Comprehensive Analysis of Nivolumab, A Therapeutic Anti-Pd-1 Monoclonal Antibody: Impact of Handling and Stress

This study was partially funded by Project P20-01029 (I+D+i-Junta de Andalucia, Spain) and by Project B-FQM-308-UGR20 (Universidad de Granada, Proyectos I+D+i del Programa Operativo FEDER Andalucia 2020) which means that it was also partially supported by European Regional Development Funds.Nivolumab, formulated in the medicine Opdivo (R) (10 mg/mL), is a therapeutic monoclonal antibody (mAb) used in the treatment of different types of cancer. Currently, there is insufficient knowledge about the behaviour of this protein with regards to the risk associated with its routine handling or unintentional mishandling, or when subjected to stress conditions in hospitals. These conditions can be simulated in forced degradation studies, which provide an in-depth understanding of the biophysical and biochemical properties of mAbs. In this study, we carried out a physicochemical and functional characterisation of nivolumab, which was subjected to various stress conditions: heat, freeze/thaw cycles, agitation, light exposure and high hypertonic solution. We used a wide range of analytical techniques: Far-UV CD, IT-FS, DLS, SE/UHPLC(UV)-[Native]MS, and ELISA. The results show that exposure to light was the stress test with the greatest impact on the samples, revelling the formation of non-natural dimers and a different isoform profile. In addition, nivolumab (Opdivo (R)) demonstrated stability up to 60 degrees C (1 h). As regards functionality all the nivolumab (Opdivo (R)) stressed samples were found to be stable except for those subjected to light and agitation, and to a lesser extent, those subjected to FTC 5 and NaCl stresses.I+D+i-Junta de Andalucia, Spain P20-01029European Commission B-FQM-308-UGR2

Using street based metrics to characterize urban typologies

[EN] Urban spatial structures reflect local particularities produced during the development of a city. High spatial resolution imagery and LiDAR data are currently used to derive numerical attributes to describe in detail intra-urban structures and morphologies. Urban block boundaries have been frequently used to define the units for extracting metrics from remotely sensed data. In this paper, we propose to complement these metrics with a set of novel descriptors of the streets surrounding the urban blocks under consideration. These metrics numerically describe geometrical properties in addition to other distinctive aspects, such as presence and properties of vegetation and the relationship between the streets and buildings. For this purpose, we also introduce a methodology for partitioning the street area related to an urban block into polygons from which the street urban metrics are derived. We achieve the assessment of these metrics through application of a one-way ANOVA procedure, the winnowing technique, and a decision tree classifier. Our results suggest that street metrics, and particularly those describing the street geometry, are suitable for enhancing the discrimination of complex urban typologies and help to reduce the confusion between certain typologies. The overall classification accuracy increased from 72.7% to 81.1% after the addition street of descriptors. The results of this study demonstrate the usefulness of these metrics for describing street properties and complementing information derived from urban blocks to improve the description of urban areas. Street metrics are of particular use for the characterization of urban typologies and to study the dynamics of cities.The authors appreciate the financial support provided by the Spanish Ministry of Science and Innovation in the framework of the project CGL2010-19591/BTE, and the data made available by the Spanish Instituto Geográfico Nacional (IGN)Hermosilla, T.; Palomar-Vázquez, J.; Balaguer Beser, ÁA.; Balsa Barreiro, J.; Ruiz Fernández, LÁ. (2014). Using street based metrics to characterize urban typologies. Computers, Environment and Urban Systems. 44:68-79. https://doi.org/10.1016/j.compenvurbsys.2013.12.002S68794

Combined use of UV and MS data for ICH Stability-Indication Method: Quantification and isoforms identification of intact nivolumab

