Analysis of the coupled effect of steel studs and surface emissivity on internal insulation systems performance

Many kinds of insulation systems have been developed and applied over the years to all the constructive elements of the building, but the two most used strategies remain the external and internal insulation of vertical walls. However, about the latter often a significant issue is neglected: the overestimation of the thermal performance by disregarding the contribution of construction elements. Usually a uniform stratigraphy of the wall is considered and the evaluation of the performance of a non-uniform one leads to erroneous results about the overall behavior of the system. In this paper, we developed a different approach considering the presence of the steel studs used to attach this package to the existing wall and their influence on the thermal behavior of the structure. Through both experimental and numerical analysis, the possible application of low-e sheets inside the air cavity in various configurations and with different thicknesses of insulation is also taken into account. Results showed that neglecting the presence of the steel studs leads to an erroneous evaluation of the conductance of the refurbished wall with errors reaching up to 28.0% in low-e high-insulated cases. This work highlights how careful the designers have to be when using standard formulas to compute the thermal resistance of internal insulation wall systems

genetic optimization for economic feasibility of refurbishment in buildings

Abstract This paper investigates the possibility to enhance the thermal characteristics of a building fabric taking into account the economic feasibility in order to avoid useless expensive interventions. The problem consists in realizing an internal insulation system for a single apartment located in Trieste. A genetic optimization has been implemented in order to search a large number of solutions with multiple objects and constraints. EnergyPlus has been used for dynamic building and plant simulation and modeFRONTIER for optimization loop setup. The objectives are the used heating energy consumption and the life cycle refurbishment cost, using a net present value approach. An additional objective has been added in order to guarantee the maximization of the interior useful floor area, which is directly influenced by the internal insulation thickness. Different approaches have been compared, considering actions dealing with opaque surfaces only or considering the replacement of existing windows. The paper demonstrates that an automatic optimization approach can help designers to identify the best suited intervention for building refurbishment

-Tetrachlorodibenzo-p-dioxin-Mediated Impairment of B Cell Differentiation Involves Dysregulation of Paired Box 5 (Pax5) Isoform, Pax5a

