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Background: Mesocosm experiments have become increasingly popular in climate change research as they bridge the gap between small-scale, less realistic, microcosm experiments, and large-scale, more complex, natural systems. Characteristics of aquatic mesocosm designs (e.g., mesocosm volume, study duration, and replication) vary widely, potentially affecting the magnitude and direction of effect sizes measured in experiments. In this global systematic review we aim to identify the type, direction and strength of climate warming effects on aquatic species, communities and ecosystems in mesocosm experiments. Furthermore, we will investigate the context-dependency of the observed effects on several a priori determined effect moderators (ecological and methodological). Our conclusions will provide recommendations for aquatic scientists designing mesocosm experiments, as well as guidelines for interpretation of experimental results by scientists, policy-makers and the general public. Methods: We will conduct a systematic search using multiple online databases to gather evidence from the scientific literature on the effects of warming experimentally tested in aquatic mesocosms. Data from relevant studies will be extracted and used in a random effects meta-analysis to estimate the overall effect sizes of warming experiments on species performance, biodiversity and ecosystem functions. Experimental characteristics (e.g., mesocosm size and shape, replication-level, experimental duration and design, biogeographic region, community type, crossed manipulation) will be further analysed using subgroup analyses.

The demand for sustainable textiles has increased research on bio-based biopolymers. Fiber spinning from starch biopolymer and its blends was widely attempted using electro-spinning; however, it is less attempted using melt spinning which is an industrially feasible method. In this study, native tapioca starch is converted into thermoplastic starch (TPS), blended with polylactic acid (PLA), and is attempted for melt spinnability in its neat and blended form TPS/PLA (70/30) with the intention of checking the possibility of melt spinning. The results from characterization of the prepared biopolymers show that thermal analysis of neat TPS does not reveal clear thermal transitions, glass transition (T g), and melting (T m), in the second heating curve, rather it influenced crystallization behavior of PLA as seen from differential scanning calorimetry result and degradation temperature (T d) was found to be in the range of 296–352°C from thermogravimetric analysis showing that addition of PLA improved thermal stability of TPS. Morphology analysis with AFM images revealed the presence of granular starch in neat TPS and phase separation in TPS/PLA blends with finer phase distribution in the presence of additives. Fourier transform infrared spectroscopy result shows the interaction between starch, glycerol, and PLA showing the effect of thermoplasticization of starch which brings rupture or weakening of the strong glycosidic bonds in between starch molecules and interaction between TPS and PLA as it can be seen from peak shift and peak intensity. The melt spinning trials show the possibility of melt spinning TPS-based biopolymers into fibers even with higher content of TPS as well as neat TPS giving an insight and motivation for more research engagement. The melt-spun fibers were found to have a diameter in the range of 160.0–117.0 μm. However, additional experiments and investigations are required to improve the mechanical and other properties of the fibers.

Plant responses to environmental heterogeneity depend on life-history traits, which could relate to phenotypical and genetic characteristics. To elucidate this relationship, we examined the variation in population genetics and functional traits of short- and long-lived Artemisia species that are co-occurring in the steppes of Mongolia. Mongolian steppes represent stressful and water-limited habitats, demanding phenotypic modifications in the short term and/or genetic adaptation in the long term. However, detailed knowledge is missing about both plant phenotypic and genetic differentiation, and their interrelationships in temperate grasslands. Here, we investigated 21 populations of the widely distributed subshrub Artemisia frigida and the herbaceous biennial Artemisia scoparia . Genetic variation was assessed with newly developed simple sequence repeats (SSRs) markers. Functional trait data were collected from each individual, and data on environmental variables was collected for each population. We detected significantly higher genetic diversity in the biennial species ( H E  = 0.86) compared with the perennial ( H E  = 0.79). For both species, the largest share of genetic variation was partitioned within populations (96%). Population genetic structure in the biennial A. scoparia was weak, while the perennial A. frigida showed some spatial genetic structure, which was impacted by geographical factors, soil nutrients, and precipitation amount. Morphology-related functional traits (i.e., plant height) were predominantly associated with environmental variables rather than with genetic variation, whereas physiology-related trait (i.e., specific leaf area [SLA]) was partly genetically determined., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Khurelpurev Oyundelger reports financial support was provided by TU Dresden. Karsten Wesche reports travel was provided by German Federal Ministry of Education and Research. Christiane Ritz reports travel was provided by German Academic Exchange Service., (© 2024 The Author(s). Ecology and Evolution published by John Wiley & Sons Ltd.)

