La vacunación en el contexto internacional

La vacunación es uno de los avances con mayor impacto en la población respecto a la Salud Pública. La vacunación es un sistema de inmunización del organismo frente a una determinada enfermedad infectocontagiosa. A lo largo de la historia, los diferentes avances científicos han permitido mejorar el acto de la vacunación. El objetivo de este trabajo es comparar la vacunación en España con el contexto europeo de su entorno y conocer la vacunación administrada en países africanos y contrastar con contexto europeo. Se ha realizado una revisión bibliográfica sistemática a través de las bases de datos más recomendadas (Pubmed, Dialnet..), páginas institucionales y documentos oficiales. Desde el inicio de la vacunación con Edward Jenner, hasta la actualidad ha habido diversos cambios, pero sigue siendo un tema de gran interés mundial. En primer lugar, España posee un calendario vacunal común, adoptando cada comunidad autónoma sus propias normas y modificaciones. En lo que refiere al continente africano, se administran algunas menos vacunas que en la Unión Europea, por el hecho de ser una región en vías de desarrollo, con más enfermedades contagio-infecciosas y con una menor posesión de recursos que otros territorios continentales. Las vacunaciones administradas en las diferentes comunidades autónomas españolas son muy semejantes, pero al compararlas con los países de la Unión Europea, difieran algo más, optando algunos países por la obligatoriedad de la vacunación, tras el resurgimiento de nuevas epidemias y tras la aparición del movimiento anti-vacuna. África difiere aún más, ya que únicamente se administran las mínimas vacunas recomendadas por la OMS.Grado en Enfermerí

Geminivirus C2 protein represses genes involved in sulphur assimilation and this effect can be counteracted by jasmonate treatment

Geminiviruses are plant viruses that infect a broad range of crops and cause extensive losses worldwide, having an important economic impact. C2, a multifunctional pathogenicity factor encoded by geminiviruses, has been recently shown to suppress the responses to jasmonates in the host plant, which might at least partially explain its well-established role in pathogenicity. Sulphur is one of the essential macro-elements for plant life, and is considered to have a role in plant defence, in a phenomenon named sulphur-induced resistance (SIR) or sulphur-enhanced defence (SED). In this work, we show that geminivirus C2 protein represses the expression of genes involved in the sulphur assimilation pathway in Arabidopsis, but, interestingly, this effect can be neutralized by exogenous jasmonate treatment. These preliminary results may raise the idea that geminiviruses might be affecting sulphur metabolism, and maybe counteracting SIR/SED, through the manipulation of the jasmonate signalling pathway, which would define a novel strategy in plant-virus interactions and may unveil SIR/SED as an important player in the plant defence against viruses.Ministerio de Ciencia y Innovación/FEDER AGL2007-66062-C02-02/AGR AGL2010-22287-CO2European Regional Development Fund (ERDF) BIO2010-15201Junta de Andalucía BIO­27

Quality Assessment of Virus-Like Particles at Single Particle Level : A Comparative Study

Virus-like particles (VLPs) have emerged as a powerful scaffold for antigen presentation and delivery strategies. Compared to single protein-based therapeutics, quality assessment requires a higher degree of refinement due to the structure of VLPs and their similar properties to extracellular vesicles (EVs). Advances in the field of nanotechnology with single particle and high-resolution analysis techniques provide appealing approaches to VLP characterization. In this study, six different biophysical methods have been assessed for the characterization of HIV-1-based VLPs produced in mammalian and insect cell platforms. Sample preparation and equipment set-up were optimized for the six strategies evaluated. Electron Microscopy (EM) disclosed the presence of several types of EVs within VLP preparations and cryogenic transmission electron microscopy (cryo-TEM) resulted in the best technique to resolve the VLP ultrastructure. The use of super-resolution fluorescence microscopy (SRFM), nanoparticle tracking analysis (NTA) and flow virometry enabled the high throughput quantification of VLPs. Interestingly, differences in the determination of nanoparticle concentration were observed between techniques. Moreover, NTA and flow virometry allowed the quantification of both EVs and VLPs within the same experiment while analyzing particle size distribution (PSD), simultaneously. These results provide new insights into the use of different analytical tools to monitor the production of nanoparticle-based biologicals and their associated contaminants

