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This study presents the development of a sandwich ELISA method for gluten detection in foods, using recombinant Fab antibody fragments against gliadin. The Fabs were chemically biotinylated and immobilized on streptavidin-coated plates as capture antibodies, while alkaline phosphatase-conjugated Fabs were used as detection antibodies. Four different gliadin-binding Fabs were tested and the Fab pair Fab8E-4 and Fab-C showed the best compatibility. An indirect sandwich immunoassay, using unmodified Fab8E-4 for capture and Fab-C as the detection antibody, achieved a detection limit of 26 ng/mL of gliadin, corresponding to 10 mg/kg of gluten in foods. No cross-reactivity was observed against 60 gluten-free species commonly used in the food industry. Analysis of 50 commercial products demonstrated consistent results compared to the standard method for gluten detection. The complete lack of cross-reactivity of the developed immunoassay with oat products potentially provides an advantage over other gluten detection systems.

Directed evolution is a pivotal strategy for new antibody discovery, which allowed the generation of high-affinity Fabs against gliadin from two antibody libraries in our previous studies. One of the libraries was exclusively derived from celiac patients’ mRNA (immune library) while the other was obtained through a protein engineering approach (semi-immune library). Recent advances in high-throughput DNA sequencing techniques are revolutionizing research across genomics, epigenomics, and transcriptomics. In the present work, an Oxford Nanopore in-lab sequencing device was used to comprehensively characterize the composition of the constructed libraries, both at the beginning and throughout the phage-mediated selection processes against gliadin. A customized analysis pipeline was used to select high-quality reads, annotate chain distribution, perform sequence analysis, and conduct statistical comparisons between the different selection rounds. Some immunological attributes of the most representative phage variants after the selection process were also determined. Sequencing results revealed the successful transfer of the celiac immune response features to the immune library and the antibodies derived from it, suggesting the crucial role of these features in guiding the selection of high-affinity recombinant Fabs against gliadin. In summary, high-throughput DNA sequencing has improved our understanding of the selection processes aimed at generating molecular binders against gliadin.

The Franciscana (also known as the La Plata River Dolphin) is a small dolphin that lives in the coastal waters of Brazil, Uruguay, and Argentina. This species is considered the most endangered marine mammal in the western South Atlantic Ocean. Anatomic dissection of the digestive system of 19 animals of different ages, including 2 neonates, 12 juveniles, and 5 adults, was performed. Parameters related to length, breadth, weight, and diameter of the digestive viscera were considered in each case. Our results show that the Franciscana dolphin presents differential characteristics in relation to several parts of the digestive system, including, specifically, the tongue, the teeth, the stomach, and the small intestine. Thus, this paper add precious information to the actual knowledge of this vulnerable marine mammal species in order to improve conservation efforts.

Gluten, a group of ethanol-soluble proteins present in the endosperm of cereals, is extensively used in the food industry due to its ability to improve dough properties. However, gluten is also associated with a range of gluten-related diseases (GRDs), such as wheat allergies, celiac disease, and gluten intolerance. The recommended treatment for GRDs patients is a gluten-free diet. To monitor adherence to this diet, it is necessary to develop gluten-detection systems in food products. Among the available methods, immunodetection systems are the most popular due to their simplicity, reproducibility, and accuracy. The aim of this study was to generate novel high-affinity antibodies against gluten to be used as the primary reactant in an enzyme-linked immunosorbent assay (ELISA) test. These antibodies were developed by constructing an immune library from mRNA obtained from two celiac patients with a high humoral response to gluten-related proteins. The resulting library (composed by 1.1x107) was subjected to selection against gliadin using phage display technology. Following several rounds of selection, the Fab-C was selected, and demonstrated good functionality in ELISA tests, presenting a limit of detection of 15 mg/kg for detection of gluten in spiked mixtures and food products. The methodology can discriminate gluten-free products according to the current legislation.