ABSTRACT The persistent environmental contaminant and immunotoxicant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), markedly suppresses humoral immune responses. We recently reported impaired down-regulation of paired box 5 (Pax5), a repressor of B cell differentiation and concomitant suppression of the IgM response by TCDD in the murine CH12.LX B cell line. The objectives of the current study were to determine the impact of TCDD treatment on molecular outcomes characteristic of terminal B cell differentiation and to assess the role that Pax5 isoforms plays in the suppression of B cell differentiation by TCDD. In this study, we show that the highly abundant fulllength Pax5 isoform, Pax5a, and at least two additional modestly expressed Pax5 isoforms were expressed in CH12.LX and splenic B cells. In lipopolysaccharide (LPS)-activated B cells, all of the identified Pax5 isoforms were synchronously down-regulated, and in the presence of TCDD cotreatment they were abnormally and synchronously elevated, suggesting a common mechanism of regulation. Furthermore, B cell differentiation markers X-box protein-1 and major histocompatibility complex class II showed that the levels to which Pax5 was derepressed by TCDD were sufficient to impair B cell differentiation and immunoglobulin gene expression. Confirming the involvement of Pax5, ectopic expression of Pax5a in the LPS-activated CH12.LX cells closely mimicked the suppression of the IgM response by TCDD. In summary, our results demonstrate that Pax5a has a critical role in both the TCDD-mediated impairment of B cell differentiation and the suppression of the humoral immune response. Suppression of primary humoral immune responses is one of the most sensitive sequela associated with exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a ubiquitous environmental contaminant. This suppression is characterized by a striking reduction in plasma cell formation and IgM secretion, and it is mediated through a direct effect by TCDD on B cells Activation by antigen or via polyclonal activators (e.g., 463 lipopolysaccharide; LPS) induces B cells to undergo cycles of intense proliferation (i.e., clonal expansion) followed by terminal differentiation into plasma cells. Terminally differentiated plasma cells specialize in secretion of antigen-specific Ig. A number of phenotypic changes distinguish terminally differentiated plasma cells from the resting B cells Materials and Methods Chemicals. TCDD, in 100% dimethyl sulfoxide (DMSO), was purchased from AccuStandard (New Heaven, CT). DMSO and LPS were purchased from Sigma-Aldrich (St. Louis, MO). Mice. Virus-free, female B6C3F1mice (six weeks old) were purchased from Charles River (Portage, MI). On arrival, mice were randomly grouped with five per plastic cage on sawdust bedding. Mice had free access to food (Purina Certified Laboratory Chow) and water at all times. The mouse holding rooms were maintained at 21 to 24°C with 40 to 60% relative humidity on a 12-h light/dark cycle. All of the experimental procedures and conditions were performed according to the guidelines of the All University Committee on Animal Use and Care at Michigan State University (East Lansing, MI). Cell Line. The CH12.LX B cell line was derived from the murine B cell lymphoma, CH12, which arose in B10.H-2aH-4bP/Wts mice (B10.A ϫ B10.129) and has been characterized previously Flow Cytometric Analysis. Cells were harvested from culture at the indicated times by centrifugation at 300g for 10 min at 4°C, washed twice in ice-cold 1ϫ Hanks' balanced-salt solution, and stained using the BD Cytofix/Cytoperm kit (BD Pharmingen, San Diego, CA) according to the manufacturer's instructions. In brief, cells were incubated with 1 g/10 6 cells of purified rat anti-mouse CD16/CD32 monoclonal antibody (BD Pharmingen) for 15 min at 4°C to prevent nonspecific binding and then stained with surface marker detection antibody [anti-fluorescein isothiocyanate (FITC)-conjugated mouse anti-mouse I-A P or anti-allophycocyanin-conjugated anti-mouse CD19] or a respective isotype control (BD Pharmingen). To exclude nonviable cells, 2 l of 7-aminoactinomycin D (7-AAD) solution (Sigma-Aldrich) containing 1 mg of 7-AAD, 50 l methanol, and 950 l of Hanks' balanced-salt solution were added simultaneously with detection antibodies to the cells in 50 l of staining buffer. The cells were then fixed, washed, and maintained in staining buffer containing 10 mM actinomycin D (Sigma-Aldrich) to prevent 7-AAD leakage from fixed cells. For the detection of nuclear Pax5 protein, cells were fixed and permeabilized before staining with anti-Pax5 antibody or isotype control (Santa Cruz Biotechnology, Inc., Santa Cruz, CA). Fluorescence detection was performed using a BD FACSCalibur flow cytometer (BD Biosciences, San Jose, CA) on 10,000 viable cells per sample. Data analysis was performed using BD CellQuest Pro software (BD Biosciences). In Vitro Polyclonal IgM Antibody-Forming Cell Response. Single-cell suspensions of splenocytes from naive mice were adjusted to 1 ϫ 10 6 cells/ml in RPMI 1640 medium (Invitrogen) supplemented with 10% bovine calf serum (HyClone Laboratories), 100 units/ml penicillin, 100 g/ml streptomycin, and 50 M 2-mercaptoethanol. Spleen cells were transferred to 48-well culture plates in 500-l aliquots with four wells per treatment group. TCDD (3, 10, or 30 nM) and/or vehicle (0.01% DMSO) were added directly to each well in 5-l aliquots. The splenocytes were sensitized with 10 g/ml LPS and cultured for 3 days in a pressurized chamber at 5.0 psi containing 10% O 2 , 7% CO 2 , and 83% N 2 gas mixture with continuous rocking at 37°C. Enumeration of the IgM antibody-forming cell (AFC) response was performed using trinitrophenol-haptenated sheep red blood cells as described previously the cell mixture was poured onto a 100 ϫ 15-mm Petri dish and immediately covered with a 45 ϫ 50-mm glass coverslip. Upon solidification of the agar, the Petri dishes were placed in a humidified 37°C incubator overnight to allow for plaque formation. The number of splenocytes from each spleen was determined using a Coulter Counter (Beckman Coulter, Fullerton, CA). Results are expressed as AFC/10 6 viable splenocytes Ϯ S.E. Splenocyte viability was measured using pronase (EMD Biosciences, San Diego, CA) as described previously Purification of Splenic B Cells. Spleens were removed aseptically and made into a single-cell suspension. B cells were then isolated using the B Cell Isolation Kit (Miltenyi Biotec Inc., Auburn, CA) according to the manufacturer's protocol. In brief, 40 l of MACS buffer (phosphate-buffered saline solution, pH 7.2, supplemented with 0.5% bovine serum albumin and 2 mM EDTA, 4°C) per 10 7 cells was used to suspend the splenocytes, and 10 l of Biotin-Antibody Cocktail (Miltenyi Biotec Inc.) per 10 7 cells was added to label non-B cells. After incubation at 4 to 8°C for 10 min, 30 l of buffer and 20 l of Anti-Biotin Microbeads (Miltenyi Biotec Inc.) per 10 7 cells were added. The cell suspension was incubated for another 15 min at 4 to 8°C, washed with 10 to 20 times the labeling volume, and then centrifuged at 300g for 5 min. The cell pellet was finally resuspended in 500 l of buffer per 1 ϫ 10 8 cells, and passed through the prerinsed LS column (Miltenyi Biotec Inc.), followed by four washes of the column with 3 ml of buffer. The entire effluent containing the purified B cell fraction was collected. The