This study focused on the production of protein isolates from mantis shrimp (MS). The pH-shift method was investigated to understand its impact on the protein yield, quality, and properties of the produced isolates. The first step was determining how the pH affected the protein solubility profile, zeta potential, and brown discoloration. The pH-shift process was then established based on the maximum and minimum protein solubilization. The solubilization pH had a significant impact on the mass yield and color of the produced protein, with a pH of 1.0 producing the maximum mass in the acidic region, whereas a maximum was found at a pH of 12.0 in the alkaline region ( p < 0.05). Both approaches yielded mantis shrimp protein isolates (MPIs) with precipitation at a pH of 4.0 and a mass yield of around 25% (dw). The TCA-soluble peptide and TBARS levels were significantly lower in the MPI samples compared to MS raw material ( p < 0.05). The MPIs maintained essential amino acid index (EAAI) values greater than 90%, indicating a high protein quality, and the pH-shift procedure had no negative impact on the protein quality, as indicated by comparable EAAI values between the mantis shrimp protein isolate extract acid (MPI-Ac), mantis shrimp protein isolate extract alkaline (MPI-Al), and MS raw material. Overall, the pH-shift approach effectively produced protein isolates with favorable quality and nutritional attributes.

We have recently shown that cancer cells of various origins take up extracellular citrate through the plasma membrane citrate carrier (pmCiC), a specific plasma membrane citrate transporter. Extracellular citrate is required to support cancer cell metabolism, in particular fatty acid synthesis, mitochondrial activity, protein synthesis and histone acetylation. In addition, cancer cells tend to acquire a metastatic phenotype in the presence of extracellular citrate. Our recent study also showed that cancer-associated stromal cells synthesise and release citrate and that this process is controlled by cancer cells. In the present study, we evaluated the expression of pmCiC, fibroblast activation protein-α (FAP) and the angiogenesis marker cluster of differentiation 31 (CD31) in human cancer tissues of different origins. In the cohort studied, we found no correlation between disease stage and the expression of FAP or CD31. However, we have identified a clear correlation between pmCiC expression in cancer cells and cancer-associated stroma with tumour stage. It can be concluded that pmCiC is increased in cancer cells and in cancer-supporting cells in the tumour microenvironment at the later stages of cancer development, particularly at the metastatic sites. Therefore, pmCiC expression has the potential to serve as a prognostic marker, although further studies are needed., Competing Interests: MM is an inventor on the Patent Application no. EP15767532.3 and US15/514,255 (status patent pending) “The plasma membrane citrate transporter for use in the diagnosis and treatment of cancer” owned by the University Hospital Regensburg. GZ is an employee and shareholder of Eternygen GmbH. MM is a shareholder of MetaSMIte, Sp. z o.o., Krakow, Poland. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The authors declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Schwertner, Dahdal, Jagla, Grossmann, Drexler, Krahn, Evert, Berneburg, Haferkamp, Ziegler, Parkinson, Zahn, Mycielska and Gaumann.)

Background: Neuroblastoma is a heterogeneous disease with adrenergic (ADRN)- and therapy resistant mesenchymal (MES)-like cells driven by distinct transcription factor networks. Here, we investigate the expression of immunotherapeutic targets in each neuroblastoma subtype and propose pan-neuroblastoma and cell state specific targetable cell-surface proteins., Methods: We characterized cell lines, patient-derived xenografts, and patient samples as ADRN-dominant or MES- dominant to define subtype-specific and pan-neuroblastoma gene sets. Targets were validated with ChIP- sequencing, immunoblotting, and flow cytometry in neuroblastoma cell lines and isogenic ADRN-to-MES transition cell line models. Finally, we evaluated the activity of MES-specific agents in vivo and in vitro ., Results: Most immunotherapeutic targets being developed for neuroblastoma showed significantly higher expression in the ADRN subtype with limited expression in MES-like tumor cells. In contrast, CD276 (B7-H3) and L1CAM maintained expression across both ADRN and MES states. We identified several receptor tyrosine kinases (RTKs) enriched in MES-dominant samples and showed that AXL targeting with ADCT-601 was potently cytotoxic in MES-dominant cell lines and showed specific anti-tumor activity in a MES cell line-derived xenograft., Conclusions: Immunotherapeutic strategies for neuroblastoma must address the potential of epigenetic downregulation of antigen density as a mechanism for immune evasion. We identified several RTKs as candidate MES-specific immunotherapeutic target proteins for the elimination of therapy-resistant cells. We hypothesize that the phenomena of immune escape will be less likely when targeting pan-neuroblastoma cell surface proteins such as B7-H3 and L1CAM, and/or dual targeting strategies that consider both the ADRN- and MES-cell states., Key Points: Cellular plasticity influences the abundance of immunotherapeutic targets.Subtype-specific targets may be susceptible to epigenetically-mediated downregulation.Immunotherapeutic targets in development, B7-H3 and L1CAM, show "pan-subtype" expression., Importance of Study: Neuroblastoma is a lethal childhood malignancy that shows cellular plasticity in response to anti-cancer therapies. Several plasma membrane proteins are being developed as immunotherapeutic targets in this disease. Here we define which cell surface proteins are susceptible to epigenetically regulated downregulation during an adrenergic to mesenchymal cell state switch and propose immunotherapeutic strategies to anticipate and circumvent acquired immunotherapeutic resistance.

