A functional high‐content miRNA screen identifies miR‐30 family to boost recombinant protein production in CHO cells

The steady improvement of mammalian cell factories for the production of biopharmaceuticals is a key challenge for the biotechnology community. Recently, small regulatory microRNAs (miRNAs) were identified as novel targets for optimizing Chinese hamster ovary (CHO) production cells as they do not add any translational burden to the cell while being capable of regulating entire physiological pathways. The aim of the present study was to elucidate miRNA function in a recombinant CHO‐SEAP cell line by means of a genome‐wide high‐content miRNA screen. This screen revealed that out of the 1, 139 miRNAs examined, 21% of the miRNAs enhanced cell‐specific SEAP productivity mainly resulting in elevated volumetric yields, while cell proliferation was accelerated by 5% of the miRNAs. Conversely, cell death was diminished by 13% (apoptosis) or 4% (necrosis) of all transfected miRNAs. Besides these large number of identified target miRNAs, the outcome of our studies suggest that the entire miR‐30 family substantially improves bioprocess performance of CHO cells. Stable miR‐30 over expressing cells outperformed parental cells by increasing SEAP productivity or maximum cell density of approximately twofold. Our results highlight the application of miRNAs as powerful tools for CHO cell engineering, identified the miR‐30 family as a critical component of cell proliferation, and support the notion that miRNAs are powerful determinants of cell viability.     

Cerebral Processing of Prosodic Emotional Signals: Evaluation of a Network Model Using rTMS

A great number of functional imaging studies contributed to developing a cerebral network model illustrating the processing of prosody in the brain. According to this model, the processing of prosodic emotional signals is divided into three main steps, each related to different brain areas. The present study sought to evaluate parts of the aforementioned model by using low-frequency repetitive transcranial magnetic stimulation (rTMS) over two important brain regions identified by the model: the superior temporal cortex (Experiment 1) and the inferior frontal cortex (Experiment 2). The aim of both experiments was to reduce cortical activity in the respective brain areas and evaluate whether these reductions lead to measurable behavioral effects during prosody processing. However, results obtained in this study revealed no rTMS effects on the acquired behavioral data. Possible explanations for these findings are discussed in the paper.     

High Selection Pressure Promotes Increase in Cumulative Adaptive Culture

The evolution of cumulative adaptive culture has received widespread interest in recent years, especially the factors promoting its occurrence. Current evolutionary models suggest that an increase in population size may lead to an increase in cultural complexity via a higher rate of cultural transmission and innovation. However, relatively little attention has been paid to the role of natural selection in the evolution of cultural complexity. Here we use an agent-based simulation model to demonstrate that high selection pressure in the form of resource pressure promotes the accumulation of adaptive culture in spite of small population sizes and high innovation costs. We argue that the interaction of demography and selection is important, and that neither can be considered in isolation. We predict that an increase in cultural complexity is most likely to occur under conditions of population pressure relative to resource availability. Our model may help to explain why culture change can occur without major environmental change. We suggest that understanding the interaction between shifting selective pressures and demography is essential for explaining the evolution of cultural complexity.     

Maternal Vitamin D Status in Preeclampsia: Seasonal Changes Are Not Influenced by Placental Gene Expression of Vitamin D Metabolizing Enzymes

Preeclampsia, a hypertensive disorder in pregnancy develops in 2–8% of pregnancies worldwide. Winter season and vitamin D deficiency have been associated with its onset.

Objective

To investigate the influence of season on maternal vitamin D status and placental vitamin D metabolism.

Methods

25-OH vitamin D and 1,25-(OH)₂ vitamin D were measured in maternal serum obtained during the winter or summer months from 63 pregnant women at delivery (43 healthy, 20 preeclampsia). In a subgroup, mRNA expression of CYP24A1 (24-hydroxylase), CYP27B1 (1α-hydroxylase) and VDR (vitamin D receptor) were quantified by real time PCR in placental samples of 14 women with normal pregnancies and 13 with preeclampsia.

Results

In patients with preeclampsia,25-OH vitamin D levels were lower, but differed significantly from controls only in summer (18.21±17.1 vs 49.2±29.2 ng/mL, P<0.001), whereas 1,25-(OH)₂ vitamin D levels were significantly lower only in winter (291±217 vs 612.3±455 pmol/mL, P<0.05). A two-factorial analysis of variance produced a statistically significant model (P<0.0001) with an effect of season (P<0.01) and preeclampsia (P = 0.01) on maternal 25-OH vitamin D levels, as well as a significant interaction between the two variables (P = 0.02). Placental gene expression of CYP24A1, CYP27B1, and VDR did not differ between groups or seasons. A negative correlation between placental gene expression of CYP24A1 and CYP27B1 was observed only in healthy controls (r = −0.81, P<0.0001).

Summary

Patients with preeclampsia displayed lower vitamin D serum levels in response to seasonal changes.The regulation of placental CYP24A1, but not of the VDR or CYP27B1 might be altered in preeclampsia.     

Population Size Predicts Lexical Diversity,but so Does the Mean Sea Level – Why It Is Important to Correctly Account for the Structure of Temporal Data

In order to demonstrate why it is important to correctly account for the (serial dependent) structure of temporal data, we document an apparently spectacular relationship between population size and lexical diversity: for five out of seven investigated languages, there is a strong relationship between population size and lexical diversity of the primary language in this country. We show that this relationship is the result of a misspecified model that does not consider the temporal aspect of the data by presenting a similar but nonsensical relationship between the global annual mean sea level and lexical diversity. Given the fact that in the recent past, several studies were published that present surprising links between different economic, cultural, political and (socio-)demographical variables on the one hand and cultural or linguistic characteristics on the other hand, but seem to suffer from exactly this problem, we explain the cause of the misspecification and show that it has profound consequences. We demonstrate how simple transformation of the time series can often solve problems of this type and argue that the evaluation of the plausibility of a relationship is important in this context. We hope that our paper will help both researchers and reviewers to understand why it is important to use special models for the analysis of data with a natural temporal ordering.     

