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Hemopexin (Hpx) is a plasma glycoprotein which is expressed only in the liver. It is synthesized at a lower rate in the fetal liver than in the adult, and its level increases during acute infections. As shown here, a fragment of the human hemopexin promoter spanning from positions -130 to +22 relative to the cap site is sufficient to direct cell-specific transcription of a reporter gene. Within this segment a short sequence, located between positions -120 and -104, is responsible for this effect. This positive cis-acting element, the Hpx A site, interacts with a family of nuclear proteins, some of which are present only in hepatoma cells. The potential meaning of these complex DNA-protein interactions and the homology with elements present on the promoter of other liver-specific and acute phase genes are discussed.

Objective To assess the efficacy of recombinant human erythropoietin (rhEPO) in amyotrophic lateral sclerosis (ALS). Methods Patients with probable laboratory-supported, probable or definite ALS were enrolled by 25 Italian centres and randomly assigned (1:1) to receive intravenous rhEPO 40 000 IU or placebo fortnightly as add-on treatment to riluzole 100 mg daily for 12 months. The primary composite outcome was survival, tracheotomy or >23 h non-invasive ventilation (NIV). Secondary outcomes were ALSFRS-R, slow vital capacity (sVC) and quality of life (ALSAQ-40) decline. Tolerability was evaluated analysing adverse events (AEs) causing withdrawal. The randomisation sequence was computer-generated by blocks, stratified by centre, disease severity (ALSFRS-R cut-off score of 33) and onset (spinal or bulbar). The main outcome analysis was performed in all randomised patients and by intentionto-treat for the entire population and patients stratified by severity and onset. The study is registered, EudraCT 2009-016066-91. Results We randomly assigned 208 patients, of whom 5 (1 rhEPO and 4 placebo) withdrew consent and 3 ( placebo) became ineligible (retinal thrombosis, respiratory insufficiency, SOD1 mutation) before receiving treatment; 103 receiving rhEPO and 97 placebo were eligible for analysis. At 12 months, the annualised rate of death (rhEPO 0.11, 95% CI 0.06 to 0.20; placebo: 0.08, CI 0.04 to 0.17), tracheotomy or >23 h NIV (rhEPO 0.16, CI 0.10 to 0.27; placebo 0.18, CI 0.11 to 0.30) did not differ between groups, also after stratification by onset and ALSFRS-R at baseline. Withdrawal due to AE was 16.5% in rhEPO and 8.3% in placebo. No differences were found for secondary outcomes. Conclusions RhEPO 40 000 IU fortnightly did not change the course of ALS.

The phenomenon of acute phase (AP) response can be reproduced in vitro using cultured cells of hepatic origin by stimulation with the crude supernatant of activated monocytes (MoCM). Several monocyte-derived factors have been identified which might be responsible, alone or in combination, for the induction of AP response, but recently the attention has been focused on interleukin 6 (IL-6). We have previously shown that part of the AP response consists of the increase in the rate of transcription of the AP genes. Here we have treated the human hepatoma cell line Hep 3B with either crude MoCM or recombinant IL-6 and compared the effect of the two stimulants on the expression of both endogenous AP genes and recombinant plasmids introduced into the cells by transfection. The transfected plasmids contained the 5'-flanking region of AP genes fused to the coding region of the bacterial chloramphenicol acetyltransferase gene. We observe that the induction of mRNA accumulation of the endogenous genes corresponds to the transcriptional activation of the chloramphenicol acetyltransferase fusions. This is good evidence that the effect of IL-6 is totally or partially exerted at the level of transcription and that short segments of the 5'-flanking sequences of the inducible genes contain IL-6-responsive elements. Our results show that IL-6 is fully effective only on some of all the genes induced or repressed by MoCM, whereas others are only partially affected or totally nonresponsive.

Type II mixed cryoglobulinemia (CryoII) is an autoimmune disorder frequently associated to human hepatitis C virus (HCV) infection and characterized by the presence of cold-insoluble immunocomplexes containing IgM with rheumatoid activity. To identify disease-related epitopes, we screened a phage-displayed random peptide library using purified IgM from patients with HCV-associated CryoII(CryoII/HCV). A dominant population of phage isolates bearing the HPLAP pentapeptide consensus motif was identified and shown to be recognized by a nonrheumatoid IgM species strongly associated to CryoII/HCV. The phage-borne mimotopes (phagotopes) displayed a strong homology with an exposed extra-loop region of human lymphocyte activation 3 gene (LAG-3) product. Consistently, rabbit sera raised against a synthetic LAG-3 peptide efficiently recognized the selected phagotopes. Furthermore, one such phagotope was revealed to be a good immunogenic mimic of LAG-3 when injected into rabbits. IgM purified from CryoII/HCV patients' sera specifically reacted with the LAG-3 peptide in ELISA, and this binding was inhibited by the selected phagotopes. These results provide experimental support for a general strategy to identify novel autoantigens.

Background In multiple sclerosis (MS), determination of regional brain atrophy is clinically relevant. However, analysis of large datasets is rare because of the increased variability in multicenter data. Purpose To compare different methods to correct for center effects. To investigate regional gray matter (GM) volume in relapsing-remitting MS in a large multicenter dataset. Methods MRI scans of 466 MS patients and 279 healthy controls (HC) were retrieved from the Italian Neuroimaging Network Initiative repository. Voxel-based morphometry was performed. The center effect was accounted for with different methods: (a) no correction, (b) factor in the statistical model, (c) ComBat method and (d) subsampling procedure to match single-center distributions. By applying the best correction method, GM atrophy was assessed in MS patients vs HC and according to clinical disability, disease duration and T-2 lesion volume. Results were assessed voxel-wise using general linear model. Results The average residuals for the harmonization methods were 5.03 (a), 4.42 (b), 4.26 (c) and 2.98 (d). The comparison between MS patients and HC identified thalami and other deep GM nuclei, the cerebellum and several cortical regions. At single-center analysis, the thalami were always involved, whereas different other regions were found in each center. Cerebellar atrophy correlated with clinical disability, while deep GM nuclei atrophy correlated with T-2-lesion volume. Conclusion Harmonization based on subsampling more effectively decreased the residuals of the statistical model applied. In comparison with findings from single-center analysis, the multicenter results were more robust, highlighting the importance of data repositories from multiple centers.

A cross‐laboratory study on microbial fuel cells (MFC) which involved different institutions around the world is presented. The study aims to assess the development of autochthone microbial pools enriched from domestic wastewater, cultivated in identical single‐chamber MFCs, operated in the same way, thereby approaching the idea of developing common standards for MFCs. The MFCs are inoculated with domestic wastewater in different geographic locations. The acclimation stage and, consequently, the startup time are longer or shorter depending on the inoculum, but all MFCs reach similar maximum power outputs (55±22 μW cm−2) and COD removal efficiencies (87±9 %), despite the diversity of the bacterial communities. It is inferred that the MFC performance starts when the syntrophic interaction of fermentative and electrogenic bacteria stabilizes under anaerobic conditions at the anode. The generated power is mostly limited by electrolytic conductivity, electrode overpotentials, and an unbalanced external resistance. The enriched microbial consortia, although composed of different bacterial groups, share similar functions both on the anode and the cathode of the different MFCs, resulting in similar electrochemical output., The famous five: A cross‐laboratory study involving five institutions is presented. Each team runs a triplicate identical single chamber microbial fuel cell for 6 weeks varying only the inoculum. Electrochemical and operational parameters are collected. Microbiological analysis is carried out on the influent and effluent and on the electrodes. All the data collected are correlated by using robust statistics.