purity of the isolated B cells was evaluated using flow cytometry to enumerate the percentage of CD19 ϩ cells through directly labeling with FITC-conjugated anti-CD19 antibody (BD Biosciences). Real-Time Polymerase Chain Reaction. Total RNA was isolated from naive or LPS-activated cells using a SV Total RNA Isolation kit (Promega, Madison, WI). To synthesize cDNA, 1000 ng/ sample of total RNA was incubated with 600 ng of random primer (Invitrogen) in 10 l of endonuclease-free water at 70°C for 10 min, cooled on ice for 10 min, and reverse transcribed in 20 l of 1ϫ First-Strand Synthesis buffer (Invitrogen), containing 0.2 mM deoxynucleotide triphosphates, 10 mM dithiothreitol, and 200 units of SuperScript II reverse transcriptase (Invitrogen). The reaction mixture was incubated at 42°C for 60 min, and the reaction was stopped by incubation at 75°C for 15 min. Real-time polymerase chain reaction (PCR) detection was performed using TaqMan primers and probes Isolation of Pax5a cDNA. cDNA was obtained by PCR amplification of total RNA isolated from CH12.LX cells. Total RNA was extracted with the SV Total RNA Isolation kit (Promega), and firststrand cDNA was synthesized by reverse transcription (RT) using a High Capacity cDNA Reverse Transcription kit (Applied Biosystems). PCR was carried out as follows: one initial 2-min denaturing step at 94°C followed by denaturation for 30 s at 94°C, annealing for 30 s at 58°C, and extension for 90 s at 72°C during 30 cycles in the presence of Pfu polymerase (Stratagene, La Jolla, CA) under the conditions suggested by the supplier (at a final primer concentration of 0.5 M). Sequences of the primers designed for this PCR reaction were as follows: 5Ј-CTGTCCATTTCATCAAGTCCTGA-3Ј and 5Ј-ACC-GTCACTACCCTCAGAG-3Ј. The resulting PCR product of 1.2 kilobase was separated in 1% agarose gel electrophoresis in 1ϫ TBE buffer (89 mM Tris, 89 mM boric acid, and 2 mM EDTA, pH 8.3) and stained with ethidium bromide at 1 g/ml. The PCR fragment was eluted from agarose using the QIAquick gel extraction kit (QIAGEN, Valencia, CA). Cloning and Ectopic Expression for Pax5a cDNA. Pax5a cDNA, generated as described above, was inserted into the pcDNA 3.1 V5-His-TOPO vector (Invitrogen) according to the manufacturer's protocol. The new Pax5a-V5-His tag vector sequence was confirmed by sequencing. Additional PCR reactions, in which Pax5a-V5-His vector was used as template, were performed to generate a new Pax5a expression plasmid, Pax5a-V5-His-GFP. The primer used for 5Ј-end cloning was as follows: 5Ј-CTC ACT ATA GGG AGA TCT AAG CTG GCT AGT-3Ј (BglII restriction site is underlined); and the primer used for the 3Јend cloning was as follows: 5Ј-TGA TCA GCG GGT TTA AAA GCT TTG GGA TGG TG-3Ј (HindIII restriction site is underlined). PCR products of 1.38 kilobase were obtained and isolated as described above. The Pax-5a-V5-His PCR fragment was then cloned into the vector phCMV-C-GFP (Genlantis, San Diego, CA) by a standard ligation reaction using T4-DNA ligase (New England Biolabs, Ipswich, MA). The phCMV vector from Genlantis contains an optimized cytomegalovirus promoter and intron sequences and a kanamycin/neomycin resistance gene for generation of stable cell lines. Transfection of Pax5a-V5-His-GFP. CH12.LX cells (2.5 ϫ 10 6 ) were transfected with 5 g of each plasmid using Amaxa Nucleofector (Amaxa Inc., Gaithersburg, MD). Cells, DNA, and 100 l of 90:20 (solution V supplement) were mixed and electroporated using program A-20 following the manufacturer's recommendations. A total of TCDD Impairment of B Cell Differentiation by Altered Pax5 465 2.5 ϫ 10 7 cells was used in each experiment. The backbone plasmid, phCMV-C-GFP, was used as a control in all of the transfection experiments. After electroporation, cells were incubated at 37°C in an atmosphere of 5% CO 2 for 8 h. Eight hours after transfection, cells were collected and sorted by flow cytometry for green fluorescent protein (GFP)-expressing cells. Cells that showed fluorescence above the level of fluorescence of naive CH12.LX cells were isolated using a BD FACSCalibur. The isolated cells were counted and LPS-activated for assessments of IgM secretion as determined by enzymelinked immunosorbent assay (ELISA). An aliquot of the sorted cells was also used to prepare protein extracts to determine endogenous/ ectopic Pax5a levels by Western blotting and by flow cytometry. IgM ELISA. IgM ELISA was performed as described previously Western Blotting. Proteins were extracted from transfected cells before and after sorting by flow cytometry. Protein extracts were resolved on 4 to 12% Nu-Page gradient gel (Invitrogen) and then transferred to a nitrocellulose membrane (GE Healthcare, Piscataway, NJ). Immunoblotting was performed using anti-␤-actin and anti-Pax5 antibodies (Santa Cruz Biotechnology, Inc.). SuperSignal West Pico reagent (Pierce, Rockford, IL) was used for protein detection. Statistical Analysis of Data. Mean Ϯ S.E. was determined for each treatment group of a given PCR or ELISA experiment. Statistical differences between groups in each experiment were determined by a two-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test for PCR experiments and by a one-way ANOVA followed by Bonferroni's post hoc test for ELISA experiments. Results TCDD Decreases the Levels of XBP-1 in LPS-Activated B Cells. Previous studies in LPS-activated CH12.LX cells demonstrated robust suppression of the humoral IgM response by TCDD, which correlated with a marked reduction in XBP-1 protein levels Additional studies were performed to determine whether LPS and/or TCDD treatment affects the post-transcriptional modification of XBP-1. Transcriptional activity of XBP-1 is known to be enhanced by a splicing event, which removes a 26-nucleotide-long fragment from XBP-1 mRNA, resulting in an open reading frame shift and yielding a larger XBP-1 466 Schneider et al. protein that is more potent as a transcription factor TCDD Attenuates Cell Surface MHC Class II DownRegulation in LPS-Activated CH12.LX Cells. An additional event closely associated with terminal differentiation of B cells is the down-regulation of MHC class II on the cell membrane. Levels of surface MHC class II in LPS-activated CH12.LX cells and in splenic B cells (i.e., CD19 ϩ ), in the presence and absence of TCDD, were monitored by flow cytometry. LPS activation induced a down-regulation of MHC class II on the CH12.LX cell surface between 24 and 72 h of culture compared with the time-matched naive control TCDD Attenuates the LPS-Induced Down-Regulation of Pax5 Protein in LPS-Activated CH12.LX Cells. We also examined the impact of TCDD on Pax5, a repressor of B cell differentiation. Expression of Pax5 protein was characterized in splenic B cells and CH12.LX cells by flow cytometry facilitating the evaluation of individual cells in contrast to prior Western blot studies that we performed exclusively in CH12.LX cells Characterization of Pax5 Isoforms in CH12.LX Cells. In light of the altered Pax5 expression observed by flow cytometry and by real-time PCR, studies were undertaken to further characterize the effect of TCDD on Pax5 regulation. In particular, we examined the role of alternative splicing in the deregulation of Pax5 by TCDD Additional studies performed in CH12.LX cells showed that the levels of amplicons I, II, and III were down-regulated by LPS between 24 and 72 h, as determined by real-time quantitative PC

