Pivotal role of micro-CT technology in setting up an optimized lung fibrosis mouse model for drug screening

Idiopathic pulmonary fibrosis (IPF) is a progressive disease with no curative pharmacological treatment. The most used animal model of IPF for anti-fibrotic drug screening is bleomycin (BLM)-induced lung fibrosis. However, several issues have been reported: the balance among disease resolution, an appropriate time window for therapeutic intervention and animal welfare remain critical aspects yet to be fully elucidated. In this study, C57Bl/6 male mice were treated with BLM via oropharyngeal aspiration (OA) following either double or triple administration. The fibrosis progression was longitudinally assessed by micro-CT every 7 days for 4 weeks after BLM administration. Quantitative micro-CT measurements highlighted that triple BLM administration was the ideal dose regimen to provoke sustained lung fibrosis up to 28 days. These results were corroborated with lung histology and Bronchoalveolar Lavage Fluid cells. We have developed a mouse model with prolonged lung fibrosis enabling three weeks of a curative therapeutic window for the screening of putative anti-fibrotic drugs. Moreover, we have demonstrated the pivotal role of longitudinal micro-CT imaging in reducing the number of animals required per experiment in which each animal can be its own control. This approach permits a valuable decrease in costs and time to develop disease animal models

Modeling Neisseria meningitidis metabolism: from genome to metabolic fluxes

A genome-scale flux model for primary metabolism of Neisseria meningitidis was constructed; a minimal medium for growth of N. meningitidis was designed using the model and tested successfully in batch and chemostat cultures

Indocyanine-enhanced mouse model of bleomycin-induced lung fibrosis with hallmarks of progressive emphysema

The development of new drugs for idiopathic pulmonary fibrosis strongly relies on preclinical experimentation, which requires the continuous improvement of animal models and integration with in vivo imaging data. Here, we investigated the lung distribution of bleomycin (BLM) associated with the indocyanine green (ICG) dye by fluorescence imaging. A long-lasting lung retention (up to 21 days) was observed upon oropharyngeal aspiration (OA) of either ICG or BLM þ ICG, with significantly more severe pulmonary fibrosis, accompanied by the progressive appearance of emphysema-like features, uniquely associated with the latter combination. More severe and persistent lung fibrosis, together with a progressive air space enlargement uniquely associated with the BLM þ ICG group, was confirmed by longitudinal micro-computed tomography (CT) and histological analyses. Multiple inflammation and fibrosis biomarkers were found to be increased in the bronchoalveolar lavage fluid of BLM- and BLM þ ICG-treated animals, but with a clear trend toward a much stronger increase in the latter group. Similarly, in vitro assays performed on macrophage and epithelial cell lines revealed a significantly more marked cytotoxicity in the case of BLM þ ICG-treated mice. Also unique to this group was the synergistic upregulation of apoptotic markers both in lung sections and cell lines. Although the exact mechanism underlying the more intense lung fibrosis phenotype with emphysema-like features induced by BLM þ ICG remains to be elucidated, we believe that this combination treatment, whose overall effects more closely resemble the human disease, represents a valuable alternative model for studying fibrosis development and for the identification of new antifibrotic compounds.</p

ILC Vertex Tracker R&D

This document summarizes past achievements, current activities and future goals of the R&amp;D program aimed at the design, prototyping and characterization of a full detector module, equipped with monolithic pixel sensors, matching the requirements for the Vertex Tracker at the ILC. We provide a plan of activities to obtain a demonstrator multi-layered vertex tracker equipped with sensors matching the ILC requirements and realistic lightweight ladders in FY11, under the assumption that ILC detector proto-collaborations will be choosing technologies and designs for the Vertex Tracker by that time. The R&amp;D program discussed here started at LBNL in 2004, supported by a Laboratory Directed R&amp;D (LDRD) grant and by funding allocated from the core budget of the LBNL Physics Division and from the Department of Physics at UC Berkeley. Subsequently additional funding has been awarded under the NSF-DOE LCRD program and also personnel have become available through collaborative research with other groups. The aim of the R&amp;D program carried out by our collaboration is to provide a well-integrated, inclusive research effort starting from physics requirements for the ILC Vertex Tracker and addressing Si sensor design and characterization, engineered ladder design, module system issues, tracking and vertex performances and beam test validation. The broad scope of this program is made possible by important synergies with existing know-how and concurrent programs both at LBNL and at the other collaborating institutions. In particular, significant overlaps with LHC detector design, SLHC R&amp;D as well as prototyping for the STAR upgrade have been exploited to optimize the cost per deliverable of our program. This activity is carried out as a collaborative effort together with Accelerator and Fusion Research, the Engineering and the Nuclear Science Divisions at LBNL, INFN and the Department of Physics in Padova, Italy, INFN and the Department of Physics in Torino, Italy and the Department of Physics of Purdue University

