Glucose-limited high cell density cultivations from small to pilot plant scale using an enzyme-controlled glucose delivery system

The enzyme controlled substrate delivery cultivation technology EnBase(?) Flo allows a fed-batch-like growth in batch cultures. It has been previously shown that this technology can be applied in small cultivation vessels such as micro- and deep well plates and also shake flasks. In these scales high cell densities and improved protein production for Escherichia coli cultures were demonstrated. This current study aims to evaluate the scalability of the controlled glucose release technique to pilot scale bioreactors. Throughout all scales, that is, deep well plates, 3 L bioreactor and 150 L bioreactor cultivations, the growth was very similar and the model protein, a recombinant alcohol dehydrogenase (ADH) was produced with a high yield in soluble form. Moreover, EnBase Flo also was successfully used as a controlled starter culture in high cell density fed-batch cultivations with external glucose feeding. Here the external feeding pump was started after overnight cultivation with EnBase Flo. Final optical densities in these cultivations reached 120 (corresponding to about 40 g L(-1) dry cell weight) and a high expression level of ADH was obtained. The EnBase cultivation technology ensures a controlled initial cultivation under fed-batch mode without the need for a feeding pump. Because of the linear cell growth under glucose limitation it provides optimal and robust starting conditions for traditional external feed-based processes.     

Zeitgebers and their association with rest-activity patterns

Zeitgebers such as light, eating and physical activity provide input to the circadian clock. Chronic circadian misalignment is associated with significant adverse health effects. An improved understanding of the impact of the timing of zeitgebers on the stability of 24-hour rest-activity rhythm in free-living settings may identify behavioural and environmental intervention targets. A total of 133 healthy adults, aged 21–60 years, wore a wrist actigraph for 7 consecutive days. We applied a non-parametric analysis to activity counts to derive rest-activity patterns. We administered a questionnaire through a smartphone app to collect self-reported timing of light exposure, eating episodes and physical activity. To assess the relationship between timing exposures (first and last exposure to outdoor light, first exposure to indoor light, last eating episode, first eating episode, morning physical activity proportion, evening physical activity proportion) and rest-activity or sleep outcomes (bedtimes, total sleep time, inter-daily stability, intra-daily variability, L5 and M10 midpoint), we first calculated Spearman correlations, using the false discovery rate method to control for multiple comparisons. From those significant associations, we then fit regression models adjusting for age, sex, race, household income, education level, study site, body mass index, as well as physical activity. Finally, we tested for interaction between chronotype and each timing-related exposure and stratified the analysis by morning type. All zeitgebers, except for evening physical activity proportion, were correlated with at least four of the seven sleep and rest-activity outcomes. In adjusted analysis, later timing of first (after 6:30 to 7:45 AM versus earlier) and last exposure to indoor light (after 11:00 PM versus earlier) and first (after 7:45–9:45 AM versus earlier) and last eating episode (after 8:00–09:00 PM versus earlier) were associated with a shift of 0.60–1.39 hours to later bedtimes, M10 and L5 midpoints (i.e. timing of peak activities or inactivities). Later timing of first exposure to outdoor light (after 09:30 AM versus earlier) was also associated with 0.51 (95% CI: 0.19 to 0.83) hours longer total sleep time. Higher morning physical activity proportion (> 33%) was associated with 0.95 (95% CI: ?1.38 to ?0.53) hours earlier in-bed time and 0.69 (95% CI: ?1.14 to ?0.24) hours earlier out-of-bed time, 0.92 (95% CI: ?1.41 to ?0.42) hours earlier M10 and 0.96 (95% CI: ?1.42 to ?0.49) min earlier L5 midpoint. The results did not change substantially with further adjustment for total activity. There was a significant interaction between morning chronotype and first eating episode with rest-activity patterns (p < 0.05), with first eating episode associating with timing of activities only in non-morning type adults. Timing of zeitgebers was associated with sleep and rest-activity patterns, including bedtimes, L5 and M10 midpoint. Future research should evaluate the impact of manipulating zeitgebers on both circadian rhythms and health outcomes.     