Nivolumab (Opdivo®) is a fully human immunoglobulin G4 isotype approved for the treatment of many cancers. It acts as an immune checkpoint inhibitor by blocking the interaction between PD-1 (Programmed Cell Death Protein 1) – an inhibitory receptor expressed on activated T cells- and its ligands, PD-L1 and PD-L2. The quantification of therapeutic proteins in their medicines and pharmaceutical preparations remains challenging because the protein content, a critical quality attribute, must be rigorously calculated using a validated stabilityindicating method, such as that indicated by the International Conference on Harmonization (ICH) quality guidelines, and this requires the analysis of the drug in the presence of its degraded products. In this work, we present an strategy based on the combined use of the UV and MS data to full file the requirement of the ICH-Q2 (R1) to develop and validated as stability indicated a (RP)UHPLC/UV-(HESI/Orbitrap™)MS method for the quantification of nivolumab in medicinal products. A comparative study of all figures of merit of the method using UV or MS data are shown and discussed. The results show that linearity was similar for the two detectors and was established over a range of 4–45 μg/mL and 1–45 μg/mL for the UV and (HESI/Orbitrap™)MS signals, respectively. The sensitivity of the method was higher when using the (HESI/Orbitrap™)MS signal (0.2 μg/mL) than with the UV(2.0 μg/mL). However, the UV signal provided better accuracy and precision than the (HESI/ Orbitrap™)MS signal, which did not meet the criteria for method robustness and system suitability. In spite of this, the MS signal plays a crucial role in this methodology by obtaining the molecular weight profile of the nivolumab isoforms, so enabling us to propose the glycans profile and detect structural modification due to degradation. The specificity of the method was evaluated by conducting forced degradation tests on samples of nivolumab in medicine form. The aim was to find out whether nivolumab suffers structural modifications when subject to stress. Structural modifications were detected by analysing the MS isoform profile, as changes of this kind promote new isoforms that are not chromatographically separated or detected by the UV signal. In this way, we demonstrated that the (RP)UHPLC/UV-(HESI/Orbitrap™)MS method was capable of detecting nivolumab degradation, and was suitable for use in nivolumab stability studies. Thus, the protein content in the daily surplus of the Opdivo® medicine, stored either at room temperature (20 ◦C) or refrigerated at 4 ◦C, could be tracked for 15 days.FPU18/03131 Ministry of Universities, Spain(P20_01029) Junta de Andalucía (Spain) and European Regional Development Fundspostdoctoral position from the Junta de Andalucía, SpainProject P20-01029 (I + D + i - Junta de Andalucía, Spain) Project B-FQM-308-UGR20 (Universidad de Granada, Proyectos I + D + i del Programa Operativo FEDER Andalucía 2020)CBUA/ Universidad de Granad

Análisis estructurales comprensivos de anticuerpos monoclonales y proteínas de fusión terapéuticos. Aplicaciones en estudios de estabilidad en condiciones reales de uso hospitalario

En la presente Tesis Doctoral, se han desarrollado y puesto a punto estrategias analíticas comprensivas para evaluar distintos atributos críticos de la calidad de proteínas terapéuticas comerciales de naturaleza biotecnológica: anticuerpos monoclonales y proteínas de fusión-Fc, que posteriormente han sido empleadas en estudios de caracterización y estabilidad de las mismas en condiciones de uso hospitalario. Las proteínas estudiadas han sido: dos anticuerpos monoclonales, uno innovador u original, el infliximab, en su medicamento Remicade® (Janssen Biologics B.V) y otro biosimilar de este, el CT-P13, en dos medicamentos bioidénticos entre ellos, Inflectra™ (Pfizer Europe MA EEIG) y Remsima (Celltrion Healthcare Hungary Kft); y una proteína de fusión-Fc, el ziv-aflibercept, en su medicamento innovador Zaltrap® (Bayer AG). Estos medicamentos se encuentran entre los más usados a nivel mundial, y en particular, en los hospitales españoles, por lo que fueron seleccionados para llevar a cabo la investigación presentada en esta Tesis Doctoral.Tesis Univ. Granada.Proyectos de investigación financiados por el Instituto de Salud Carlos III FIS PI010-00201 y FIS PI17-0054