Hen’s egg allergy is the second most common food allergy among infants and young children. The possible presence of undeclared eggs in foods poses a significant risk to sensitized individuals. Therefore, reliable egg allergen detection methods are needed to ensure compliance with food labeling and improve consumer protection. This work describes for the first time the application of phage display technology for the generation of a recombinant antibody aimed at the specific detection of hen’s ovomucoid. First, a single-chain variable fragment (scFv) library was constructed from mRNA isolated from the spleen of a rabbit immunized with ovomucoid. After rounds of biopanning, four binding clones were isolated and characterized. Based on the best ovomucoid-binding candidate SR-G1, an indirect phage enzyme-linked immunosorbent assay (phage-ELISA) was developed, reaching limits of detection and quantitation of 43 and 79 ng/mL of ovomucoid, respectively. The developed ELISA was applied to the analysis of a wide variety of food products, obtaining a good correlation with a commercial egg detection assay used as a reference. Finally, in silico modeling of the antigen-antibody complex revealed that the main interactions most likely occur between the scFv heavy chain and the ovomucoid domain-III, the most immunogenic region of this allergen.

Gluten is the main fraction of wheat proteins. It is widely used in the food industry because of the properties that are generated in the dough, but it is also able to trigger diseases like allergies, autoimmunity processes (such as celiac disease), and intolerances in sensitized persons. The most effective therapy for these diseases is the total avoidance of gluten in the diet because it not only prevents damage but also enhances tissue healing. To ensure the absence of gluten in food products labeled as gluten-free, accurate detection systems, like immunoassays, are required. In this work, four recombinant Fab antibody fragments, selected by phage display technology, were produced and tested for specificity and accuracy against gluten in experimental flour mixtures and commercial food products. A high-affinity probe (Fab-C) was identified and characterized. An indirect ELISA test was developed based on Fab-C that complied with the legal detection limits and could be applied in the assessment of gluten-free diets.

Neuroanatomy is always a challenging topic for veterinary students. It is widely accepted that understanding the anatomy of the central nervous system (CNS) is essential to explain many of the pathological processes that affect the brain. Although its study has varied over time to achieve this goal, in human and veterinary medicine it is difficult to find a teaching method that associates normal anatomy with pathological alterations of the brain. For the first time, we have created an educational tool that combines neuroanatomy and neuropathology, using different magnetic resonance (MR) images as a basis and EspINA software as analyzer, to obtain segmented structures and 3D reconstructions of the dog brain. We demonstrate that this combination is an optimal tool to help anatomists to understand the encephalon, and additionally to help clinicians to recognize illness including a multitude of neurological problems. In addition, we have tried to see whether photogrammetry, which is a common technique in other sciences, for example geology, could be useful to teach veterinary neuroanatomy. Although we still need further investigations, we have been able to generate 3D reconstructions of the whole brain, with very promising results to date.

Marine mammals are divided into three groups, with similar adaptations resulting from their aquatic lifestyle: sirenians, pinnipeds, and cetaceans. The present work focused on the South American fur seal, or Arctocephalus australis, a carnivore included in the pinnipeds group. We assessed whether the anatomical features of the Arctocephalus australis’ respiratory system are comparable to those of other land-carnivores or whether these individuals show anatomical adaptations related to their ability to dive or their breath-holding capacities. We studied 11 cadavers of Arctocephalus australis, which included adult (n = 2) and juvenile (n = 9) individuals, by anatomically dissecting their isolated entire respiratory system. Although it is generally similar to that in land-carnivores, we demonstrated that the Arctocephalus australis’s respiratory apparatus shows several specific characteristics. Therefore, our results are of great importance for clinical diagnostic and wildlife conservation purposes.