The global food system is shifting towards cellular agriculture, a second domestication marked by cultivating microorganisms and tissues for sustainable food production. This involves tissue engineering, precision fermentation, and microbial biomass fermentation to establish food value chains independent of traditional agriculture. However, these techniques rely on growth media sourced from agricultural, chemical (fossil fuels), and mining supply chains, raising concerns about land use competition, emissions, and resource depletion. Fermentable sugars, nitrogen, and phosphates are key ingredients derived from starch crops, energy-intensive fossil fuel based processes, and finite phosphorus resources, respectively. This review explores sustainable alternatives to reduce land use and emissions associated with cellular agriculture media ingredients. Sustainable alternatives to first generation sugars (lignocellulosic substrates, sidestreams, and gaseous feedstocks), sustainable nitrogen sources (sidestreams, green ammonia, biological nitrogen fixation), and efficient use of phosphates are reviewed. Especially cellulosic sugars, gaseous chemoautotrophic feedstocks, green ammonia, and phosphate recycling are the most promising technologies but economic constraints hinder large-scale adoption, necessitating more efficient processes and cost reduction. Collaborative efforts are vital for a biotechnological future grounded in sustainable feedstocks, mitigating competition with agricultural land and emissions., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Internet-based cognitive behavioral therapy (iCBT) offers a scalable, cost-effective, accessible, and low-threshold form of psychotherapy. Recent advancements explored the use of conversational agents such as chatbots and voice assistants to enhance the delivery of iCBT. These agents can deliver iCBT-based exercises, recognize and track emotional states, assess therapy progress, convey empathy, and potentially predict long-term therapy outcome. However, existing systems predominantly utilize categorical approaches for emotional modeling, which can oversimplify the complexity of human emotional states. To address this, we developed a transformer-based model for dimensional text-based emotion recognition, fine-tuned with a novel, comprehensive dimensional emotion dataset comprising 75,503 samples. This model significantly outperforms existing state-of-the-art models in detecting the dimensions of valence, arousal, and dominance, achieving a Pearson correlation coefficient of r  = 0.90, r  = 0.77, and r  = 0.64, respectively. Furthermore, a feasibility study involving 20 participants confirmed the model's technical effectiveness and its usability, acceptance, and empathic understanding in a conversational agent-based iCBT setting, marking a substantial improvement in personalized and effective therapy experiences., Competing Interests: The authors declare there are no competing interests., (©2024 Striegl et al.)

Besides teachers' professional knowledge, their self-efficacy is a crucial aspect in promoting students' scientific reasoning (SR). However, because no measurement instrument has yet been published that specifically refers to self-efficacy beliefs regarding the task of teaching SR, we adapted the Science Teaching Efficacy Belief Instrument (STEBI) accordingly, resulting in the Teaching Scientific Reasoning Efficacy Beliefs Instrument (TSR-EBI). While the conceptual framework of the TSR-EBI is comparable to that of the STEBI in general terms, it goes beyond it in terms of specificity, acknowledging the fact that teaching SR requires very specific knowledge and skills that are not necessarily needed to the same extent for promoting other competencies in science education. To evaluate the TSR-EBI's psychometric quality, we conducted two rounds of validation. Both samples ( N 1 = 114; N 2 = 74) consisted of pre-service teachers enrolled in university master's programs in Germany. The collected data were analyzed by applying Rasch analysis and known-group comparisons. In the course of an analysis of the TSR-EBI's internal structure, we found a 3-category scale to be superior to a 5-category structure. The person and item reliability of the scale proved to be satisfactory. Furthermore, during the second round of validation, it became clear that the results previously found for the 3-category scale were generally replicable across a new (but comparable) sample, which clearly supports the TSR-EBI's psychometric quality. Moreover, in terms of test-criterion relationships, the scale was also able to discriminate between groups that are assumed to have different levels of self-efficacy regarding teaching SR. Nonetheless, some findings also suggest that the scale might benefit from having the selection of individual items reconsidered (despite acceptable item fit statistics). On balance, however, we believe that the TSR-EBI has the potential to provide valuable insights in future studies regarding factors that influence teachers' self-efficacy, such as their professional experiences, prior training, or perceived barriers to effective teaching., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Welter, Dawborn-Gundlach, Großmann and Krell.)

