Development of an N-1 perfusion process and optimized scale-down models for implementation in a platform CHO cell culture manufacturing process

The use of N-1 perfusion, coupled with high-inoculum fed batch in CHO cell culture manufacturing processes, has been shown to increase volumetric productivity and shorten the duration of the fed-batch production phase. Implementation of N-1 perfusion as part of a platform process requires the ability to screen multiple clones and to optimize media and process parameters in a high-throughput manner. We have developed an N-1 perfusion process, along with a series of scale-down models for N-1 perfusion using shake flasks, cell culture tubes, and deep-well plates. Process parameters for scale-down models were optimized to maximize comparability of growth profiles and cell culture performance relative to 5L N-1 perfusion bioreactors. Scale-down models were used to inoculate fed-batch experiments in Ambr15 micro-bioreactors at high seeding density, in order to compare growth and productivity profiles to those observed in 5L bench scale bioreactors. Multiple cell lines derived from different CHO hosts were evaluated in order to verify the robustness of the scale-down models. Results demonstrated that cell growth and viability in the optimized scale-down models were comparable to those observed in 5L N-1 perfusion bioreactors. Furthermore, growth, productivity, and product quality profiles from high-inoculum fed-batch experiments were comparable regardless of inoculum source. Optimized scale down models of N-1 perfusion, coupled with Ambr15 fed-batch production micro-bioreactors, have now been integrated into a high-throughput and robust workflow to enable DOE and screening experiments for clone selection, media development and parameter optimization in a platform N-1 perfusion process for monoclonal antibody manufacturing

Differences in the robustness of clusters involving the Mycobacterium tuberculosis strains most frequently isolated from immigrant cases in Madrid

AbstractTuberculosis cases infected by the same Mycobacterium tuberculosis (MTB) strain are considered to be clustered and involved in a transmission chain. Large clusters are assumed to represent active transmission chains in a population. In the present study, we focused on the analysis of large clusters defined by IS6110-restriction fragment length polymorphism (RFLP) typing in the immigrant population in Madrid. We identified 12 large clusters (involving 43% of the isolates) comprising 4–23 representatives. We proposed a gradient of epidemiological certainty for these large clusters. For a cluster to be considered robust and a good indicator of recent transmission, the MTB strain involved should not have been identified in a geographically and epidemiologically unrelated population and the cluster had to be re-confirmed by another highly discriminative molecular marker (MIRU-VNTR). The clusters that we discovered were classified into three categories: high, intermediate and low expected epidemiological value. In the largest cluster in the study (cluster M6; 23 representatives), failures by both criteria were identified: the representative seven-band RFLP pattern was also the most prevalent in the unrelated population (25 cases) and the cluster was fully split by MIRU-15, suggesting a lack of epidemiological value. The RFLP pattern representative of this cluster was also identified in 64 isolates from five countries in the Latin American genotype database, and again proved to be heterogeneous according to the MIRU-15 analysis. Specific analysis of large clusters, combined with the application of criteria for evaluating their robustness, could help identify uninformative clusters and target epidemiological resources towards those clusters with higher expected epidemiological value

Three-Dimensional Printing and Navigation in Bone Tumor Resection

One of the most promising advances raised by the current computer age is performing research “in silico,” which means computer-assisted. The objective of this chapter is firstly to evaluate if a 3D in-silico model of an oncological patient could be used to make a 3D-printed prototype in real scale, discriminating precisely healthy tissues, tumoral tissues and oncological margins. Secondly, the objective is to evaluate if this prototype could be representative enough to allow testing osteotomies under navigated guidance based on images. A tumor resection for a patient with diagnosed metaphyseal osteosarcoma of the proximal tibia was transferred into a rapid prototyping model, fabricated using 3D printing and representing different structures in different colors. The planned osteotomy was executed using Stryker Navigator to guide the cutting saw and the prototype was opened to verify the precision of the performed osteotomy. Both osteotomy planes showed successful correspondence with the safe margin, with a maximum error of 1 mm. The application of these techniques in general orthopedics would help to reduce the incidence of unforeseen intraoperative failures, contributing to obtain predictable surgical procedures. This would implement a new way of performing development, research and training in orthopedics and traumatology by in-silico technology

Non-parametric deprojection of NIKA SZ observations: Pressure distribution in the Planck-discovered cluster PSZ1 G045.85+57.71

