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A region of approximately one megabase of human Chromosome 12 shows extensive linkage disequilibrium in Utah residents with ancestry from northern and western Europe. This strikingly large linkage disequilibrium block was analyzed with statistical and experimental methods to determine whether natural selection could be implicated in shaping the current genome structure. Extended Haplotype Homozygosity and Relative Extended Haplotype Homozygosity analyses on this region mapped a core region of the strongest conserved haplotype to the exon 1 of the Spinocerebellar ataxia type 2 gene (SCA2). Direct DNA sequencing of this region of the SCA2 gene revealed a significant association between a pre-expanded allele \([(CAG)_8CAA(CAG)_4CAA(CAG)_8]\) of CAG repeats within exon 1 and the selected haplotype of the SCA2 gene. A significantly negative Tajima's D value (−2.20, p < 0.01) on this site consistently suggested selection on the CAG repeat. This region was also investigated in the three other populations, none of which showed signs of selection. These results suggest that a recent positive selection of the pre-expansion SCA2 CAG repeat has occurred in Utah residents with European ancestry., Organismic and Evolutionary Biology

Se presenta una propuesta básica de automatización y utilización de tesauros documentales en entornos distribuidos de recuperación de información mediante servicios web basados en Resource Description Framework (RDF). Por esta razón, se revisan en primer lugar las propuestas de marcado descriptivo aparecidas en los últimos cuatro años para la codificación de tesauros documentales. A continuación, se muestra una arquitectura básica de un servidor de tesauros implementado en Java, y, por último, se repasan los distintos protocolos de comunicación e intercambio entre aplicaciones que se pueden usar para implementar este servicio. El texto se acompaña de la aplicación informática que se ha desarrollado.The article presents a basic proposal for the automation and use of thesauri for information retrieval in distributed environments, through web services based on the Resource Description Framework (RDF) architecture. It begins by reviewing the proposals for descriptive tagging for thesauri coding that have appeared over the past four years. This is followed by a description of the basic architecture of a thesaurus implemented in Java. The article concludes by reviewing different communication and data exchange protocols, together with applications that can be used to implement this service. The text is accompanied by the computer application that has been developed.

Es presenta una proposta bàsica d’automatització i utilització de tesaurus documentals en entorns distribuïts de recuperació d’informació mitjançant serveis web basats en l’arquitectura RDF (resource description framework o marc de descripció de recursos). Per aquest motiu, es revisen, en primer lloc, les propostes d’etiquetatge descriptiu aparegudes en els últims quatre anys per a la codificació de tesaurus documentals. A continuació, es mostra una arquitectura bàsica d’un servidor de tesaurus implementat en Java. I, finalment, es repassen els diversos protocols de comunicació i d’intercanvi de dades entre aplicacions que es poden usar per implementar aquest servei. El text s’acompanya de l’aplicació informàtica que s’ha desenvolupat.The article presents a basic proposal for the automation and use of thesauri for information retrieval in distributed environments, through web services based on the Resource Description Framework (RDF) architecture. It begins by reviewing the proposals for descriptive tagging for thesauri coding that have appeared over the past four years. This is followed by a description of the basic architecture of a thesaurus implemented in Java. The article concludes by reviewing different communication and data exchange protocols, together with applications that can be used to implement this service. The text is accompanied by the computer application that has been developed.

Aim: To estimate the prevalence and factors associated with diabetes among older adults and compare the prevalence rate of a three-round national survey of the Mexican Health and Aging Study (MHAS). Methods: A cross-sectional study was conducted with data obtained from MHAS 2015 ( n = 8167), 2018 ( n = 7854), and 2021 ( n = 8060), which comprised a nationally representative sample of older adults in Mexico. The measures included sociodemographic characteristics and health. A binary logistic regression model was used to identify the association between independent variables and self-reported diabetes. Results: The prevalence of diabetes was 26.3%, 27.7%, and 28.1% in 2015, 2018, and 2021, respectively. This prevalence decreased with age and was higher for female, urban older adults, those with multimorbidity, a lower level of education, and without social security coverage for the three years. Age was associated with a lower possibility of presenting diabetes ([OR = 0.79[0.71-0.89]] and [OR = 0.41[0.33-0.52]] in groups aged 75-84 years and ≥85 years, respectively). Females continue to be more likely to present diabetes than males (OR = 1.39 [95% CI 1.25-1.55]). Older adults living in rural areas are 20% less likely to present diabetes than those living in urban areas (OR = 0.80 [95% CI 0.69-0.93]). Uninsured older adults (OR = 1.35 [95% CI 1.20-1.53]), those who wear glasses (OR = 1.23 [95% CI 1.16-1.30]), those with multimorbidity (OR = 1.13 [95% CI 1.01-1.27]), and those who currently drink alcohol (OR = 1.12 [95% CI 1.00-1.25]) were significantly more likely to have diabetes. Conclusion: An elevated prevalence of diabetes was found in older adults in Mexico, while not having access to social security was associated with a higher possibility of presenting diabetes and living in a rural area was associated with a lower possibility of presenting diabetes. Detection, prevention, and control programs should be implemented to reduce the incidence and severity of the disease in older adults and, thus, prevent its associated complications., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2024 Alvaro García Pérez et al.)

The health-promoting antioxidant properties of virgin olive oil (VOO) are today considered priority targets in the new olive breeding programs. Given that these properties depend mainly on its phenolic fraction, whose origin lies in the phenolic compounds present in olive fruit, the objective of this study was to provide further insight into the accumulation dynamics of the main antioxidant compounds, including both polar phenolics and lipophilic tocopherols, during the ontogeny of the olive fruit. Data obtained show that, albeit with significant differences, all the studied genotypes share just after fruit set an intense increase in the synthesis of tyrosol and hydroxytyrosol derivatives, by far the main phenolic compounds of the olive fruit, and a subsequent steady decrease along fruit development and ripening. The accumulation dynamics of flavonoids and tocopherols were different from those of tyrosol and hydroxytyrosol derivatives, presenting a peak of synthesis just before the onset of fruit ripening, and then in general, their content decreases throughout the ripening phase. In the case of flavonoids, all genotypes also share a strong increase in the accumulation of anthocyanins in the final stages of fruit ripening, coinciding with the change in fruit color. Furthermore, the results during the fruit ripening process evidenced that the content of tyrosol and hydroxytyrosol derivatives and tocopherols in the fruit largely determines the content of these groups of compounds in the oil. The information acquired could be useful for the selection of the most suitable moment in the ontogeny of the olive fruit for the search for key genes in the biosynthesis of phenolic compounds.

