Quantification of gliadin levels to the picogram level by flow cytometry

Celiac disease is a widely prevalent enteropathy caused by intolerance to gliadin, one of the gluten proteins. We developed two methods for the analysis of gliadin levels. Both methods use flow cytometry and rat antibodies against a 16-residue peptide of gliadin. The peptide is common to the alpha-, beta-, gamma-, and omega-gliadins

The dark halo of the Hydra I galaxy cluster: core, cusp, cosmological? Dynamics of NGC 3311 and its globular cluster system

NGC 3311 is the central cD galaxy of the Hydra I cluster. We use globular clusters around NGC 3311, combined with kinematical data of the galaxy itself, to investigate the dark matter distribution in the central region of Hydra I. Radial velocities of 118 bright globular clusters, based on VLT/VIMOS mask spectroscopy, are used to calculate velocity dispersions which are well defined out to 100 kpc. NGC 3311 is the most distant galaxy for which this kind of study has been performed. We also determine velocity dispersions of the stellar component from long slit spectroscopy out to 20 kpc. Moreover, we present a new photometric model for NGC 3311 in the V-band. We search for a dark halo which in the context of a spherical Jeans model. We also compare the radial velocity distributions of globular clusters and planetary nebulae. The projected stellar velocity dispersion rises from 185 km/s to 350 km/s at a radius of 20 kpc. The globular cluster dispersion rises as well from 500 km/s at 10 kpc to about 800 km/s at 100 kpc, comparable to the velocity dispersion of the cluster galaxies. A dark matter halo with a core reproduces well the velocity dispersions of stars and globular clusters simultaneously under isotropy. The central stellar velocity dispersions predicted by cosmological NFW halos are less good representations, while the globular clusters allow a wide range of halo parameters. A suspected radial anisotropy of the stellar population aggravates the deviations. However, we find discrepancies with previous kinematical data, which we cannot resolve and may indicate a more complicated velocity pattern. Although one cannot conclusively demonstrate that the dark matter halo of NGC 3311 has a core rather than a cusp, a core seems to be preferred by the present data. A more complete velocity field and an analysis of the anisotropy is required to reach firm conclusions.Comment: 8 pages, 5 figures, abstract abridged, accepted for publication in A&

Splenic B1 B Cells Acquire a Proliferative and Anti-Inflamatory Profile During Pregnancy in Mice

B cells are a heterogeneous cell population with differential ontogeny, anatomical location, and functions. B1 B cells are a distinct subpopulation characterized by their unique capacity of self-renewal, the production of large quantities of IL-10, and the ability to secrete protective, anti-inflammatory natural antibodies (NAbs), presumably upon down-regulation of CD1d expression. Although natural antibodies are thought to be protective, due to their polyreactivity, their participation in certain autoimmune diseases has been suggested. In the context of pregnancy, the role of B1 B cells has been discussed controversially. While in human pregnancies B1 B cells and natural/polyreactive antibodies they produce are involved in the development of preeclampsia, in mice they promote healthy gestation and fetal protection. In this work, we aimed to functionally characterize the splenic B1 B cell population during pregnancy in mice. Functional enrichment analysis using only up-regulated transcripts from a transcriptomic profile performed on total splenic B cells from pregnant compared to non-pregnant mice showed augmented cell cycle and DNA replication pathways. Proliferation studies by flow cytometry showed augmented Ki-67 proliferation marker expression and percentages of B1 B cells. Furthermore, B1 B cells produced higher levels of IL-10 and lower levels of TNF-α leading to an increased IL-10/TNF-α ratio and showing an immunoregulatory phenotype. Finally, we observed lower expression of CD1d on B1 B cells, suggesting a higher capacity to produce NAbs in the context of pregnancy. In summary, our results showed not only an expanded and proliferative splenic B1 B cell population during pregnancy but also the acquisition of immunomodulatory capacities suggesting its critical role in the intricate process of pregnancy tolerance

Diversity and technological potential of lactic acid bacteria of wheat flours

Lactic acid bacteria (LAB) were analysed from wheat flours used in traditional bread making throughout Sicily (southern Italy). Plate counts, carried out in three different media commonly used to detect food and sourdough LAB, revealed a maximal LAB concentration of approximately 4.75 Log CFU g 1. Colonies representing various morphological appearances were isolated and differentiated based on phenotypic characteristics and genetic analysis by randomly amplified polymorphic DNA (RAPD)-PCR. Fifty unique strains were identified. Analysis by 16S rRNA gene sequencing grouped the strains into 11 LAB species, which belonged to six genera: Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Weissella. Weissella cibaria, Lactobacillus plantarum, Leuconostoc pseudomesenteroides and Leuconostoc citreum were the most prevalent species. The strains were not geographically related. Denaturing gradient gel electrophoresis (DGGE) analysis of total DNA of flour was used to provide a more complete understanding of the LAB population; it confirmed the presence of species identified with the culturedependent approach, but did not reveal the presence of any additional LAB species. Finally, the technological characteristics (acidifying capacity, antimicrobial production, proteolytic activity, organic acid, and volatile organic compound generation) of the 50 LAB strains were investigated. Eleven strains were selected for future in situ applications

