Dynamic spatiotemporal coordination of neural stem cell fate decisions occurs through local feedback in the adult vertebrate brain

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Biomonitoring of air pollution as exemplified by recent IAEA programs

Biomonitoring is an appropriate tool for assessing the levels of atmospheric pollution, having several advantages compared with the use of direct measurements of contaminants (e.g., in airborne particulate matter, atmospheric deposition, precipitation), related primarily to the permanent and common occurrence of the chosen organisms in the field, the ease of sampling, and trace element accumulation. Furthermore, biomonitors may provide a measure of integrated exposure over an extended period of time and are present in remote areas and no expensive technical equipment is involved in collecting them. They accumulate contaminants over the exposure time and concentrate them, thus facilitating analytical measurements. Based on largescale biomonitoring surveys, polluted areas can be identified, and by applying appropriate statistical tools, information can be obtained on the type of pollution sources and on the transboundary transport of atmospheric pollutants. The International Atomic Energy Agency is including the research on biomonitors in its projects on health-related environmental studies. Biomonitoring activities from several coordinated research projects on air pollution are presented, and results from an international workshop are discussed. In addition, activities in supporting improvement quality in the participating laboratories are outlined.     

Real-time detection of mAb aggregates in an integrated downstream process

The implementation of continuous processing in the biopharmaceutical industry is hindered by the scarcity of process analytical technologies (PAT). To monitor and control a continuous process, PAT tools will be crucial to measure real-time product quality attributes such as protein aggregation. Miniaturizing these analytical techniques can increase measurement speed and enable faster decision-making. A fluorescent dye (FD)-based miniaturized sensor has previously been developed: a zigzag microchannel which mixes two streams under 30 s. Bis-ANS and CCVJ, two established FDs, were employed in this micromixer to detect aggregation of the biopharmaceutical monoclonal antibody (mAb). Both FDs were able to robustly detect aggregation levels starting at 2.5%. However, the real-time measurement provided by the microfluidic sensor still needs to be implemented and assessed in an integrated continuous downstream process. In this work, the micromixer is implemented in a lab-scale integrated system for the purification of mAbs, established in an ÄKTA™ unit. A viral inactivation and two polishing steps were reproduced, sending a sample of the product pool after each phase directly to the microfluidic sensor for aggregate detection. An additional UV sensor was connected after the micromixer and an increase in its signal would indicate that aggregates were present in the sample. The at-line miniaturized PAT tool provides a fast aggregation measurement, under 10 min, enabling better process understanding and control.     

Online deployment of an O-PLS model for dielectric spectroscopy-based inline monitoring of viable cell concentrations in Chinese hamster ovary cell perfusion cultivations

Viable cell concentration (VCC) is an essential parameter that is required to support the efficient cultivation of mammalian cells. Although commonly determined using at-line or off-line analytics, in-line capacitance measurements represent a suitable alternative method for the determination of VCC. In addition, these latter efforts are complimentary with the Food and Drug Administration's initiative for process analytical technologies (PATs). However, current applications for online determination of the VCC often rely on single frequency measurements and corresponding linear regression models. It has been reported that this may be insufficient for application at all stages of a mammalian cell culture processes due to changes in multiple cell parameters over time. Alternatively, dielectric spectroscopy, measuring capacitance at multiple frequencies, in combination with multivariate mathematical models, has proven to be more robust. However, this has only been applied for retrospective data analysis. Here, we present the implementation of an O-PLS model for the online processing of multifrequency capacitance signals and the on-the-fly integration of the models’ VCC results into a supervisory control and data acquisition (SCADA) system commonly used for cultivation observation and control. This system was evaluated using a Chinese hamster ovary (CHO) cell perfusion process.     

The in-line measurement of plant cell biomass using radio frequency impedance spectroscopy as a component of process analytical technology

Radio frequency impedance spectroscopy (RFIS) is a robust method for the determination of cell biomass during fermentation. RFIS allows non-invasive in-line monitoring of the passive electrical properties of cells in suspension and can distinguish between living and dead cells based on their distinct behavior in an applied radio frequency field. We used continuous in situ RFIS to monitor batch-cultivated plant suspension cell cultures in stirred-tank bioreactors and compared the in-line data to conventional off-line measurements. RFIS-based analysis was more rapid and more accurate than conventional biomass determination, and was sensitive to changes in cell viability. The higher resolution of the in-line measurement revealed subtle changes in cell growth which were not accessible using conventional methods. Thus, RFIS is well suited for correlating such changes with intracellular states and product accumulation, providing unique opportunities for employing systems biotechnology and process analytical technology approaches to increase product yield and quality.     

Uncertainty Estimation in a Remediation Process: A Case Study

A case study concerning the reclamation of a brownfield for residential purposes is presented. Because a high contamination of Polycyclic Aromatic Hydrocarbons (PAHs) was detected in soil, a remediation process is necessary. Bench-scale treat-ability tests were carried out in order to evaluate performances of some remediation technologies on this specific matrix, and particularly for the removal of high-molecular-weight PAHs. Biodegradation studies allowed the evaluation of PAH abatement in a slurry phase treatment, the amount of abiotic losses and the effect of macronutri-ents and bioaugmentation on the removal efficiency. The experimental study was performed in compliance with a Quality System, based on ISO 9001:2000 and ISO/ EC 17025:1999; validation of the analytical method provided the expanded uncertainty of the removal efficiency, varying from nearly 13 to 21%, depending on the compound considered. Experimental results showed a high removal efficiency for all PAHs; the addition of nitrogen and phosphorous increased the removal rate and the efficiency for high-molecular-weight PAHs, whereas no remarkable differences were observed in total (abiotic + biotic) removal of light compounds.