Objective: To examine the association between low life satisfaction with past-year dental visits in a rural-urban national sample of 50-year-old Mexican adults with diabetes mellitus (DM)., Methods: Data are drawn from the Mexican Health and Aging Study (MHAS), a cross-sectional study conducted in 2018 involving 3,592 older adults aged 50 years and older and living in urban and rural areas in Mexico. Life satisfaction was measured using the Satisfaction with Life Scale (SWLS) and past-year dental visits dichotomized as none and ≥1 dental visits. The Poisson regression analyses were used to assess the association, adjusting for confounders., Results: 62.9% were women, mean age was 65.5 (±9.6), and 16.5% lived in a rural area, while the female subjects continue to present a higher probability of visiting a dentist (PR = 1.28 (95% CI 1.08-1.51)). In terms of age, the ≥70-year group presented 28% lower possibility of visiting a dentist (PR = 0.72 (95% CI 0.60-0.86)). The interaction showed that adults ≥50 years who reside in a rural area and have low life satisfaction were 40% less likely to have visited a dentist in the last year (PR = 0.60 (95% CI 0.37-0.98)) than adults ≥50 years who reside in an urban area and have high life satisfaction., Conclusions: The present study highlights the association between low life satisfaction and past-year dental visits in rural populations. Therefore, rurality should be considered a possible confounder in analysis of life satisfaction in the older adult population., Competing Interests: The authors declare no conflict of interest., (Copyright © 2023 Alvaro García Pérez et al.)

The phenolic compounds containing hydroxytyrosol are the minor components of virgin olive oil (VOO) with the greatest impact on its functional properties and health benefits. Olive breeding for improving the phenolic composition of VOO is strongly dependent on the identification of the key genes determining the biosynthesis of these compounds in the olive fruit and also their transformation during the oil extraction process. In this work, olive polyphenol oxidase (PPO) genes have been identified and fully characterized in order to evaluate their specific role in the metabolism of hydroxytyrosol-derived compounds by combining gene expression analysis and metabolomics data. Four PPO genes have been identified, synthesized, cloned and expressed in Escherichia coli , and the functional identity of the recombinant proteins has been verified using olive phenolic substrates. Among the characterized genes, two stand out: (i) OePPO2 with its diphenolase activity, which is very active in the oxidative degradation of phenols during oil extraction and also seems to be highly involved in the natural defense mechanism in response to biotic stress, and (ii) OePPO3 , which codes for a tyrosinase protein, having diphenolase but also monophenolase activity, which catalyzes the hydroxylation of tyrosol to form hydroxytyrosol.

Fusarium oxysporum f. sp. fragariae and Macrophomina phaseolina are soilborne fungi leading impactful economical losses to strawberry growers worldwide. Symptoms caused by both pathogens are very similar and include vascular discoloration, wilting, stunting, and dieback of plants, but no fruit damage. An extraction of phenolic and volatile compounds was performed on strawberry fruits from three different cultivars while being grown in a plant growth medium infested by each pathogen. Inoculated plants showed higher content of certain phenolic compounds which have antifungal and antioxidant activity and may have a positive impact on strawberry shelf life. On the other hand, root and vascular infections caused by F. oxysporum and M. phaseolina were able to significantly alter strawberry aroma by reducing or increasing the content of specific volatile compounds which also have an important impact on fruit quality. The changes induced in the aroma profiles of the three strawberry cultivars do not only have organoleptic and economic implications for strawberry growers but play an important role in the plant defense system against pathogens. The results indicate a potential of this line of research to develop new tools for the detection and control of soil pathogens., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Diabetes mellitus (DM) is a metabolic disorder whose prevalence has continuously increased worldwide and is associated with dysfunction of the autonomic nervous system and, in particular, that of the sympathetic nervous system (SNS). The objective of this study was to analyze the interaction of DM and the SNS, building a model of sympathectomized diabetic rats to determine alterations in the content of CA (catecholamines) in different intra-abdominal organs. Sympathectomy was conducted with guanethidine (GNT). Additionally, DM was induced with STZ (Streptozotocin). Treatment with GNT decreased norepinephrine (NE) content in all analyzed tissues, with significant differences found in the paraganglia, liver, pancreas, duodenum, and heart compared to the control group. With respect to epinephrine (E), which was only found in the liver, pancreas, and heart, presenting significant differences (p < 0.05) in the heart, a decrease in its concentration was observed for all of the experimental groups with respect to the control. The decrease in dopamine (DA) content due to the GNT−STZ treatment was 30.1% in the heart with respect to the diabetic (STZ) group. The amount of CA in the adrenal medulla indicates the effect of sympathectomy on the GNT group where there was a significant reduction (p < 0.05) of DA. These findings suggest that the elimination of the sympathetic nervous system in diabetic organisms contributed to a decrease in blood glucose; likewise, an alteration in the levels of CA was observed in the different selected organs, possibly attributed to the severity, duration, and pathogenesis of the complications of acute and chronic DM.

Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article.

The quality of virgin olive oil (VOO) is largely determined by the technology used in the industrial process of extracting the oil. Technological innovations within this field aim to strike a proper balance between oil yield and the optimal chemical composition of VOO. The application of pulsed electric fields (PEF) that cause the electroporation of the plant cell membranes favors a more efficient breakage of the olive fruit tissue, which in turn could facilitate the extraction of the oil and some of its key minor components. Pilot-scale and industrial extraction tests have been conducted to assess the effect of PEF technology on the oil extraction yield and on the organoleptic and functional quality of VOO. The best results were obtained by combining the PEF treatment (2 kV/cm) with short malaxation times and a low processing temperature. Under these conditions, PEF technology could decisively improve the oil yield by up to 25% under optimal conditions and enhance the incorporation of phenolic and volatile compounds into the oils. The PEF treatment neither affected the physicochemical parameters used to determine the commercial categories of olive oils, nor the tocopherol content. Similarly, the sensory evaluation of the PEF-extracted oils by means of a panel test did not detect the appearance of any defect or off-flavor. In addition, the intensity of positive attributes (fruity, bitter and pungent) was generally higher in PEF oils than in control oils.