Descriptive geometry in England: lost in translation

This chapter looks at the history of descriptive geometry in England, and why here it had such a short, and not a very fulfilling life. Having arrived to England in the immediate aftermath of the wars between England and France, its translation and attempts to introduce it into the educational system happened only after the 1840s. The lack of direct, implicit knowledge of the original technique, and some aspects of mistranslation, meant that the technique was never properly understood. Descriptive geometry is still mainly regarded as a drawing, rather than a mathematical technique in England, and has not been practised since the end of the nineteenth century. Polytechnic schools in England were another short-lived phenomena, and only of any significance and showing similarity with the French model in the second half of the twentieth century

A rapidly labelled RNA associated with DNA in HeLa cells

The nucleic acids of HeLa cells, labelled for 30 min with [3H]uridine are extracted by sodium dodecylsulphate-chloroform-isoamylalcohol and fractionated on methylated bovine serum albumin adsorbed on kieselguhr (MAK) columns and by Cs2SO4 density gradient centrifugation. A DNA-RNA complex is described which involves a fraction of labelled RNA resistant to ribonuclease at conditions which distinguish hybridized from free RNA. This complex is dissociated, like DNA-RNA hybrids, by heat treatment and does not appear to be an artificial association formed between DNA and non-specific RNA at the moment of cell lysis or during any of the subsequent biochemical steps. The heat dissociated RNA sediments in sucrose gradients with a peak at about 4-6 S, no radioactivity being detected in the heavy regions of the gradients. The limited size of these molecules is tentatively attributed to their breakage during extraction, or to their degradation due to the heat treatment used to free them from DNA. The true hybridized, ribonuclease-resistant structures appear to consist of short sequences of newly synthesized RNA, containing about 20-40 nucleotides. The RNA bound to the DNA at the time of extraction is of the order of 0.35 % of the total RNA synthesized during a 30-min incubation (this is probably a minimum value), and the proportion of DNA carrying such a fraction of RNA is about 0.03 %. © 1967.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

A Fraction of Newly Synthesized DNA of HeLa Cells

HeLa cells were labelled for 10 or 30 minutes with [3H]thymidine and the DNA fractionated with chloroform‐isoamylalcohol; almost 90% of the labelled DNA remained in the interphase whilst about 60% of total DNA (bulk) was extracted in the aqueous phase. When added to a column of methylated serum albumin adsorbed on kieselguhr the extracted DNA was eluted as a major labelled peak with 0.8 M NaCl but a small fraction could only be eluted at higher NaCl concentrations. This “tail” contained both bulk and recently labelled DNA. After a short pulse the specific activity of the DNA eluted at higher salt concentrations increased; this was confirmed when pooled fractions were concentrated on a caesium sulphate gradient. Electron microscopy and sucrose gradient centrifugation of fractions recovered on a Cs2SO4 gradient shows that the eluted DNA was polydisperse (peak at about 18 S) with molecular dimensions of a few to almost 20 μ. When [3H]uridine was used as a label, both labelled DNA and rapidly labelled RNA could be studied simultaneously in the same way: these experiments indicated that in the fractions analysed, the radioactivity due to RNA and to DNA was not carried on the same molecules, suggesting the mutual exclusion of replication and transcription. Copyright © 1967, Wiley Blackwell. All rights reservedSCOPUS: ar.jinfo:eu-repo/semantics/publishe

Focus on the role of substance P in chronic urticaria

Abstract Background Emerging data have strengthened the importance of substance P (SP) as a proinflammatory mediator in human pathology. A role for SP in the pathogenesis of urticaria has long been hypothesized. Methods Literature data regarding the possible role of SP in chronic urticaria/chronic spontaneous urticaria (CSU) have been reviewed and summarized in this manuscript. This review is based on pertinent articles that were retrieved by a selective literature search in the PubMed database. Articles in English published up to July 2018 were taken into consideration. Results Recent studies in patients with CSU have demonstrated that circulating levels of SP are significantly elevated, in correlation with disease severity, and that SP-positive basophils are upregulated. SP has been shown to trigger degranulation in basophils derived from CSU patients. Moreover, SP can be involved in pseudoallergic reactions and may act as a histamine-releasing factor in a subset of patients with CSU. Current evidence suggests that the biological activity of SP can be exerted not only through the conventional NK-1 receptor but also through the recently identified Mas-related G protein-coupled receptors. MRGPRX2 can cause mast cell activation and has been found to be upregulated in the skin of patients with severe chronic urticaria. Conclusions Many findings seem to support the pathogenic involvement of SP in chronic urticaria/CSU. However, further studies are necessary to elucidate the role of SP as a mediator in CSU pathogenesis and a potential new therapeutic target