Chimerization of antibodies by isolation of rearranged genomic variable regions by the polymerase chain reaction

We describe a new method for amplification, by polymerase chain reaction (PCR), of rearranged segments encoding the variable part of light and heavy chains of an antibody (Ab) from the chromosomal DNA of hybridoma cells for the chimerization ofAbs. A fundamental prerequisite for this is the knowledge ofthe exact sequences in the 5’-untranslated region of light and heavy chain mRNA, and of the joining segment used for rearrangement. This allows the design of nondegenerated oligodeoxyribonucleotides for PCR. The primer design permits directional cloning of the amplified, promoterless fragments into cassette vectors, in which they will be linked to the appropriate human constant domains and immunoglobulin (Ig) promoter/enhancer elements. The method is illustrated for chimerization of an Ab directed against the human T-lymphocyte antigen, CD4. The chimerized Ab is secreted in abundant quantities after transfection of the engineered plasmids into non-Ig-producing myeloma cells

Comunicación humanitaria para el desarrollo, comunicación social: entre ética y estética. Propuestas teóricas de definición

Desde hace décadas, el desarrollo es un espacio de intervención y práctica sociopolítica y cultural en cuyo proceso de institucionalización, legitimación y transformación ha incidido la práctica de investigación y la reflexión teórica de ámbitos muy dispares, entre ellos, el de comunicación. En la producción teórica y de investigación – así como en las prácticas de acción social- encontramos conceptos de fronteras difusas como: comunicación para el desarrollo, comunicación del desarrollo, comunicación humanitaria, información humanitaria, etc. A partir del análisis de dos distintos tipos de comunicación (la de las ONGD españolas y de las instituciones públicas para el desarrollo), este artículo propone una reflexión sobre la necesidad de delimitar un espacio teórico desde el que reflexionar sobre prácticas comunicativas que atañen a representaciones sobre el sufrimiento humano –en cuanto un sufrimiento evitable conectado con las condiciones socio-culturales, económicas y políticas-, prácticas que conllevan una dimensión ético-política inexorable y que, por tanto, nos sitúan ante prácticas comunicativas que no puede responder a criterios puramente estéticos aún cuando primen en ellas criterios funcionales de distinto orden.For decades, development has been an area of intervention and sociopolitical practice and culture in which the process of institutionalization, legitimation, and transformation has affected the practice of research and theoretical reflection of very disparate fields, including the media. In the theoretical and research - as well as social action practices, we find concepts of fuzzy boundaries as development communication, development communication, humanitarian information, etc. From the analysis of two different types of communication (the Spanish NGOs and public institutions for development), this article proposes a reflection on the need to define a theoretical space from which to reflect on communication practices regarding representations on the human suffering in avoidable suffering connected with the sociocultural, economic and political-practices involving ethical-political dimension inexorable and therefore put us in communicative practices can not answer purely to esthetic criteria

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Treball presentat a l'assignatura de Deontologia i Veterinària Legal (21223

SUCCULENT PLANT SPECIES FROM THE CRASSULACEAE FAMILY PRESENT IN THE GREENHOUSES OF THE BOTANICAL GARDEN "AL. BUIA" FROM CRAIOVA

Dissemination of information regarding the existing plant collections in the Botanical Garden "Al. Buia" from Craiova is a necessity for knowing, understanding and appreciating its plant heritage at its fair value. Through the collections it holds, it can be said that the Botanical Garden effectively contributes to the conservation of plant diversity, as some rare and threatened species from Europe and other regions of the world are also present here. Also, an interesting group of plants grown in the greenhouses of the Craiova Botanical Garden is represented by succulent plants, with representatives from several families, of which species belonging to the Crassulaceae family are presented in this work. This family includes mostly herbaceous species, rarely frutescent plants with succulent leaves that grow spontaneously in dry, rocky, water-poor places in the warm and temperate regions of the globe. The Crassulaceae family is well represented in the greenhouse by species of the genera Adromischus, Aeonium, Aichryson, Bryophyllum, Crassula, Echeveria, Kalanchoe, Monanthes, Pachyphytum, Sedum, Sinocrassula, generally originating in the arid areas of South Africa. Some species are native to the Canary Islands, South America, Central America and Mexico, but all exhibit adaptations to xerophytism, such as covering with trichomes or a wax coating

Analysis of different structural steelwork solutions for an industrial building

[Resumen] En este proyecto se ha realizado el diseño, cálculo y optimización de la estructura metálica de una nave industrial. Además del cálculo de una serie de variantes de la misma. Se trata de una estructura biempotrada constituida por 8 pórticos formados por perfiles IPE y HEA. Su luz estructural será de 22 metros con separación entre pórticos de 6 metros alcanzando así una longitud total de 42 metros. En lo que respecta a la altura de pilares, será de 7 metros en la parte exterior y de 8,1 metros en coronación.[Resumo] Neste proxecto realizouse o deseño, cálculo e optimización da estrutura metálica dunha nave industrial. Ademais do cálculo dunha serie de variantes da mesma. Trátase dunha estrutura biempotrada constituida por 8 pórticos fomados por perfís IPE e HEA. A luz estrutural será de 22 metros con separación entre pórticos de 6 metros acadando así unha lonxitude total de 42 metros. No que respecta a á altura de piares, será de 7 metros na parte exterior e de 8,1 metros en coroación.[Abstract] In this project, the design, calculation and optimisation of the metallic structure of a warehouse has been carried out. In addition, a series of variants of the same will be calculated. It is a biembedded structure composed of 8 porticoes formed by IPE and HEA profiles. Its structural light will be 22 meters with separation between porticoes of 6 meters thus reaching a total length of 42 meters. Regarding the height of the pillars, it will be 7 meters on the outside and 8,1 meters in coronation.Traballo fin de grao (UDC.EPS). Enxeñaría en tecnoloxías industriais. Curso 2021/202