We integrated a magnetohydrodynamic fluid extractor with an amperometric glucose biosensor to develop a wearable device for non-invasive glucose monitoring. Reproducible fluid extraction through the skin and efficient transport of the extracted fluid to the biosensor surface are prerequisites for non-invasive glucose monitoring. We optimized the enzyme immobilization and the interface layer between the sensing device and the skin. The monitoring device was evaluated by extracting fluid through porcine skin followed by glucose detection at the biosensor. The biosensor featured a screen-printed layer of Prussian Blue that was coated with a layer containing glucose oxidase. Both physical entrapment of glucose oxidase in chitosan and tethering of glucose oxidase to electrospun nanofibers were evaluated. Binding of glucose oxidase to nanofibers under mild conditions provided a stable biosensor with analytical performance suitable for accurate detection of micromolar concentrations of glucose. Hydrogels of varying thickness (95-2000 μm) as well as a thin (30 μm) nanofibrous polycaprolactone mat were studied as an interface layer between the biosensor and the skin. The effect of mass transfer phenomena at the biosensor-skin interface on the analytical performance of the biosensor was evaluated. The sensing device detected glucose extracted through porcine skin with an apparent (overall) sensitivity of -0.8 mA/(M·cm 2 ), compared to a sensitivity of -17 mA/(M·cm 2 ) for measurement in solution. The amperometric response of the biosensor correlated with the glucose concentration in the fluid that had been extracted through porcine skin with the magnetohydrodynamic technique., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Bactrocera oleae , the olive fruit fly, is one of the most important pests affecting the olive fruit, causing serious quantitative and qualitative damage to olive oil production. In this study, the changes induced by B. oleae infestation in the biosynthesis of volatile and phenolic compounds in olive (cvs. Picual, Manzanilla, and Hojiblanca) have been analyzed. Despite cultivar differences, the oils obtained from infested fruits showed a significant increase in the content of certain volatile compounds such as ( E) -hex-2-enal, ethanol, ethyl acetate, and β-ocimene and a drastic decrease of the phenolic contents. The impact of those changes on the inferred quality of the oils has been studied. In parallel, the changes induced by the attack of the olive fly on the expression of some key genes in the biosynthesis of volatile and phenolic compounds, such as lipoxygenase, β-glucosidase, and polyphenol oxidase, have been analyzed. The strong induction of a new olive polyphenol oxidase gene ( OePPO2) explains the reduction of phenolic content in the oils obtained from infested fruits and suggest the existence of a PPO-mediated oxidative defense system in olives.

Mechanical treatments can be simple and feasible methods for enhancing the anaerobic digestion of lignocellulosic substrates. This work aims to relate the direct effect of five different mechanical treatments, i.e., variation in the size and number of particles, with the variations in the chemical composition and, subsequently, the effect over the anaerobic digestion of residual raspberry extrudate, which was used as a model substrate. A high variation in the number of particles and the particle size distribution was achieved depending on the mechanical treatment applied, reaching the highest number of particles for the treatments with knife mills and mortar (around 8000 particles per gram). The higher number of particles was related to higher solubilisation, including phenolic compounds and sugars. The combination of knife mills and mortar pretreatment, which presented the highest number of particles, resulted in a 66% more of polyphenols in comparison to the raw substrate. However, the presence of anthocyanins was higher in mechanical treatments with less effect. The enhancement of the anaerobic digestion was clearly related to the increment in the number of particles of small size after the mechanical treatments. The highest methane yield coefficient (236 ± 11 mL CH 4 /g volatile solids) was achieved for the raspberry extrudate treated with knife mills., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: To analyse the effect of secukinumab on self-reported variables of patients diagnosed with psoriatic arthritis and/or ankylosing spondylitis in relation to their health status, pain, fatigue, sleep and quality of life., Methods: A six-month, observational, longitudinal, prospective, multicentre study was conducted with 39 patients who initiated treatment with secukinumab as therapy for psoriatic arthritis and/or spondylitis. The main variables were changes in patient-reported measures and they were evaluated by means of the questionnaires: FACIT-fatigue, Insomnia Severity Index, EuroQol-3L-5D and PsAQoL. In addition, depending on the type of disease (peripheral psoriasis or spondyloarthritis) the DAS28 with ESR or the BASDAI were calculated, respectively., Results: Levels of fatigue, moderate and severe insomnia significantly reduced after 6 months of treatment with secukinumab. At the same time, patient-reported quality of life increased significantly (P = .006). Data on pain and discomfort also show significant improvement after the treatment., Conclusions: Patients with psoriatic arthritis and/or ankylosing spondylitis who start treatment with secukinumab show improvement at 6 months in all effect sizes of the treatment, particularly in sleep, fatigue and quality of life. Furthermore, patient-reported outcome measures are of additional clinical value and allow more accurate and closer assessment of their real status of health and well-being., (Copyright © 2020 Elsevier España, S.L.U. and Sociedad Española de Reumatología y Colegio Mexicano de Reumatología. All rights reserved.)

Virgin olive oil (VOO) is one of the most emblematic products of the Mediterranean diet. Its content in phenolic compounds is strongly associated with the antioxidant and health-promoting properties of this diet. VOO's phenolic profile is determined mainly by the phenolic compounds present in the olive fruit, so knowing their content allows for a fairly precise estimate of the antioxidant and functional properties of the corresponding oil. In this sense, a convenient, green, and sensitive spectrophotometric method was developed for the quantitative determination of total phenolic compounds in olive fruits. The method is based on an easy-to-use extraction procedure of olive fruit phenolics using dimethyl sulfoxide and quantification with the Folin-Ciocalteu reagent. Oleuropein proved to be a suitable reference compound for quantification, displaying a good linear response (r = 0.9996) over the concentration range of 0.58-6.48 mg/mL, with a variation coefficient of 0.42% and limits of detection and quantification of 0.0492 and 0.1490 mg/mL, respectively. The method was validated using a wide array of fruit samples representative of the Olea europaea L. genetic diversity. The results obtained with this spectrophotometric method, expressed as mg/mL of oleuropein, showed a good correlation with those obtained with the fruit samples analyzed by high performance liquid chromatography, with an r value of 0.9930 and a slope value of 1.022, confirming its reliability. Thus, this method can become a very useful simple tool to estimate the total phenolic content of olive fruits, especially when working with numerous samples such as in olive breeding programs or in commercial olive production, in which it is especially useful to know the phenolic state of the fruit and thus determine the optimal harvest date or the most appropriate agronomic treatment to increase the functional properties of the olive fruit and the olive oil.

