A typical application of SEAMLESS-IF at macro level: a trade liberalization scenario applied to the EU

Agricultural and Food Policy, Environmental Economics and Policy, Land Economics/Use, Production Economics,

Application of multivariate data analysis in the monitoring and control of mammalian cell processes

High throughput (HT) methodologies are increasingly being adopted for bioprocess development activities. However, often the large quantities of data generated from such studies as well as from historical batch records are not fully harnessed for their potential insights. Multivariate data analysis (MVDA) is a well-known technique that can reduce the dimensionality of large data sets and help generate useful correlations of typical process behaviour and determine root causes of process deviations. This work focuses on the application of various MVDA techniques and data mining tools to investigate the impact of process variations on the critical quality attributes (CQAs) of mammalian cell processes. Three separate projects investigating the application of these techniques were explored. The first focuses on understanding the impact of cell culture operation, generation number and initial conditions on final titre and impurity levels. A systematic methodology was applied to analyse HT data generated in a Design of Experiment (DOE) study using the ambr® 48 (advanced micro-bioreactor) system. The project applied multiple MVDA techniques including Principle Component Analysis (PCA) and Partial Least Squares (PLS) to successfully identify the key critical process measurements impacting on the target antibody concentration and host cell protein (HCP) levels at harvest. A similar approach was adopted in the second project that investigated the influence of manufacturing beyond standard operating conditions on the product-related CQAs of a mammalian cell process. The insights from the MVDA allowed the modification of the control limits of key process parameters to be redefined with confidence. The final project focuses on the development of an advanced glucose control strategy for a high titre mammalian cell line. The project aims to predict the on-line glucose concentration through correlations developed between the available on-line process measurements and the off-line metabolic profiles. The predicted glucose concentration is then used to manipulate the substrate feed rate to control the glucose concentration at a desired set-point. Better control of the glucose concentration aims to further increase titre production as well as potentially reducing unwanted post-translation modifications (PTMs) in susceptible cell lines. These techniques can be combined with the application of Process Analytical Technologies (PAT) and Quality by Design (QbD) allowing for the development of more efficient and better controlled processes

Reliability and validity of the Brief Illness Perception Questionnaire (B-IPQ) in individuals with a recently acquired spinal cord injury

Objective To assess the reliability and validity of the Brief Illness Perception Questionnaire (B-IPQ) and possible subscales, and to interpret Brief Illness Perception Questionnaire (B-IPQ) total scores, in individuals with a spinal cord injury. Design Cross-sectional Setting Seven Dutch rehabilitation centres Subjects Individuals with a recently acquired traumatic or non-traumatic spinal cord injury at the start of inpatient spinal cord injury rehabilitation (N = 270). Main measure The Brief Illness Perception Questionnaire (B-IPQ) consists of eight items on an individual's cognitive and emotional representation of one's health conditions. Principal component analysis was performed to identify possible Brief Illness Perception Questionnaire (B-IPQ) subscales. Validity was assessed by testing hypotheses on correlations between the Brief Illness Perception Questionnaire (B-IPQ) and other measures. Cut-off points of the Brief Illness Perception Questionnaire (B-IPQ) total score were determined. Results Mean (SD) age of participants was 60.1 (16.5) years, 188 (71%) were male, and 119 (44%) had tetraplegia. Three potential subscales were revealed. Cronbach's alpha was acceptable for only one subscale. This subscale was named 'consequences' and included the items 'consequences', 'symptom burden', 'concern', and 'emotions'. The Brief Illness Perception Questionnaire (B-IPQ) total and the consequence subscale showed the expected strong correlations (>.50) with symptoms of anxiety and depression. Mean (SD) scores were 40.9 (12.3) on the 8-item Brief Illness Perception Questionnaire (B-IPQ) (range 0-80) and 25.1 (8.1) on the consequences subscale (range 0-40). Cut-off points for the Brief Illness Perception Questionnaire (B-IPQ) total score were determined as follows: = 50 indicating high experienced threat. Conclusion The Brief Illness Perception Questionnaire (B-IPQ) total and consequences subscale seem applicable in individuals with a spinal cord injury in the rehabilitation practice and research

Post-traumatic stress disorder symptoms and pain intensity in persons with spinal cord injury