Substructure and Scatter in the Mass-Temperature Relations of Simulated Clusters

Galaxy clusters exhibit regular scaling relations among their bulk properties. These relations establish vital links between halo mass and cluster observables. Precision cosmology studies that depend on these links benefit from a better understanding of scatter in the mass-observable scaling relations. Here we study the role of merger processes in introducing scatter into the $M$-$T_{\rm X}$ relation, using a sample of 121 galaxy clusters simulated with radiative cooling and supernova feedback, along with three statistics previously proposed to measure X-ray surface brightness substructure. These are the centroid variation ($w$), the axial ratio ($\eta$), and the power ratios ($P_{20}$ and $P_{30}$). We find that in this set of simulated clusters, each substructure measure is correlated with a cluster's departures $\delta \ln T_{\rm X}$ and $\delta \ln M$ from the mean $M$-$T_{\rm X}$ relation, both for emission-weighted temperatures $T_{\rm EW}$ and for spectroscopic-like temperatures $T_{\rm SL}$, in the sense that clusters with more substructure tend to be cooler at a given halo mass. In all cases, a three-parameter fit to the $M$-$T_{\rm X}$ relation that includes substructure information has less scatter than a two-parameter fit to the basic $M$-$T_{\rm X}$ relation.Comment: Accepted by ApJ, 10 pages, 10 figure

Redox regulation of cellular stress response in multiple sclerosis

Multiple sclerosis (MS) is an autoimmune-mediated neurodegenerative disease with characteristic foci of inflammatory demyelination in the brain, spinal cord, and optic nerves. Recent studies have demonstrated not only that axonal damage and neuronal loss are significant pathologic components of MS, but that this neuronal damage is thought to cause the permanent neurologic disability often seen in MS patients. Emerging finding suggests that altered redox homeostasis and increased oxidative stress, primarily implicated in the pathogenesis of MS, are a trigger for activation of a brain stress response. Relevant to maintenance of redox homeostasis, integrated mechanisms controlled by vitagenes operate in brain in preserving neuronal survival during stressful conditions. Vitagenes encode for heat shock proteins (Hsp) Hsp32, Hsp70, the thioredoxin and the sirtuin protein systems. In the present study we assess stress response mechanisms in the CSF, plasma and lymphocytes of control patients compared to MS patients. We found that the levels of vitagenes Hsp72, Hsc70, HO-1, as well as oxidative stress markers carbonyls and hydroxynonenals were significantly higher in the blood and CSF of MS patients than in control patients. In addition, an increased expression of Trx and sirtuin 1, together with a decrease in the expression of TrxR were observed. Our data strongly support a pivotal role for redox homeostasis disruption in the pathogenesis of MS and, consistently with the notion that new therapies that prevent neurodegeneration through nonimmunomodulatory mechanisms can have a tremendous potential to work synergistically with current MS therapies, unravel important targets for new cytoprotective strategies

A guanosine 5′-triphosphate-dependent protein kinase is localized in the outer envelope membrane of pea chloroplasts

A guanosine 5-triphosphate (GTP)-dependent protein kinase was detected in preparations of outer chloroplast envelope membranes of pea (Pisum sativum L.) chloroplasts. The protein-kinase activity was capable of phosphorylating several envelope-membrane proteins. The major phosphorylated products were 23- and 32.5-kilo-dalton proteins of the outer envelope membrane. Several other envelope proteins were labeled to a lesser extent. Following acid hydrolysis of the labeled proteins, most of the label was detected as phosphoserine with only minor amounts detected as phosphothreonine. Several criteria were used to distinguish the GTP-dependent protein kinase from an ATP-dependent kinase also present in the outer envelope membrane. The ATP-dependent kinase phosphorylated a very different set of envelope-membrane proteins. Heparin inhibited the GTP-dependent kinase but had little effect upon the ATP-dependent enzyme. The GTP-dependent enzyme accepted phosvitin as an external protein substrate whereas the ATP-dependent enzyme did not. The outer membrane of the chloroplast envelope also contained a phosphotransferase capable of transferring labeled phosphate from [-32P]GTP to ADP to yield (-32P]ATP. Consequently, addition of ADP to a GTP-dependent protein-kinase assay resulted in a switch in the pattern of labeled products from that seen with GTP to that typically seen with ATP