Identifying frequency decorrelated dust residuals in B-mode maps by exploiting the spectral capability of bolometric interferometry

Astrophysical polarized foregrounds represent the most critical challenge in Cosmic Microwave Background (CMB) B-mode experiments. Multi-frequency observations can be used to constrain astrophysical foregrounds to isolate the CMB contribution. However, recent observations indicate that foreground emission may be more complex than anticipated. We investigate how the increased spectral resolution provided by band splitting in Bolometric Interferometry (BI) through a technique called spectral imaging can help control the foreground contamination in the case of unaccounted Galactic dust frequency decorrelation along the line-of-sight. We focus on the next generation ground-based CMB experiment CMB-S4, and compare its anticipated sensitivities, frequency and sky coverage with a hypothetical version of the same experiment based on BI. We perform a Monte-Carlo analysis based on parametric component separation methods (FGBuster and Commander) and compute the likelihood on the recovered tensor-to-scalar ratio. The main result of this analysis is that spectral imaging allows us to detect systematic uncertainties on r from frequency decorrelation when this effect is not accounted for in component separation. Conversely, an imager would detect a biased value of r and would be unable to spot the presence of a systematic effect. We find a similar result in the reconstruction of the dust spectral index, where we show that with BI we can measure more precisely the dust spectral index also when frequency decorrelation is present. The in-band frequency resolution provided by BI allows us to identify dust LOS frequency decorrelation residuals where an imager of similar performance would fail. This opens the prospect to exploit this potential in the context of future CMB polarization experiments that will be challenged by complex foregrounds in their quest for B-modes detection.Comment: 13 Pages, 15 figures, 4 tables. Submitted to A&

