El contrapunto en el álbum ilustrado infantil. Un análisis multimodal para su implementación en la asignatura Ilustración Infantil del Máster de Dibujo (Universidad de Granada)

El álbum ilustrado infantil es una de las publicaciones para niños con mayor grado de experimentación literaria-plástica. La clave de estos libros radica en la relación entre la imagen y el texto, donde ambos medios colaboran entre sí para contar una historia común. Esta investigación se centra en los álbumes donde la relación multimodal es fundamentalmente de contrapunto, es decir, la imagen y el texto cuentan historias distintas, que incluso pueden llegar a contradecirse. El objetivo de esta aportación es doble: en primer lugar, se profundiza en los recursos de interacción verbal-icónica que dan lugar al contrapunto en el álbum ilustrado; y, a posteriori, se diseña una actividad didáctica enfocada en la descodificación textual y visual de los álbumes analizados, con el objeto de implementarla como parte de la programación docente del Máster de Dibujo. El método empleado se basa en un estudio cualitativo de cinco álbumes contemporáneos publicados en español, tomando como punto de partida las categorías de contrapunto de Nikolajeva y Scott, y las clasificaciones de recursos metaficcionales descritos por Lewis y Zabala. Los resultados del análisis sirven de base para el diseño de la actividad, donde los estudiantes trabajarán los conceptos explicados previamente en el aula. El análisis revela un claro predominio de los recursos denominados “estilo”, “punto de vista” y “género” para crear el contrapunto fundamental en el álbum. La actividad didáctica muestra las dificultades de identificación del contrapunto en los álbumes no contradictorios. De esta manera, para mejorar la eficacia de la actividad, se proponen algunos ajustes, como son la ampliación del número de álbumes para reducir los grupos de trabajo, y la incorporación de álbumes de complemento para facilitar la comprensión de los conceptos por comparación.

Sobre los mitos del Rey Arturo y de la magia

Los ensayistas y literatos C. S. Lewis y Charles Williams recogieron la Materia de Bretaña, con su valor mítico, y lo plasmaron dualmente a través de las leyendas artúricas y la categorización de la magia a través de la división convencional entre magia blanca y magia negra. En este último supuesto, respetaron la división propia de los mitos de las artes mágicas entre teúrgia y goetia —magia natural y magia demoníaca— en continuidad con los planteamientos de la renacentista Academia Platónica de Florencia. Esta recepción del mito aparece nítidamente en las obras de C. S. Lewis, Esa horrible fortaleza, y de Charles Williams, Guerra en el cielo. Se trata de marcos de ficción, donde realizan un ejercicio mitopoético que vincula los mitos artúricos y mágicos para responder desde un fundamento metafísico y trascendente a preguntas perennes de la condición humana.

Glycomic Characterization of Platelets from Patients with Immune Thrombocytopenia