ChipcheckII—Predicting Binding Curves for Multiple Analyte Strands on Small DNA Microarrays

Incomplete binding, saturation, and cross-hybridization between partially complementary strands complicate the parallel detection of nucleic acids via DNA microarrays. Treating the competing equilibria governing binding to microarrays requires computational tools. We have developed the web-based program ChipCheckII that calculates total hybridization matrices for target strands interacting with probes on small DNA microarrays. The program can be used to compute the extent of cross-hybridization and other phenomena affecting fidelity of detection based on sequences, quantities of strands, and hybridization conditions as inputs. Enthalpy and entropy of duplex formation are generated locally with UNAfold, including those for complexes that are partially matched. Simulated binding versus temperature curves for portions of a commercial genome chip demonstrate the extent to which cross-hybridization can complicate DNA detection. ChipCheckII is expected to aid nucleic acid chemists in developing high fidelity DNA microarrays.     

Regulation of circulating chromogranin B levels in heart failure

Background: Chromogranin B (CGB) regulates B-type natriuretic peptide (BNP) production. Circulating CGB levels are elevated in heart failure (HF) animal models and HF patients, but also increase in healthy individuals in response to physical activity. Therefore, CGB seems to integrate information from myocardial stress and systemic neuro-endocrine activation. Substantial gaps remain in our understanding of CGB regulation in HF.

Methods and results: We conducted a retrospective registry study including 372 patients. CGB and N-terminal pro-BNP (NT-proBNP) plasma levels were assessed in acute HF and chronic valvular HF patients and controls. CGB levels were significantly increased in acute HF and chronic valvular HF, but significantly higher in the latter. Patients in chronic valvular HF with severe mitral regurgitation (cHF-MR) showed significantly higher CGB levels than patients in chronic valvular HF with severe aortic stenosis. CGB levels progressively increased with worsening NYHA functional status and were moderately correlated to NT-proBNP, but independent of left ventricular (LV) ejection fraction (LVEF), LV mass, age and body weight. Finally, cHF-MR patients showed significant reductions of CGB levels after interventional mitral valve repair.

Conclusion: CGB is a promising emerging biomarker in HF patients with unique potential to integrate information from myocardial stress and neuro-endocrine activation.     

Thermo-Mechanical Properties of Soft Candy: Application of Time-Temperature Superposition to Mimic Response at High Deformation Rates

Soft candy primarily consists of an amorphous sugar matrix with highly adhesive and cohesive characteristics, which is generated by rapid cooling of a supersaturated sugar solution. The integration of fondant in soft candy leads to a partial recrystallization of the amorphous sugar so that the dispersed sugar crystals disrupt the continuous amorphous phase; this lowers cohesiveness, and creates a short texture that is responsible for a more brittle fracture. Final processing (e.g., cutting and packaging) is usually done at temperatures between 25 and 45 °C and short time scale. This study analyzed the effects of temperature and time scale on thermo-mechanical properties of different types of soft candy. The application of time-temperature superposition principle on small amplitude oscillatory shear experiments resulted in master curves that covered a frequency window up to 10⁵ rad/s, hence a time scale that is relevant in rapid processing. The respective shift factors depend on material properties, and a main factor of influence is the presence of a crystalline phase. Simple penetration and tensile tests give additional information on material behavior, especially with respect to effects of temperature on adhesiveness and cohesiveness. The results of the study provide support for further optimizing soft candy formulations to ensure rapid and undisturbed processing.     

Mechanisms preventing a decrease in phytoplankton biomass after phosphorus reductions in a German drinking water reservoir—results from more than 50 years of observation

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Inhibition of Photosystem II by UV-B Radiation and the Conditions for Recovery in the Liverwort Conocephalum conicum Dum.

Inhibition of photosynthesis by UV-B was investigated in the thalloid liverwort Conocephalum conicum Dum. UV-B irradiance was adjusted to a strength producing 50% inhibition of the rate of photosynthesis during 10 min of irradiation. A linear relationship of the fluorescence terms F_v/F_m of photosystem (PS) II and J_P was observed following a UV-B irradiation. This suggested that PS II was a major site of UV-B-induced damage of photosynthesis. The apparent inhibition of F_v/F_m was much smaller when electron flow to the secondary PS II acceptor Q_B was inhibited by DCMU or when F_v/F_m was measured at 77 K. Apparently, the major target of UV-B effects was electron donation to the PS II reaction center, rather than electron transfer reactions at the PS II acceptor side. The time required for repair of PS II from UV-B-induced damage was light-dependent and minimal at a flux density of 5 μE m^?2 s^?1. Low temperatures and the presence of streptomycin inhibited the repair processes of PS II, indicating that protein synthesis may be involved in the recovery of PS II. The data indicate that UV-B irradiation on bright and cool winter days may be most harmful for photosynthesis of C. conicum. A repeated irradiation of the thalli with UV-B induced tolerance of photosynthesis which was related to an accumulation of pigments with a maximum of absorption around 315 nm.