A chemically etched corrugated feedhorn array for D-band CMB observations

We present the design, manufacturing, and testing of a 37-element array of corrugated feedhorns for Cosmic Microwave Background CMB) measurements between 140 and 170 GHz. The array was designed to be coupled to Kinetic Inductance Detector arrays, either directly (for total power measurements) or through an orthomode transducer (for polarization measurements). We manufactured the array in platelets by chemically etching aluminum plates of 0.3 mm and 0.4 mm thickness. The process is fast, low-cost, scalable, and yields high-performance antennas compared to other techniques in the same frequency range. Room temperature electromagnetic measurements show excellent repeatability with an average cross polarization level about − 20 dB, return loss about − 25 dB, first sidelobes below − 25 dB and far sidelobes below − 35 dB. Our results qualify this process as a valid candidate for state-of-the-art CMB experiments, where large detector arrays with high sensitivity and polarization purity are of paramount importance in the quest for the discovery of CMB polarization B-modes

Measuring CMB spectral distortions from Antarctica with COSMO: blackbody calibrator design and performance forecast

COSMO is a ground-based instrument to measure the spectral distortions (SD) of the Cosmic Microwave Background (CMB). In this paper, we present preliminary results of electromagnetic simulations of its reference blackbody calibrator. HFSS simulations provide a calibrator reflection coefficient of R∼ 10 - 6, corresponding to an emissivity ϵ= 1 - R= 0.999999. We also provide a forecast for the instrument performance by using an ILC-based simulation. We show that COSMO can extract the isotropic Comptonization parameter (modeled as | y| = 1.77 · 10 - 6) as | y| = (1.79 ± 0.19) · 10 - 6, in the presence of the main Galactic foreground (thermal dust) and of CMB anisotropies, and assuming perfect atmospheric emission removal

