Assessment of Poly(ADP-ribose) Polymerase1 (PARP1) expression and activity in cells purified from blood and milk of dairy cattle

Poly(ADP-ribosyl)ation (PAR) is a post-translational protein modification catalysed by enzyme member of the poly(ADP-ribose) polymerases (PARPs) family. The activation of several PARPs is triggered by DNA strand breakage and the main PARP enzyme involved in this process is PARP1. Besides its involvement in DNA repair, PARP1 is involved in several cellular processes including transcription, epigenetics, chromatin re-modelling as well as in the maintenance of genomic stability. Moreover, several studies in human and animal models showed PARP1 activation in various inflammatory disorders. The aims of the study were (1) to characterize PARP1 expression in bovine peripheral blood mononuclear cells (PBMC) and (2) to evaluate PAR levels as a potential inflammatory marker in cells isolated from blood and milk samples following different types of infection, including mastitis. Our results show that (i) bovine PBMC express PARP1; (ii) lymphocytes exhibit higher expression of PARP1 than monocytes; (iii) PARP1 and PAR levels were higher in circulating PBMCs of infected cows; (iv) PAR levels were higher in cells isolated from milk with higher Somatic Cell Counts (SCC > 100,000 cells/mL) than in cells from milk with low SCCs. In conclusion, these findings suggest that PARP1 is activated during mastitis, which may prove to be a useful biomarker of mastitis

Non-invasive tool to assess heart rhythm in Zebrafish embryos

In the last years the zebrafish (Danio rerio) has emerged as model organism for cardiac research, in spite of the morphological differences with the human heart. In consequence of the similarity to humans in the early function, the zebrafish embryo has been suggested as an ideal model i) to study the molecular mechanism of cardiac development, and ii) to identify genes related to congenital cardiac defects in human [1]. The overall similarity of zebrafish embryos and human, in responses to human cardiotoxic drugs, was demonstrated, for example, in drug-induced cardiac arrhythmia [2]. For this reason, several methods have been developed to assess cardiac functions in zebrafish embryos [3,4]. Unfortunately, all these techniques suffer from drawbacks (time consuming, skillful operator are ended to perform the experiments) which limit their applications for large scale studies. The development in digital imaging has recently made analysis of cardiac functions in genetically modified transparent zebrafish embryos easier. This allowed to assess non-invasively heart rate variability in zebrafish embryos from videos of beating heart, but without measuring heartbeat rhythm, an important indicator of the cardiac function (heartbeat regularity is associated with cardiotoxicity in humans [1]), from power spectrum of heart signal. In the present study, we present a simple, non-invasive method that, by video-recording embryo images using confocal microscopy, and integrating image processing and power spectral analysis, allows to measure the heartbeat rhythm in zebrafish embryos heart chambers (atrium, ventricle, bulb) (Figure 1). The reliability of the herein proposed method was verified. Some embryos undergone treatment by tricaine, a cardiac anaesthetizing drug, in consequence of which a decrease of the heart rate is expected: the heartbeat regularity in tricaine- treated embryos determined from power spectral analysis decreased as compared to no-treated embryos. The results demonstrated that our method is able to assess the cardiac physiology, in term of heart rhythm, in zebrafish embryos

Decline in macrolide resistance rates among Streptococcus pyogenes causing pharyngitis in children isolated in Italy

