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Background and purpose Δ9 -Tetrahydrocannabinolic acid (Δ9 -THCA-A), the precursor of Δ9 -THC, is a non-psychotropic phytocannabinoid that shows PPARγ agonist activity. Here, we investigated the ability of Δ9 -THCA-A to modulate the classic cannabinoid CB1 and CB2 receptors and evaluated its anti-arthritis activity in vitro and in vivo. Experimental approach Cannabinoid receptors binding and intrinsic activity, as well as their downstream signalling, were analysed in vitro and in silico. The anti-arthritis properties of Δ9 -THCA-A were studied in human chondrocytes and in the murine model of collagen-induced arthritis (CIA). Plasma disease biomarkers were identified by LC-MS/MS based on proteomic and elisa assays. Key results Functional and docking analyses showed that Δ9 -THCA-A can act as an orthosteric CB1 receptor agonist and also as a positive allosteric modulator in the presence of CP-55,940. Also, Δ9 -THCA-A seemed to be an inverse agonist for CB2 receptors. In vivo, Δ9 -THCA-A reduced arthritis in CIA mice, preventing the infiltration of inflammatory cells, synovium hyperplasia, and cartilage damage. Furthermore, Δ9 -THCA-A inhibited expression of inflammatory and catabolic genes on knee joints. The anti-arthritic effect of Δ9 -THCA-A was blocked by either SR141716 or T0070907. Analysis of plasma biomarkers, and determination of cytokines and anti-collagen antibodies confirmed that Δ9 -THCA-A mediated its activity mainly through PPARγ and CB1 receptor pathways. Conclusion and implications Δ9 -THCA-A modulates CB1 receptors through the orthosteric and allosteric binding sites. In addition, Δ9 -THCA-A exerts anti-arthritis activity through CB1 receptors and PPARγ pathways, highlighting its potential for the treatment of chronic inflammatory diseases such as rheumatoid arthritis.

We would like to thank Irene Martín Brull, Josep A. Ribera i Torró and Nuria García Díaz for their help in the field. This study was funded by the Spanish Ministry of Economy and Finance (grant CGL2016-79950- R) and the Junta de Andalucia I+D+i program (grant P18-RTJ- 4704)., All data are available from the Dryad Digital Repository https://doi. org/10.5061/dryad.mcvdn ck29 (Rubio de Casas et al., 2021)., The peer review history for this article is available at https://publons.com/publon/10.1111/1365-2745.13831., 1. Cultivated plant species often naturalize and enter wild communities in a process known as feralization. To successfully feralize, crops must overcome ecological barriers and may undergo selection on certain traits, diverging phenotypically and genetically from their crop ancestors. In spite of the agronomic and ecological relevance of crop feralization, the eco-evolutionary dynamics driving it remain understudied. 2. In this paper, we evaluated phenotypic and genotypic differentiation in fruit and seed traits during the naturalization of the almond tree (Prunus dulcis (Mill.) D.A. Webb) in SE Iberia and evaluated the potential role of natural selection in this process. To do so, we investigated the patterns of genetic divergence between cultivated and feral populations using functional (the cyanogenesis Sk gene) and neutral (17 SSR loci) markers and analysed morphological and biochemical traits in kernels of 342 individuals from 15 cultivated and 24 feral populations. 3. We detected very little genetic differentiation in neutral markers between cultivated and feral populations. The majority of the observed genetic variation was due to differences within each type. Conversely, the recessive allele sk responsible for seed toxicity was significantly more frequent in feral populations. Phenotypic differentiation between cultivated and naturalized almond populations was also significant. Feral almond kernels were smaller and lighter, had denser and more resistant shells (endocarps) and more toxic seeds. Selection analyses indicated that these genetic and phenotypic patterns might be driven by directional selection on fruit and seed traits, potentially linked to defence against predation. 4. Synthesis. Our findings indicate that almond naturalization is consistent with strong directional selection on fruits and seeds, leading to smaller and more toxic seeds encased in harder endocarps. Accordingly, we propose that feralization of this crop is, at least to some degree, driven by adaptive evolution of dispersal and recruitment traits., Secretaría de Estado de Investigación, Desarrollo e Innovación, Grant/Award Number: CGL2016-79950- R, Junta de Andalucia I+D+i program (grant P18-RTJ- 4704)

Using social tables, this article provides new data on inequality in Germany and Britain on an annual basis for the first half of the twentieth century. Inequality trends in these two countries tended to follow opposite patterns. The decline in inequality in Germany was interrupted during the First World War and the Nazi period, while in Britain the reversal took place between the end of the First World War and the Great Depression. Results show that the drop in inequality during the twentieth century in Europe did not follow secular trends, thus supporting the notion of inequality cycles.

Background: The effect of leaf removal on postharvest performance of dehydrated grapes has been poorly analyzed. The aim of this study was to evaluate the effect of leaf removal at veraison stage on the metabolites of fresh and dehydrated grapes of two indigenous Cypriot cultivars ('Mavro' and 'Xynisteri'), which are destined for the production of 'Commandaria', a protected designation of origin (PDO) premium sweet wine.; Results: Leaf removal led to a reduction of soluble solids, titratable acidity, aroma potential and most of the phenolic groups in the musts of both cultivars. Dehydration led to a significant increase in all of these parameters in both cultivars, being more pronounced in cv. 'Mavro'. Interestingly, leaf removal indicated differential response in the dehydrated product of the cultivars examined; liquid chromatography (LC-DAD-qTOF-MS) data showed a significant decrease in phenolic compounds in 'Xynisteri' must (from 66.73 to 44.15 mg L-1 ), while 'Mavro' must registered similar values of phenolic compounds (from 94.78 to 96.72 mg L-1 ), but with a different distribution among phenolic groups. Intriguingly, flavonols and flavan-3-ols that present significant health-promoting properties showed higher concentrations in the must from dehydrated 'Mavro' grapes that were subjected to preharvest leaf removal.; Conclusions: Leaf removal at veraison stage followed by sun drying differentially affected the chemical composition of the examined cultivars. This preharvest application followed by postharvest dehydration was beneficial for cv. 'Mavro', while this was not the case for cv. 'Xynisteri'. The effects of leaf removal in other developmental stages (i.e. pre/post bloom stage) need to be dissected with the ultimate goal of providing an end product with high bioactive content. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

