significativas en la comunicación entre la escuela y las familias, la participación de los padres en las actividades escolares, y el rendimiento académico de los estudiantes. Estos hallazgos sugieren que los talleres educativos diseñados tienen un impacto positivo en el fortalecimiento del vínculo entre la escuela y las familias, contribuyendo así a mejorar la calidad educativa y el desarrollo académico de los estudiantes

Conocimientos y aplicabilidad del código de ética en odontólogos de Cartagena, Bolívar

Tesis (Odontólogo). -- Universidad de Cartagena. Facultad de Odontología. Departamento de Investigación, 2019Dentro de esta investigación, se busca evaluar el nivel de conocimiento que tienen los Odontólogos que ejercen en la ciudad de Cartagena, Bolívar, con respecto a la ley 35 de 198

Scaling-up fermentation of Escherichia coli for production of recombinant P64k protein from Neisseria meningitidis

Background: P64k is a Neisseria meningitidis high molecular weight protein present in meningococcal vaccine preparations. The lpdA gene, which encodes for this protein, was cloned in Escherichia coli and the P64k recombinant protein was expressed in E. coli K12 GC366 cells under the control of a tryptophan promoter. P64k was expressed as an intracellular soluble protein about 28% of the total cellular protein. Several scale-up criteria of fermentation processes were studied to obtain the recombinant P64k protein at the pilot production scale. Results: The best operational conditions at a larger scale production of P64k recombinant protein were studied and compared using the four following criteria: Constant Reynold's number (Re constant), Constant impeller tip speed (n di constant), Constant power consumption per unit liquid volume (P/V constant) and Constant volumetric oxygen transfer coefficients (KLa/k constant). The highest production of the recombinant protein was achieved based on the constant KLa/k scale-up fermentation criterion, calculating the aeration rate (Q) and the impeller agitation speed (n) by iterative process, keeping constant the KLa/k value from bench scale. The P64k protein total production at the 50 l culture scale was 546 mg l-1 in comparison with the 284 mg l-1 obtained at 1.5 l bench scale. Conclusions: The methodology described herein, for the KLa/k scale-up fermentation criterion, allowed us to obtain the P64k protein at 50 l scale. A fermentation process for the production of P64k protein from N. meningitidis was established, a protein to be used in future vaccine formulations in humans.How to cite: Espinosa R, García J, Narciandi E, et al. Scaling-up fermentation of Escherichia coli for production of recombinant P64k protein from Neisseria meningitidis. Electron J Biotechnol 2018;33. https://doi.org/10.1016/j.ejbt.2018.03.004. Keywords: Escherichia coli, Fermentation process, Fermentation, Gram negative diplococcus, High molecular weight protein, Meningococcal vaccine, Neisseria meningitidis, Obligate human pathogen, Recombinant P64k protein, Scale-up, Tryptophan promote

CIGB-300 anticancer peptide regulates the protein kinase CK2-dependent phosphoproteome

Casein-kinase CK2 is a Ser/Thr protein kinase that fosters cell survival and proliferation of malignant cells. The CK2 holoenzyme, formed by the association of two catalytic alpha/alpha’ (CK2α/CK2α’) and two regulatory beta subunits (CK2β), phosphorylates diverse intracellular proteins partaking in key cellular processes. A handful of such CK2 substrates have been identified as targets for the substrate-binding anticancer peptide CIGB-300. However, since CK2β also contains a CK2 phosphorylation consensus motif, this peptide may also directly impinge on CK2 enzymatic activity, thus globally modifying the CK2-dependent phosphoproteome. To address such a possibility, firstly, we evaluated the potential interaction of CIGB-300 with CK2 subunits, both in cell-free assays and cellular lysates, as well as its effect on CK2 enzymatic activity. Then, we performed a phosphoproteomic survey focusing on early inhibitory events triggered by CIGB-300 and identified those CK2 substrates significantly inhibited along with disturbed cellular processes. Altogether, we provided here the first evidence for a direct impairment of CK2 enzymatic activity by CIGB-300. Of note, both CK2-mediated inhibitory mechanisms of this anticancer peptide (i.e., substrate- and enzyme-binding mechanism) may run in parallel in tumor cells and help to explain the different anti-neoplastic effects exerted by CIGB-300 in preclinical cancer models.Fil: Perera, Yasser. China-Cuba Biotechnology Joint Innovation Center; China. Centro de Ingeniería Genética y Biotecnología; CubaFil: Ramos, Yassel. Centro de Ingeniería Genética y Biotecnología; CubaFil: Padrón, Gabriel. Centro de Ingeniería Genética y Biotecnología; CubaFil: Caballero, Evelin. Centro de Ingeniería Genética y Biotecnología; CubaFil: Guirola, Osmany. Centro de Ingeniería Genética y Biotecnología; CubaFil: Caligiuri, Lorena Gisel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; ArgentinaFil: Lorenzo Pérez, Norailys. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; ArgentinaFil: Gottardo, María Florencia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Farina, Hernán Gabriel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Filhol, Odile. Universite Grenoble Alpes; FranciaFil: Cochet, Claude. Universite Grenoble Alpes; FranciaFil: Perea, Silvio E.. Centro de Ingeniería Genética y Biotecnología; Cub