Introduction: Platelet glycoproteins are key contributors to platelet function but their glycans structure is unclear. Alterations in glycan composition have been reported to impact platelet clearance under physiological conditions and in the disease mechanism of immune thrombocytopenia (ITP). Therefore, this study sought to characterize glycan structures in human platelets from healthy control individuals and ITP patients using mass spectrometry (MS)-based glycomics approach, andto compare their glycomic profiles to facilitate understanding of glycan alterations in ITP. Methods: Glycan residues on platelet surface were determined by flow cytometry. Platelet lysates (1×10 8 platelets) from 4 healthy controls and from 4 ITP patients with a clear anomalous glycosylation pattern were characterized by MALDI-MS based glycomic approaches. N-linked glycans were released from platelet glycoproteins by PNGase F digestion and subsequently purified with a Sep-Pak C18 reverse phase cartridge. O-linked glycans were released by reductive elimination. Both pools of glycans were permethylated prior to MALDI-TOF MS to obtain an initial carbohydrate profile. Selected glycan molecular ion species were analyzed by MALDI-TOF-TOF MS/MS before and after digesting with exoglycosidases. Results: Glycans present in platelets from healthy controls and from ITP patients were largely consistent. The MS spectra for N-glycans showed a mixture of high mannose glycans (m/z 1579.8, 1783.9, 1988.0, 2192.1 and 2396.2); complex glycans (m/z 2966.5, 3776.9 and 4587.4); and bisected or truncated glycans (m/z 3211.6 and 4022.1). The spectra showed the presence of bi-, tri- and tetra-antennary complex glycans, which varied in their level of sialylation.Figure 1 shows the relative abundance ratio within eight families of core glycan structures. Platelets from ITP patients showed a consistent increase in desialylated structures such as Hex 5HexNAc 4Fuc 1. In addition to different amounts of attached sialic acid, varying levels of fucosylation were observed; ranging from the addition of a single core Fuc (m/z 2244.1), to the addition of up to three Fuc residues (m/z 3402.7). Collision-activated decomposition (CAD) MALDI-TOF/TOF analysis was performed to generate fragment ions from molecular ions detected in MALDI-TOF profiling for detailed sequencing of platelet N-glycans. This analysis suggested the presence of three isoforms: (i) sialylated tetra-antennary structure with core fucosylation and one Lewis x/a antenna; (ii) sialylated tetra-antennary structure with one Lewis y/b antenna; (ii) sialylated, core-fucosylated tri-antennary structure with one LacNAc extension and one Lewis x/a antenna. Sialidase S digestion was used to highlight the extent of desialylation in the presence of sialidase. Noteworthy, the ratio of non-sialylated bi-and tri-antennary N-glycans in ITP patients were higher than that in controls. This enzymatic digestion confirms the presence of α2,3-linked Neu5Ac on platelet glycans. Remaining sialylated structures may possess α2,6-linked Neu5Ac. O-glycan profiles obtained showed the predominance of core 1 and core 2 structures (Figure 2). The two most dominant glycan structures in core 1 were sialyl T antigen (GalNAc 1Gal 1NeuAc 1; m/z 895.5) and disialyl T antigen (GalNAc 1Gal 1NeuAc 2; m/z 1256.7), being the latter less abundant in ITP patients. The core 2 structure was modified by the addition of fucose and/or sialic acid residues (m/z 1157.7, 1344.8, 1518.9 and 1706.0). Conclusion: N- and O-glycan structures in human platelets were characterized by MALDI-TOF MS profiling to reveal interesting structural features including the presence of sialylLewis x/a epitope, Lewis x/a epitope, Lewis y/b epitope and LacNAc extensions in complex type N-glycans. Presence of terminal sialic acid and sialylLewis x/a on platelet N-glycan antenna also suggest their potentialfunction as ligands for siglecs that are associated with cell signaling functions. Siglec-1 and -2 have been suggested to have potential roles in ethiopathogenesis of autoimmune diseases. Desialylation observed in glycans of platelets from ITP patients, might trigger immune system activation in these patients. This research was funded by ISCIII-Fondos FEDER PI19/00772 and Platelet Disorder Support Association Figure 1 Figure 1. Disclosures Butta: Roche: Speakers Bureau; Takeda: Research Funding, Speakers Bureau; CSL-Behring: Research Funding; Novo-Nordisk: Speakers Bureau. Canales: F. Hoffmann-La Roche Ltd: Consultancy, Honoraria, Speakers Bureau; Karyopharm: Consultancy, Honoraria; Takeda: Consultancy, Honoraria, Speakers Bureau; iQone: Honoraria; Sandoz: Honoraria, Speakers Bureau; Incyte: Consultancy; Janssen: Consultancy, Honoraria, Speakers Bureau; Gilead/Kite: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Sanofi: Consultancy; Eusa Pharma: Consultancy, Honoraria; Celgene/Bristol-Myers Squibb: Consultancy, Honoraria. Jiménez-Yuste: Pfizer: Consultancy, Honoraria, Research Funding; F. Hoffmann-La Roche Ltd: Consultancy, Honoraria, Research Funding; BioMarin: Consultancy; Sobi: Consultancy, Honoraria, Research Funding; NovoNordisk: Consultancy, Honoraria, Research Funding; Octapharma: Consultancy, Honoraria, Research Funding; Sanofi: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Bayer: Consultancy, Honoraria, Research Funding; CSL Behring: Consultancy, Honoraria, Research Funding; Grifols: Consultancy, Honoraria, Research Funding. Alvarez Román: Octapharma: Consultancy, Honoraria, Research Funding; Biomarin: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; Pfizer: Consultancy, Honoraria, Research Funding; Bayer: Consultancy, Honoraria, Research Funding; CSL-Behring: Consultancy, Honoraria, Research Funding; Grifols: Consultancy, Honoraria, Research Funding; Novo-Nordisk: Consultancy, Honoraria, Research Funding; Sobi: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding.