Background and purpose The endocannabinoid system and PPARγ are important targets for the development of novel compounds against fibrotic diseases such as systemic sclerosis (SSc), also called scleroderma. The aim of this study was to characterize VCE-004.3, a novel cannabidiol derivative, and study its anti-inflammatory and anti-fibrotic activities. Experimental approach The binding of VCE-004.3 to CB1 and CB2 receptors and PPARγ and its effect on their functional activities were studied in vitro and in silico. Anti-fibrotic effects of VCE-004.3 were investigated in NIH-3T3 fibroblasts and human dermal fibroblasts. To assess its anti-inflammatory and anti-fibrotic efficacy in vivo, we used two complementary models of bleomycin-induced fibrosis. Its effect on ERK1/2 phosphorylation induced by IgG from SSc patients and PDGF was also investigated. Key results VCE-004.3 bound to and activated PPARγ and CB2 receptors and antagonized CB1 receptors. VCE-004.3 bound to an alternative site at the PPARγ ligand binding pocket. VCE-004.3 inhibited collagen gene transcription and synthesis and prevented TGFβ-induced fibroblast migration and differentiation to myofibroblasts. It prevented skin fibrosis, myofibroblast differentiation and ERK1/2 phosphorylation in bleomycin-induced skin fibrosis. Furthermore, it reduced mast cell degranulation, macrophage activation, T-lymphocyte infiltration, and the expression of inflammatory and profibrotic factors. Topical application of VCE-004.3 also alleviated skin fibrosis. Finally, VCE-004.3 inhibited PDGF-BB- and SSc IgG-induced ERK1/2 activation in fibroblasts. Conclusions and implications VCE-004.3 is a novel semisynthetic cannabidiol derivative that behaves as a dual PPARγ/CB2 agonist and CB1 receptor modulator that could be considered for the development of novel therapies against different forms of scleroderma.

Background: Onset during old age has been reported in upto 10% of total cases of inflammatory bowel disease (IBD). Aim: To evaluate phenotypic characteristics and the use of therapeutic resources in patients with elderly onset IBD. Methods: Case-control study including all those patients diagnosed with IBD over the age of 60 years since 2000 who were followed-up for >12 months, identified from the IBD databases. Elderly onset cases were compared with IBD patients aged 18 to 40 years at diagnosis, matched by year of diagnosis, gender and type of IBD (adult-onset). Results: One thousand three hundred and seventy-four elderly onset and 1374 adult-onset cases were included (62% ulcerative colitis (UC), 38% Crohn's disease (CD)). Among UC patients, elderly onset cases had a lower proportion of extensive disease (33% vs 39%; P < 0.0001). In CD, elderly onset cases showed an increased rate of stenosing pattern (24% vs 13%; P < 0.0001) and exclusive colonic location (28% vs 16%; P < 0.0001), whereas penetrating pattern (12% vs 19%; P < 0.0001) was significantly less frequent. Regarding the use of therapeutic resources, there was a significantly lower use of corticosteroids (P < 0.0001), immunosuppressants (P < 0.0001) and anti-TNFs agents (P < 0.0001) in elderly onset cases. Regarding surgery, we found a significantly higher surgery rate among elderly onset UC cases (8.3% vs 5.1%; P < 0.009). Finally, elderly onset cases were characterised by a higher rate of hospitalisations (66% vs 49%; P < 0.0001) and neoplasms (14% vs 0.5%; P < 0.0001). Conclusions: Elderly onset IBD shows specific characteristics and they are managed differently, with a lower use of immunosuppressants and a higher rate of surgery in UC.

Summary The current economic crisis has increased the interest in analyzing the efficiency of health care systems, as their funding is a very important part of the budgets for different countries. In this work determines the efficiency in the health services in European countries applying data envelopment analysis. In addition, the combined application of data envelopment analysis methods and ACP can provide an evaluation of the efficiency with respect to differently oriented productive health systems in the different countries. The results show that models with a lower level of efficiency are those whose input is beds, followed by the models whose input is physicians. Finally, we apply the AD to select a few simple indicators that facilitate control of the level of operational efficiency of a health system.

Pseudomonas putida strains are generally recognized as solvent tolerant, exhibiting varied sensitivity to organic solvents. Pan-genome analysis has revealed that 30% of genes belong to the core-genome of Pseudomonas. Accessory and unique genes confer high degree of adaptability and capabilities for the degradation and synthesis of a wide range of chemicals. For the use of these microbes in bioremediation and biocatalysis, it is critical to understand the mechanisms underlying these phenotypic differences. In this study, RNA-seq analysis compared the short- and long-term responses of the toluene-sensitive KT2440 strain and the highly tolerant DOT-T1E strain. The sensitive strain activates a larger number of genes in a higher magnitude than DOT-T1E. This is expected because KT2440 bears one toluene tolerant pump, while DOT-T1E encodes three of these pumps. Both strains activate membrane modifications to reduce toluene membrane permeability. The KT2440 strain activates the TCA cycle to generate energy, while avoiding energy-intensive processes such as flagellar biosynthesis. This suggests that KT2440 responds to toluene by focusing on survival mechanisms. The DOT-T1E strain activates toluene degradation pathways, using toluene as source of energy. Among the unique genes encoded by DOT-T1E is a 70 kb island composed of genes of unknown function induced in response to toluene.