The observed increase in the prevalence of gluten-related disorders has prompted the development of novel immunological systems for gluten detection in foodstuff. The innovation on these methods relies on the generation of new antibodies, which might alternatively be obtained by molecular evolution methods such as phage display. This work presents a novel approach for the generation of a Fab library by merging semi-synthetic heavy chains built-up from a pre-existent recombinant antibody fragment (dAb8E) with an immune light chain set derived from celiac donors. From the initial phage population (107 candidates) and after three rounds of selection and amplification, four different clones were isolated for further characterization. The phage Fab8E-4 presented the best features to be applied in an indirect ELISA for the detection of gluten in foods, resulting in improved specificity and sensitivity.

Marine mammals play a critical ecological role as both predator and prey. They are divided into three groups that share similar adaptations to their aquatic life, but that have very different origins and life patterns: sirenians, pinnipeds, cetaceans. The species object of our interest is the South American fur seal or Arctocephalus australis, a carnivore classified within the group of pinnipeds. The objective of the present study was to evaluate whether the anatomical characteristics of the Arctocephalus australis’ digestive system are similar to that of other land-carnivores or if, on the contrary, this species shows anatomical adaptations related to their life in the ocean. The study was carried out on 11 cadavers of the species Arctocephalus australis, made up of two adults and nine juveniles, by means of the anatomical dissection of their entire isolated digestive system. We demonstrate that, with several exceptions, the anatomical characteristics of the digestive system of the Arctocephalus australis are similar to those in other carnivores. Therefore, our data constitute an important contribution for clinical diagnostic and conservation purposes, for both veterinarians and biologists.

Labeling of food allergens in accordance with legal regulations is important to protect the health of allergic consumers. The requirements for detecting allergens in foods involve adequate specificity and sensitivity to identify very small amounts of the target allergens in complex food matrices and processed foods. In this work, one hundred commercial samples were analyzed for walnut detection using three different methods: a sandwich enzyme-linked immunosorbent assay (ELISA) kit based on polyclonal antibodies, a direct ELISA using a recombinant multimeric scFv, and a real time PCR. The most sensitive method was real time PCR followed by sandwich ELISA kit and multimeric scFv ELISA. There was agreement between the three methods for walnut detection in commercial products, except for some heat-treated samples or those that contained pecan. The walnut ELISA kit was less affected by sample processing than was the multimeric scFv ELISA, but there was cross-reactivity with pecan, producing some false positives that must be confirmed by real time PCR. According to the results obtained, 7.0 to 12.6% of samples (depending on the analytical method) contained walnut but did not declare it, confirming there is a risk for allergic consumers. Moreover, there was one sample (3.7%) labelled as containing walnut but that tested negative for this tree nut. Genetic and immunoenzymatic techniques offer complementary approaches to develop a reliable verification for walnut allergen labeling.

Pistachio nuts (Pistacia vera) have been consumed by past and present-day civilizations because of their organoleptic characteristics and potential health benefits. However, they can also produce moderate to severe IgE-mediated reactions in allergic individuals. In this work, we report the isolation of the first recombinant antibodies against pistachio nut, produced without animal immunization, to be used in immunoassays for detection of allergenic pistachio in food products. Several phage display biopanning strategies were evaluated to screen the human-based domain antibody library (dAb) in search for pistachio-specific probes. The clone producing the PVF4 phage-dAb was finally selected, and it does not cross-react with cashew despite the phylogenetic proximity with pistachio. Western blot and matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-TOF/TOF) analysis demonstrated that this clone recognised a unique band of ∼22 kDa related to the basic subunit of pistachio 11S globulin (allergen Pis v 2). The PVF4 phage-dAb allowed detection of pistachio in a food matrix with a limit of detection (LOD) of 3983 mg kg-1 in an indirect phage-enzyme-linked immunosorbent assay (ELISA). The ELISA method developed was used to assess applicability of the PVF4 phage-dAb for analysis of 77 commercial food products.