The determination of the thermodynamic properties of clusters of galaxies at intermediate and high redshift can bring new insights into the formation of large-scale structures. It is essential for a robust calibration of the mass-observable scaling relations and their scatter, which are key ingredients for precise cosmology using cluster statistics. Here we illustrate an application of high resolution $(< 20$ arcsec) thermal Sunyaev-Zel'dovich (tSZ) observations by probing the intracluster medium (ICM) of the \planck-discovered galaxy cluster \psz\ at redshift $z = 0.61$, using tSZ data obtained with the NIKA camera, which is a dual-band (150 and 260~GHz) instrument operated at the IRAM 30-meter telescope. We deproject jointly NIKA and \planck\ data to extract the electronic pressure distribution from the cluster core ($R \sim 0.02\, R_{500}$) to its outskirts ($R \sim 3\, R_{500}$) non-parametrically for the first time at intermediate redshift. The constraints on the resulting pressure profile allow us to reduce the relative uncertainty on the integrated Compton parameter by a factor of two compared to the \planck\ value. Combining the tSZ data and the deprojected electronic density profile from \xmm\ allows us to undertake a hydrostatic mass analysis, for which we study the impact of a spherical model assumption on the total mass estimate. We also investigate the radial temperature and entropy distributions. These data indicate that \psz\ is a massive ($M_{500} \sim 5.5 \times 10^{14}$ M$_{\odot}$) cool-core cluster. This work is part of a pilot study aiming at optimizing the treatment of the NIKA2 tSZ large program dedicated to the follow-up of SZ-discovered clusters at intermediate and high redshifts. (abridged)Comment: 16 pages, 10 figure

NIKA2: a mm camera for cluster cosmology

Galaxy clusters constitute a major cosmological probe. However, Planck 2015 results have shown a weak tension between CMB-derived and cluster-derived cosmological parameters. This tension might be due to poor knowledge of the cluster mass and observable relationship. As for now, arcmin resolution Sunyaev-Zeldovich (SZ) observations ({\it e.g.} SPT, ACT and Planck) only allowed detailed studies of the intra cluster medium for low redshift clusters ($z0.5$) high resolution and high sensitivity SZ observations are needed. With both a wide field of view (6.5 arcmin) and a high angular resolution (17.7 and 11.2 arcsec at 150 and 260 GHz), the NIKA2 camera installed at the IRAM 30-m telescope (Pico Veleta, Spain) is particularly well adapted for these observations. The NIKA2 SZ observation program will map a large sample of clusters (50) at redshifts between 0.5 and 0.9. As a pilot study for NIKA2, several clusters of galaxies have been observed with the pathfinder, NIKA, at the IRAM 30-m telescope to cover the various configurations and observation conditions expected for NIKA2.

High angular resolution Sunyaev-Zel'dovich observations of MACS J1423.8+2404 with NIKA: Multiwavelength analysis

The prototype of the NIKA2 camera, NIKA, is an instrument operating at the IRAM 30-m telescope, which can observe simultaneously at 150 and 260GHz. One of the main goals of NIKA2 is to measure the pressure distribution in galaxy clusters at high resolution using the thermal SZ (tSZ) effect. Such observations have already proved to be an excellent probe of cluster pressure distributions even at high redshifts. However, an important fraction of clusters host submm and/or radio point sources, which can significantly affect the reconstructed signal. Here we report on <20" resolution observations at 150 and 260GHz of the cluster MACSJ1424, which hosts both radio and submm point sources. We examine the morphology of the tSZ signal and compare it to other datasets. The NIKA data are combined with Herschel satellite data to study the SED of the submm point source contaminants. We then perform a joint reconstruction of the intracluster medium (ICM) electronic pressure and density by combining NIKA, Planck, XMM-Newton, and Chandra data, focusing on the impact of the radio and submm sources on the reconstructed pressure profile. We find that large-scale pressure distribution is unaffected by the point sources because of the resolved nature of the NIKA observations. The reconstructed pressure in the inner region is slightly higher when the contribution of point sources are removed. We show that it is not possible to set strong constraints on the central pressure distribution without accurately removing these contaminants. The comparison with X-ray only data shows good agreement for the pressure, temperature, and entropy profiles, which all indicate that MACSJ1424 is a dynamically relaxed cool core system. The present observations illustrate the possibility of measuring these quantities with a relatively small integration time, even at high redshift and without X-ray spectroscopy.Comment: 15 pages, 17 figures, submitted to A&

High-resolution tSZ cartography of clusters of galaxies with NIKA at the IRAM 30-m telescope