Objective MircroRNAs (miR) are small, noncoding RNA molecules of 18-25 nucleotides. Their dysregulation has been widely studied in many human tumours including differentiated thyroid cancer (DTC). miRs more frequently associated with these kinds of tumours are miR-146, miR-221 and miR-222. Our objective was to assess the relationship among circulating miR levels and the evolution and outcomes of disease. Design We analysed a sample of 60 patients with DTC assigning them to one of three groups according to the dynamic scale of risk (excellent response, incomplete biochemical response and incomplete structural response). Patients and measurements At study inclusion, we determined thyroid-stimulating hormone, thyroxine, thyroglobulin, antithyroglobulin antibodies and plasma levels of miR-146, miR-221 and miR-222. Results Male sex and advanced age at diagnosis were associated with the worst disease progression. miR-222 was twofold to threefold higher in tall cell papillary carcinomas (P = 0.038). miR-146 (P = 0.016) and miR-221 (P = 0.050) had a positive correlation with thyroglobulin at the time of sampling. In regression analysis, miR-146 (P = 0.006), miR-221 (P = 0.004) and miR-222 (P = 0.007) predicted more than 70% of the variation in thyroglobulin levels at the time of sampling. Conclusions Elevated miR-222 and miR-146 levels are associated with poorer outcomes of the disease and may have a prognostic value in the management and follow-up of DTC.

Caffeine has cognitive-enhancing properties with effects on learning and memory, concentration, arousal and mood. These effects imply changes at circuital and synaptic level, but the mechanism by which caffeine modifies synaptic plasticity remains elusive. Here we report that caffeine, at concentrations representing moderate to high levels of consumption in humans, induces an NMDA receptor-independent form of LTP (CAFLTP) in the CA1 region of the hippocampus by promoting calcium-dependent secretion of BDNF, which subsequently activates TrkB-mediated signaling required for the expression of CAFLTP. Our data include the novel observation that insulin receptor substrate 2 (IRS2) is phosphorylated during induction of CAFLTP, a process that requires cytosolic free Ca2+. Consistent with the involvement of IRS2 signals in caffeine-mediated synaptic plasticity, phosphorylation of Akt (Ser473) in response to LTP induction is defective in Irs2−/− mice, demonstrating that these plasticity changes are associated with downstream targets of the phosphoinositide 3-kinase (PI3K) pathway. These findings indicate that TrkB-IRS2 signals are essential for activation of PI3K during the induction of LTP by caffeine.

Free and bound phenolic and other polar compounds in mango edible fraction and its by-products (peel, seed, and seed husk) have been determined by HPLC-DAD-ESI-qTOF-MS. This analytical technique has demonstrated to be a valuable platform for the identification and quantification of these compounds in mango. In fact, UV-Vis and mass spectra data allowed the determination of 91 free compounds and 13 bound (cell wall linked) compounds taking into account the four fractions of mango. To our knowledge, this is the first time that mango seed husk has been studied regarding its phenolic compounds. The method proposed showed LODs between 0.006 and 0.85 μg/mL and accuracy ranged from 94.8 and 100.7%. Mango peel presented the highest concentration of free polar compounds followed by seed, pulp, and seed husk. It is also important to highlight that bound phenolic compounds had never been determined in mango pulp, seed, and seed husk before. Furthermore, ellagic acid was the most abundant bound compound in the four mango fractions analyzed. These results show that mango pulp and its by-products are a good source of phenolic and other polar compounds. In particular, mango seed contains a high total concentration of ellagic acid (650 mg/100 g dry weight).

The aim of our study was to determine the frequency and type of associated congenital anomalies in patients with isolated gastroschisis born at the Dr. Juan I. Menchaca Civil Hospital of Guadalajara (Guadalajara, México), and to explore its possible association with the included outcome variables. One hundred-eight cases with isolated gastroschisis were reviewed from 2009 to 2014. The occurrence of intestinal and extraintestinal associated anomalies (either secondary or primary) was prospectively assessed. The type of gastroschisis, length of hospital stay (LOS), and in-hospital mortality were outcome variables for statistical analysis. Of infants with gastroschisis, 52 (48.1%) had one or more associated anomalies (AA), with increased odds in males (OR = 2.3, 95%CI: 1.1-5.0). AA classified, as secondary and primary were present in 34.3 and 5.6% of patients, respectively. Of secondary AA, 25.9% were intestinal anomalies, and 17.6% were extraintestinal. Primary AA were congenital heart disease (n = 3), meningomyelocele, and hydrocephaly and amniotic band sequence in one instance, respectively. Multivariate logistic regression showed that secondary AA (both intestinal and extraintestinal) were associated with complex gastroschisis, prolonged LOS, and in-hospital death, whereas primary AA were not related to a worse outcome. Our results highlight the pathogenic importance of properly investigating and categorizing the presence of others secondary or primary AA when diagnosis of gastroschisis is made.