Development of a purification process for HIV-1 VLPs, from supernatant to lyophilization

HIV-1-based virus-like particles (VLPs) have high potential as scaffold for the development of chimeric or multivalent vaccines by functionalizing them with specific antigens [1], [2]. The obtention of vaccine formulations independent of cold chain is desirable to facilitate transportation and administration worldwide [3]. Recently, efforts are being made to develop cost-effective scalable processes to obtain these particles. The present study aims to compare some of the most used downstream processing (DSP) technologies for capture and purification of VLPs, taking especial consideration in using technologies enabling operation at large scale [4]. First, suspension adapted HEK 293 cells cultured in chemically defined cell culture media were used to produce the Gag-eGFP VLPs. Then, some steps of the purification process were studied, including a primary and secondary clarification by depth filtration and filtration respectively, an intermediate step by tangential flow filtration (TFF) or multimodal chromatography (MC), a capture step by ion exchange (IEC), heparin affinity (AC) and hydrophobic interaction chromatography (HIC), a polishing step by size exclusion chromatography (SEC) and a finally lyophilization step by freeze-drying process. Different operation units were tested for each step. Finally, a complete DSP train was implemented using the best results obtained in each stage. A concentration of 2.2 ± 0.8·109 VLPs/mL in the lyophilized samples was obtained after its storage at room temperature for 2 months. The morphology and structural integrity were further assessed by cryo-TEM. These first results in enveloped VLP lyophilization offer great promise to overcome the difficulty to distribute vaccines in poorly served remote rural areas and increase vaccine stability until their administration. Likewise, the purification methodologies proposed here could be easily scaled up and applied to purify similar enveloped viruses and vesicles. Please click Additional File below for the presentation

Lipids and lipoxidation in human brain aging. Mitochondrial ATP-synthase as a key lipoxidation target

The human brain is a target of the aging process like other cell systems of the human body. Specific regions of the human brain exhibit differential vulnerabilities to the aging process. Yet the underlying mechanisms that sustain the preservation or deterioration of neurons and cerebral functions are unknown. In this review, we focus attention on the role of lipids and the importance of the cross-regionally different vulnerabilities in human brain aging. In particular, we first consider a brief approach to the lipidomics of human brain, the relationship between lipids and lipoxidative damage, the role of lipids in human brain aging, and the specific targets of lipoxidative damage in human brain and during aging. It is proposed that the restricted set of modified proteins and the functional categories involved may be considered putative collaborative factors contributing to neuronal aging, and that mitochondrial ATP synthase is a key lipoxidative target in human brain aging

Formation of stimuli-responsive cyclophanes by self-assembly: the case of carbazole-based biradicals

Dynamic covalent bonds has recently received lot of attention because of their unique feature to become reversible under mild conditions.[1] In this context, π-conjugated biradical compounds has emerged as essential building blocks.[2] For instance, we have demonstrated that 2,7-dicyanomethylene-9-(2-ethylhexyl)carbazole biradical reversibly converts to a macrocycle cyclophane upon soft stimuli (temperature, pressure, light), showing strong chromic effects.[3] We now extent this study towards longer conjugated carbazole backbone (i.e., indolocarbazole shown in Figure 1), aiming at investigating how the elongation of the conjugated backbone impacts on the formation of stimuli-responsive cyclophanes. The self-assembly process is investigated both in solution and solid state by linking theory and experiments.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

At-line multi-angle light scattering detector for faster process development in enveloped virus-like particle purification