The development of novel, sustainable, and robust food production technologies represents one of the major pillars to address the most significant challenges humanity is going to face on earth in the upcoming decades - climate change, population growth, and resource depletion. The implementation of microfoods, i.e., foods formulated with ingredients from microbial cultivation, into the food supply chain has a huge potential to contribute towards energy-efficient and nutritious food manufacturing and represents a means to sustainably feed a growing world population. This review recapitulates and assesses the current state in the establishment and usage of gas fermenting bacteria as an innovative feedstock for protein production. In particular, we focus on the most promising representatives of this taxon: the hydrogen-oxidizing bacteria (hydrogenotrophs) and the methane-oxidizing bacteria (methanotrophs). These unicellular microorganisms can aerobically metabolize gaseous hydrogen and methane, respectively, to provide the required energy for building up cell material. A protein yield over 70% in the dry matter cell mass can be reached with no need for arable land and organic substrates making it a promising alternative to plant- and animal-based protein sources. We illuminate the holistic approach to incorporate protein extracts obtained from the cultivation of gas fermenting bacteria into microfoods. Herein, the fundamental properties of the bacteria, cultivation methods, downstream processing, and potential food applications are discussed. Moreover, this review covers existing and future challenges as well as sustainability aspects associated with the production of microbial protein through gas fermentation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

The objective of this research was to improve the protein extraction processes of Rastrelliger kanagurta (Indian mackerel) to generate protein isolate with enhanced bio-physico-chemical properties and gel-forming ability. To achieve this, two novel approaches were designed that utilized an additional alkaline separation step and were compared to a conventional process: acid solubilization → alkaline solubilization → pI and acid solubilization → pI → alkaline solubilization. The novel extraction designs resulted in a lower lipid content, lipid oxidation, and TCA-soluble peptides, as well as improving the color and sensory features of the refined proteins, which corresponded to the lowest total heme pigments ( p < 0.05). Furthermore, the protein isolate recovered with the modified processes showed significant changes in biochemical properties (decreases in Ca 2+ -ATPase activity/reactive sulfhydryl content and an increase in surface hydrophobicity) and dynamic rheological behavior. As a result, by altering the extraction procedure it was possible to obtain improved gel characteristics such as gel strength, color, expelled moisture, and improved gel microstructure. Moreover, this study demonstrated that the gel network was partly stabilized by disulfide bonds, according to SDS-PAGE. Overall, this study demonstrates that by optimizing protein extraction procedures a considerable improvement in quality can be achieved and that an additional alkaline extraction after isoelectric point precipitation results in the optimized gel-forming ability of mackerel proteins.

Pedagogical content knowledge (PCK) is one core dimension of teachers' professional knowledge and comprises knowledge about conceptual ideas of learners and appropriate instructions. However, several challenges regarding the assessment of PCK are discussed in the literature: For example, PCK is a topic-specific construct and contains differentiable subdomains, which must be considered during test development. In addition, the choice of test type needs to be considered. While open-ended instruments can capture a broader range of cognitions, they often require a high level of interpretation; in contrast, multiple-choice instruments have advantages in terms of objectivity and test economy. Some challenges of assessing PCK are particularly related to multiple-choice instruments, such as an insufficient focus on specific components or the accidental assessment of teachers' beliefs instead of PCK. To better understand and explain these challenges in developing multiple-choice PCK instruments, we exemparly used an instrument to assess PCK about scientific reasoning and considered the assumptions of the expert-novice paradigm to analyze differential response behavior between n  = 10 researchers in the field of biology education (experts) and n  = 10 undergraduate pre-service biology teachers (novices). As expected, experts scored significantly higher than novices. At the same time, experts answered the items more consistently than novices, i.e., showed less variance. However, the difference found was statistically insignificant. Regarding the explanations for choosing a response option, experts more often correctly identified the quintessence of the items, which means that they more often understand the items as intended and argued based on their PCK. On the other hand, novices focused more on surface characteristics, i.e., they argued rather with surface knowledge like intuition or personal experience, than choosing the response option based on their PCK. These crucial differences in how experts and novices understand the items of the used PCK instrument and how they respond based on their understanding affect different test characteristics. In conclusion, we recommend ensuring that instruments address only a few, specific PCK aspects, considering the target group of a test, and take into account that target groups with larger variability among their responses require a higher number of items to achieve satisfactory discrimination and reliability indices., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Lieberei, Welter, Großmann and Krell.)