BACKGROUND The present work deals with the evaluation of the effect of three different commercial starter cultures (Chr. Hansen, Horsholm, Denmark) on the volatile compound profile and sensory properties, as well as some important physicochemical parameters, of dry-fermented foal sausages at the end of ripening in order to select the most suitable starter culture for this elaboration. The sausage batches were named as follows: CO (non-inoculated control), FS (Lactobacillus sakei + Staphylococcus carnosus), SM (L. sakei + S. carnosus + Staphylococcus xylosus + Pediococcus pentosaceus + Debaryomyces hansenii) and TR (L. sakei + S. carnosus +S. xylosus). RESULTS The pH values differed significantly among batches, with the highest values corresponding to CO followed by TR, SM and FS. The highest amounts of volatile compounds were found in FS batch. Hexanal was the most abundant compound, especially in FS and SM batches. These batches also showed higher levels of compounds derived from carbohydrate fermentation and amino acid catabolism. Sensory results showed that acid taste was significantly lower in CO batch than in inoculated batches. CONCLUSION According to most parameters, batches inoculated with FS and SM starters showed marked acidity compared with TR and CO batches, as expected from the manufacturer's indications. Therefore the most suitable starter culture for use in the manufacture of foal sausages in Mediterranean countries such as Spain with a preference for low-acidity products was found to be TR culture. © 2015 Society of Chemical Industry

Summary Multi-drug resistant bacteria are a major threat to humanity, especially because the current battery of known antibiotics is not sufficient to combat infections produced by these microbes. Therefore, the study of how current antibiotics act and how bacteria defend themselves against antibiotics is of critical importance. Pseudomonas putida DOT-T1E exhibits an impressive array of RND efflux pumps, which confer this microorganism high resistance to organic solvents and antibiotics that would kill most other microorganisms. We have chosen DOT-T1E as a model microbe to study the microbial responses to a wide battery of antibiotics (chloramphenicol, rifampicin, tetracycline, ciprofloxacin, ampicillin, kanamycin, spectinomycin and gentamicin). Ribonucleic acid sequencing (RNA)-seq analyses revealed that each antibiotic provokes a unique transcriptional response profile in DOT-T1E. While many of the genes identified were related to known antibiotic targets, others were unrelated or encoded hypothetical proteins. These results indicate that our knowledge of antibiotic resistance mechanisms is still partial. We also identified 138 new small RNAs (sRNAs) in DOT-T1E, dramatically adding to the 16 that have been previously described. Importantly, our results reveal that a correlation exists between the expression of messenger RNA and sRNA, indicating that some of these sRNAs are likely involved in fine tuning the expression of antibiotic resistance genes. Taken together, these findings open new frontiers in the fight against multi-drug resistant bacteria and point to the potential use of sRNAs as novel antimicrobial targets.

Background Biomarkers of Alzheimer’s disease (AD) pathology and progression have now been identified across various modalities. The aims of the two studies presented (ANM-MMB and EMIF-AD biomarker discovery) are to discover and replicate previously identified biomarkers of disease pathology and progression, and moreover to determine whether a multimodal biomarker signature may add value in comparison to a biomarker of a single modality. Methods The ANM-MMB cohort is comprised of 718 AD, MCI converters and non-converters, and control subjects selected from the AddNeuroMed, Alzheimer’s research trust and Dementia case register cohorts. Cognitive measures, serum and urine metabolomics, structural MRI, genomics, whole blood transcriptomics and plasma proteomics data was available. The EMIF-AD biomarker discovery cohort consists of 1221 AD, MCI and control subjects, selected from the EMIF catalogue. All subjects had existing amyloid measures (CSF Aβ or amyloid-PET), structural MRI and clinical data, and furthermore plasma proteomics (targeted and untargeted), CSF proteomics (targeted), metabolomics, genomics and epigenetics data were generated. For both studies univariate and multivariate statistics were utilised to identify candidate biomarkers of AD pathology (neurodegeneration and/or brain amyloid burden), rates of cognitive decline, and MCI progression to dementia. pQTL-eQTL-mQTL analyses, network/pathway analysis, and multimodal classifiers were employed to detect multimodal signatures. Results Initial analyses indicate that in the ANM-MMB study a serum and urine derived 15 metabolite classifier predicts MCI progression to AD with 72% accuracy, and the biological significance of the metabolites included in the biomarker panel was identified. Further analyses will examine whether a multimodal classifier is able to predict with even greater accuracy. We will then seek to replicate this in the EMIF-AD biomarker discovery study. Further analyses will also examine single and multimodal biomarker classifiers of other endophenotypes. Conclusions These two studies could be used to identify novel and replicate previously identified single modality biomarker findings. Furthermore the impact of combining the additional modalities with these findings will be discussed. Computational and technical challenges encountered and the bioinformatics pipeline devised in the multimodal analysis of the ANM-MMB cohort will be used to inform the analysis pipeline of the EMIF-AD biomarker discovery study as a replication.

This work aims to contribute to the discussion about the suitability of bond paths and bond-critical points as indicators of chemical bonding defined within the theoretical framework of the quantum theory of atoms in molecules. For this purpose, we consider the temporal evolution of the molecular structure of [Fe{C(CH2 )3 }(CO)3 ] throughout Born-Oppenheimer molecular dynamics (BOMD), which illustrates the changing behaviour of the molecular graph (MG) of an electronic system. Several MGs with significant lifespans are observed across the BOMD simulations. The bond paths between the trimethylenemethane and the metallic core are uninterruptedly formed and broken. This situation is reminiscent of a "hopping" ligand over the iron atom. The molecular graph wherein the bonding between trimethylenemethane and the iron atom takes place only by means of the tertiary carbon atom has the longest lifespan of all the considered structures, which is consistent with the MG found by X-ray diffraction experiments and quantum chemical calculations. In contrast, the η(4) complex predicted by molecular-orbital theory has an extremely brief lifetime. The lifespan of different molecular structures is related to bond descriptors on the basis of the topology of the electron density such as the ellipticities at the FeCH2 bond-critical points and electron delocalisation indices. This work also proposes the concept of a dynamic molecular graph composed of the different structures found throughout the BOMD trajectories in analogy to a resonance hybrid of Lewis structures. It is our hope that the notion of dynamic molecular graphs will prove useful in the discussion of electronic systems, in particular for those in which analysis on the basis of static structures leads to controversial conclusions.