The thermal Sunyaev-Zeldovich effect (tSZ) is a powerful probe to study clusters of galaxies and is complementary with respect to X-ray, lensing or optical observations. Previous arcmin resolution tSZ observations ({\it e.g.} SPT, ACT and Planck) only enabled detailed studies of the intra-cluster medium morphology for low redshift clusters ($z < 0.2$). Thus, the development of precision cosmology with clusters requires high angular resolution observations to extend the understanding of galaxy cluster towards high redshift. NIKA2 is a wide-field (6.5 arcmin field of view) dual-band camera, operated at $100 \ {\rm mK}$ and containing $\sim 3300$ KID (Kinetic Inductance Detectors), designed to observe the millimeter sky at 150 and 260 GHz, with an angular resolution of 18 and 12 arcsec respectively. The NIKA2 camera has been installed on the IRAM 30-m telescope (Pico Veleta, Spain) in September 2015. The NIKA2 tSZ observation program will allow us to observe a large sample of clusters (50) at redshift ranging between 0.5 and 1. As a pathfinder for NIKA2, several clusters of galaxies have been observed at the IRAM 30-m telescope with the NIKA prototype to cover the various configurations and observation conditions expected for NIKA2.Comment: Proceedings of the 28th Texas Symposium on Relativistic Astrophysics, Geneva, Switzerland, December 13-18, 201

Nika2: A mm camera for cluster cosmology

Galaxy clusters constitute a major cosmological probe. However, Planck 2015 results have shown a weak tension between CMB-derived and cluster-derived cosmological parameters. This tension might be due to poor knowledge of the cluster mass and observable relationship. As for now, arcmin resolution Sunyaev-Zeldovich (SZ) observations (e.g. SPT, ACT and Planck) only allowed detailed studies of the intra cluster medium for low redshift clusters (z 0:5) high resolution and high sensitivity SZ observations are needed. With both a wide field of view (6.5 arcmin) and a high angular resolution (17.7 and 11.2 arcsec at 150 and 260 GHz), the NIKA2 camera installed at the IRAM 30-m telescope (Pico Veleta, Spain) is particularly well adapted for these observations. The NIKA2 SZ observation program will map a large sample of clusters (50) at redshifts between 0.5 and 0.9. As a pilot study for NIKA2, several clusters of galaxies have been observed with the pathfinder, NIKA, at the IRAM 30-m telescope to cover the various configurations and observation conditions expected for NIKA2

First polarised light with the NIKA camera

NIKA is a dual-band camera operating with 315 frequency multiplexed LEKIDs cooled at 100 mK. NIKA is designed to observe the sky in intensity and polarisation at 150 and 260 GHz from the IRAM 30-m telescope. It is a test-bench for the final NIKA2 camera. The incoming linear polarisation is modulated at four times the mechanical rotation frequency by a warm rotating multi-layer Half Wave Plate. Then, the signal is analysed by a wire grid and finally absorbed by the LEKIDs. The small time constant (< 1ms ) of the LEKID detectors combined with the modulation of the HWP enables the quasi-simultaneous measurement of the three Stokes parameters I, Q, U, representing linear polarisation. In this paper we present results of recent observational campaigns demonstrating the good performance of NIKA in detecting polarisation at mm wavelength.Comment: 7 pages, Proceeding for Journal of Low Temperature Physic

The NIKA2 instrument, a dual-band kilopixel KID array for millimetric astronomy

NIKA2 (New IRAM KID Array 2) is a camera dedicated to millimeter wave astronomy based upon kilopixel arrays of Kinetic Inductance Detectors (KID). The pathfinder instrument, NIKA, has already shown state-of-the-art detector performance. NIKA2 builds upon this experience but goes one step further, increasing the total pixel count by a factor $\sim$10 while maintaining the same per pixel performance. For the next decade, this camera will be the resident photometric instrument of the Institut de Radio Astronomie Millimetrique (IRAM) 30m telescope in Sierra Nevada (Spain). In this paper we give an overview of the main components of NIKA2, and describe the achieved detector performance. The camera has been permanently installed at the IRAM 30m telescope in October 2015. It will be made accessible to the scientific community at the end of 2016, after a one-year commissioning period. When this happens, NIKA2 will become a fundamental tool for astronomers worldwide.Comment: Proceedings of the 16th Low Temperature Detectors workshop. To be published in the Journal of Low Temperature Physics. 8 pages, 4 figures, 1 tabl