Chandra Cluster Cosmology Project II: Samples and X-ray Data Reduction

We discuss the measurements of the galaxy cluster mass functions at z=~0.05 and z=~0.5 using high-quality Chandra observations of samples derived from the ROSAT PSPC All-Sky and 400deg^2 surveys. We provide a full reference for the data analysis procedures, present updated calibration of relations between the total cluster mass and its X-ray indicators (T_X, Mgas, and Y_X) based on a subsample of low-z relaxed clusters, and present a first measurement of the evolving L_X-Mtot relation (with Mtot estimated from Y_X) obtained from a well-defined statistically complete cluster sample and with appropriate corrections for the Malmquist bias applied. Finally, we present the derived cluster mass functions, estimate the systematic uncertainties in this measurement, and discuss the calculation of the likelihood function. We confidently measure the evolution in the cluster comoving number density at a fixed mass threshold, e.g., by a factor of 5.0 +- 1.2 at M_500=2.5e14 h^-1 Msun between z=0 and 0.5. This evolution reflects the growth of density perturbations and can be used for the cosmological constraints complementing those from the distance-redshift relation.Comment: ApJ in press (Feb 10, 2009 issue); replacement to match accepted version, includes revisions in response to referee's and community comment

Social distancing in chronic migraine during the covid-19 outbreak: Results from a multicenter observational study

Background: The restrictions taken to control the rapid spread of COVID-19 resulted in a sudden, unprecedented change in people’s lifestyle, leading to negative consequences on general health. This study aimed to estimate the impact of such changes on migraine severity during 2020 March–May lockdown. Methods: Patients affected by migraine with or without aura, diagnosed by expert physicians, completed a detailed interview comprehensive of: assessment of migraine characteristics; measure of physical activity (PA) levels; measure of the intake frequency of main Italian foods; the Insomnia Severity Index (ISI) questionnaire investigating sleep disorders. Results: We included 261 patients with a mean age of 44.5 ± 12.3 years. During social distancing, 72 patients (28%) reported a headache worsening, 86 (33%) an improvement, and 103 (39%) a stable headache frequency. A significant decrease of the PA levels during COVID-19 quarantine in the whole study sample was observed (median total metabolic equivalent task (METs) decreased from 1170 to 510; p < 0.001). Additionally, a significant difference was reported on median ISI scores (from 7 to 8; p < 0.001), which were increased in patients who presented a stable or worsening headache. Conclusions: Our study confirmed that the restrictions taken during the pandemic have affected the practice of PA levels and sleep quality in migraine. Hence, PA and sleep quality should be assessed to find strategies for an improvement in quality of life

The Planetary Nebula Luminosity Function at the Dawn of Gaia

The [O III] 5007 Planetary Nebula Luminosity Function (PNLF) is an excellent extragalactic standard candle. In theory, the PNLF method should not work at all, since the luminosities of the brightest planetary nebulae (PNe) should be highly sensitive to the age of their host stellar population. Yet the method appears robust, as it consistently produces < 10% distances to galaxies of all Hubble types, from the earliest ellipticals to the latest-type spirals and irregulars. It is therefore uniquely suited for cross-checking the results of other techniques and finding small offsets between the Population I and Population II distance ladders. We review the calibration of the method and show that the zero points provided by Cepheids and the Tip of the Red Giant Branch are in excellent agreement. We then compare the results of the PNLF with those from Surface Brightness Fluctuation measurements, and show that, although both techniques agree in a relative sense, the latter method yields distances that are ~15% larger than those from the PNLF. We trace this discrepancy back to the calibration galaxies and argue that, due to a small systematic error associated with internal reddening, the true distance scale likely falls between the extremes of the two methods. We also demonstrate how PNLF measurements in the early-type galaxies that have hosted Type Ia supernovae can help calibrate the SN Ia maximum magnitude-rate of decline relation. Finally, we discuss how the results from space missions such as Kepler and Gaia can help our understanding of the PNLF phenomenon and improve our knowledge of the physics of local planetary nebulae.Comment: 12 pages, invited review at the conference "The Fundamental Cosmic Distance Scale: State of the Art and Gaia Perspective", to appear in Astrophysics and Space Scienc