Background and PurposeSpinocerebellar ataxias (SCAs) are a family of chronic progressive neurodegenerative diseases, clinically and genetically heterogeneous, characterized by loss of balance and motor coordination due to degeneration of the cerebellum and its afferent and efferent connections. Unlike other motor disorders, the possible role of changes in the endocannabinoid system in the pathogenesis of SCAs has not been investigated. Experimental ApproachThe status of cannabinoid receptor type 1 (CB1) and cannabinoid receptor type 2 (CB2) receptors in the post-mortem cerebellum of SCA patients and controls was investigated using immunohistochemical procedures. Key ResultsImmunoreactivity for the CB1 receptor, and also for the CB2 receptor, was found in the granular layer, Purkinje cells, neurons of the dentate nucleus and areas of white matter in the cerebellum of SCA patients at levels notably higher than controls. Double-labelling procedures demonstrated co-localization of CB1 and, in particular, CB2 receptors with calbindin, supporting the presence of these receptors in Purkinje neurons. Both receptors also co-localized with Iba-1 and glial fibrillary acidic protein in the granular layer and white matter areas, indicating that they are present in microglia and astrocytes respectively. Conclusions and ImplicationsOur results demonstrate that CB1 and CB2 receptor levels are significantly altered in the cerebellum of SCA patients. Their identification in Purkinje neurons, which are the main cells affected in SCAs, as well as the changes they experienced, suggest that alterations in endocannabinoid receptors may be related to the pathogenesis of SCAs. Therefore, the endocannabinoid system could provide potential therapeutic targets for the treatment of SCAs and its progression. Linked ArticlesThis article is part of a themed section on Cannabinoids 2013. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-6

This study measures the total factor productivity ( TFP) of 15 OECD countries in the period 1989-2004 and disaggregates it into its two components: technical change and efficiency change. Subsequently, two theoretical models are estimated to evaluate the effects that technology and institutions have on technical change and on efficiency change. The results show that technology, proxied by domestic R&D stocks or by patents, stimulated the variation in technical change and in efficiency change. In contrast, the reduction of entry barriers and of the relative weight of publicly owned companies, or deregulation in the telecommunications sector, drove technical change but, at the same time, slowed efficiency change.

BACKGROUND Bovine milk contains hundreds of diverse components, including proteins, peptides, amino acids, lipids, lactose, vitamins and minerals. Specifically, the lipid composition is influenced by different variables such as breed, feed and technological process. In this study the fatty acid and phospholipid compositions of different samples of butter and its by-products from the Parmigiano Reggiano cheese area, produced by industrial and traditional churning processes, were determined. RESULTS The fatty acid composition of samples manufactured by the traditional method showed higher levels of monounsaturated and polyunsaturated fatty acids compared with industrial samples. In particular, the contents of n-3 fatty acids and conjugated linoleic acids were higher in samples produced by the traditional method than in samples produced industrially. Sample phospholipid composition also varied between the two technological processes. Phosphatidylethanolamine was the major phospholipid in cream, butter and buttermilk samples obtained by the industrial process as well as in cream and buttermilk samples from the traditional process, while phosphatidylcholine was the major phospholipid in traditionally produced butter. This result may be explained by the different churning processes causing different types of membrane disruption. Generally, samples produced traditionally had higher contents of total phospholipids; in particular, butter produced by the traditional method had a total phospholipid content 33% higher than that of industrially produced butter. CONCLUSION The samples studied represent the two types of products present in the Parmigiano Reggiano cheese area, where the industrial churning process is widespread compared with the traditional processing of Reggiana cow's milk. This is because Reggiana cow's milk production is lower than that of other breeds and the traditional churning process is time-consuming and economically disadvantageous. However, its products have been demonstrated to contain more bioactive lipids compared with products obtained from other breeds and by the industrial process. © 2013 Society of Chemical Industry

Bacterial small regulatory RNAs (sRNAs) function in post-transcriptional control of gene expression and control a variety of processes including metabolic reactions, stress responses and pathogenesis in response to environmental signals. A variety of approaches have been used previously to identify 44 sRNAs in the opportunistic human pathogen Pseudomonas aeruginosa. In this work, RNA sequencing (RNA-seq) is used to identify novel transcripts in P.aeruginosa involving a combination of three different sequencing libraries. Almost all known sRNAs and over 500 novel intergenic sRNAs are identified with this approach. Although the use of three libraries increased the number of novel transcripts identified, there were significant differences in the subset of transcripts detected in each library, underscoring the importance of library preparation strategy and relative sRNA abundance for successful sRNA detection. Nearly 90% of the novel sRNAs have no orthologous bacterial sequences outside of P.aeruginosa, supporting a limited degree of sequence conservation and rapid evolution of sRNAs at the species level. We anticipate that the data will be useful for the study of regulatory sRNAs in bacteria and that the approach described here may be applied to identify sRNAs in any bacterium under different growth and stress conditions.