Study design Cross-sectional. Objectives To examine the association between post-traumatic stress disorder (PTSD) symptoms and pain intensity, taking symptoms of anxiety and depression into account within persons with spinal cord injury (SCI). Setting Persons with SCI, who visited a Dutch rehabilitation centre between 2005 and 2010, were invited to complete a survey. Methods PTSD symptoms were measured with the Trauma Screening Questionnaire (TSQ), pain intensity with an 11-point Numerical Rating Scale (NRS), and symptoms of anxiety and depression with the Hospital Anxiety and Depression Scale (HADS). To determine associations between PTSD symptoms and pain intensity, linear regression analyses were performed. Confounding variables representing anxiety and depression were added to the final model. Results In total, 175 participants (55.8% traumatic, 29.1% complete) were included (response rate of 31.7%). Of them, 11.4% had clinically relevant symptoms of probable PTSD (TSQ score >= 6) 69.8% experienced moderate to severe pain levels (NRS >= 4), 14.9% had symptoms of anxiety and 20.8% symptoms of depression (HADS scores >= 11). Levels of PTSD symptoms were strongly associated with symptoms of anxiety (0.54) and depression (0.49). Bivariate analyses showed a moderate significant association (0.30) between PTSD symptoms and pain intensity. This association became small (0.10) when anxiety and depression comorbidity were factored into the final regression model. Conclusions No independent association between PTSD symptoms and pain intensity was shown when adjusted for anxiety and depression. Results of this study suggest the usefulness of screening for PTSD in persons with SCI (regardless of injury cause or type/level) who score high on symptoms of anxiety/depression

Functionalized micro-capillary film for the rapid at-line analysis of IgG aggregates in a cell culture bioreactor.

A micro-capillary film has been developed that offers the potential for an at-line analytical tool for rapid aggregate analysis during biopharmaceutical antibody production. A non-porous walled micro-capillary film (NMCF) with cation exchange functionality was demonstrated to act as a chromatography medium that could be operated with high linear fluid velocities and was highly resistant to blockage by entrained particulates, including cells. The NMCF containing 19 parallel microcapillaries was prepared using a melt extrusion process from poly(ethylene-vinyl alcohol) copolymer (EVOH). The NMCF-EVOH was modified to have cation-exchange functionality (NMCF-EVOH-SP) and shown to differentially bind monomer and aggregated species of IgG antibody directly from a bioreactor. The use of NMCF-EVOH-SP to quantify aggregate concentrations in monoclonal antibody preparations in less than 20 minutes was demonstrated.The authors would like to thank the EPSRC for the provision of a CASE Award. This study was sponsored by MedImmune, the global biologics R&D arm of AstraZeneca.This is the final version of the article. It first appeared from Taylor & Francis via http://dx.doi.org/10.1080/19420862.2015.106536

Is disturbed clearance of apoptotic keratinocytes responsible for UVB-induced inflammatory skin lesions in systemic lupus erythematosus?

Apoptotic cells are thought to play an essential role in the pathogenesis of systemic lupus erythematosus (SLE). We hypothesise that delayed or altered clearance of apoptotic cells after UV irradiation will lead to inflammation in the skin of SLE patients. Fifteen SLE patients and 13 controls were irradiated with two minimal erythemal doses (MEDs) of ultraviolet B light (UVB). Subsequently, skin biopsies were analysed (immuno)histologically, over 10 days, for numbers of apoptotic cells, T cells, macrophages, and deposition of immunoglobulin and complement. Additionally, to compare results with cutaneous lesions of SLE patients, 20 biopsies of lupus erythematosus (LE) skin lesions were analysed morphologically for apoptotic cells and infiltrate. Clearance rate of apoptotic cells after irradiation did not differ between patients and controls. Influx of macrophages in dermal and epidermal layers was significantly increased in patients compared with controls. Five out of 15 patients developed a dermal infiltrate that was associated with increased epidermal influx of T cells and macrophages but not with numbers of apoptotic cells or epidermal deposition of immunoglobulins. Macrophages were ingesting multiple apoptotic bodies. Inflammatory lesions in these patients were localised near accumulations of apoptotic keratinocytes similar as was seen in the majority of LE skin lesions. In vivo clearance rate of apoptotic cells is comparable between SLE patients and controls. However, the presence of inflammatory lesions in the vicinity of apoptotic cells, as observed both in UVB-induced and in LE skin lesions in SLE patients, suggests that these lesions result from an inflammatory clearance of apoptotic cells

Superplume mantle tracked isotopically the length of Africa from the Indian Ocean to the Red Sea

Seismological findings show a complex scenario of plume upwellings from a deep thermo-chemical anomaly (superplume) beneath the East African Rift System (EARS). It is unclear if these geophysical observations represent a true picture of the superplume and its influence on magmatism along the EARS. Thus, it is essential to find a geochemical tracer to establish where upwellings are connected to the deep-seated thermo-chemical anomaly. Here we identify a unique non-volatile superplume isotopic signature (‘C’) in the youngest (after 10 Ma) phase of widespread EARS rift-related magmatism where it extends into the Indian Ocean and the Red Sea. This is the first sound evidence that the superplume influences the EARS far from the low seismic velocities in the magma-rich northern half. Our finding shows for the first time that superplume mantle exists beneath the rift the length of Africa from the Red Sea to the Indian Ocean offshore southern Mozambiqu

On-ward participation of a hospital pharmacist in a Dutch intensive care unit reduces prescribing errors and related patient harm: an intervention study

