Protein biomarker discovery and fast monitoring for the identification and detection of Anisakids by parallel reaction monitoring (PRM) mass spectrometry

8 páginas, 7 figuras, 1 tablaAnisakids are fish-borne parasites that are responsible for a large number of human infections and allergic reactions around the world. World health organizations and food safety authorities aim to control and prevent this emerging health problem. In the present work, a new method for the fast monitoring of these parasites is described. The strategy is divided in three steps: (i) purification of thermostable proteins from fish-borne parasites (Anisakids), (ii) in-solution HIFU trypsin digestion and (iii) monitoring of several peptide markers by parallel reaction monitoring (PRM) mass spectrometry. This methodology allows the fast detection of Anisakids in < 2 h. An affordable assay utilizing this methodology will facilitate testing for regulatory and safety applications. Significance The work describes for the first time, the Protein Biomarker Discovery and the Fast Monitoring for the identification and detection of Anisakids in fishery products. The strategy is based on the purification of thermostable proteins, the use of accelerated in-solution trypsin digestions under an ultrasonic field provided by High-Intensity Focused Ultrasound (HIFU) and the monitoring of several peptide biomarkers by Parallel Reaction Monitoring (PRM) Mass Spectrometry in a linear ion trap mass spectrometer. The workflow allows the unequivocal detection of Anisakids, in < 2 h. The present strategy constitutes the fastest method for Anisakids detection, whose application in the food quality control area, could provide to the authorities an effective and rapid method to guarantee the safety to the consumersThis work was supported by EU-granted project PARASITE (GA312068), Marie Curie actions (FP7-PEOPLE-2012-IEF, ref. 332274) and by the Ramón Areces Foundation XVII National GrantPeer reviewe

Evaluación del analizador hematológico de impedancia eléctrica BAKER 9120+ con sangre de perro y gato

El objetivo de este trabajo es evaluar el analizador hematológico automático de impedancia eléctrica, Baker 9120+ (Menarini Diagnóstica S.A.), con sangre de perro y gato. La precisión intra e interensayo y la reproducibilidad del Baker 9120+ es excelente para todos los parámetros en ambas especies. El arrastre y la linealidad del analizador hematológico es muy buena para todos los parámetros, a excepción del recuento de plaquetas con sangre felina, debido a la sobreposición de la población eritrocitaria y plaquetar. No es posible determinar las diferentes poblaciones leucocitarias en ninguna de las dos especies, independientemente de la cantidad de hemolizante utilizado. La comparación del Baker 9120+ con otro analizador de impedancia eléctrica en el recuento celular y concentración de hemoglobina, y con el método manual para el valor hematocrito, es buena para todos los parámetros. El recuento de eritrocitos, leucocitos y la concentración de hemoglobina son estables durante las 72 horas postextracción, tanto a temperatura ambiente como en refrigeración. El recuento de plaquetas en sangre de perro desciende rápidamente con diferencias significativas a las 4 horas postextracción en las muestras refrigeradas y a las 24 horas en las muestras mantenidas a temperatura ambiente.An authomated blood cell analyser (Baker 9120+) was evaluated with canine and feline blood. Precision and reproducibility were excelent in both species for all the parameters. Carry-over and linearity was very good for all the parameters, except for the platelet count due to the overlapping between erythrocyte and platelet population. There was not possible to differenciate any of the leukocyte populations with the Baker 9120+. The comparability between the Baker 9120+ and another electric impedance analyzer for blood cell count and hemoglobin concentration and the microhematocrit method for the haematocrit value was good for all the parameters. Red blood cell, white blood cell count and haemoglobin concentration are stable during 72 hours after the extraction at 4 ºC and 25 ºC.Canine platelet count dropped significantly at 4 hours post- extraction in those samples stored at 4 "C and after 24 hours in those samples at room temperature

Horizontal gene transfer and diverse functional constrains within a common replication-partitioning system in Alphaproteobacteria: the repABC operon