An early European experience with transapical off-pump mitral valve repair with NeoChord implantation

OBJECTIVES: Transapical off-pump NeoChord repair is a novel minimally invasive surgical procedure to treat degenerative mitral valve regurgitation. The aim was to evaluate 1-year clinical results of the NeoChord procedure in a consecutive cohort of patients. METHODS: Between February 2013 and July 2016, 213 patients were enrolled in the NeoChord Independent International Registry. All patients presented severe mitral regurgitation due to flail/prolapse of 1 or both leaflets, and they all completed postoperative echocardiographic assessment up to 1 year. We identified the primary end point as composed of procedural success, freedom from mortality, stroke, reintervention, recurrence of severe mitral regurgitation, rehospitalization and decrease of at least 1 New York Heart Association functional class at 1-year follow-up. We also compared outcomes according to the anatomical classification (Type A: isolated central posterior leaflet disease; Type B: posterior multisegment disease; Type C: anterior, bileaflet, paracommissural disease with/without leaflet/annular calcifications). RESULTS: The median age was 68 years (interquartile range 56-77), and the median EuroSCORE II was 1.05% (interquartile range 0.67-1.76). The number of Type A, B and C patients was 82 (38.5%), 98 (46%) and 33 (15.5%), respectively. Procedural success was achieved in 206 (96.7%) patients. At 1-year follow-up, overall survival was 98 ± 1%. Composite end point was achieved in 84 ± 2.5% for the overall population and 94 ± 2.6%, 82.6 ± 3.8% and 63.6 ± 8.4% in Type A, Type B and Type C patients, respectively (P < 0.0001). CONCLUSIONS: These results demonstrate that the NeoChord procedure is safe, effective and reproducible. Clinical and echocardiographic efficacy is maintained up to 1 year with significant differences among the anatomical groups. Specific anatomical selection criteria are necessary to achieve stable results

CAS: centre for advanced studies

The JRC‘s Centre for Advanced Studies (CAS) was created in 2016 to help improve and bridge the interface between science and policy in order to enhance the JRC‘s capacity to better inform and influence the regulatory frameworks needed to address the new and emerging societal challenges confronting the EU and our societies as a whole. By creating the conditions necessary for innovative and interdisciplinary research, as well as offering a creative and generative space in which ideas and knowledge in emerging thematic fields across different scientific and technological disciplines can thrive and flourish, CAS has become an incubator for formal inquiry, stimulating ideas and activities and providing the JRC with new insights, data projections and solutions for the increasingly complex medium and longterm challenges facing the EU, especially in the fields of demography, big data and digital transformation. Through the performance of advanced, cutting edge research, ranging from applied research to topics of a more academic character, all within a stimulating trans- and interdisciplinary environment, CAS allows external researchers and scientists to work together with the JRC to explore and exchange new ideas and knowledge on scientific research in emerging fields of strategic societal importance, which might otherwise fall outside the policy support activities undertaken by the JRC on behalf of the European Commission

The COSmic Monopole Observer (COSMO)

The COSmic Monopole Observer (COSMO) is an experiment to measure low-level spectral distortions in the isotropic component of the Cosmic Microwave Background (CMB). Deviations from a pure blackbody spectrum are expected at low level (< 1 ppm) due to several astrophysical and cosmological phenomena, and promise to provide important independent information on the early and late phases of the universe. They have not been detected yet, due to the extreme accuracy required, the best upper limits being still those from the COBE-FIRAS mission. COSMO is based on a cryogenic differential Fourier Transform Spectrometer, measuring the spectral brightness difference between the sky and an accurate cryogenic blackbody. The first implementation of COSMO, funded by the Italian PRIN and PNRA programs, will operate from the Concordia station at Dome-C, in Antarctica, and will take advantage of a fast sky-dip technique to get rid of atmospheric emission and its fluctuations, separating them from the monopole component of the sky brightness. Here we describe the instrument design, its capabilities, the current status. We also discuss its subsequent implementation in a balloon-flight, which has been studied within the COSMOS program of the Italian Space Agency