Surface modification of a microporous polypropylene (PP) membrane was performed by graft polymerization of acrylated epoxidized soybean oil (AESO) using UV radiation. This is a simple, environmentally friendly, and low cost method. The factors affecting the grafting degree such as monomer concentration and reaction time were studied. The morphological and microstructure changes of the membrane were characterized by scanning electron microscopy (SEM) and information regarding elemental composition was obtained by energy dispersive X-ray spectroscopy (EDS). Surface functionalization was studied by X-ray photoelectron spectroscopy (XPS). Thermal properties were analyzed by differential scanning calorimetry (DSC). The hydrophilicity increment was confirmed by pure water contact angle and swelling measurements. The filtration capability of the modified membrane was analyzed by determining color, turbidity, and chemical oxygen demand (COD) removal from a residual raw water. The results indicated that the contact angle of pure water on the grafted membrane decreased from 90° to 57°. The modified membrane shows filtration capability by removing color in 52% and reducing 95% of turbidity. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Fumaric acid (FA) was produced from Eucalyptus globulus wood by successive steps of hydrothermal processing (to solubilize hemicelluloses and to increase the susceptibility of solids to enzymatic hydrolysis), enzymatic hydrolysis and fermentation with Rhizopus arrhizus DSM 5772. For comparative purposes, additional fermentations were carried out using synthetic media. Single stage fermentation of synthetic media led to a medium containing 11.8 g FA L−1 (YP/S = 0.60 g g−1). Operating in fed batch mode, the fourth stage increased the FA concentration from 19.7 up to 43.6 g L−1 (YP/S = 0.71 g g−1). Hydrolyzate fermentation in a single stage resulted in lower fumaric acid concentration (9.65 g L−1) and yield (0.35 g g−1). Additional fermentations were carried out in media made with hydrolyzates subjected to membrane processing, adsorption or ion exchange. The highest yield (YP/S = 0.44 g g−1) was reached in media made up of ion-exchange treated hydrolyzates and a commercial glucose solution in proportion 85/15 w/w. Copyright © 2011 Society of Chemical Industry

Centrin is a calcium binding protein (CaBP) belonging to the EF-hand superfamily. As with other proteins within this family, centrin is a calcium sensor with multiple biological target proteins. We chose to study Chlamydomonas reinhardtii centrin (Crcen) and its interaction with melittin (MLT) as a model for CaBP complexes due to its amphipathic properties. Our goal was to determine the molecular interactions that lead to centrin-MLT complex formation, their relative stability, and the conformational changes associated with the interaction, when compared to the single components. For this, we determined the thermodynamic parameters that define Crcen-MLT complex formation. Two-dimensional infrared (2D IR) correlation spectroscopy were used to study the amide I', I'*, and side chain bands for (13)C-Crcen, MLT, and the (13)C-Crcen-MLT complex. This approach resulted in the determination of MLT's increased helicity, while centrin was stabilized within the complex. Herein we provide the first complete molecular description of centrin-MLT complex formation and the dissociation process. Also, discussed is the first structure of a CaBP-MLT complex by X-ray crystallography, which shows that MLT has a different binding orientation than previously characterized centrin-bound peptides. Finally, all of the experimental results presented herein are consistent with centrin maintaining an extended conformation while interacting with MLT. The molecular implications of these results are: (1) the recognition of hydrophobic contacts as requirements for initial binding, (2) minimum electrostatic interactions within the C-terminal end of the peptide, and (3) van der Waals interactions within MLTs N-terminal end are required for complex formation.

This work describes the conformational behavior and the activation mechanism of timoprazole and substituted prazoles from the most stable conformation to the sulphenic acid. The stability of the conformers can be explained by the presence of hydrogen bonds, stereoelectronic effect because of the lone pair of sulfur atom and the N…C and N…S interactions. The first step of the Smile rearrangement is a nucleophilic addition to benzimidazole by pyridine moiety, which depends on the difference of the electron population of the atoms involved in the attack. The second step produces sulphenic acid by a concerted reaction where breaking of the S–C bond goes along with a proton migration, and is determined by the electron population of the sulfur atom. Copyright © 2011 John Wiley & Sons, Ltd.

Cannabinoids are promising medicines to slow down disease progression in neurodegenerative disorders including Parkinson's disease (PD) and Huntington's disease (HD), two of the most important disorders affecting the basal ganglia. Two pharmacological profiles have been proposed for cannabinoids being effective in these disorders. On the one hand, cannabinoids like Δ(9) -tetrahydrocannabinol or cannabidiol protect nigral or striatal neurons in experimental models of both disorders, in which oxidative injury is a prominent cytotoxic mechanism. This effect could be exerted, at least in part, through mechanisms independent of CB(1) and CB(2) receptors and involving the control of endogenous antioxidant defences. On the other hand, the activation of CB(2) receptors leads to a slower progression of neurodegeneration in both disorders. This effect would be exerted by limiting the toxicity of microglial cells for neurons and, in particular, by reducing the generation of proinflammatory factors. It is important to mention that CB(2) receptors have been identified in the healthy brain, mainly in glial elements and, to a lesser extent, in certain subpopulations of neurons, and that they are dramatically up-regulated in response to damaging stimuli, which supports the idea that the cannabinoid system behaves as an endogenous neuroprotective system. This CB(2) receptor up-regulation has been found in many neurodegenerative disorders including HD and PD, which supports the beneficial effects found for CB(2) receptor agonists in both disorders. In conclusion, the evidence reported so far supports that those cannabinoids having antioxidant properties and/or capability to activate CB(2) receptors may represent promising therapeutic agents in HD and PD, thus deserving a prompt clinical evaluation.

The behavior of different reverse osmosis membranes, namely, HR98PP, SEPA-MS05, and DESAL-3B, was compared with the one predicted by a solution-diffusion model. This model assumes that the membrane has a homogeneous, nonporous surface layer and uses two main parameters, permeate concentration Cp and volumetric permeate flux Qp, to characterize the membrane system. In order to calculate these parameters, the theoretical values of water and solute permeability constants, Aw and Bs, and the osmotic pressure coefficient, Ψ, were initially determined. Using the software Sigma Plot V 10.0, accurate values of Ψ, Qp, and Cp were only obtained for the DESAL-3B membrane, i.e., the selected solution-diffusion model can be applied to this membrane.