<p>Abstract</p> <p>Background</p> <p>The <it>repABC </it>plasmid family, which is extensively present within <it>Alphaproteobacteria</it>, and some secondary chromosomes of the <it>Rhizobiales </it>have the particular feature that all the elements involved in replication and partitioning reside within one transcriptional unit, the <it>repABC </it>operon. Given the functional interactions among the elements of the <it>repABC </it>operon, and the fact that they all reside in the same operon, a common evolutionary history would be expected if the entire operon had been horizontally transferred. Here, we tested whether there is a common evolutionary history within the <it>repABC </it>operon. We further examined different incompatibility groups in terms of their differentiation and degree of adaptation to their host.</p> <p>Results</p> <p>We did not find a single evolutionary history within the <it>repABC </it>operon. Each protein had a particular phylogeny, horizontal gene transfer events of the individual genes within the operon were detected, and different functional constraints were found within and between the Rep proteins. When different <it>repABC </it>operons coexisted in the same genome, they were well differentiated from one another. Finally, we found different levels of adaptation to the host genome within and between <it>repABC </it>operons coexisting in the same species.</p> <p>Conclusion</p> <p>Horizontal gene transfer with conservation of the <it>repABC </it>operon structure provides a highly dynamic operon in which each member of this operon has its own evolutionary dynamics. In addition, it seems that different incompatibility groups present in the same species have different degrees of adaptation to their host genomes, in proportion to the amount of time the incompatibility group has coexisted with the host genome.</p

A transcriptomic approach highlights induction of secondary metabolism in citrus fruit in response to Penicillium digitatum infection

<p>Abstract</p> <p>Background</p> <p>Postharvest losses of citrus fruit due to green mold decay, caused by the fungus <it>Penicillium digitaum</it>, have a considerable economic impact. However, little is known about the molecular processes underlying the response of citrus fruit to <it>P. digitatum</it>.</p> <p>Results</p> <p>Here we describe the construction of a subtracted cDNA library enriched in citrus genes preferentially expressed in response to pathogen infection followed by cDNA macroarray hybridization to investigate gene expression during the early stages of colonization of the fruit's peel by <it>P. digitatum</it>. Sequence annotation of clones from the subtracted cDNA library revealed that induction of secondary and amino acid metabolisms constitutes the major response of citrus fruits to <it>P. digitatum </it>infection. Macroarray hybridization analysis was conducted with RNA from either control, wounded, ethylene treated or <it>P. digitatum </it>infected fruit. Results indicate an extensive overlap in the response triggered by the three treatments, but also demonstrated specific patterns of gene expression in response to each stimulus. Collectively our data indicate a significant presence of isoprenoid, alkaloid and phenylpropanoid biosynthetic genes in the transcriptomic response of citrus fruits to <it>P. digitatum </it>infection. About half of the genes that are up-regulated in response to pathogen infection are also induced by ethylene, but many examples of ethylene-independent gene regulation were also found. Two notable examples of this regulation pattern are the genes showing homology to a caffeine synthase and a berberine bridge enzyme, two proteins involved in alkaloid biosynthesis, which are among the most induced genes upon <it>P. digitatum </it>infection but are not responsive to ethylene.</p> <p>Conclusions</p> <p>This study provided the first global picture of the gene expression changes in citrus fruit in response to <it>P. digitatum </it>infection, emphasizing differences and commonalities with those triggered by wounding or exogenous ethylene treatment. Interpretation of the differentially expressed genes revealed that metabolism is redirected to the synthesis of isoprenes, alkaloids and phenylpropanoids.</p

A Review of Emerging Technologies and Their Acceptance in Higher Education

The pandemic caused by COVID-19 impacted the entire world, but technological progress led to the appearance of new and innovative emerging technologies (ETs). These technologies proved to have a wide potential for use as support in education, but being a new technology, certain complications arose when it came to their application in the educational model. Nowadays, there are many digital technologies, so it is necessary to identify those that can be used in today’s education. Digital technologies have been implemented in all types of sectors, one of them being education, and the use of these technologies translates into significant improvements in educational processes and learning outcomes. Despite this, there is currently little research on the use of ETs as a support in the academic process. This research uses systematic mapping (SM) of the last 5 years, together with the Unified Theory of Acceptance and Use of Technology (UTAUT) model to identify the use and acceptance of ETs in higher education. For the SM, a keyword search string was used in three scientific databases (Scopus, Web of Science, and IEEE Xplore). To apply the UTAUT, a survey was conducted with 120 students on the acceptance of ETs as support in higher education. The results obtained indicate that ETs provide some optimization of educational processes, with greater immersion and application of knowledge when using technologies such as augmented reality, virtual reality, and mobile learning. Likewise, ETs can motivate students, allowing them to reach new academic and professional achievements.This work was financed by the Universidad de Las Américas through project code: TIC.LCC.22.01