Background: Hydrolysis of the fruit phenolic glucosides occurring during the oil extraction process is the main biochemical reaction affecting the biosynthesis and accumulation of secoiridoid compounds in virgin olive oil. An integrated approach at the molecular, biochemical, and metabolic level was used to study the olive β-glucosidase gene family in seven olive cultivars selected by their different phenolic profiles., Results: Eight β-glucosidase genes have been identified by in silico analysis of an olive transcriptome. Their expression levels were analyzed by reverse transcription quantitative polymerase chain reaction in olive fruits at different ripening stages: I, green fruits, 16-19 weeks after flowering (WAF); II, yellow-green fruits, 22-25 WAF; III, turning fruits, 28-31 WAF; and IV, fully ripe fruits, 35-40 WAF. Gene expression was compared with the level of β-glucosidase activity in the fruit and with the phenolic composition of fruits and oils from different olive cultivars. Phylogenetic analysis of the encoded proteins and differences found among the β-glucosidase genes based on Gene Ontology enrichment analysis data suggests maximum involvement of two genes, OeBGLU1A and OeBGLU1B, in the phenolic composition of virgin olive oil. Positive correlation coefficients were found within each olive cultivar between OeBGLU1A and OeBGLU1B gene expression data and the phenolic content of the oil., Conclusion: The results obtained suggest that the expression pattern of specific β-glucosidase genes may be an accurate predictor for the phenolic content of virgin olive oil that could be used in olive breeding programs. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Hydroxytyrosol derivatives are the most important phenolic components in virgin olive oil due to their well-demonstrated biological activities. In this regard, two phenyl acetaldehyde reductase genes, OePAR1.1 and OePAR1.2, involved in hydroxytyrosol synthesis, have been identified from an olive transcriptome. Both genes were synthesized and expressed in Escherichia coli , and their encoded proteins were purified. The recombinant enzymes display high substrate specificity for 2,4-dihydroxyphenylacetaldehyde (3,4-DHPAA) to form hydroxytyrosol. The reaction catalyzed by OePAR constitutes the second, and last, biochemical step in the formation of hydroxytyrosol from the amino acid L-3,4-dihydroxyphenylalanine (L-DOPA) in olive. OePAR1.1 and OePAR1.2 enzymes exhibit high thermal stability, similar pH optima (pH 6.5), and high affinity for 3,4-DHPAA (apparent K m 0.6 and 0.8 µmol min -1 mg -1 , respectively). However, OePAR1.2 exhibited higher specific activity and higher expression levels in all the olive cultivars under study. The expression analyses indicate that both OePAR1.1 and OePAR1.2 genes are temporally regulated in a cultivar-dependent manner. The information provided here could be of interest for olive breeding programs searching for new olive genotypes with the capacity to produce oils with higher levels of hydroxytyrosol derivatives.

Changes in protein levels in different components of the apical junctional complex occur in colorectal cancer (CRC). Claudin‑3 is one of the main constituents of tight junctions, and its overexpression can increase the paracellular flux of macromolecules, as well as the malignant potential of CRC cells. The aim of this study was to investigate the molecular mechanisms involved in the regulation of claudin‑3 and its prognostic value in CRC. In silico evaluation in each of the CRC consensus molecular subtypes (CMSs) revealed that high expression levels of CLDN3 (gene encoding claudin‑3) in CMS2 and CMS3 worsened the patients' long‑term survival, whereas a decrease in claudin‑3 levels concomitant with a reduction in phosphorylation levels of epidermal growth factor receptor (EGFR) and insulin‑like growth factor 1 receptor (IGF1R) could be achieved by inhibiting N‑glycan biosynthesis in CRC cells. We also observed that specific inactivation of these receptor tyrosine kinases (RTKs) led to a decrease in claudin‑3 levels, and this regulation seems to be mediated by phospholipase C (PLC) and signal transducer and activator of transcription 3 (STAT3) in CRC cells. RTKs are modulated by their N‑linked glycans, and inhibition of N‑glycan biosynthesis decreased the claudin‑3 levels; therefore, we evaluated the correlation between N‑glycogenes and CLDN3 expression levels in each of the CRC molecular subtypes. The CMS1 (MSI immune) subtype concomitantly exhibited low expression levels of CLDN3 and N‑glycogenes (MGAT5, ST6GAL1, and B3GNT8), whereas CMS2 (canonical) exhibited high gene expression levels of CLDN3 and N‑glycogenes (ST6GAL1 and B3GNT8). A robust positive correlation was also observed between CLDN3 and B3GNT8 expression levels in all CMSs. These results support the hypothesis of a mechanism integrating RTK signaling and N‑glycosylation for the regulation of claudin‑3 levels in CRC, and they suggest that CLDN3 expression can be used to predict the prognosis of patients identified as CMS2 or CMS3.

The phenolic composition of virgin olive oil (VOO) is strongly determined by the content and distribution of secoiridoid phenolic glucosides present in the olive fruit. Among them, oleuropein is the most abundant in olive mesocarp and is characterized by containing an hydroxytyrosol residue in its chemical structure. Hydroxytyrosol-containing molecules are those that exhibit the most important biological activities in virgin olive oil. In this regard, we identified an aromatic aldehyde synthase gene ( OeAAS ) from an olive transcriptome, which was synthesized, expressed in Eschrichia coli , and purified its encoded protein. The recombinant OeAAS is a bifunctional enzyme catalyzing decarboxylation and amine-oxidation reactions in a single step. OeAAS displays strict substrate specificity for l-DOPA to form 2,4-dihydroxyphenylacetaldehyde, the immediate precursor of hydroxytyrosol. In addition to the biochemical characterization of the enzyme, the expression analysis carried out in different olive cultivars and ripening stages indicate that OeAAS gene is temporally regulated in a cultivar-dependent manner. High correlation coefficients were found between OeAAS expression levels and the phenolic content of olive fruits and oils, which supports a key role for OeAAS in the accumulation of hydroxytyrosol-derived secoiridoid compounds in olive fruit and virgin olive oil.

Virgin olive oil (VOO) is the main source of lipids in the Mediterranean diet and one of the main contributors to its proven protection against diseases associated with chronic inflammation states. This oil is rich in antioxidant compounds such as tocopherols, which together constitute the vitamin E stock of the oil. The purpose of the present work was to conduct a study on the diversity of the contents of vitamin E in the olive species ( Olea europaea L.), and to know how the season climatic conditions and the degree of fruit ripening stage influences the final content of this vitamin in VOO. Data showed that the content of vitamin E in VOO is highly dependent on the olive cultivar, displaying a wide variability (89-1410 mg tocopherol/kg oil) in the olive species, and that is also dependent, to a lesser extent, on the crop year climate and the stage of fruit ripening. In addition, the suitability of cultivar crosses for breeding programs to obtain new cultivars with improved vitamin E content in VOO has been assessed. Our findings demonstrated that a single cross of olive cultivars may provide sufficient variability to be used in the selection of new cultivars.

To understand in-depth material properties, manufacturing, and conservation in cultural heritage artefacts, there is a strong need for advanced characterization tools that enable analysis down to the nanometric scale. Transmission electron microscopy (TEM) and electron diffraction (ED) techniques, like 3D precession electron diffraction tomography and ASTAR phase/orientation mapping, are proposed to study cultural heritage materials at nanoscale. In this work, we show how electron crystallography in TEM helps to determine precise structural information and phase/orientation distribution of various pigments in cultural heritage materials from various historical periods like Greek amphorisks, Roman glass tesserae, and pre-Hispanic Maya mural paintings. Such TEM-based methods can be an alternative to synchrotron techniques and can allow distinguishing accurately different crystalline phases even in cases of identical or very close chemical compositions at the nanometric scale.