Salted eggs are normally produced by treating fresh duck eggs with a high salt concentration in order to acquire distinctive features and excellent preservation capabilities as a result of a series of physicochemical changes. This method, however, induces a high salt content in the product. The goal of this research was to create a new way of producing mildly salted duck eggs using ozonized brine salting. The brine was made by dissolving NaCl (26% w / v ) in water or ozonized water at a concentration of 50 ng ozone/mL (ozonized brine). Compared to brine, ozonized brine resulted in salted eggs with reduced ultimate salt levels in both albumen and yolk ( p < 0.05). The Haugh unit of the salted eggs generated by ozonized brine was similar to that of the brine-made salted egg group ( p > 0.05), but the salted egg produced by ozonized brine matured and solidified faster because the yolk index (0.62) was higher than that of the brine (0.55) ( p < 0.05). The final pH of salted eggs generated with brine and ozonized brine was not different ( p > 0.05). Regardless of the salting method, both salted eggs contained low TVB-N content (<10 mg/100 g). Ozonized brine increased the protein carbonyl content in salted albumen, which may be related to albumen protein aggregation and served as a salt diffusion barrier. However, after boiling the salted egg, the protein carbonyl level was comparable to that of fresh albumen. The TBARS levels of boiled salted albumen prepared with brine and ozonized brine were comparable ( p > 0.05), and the value was extremely low (~0.1 mg MDA equivalent/kg). The TBARS value of the salted yolk prepared with brine was higher than that of the salted yolk prepared with ozonized brine ( p < 0.05), and both salted yolks showed increased TBARS values after cooking ( p < 0.05). The albumen and yolk components appeared to be altered similarly by both brine and ozonized brine, according to the FTIR spectra. Furthermore, the appearance and color of the yolk and albumen in salted eggs prepared with brine and ozonized brine were comparable. Boiled salted albumen produced with ozonized brine had a denser structure with fewer voids. This could be attributed to the final salted egg's lower salt content and lower salt diffusion rate, which were likely caused by protein oxidation and, as a result, aggregation when ozonized brine was used.

A growing number of health-conscious consumers are looking for animal protein alternatives with similar texture, appearance, and flavor. However, research and development still needs to find alternative non-meat materials. The aim of this study was to develop a mushroom-based minced meat substitute (MMMS) from edible Pleurotus sajor-caju (PSC) mushrooms and optimize the concentration of chickpea flour (CF), beetroot extract, and canola oil. CF was used to improve the textural properties of the MMMS by mixing it with PSC mushrooms in ratios of 0:50, 12.5:37.5, 25:25, 37.5:12.5, and 50:0. Textural and sensory attributes suggest that PSC mushrooms to CF in a ratio of 37.5:12.5 had better textural properties, showing hardness of 2610 N and higher consumer acceptability with protein content up to 47%. Sensory analysis suggests that 5% ( w/w) canola oil showed the most acceptable consumer acceptability compared to other concentrations. Color parameters indicate that 0.2% beetroot extract shows higher whiteness, less redness, and higher yellowness for both fresh and cooked MMMS. This research suggests that MMMS containing PSC, CF, canola oil, and beetroot extract could be a suitable alternative and sustainable food product which may lead to higher consumer adoption as a meat substitute.

In recent years, brewer's spent grain (BSG) has gained attention as a plant-based protein source because it occurs in large quantities as a by-product of beer brewing. BSG can contribute to future food requirements and support the development of a circular economy. In light of the dynamic developments in this area, this review aims to understand the proteins present in BSG, and the effect of extraction techniques and conditions on the composition, physicochemical, and techno-functional properties of the obtained protein extracts. The water-insoluble hordeins and glutelins form the major protein fractions in BSG. Depending on the beer brewing process, the extraction technique, and conditions, the BSG protein isolates predominantly contain B, C, and ϒ hordeins, and exhibit a broad molecular weight distribution ranging between <5 kDa and >250 kDa. While the BSG isolates obtained through chemical extraction methods seem promising to obtain gelled food products, physical and enzymatic modifications of BSG proteins through ultrasound and proteolytic hydrolysis offer an effective way to produce soluble and functional protein isolates with good emulsifying and foaming capabilities. Specifically tailored protein extracts to suit different applications can thus be obtained from BSG, highlighting that it is a highly valuable protein source.