CIGB-300 Anticancer Peptide Differentially Interacts with CK2 Subunits and Regulates Specific Signaling Mediators in a Highly Sensitive Large Cell Lung Carcinoma Cell Model

Large cell lung carcinoma (LCLC) is one form of NSCLC that spreads more aggressively than some other forms, and it represents an unmet medical need. Here, we investigated for the first time the effect of the anti-CK2 CIGB-300 peptide in NCI-H460 cells as an LCLC model. NCI-H460 cells were highly sensitive toward CIGB-300 cytotoxicity, reaching a peak of apoptosis at 6 h. Moreover, CIGB-300 slightly impaired the cell cycle of NCI-H460 cells. The CIGB-300 interactomics profile revealed in more than 300 proteins that many of them participated in biological processes relevant in cancer. Interrogation of the CK2 subunits targeting by CIGB-300 indicated the higher binding of the peptide to the CK2α′ catalytic subunit by in vivo pull-down assays plus immunoblotting analysis and confocal microscopy. The down-regulation of both phosphorylation and protein levels of the ribonuclear protein S6 (RPS6) was observed 48 h post treatment. Altogether, we have found that NCI-H460 cells are the most CIGB-300-sensitive solid tumor cell line described so far, and also, the findings we provide here uncover novel features linked to CK2 targeting by the CIGB-300 anticancer peptide

Arbuscular mycorrhizal fungus and rhizobacteria affect the physiology and performance of Sulla coronaria plants subjected to salt stress by mitigation of ionic imbalance

Salt stress has become a major menace to plant growth and productivity. The main goal of this study was to investigate the effect of inoculation with the arbuscular mycorrhizal fungi (AMF; Rhizophagus intraradices) in combination or not with plant growth‐promoting rhizobacteria (PGPR; Pseudomonas sp. (Ps) and Bacillus subtilis) on the establishment and growth of Sulla coronaria plants under saline conditions. Pot experiments were conducted in a greenhouse and S. coronaria seedlings were stressed with NaCl (100 mM) for 4 weeks. Plant biomass, mineral nutrition of shoots and activities of rhizosphere soil enzymes were assessed. Salt stress significantly reduced plant growth while increasing sodium accumulation and electrolyte leakage from leaves. However, inoculation with AMF, whether alone or combined with the PGPR Pseudomonas sp. alleviated the salt‐induced reduction of dry weight. Inoculation with only AMF increased shoot nutrient concentrations resulting in higher K+: Na+, Ca2+: Na+, and Ca2+: Mg2+ ratios compared to the non‐inoculated plants under saline conditions. The co‐inoculation with AMF and Pseudomonas sp. under saline conditions lowered shoot sodium accumulation, electrolyte leakage and malondialdehyde (MDA) levels compared to non‐inoculated plants and plants inoculated only with AMF. The findings strongly suggest that inoculation with AMF alone or co‐inoculation with AMF and Pseudomonas sp. can alleviate salt stress of plants likely through mitigation of NaCl‐induced ionic imbalance, thereby improving the nutrient profile.This work was carried out in the framework of Project AGL 2009-12530-C02-02 from Ministerio de Economía y Competitividad (Spain) and supported by the Tunisian Ministry of Higher Education and Scientific Research (LR10CBBC02), for a stay at the Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín, CSIC, Granada, Spain.Peer Reviewe