781 GT-001 - anti-Lewis Y antibody with superior fine-specificity and reduced off-target binding

BackgroundThe Lewis Y (CD174) carbohydrate antigen is widely expressed in primary and metastatic epithelial tumors like colon, lung, ovarian, and breast. Targeting Lewis Y for cancer therapy was pursued before, however, other anti-Lewis Y antibodies tested in clinical trials showed cross-reactivity to related carbohydrate structures expressed on blood cells and mostly failed for efficacy and/or safety reasons.1–4 We have developed a humanized antibody (GT-001) that shows superior fine-specificity and higher affinity compared to clinically tested anti-Lewis Y antibodies BR96 and h3S193.MethodsThe specificity and cross-reactivity of GT-001, BR96 and h3S193 were compared. Cross-reactivity binding to related carbohydrate PAA-conjugates was tested via ELISA and affinity towards Lewis Y-PAA was measured using switchSENSE® technology (DRX2, Dynamic Biosensors). Functional binding to several tumor cell lines and healthy human leukocytes was analyzed via flow cytometry. Binding of GT-001 to different cancer indications was analyzed by immunohistochemistry. Inhibition of tumor cell proliferation was tested using GT-001 coupled to ProtG-MMAE.ResultsGT-001 is strictly specific for Lewis Y and does not cross-react with >90 related carbohydrate structures tested. Our lead candidate shows superior fine-specificity compared to BR96, for which we could confirm the reported cross-reactivity towards Lewis X,5 and stronger binding of Lewis Y compared to h3S193 as shown by affinity measurement. Further, GT-001 shows no/weak binding to blood cells whereas BR96 and h3S193 significantly bind to different leukocyte subsets. IHC studies reveal that GT-001 stains tumor tissue of different cancer indications (breast cancer, colorectal cancer, head and neck cancer, (non) small cell lung cancer and ovarian cancer) at a high percentage of cases. In ADC surrogate assays, GT-001 potently inhibits the proliferation of several tumor cell lines indicating effective internalization.ConclusionsLewis Y is expressed on many epithelial tumor indications of high medical need. However, several approaches of targeting Lewis Y have failed in the past for efficacy and/or safety reasons. We have developed a humanized antibody that shows superior fine-specificity and higher affinity compared to clinically tested anti-Lewis Y antibodies BR96 and h3S193. Due to the superior fine-specificity, GT-001 shows no/reduced binding of healthy leukocytes potentially reducing side effects as observed for BR96 in the clinic. Its strong target binding and internalization properties make GT-001 an ideal candidate for ADC development.ReferencesAjani JA, Kelsen DP, Haller D, Hargraves K, Healey D. A multi-institutional phase II study of BMS-182248-01 (BR96-doxorubicin conjugate) administered every 21 days in patients with advanced gastric adenocarcinoma. Cancer J 2000;6(2):78–81.Saleh MN, Sugarman S, Murray J, Ostroff JB, Healey D, Jones D, Daniel CR, LeBherz D, Brewer H, Onetto N, LoBuglio AF. Phase I trial of the anti-Lewis Y drug immunoconjugate BR96-doxorubicin in patients with lewis Y-expressing epithelial tumors. J Clin Oncol 2000;18(11):2282–92.Scott AM, Tebbutt N, Lee FT, Cavicchiolo T, Liu Z, Gill S, Poon AM, Hopkins W, Smyth FE, Murone C, MacGregor D, Papenfuss AT, Chappell B, Saunder TH, Brechbiel MW, Davis ID, Murphy R, Chong G, Hoffman EW, Old LJ. A phase I biodistribution and pharmacokinetic trial of humanized monoclonal antibody Hu3s193 in patients with advanced epithelial cancers that express the Lewis-Y antigen. Clin Cancer Res 2007;13(11):3286–92.Smaletz O, Diz MD, do Carmo CC, Sabbaga J, Cunha-Junior GF, Azevedo SJ, Maluf FC, Barrios CH, Costa RL, Fontana AG, Madrigal V, Wainstein AJ, Yeda FP, Alves VA, Moro AM, Blasbalg R, Scott AM, Hoffman EW. A phase II trial with anti-Lewis-Y monoclonal antibody (hu3S193) for the treatment of platinum resistant/refractory ovarian, fallopian tube and primary peritoneal carcinoma. Gynecol Oncol 2015;138(2):272–7.Zhang S, Zhang HS, Cordon-Cardo C, Reuter VE, Singhal AK, Lloyd KO, Livingston PO. Selection of tumor antigens as targets for immune attack using immunohistochemistry: II. Blood group-related antigens. Int J Cancer 1997;73(1):50–6.