Macrolides are often used to treat group A streptococcus (GAS) infections, but their resistance rates reached high proportions worldwide. The aim of the present study was to give an update on the characteristics and contemporary prevalence of macrolide-resistant pharyngeal GAS in Central Italy. A total of 592 isolates causing pharyngitis in children were collected in the period 2012-2013. Clonality was assessed by emm typing and pulsed-field gel electrophoresis (PFGE) for all macrolide-resistant strains and for selected susceptible isolates. Genetic determinants of resistance were screened by polymerase chain reaction (PCR). Forty-four GAS were erythromycin-resistant (7.4 %). Among them, 52.3 % and 50 % were clindamycin- and tetracycline-resistant, respectively. erm(B)-positive isolates (52.3 %) expressed the constitutive cMLSB phenotype. mef(A) and its associated M phenotype were recorded in 40.9 % of the cases. The remaining erm(A)-positive isolates expressed the iMLSB phenotype. Seventeen tetracycline-resistant isolates carried tet(M) and five isolates carried tet(O). Twenty-five emm types were found among all strains, with the predominance of emm types 12, 89, 1, and 4. Eleven emm types and 12 PFGE clusters characterized macrolide-resistant strains, with almost two-thirds belonging to emm12, emm4, and emm11. Macrolide-susceptible and -resistant emm types 12, 89, 11, and 4 shared related PFGE profiles. There was a dramatic decline in macrolide resistance in Central Italy among pharyngeal GAS isolates in 2012-2013 when compared to previous studies from the same region (p < 0.05), although macrolide consumption remained stable over the past 15 years. We observed a decrease in the proportion of macrolide-resistant strains within emm types commonly associated with macrolide resistance in the past, namely emm12, 1, and 89

Triphasic scaffolds for the regeneration of the bone-ligament interface

A triphasic scaffold (TPS) for the regeneration of the bone-ligament interface was fabricated combining a 3D fiber deposited polycaprolactone structure and a polylactic co-glycolic acid electrospun. The scaffold presented a gradient of physical and mechanical properties which elicited different biological responses from human mesenchymal stem cells. Biological test were performed on the whole TPS and on scaffolds comprised of each single part of the TPS, considered as the controls. The TPS showed an increase of the metabolic activity with culturing time that seemed to be an average of the controls at each time point. The importance of differentiation media for bone and ligament regeneration was further investigated. Metabolic activity analysis on the different areas of the TPS showed a similar trend after 7 days in both differentiation media. Total alkaline phosphatase (ALP) activity analysis showed a statistically higher activity of the TPS in mineralization medium compared to the controls. A different glycosaminoglycans amount between the TPS and its controls was detected, displaying a similar trend with respect to ALP activity. Results clearly indicated that the integration of electrospinning and additive manufacturing represents a promising approach for the fabrication of scaffolds for the regeneration of tissue interfaces, such as the bone-to-ligament one, because it allows mimicking the structural environment combining different biomaterials at different scales

The Blackholic energy and the canonical Gamma-Ray Burst IV: the "long", "genuine short" and "fake - disguised short" GRBs

(Shortened) [...] After recalling the basic features of the "fireshell model", we emphasize the following novel results: 1) the interpretation of the X-ray flares in GRB afterglows as due to the interaction of the optically thin fireshell with isolated clouds in the CircumBurst Medium (CBM); 2) an interpretation as "fake - disguised" short GRBs of the GRBs belonging to the class identified by Norris & Bonnell [...] consistent with an origin from the final coalescence of a binary system in the halo of their host galaxies with particularly low CBM density [...]; 3) the first attempt to study a genuine short GRB with the analysis of GRB 050509B, that reveals indeed still an open question; 4) the interpretation of the GRB-SN association in the case of GRB 060218 via the "induced gravitational collapse" process; 5) a first attempt to understand the nature of the "Amati relation", a phenomenological correlation between the isotropic-equivalent radiated energy of the prompt emission E_{iso} with the cosmological rest-frame \nu F_{\nu} spectrum peak energy E_{p,i}. In addition, recent progress on the thermalization of the electron-positron plasma close to their formation phase, as well as the structure of the electrodynamics of Kerr-Newman Black Holes are presented. An outlook for possible explanation of high-energy phenomena in GRBs to be expected from the AGILE and the Fermi satellites are discussed. As an example of high energy process, the work by Enrico Fermi dealing with ultrarelativistic collisions is examined. It is clear that all the GRB physics points to the existence of overcritical electrodynamical fields. In this sense we present some progresses on a unified approach to heavy nuclei and neutron stars cores, which leads to the existence of overcritical fields under the neutron star crust.Comment: 68 pages, 50 figures, in the Proceedings of the XIII Brazilian School on Cosmology and Gravitation, M. Novello, S.E. Perez-Bergliaffa, editor

