Archaeosomes for Oral Drug Delivery: From Continuous Microfluidics Production to Powdered Formulations.

Archaeosomes were manufactured from natural archaeal lipids by a microfluidics-assisted single-step production method utilizing a mixture of di- and tetraether lipids extracted from Sulfolobus acidocaldarius. The primary aim of this study was to investigate the exceptional stability of archaeosomes as potential carriers for oral drug delivery, with a focus on powdered formulations. The archaeosomes were negatively charged with a size of approximately 100 nm and a low polydispersity index. To assess their suitability for oral delivery, the archaeosomes were loaded with two model drugs: calcein, a fluorescent compound, and insulin, a peptide hormone. The archaeosomes demonstrated high stability in simulated intestinal fluids, with only 5% of the encapsulated compounds being released after 24 h, regardless of the presence of degrading enzymes or extremely acidic pH values such as those found in the stomach. In a co-culture cell model system mimicking the intestinal barrier, the archaeosomes showed strong adhesion to the cell membranes, facilitating a slow release of contents. The archaeosomes were loaded with insulin in a single-step procedure achieving an encapsulation efficiency of approximately 35%. These particles have been exposed to extreme manufacturing temperatures during freeze-drying and spray-drying processes, demonstrating remarkable resilience under these harsh conditions. The fabrication of stable dry powder formulations of archaeosomes represents a promising advancement toward the development of solid dosage forms for oral delivery of biological drugs.

A multi-step machine learning approach for accelerating QbD-based process development of protein spray drying.

This study proposes a new material-efficient multi-step machine learning (ML) approach for the development of a design space (DS) for spray drying proteins. Typically, a DS is developed by performing a design of experiments (DoE) with the spray dryer and the protein of interest, followed by deriving the DoE models via multi-variate regression. This approach was followed as a benchmark to the ML approach. The more complex the process and required accuracy of the final model is, the more experiments are necessary. However, most biologics are expensive and thus experiments should be kept to a minimum. Therefore, the suitability of using a surrogate material and ML for the development of a DS was investigated. To this end, a DoE was performed with the surrogate and the data used for training the ML approach. The ML and DoE model predictions were compared to measurements of three protein-based validation runs. The suitability of using lactose as surrogate was investigated and advantages of the proposed approach were demonstrated. Limitations were identified at protein concentrations >35 mg/ml and particle sizes of x50>6 µm. Within the investigated DS protein secondary structure was preserved, and most process settings, resulted in yields >75% and residual moisture <10 wt%.

Darwin's tales-A content analysis of how evolution is presented in children's books.

In science, certain theories led to a paradigm shift in human being's approach to explain nature, such as the theory of relativity, the quantum theory, and the theory of evolution. The latter explains the emergence of biodiversity on Earth and all living beings' relatedness, including humans. Accordingly, evolutionary theory is a central part of scientific literacy. However, scholars have demonstrated that misconceptions emerging in childhood hinder learners from grasping evolutionary processes. Implementing evolution in early science education could enhance scientific ideas as a basis for subsequent learning at school. Currently, children's literature that deals with evolution is increasing and may enable more children to encounter evolutionary theory before entering school. This explorative study aimed to analyze how children's books about evolution approach explaining this complex topic to young children in terms of covered contents, underlying concepts and use of language. We conducted (1) a text-based qualitative content analysis of 31 children's books in the categories of organismal context, evolutionary principles, and misconceptions, and (2) a computer-supported content analysis of 33 word labels concerning (a) scientific terms and (b) verbs expressing evolutionary change. Although evolution is a universal concept, children's books seem to promote specific contexts such as animal and human evolution. Even though the principle of selection requires an understanding of complex interactions between individuals and environmental factors, this principle was more frequent than the principles variation and inheritance. Phylogenetic history was covered more often than basic evolutionary processes, and evolutionary change was mainly mentioned at the species level over long periods. Besides, most books conveyed misconceptions such as transformationist, teleological or anthropomorphic reasoning. Consequently, books covering evolution may bias children's first ideas concerning this topic or introduce unscientific ideas. Based on our results, we propose implications for early evolution educators and education researchers.

In-vial printing and drying of biologics as a personalizable approach.