A Novel Bispecific Antibody Targeting CD3 and Lewis Y with Potent Therapeutic Efficacy against Gastric Cancer

Lewis Y antigen, a glycan highly expressed on most epithelial cancers, was targeted for cancer treatment but lacked satisfactory results in some intractable and refractory cancers. Thus, it is highly desirable to develop an effective therapy against these cancers, hopefully based on this target. In this work, we constructed a novel T cell-engaging bispecific antibody targeting Lewis Y and CD3 (m3s193 BsAb) with the IgG-[L]-scfv format. In vitro activity of m3s193 BsAb was evaluated by affinity assay to target cells, cytotoxicity assay, cytokines releasing assay, and T cells proliferation and recruiting assays. Anti-tumor activity against gastric cancer was evaluated in vivo by subcutaneous huPBMCs/tumor cells co-grafting model and huPBMCs intravenous injecting model. In vitro, m3s193 BsAb appeared to have a high binding affinity to Lewis Y positive cells and Jurkat cells. The BsAb showed stronger activity than its parent mAb in T cell recruiting, activation, proliferation, cytokine release, and cytotoxicity. In vivo, m3s193 BsAb not only demonstrated higher therapeutic efficacy in the huPBMCs/tumor co-grafting gastric carcinoma model than the parent mAb but also eliminated tumors in the model of intravenous injection with huPBMCs. Strong anti-tumor activity of m3s193 BsAb revealed that Lewis Y could be targeted in T cell-engaging BsAb for gastric cancer therapy.

Gestión del cambio organizacional por covid-19 en las cooperativas de la ciudad de Pilar

La investigación tuvo como objetivo describir el Cambio Organizacional por covid-19 en las Cooperativas de la ciudad de Pilar, en el año 2020. Al respecto, se estudiaron variables relacionadas con debilidades en las estrategias organizacionales y actitudes frente al proceso de cambio organizacional. Se realizó un diagnóstico que permitió proponer un modelo de cambio en la gestión. El estudio adopta una metodología descriptiva-propositiva, trasversal, con enfoque cualitativo, en una población de cinco cooperativas con sus respectivos gerentes, y sesenta funcionarios distribuidos en cada entidad. Como instrumento de recolección de datos se utilizó el cuestionario “Escala de Aptitudes y Resistencia al Cambio” y una entrevista dirigida a los gerentes, que consta de preguntas formuladas con base en las teorías clásicas de Cambio Organizacional de Lewis y Resistencia al Cambio de Chiavenato. Se ha encontrado que las estrategias implementadas son débiles y afectan la manera de trabajar, traduciéndose en una actitud negativa y resistencia frente al proceso de trasformación. Se plantea un modelo de gestión de cambio para mejorar la adaptabilidad de la organización al contexto covid-19.

