Modelling concentration gradients in fed-batch cultivations of E. coli – towards the flexible design of scale-down experiments

The cover image is based on the Research Article Modelling concentration gradients in fed-batch cultivations of E. coli - towards the flexible design of scale-down experiments by Emmanuel Anane et al., DOI: 10.1002/jctb.5798 .

     

Functional Screen for microRNAs Suppressing Anchorage-Independent Growth in Human Cervical Cancer Cells

The progression of anchorage-dependent epithelial cells to anchorage-independent growth represents a critical hallmark of malignant transformation. Using an in vitro model of human papillomavirus (HPV)-induced transformation, we previously showed that acquisition of anchorage-independent growth is associated with marked (epi)genetic changes, including altered expression of microRNAs. However, the laborious nature of the conventional growth method in soft agar to measure this phenotype hampers a high-throughput analysis. We developed alternative functional screening methods using 96- and 384-well ultra-low attachment plates to systematically investigate microRNAs regulating anchorage-independent growth. SiHa cervical cancer cells were transfected with a microRNA mimic library (n = 2019) and evaluated for cell viability. We identified 84 microRNAs that consistently suppressed growth in three independent experiments. Further validation in three cell lines and comparison of growth in adherent and ultra-low attachment plates yielded 40 microRNAs that specifically reduced anchorage-independent growth. In conclusion, ultra-low attachment plates are a promising alternative for soft-agar assays to study anchorage-independent growth and are suitable for high-throughput functional screening. Anchorage independence suppressing microRNAs identified through our screen were successfully validated in three cell lines. These microRNAs may provide specific biomarkers for detecting and treating HPV-induced precancerous lesions progressing to invasive cancer, the most critical stage during cervical cancer development.     

A Longitudinal Study of Workspace Design for Knowledge Exploration and Exploitation in the Research and Development Process

In order to leverage organizational learning, scholars have already defined but are still discussing the interpretation of two different learning types, exploration and exploitation. Exploring new frontiers across knowledge domains and maintaining the balance with exploiting the existing knowledge is critical for the prosperity of an organization. The spatial dimension of organizational learning considers that proximity of employees has an influence on their learning activities, but from a rather macro perspective without taking workspace design into account. We account for these issues by examining the impact of workspace design on knowledge exploration and exploitation on the micro level at distinct stages along the value chain (i.e., the research, development and project market team unit) of Novartis, a pharmaceutical company. In a longitudinal study, employees of the three cases have been interviewed and observed over the course of three years, before and after workspace redesign. With the change from a cellular to an open workspace, employees become closer and highly visible to each other, which influences knowledge work. As the cases occurred sequentially in time, design principles were derived. The findings suggest that exploitation is supported by workspace design that leads to high proximity inducing faster feedback cycles and first‐hand information. Exploration, however, is supported by workspace design that leads to high visibility triggering more cross‐functional interactions and thereby the variability of knowledge. The later the stage in the research and development process, the higher the need for balanced learning activities. This balance is well reflected in a ‘multi‐space’ workspace consisting of shared meeting areas, quiet zones, central staircases and integrated laboratories and desk areas.     

Electric conductivity test for quality assessment of aromatic and medicinal plants after drying

For the production of essential oils and aromatics to be possible throughout the year, it is necessary to continuously supply the plant raw material to the industry or store the plant material for a designated time. However, to allow the plant material to remain in storage with a high quality, it is essential to reduce its water content by drying. This has motivated the oil extraction industries to demand a better yield and quality from the drying process and raw materials, necessitating fast and efficient evaluation parameters. Thus, the aim of this study was to develop an electric conductivity test for evaluating the quality of aromatic and medicinal plants and to quantify the essential oil yield of the dry vegetable material. The drying experiments were conducted at different temperatures and air velocities by altering the control of the drying conditions. Various aromatic plants with different plant mass, volume of deionized water, and temperature of the vegetal material were examined by the electric conductivity test. The qualities of the fresh and dry plants were compared through these tests and yields of essential oil. Increase in the drying air temperature influenced the increase in the dehydration rate of the aromatic and medicinal plants. Higher drying temperatures decreased the physical quality as evaluated by the electric conductivity test and essential oil yield of the aromatic and medicinal plants. The parameters for validation of the electric conductivity test methodology in aromatic and medicinal plants were defined as an exposure time of 33?h, 5?g of fresh plant material in 75?mL of deionized water, 1?g of dry plant material in 50?mL of deionized water, and temperature of 25°C. The electric conductivity test was demonstrated as an appropriate method to be used in the quality control of aromatic plants in essential oil extraction industries. The use of the electric conductivity test will enable the oil extraction industry to monitor the yield and quality of the essential oils extracted from the aromatic plants after drying and storage.     

Monoclonal Antibodies and Airway Diseases

Monoclonal antibodies, biologics, are a relatively new treatment option for severe chronic airway diseases, asthma, allergic rhinitis, and chronic rhinosinusitis (CRS). In this review, we focus on the physiological and pathomechanisms of monoclonal antibodies, and we present recent study results regarding their use as a therapeutic option against severe airway diseases. Airway mucosa acts as a relative barrier, modulating antigenic stimulation and responding to environmental pathogen exposure with a specific, self-limited response. In severe asthma and/or CRS, genome–environmental interactions lead to dysbiosis, aggravated inflammation, and disease. In healthy conditions, single or combined type 1, 2, and 3 immunological response pathways are invoked, generating cytokine, chemokine, innate cellular and T helper (Th) responses to eliminate viruses, helminths, and extracellular bacteria/fungi, correspondingly. Although the pathomechanisms are not fully known, the majority of severe airway diseases are related to type 2 high inflammation. Type 2 cytokines interleukins (IL) 4, 5, and 13, are orchestrated by innate lymphoid cell (ILC) and Th subsets leading to eosinophilia, immunoglobulin E (IgE) responses, and permanently impaired airway damage. Monoclonal antibodies can bind or block key parts of these inflammatory pathways, resulting in less inflammation and improved disease control.     

Coatings from micropatterned sulfobetaine polymer brushes as substrates for MC3T3-E1 cells

In the last decades, polymer brush coatings have proven to be excellent anti-fouling materials by preventing protein adhesion. When using this property to restrict cell growth laterally in cell culture, it is crucial to ensure that other cell functions remain unaffected. The present study therefore examines MC3T3-E1 cell growth and morphology on patterned PSBMA brush substrates and probes their proliferation potential at mRNA level. The osteoblastic cells display a more elongated morphology than cells on the control substrates, but show no sign of elevated levels of the apoptosis marker p53 or diminished levels of Ki-67 or H4, which serve as indicators of proliferation. Therefore, patterned polymer brushes do not seem to influence cells in their proliferation state and are suitable cell culture substrates. Nevertheless, the use of polymer brush surfaces in long-term cell culture was found to be limited by their instability in cell culture medium.