Fibers and yarns are part of everyday life. So far, fibers that are also used pharmaceutically have mainly been produced by electrospinning. The common use of spinning oils and the excipients they contain, in connection with production by melt extrusion, poses a regulatory challenge for pharmaceutically usable fibers. In this publication, a newly developed small-scale direct-spinning melt extrusion system is described, and the pharmaceutically useful polyvinyl filaments produced with it are characterized. The major parts of the system were newly developed or extensively modified and manufactured cost-effectively within a short time using rapid prototyping (3D printing) from various materials. For example, a stainless-steel spinneret was developed in a splice design for a table-top melt extrusion system that can be used in the pharmaceutical industry. The direct processing of the extruded fibers was made possible by a spinning system developed called Spinning-Rosi, which operates continuously and directly in the extrusion process and eliminates the need for spinning oils. In order to prevent instabilities in the product, further modifications were also made to the process, such as a the moisture encapsulation of the melt extrusion line at certain points, which resulted in a bubble-free extrudate with high tensile strength, even in a melt extrusion line without built-in venting.

Hypothesis: The emulsifying ability of starch is influenced by its molecular weight. Reducing the molecular weight of starch is expected to influence interfacial adsorption and membrane elasticities, thereby affecting its emulsifying ability through Pickering effects. Hence, it should be possible to tailor the emulsifying ability of starch by adjusting its molecular weight., Experiments: Waxy corn starch (CS) and rice starch (RS) were hydrolyzed with pullulanase to obtain high (HM) and low molecular weight (LM) fractions. After the molecular weight was determined by size exclusion chromatography, the fractions were used to prepare model oil-in-water emulsions. The stability, microscopy, and particle size of the emulsions were characterized, and the underlying emulsification mechanism was subsequently studied through dynamic laser scattering, surface tension analysis, interfacial rheology, and Pearson's correlation calculations., Findings: In the molecular weight range obtained in this study, the smaller the molecular weight of starch, the stronger its emulsifying ability. The decrease in molecular weight resulted in considerable different adsorption and interfacial elasticities with smaller fractions occupying less area on the interface and forming interfaces with higher elasticities, resulting in higher stabilities through Pickering effects. Results thus suggest that the emulsifying ability of starch may be tailored by adjusting its molecular weight., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The combination of polymerization-induced self-assembly (PISA) and reversible-addition fragmentation chain transfer (RAFT) emulsion polymerization offers a powerful technique to synthesize diblock copolymers and polymeric nanoparticles in a controlled manner. The RAFT emulsion diblock copolymerization of styrene and methacrylic acid (MAA) by using a trithiocarbonate as surfactant and RAFT agent was investigated. The Z-group of the RAFT agent was modified with a propyl-, butyl- and dodecyl- sidechain, increasing the hydrophobicity of the RAFT agent to offer well-controlled polymerization of poly(methacrylic acid)- block -polystyrene (PMAA- b -PS) diblock copolymers at high solid contents between 30-50 wt% in water. The kinetic data of the PMAA homopolymerization with the three different RAFT agents for various solvents was investigated as well as the RAFT emulsion polymerization of the diblock copolymers in pure water. While the polymerization of PMAA- b -PS with a propyl terminus as a Z-group suffered from slow polymerization rates at solid contents above 30 wt%, the polymerization with a dodecyl sidechain as a Z-group led to full conversion within 2 h, narrow molar mass distributions and all that at a remarkable solid content of up to 50 wt%.

An in vitro dissolution model, the so-called EyeFlowCell (EFC), was developed to test intravitreal dosage forms, simulating parameters such as the gel-like consistency of the vitreous body. The developed model consists of a stereolithography 3D-printed flow-through cell with a polyacrylamide (PAA) gel as its core. This gel needed to be coated with an agarose sheath because of its low viscosity. Drug release from hydroxypropyl methylcellulose-based implants containing either triamcinolone acetonide or fluorescein sodium was studied in the EFC using a schematic eye movement by the EyeMovementSystem (EyeMoS). For comparison, studies were performed in USP apparatus 4 and USP apparatus 7. Significantly slower drug release was observed in the PAA gel for both model drugs compared with the compendial methods. Drug release from fluorescein sodium-containing model implants was completed after 40 min in USP apparatus 4, whereas drug release in the gel-based EFC lasted 72 h. Drug release from triamcinolone acetonide-containing model implants was completed after 35 min in USP apparatus 4 and after 150 min in USP apparatus 7, whereas this was delayed until 96 h in the EFC. These results suggest that compendial release methods may overestimate the drug release rate in the human vitreous body. Using a gel-based in vitro release system such as the EFC may better predict drug release.