Verticillium wilt, caused by the defoliating pathotype of Verticillium dahliae , is the most devastating soil-borne fungal disease of olive trees, and leads to low yields and high rates of tree mortality in highly susceptible cultivars. The disease is widely distributed throughout the Mediterranean olive-growing region and is one of the major limiting factors of olive oil production. Other than effects on crop yield, little is known about the effect of the disease on the content of volatile compounds and phenolics that are produced during the oil extraction process and determine virgin olive oil (VOO) quality and commercial value. Here, we aim to study the effect of Verticillium wilt of the olive tree on the content of phenolic and volatile compounds related to the sensory properties of VOO. Results showed that synthesis of six and five straight-chain carbon volatile compounds were higher and lower, respectively, in oils extracted from infected trees. Pathogen infection affected volatile compounds known to be contributors to VOO aroma: average content of one of the main positive contributors to VOO aroma, ( E )-hex-2-enal, was 38% higher in oils extracted from infected trees, whereas average content of the main unpleasant volatile compound, pent-1-en-3-one, was almost 50% lower. In contrast, there was a clear effect of pathogen infection on the content of compounds responsible for VOO taste, where average content of the main bitterness contributor, oleuropein aglycone, was 18% lower in oil extracted from infected plants, and content of oleocanthal, the main contributor to pungency, was 26% lower. We believe this is the first evidence of the effect of Verticillium wilt infection of olive trees on volatile compounds and phenolics that are responsible of the aroma, taste, and commercial value of VOO.

Objective : To evaluate the impact of Molar Incisor Hypomineralization (MIH) in the OHRQoL in a group of 8 to 10 year-old Mexican schoolchildren. Study design: A cross-sectional study of 411 8 to 10 year-old schoolchildren selected from public schools in Naucalpan, in the State of Mexico. The presence and severity of MIH was evaluated using the European Academy of Pediatric Dentistry (EAPD) criteria. The OHRQoL was evaluated using the Child Perceptions Questionnaire (CPQ). Poisson regression models were performed for the analysis. Results: The prevalence of MIH in the permanent dentition was 40.4%. High scores were found in the four domains of the CPQ for children with MIH compared to children without MIH (p<0.001). In the Poisson model, a greater negative impact in the four CPQ domains was found for schoolchildren with MIH: oral symptoms [RR=1.75 (CI95% 1.63-1.78)]; functional limitations [RR=2.17 (CI95% 2.01-2.35)]; emotional wellbeing [RR=1.94 (CI95% 1.80-2.08)]; and, social well-being [RR=2.62 (CI95% 2.49-2.77)]. A greater impact on the four CPQ domains were found for children with moderate/severe MIH compared to children without MIH (p<0.001). Conclusion: Children with moderate/severe MIH experienced a greater negative impact on their OHRQoL compared to children without MIH.

Olive growing is mainly based on traditional varieties selected by the growers across the centuries. The few attempts so far reported to obtain new varieties by systematic breeding have been mainly focused on improving the olive adaptation to different growing systems, the productivity and the oil content. However, the improvement of oil quality has rarely been considered as selection criterion and only in the latter stages of the breeding programs. Due to their health promoting and organoleptic properties, phenolic compounds are one of the most important quality markers for Virgin olive oil (VOO) although they are not commonly used as quality traits in olive breeding programs. This is mainly due to the difficulties for evaluating oil phenolic composition in large number of samples and the limited knowledge on the genetic and environmental factors that may influence phenolic composition. In the present work, we propose a high throughput methodology to include the phenolic composition as a selection criterion in olive breeding programs. For that purpose, the phenolic profile has been determined in fruits and oils of several breeding selections and two varieties ("Picual" and "Arbequina") used as control. The effect of three different environments, typical for olive growing in Andalusia, Southern Spain, was also evaluated. A high genetic effect was observed on both fruit and oil phenolic profile. In particular, the breeding selection UCI2-68 showed an optimum phenolic profile, which sums up to a good agronomic performance previously reported. A high correlation was found between fruit and oil total phenolic content as well as some individual phenols from the two different matrices. The environmental effect on phenolic compounds was also significant in both fruit and oil, although the low genotype × environment interaction allowed similar ranking of genotypes on the different environments. In summary, the high genotypic variance and the simplified procedure of the proposed methodology for fruit phenol evaluation seems to be convenient for breeding programs aiming at obtaining new cultivars with improved phenolic profile.

The suitability of the cold storage (2°C) of fruit to maintain the quality of 'Manzanilla de Sevilla' and 'Manzanilla Cacereña' intended for virgin olive oil extraction was investigated. This temperature was effective in keeping the best commercial category of oil quality in both manually harvested olives and in mechanically harvested 'Manzanilla Cacereña' fruits for 11days. Mechanical harvesting induced significant decreases in oxidative stability and in the main phenolic compounds contents in the oils during cold storage and, only initially, in the total volatiles, regardless of the cultivar considered. However, the contents of volatile esters, associated to fruity flavor, were always higher in the oils from mechanically harvested fruits. 'Manzanilla de Sevilla' oils exhibited higher total volatiles during fruit cold storage, regardless of the harvesting system used., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Phenolic composition of virgin olive oil is determined by the enzymatic and/or chemical reactions that take place during olive fruit processing. Of these enzymes, β-glucosidase activity plays a relevant role in the transformation of the phenolic glycosides present in the olive fruit, generating different secoiridoid derivatives. The main goal of the present study was to characterize olive fruit β-glucosidase genes and enzymes responsible for the phenolic composition of virgin olive oil. To achieve that, we have isolated an olive β-glucosidase gene from cultivar Picual ( OepGLU ), expressed in Nicotiana benthamiana leaves and purified its corresponding recombinant enzyme. Western blot analysis showed that recombinant OepGLU protein is detected by an antibody raised against the purified native olive mesocarp β-glucosidase enzyme, and exhibits a deduced molecular mass of 65.0 kDa. The recombinant OepGLU enzyme showed activity on the major olive phenolic glycosides, with the highest levels with respect to oleuropein, followed by ligstroside and demethyloleuropein. In addition, expression analysis showed that olive GLU transcript level in olive fruit is spatially and temporally regulated in a cultivar-dependent manner. Furthermore, temperature, light and water regime regulate olive GLU gene expression in olive fruit mesocarp. All these data are consistent with the involvement of OepGLU enzyme in the formation of the major phenolic compounds present in virgin olive oil.