BACKGROUND AND PURPOSE The high predisposition to Torsade de Pointes (TdP) in dogs with chronic AV-block (CAVB) is well documented. The anti-arrhythmic efficacy and mode of action of Ca2+ channel antagonists, flunarizine and verapamil against TdP were investigated. EXPERIMENTAL APPROACH Mongrel dogs with CAVB were selected based on the inducibility of TdP with dofetilide. The effects of flunarizine and verapamil were assessed after TdP and in different experiments to prevent dofetilide-induced TdP. Electrocardiogram and ventricular monophasic action potentials were recorded. Electrophysiological parameters and short-term variability of repolarization (STV) were determined. In vitro, flunarizine and verapamil were added to determine their effect on (i) dofetilide-induced early after depolarizations (EADs) in canine ventricular myocytes (VM); (ii) diastolic Ca2+ sparks in RyR2R4496+/+ mouse myocytes; and (iii) peak and late INa in SCN5A-HEK 293 cells. KEY RESULTS Dofetilide increased STV prior to TdP and in VM prior to EADs. Both flunarizine and verapamil completely suppressed TdP and reversed STV to baseline values. Complete prevention of TdP was achieved with both drugs, accompanied by the prevention of an increase in STV. Suppression of EADs was confirmed after flunarizine. Only flunarizine blocked late INa. Ca2+ sparks were reduced with verapamil. CONCLUSIONS AND IMPLICATIONS Robust anti-arrhythmic efficacy was seen with both Ca2+ channel antagonists. Their divergent electrophysiological actions may be related to different additional effects of the two drugs.

Most CE methods for the analysis of phenols from olive oil use an aqueous electrolyte separation medium, although the importance of NACE is obvious, as this kind of CE seems to be more compatible with the hydrophobic olive oil matrix and could facilitate its direct injection. In the current work we develop a method involving SPE and NACE coupled to ESI-TOF MS. All the CE and ESI-TOF MS parameters were optimized in order to maximize the number of phenolic compounds detected and the sensitivity in their determination. Electrophoretic separation was carried out using a CE buffer system consisting of 25 mM NH(4)OAc/AcH in methanol/ACN (1/1 v/v) at an apparent pH value of 5.0. We studied in depth the effect of the nature and concentration of different electrolytes dissolved in different organic solvents and other experimental and instrumental CE variables. The results were compared with those obtained by CZE (with aqueous buffers) coupled to ESI-TOF MS; both methods offered to the analyst the chance to study phenolic compounds of different families (such as phenolic alcohols, lignans, complex phenols, flavonoids, etc.) from virgin olive oil by injecting methanolic extracts with efficient and fast CE separations. In the case of NACE method, we also studied the direct injection of the investigated matrix introducing a plug of olive oil directly into the capillary.

The immobilization of trypsin onto various commercial silica gels was studied. Silica gels were used directly and characterized by mercuric porosimetry. Agitation rates (100-740 rpm) and particles size (35-75 to 250-500 μm) of silica gels did not affect the trypsin immobilization capacity. The pore size (3 to 15 nm) is a limiting factor of the trypsin adsorption onto the mesopores structure of silica gels. The adsorption of trypsin was determined as a function of their initial concentration and multilayer formed at high trypsin concentration.

BACKGROUND: There is much interest in foods that belong to Protected Designations of Origin (PDO) because the quality and other essential and exclusive characteristics are guaranteed because of a particular geographical environment. We have used a capillary electrophoretic method for the simple, rapid and simultaneous characterization and quantification of the polyphenolic fraction of extra-virgin olive oil from different PDOs (18 compounds in less than 7 min). To demonstrate the usefulness of this method, we have analyzed 16 samples of a Spanish PDO and nine other samples belonging to an Italian PDO (n = 5). RESULTS: In this way, it was possible to compare the phenolic profiles of the oils of different zones of the same PDO, as well as the phenolic profiles of Spanish and Italian extra-virgin olive oils. Univariate statistics were used for differentiating the oils produced in each PDO. Furthermore, the correlations among several of the phenolic compounds present in the extracts of olive oil and its sensorial properties were checked. decarboxylated oleuropein aglycon (DOA) (b) (peak 6) was the compound more related to the bitterness of the oils, Ac Pin (peak number 4) and the unidentified peak with tmig 4.025 min played an important role in the pungent taste of the oils, and these two latter compounds and the peak 13 seemed to be related to the fruitiness of the oils. CONCLUSIONS: The electrophoretic method described in the current work has the ability to detect and quantify simultaneously 18 phenolic compounds (belonging to five different families) in less than 7 min. This technique could improve the characterization of this polar fraction and determine the geographical origin of olive oils or detect possible ‘PDO markers’. In fact, capillary electrophoresis coupled to statistical analysis enabled discrimination among olive oils belonging to two different PDOs considering eight phenolic compounds present in the extracts, and also identification of the phenolic compounds which could have more influence on several sensory attributes such as bitterness, pungency and fruitiness. Copyright © 2009 Society of Chemical Industry

Temperature not only has direct effects on microbial activity, but can also affect activity indirectly by changing the temperature dependency of the community. This would result in communities performing better over time in response to increased temperatures. We have for the first time studied the effect of soil temperature (5-50 degrees C) on the community adaptation of both bacterial (leucine incorporation) and fungal growth (acetate-in-ergosterol incorporation). Growth at different temperatures was estimated after about a month using a short-term assay to avoid confounding the effects of temperature on substrate availability. Before the experiment started, fungal and bacterial growth was optimal around 30 degrees C. Increasing soil temperature above this resulted in an increase in the optimum for bacterial growth, correlated to soil temperature, with parallel shifts in the total response curve. Below the optimum, soil temperature had only minor effects, although lower temperatures selected for communities growing better at the lowest temperature. Fungi were affected in the same way as bacteria, with large shifts in temperature tolerance at soil temperatures above that of optimum for growth. A simplified technique, only comparing growth at two contrasting temperatures, gave similar results as using a complete temperature curve, allowing for large scale measurements also in field situations with small differences in temperature. (Less)