Background: Biosurfactants are surface-active molecules produced by different microorganisms and display a promising alternative to synthetically derived food emulsifiers. One of these biosurfactants, synthesized by Bacillus subtilis, is the lipopeptide surfactin, which composes a linear fatty acid and cyclic peptide moiety. This study explores the interfacial and emulsion forming properties of surfactin to further characterize its suitability as an O/W emulsifier in food formulations., Results: Surfactin revealed a high interfacial activity with a reduction of interfacial tension of 83.26 % to 4.21 ± 0.11 mN/m. O/W emulsions (c oil = 10 % w/w) were prepared by high-pressure homogenization, which yielded volume-based mean particle sizes below 1 μm already at low emulsifier concentrations of 0.01 % (w/w). Environmental stress experiments revealed that emulsions were stable between pH 6 to pH 9. Furthermore, neither phase separation nor extensive emulsion instability was observed with NaCl addition up to 0.5 M. However, CaCl 2 addition (> 3 mM) destabilized surfactin mediated emulsions. Finally, the main emulsion forming and stabilization effect of surfactin was related to its high interfacial activity and the high degree of electrostatic repulsion between the oil droplets (i.e. zeta-potential of up to -100 mV)., Conclusion: In comparison to other natural and synthetic emulsifiers, the results showed that surfactin is a strong candidate to form and stabilize O/W emulsions under the reported conditions., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

People are being encouraged to consume more plant-based foods to reduce the negative impacts of the modern food supply on human and global health. The food industry is therefore creating a new generation of plant-based products to meet this demand, including meat, fish, egg, milk, cheese, and yogurt analogs. The main challenge in this area is to simulate the desirable appearance, texture, flavor, mouthfeel, nutrition, and functionality of these products using healthy, affordable, and sustainable plant-derived ingredients, such as lipids, proteins, and carbohydrates. The molecular and physicochemical properties of plant-derived ingredients are very different from those of animal-derived ones. It is therefore critical to understand the fundamental attributes of plant-derived ingredients and how they can be assembled into structures resembling those found in animal products. This short review provides an overview of the current status of the scientific understanding of plant-based foods and highlights areas where further research is required. In particular, it focuses on the chemical, physical, and functional properties of plant ingredients; the processing operations that can be used to convert these ingredients into food products; and the science behind the creation of some common plant-based foods, namely meat, egg, and milk analogs.

Human induced pluripotent stem cell (iPSC) lines have been derived from four male patients with childhood attention-deficit hyperactivity disorder (ADHD). Children and adolescents between the ages 6 and 18 suffering from ADHD were recruited for this work. Isolated keratinocytes or peripheral blood mononuclear cells from the participants were reprogrammed into iPSCs using non-integrating Sendai virus to deliver the reprogramming factors Oct3/4, Sox2, Klf4 and c-Myc., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Ten human induced pluripotent stem cell (iPSC) lines have been derived from five healthy controls matched to a study including Attention-Deficit Hyperactivity Disorder patients (ADHD). Both female and male children and adolescents aged 6-18 years were recruited. Isolated keratinocyte cells from the participants were reprogrammed into iPSCs using non-integrating Sendai virus to deliver the reprogramming factors Oct3/4, Sox2, Klf4 and cMyc., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Objective: To investigate the value of standard [digital rectal examination (DRE), PSA] and advanced (mpMRI, prostate biopsy) clinical evaluation for prostate cancer (PCa) detection in contemporary patients with clinical bladder outlet obstruction (BOO) scheduled for Holmium laser enucleation of the prostate (HoLEP). Material and Methods: We retrospectively analyzed 397 patients, who were referred to our tertiary care laser center for HoLEP due to BOO between 11/2017 and 07/2020. Of those, 83 (20.7%) underwent further advanced clinical PCa evaluation with mpMRI and/or prostate biopsy due to elevated PSA and/or lowered PSA ratio and/or suspicious DRE. Logistic regression and binary regression tree models were applied to identify PCa in BOO patients. Results: An mpMRI was conducted in 56 (66%) of 83 patients and revealed PIRADS 4/5 lesions in 14 (25%) patients. Subsequently, a combined systematic randomized and MRI-fusion biopsy was performed in 19 (23%) patients and revealed in PCa detection in four patients (5%). A randomized prostate biopsy was performed in 31 (37%) patients and revealed in PCa detection in three patients (4%). All seven patients (9%) with PCa detection underwent radical prostatectomy with 29% exhibiting non-organ confined disease. Incidental PCa after HoLEP ( n = 76) was found in nine patients (12%) with advanced clinical PCa evaluation preoperatively. In univariable logistic regression analyses, PSA, fPSA ratio, and PSA density failed to identify patients with PCa detection. Conversely, patients with a lower International Prostate Symptom Score (IPSS) and PIRADs 4/5 lesion in mpMRI were at higher risk for PCa detection. In multivariable adjusted analyses, PIRADS 4/5 lesions were confirmed as an independent risk factor (OR 9.91, p = 0.04), while IPSS did not reach significance ( p = 0.052). Conclusion: In advanced clinical PCa evaluation mpMRI should be considered in patients with elevated total PSA or low fPSA ratio scheduled for BOO treatment with HoLEP. Patients with low IPSS or PIRADS 4/5 lesions in mpMRI are at highest risk for PCa detection. In patients with a history of two or more sets of negative prostate biopsies, advanced clinical PCa evaluation might be omitted., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Wenzel, Welte, Grossmann, Preisser, Theissen, Humke, Deuker, Bernatz, Gild, Ahyai, Karakiewicz, Bodelle, Kluth, Chun, Mandel and Becker.)