The human epidermal growth factor receptor (HER1) and its partner HER2 are extensively described oncogenes and validated targets for cancer therapy. However, the effectiveness of monospecific therapies targeting these receptors is hampered by resistance emergence, which is frequently associated with the upregulation of other members of HER family. Combined therapies using monoclonal antibodies or tyrosine kinase inhibitors have been suggested as a promising strategy to circumvent this resistance mechanism. We propose an alternative approach based on simultaneous inactivation of HER1 and HER2 by multi-epitope blockade with specific polyclonal antibodies induced by vaccination. Elicited antibodies impaired both receptors activation and induced their degradation, which caused the inhibition of down-signaling cascades. This effect was translated into cell cycle arrest and apoptosis induction of human tumor cells. Elicited antibodies were able to reduce the viability of a panel of human tumor lines with differential expression levels of HER1 and HER2. The most significant effects were obtained in the tumor lines with lower expression levels of both receptors. These new insights would contribute to the rational design of HER receptors targeting multivalent vaccines, as an encouraging approach for the treatment of cancer patients., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest.

Virgin olive oil (VOO) is the only food product requiring official sensory analysis to be classified in commercial categories, in which the evaluation of the aroma plays a very important role. The selection of parents, with the aim of obtaining new cultivars with improved oil aroma, is of paramount importance in olive breeding programs. We have assessed the volatile fraction by headspace-solid-phase microextraction/gas chromatography-mass spectrometry-flame ionization detection (HS-SPME/GC-MS-FID) and the deduced aroma properties of VOO from a core set of olive cultivars (Core-36) which possesses most of the genetic diversity found in the World Olive Germplasm Collection (IFAPA Alameda del Obispo) located in Cordoba, Spain. The VOO volatile fractions of Core-36 cultivars display a high level of variability. It is mostly made of compounds produced from polyunsaturated fatty acids through the lipoxygenase pathway, which confirms to be a general characteristic of the olive species ( Olea europaea L.). The main group of volatile compounds in the oils was six straight-chain carbon compounds derived from linolenic acid, some of them being the main contributors to the aroma of the olive oils according to their odor activity values (OAV). The high level of variability found for the volatile fraction of the oils from Core-36 and, therefore, for the aroma odor notes, suggest that this core set may be a very useful tool for the choice of optimal parents in olive breeding programs in order to raise new cultivars with improved VOO aroma.

Background: To develop a population pharmacokinetic model of gentamicin in children with complicated severe malnutrition and to study the influence of covariates (weight and age) on pharmacokinetic indices. In addition, we use the model to perform Monte Carlo simulations to explore the efficacy of several dosage regimens., Methods: Twenty-six children with severe complicated malnutrition were studied. Ninety-six samples of gentamicin plasma concentrations, obtained from 0.5 to 8 hours after intravenous dosing, were analyzed. Population pharmacokinetic models were built using the program Monolix 4.2 (Lixoft, Antony, France). Monte Carlo simulations were performed to evaluate optimal dosage regimens, using the final pharmacokinetic model, based on the probability of pharmacokinetic-pharmacodynamic target attainment., Results: The concentration-time data were fitted best to 1-compartment model. The estimated population clearance was 1.1 L/h, and the volume of distribution was 2.23 L, with an interindividual variability of 47.2% and 35.6%, respectively. The final models for the clearance and volume of distribution were as follows: CL (L/h) = CL = 1.15 (age/median age) and V (L) = 2.33 (weight/median weight). In Monte Carlo simulations, gentamicin given in dosages of 7.5 to 15 mg/kg optical density was effective in achieving the pharmacodynamic target Cmax:minimal inhibitory concentration >10 for minimal inhibitory concentrations ≤2.5 mg/L, with a probability lower than 1% for Cmin >1 mg/L., Conclusions: Based on the available evidence, an intravenous dose of 7.5 to 15 mg/kg once daily in children with complicated severe malnutrition and normal renal function ensures high probability of efficacy and low risk of nephrotoxicity, which gives further support to the recommendations issued by the World Health Organization treatment for this patient population.

Volatile compounds are responsible for most of the sensory qualities of virgin olive oil and they are synthesized when enzymes and substrates come together as olive fruit is crushed during the industrial process to obtain the oil. Here we have studied the variability among the major volatile compounds in virgin olive oil prepared from the progeny of a cross of Picual and Arbequina olive cultivars (Olea europaea L.). The volatile compounds were isolated by SPME, and analyzed by HRGC-MS and HRGC-FID. Most of the volatile compounds found in the progeny's oil are produced by the enzymes in the so-called lipoxygenase pathway, and they may be clustered into different groups according to their chain length and polyunsaturated fatty acid origin (linoleic and linolenic acids). In addition, a group of compounds derived from amino acid metabolism and two terpenes also contributed significantly to the volatile fraction, some of which had significant odor values in most of the genotypes evaluated. The volatile compound content of the progeny was very varied, widely transgressing the progenitor levels, suggesting that in breeding programs it might be more effective to consider a larger number of individuals within the same cross than using different crosses with fewer individuals. Multivariate analysis allowed genotypes with particularly interesting volatile compositions to be identified and their flavor quality deduced., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The ability of olive endogenous enzymes β-glucosidase, polyphenol oxidase (PPO) and peroxidase (POX), to determine the phenolic profile of virgin olive oil was investigated. Olives used for oil production were stored for one month at 20 °C and 4 °C and their phenolic content and enzymatic activities were compared to those of ripening olive fruits. Phenolic and volatile profiles of the corresponding oils were also analysed. Oils obtained from fruits stored at 4 °C show similar characteristics to that of freshly harvested fruits. However, the oils obtained from fruits stored at 20 °C presented the lowest phenolic content. Concerning the enzymatic activities, results show that the β-glucosidase enzyme is the key enzyme responsible for the determination of virgin olive oil phenolic profile as the decrease in this enzyme activity after 3 weeks of storage at 20 °C was parallel to a dramatic decrease in the phenolic content of the oils., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The aim of this work was to study the content of tocopherols, sterols, triterpenic and aliphatic alcohols, carotenoids, and volatile compounds in the kernel oils from six walnut (Juglans regia L.) varieties. The levels of β-carotene ranged between 0.22 and 0.62 mg/kg, followed by lutein (0.01-0.06 mg/kg). The total content of tocopherol ranged from 186.5 to 436.2 mg/kg of the extracted oil and the major isoform in all samples was γ-tocopherol. The most abundant phytosterol was β-sitosterol (974-1494 mg/kg) followed by campesterol then Δ-5-avenasterol. The major triterpenic alcohol was cycloartenol (226.4-532.1 mg/kg). Hexacosanol (9.71-28.15 mg/kg) was the major aliphatic alcohol. The detected volatile compounds were pentanal, hexanal, nonanal, 2-decenal and hexanol. The statistical analysis showed significant differences between varieties, which are probably due to genetic factors., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The effect of modifying polyphenol oxidase (PPO) and peroxidase (POX) activity during the extraction of virgin olive oil has been assessed in terms of its influence on the phenolic profile of the oil produced. These enzymes were modified by adding exogenous enzyme or specific inhibitors during the milling and subsequent kneading step, studying the effect on specific phenolic compounds in the oils. PPO is the main enzyme involved in phenolic oxidation at the milling step whereas POX activity seems to be the main influence during the kneading step. The data obtained suggest it is possible to increase the nutritional and organoleptic quality of virgin olive oil by inhibiting these enzymes during olive fruit processing. Treatment with the PPO inhibitor tropolone produced a twofold increase in the phenolic fraction, which would therefore seem to be an interesting strategy to improve the nutritional and organoleptic properties of virgin olive oil., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Unlabelled: Oxygen is a neonatal health hazard that should be avoided in clinical practice. In this review, an international team of neonatologists and nurses assessed oxygen saturation (SpO2 ) targeting in preterm infants and evaluated the potential weaknesses of randomised clinical trials., Conclusion: SpO2 of 85-89% can increase mortality and 91-95% can cause hyperoxia and ill effects. Neither of these ranges can be recommended, and wider intermediate targets, such as 87-94% or 88-94%, may be safer., (© 2014 The Authors. Acta Paediatrica published by John Wiley & Sons Ltd on behalf of Foundation Acta Paediatrica.)