This article focuses on the study of patinas by means of a non-destructive and fast energy dispersive x-ray fluorescence (EDXRF) technique using portable equipment. The term patina is referred here to thin films or coverings (5-500 μm thick) on the surface of stone building materials, characterized by a common yellowish colour and a common presence of calcium carbonate, calcium sulphate, calcium oxalate, calcium phosphate, silicate and iron oxide and/or hydroxide. Some of these patinas are the result of applied past treatments with protective and/or aesthetical purposes. The main aim of this article is to assess the potential of the technique, together with the determination of the elemental composition of the samples and the provision of analytical information leading to the discrimination between different types of patinas. This work proved that the EDXRF spectrometry is a useful technique for qualitative and even semi-quantitative analyses of patinas in situ. It is a reliable method for a first approach to the study of a patina of any building. The results obtained showed that the patina is characterized by the presence and enrichment of elements like Si, P, K, Fe, Ti and Mn. For this reason, they could be considered as characteristics of this kind of material. Copyright © 2008 John Wiley & Sons, Ltd., This study has been financed by project BIA2003-0473 (Spanish Ministry of Education and Science), by a ‘Ramon y Cajal’ contract (MAdB) (Ministry of Education and Science). CVC thanks the CSIC (Spanish Council for Scientific Research) Thematic Network for Historical and Cultural Heritage for a scholarship co-financed by the European Social Found.

cAMP is a powerful second messenger whose known general effector is protein kinase A (PKA). The identification of a cAMP binding protein, Epac, raises the question of its role in Ca(2+) signalling in cardiac myocytes. In this study, we analysed the effects of Epac activation on Ca(2+) handling by using confocal microscopy in isolated adult rat cardiomyocytes. [Ca(2+)](i) transients were evoked by electrical stimulation and Ca(2+) sparks were measured in quiescent myocytes. Epac was selectively activated by the cAMP analogue 8-(4-chlorophenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate (8-CPT). Patch-clamp was used to record the L-type calcium current (I(Ca)), and Western blot to evaluate phosphorylated ryanodine receptor (RyR). [Ca(2+)](i) transients were slightly reduced by 10 microm 8-CPT (F/F(0): decreased from 4.7 +/- 0.5 to 3.8 +/- 0.4, P < 0.05), an effect that was boosted when cells were previously infected with an adenovirus encoding human Epac. I(Ca) was unaltered by Epac activation, so this cannot explain the decreased [Ca(2+)](i) transients. Instead, a decrease in the sarcoplasmic reticulum (SR) Ca(2+) load underlies the decrease in the [Ca(2+)](i) transients. This decrease in the SR Ca(2+) load was provoked by the increase in the SR Ca(2+) leak induced by Epac activation. 8-CPT significantly increased Ca(2+) spark frequency (Ca(2+) sparks s(-1) (100 microm)(-1): from 2.4 +/- 0.6 to 6.9 +/- 1.5, P < 0.01) while reducing their amplitude (F/F(0): 1.8 +/- 0.02 versus 1.6 +/- 0.01, P < 0.001) in a Ca(2+)/calmodulin kinase II (CaMKII)-dependent and PKA-independent manner. Accordingly, we found that Epac increased RyR phosphorylation at the CaMKII site. Altogether, our data reveal a new signalling pathway by which cAMP governs Ca(2+) release and signalling in cardiac myocytes.

Diets in the countries of the Mediterranean basin are characterised by abundant plant foods (fruits, vegetables, breads, nuts, seeds, wine, and olive oil) and include fish and low-fat dairy products. Among the vegetables, tomatoes are a main component of the traditional Mediterranean diet, which has been associated with health protection and longevity. Eating tomatoes has been associated with reduced risks of some types of cancer and other diseases. These beneficial properties appear to be related to the antioxidant content of the fruit, particularly carotenoids (lycopene and beta-carotene), ascorbic acid, and phenols, which may play a role in inhibiting reactions mediated by reactive oxygen species. Due to the importance of antioxidant compounds in tomatoes and tomato products, we present here an overview of current analytical methods (from 2000 until the present date) for determining the different antioxidants. The analytical procedures used to determine individual compounds involve extraction from the sample, analytical separation, and quantification. The choice of analytical method depends on the particular focus of the analysis and the kind of product analysed. High-performance liquid chromatography is the technique of choice for the analysis of tomato antioxidants.

The aim of this study was to compare the performance of the Bishop score and transvaginal ultrasonography to predict successful labor induction, and to estimate the most useful cut-off points for the two methods.The five components of the Bishop score were assessed by digital examination and cervical length was measured by transvaginal ultrasonography in 177 women with a single pregnancy, 36-42 weeks of gestation, and a live fetus in cephalic presentation before induction of labor.Multiple regression analysis showed that the Bishop score, cervical length, and parity provided independent contribution in the prediction of the likelihood of delivering vaginally within 60 h. The only component of the Bishop score that was independently predictive of the probability of vaginal delivery within 60 h was station. The best cut-off points for predicting successful induction using receiver operating characteristic curves were 24 mm for cervical length and 4 for the Bishop score. Cervical length was a better predictor than the Bishop score (sensitivity and specificity of 66 and 77% versus 77 and 56%, respectively). Kaplan-Meier survival curves showed that cervical length was the best discriminator of successful induction.Measurement of cervical length by transvaginal ultrasonography is a better predictor of success in labor induction with both topical prostaglandin and oxytocin.