SWIPE: a bolometric polarimeter for the Large-Scale Polarization Explorer

The balloon-borne LSPE mission is optimized to measure the linear polarization of the Cosmic Microwave Background at large angular scales. The Short Wavelength Instrument for the Polarization Explorer (SWIPE) is composed of 3 arrays of multi-mode bolometers cooled at 0.3K, with optical components and filters cryogenically cooled below 4K to reduce the background on the detectors. Polarimetry is achieved by means of large rotating half-wave plates and wire-grid polarizers in front of the arrays. The polarization modulator is the first component of the optical chain, reducing significantly the effect of instrumental polarization. In SWIPE we trade angular resolution for sensitivity. The diameter of the entrance pupil of the refractive telescope is 45 cm, while the field optics is optimized to collect tens of modes for each detector, thus boosting the absorbed power. This approach results in a FWHM resolution of 1.8, 1.5, 1.2 degrees at 95, 145, 245 GHz respectively. The expected performance of the three channels is limited by photon noise, resulting in a final sensitivity around 0.1-0.2 uK per beam, for a 13 days survey covering 25% of the sky.Comment: In press. Copyright 2012 Society of Photo-Optical Instrumentation Engineers. One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper are prohibite

The PU.1 transcription factor induces cyclin D2 expression in U937 cells [16]

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Clostridium botulinum spores and toxin in mascarpone cheese and other milk products

A total of 1,017 mascarpone cheese samples, collected at retail, were analyzed for Clostridium botulinum spores and toxin, aerobic mesophilic spore counts, as well as pH, a(w) (water activity), and Eh (oxidation-reduction potential). In addition 260 samples from other dairy products were also analyzed for spores and botulinum toxin. Experiments were carried out on naturally and artificially contaminated mascarpone to investigate the influence of different temperature conditions on toxin production by C. botulinum. Three hundred and thirty-one samples (32.5%) of mascarpone were positive for botulinal spores, and 7 (0.8%) of the 878 samples produced at the plant involved in an outbreak of foodborne botulism also contained toxin type A. The chemical-physical parameters (pH, a(w), Eh) of all samples were compatible with C. botulinum growth and toxinogenesis. Of the other milk products, 2.7% were positive for C. botulinum spores. Growth and toxin formation occurred in naturally and experimentally contaminated mascarpone samples after 3 and 4 days of incubation at 28 degrees C, respectively

Post-traumatic myocardial infarction with hemorrhage and microvascular damage in a child with myocardial bridge: is coronary anatomy actor or bystander?

We present the case of a 13 year old patient with myocardial bridge in left anterior descending coronary artery, who develops a myocardial infarction after a cardiothoracic trauma. About 24 hours after admission for trauma, an Electrocardiogram (ECG) showed an ST-segment elevation on anterior-lateral leads and QS complex referable to anterior-septal infarction, and an increase in troponin T serum levels was noted. An impaired left ventricular ejection fraction with diffuse regional wall motion abnormalities involving the left ventricular apex and interventricular septum were seen at transthoracic echocardiography. Contrast enhanced cardiac magnetic resonance showed a widespread myocardial edema and necrosis at the level of left ventricular apex and interventricular septum. Intramural hemorrhage and signs of microvascular damage were found mainly at the mid-ventricular level of the anteroseptal and anterior segments of myocardium. The coronary angiography revealed normal coronary arteries except for a myocardial bridge on distal part of left anterior descending coronary artery. A myocardial infarction with hemorrhage and microvascular damage was diagnosed, but the absence of a correspondence between site of the most severe myocardial injury and distal location of myocardial bridge was noted. Whether myocardial infarction and microvascular damage have been caused only by traumatic hit, or also by the contribution of myocardial bridge, is unknown. An intense constriction of left anterior descending coronary artery at the level of myocardial bridge could have determined thrombus formation with subsequent septal and distal embolization and myocardial infarction