Biology student teachers’ dialogic talk in inquiry-based instruction

ABSTRACT

Inquiry-based learning has generally accepted by scholars as a most effective teaching approach in biology education. The talk during inquiry-based teaching needs to be practiced. There is less evidence how student teachers talk with students during their inquiry-based biology instruction. This knowledge is needed in supporting student teachers to develop their teachership in biology education. In this qualitative case study, the dialogic talk of biology student teachers (N = 6) was studied in the context of inquiry-based lessons in lower secondary school. The student teachers’ lessons were video and audio recorded and the data was analyzed using content analysis. The student teachers used dialogic talk in their inquiry-based instruction only occasionally, mainly in the examination and the conclusion stages. During the introduction stage, dialogic talk was less used and it was mainly explaining and instructing the content. In the examination stage, student teachers also guided students and stated facts. During the conclusion stage, student teachers mainly explained and also evaluated students’ statements. The lesson’s topics and methods used in inquiry-based learning may enable the dialogic talk of student teacher to some extent. However, teacher education should focus more on scaffolding student teachers’ talk with their students in all kinds of inquiry approaches.     

The chemical character and burial of phosphorus in shallow coastal sediments in the northeastern Baltic Sea

The chemical composition and vertical distribution of sediment phosphorus (P) in shallow coastal sediments of the northeastern Baltic Sea (BS) were characterized by sequential extraction. Different P forms were related to chemical and physical properties of the sediments and the chemistry of pore water and near-bottom water. Sediment P composition varied among the sampling sites located in the Archipelago Sea (AS) and along the northern coast of the Gulf of Finland (GoF): the organic rich sites were high in organic P (OP), while apatite-P dominated in the area affected by sediment transportation. Although the near-bottom water was oxic, the sediments released P. Release of P was most pronounced at the site with high sediment OP and reduced conditions in the sediment-water interface, indicating that P had its origins in organic sources as well as in reducible iron (Fe) oxyhydroxides. The results suggest that even though these coastal areas are shallow enough to lack salinity stratification typical for the brackish BS, they are vulnerable to seasonal oxygen (O₂) depletion and P release because of their patchy bottom topography, which restricts mixing of water. Furthermore, coastal basins accumulate organic matter (OM) and OP, degradation of which further diminishes O₂ and creates the potential for P release from the sediment. In these conditions, an abundance of labile OP may cause marked efflux of P from sediment reserves in the long-term.     

Liver-Fat Accumulation and Insulin Resistance in Obese Women with Previous Gestational Diabetes

Objective: We determined whether fat accumulation in the liver is associated with features of insulin resistance independent of obesity. Research Methods and Procedures: We recruited 27 obese nondiabetic women in whom liver fat (LFAT) content was determined by proton spectroscopy, intra-abdominal and subcutaneous fat by magnetic resonance imaging, and insulin sensitivity by the euglycemic insulin clamp technique. The women were divided based on their median LFAT content (5%) to groups with low (3.2 ± 0.3%) and high (9.8 ± 1.5%) liver fat. The groups were almost identical with respect to age (36 ± 1 vs. 38 ± 1 years in low vs. high-LFAT), body mass index (32.2 ± 0.6 vs. 32.8 ± 0.5 kg/m²), waist-to-hip ratio, intra-abdominal, subcutaneous, and total fat content. Results: Women with high LFAT had features of insulin resistance including higher fasting serum triglyceride (1.93 ± 0.21 vs. 1.11 ± 0.09 mM, p < 0.01) and insulin (14 ± 3 vs. 10 ± 1 mU/L, p < 0.05) concentrations than women with low LFAT. The group with high LFAT also had higher 24-hour blood pressures, and lower whole-body insulin sensitivity compared with the low-LFAT group. Discussion: In obese women with previous gestational diabetes, LFAT, rather than any measure of body composition, is associated with features of insulin resistance.