Combinatorial functions of two chimeric antibodies directed to human CD4 and one directed to the a-chain of the human interleukin-2 receptor

The general feasibility of chimerization of monoclonal antibodies (mAbs) has already been shown for a large number of them. In order to evaluate in vitro parameters relevant to immunosuppressive therapy, we have chimerized and synthesized two anti-CD4 mAbs recognizing two different epitopes on the human T-lymphocyte antigen, CD4. The chimerized mAbs are produced at levels corresponding to those of the original hybridoma cell lines. With respect to activation of human complement, the individual Abs are negative; however, when used in combination, complement activation was performed. When applied in combination, they were found to modulate the CD4 antigen, whereas the individual mAb do not display this property. Individually they mediate an up to 60% inhibition of the mixed lymphocyte reaction (MLR). However, by combination of an anti-CD4 mAb with one directed against the a-chain of the human IL2 receptor, nearly 100% inhibition of the MLR was achieved, even with reduced dosage of the mAbs. Our data suggest that the combination of an anti-CD4 mAb and an anti-IL2Rcc chain mAb is more effective with respect to immunosuppression than each mAb by itself, indicating that this mAb cocktail could be a new strategy for immunosuppressive therapy

Microglial responses in the human Alzheimer’s disease frontal cortex

The continuing failure to develop an effective treatment for Alzheimer’s disease (AD) reveals the complexity for this pathology. Increasing evidence indicates that neuroinflammation involving particularly microglial cells contributes to AD pathogenesis. The actual view, based on the findings in APP based models, gives a cytotoxic/proinflammatory role to activated microglia. However, we have previously reported a limited activation and microglial degeneration in the hippocampus of AD patients in contrast with that observed in amyloidogenic models. Here, we evaluated the microglial response in a different region of AD brains, the frontal cortex. Post mortem tissue from controls (Braak 0-II) and AD patients (Braak V-VI) including familial cases, were obtained from Spain Neurological Tissue Banks. Cellular (immunohistochemistry and image analysis) and molecular (qPCR and western blots) approaches were performed. Frontal cortex of AD patients (Braak V-VI) showed strong microglial activation similar to that observed in amyloidogenic mice. These strongly activated microglial cells, predominantly located surrounding amyloid plaques, could drive the AD pathology and, in consequence, could be implicated in the pathology progression. Furthermore, different microglial responses were observed between sporadic and familial AD cases. These findings in the frontal cortex were highly in contrast to the attenuated activation and degenerative morphology displayed by microglial cells in the hippocampus of AD patients. Regional differences in the microglial response suggest different functional states of microglial cells in a region-specific manner. All together, these data provide a better understanding of the immunological mechanisms underlying AD progression and uncover new potential therapeutic targets to fight this devastating neurodegenerative disease.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Supported by PI18/01557 (AG) and PI18/01556 (JV) grants from ISCiii of Spain co-financed by FEDER funds from European Unio

A Tobacco Homolog of DCN1 is Involved in Cellular Reprogramming and in Developmental Transitions

Plant proteomes show remarkable plasticity in reaction to environmental challenges and during developmental transitions. Some of this adaptability comes from ubiquitin-mediated protein destruction regulated by cullin-RING E3 ubiquitin ligases (CRLs). CRLs are activated through modification of the cullin subunit with the ubiquitin-like protein RUB/NEDD8 by an E3 ligase called defective in cullin neddylation 1 (DCN1). Here we show that tobacco DCN1 binds ubiquitin and RUB/NEDD8, and associates with cullin. When knocked down by RNAi, tobacco pollen formation stopped and zygotic embryogenesis was blocked around the globular stage. Additionally, we found that RNAi of DCN1 inhibited the stress-triggered reprogramming of cultured microspores from their intrinsic gametophytic mode of development to an embryogenic state. This stress-induced developmental switch is a known feature in many important crops and leads ultimately to the formation of haploid embryos and plants. Compensating the RNAi effect by re-transformation with a promoter-silencing construct restored pollen development and zygotic embryogenesis, as well as the ability for stress-induced formation of embryogenic microspores. Overexpression of DCN1, however, accelerated pollen tube growth and increased the potential for microspore reprogramming. These results demonstrate that the biochemical function of DCN1 is conserved in plants and that its activity is specifically required for transitions during pollen development and embryogenesis, and for pollen tube tip growth

Chimerization of antibodies by isolation of rearranged genomic variable regions by the polymerase chain reaction

We describe a new method for amplification, by polymerase chain reaction (PCR), of rearranged segments encoding the variable part of light and heavy chains of an antibody (Ab) from the chromosomal DNA of hybridoma cells for the chimerization ofAbs. A fundamental prerequisite for this is the knowledge ofthe exact sequences in the 5’-untranslated region of light and heavy chain mRNA, and of the joining segment used for rearrangement. This allows the design of nondegenerated oligodeoxyribonucleotides for PCR. The primer design permits directional cloning of the amplified, promoterless fragments into cassette vectors, in which they will be linked to the appropriate human constant domains and immunoglobulin (Ig) promoter/enhancer elements. The method is illustrated for chimerization of an Ab directed against the human T-lymphocyte antigen, CD4. The chimerized Ab is secreted in abundant quantities after transfection of the engineered plasmids into non-Ig-producing myeloma cells