This study addressed the need for a flexible (personalizable) production of biologics, allowing their stabilization in the solid state and processing of small batch volumes. Therefore, inkjet printing into vials followed by a gentle vacuum drying step at ambient temperature was investigated by screening different formulations with a 22-full factorial design of experiments regarding printability. Human Serum Albumin (HSA) was used as a model protein in a wide range of concentrations (5 to 50 mg/ml), with (10 w/v%) and without the surfactant polysorbate 80 (PS80). PS80 was identified to positively affect the formulations by increasing the Ohnesorge number and stabilizing the printing process. The dispensed volumes with a target dose of 0.5 mg HSA were dried and analyzed concerning their residual moisture (RM) and protein aggregation. All investigated formulations showed an RM < 10 wt% and no significant induced protein aggregation as confirmed by Size Exclusion Chromatography (<2.5%) and Dynamic Light Scattering (Aggregation Index ≤ 2.5). Additionally, long-term printability and the available final dose after reconstitution were investigated for two optimized formulations. A promising formulation providing ∼93% of the targeted dose and a reconstitution time of 30 s was identified.

Comparing freeze drying and spray drying of interleukins using model protein CXCL8 and its variants.

Spray-dried products, such as synthetic peptides and hormones, have already been approved by the U.S. Food and Drug Agency and the European Medicines Agency, while spray-dried antibodies or interleukins, are not yet available on the market. Concerning the latter group, knowledge on whether and how spray-drying (SD) can be performed without adversely affecting their biological activity is lacking. Accordingly, this study aimed at establishing a SD process (Büchi B-90 spray dryer) using three Interleukin-8 based proteins (7-74 kDa) that were dispersed in phosphate buffered saline to maintain their stability. A Box-Behnken Design of Experiments was conducted to identify the appropriate process parameters taking into account the thermal stability of interleukin-8. In parallel, a FD process was developed. Both powders were stored for up to 12 weeks. Powder characterization included residual moisture evaluation and the mean particle size of the SD powder was investigated with Laser Diffraction Analysis. The hydrodynamic volume was measured via size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The secondary structure of the model proteins in the solid state was assessed with Fourier-transformation infrared spectroscopy for detecting the protein folding patterns and reconstituted with Circular Dichroism Spectroscopy. Finally, the binding affinity was studied with Surface Plasmon Resonance and Isothermal Fluorescence Titration, the protein stability with Chaotropic Unfolding, and the activity studies were carried out with the chemotaxis assay. The results showed that SD and FD powders with a residual moisture of less than 5 wt% were obtained. The interleukins showed no unfolding upon processing, neither in solid state nor reconstituted. Oligomerization was observed for FD, but not for SD interleukins. However, the unfolding, binding affinity and activity of all interleukins examined did not decrease in neither SD nor FD powders, even after 12 weeks of storage. Thus, it can be concluded that SD of interleukin formulations at outlet temperatures close to ambient temperature is a promising process for transferring them into a stable powder.

Conceptual Characterization of Threshold Concepts in Student Explanations of Evolution by Natural Selection and Effects of Item Context.

Evolutionary theory explains a wide range of biological phenomena. Proper understanding of evolutionary mechanisms such as natural selection is therefore an essential goal for biology education. Unfortunately, natural selection has time and again proven difficult to teach and learn, and students' resulting understanding is often characterized by misconceptions. Previous research has often focused on the importance of certain key concepts such as variation, differential survival, and change in population. However, so-called threshold concepts (randomness, probability, spatial scale, and temporal scales) have also been suggested to be important for understanding of natural selection, but there is currently limited knowledge about how students use these concepts. We sought to address this lack of knowledge by collecting responses to three different natural selection items from 247 university students from Sweden and Germany. Content analysis (deductive and inductive coding) and subsequent statistical analysis of their responses showed that they overall use some spatial scale indicators, such as individuals and populations, but less often randomness or probability in their explanations. However, frequencies of use of threshold concepts were affected by the item context (e.g., the biological taxa and trait gain or loss). The results suggest that the impact of threshold concepts, especially randomness and probability, on natural selection understanding should be further explored.

University Students' Conceptual Knowledge of Randomness and Probability in the Contexts of Evolution and Mathematics.

Students of all ages face severe conceptual difficulties regarding key aspects of evolution-the central, unifying, and overarching theme in biology. Aspects strongly related to abstract "threshold" concepts like randomness and probability appear to pose particular difficulties. A further problem is the lack of an appropriate instrument for assessing students' conceptual knowledge of randomness and probability in the context of evolution. To address this problem, we have developed two instruments, Randomness and Probability Test in the Context of Evolution (RaProEvo) and Randomness and Probability Test in the Context of Mathematics (RaProMath), that include both multiple-choice and free-response items. The instruments were administered to 140 university students in Germany, then the Rasch partial-credit model was applied to assess them. The results indicate that the instruments generate reliable and valid inferences about students' conceptual knowledge of randomness and probability in the two contexts (which are separable competencies). Furthermore, RaProEvo detected significant differences in knowledge of randomness and probability, as well as evolutionary theory, between biology majors and preservice biology teachers.