2-Phenylethanol (PEA), an important alcohol derived from phenylalanine, is involved in aroma and flavour of bakers' products. Four spontaneous mutants of an industrial bakers' yeast, V1 strain, were isolated for their resistance to p-fluoro-DL-phenylalanine (PFP), a toxic analogue of L-phenylalanine. Mutants overproduced this amino acid and showed variations in their internal pool for several other amino acids. Moreover, a rise in PEA production after growth in industrial medium (MAB) was observed in three of the mutants, although their growth and fermentative capacities were slightly impaired. However, concentration of PEA remained higher during dough fermentation and also after baking, thus improving taste and aroma in bread., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Virgin olive oil phenolic compounds are responsible for its nutritional and sensory quality. The synthesis of phenolic compounds occurs when enzymes and substrates meet as olive fruit is crushed during the industrial process to obtain the oil. The genetic variability of the major phenolic compounds of virgin olive oil was studied in a progeny of the cross of Picual x Arbequina olive cultivars (Olea europaea L.). They belong to four different groups: compounds that included tyrosol or hydroxytyrosol in their molecules, lignans, flavonoids, and phenolic acids. Data of phenolics in the oils showed that the progeny displayed a large degree of variability, widely transgressing the genitor levels. This high variability can be of interest on breeding programs. Thus, multivariate analysis allowed to identify genotypes within the progeny particularly interesting in terms of phenolic composition and deduced organoleptic and nutritional quality. The present study has demonstrated that it is possible to obtain enough degree of variability with a single cross of olive cultivars for compounds related to the nutritional and organoleptic properties of virgin olive oil.

Objective: Mexican Cervical Cancer Screening Study II (MECCS II) seeks to develop a highly sensitive and highly specific screening program able to be adapted to all socioeconomic levels in Mexico. The objectives of MECCS II are (1) to compare the sensitivity and specificity for cervical intraepithelial neoplasia (CIN) 3 or cancer of self-collected vaginal specimens tested for high-risk types of the human papillomavirus (HR-HPV) by APTIMA with those tested for HR-HPV by Hybrid Capture 2 (HC2); and (2) determine the efficacy of cryotherapy in the treatment of HR-HPV-positive and acetic acid-aided visual inspection (VIA)-positive and -negative women after VIA triage., Methods: The study was conducted in rural Mexico. Women aged 30 to 50 years, nonpregnant, with no history of hysterectomy or pelvic irradiation and varied histories of screening, participated. A direct endocervical sample was tested for cytology, HC2, and APTIMA assay (AHPV). Subjects positive on any test were recalled for triage VIA, biopsies, and immediate cryotherapy. Tests were compared using McNemar test., Results: Two thousand forty-nine patients have complete results. Mean age of the patients was 39.2 years; 7.7% presented with ≥atypical squamous cells of uncertain significance (ASCUS), 1.8% ≥low-grade squamous intraepithelial neoplasia, and 0.5% ≥high-grade squamous intraepithelial neoplasia. Two percent of patients had ≥CIN2, and 0.78% had ≥CIN3 (including 2 with invasive disease). The sensitivity of ThinPrep (>ASCUS), HC2, and AHPV for >CIN3 for direct endocervical collection was 87.5%, 100%, and 100%, respectively. The specificity of ThinPrep (>ASCUS), HC2, and AHPV for >CIN3 was 94.1%, 92.2%, and 93.5%, respectively. Specificities of HC2 and AHPV differed significantly. The overall percentage of agreement among HPV assays (HC2 vs APTIMA) is 97%. Four hundred sixty-nine women returned for VIA. Two hundred ninety-one women were treated with cryotherapy., Conclusions: The specificity of the APTIMA assay along with high sensitivity is an advantage for primary screening. Follow-up evaluation will be important to determine the true impact of potential undertreatment in the screening algorithm. Self-sampling applications are explored.

The aim of this work was to characterize the thermal inactivation parameters of recombinant proteins related to the biosynthesis of virgin olive oil (VOO) volatile compounds through the lipoxygenase (LOX) pathway. Three purified LOX isoforms (Oep2LOX1, Oep1LOX2, and Oep2LOX2) and a hydroperoxide lyase (HPL) protein (OepHPL) were studied. According to their thermal inactivation parameters, recombinant Oep1LOX2 and Oep2LOX2 could be identified as the two LOX isoforms active in olive fruit crude preparations responsible for the synthesis of 13-hydroperoxides, the main substrates for the synthesis of VOO volatile compounds. Recombinant Oep2LOX1 displayed a low thermal stability, which suggests a weak actuation during the oil extraction process considering the current thermal conditions of this industrial process. In addition, recombinant OepHPL could be identified as the HPL activity in crude preparations. The thermal stability was the highest among the recombinant proteins studied, which suggests that HPL activity is not a limiting factor for the synthesis of VOO volatile compounds.

The aim of this work was to determine whether the lipoxygenase (LOX) activity is a limiting factor for the biosynthesis of virgin olive oil (VOO) volatile compounds during the oil extraction process. For this purpose, LOX activity load was modified during this process using exogenous LOX activity and specific LOX inhibitors on olive cultivars producing oils with different volatile profiles (Arbequina and Picual). Experimental data suggest that LOX activity is a limiting factor for the synthesis of the oil volatile fraction, this limitation being significantly higher in Picual cultivar than in Arbequina, in line with the lowest content of volatile compounds in the oils obtained from the former. Moreover, there is evidence that this limitation of LOX activity takes place mostly during the milling step in the process of olive oil extraction.