At-line static light scattering and fluorescence monitoring allows direct in-process tracking of fluorescent virus-like particles. We have demonstrated this by coupling at-line multi-angle light scattering and fluorescence detectors to the downstream processing of enveloped virus-like particles. Since light scattering intensity is directly proportional to particle concentration, our strategy allowed a swift identification of product containing fractions and rapid process development. Virus-like particles containing the Human Immunodeficiency Virus-1 Gag protein fused to the Green Fluorescence protein were produced in Human Embryonic Kidney 293 cells by transient transfection. A single-column anion-exchange chromatography method was used for direct capture and purification. The majority of host-cell protein impurities passed through the column without binding. Virus-like particles bound to the column were eluted by linear or step salt gradients. Particles recovered in the step gradient purification were characterized by nanoparticle tracking analysis, size exclusion chromatography coupled to multi-angle light scattering and fluorescence detectors and transmission electron microscopy. A total recovery of 66% for the fluorescent particles was obtained with a 50% yield in the main product peak. Virus-like particles were concentrated 17-fold to final a concentration of 4.45 × 10 particles/mL. Simple buffers and operation make this process suitable for large scale purposes

Carbazole-based Diradicals for Dynamic Covalent Chemistry

Dynamic covalent chemistry (DCC) is focused on the creation of structural scaffolds based on chemical components that interact through strong but reversible bonds. In fact, dynamic covalent bonds receive lot of attention because of their unique feature to become reversible under mild conditions.1 conjugated diradical compounds has emerged as essential building blocks in DCC.2 In this work, we will review our most recent works on the formation of stimuli-responsive cyclophanes by self-assembly of carbazole-based diradicals. To this end, we use a combined experimental and theoretical approach that links vibrational spectroscopy with DFT calculations. In this sense, it is interesting to note that we have recently demonstrated the potential of a para-substituted carbazole with terminal dicyanomethylene (DCM) groups to act as building blocks in DCC.3 This quinoid carbazole monomer transforms to a macrocycle cyclophane upon soft external stimuli (temperature, pressure, light), showing strong chromic features. In addition, we have also recently explored how the different DCM substitution position affects the interesting chromoactive properties of carbazole compounds.4 Finally, we are currently exploring the effect of the elongation of the carbazole backbone on the formation of stimuli-responsive cyclophanes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Multiresponsive chromic soft materials: formation of strongly coupled σ-dimers from IndoloCarbazole-based biradicaloids

Dynamic covalent chemistry is focused on the creation of structural scaffolds based on chemical components that interact through strong but reversible bonds. In fact, dynamic covalent bonds receive lot of attention because of their unique feature to become reversible under mild conditions.1 π-conjugated biradical compounds has emerged as essential building blocks in DCC (dynamic covalent chemistry).2 We have recently demonstrated the potential of a para-substituted carbazole with terminal dicyanomethylene groups to act as building blocks in DCC.3 In fact, this quinoid carbazole monomer transform to a macrocycle cyclophane upon soft external stimuli (temperature, pressure, light), showing strong chromic features. Here, we explore the effect of the elongation of the carbazole backbone on the formation of stimuli-responsive cyclophanes by self-assembly. To this end, we use a join experimental and theoretical approach that links vibrational spectroscopy (Raman and IR) with DFT calculationsUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Differential glycosylation and extracellular vesicle biogenesis in HEK293 upon transient transfection

Vaccine therapies based on virus-like particles (VLPs) are currently increasing relevance due to the strong immune response they elicit and their manufacture advantages when compared to traditional biopharmaceuticals. During VLP production using mammalian cell-based platforms, different extracellular vesicles (EVs) are coproduced, leading to the need of a complex downstream purification process. Currently there is no effective and efficient method to separate VLPs from EVs which share very similar density and physicochemical properties1,2. Different methods to characterize the EV composition and their protein content and glycosylation signature are used in this work to further understand their biochemical nature. First, a sucrose cushion ultracentrifugation was carried out to isolate the VLP fraction, also containing co-purified EVs. Following, a multiplexed quantitative proteomic approach was used to characterize the VLP-copurified secretome. Three conditions were studied, a non-transfected condition, transiently transfected with an empty plasmid (mock) and with a plasmid for Gag VLP production. Please click Download on the upper right corner to see the full abstract