This paper analyzes the changes in the total factor productivity index of a Spanish hotel chain in the period from 2007 to 2010 with the purpose of identifying efficiency patterns for the chain in a period of financial crisis. The Data envelopment analysis (DEA) Malmquist productivity index was used to estimate productivity change in 38 hotels of the AC chain. Results reveal AC hotels’ efficiency trends and, therefore, their competitiveness in the recession period; they also show the changes experienced in these hotels’ total productivity and its components: technological and efficiency changes. Positive efficiency changes were due to positive technical efficiency rather than technological efficiency. The recession period certainly influenced the performance of AC Hotels, which focused on organizational changes rather than investing in technology.

While cadaver dissection and prosection are deemed quintessential methods for studying anatomy, these methods are attenuated by the flat, 2-D structures incorporated into didactic segments of the anatomy courses that minimize students' ability to comprehend anatomy, leading to constraints in the teaching and learning experience. Several tools, such as 3-D virtual anatomy apps and anatomical models, enhance teaching and students' understanding of gross human anatomy. The Anatomage, a virtual anatomy dissection table (VDT), is a compelling technological development with dissection capabilities and a prosection of the human body and its segments. This study intends to explore incorporating a VDT into a physical therapy anatomy course as an enhanced anatomical learning technique. The methods encompass the inclusion of the VDT as an extra dissection table within the cadaver lab setting. Each group of scholars was required to locate the anatomical configurations and dissections assigned to the topic on the VDT. The anatomy course included four examinations covering the entire human body. The results suggest advancements in knowledge retention following the incorporation of VDT in the cadaver dissection portion of the course. Therefore, we ascertained that the VDT is an exemplary tool to adopt in anatomy courses to aid in the comprehension of complex anatomical structures. Integrating VDT is paramount to the assimilation and application of human anatomy in students' future health careers.

Los estudios de turismo se incorporaron a la Universidad en 1996, con la aprobación de las Directrices Generales de la Diplomatura en Turismo. Al incluirse en las mismas a la Contabilidad como materia troncal, nos hemos planteado mostrar en este trabajo empírico la situación real de esta disciplina en los distintos Planes de Estudio en vigor. Como conclusión general del estudio destacamos la imposibilidad de conseguir el objetivo enunciado relativo a la formación de gestores, al menos en lo que a la Contabilidad se refiere, dado su escaso peso en los Planes de Estudio. Especialmente deplorable resulta la situación de la Contabilidad de Gestión, con una escasísima presencia. Esta conclusión nos ha hecho reflexionar acerca de la importancia, a veces no suficientemente considerada, de los procesos de elaboración y reforma de Planes de Estudio.

Los estudios de turismo se incorporaron a la Universidad en 1996, con la aprobación de las Directrices Generales de la Diplomatura en Turismo. Al incluirse en las mismas a la Contabilidad como materia troncal, nos hemos planteado mostrar en este trabajo empírico la situación real de esta disciplina en los distintos Planes de Estudio en vigor. Como conclusión general del estudio destacamos la imposibilidad de conseguir el objetivo enunciado relativo a la formación de gestores, al menos en lo que a la Contabilidad se refiere, dado su escaso peso en los Planes de Estudio. Especialmente deplorable resulta la situación de la Contabilidad de Gestión, con una escasísima presencia. Esta conclusión nos ha hecho reflexionar acerca de la importancia, a veces no suficientemente considerada, de los procesos de elaboración y reforma de Planes de Estudio.

Los estudios de turismo se incorporaron a la Universidad en 1996, con la aprobación de las Directrices Generales de la Diplomatura en Turismo. Al incluirse en las mismas a la Contabilidad como materia troncal, nos hemos planteado mostrar en este trabajo empírico la situación real de esta disciplina en los distintos Planes de Estudio en vigor. Como conclusión general del estudio destacamos la imposibilidad de conseguir el objetivo enunciado relativo a la formación de gestores, al menos en lo que a la Contabilidad se refiere, dado su escaso peso en los Planes de Estudio. Especialmente deplorable resulta la situación de la Contabilidad de Gestión, con una escasísima presencia. Esta conclusión nos ha hecho reflexionar acerca de la importancia, a veces no suficientemente considerada, de los procesos de elaboración y reforma de Planes de Estudio.