Expression of the Carbohydrate Lewis Antigen, Sialyl Lewis A, Sialyl Lewis X, Lewis X, and Lewis Y in the Placental Villi of Patients With Unexplained Miscarriages

BackgroundLewis antigens such as Sialyl Lewis A (sLeA), Sialyl Lewis X (sLeX), Lewis X (LeX), and Lewis Y (LeY) are a class of carbohydrate molecules that are known to mediate adhesion between tumor cells and endothelium by interacting with its selectin ligands. However, their potential role in miscarriage remains enigmatic. This study aims to analyze the expression pattern of sLeA, sLeX, LeX, and LeY in the placental villi tissue of patients with a medical history of unexplained miscarriages.MethodsParaffin-embedded slides originating from placental tissue were collected from patients experiencing a miscarriage early in their pregnancy (6–13 weeks). Tissues collected from spontaneous (n = 20) and recurrent (n = 15) miscarriages were analyzed using immunohistochemical and immunofluorescent staining. Specimens obtained from legally terminated normal pregnancies were considered as control group (n = 18). Assessment of villous vessel density was performed in another cohort (n = 10 each group) of gestation ages-paired placenta tissue. Protein expression was evaluated with Immunoreactive Score (IRS). Statistical analysis was performed by using Graphpad Prism 8.ResultsExpression of sLeA, sLeX, LeX, and LeY in the syncytiotrophoblast was significantly upregulated in the control group compared with spontaneous and recurrent miscarriage groups. However, no prominent differences between spontaneous and recurrent miscarriage groups were identified. Potential key modulators ST3GAL6 and NEU1 were found to be significantly downregulated in the recurrent miscarriage group and upregulated in the spontaneous group, respectively. Interestingly, LeX and LeY expression was also detected in the endothelial cells of villous vessels in the control group but no significant expression in miscarriage groups. Furthermore, assessment of villous vessel density using CD31 found significantly diminished vessels in all size groups of villi (small villi <200 µm, P = 0.0371; middle villi between 200 and 400 µm, P = 0.0010 and large villi >400 µm, P = 0.0003). Immunofluorescent double staining also indicated the co-localization of LeX/Y and CD31.ConclusionsThe expression of four mentioned carbohydrate Lewis antigens and their potential modulators, ST3GAL6 and NEU1, in the placenta of patients with miscarriages was significantly different from the normal pregnancy. For the first time, their expression pattern in the placenta was illustrated, which might shed light on a novel understanding of Lewis antigens’ role in the pathogenesis of miscarriages.

Fucosylated CD147 promotes the malignant progression of ovarian cancer and is associated with glycolysis

Background This study investigated the molecular structural relationship between CD147 and Lewis y antigen in ovarian cancer cells, and explored the molecular mechanisms by which Lewis y leads to the malignant progression of ovarian cancer. Methods The expression of CD147 and Lewis y in three epithelial ovarian cell lines (RMG-I, COC1 and HO8910) and their sub-lines (RMG-I-H, COCI/DDP and HO8910/PM) with high metastatic potential and chemotherapy resistance was detected by quantitative real-time PCR, immunocytochemistry, and western blotting. The structural relationship between Lewis y and CD147 was determined by immunoprecipitation. Gene expression enrichment analysis was performed to elucidate the possible role of CD147 in the response to Lewis y in ovarian cancer. Results The expression of CD147, Lewis y, and FUT1 mRNA was significantly lower in ovarian cancer cell lines than in cells with a higher malignancy grade. Lewis y was an important component of CD147, and was predominantly expressed in the highly glycosylated form of CD147. Genes associatd with the CD147-mediated response to Lewis y were mainly involved in cytokine-mediated signaling pathways and hexose metabolic processes. The expression of IL1A (IL-1α), which was highest in ovarian cancer, was significantly higher than in borderline, benign and normal ovarian tissues, and it was positively correlated with Lewis y in ovarian cancer. Conclusion CD147 was modified by fucosylation, and the effect of fucosylated CD147 on promoting the malignant progression of ovarian cancer may be related to glycolysis.