Sanofi in-house medium: Exceeding expectations in cell line development as an alternative to commercially available basal medium

Our current CHO cell line development process has used commercially supplied media for all steps from post-transfection recovery through scale-up and screening of clones. Media alternatives have been developed in house to improve cell culture performance, decrease media cost, and enable modification by Sanofi. A current version of the basal medium has been utilized for cell culture production processes for some early stage development programs. With the goal of utilizing the same media for cell line development and cell culture production, we evaluated this basal medium in different steps of the cell line generation process, with the current (commercial) medium used as the control. Overall, the results demonstrated that the in-house medium met or exceeded the performance of the commercial medium in the cell line development workflow. To assess the in-house basal medium for post-transfection methotrexate (MTX) selection, cell growth and viability were measured during MTX selection, and recombinant protein titers of the MTX-selected pools were assessed. Results show that protein production is comparable between pools selected in the in-house medium versus the control. In addition, use of the in-house medium improved cell growth during selection and decreased the selection timeline by a week. A subsequent MTX kill curve study using the in-house medium showed excellent recovery of transfected cells, even at the higher MTX concentrations used, and high protein titers with less time in selection. Next, the medium was evaluated for single cell cloning in a process using flow cytometry cell sorting to deposit one cell per well in 96-well plates. The initial experiments revealed a minor limitation of the in-house medium for single cell cloning and, based on these results, a cloning version of the basal medium was prepared and tested. Several experiments demonstrated that the cloning version of the in-house medium supported robust cell growth and cloning efficiency that is similar to or better than that obtained with the control. Further studies showed that, for both pools and clones in batch cultures, the in-house medium supports more rapid growth rates and higher cell densities. Experimental results to be presented demonstrate success of the in-house medium in the MTX selection, single cell cloning and outgrowth, and pool and clone screening steps of the cell line development workflow. Furthermore, the advantages of using the in-house medium for pre-master cell banking will be discussed. Proof of concept work from recent cell line generation projects will further demonstrate the utility and advantage of using our in-house medium in the cell line development workflow for new preclinical development programs

Keeping it liminal. The Mondiali Antirazzisti (Anti-racist World Cup) as a multifocal interaction ritual

© 2013 Taylor & Francis. This paper examines how social mixing and celebration of diversity can be enabled through sports festivals marked by their carnivalesque atmosphere. Our analysis draws on a longitudinal ethnographic study of the Mondiali Antirazzisti (Anti-racist World Cup), a non-competitive football tournament and intercultural festival featuring the yearly participation of hardcore football fans (ultras), migrant groups, third-sector associations and other informal groups. We consider how the multifocal ritual form of the event helps to create a liminal space in which discrimination and stereotypes can be temporarily challenged. The sources of collective effervescence are multiplied by placing sport games within a wider range of other leisure and cultural activities, thus promoting internal diversity and the inclusion of outsiders. Additionally, social boundaries are also blurred by not emphasising the competitive dimension of the sporting activities, making sporting categorisations more fluid, and breaking down the separation between protagonists and spectators. Nonetheless, considering the transient character of liminality, we also investigate problems and limitations implied by the pursuit of these objectives. It is concluded that, despite a certain degree of self-referentiality, the festival fosters the spreading of anti-discriminatory cultures by enhancing the participants’ reflexivity and feeding their commitment in generating spin-off activities in different local contexts

Engaging stakeholders in rehabilitation research: a scoping review of strategies used in partnerships and evaluation of impacts

Abstract: Purpose: To describe how stakeholder engagement has been undertaken and evaluated in rehabilitation research. Methods: A scoping review of the scientific literature using five search strategies. Quantitative and qualitative analyses using extracted data. Interpretation of results was iteratively discussed within the team, which included a parent stakeholder. Results: Searches identified 101 candidate papers; 28 were read in full to assess eligibility and 19 were included in the review. People with disabilities and their families were more frequently involved compared to other stakeholders. Stakeholders were often involved in planning and evaluating service delivery. A key issue was identifying stakeholders; strategies used to support their involvement included creating committees, organizing meetings, clarifying roles and offering training. Communication, power sharing and resources influenced how stakeholders could be engaged in the research. Perceived outcomes of stakeholder engagement included the creation of partnerships, facilitating the research process and the application of the results, and empowering stakeholders. Stakeholder engagement outcomes were rarely formally evaluated. Conclusions: There is a great interest in rehabilitation to engage stakeholders in the research process. However, further evidence is needed to identify effective strategies for meaningful stakeholder engagement that leads to more useful research that positively impacts practice. Implications for Rehabilitation Using several strategies to engage various stakeholders throughout the research process is thought to increase the quality of the research and the rehabilitation process by developing proposals and programs responding better to their needs. Engagement strategies need to be better reported and evaluated in the literature. Engagement facilitate uptake of research findings by increasing stakeholders' awareness of the evidence, the resources available and their own ability to act upon a situation. Factors influencing opportunities for stakeholder engagement need to be better understood