Automated extraction of tree and plot-based parameters in citrus orchards from aerial images

A plot-based approach is proposed to detect fruit trees from high spatial resolution aerial images and extract tree and plot-based parameters, such as fraction of tree cover, number of trees, and planting patterns. Each plot image, defined by the boundaries or polygons obtained from a cadastral database, is analyzed independently. The methodology is based on image processing methods: an unsupervised classification with the k-means algorithm is applied, followed by the automatic identification of the classes representing trees. Once extracted, each tree is individualized using a morphological process applied on the binary image of the trees. A set of parameters is calculated at tree and plot levels that produces a comprehensive description of the spectral and morphological aspects of the trees, as well as their spatial distribution in each plot. The methodology was tested on 0.5 m/pixel spatial resolution aerial images of 300 citrus orchard plots which included the three citrus tree typologies found in the Valencia region (Spain). The accuracy of the fruit tree extraction and the parameters calculated was evaluated by comparison with reference data obtained by manual delineation of the images. The automatically extracted fraction of tree cover was significantly related to the reference tree cover area (R2 = 0.96). In the case of the number of detected trees, the R2 values were always higher than 0.90 for the three typologies. Tree location was estimated with an average positional error of 40 cm. The error obtained in the characterization of the planting pattern was less than 50 cm. The proposed methodology may be applied to large agricultural databases, and the derived information combined with precision agricultural techniques could improve the efficiency of various irrigation and agricultural management tasks such as handling per-plot water requirements and distribution.The authors appreciate the financial support provided by the Spanish Ministry of Science and Innovation within the framework of the Projects CGL2009-14220 and CGL2010-19591/BTE, by the Universitat Politecnica de Valencia within the program PAID-06-11, the support of the Spanish Instituto Geografico Nacional (IGN), and the collaboration of the IMIDA and the IVIA through the Telerieg Project (SOE1/P2/E082).Recio Recio, JA.; Hermosilla, T.; Ruiz Fernández, LÁ.; Palomar-Vázquez, J. (2013). Automated extraction of tree and plot-based parameters in citrus orchards from aerial images. Computers and Electronics in Agriculture. 90:24-34. https://doi.org/10.1016/j.compag.2012.10.005S24349

Nuestra Señora del Carmen y San José : un lugar para todos

Participante en la convocatoria: Premios de buenas prácticas en materia de convivencia, Gobierno de Aragón 2009-10Proyecto promovido por el colegio Nuestra Señora del Carmen y San José para la mejora de la convivencia en todo el centro educativo. Sus objetivos son: fomentar la participación de los alumnos en los centros de tiempo libre del barrio; motivar e informar a los padres para que se integren en centros culturales del barrio; dar a conocer las actividades y la realidad del colegio; trabajar conjuntamente con el servicio de menores de la DGA; seguir regularmente la evolución de los casos presentados en la comisión de absentismo de la zona; participar activamente en las actividades que se proponen desde el ayuntamiento y de otras entidades; dar a conocer a los periódicos las actividades que se desarrollan en el colegio; prevenir el consumo de drogas; y potenciar la actividad física. Para ello se crean cuatro planes de acción: la comisión de convivencia, los responsables de absentismo, la normativa establecida en el reglamento de régimen interno y la comisión de pastoral, que enseñan y potencian una serie de normas y valores para una mejor convivencia.Gobierno de Aragón. Departamento de Educación, Cultura y DeporteAragónDirección General de Política Educativa; Avda. Gómez Laguna, 25, planta 2; 50009 Zaragoza; Tel. +34976715416; Fax +34976715496ES

Improved Tear Film Stability in Patients with Dry Eye After Hyaluronic Acid and Galactoxyloglucan Use