Complex regional pain syndrome (CRPS), a review

Complex regional pain syndrome is a chronic and painful condition that affects the quality of life of patients. It is usually triggered by a traumatic event of the soft tissues involving the nervous tissue. Although the factors that cause the syndrome are varied and not well known, different etiopathologic concepts have been proposed to explain the presence of this syndrome, such as autonomic dysfunction and changes in CNS plasticity, among others. The patient characteristically presents pain, sensory abnormalities, vasomotor disturbances in the skin, edema, changes in sweating, and motor alterations. The pain is associated with changes in the autonomic nervous system and has a distal predominance. Since there is no definitive diagnostic test, diagnosis is mainly based on a complete medical history and physical examination. Treatment is multidisciplinary and based on pain relief. Although in most cases evolution is favorable, rapid diagnosis and treatment are recommended to avoid dystrophic stage as much as possibl

Influence of cavity type and size of composite restorations on cuspal flexure

Objectives: The present study examines the influence of cavity type, and size of composite restorations on cuspal flexure due to polymerization shrinkage. Methods: Thirty healthy premolars were selected and divided into two experimental groups. Group 1: Mesial, mesiodistal, and MOD cavities were successively prepared in each tooth by means of the SONICSYS approx system (KaVo ®) using a nº 2 (small) diamond tip. Group 2: The same cavity preparation sequence was followed but a nº 3 (medium) tip was used. Cavity preparations were performed and afterwards restored using the same adhesive system and resin composite. Cuspal displacement was measured 15 min after completion of each type of restoration. Results: Under the experimental conditions used, polymerization shrinkage of composite restorations resulted in an inward deflection of the cusps in all the situations evaluated. The Friedman global test found significant differences according to the cavity type and size (p0.05). Conclusions: The present study demonstrates that significantly higher cuspal deflection is observed in MOD restorations, showing that the degree of dental removal influences the cuspal flexure from polymerization shrinkage of composite restorations

Cod 3M Projections: risk estimation and inputs

The last full assessment for the 3M Cod in the NAFO Scientific Council was carried out in June 2015, giving an advice for two years. Some problems concerning the NAFO 3M cod projections were identified during the 2015 Annual Meeting. Trying to solve these issues, a Workshop was organized by the EU in March 2017. An issue raised by the Fisheries Commission in 2015 is that the risk of exceeding Flim for some of the projection scenarios presented in the assessment was difficult to interpret in light of the overlap in credible intervals of the yields of the various options. To solve this issue, the authors developed a new projection method to perform the 3M cod projections and compute risk, projecting a single catch value instead of a distribution of catches. One of the problems raised by the Scientific Council is how to estimate the inputs to be used in the projections due to the rapid changes in the biology of this stock, especially from 2007. Normally the last three years mean is used as input for these parameters. The changes in the biological parameters cause problems in the projections results as the inputs used for the projected years usually are overestimated, leading to an overestimation of the SSB and the associated TAC for a given F. To solve this issue, the authors suggest using just the last year inputs in the projections. An attempt to add uncertainty in the last year inputs was made, examining the interannual changes observed in past years in the inputs and taking into consideration the correlation between them. The uncertainty and the risk in the results increases considerably and it was suggested to study deeply this method before its implementation. The authors conclude that caution should be exercised concerning the results of the projections for this stock. If uncertainty in the projection inputs is implemented, advice for more than one year will not be accurate due to the issues identified.Executive Agency for Small and Medium-sized Enterprises (EASME)Versión del edito