Population pharmacokinetics of ciprofloxacin in neonates and young infants less than 3 months age

Ciprofloxacin is used in neonates with suspected or documented Gram-negative serious infections. Currently, its use is off-label partly because of lack of pharmacokinetic studies. Within the FP7 EU project TINN (Treat Infection in NeoNates), our aim was to evaluate the population pharmacokinetics of ciprofloxacin in neonates and young infants \u3c3 months of age and define the appropriate dose in order to optimize ciprofloxacin treatment in this vulnerable population. Blood samples were collected from neonates treated with ciprofloxacin and concentrations were quantified by high-pressure liquid chromatography-mass spectrometry. Population pharmacokinetic analysis was performed using NONMEM software. The data from 60 newborn infants (postmenstrual age [PMA] range, 24.9 to 47.9 weeks) were available for population pharmacokinetic analysis. A two-compartment model with first-order elimination showed the best fit with the data. A covariate analysis identified that gestational age, postnatal age, current weight, serum creatinine concentration, and use of inotropes had a significant impact on ciprofloxacin pharmacokinetics. Monte Carlo simulation demonstrated that 90% of hypothetical newborns with a PMA of \u3c34 weeks treated with 7.5 mg/kg twice daily and 84% of newborns with a PMA ≥34 weeks and young infants receiving 12.5 mg/kg twice daily would reach the AUC/MIC target of 125, using the standard EUCAST MIC susceptibility breakpoint of 0.5 mg/liter. The associated risks of overdose for the proposed dosing regimen were \u3c8%. The population pharmacokinetics of ciprofloxacin was evaluated in neonates and young infants \u3c3 months old, and a dosing regimen was established based on simulation

Morphological characteristics of motor neurons do not determine their relative susceptibility to degeneration in a mouse model of severe spinal muscular atrophy

Spinal muscular atrophy (SMA) is a leading genetic cause of infant mortality, resulting primarily from the degeneration and loss of lower motor neurons. Studies using mouse models of SMA have revealed widespread heterogeneity in the susceptibility of individual motor neurons to neurodegeneration, but the underlying reasons remain unclear. Data from related motor neuron diseases, such as amyotrophic lateral sclerosis (ALS), suggest that morphological properties of motor neurons may regulate susceptibility: in ALS larger motor units innervating fast-twitch muscles degenerate first. We therefore set out to determine whether intrinsic morphological characteristics of motor neurons influenced their relative vulnerability to SMA. Motor neuron vulnerability was mapped across 10 muscle groups in SMA mice. Neither the position of the muscle in the body, nor the fibre type of the muscle innervated, influenced susceptibility. Morphological properties of vulnerable and disease-resistant motor neurons were then determined from single motor units reconstructed in Thy.1-YFP-H mice. None of the parameters we investigated in healthy young adult mice - including motor unit size, motor unit arbor length, branching patterns, motor endplate size, developmental pruning and numbers of terminal Schwann cells at neuromuscular junctions - correlated with vulnerability. We conclude that morphological characteristics of motor neurons are not a major determinant of disease-susceptibility in SMA, in stark contrast to related forms of motor neuron disease such as ALS. This suggests that subtle molecular differences between motor neurons, or extrinsic factors arising from other cell types, are more likely to determine relative susceptibility in SMA

Adverse Life Trajectories Are a Risk Factor for SARS-CoV-2 IgA Seropositivity.

Asymptomatic individuals, called "silent spreaders" spread SARS-CoV-2 efficiently and have complicated control of the ongoing COVID-19 pandemic. As seen in previous influenza pandemics, socioeconomic and life-trajectory factors are important in disease progression and outcome. The demographics of the asymptomatic SARS-CoV-2 carriers are unknown. We used the CON-VINCE cohort of healthy, asymptomatic, and oligosymptomatic individuals that is statistically representative of the overall population of Luxembourg for age, gender, and residency to characterise this population. Gender (male), not smoking, and exposure to early-life or adult traumatic experiences increased the risk of IgA seropositivity, and the risk associated with early-life exposure was a dose-dependent metric, while some other known comorbidities of active COVID-19 do not impact it. As prior exposure to adversity is associated with negative psychobiological reactions to external stressors, we recorded psychological wellbeing during the study period. Exposure to traumatic events or concurrent autoimmune or rheumatic disease were associated with a worse evolution of anxiety and depressive symptoms throughout the lockdown period. The unique demographic profile of the "silent spreaders" highlights the role that the early-life period plays in determining our lifelong health trajectory and provides evidence that the developmental origins of health and disease is applicable to infectious diseases