Introduction: Patients admitted to an intensive care unit (ICU) are at high risk for prescribing errors and related adverse drug events (ADEs). An effective intervention to decrease this risk, based on studies conducted mainly in North America, is on-ward participation of a clinical pharmacist in an ICU team. As the Dutch Healthcare System is organized differently and the on-ward role of hospital pharmacists in Dutch ICU teams is not well established, we conducted an intervention study to investigate whether participation of a hospital pharmacist can also be an effective approach in reducing prescribing errors and related patient harm (preventable ADEs) in this specific setting. Methods: A prospective study compared a baseline period with an intervention period. During the intervention period, an ICU hospital pharmacist reviewed medication orders for patients admitted to the ICU, noted issues related to prescribing, formulated recommendations and discussed those during patient review meetings with the attending ICU physicians. Prescribing issues were scored as prescribing errors when consensus was reached between the ICU hospital pharmacist and ICU physicians. Results: During the 8.5-month study period, medication orders for 1,173 patients were reviewed. The ICU hospital pharmacist made a total of 659 recommendations. During the intervention period, the rate of consensus between the ICU hospital pharmacist and ICU physicians was 74%. The incidence of prescribing errors during the intervention period was significantly lower than during the baseline period: 62.5 per 1,000 monitored patient-days versus 190.5 per 1,000 monitored patient-days, respectively (P < 0.001). Preventable ADEs (patient harm, National Coordinating Council for Medication Error Reporting and Prevention severity categories E and F) were reduced from 4.0 per 1,000 monitored patient-days during the baseline period to 1.0 per 1,000 monitored patient-days during the intervention period (P = 0.25). Per monitored patient-day, the intervention itself cost (sic)3, but might have saved (sic)26 to (sic)40 by preventing ADEs. Conclusions: On-ward participation of a hospital pharmacist in a Dutch ICU was associated with significant reductions in prescribing errors and related patient harm (preventable ADEs) at acceptable costs per monitored patient-da

In Vitro Cytochrome P450 Formation of a Mono-Hydroxylated Metabolite of Zearalenone Exhibiting Estrogenic Activities: Possible Occurrence of This Metabolite in Vivo

The mycoestrogen zearalenone (ZEN), as well as its reduced metabolites, which belong to the endocrine disruptor bio-molecule family, are substrates for various enzymes involved in steroid metabolism. In addition to its reduction by the steroid dehydrogenase pathway, ZEN also interacts with hepatic detoxification enzymes, which convert it into hydroxylated metabolites (OH-ZEN). Due to their structures to that of estradiol, ZEN and its derived metabolites bind to the estrogen receptors and are involved in endocrinal perturbations and are possibly associated with estrogen-dependent cancers. The primary aim of this present study was to identify the enzymatic cytochrome P450 isoforms responsible for the formation of the most abundant OH-ZEN. We thus studied its in vitro formation using hepatic microsomes in a range of animal model systems including man. OH-ZEN was also recovered in liver and urine of rats treated orally with ZEN. Finally we compared the activity of ZEN and its active metabolites (α-ZAL and OH-ZEN) on estrogen receptors using HeLa ER-α and ER-β reporter cell lines as reporters. OH-ZEN estrogenic activities were revealed to be limited and not as significant as those of ZEN or α-ZAL

Breast cancer oestrogen independence mediated by BCAR1 or BCAR3 genes is transmitted through mechanisms distinct from the oestrogen receptor signalling pathway or the epidermal growth factor receptor signalling pathway

INTRODUCTION: Tamoxifen is effective for endocrine treatment of oestrogen receptor-positive breast cancers but ultimately fails due to the development of resistance. A functional screen in human breast cancer cells identified two BCAR genes causing oestrogen-independent proliferation. The BCAR1 and BCAR3 genes both encode components of intracellular signal transduction, but their direct effect on breast cancer cell proliferation is not known. The aim of this study was to investigate the growth control mediated by these BCAR genes by gene expression profiling. METHODS: We have measured the expression changes induced by overexpression of the BCAR1 or BCAR3 gene in ZR-75-1 cells and have made direct comparisons with the expression changes after cell stimulation with oestrogen or epidermal growth factor (EGF). A comparison with published gene expression data of cell models and breast tumours is made. RESULTS: Relatively few changes in gene expression were detected in the BCAR-transfected cells, in comparison with the extensive and distinct differences in gene expression induced by oestrogen or EGF. Both BCAR1 and BCAR3 regulate discrete sets of genes in these ZR-75-1-derived cells, indicating that the proliferation signalling proceeds along distinct pathways. Oestrogen-regulated genes in our cell model showed general concordance with reported data of cell models and gene expression association with oestrogen receptor status of breast tumours. CONCLUSIONS: The direct comparison of the expression profiles of BCAR transfectants and oestrogen or EGF-stimulated cells strongly suggests that anti-oestrogen-resistant cell proliferation is not caused by alternative activation of the oestrogen receptor or by the epidermal growth factor receptor signalling pathway