Purpose: Artificial tears only provide transient relief for dry eye. To the best of our knowledge, this is the first study to objectively compare treatment with artificial tears with Keratograph 5M, which allows accurate and objective investigation of dry eye and artificial tear treatment. We aimed to evaluate whether a preservative-free combination of 0.4% hyaluronic acid and 0.2% galactoxyloglucan can improve dry eye using the new topographer, Keratograph 5M. Patients and Methods: This prospective longitudinal, single-arm interventional cohort study was performed at a tertiary referral center and involved 20 patients with dry eye (40 eyes). Preservative-free artificial tears were administered every 3 h. The participants under went clinical and instrumental evaluations at baseline, 15, 30, 60, 90 and 120 min after instillation and 1 week and 1 month after treatment. Baseline values were considered as the controls. All patients were assessed with Keratograph 5M for non-invasive Keratograph first break-up time and non-invasive Keratograph average break-up time, conjunctival hyperemia, and tear meniscus height. Ocular surface staining with fluorescein was evaluated using the slit-lamp and fluorescein break-up time examinations, and the Ocular Surface Disease Index score was recorded for each patient. Results: The signs and symptoms improved after 1 month of preservative-free 0.4% hyaluronic acid and 0.2% galactoxyloglucan treatment. There was a significant increase in the non-invasive Keratograph first break-up time and non-invasive Keratograph average break-up time at 15, 30, 60, and 90 min, and 1 week and 1 month (P < 0.05) and a decrease in hyperemia, corneal staining, and Ocular Surface Disease Index scores after 1 week and 1 month (P < 0.05). No treatment-related adverse event was observed. Conclusion: A combination of 0.4% hyaluronic acid and 0.2% galactoxyloglucan artificial tears seems effective for treating dry eye. Keratograph 5M can objectively detect these changes during the follow-up period

Improved Tear Film Stability in Patients with Dry Eye After Hyaluronic Acid and Galactoxyloglucan Use

© 2020 Molina-Solana et al.[Purpose]: Artificial tears only provide transient relief for dry eye. To the best of our knowledge, this is the first study to objectively compare treatment with artificial tears with Keratograph 5M, which allows accurate and objective investigation of dry eye and artificial tear treatment. We aimed to evaluate whether a preservative-free combination of 0.4% hyaluronic acid and 0.2% galactoxyloglucan can improve dry eye using the new topographer, Keratograph 5M. [Patients and Methods]: This prospective longitudinal, single-arm interventional cohort study was performed at a tertiary referral center and involved 20 patients with dry eye (40 eyes). Preservative-free artificial tears were administered every 3 h. The participants underwent clinical and instrumental evaluations at baseline, 15, 30, 60, 90 and 120 min after instillation and 1 week and 1 month after treatment. Baseline values were considered as the controls. All patients were assessed with Keratograph 5M for non-invasive Keratograph first break-up time and non-invasive Keratograph average break-up time, conjunctival hyperemia, and tear meniscus height. Ocular surface staining with fluorescein was evaluated using the slit-lamp and fluorescein break-up time examinations, and the Ocular Surface Disease Index score was recorded for each patient. [Results]: The signs and symptoms improved after 1 month of preservative-free 0.4% hyaluronic acid and 0.2% galactoxyloglucan treatment. There was a significant increase in the non-invasive Keratograph first break-up time and non-invasive Keratograph average break-up time at 15, 30, 60, and 90 min, and 1 week and 1 month (P < 0.05) and a decrease in hyperemia, corneal staining, and Ocular Surface Disease Index scores after 1 week and 1 month (P < 0.05). No treatment-related adverse event was observed. [Conclusion]: A combination of 0.4% hyaluronic acid and 0.2% galactoxyloglucan artificial tears seems effective for treating dry eye. Keratograph 5M can objectively detect these changes during the follow-up period.IATA and RETICS OFTARED have sponsored the translation of this manuscript. AMG-H, JM-I, ER-d-l-R-F, and MC-M are members of RETICS OFTARED ‘RD16/0008/ 0010ʹ, financed by Instituto de Salud Carlos III, as part of the Plan Nacional I+D+i 2013–2016 and co-financed by the European Union (FEDER/FSE) “Una manera de hacer Europa.