Background: Laparoscopic surgery demands of surgeons special skills acquired only through practice. Laparoscopic training systems traditionally have an optical system that, once positioned, remains fixed and cannot refresh the perspective unless the task is interrupted and the camera repositioned. During a surgery, the visual perspective changes constantly to relocate the surgical target. This difference is a limitation for any novice surgeon. This report proposes the use of a mechatronic system that allows the trainee to handle optics dynamically during training in the cutting task and thereby adapt to dynamic relocation of the surgical target., Methods: The study was conducted in two phases. The first phase involved using fixed optics to cut a circle drawn on a piece of cloth. The second phase involved the same cutting task but with the visual perspective changed dynamically by the user via a mechatronic assistant., Results: The data show that by adapting to dynamic optics, medical trainees can quickly and easily handle and locate the task with real-time changes in visual perspective and can also improve task quality. A significant statistical difference was found between the two methods performed (p < 0.0025). Variance analysis also was applied to the mean values of the scores achieved by both groups (p < 0.0001)., Conclusions: A new laparoscopic training method has been developed. It applies real-time dynamic optics that trainees assist by means of a mechatronic device harnessed to their body. This new training tool allows resident trainees to adapt quickly to the work environment of dynamic optics and thus enter the surgical scenario more rapidly and confidently after mastering the visual-spatial aspect of the laparoscopic approach.

A full-length cDNA clone (OepHPL) coding for hydroperoxide lyase was isolated from olive fruit ( Olea europaea cv. Picual). The deduced amino acid sequence shows significant similarity to known plant hydroperoxide lyases and contains a N-terminal sequence that displays structural features of a chloroplast transit peptide. Genomic Southern blot analysis indicates that at least one copy of OepHPL is present in the olive genome. The recombinant hydroperoxide lyase was specific for 13-hydroperoxide derivatives of linolenic and linoleic acids but did not use 9-hydroperoxy isomers as substrates. Analyses of reaction products revealed that this enzyme produces primarily (Z)-hex-3-enal, which partially isomerizes to (E)-hex-2-enal, from 13-hydroperoxylinolenic acid and hexanal from 13-hydroperoxylinoleic acid. Expression levels were measured in different tissues of Picual and Arbequina varieties, including mesocarp and seed during development and ripening of olive fruits. The involvement of this olive hydroperoxide lyase gene in the biosynthesis of virgin olive oil aroma compounds is discussed.

The endoplasmic reticulum (ER) is where the major histocompatibility complex (MHC) class I molecules are loaded with epitopes to cause an immune cellular response. Most of the protein antigens are degraded in the cytoplasm to amino acids and few epitopes reach the ER. Antigen targeting of this organelle by Calreticulin (CRT) fusion avoids this degradation and enhances the immune response. We constructed a recombinant adenovirus to express the E7 antigen with an ER-targeting signal peptide (SP) plus an ER retention signal (KDEL sequence). In cell-culture experiments we demonstrated that this new E7 antigen, SP-E7-KDEL, targeted the ER. Infection of mice with this recombinant adenovirus that expresses SP-E7-KDEL showed interferon induction and tumour-protection response, similar to that provided by an adenovirus expressing the E7 antigen fused to CRT. This work demonstrated that just by adding a SP and the KDEL sequence, antigens can be targeted and retained in the ER with a consequent enhancement of immune response and tumour protection. These results will have significant clinical applications.

Background: The fine tuning of two features of the bacterial regulatory machinery have been known to contribute to the diversity of gene expression within the same regulon: the sequence of Transcription Factor (TF) binding sites, and their location with respect to promoters. While variations of binding sequences modulate the strength of the interaction between the TF and its binding sites, the distance between binding sites and promoters alter the interaction between the TF and the RNA polymerase (RNAP)., Results: In this paper we estimated the dissociation constants (K(d)) of several E. coli TFs in their interaction with variants of their binding sequences from the scores resulting from aligning them to Positional Weight Matrices. A correlation coefficient of 0.78 was obtained when pooling together sites for different TFs. The theoretically estimated K(d) values were then used, together with the dissociation constants of the RNAP-promoter interaction to analyze activated and repressed promoters. The strength of repressor sites -- i.e., the strength of the interaction between TFs and their binding sites -- is slightly higher than that of activated sites. We explored how different factors such as the variation of binding sequences, the occurrence of more than one binding site, or different RNAP concentrations may influence the promoters' response to the variations of TF concentrations. We found that the occurrence of several regulatory sites bound by the same TF close to a promoter -- if they are bound by the TF in an independent manner -- changes the effect of TF concentrations on promoter occupancy, with respect to individual sites. We also found that the occupancy of a promoter will never be more than half if the RNAP concentration-to-K(p) ratio is 1 and the promoter is subject to repression; or less than half if the promoter is subject to activation. If the ratio falls to 0.1, the upper limit of occupancy probability for repressed drops below 10%; a descent of the limits occurs also for activated promoters., Conclusion: The number of regulatory sites may thus act as a versatility-producing device, in addition to serving as a source of robustness of the transcription machinery. Furthermore, our results show that the effects of TF concentration fluctuations on promoter occupancy are constrained by RNAP concentrations.

An increased methadone enantiomer ratio (R/S) was associated to both nevirapine (179%, n=5) and efavirenz (36%, n=9) treatments when compared with that of controls (n=52). Additionally, in four follow-up patients, both R- and S-methadone normalized concentrations decreased (19%-93%) while R/S increased (22%-314%) following nevirapine/efavirenz treatment. R/S decreased (42%) after non-compliance with efavirenz treatment. Therefore, the methadone-maintenance-treatment outcome should be evaluated when patients are treated with drugs which are supposed to induce CYP3A4 and CYP2B6 isoforms.

Background: The specific recognition of genomic cis-regulatory elements by transcription factors (TFs) plays an essential role in the regulation of coordinated gene expression. Studying the mechanisms determining binding specificity in protein-DNA interactions is thus an important goal. Most current approaches for modeling TF specific recognition rely on the knowledge of large sets of cognate target sites and consider only the information contained in their primary sequence., Results: Here we describe a structure-based methodology for predicting sequence motifs starting from the coordinates of a TF-DNA complex. Our algorithm combines information regarding the direct and indirect readout of DNA into an atomistic statistical model, which is used to estimate the interaction potential. We first measure the ability of our method to correctly estimate the binding specificities of eight prokaryotic and eukaryotic TFs that belong to different structural superfamilies. Secondly, the method is applied to two homology models, finding that sampling of interface side-chain rotamers remarkably improves the results. Thirdly, the algorithm is compared with a reference structural method based on contact counts, obtaining comparable predictions for the experimental complexes and more accurate sequence motifs for the homology models., Conclusion: Our results demonstrate that atomic-detail structural information can be feasibly used to predict TF binding sites. The computational method presented here is universal and might be applied to other systems involving protein-DNA recognition.