Conventional teaching methods, such as 2-dimensional PowerPoint presentations, are among the most common anatomy teaching tools. However, these methods do not allow depth perception of anatomical structures. Therefore, the purpose of this article is to report the incorporation and student's perception of a 3D anatomy app to an Occupational Therapy class as an enhanced anatomical learning technique. For methods, the Complete 3D Anatomy app was used to teach the upper extremity structures. The students used the Likert scale to answer a survey regarding the different anatomical teaching tools that were integrated into the course. The results indicated that 85% of the students reported the anatomy app was very helpful (85%-39/46), helpful (9%-4/46) or good (2%-1/46) in aiding their comprehension of complex anatomical structures. Similarly, 100% of the students considered the anatomy app to be beneficial. Therefore, we conclude that using the 3D anatomy app was a tool for the comprehensive learning of human anatomy. The selection of the 3D app demonstrates this by graduate students as the second-best teaching tool to learn anatomy. For future courses, we will incorporate and explore the benefits of a personal tablet/iPad with 3D apps for each student, along with a virtual dissection table for the human cadaver laboratory.

Abstract Background The World Health Organization 2022 malaria chemoprevention guidelines recommend providing a full course of antimalarial treatment at pre-defined intervals, regardless of malaria status to prevent illness among children resident in moderate to high perennial malaria transmission settings as perennial malaria chemoprevention (PMC) with sulfadoxine-pyrimethamine (SP). The dhps I431V mutation circulating in West Africa has unknown effect on SP protective efficacy. Methods This protocol is for a three-arm, parallel, double-blinded, placebo-controlled, randomised trial in Cameroon among children randomly assigned to one of three directly-observed treatment groups: (i) Group 1 (n = 450) receives daily artesunate (AS) placebo on days − 7 to -1, then active SP plus placebo amodiaquine (AQ) on day 0, and placebo AQ on days 1 and 2; (ii) Group 2 (n = 250) receives placebo AS on days − 7 to -1, then active SP and AQ on day 0, and active AQ on days 1 and 2; and (iii) Group 3 (n = 200) receives active AS on days − 7 to -1, then placebo SP on day 0 and placebo AQ on days 0 to 2. On days 0, 2, 5, 7, and thereafter weekly until day 28, children provide blood for thick smear slides. Dried blood spots are collected on the same days and weekly from day 28 to day 63 for quantitative polymerase chain reaction (qPCR) and genotype analyses. Discussion Our aim is to quantify the chemopreventive efficacy of SP, and SP plus AQ, and measure the effect of the parasite genotypes associated with SP resistance on parasite clearance and protection from infection when exposed to SP chemoprevention. We will report unblinded results including: (i) time-to-parasite clearance among SP and SP plus AQ recipients who were positive on day 0 by qPCR and followed to day 63; (ii) mean duration of SP and SP plus AQ protection against infection, and (iii) mean duration of symptom-free status among SP and SP plus AQ recipients who were parasite free on day 0 by qPCR. Our study is designed to compare the 28-day follow-up of the new WHO malaria chemoprevention efficacy study protocol with extended follow-up to day 63. Trial registration ClinicalTrials.gov NCT06173206; 15/12/2023.

The purpose of this research was to develop a taxonomy of competencies common across different jobs, occupations, and organizations. To achieve this objective, a definition of competency was constructed and a systematic literature review was conducted, selecting studies that: a) included models with general competencies conceptually close to such definition, b) belonged to the Work and Organizational, and Employability areas, and c) were supported by one or more sources or empirical data. Through this procedure, 23 lists of general competencies were identified, whose content analysis, with a high degree of agreement between two independent reviewers, led to a taxonomy of 36 competencies containing the elements common to all of them. This taxonomy of general competencies is a resource intended to assist in the development of new models and serve as a reference for future competency modeling processes related to the work context.