Colloids in a bacterial bath: simulations and experiments

We present a joint experimental and computational study of the effect of bacterial motion on micron-scale colloids contained in a two-dimensional suspension of Bacillus subtilis. With respect to previous work using E. coli, here we introduce a novel experimental set-up that allows us to realise a two-dimensional bacterial suspension insensitive to either evaporation or fluid flow. By analysing the mean square displacements of both bacteria and colloids, we confirm the existence of a crossover from super-diffusive behaviour at short time scales to normal diffusion at longer times. We also study the same two-dimensional system by means of numerical simulations, using a suspension of self-propelled dumbbells or the Vicsek model, which has been previously used to study the dynamics of active particles. Our numerical results obtained with both models are in broad agreement with the experimental trends, but only the dumbbell simulations can match the experimental data quantitatively. The level of agreement we find suggest that steric interactions due to collisions are important players in determining collective motion of the bacterial bath, and should complement hydrodynamic interactions in experiments

Forgotten Plotlanders: Learning from the survival of lost informal housing in the UK.

Colin Ward’s discourses on the arcadian landscape of ‘plotlander’ housing are unique documentations of the anarchistic birth, life, and death of the last informal housing communities in the UK. Today the forgotten history of ‘plotlander’ housing documented by Ward can be re-read in the context of both the apparently never-ending ‘housing crisis’ in the UK, and the increasing awareness of the potential value of learning from comparable informal housing from the Global South. This papers observations of a previously unknown and forgotten plotlander site offers a chance to begin a new conversation regarding the positive potential of informal and alternative housing models in the UK and wider Westernised world

Inter-rater reliability and stability of diagnoses of autism spectrum disorder in children identified through screening at a very young age

To examine the inter-rater reliability and stability of autism spectrum disorder (ASD) diagnoses made at a very early age in children identified through a screening procedure around 14 months of age. In a prospective design, preschoolers were recruited from a screening study for ASD. The inter-rater reliability of the diagnosis of ASD was measured through an independent assessment of a randomly selected subsample of 38 patients by two other psychiatrists. The diagnoses at 23 months and 42 months of 131 patients, based on the clinical assessment and the diagnostic classifications of standardised instruments, were compared to evaluate stability of the diagnosis of ASD. Inter-rater reliability on a diagnosis of ASD versus non-ASD at 23 months was 87% with a weighted κ of 0.74 (SE 0.11). The stability of the different diagnoses in the autism spectrum was 63% for autistic disorder, 54% for pervasive developmental disorder, not otherwise specified (PDD-NOS), and 91% for the whole category of ASD. Most diagnostic changes at 42 months were within the autism spectrum from autistic disorder to PDD-NOS and were mainly due to diminished symptom severity. Children who moved outside the ASD category at 42 months made significantly larger gains in cognitive and language skills than children with a stable ASD diagnosis. In conclusion, the inter-rater reliability and stability of the diagnoses of ASD established at 23 months in this population-based sample of very young children are good

Overestimation of Postpartum Depression Prevalence Based on a 5-item Version of the EPDS:Systematic Review and Individual Participant Data Meta-analysis

Objective:The Maternal Mental Health in Canada, 2018/2019, survey reported that 18% of 7,085 mothers who recently gave birth reported "feelings consistent with postpartum depression" based on scores >= 7 on a 5-item version of the Edinburgh Postpartum Depression Scale (EPDS-5). The EPDS-5 was designed as a screening questionnaire, not to classify disorders or estimate prevalence; the extent to which EPDS-5 results reflect depression prevalence is unknown. We investigated EPDS-5 >= 7 performance relative to major depression prevalence based on a validated diagnostic interview, the Structured Clinical Interview for DSM (SCID).Methods:We searched Medline, Medline In-Process & Other Non-Indexed Citations, PsycINFO, and the Web of Science Core Collection through June 2016 for studies with data sets with item response data to calculate EPDS-5 scores and that used the SCID to ascertain depression status. We conducted an individual participant data meta-analysis to estimate pooled percentage of EPDS-5 >= 7, pooled SCID major depression prevalence, and the pooled difference in prevalence.Results:A total of 3,958 participants from 19 primary studies were included. Pooled prevalence of SCID major depression was 9.2% (95% confidence interval [CI] 6.0% to 13.7%), pooled percentage of participants with EPDS-5 >= 7 was 16.2% (95% CI 10.7% to 23.8%), and pooled difference was 8.0% (95% CI 2.9% to 13.2%). In the 19 included studies, mean and median ratios of EPDS-5 to SCID prevalence were 2.1 and 1.4 times.Conclusions:Prevalence estimated based on EPDS-5 >= 7 appears to be substantially higher than the prevalence of major depression. Validated diagnostic interviews should be used to establish prevalence