The modern food system is seeing a change in consumption patterns provoked by several drivers-including ethical, health, and environmental concerns-that are increasing the sales of meat analog foods. This change is accompanied by increased research and development activities in the area of plant-based meats. The aim of the present review is to describe methods that are being employed by scientists to analyze and characterize the properties of meat alternatives and to propose standardized methods that could be utilized in the future. In particular, methods to determine the proximate composition, microstructure, appearance, textural properties, water-holding properties, cooking resilience, and sensory attributes, of plant-based meat are given. The principles behind these methods are presented, their utility is critically assessed, and practical examples will be discussed. This article will help to guide further studies and to choose appropriate methods to assess raw materials, processes, products, and consumption behavior of meat analogs.

Goethite is a naturally anisotropic, antiferromagnetic iron oxide. Following its atomic structure, crystals grow into a fine needle shape that has interesting properties in a magnetic field. The needles align parallel to weak magnetic fields and perpendicular when subjected to high fields. We synthesized goethite nanorods with lengths between 200 nm and 650 nm in a two-step process. In a first step we synthesized precursor particles made of akaganeite (β-FeOOH) rods from iron(III)chloride. The precursors were then treated in a hydrothermal reactor under alkaline conditions with NaOH and polyvinylpyrrolidone (PVP) to form goethite needles. The aspect ratio was tunable between 8 and 15, based on the conditions during hydrothermal treatment. The orientation of these particles in a magnetic field was investigated by small angle X-ray scattering (SAXS). We observed that the field strength required to trigger a reorientation is dependent on the length and aspect ratio of the particles and could be shifted from 85 mT for the small particles to about 147 mT for the large particles. These particles could provide highly interesting magnetic properties to nanocomposites, that could then be used for sensing applications or membranes.

Attention-deficit hyperactivity disorder (ADHD) is one of the most common psychiatric neurodevelopmental disorders in children and adolescents. Although ADHD has been studied for nearly a century, the cause and pathophysiology of ADHD is yet largely unknown. However, findings from previous studies have resulted in the formation of a new hypothesis: Apart from the well-known multifactorial etiology of ADHD, recent evidence suggests that the interaction between genetic and environmental factors and especially Wnt- and mTOR-signaling pathways might have an important role in the pathophysiology of ADHD. The Wnt-signaling pathway is known to orchestrate cellular proliferation, polarity, and differentiation, and the mTOR pathway is involved in several significant processes of neurodevelopment and synaptic plasticity. As a result, dysregulations of these pathways in a time-dependent manner could lead to neurodevelopmental delays, resulting in ADHD phenotype. This review presents further evidence supporting our hypothesis by combining results from studies on ADHD and Wnt- or mTOR-signaling and the influence of genetics, methylphenidate treatment, Omega-3 supplementation, and stress.

Background: High-throughput sequencing (HTS) technologies are increasingly applied to analyse complex microbial ecosystems by mRNA sequencing of whole communities, also known as metatranscriptome sequencing. This approach is at the moment largely limited to prokaryotic communities and communities of few eukaryotic species with sequenced genomes. For eukaryotes the analysis is hindered mainly by a low and fragmented coverage of the reference databases to infer the community composition, but also by lack of automated workflows for the task., Results: From the databases of the National Center for Biotechnology Information and Marine Microbial Eukaryote Transcriptome Sequencing Project, 142 references were selected in such a way that the taxa represent the main lineages within each of the seven supergroups of eukaryotes and possess predominantly complete transcriptomes or genomes. From these references, we created an annotated microeukaryotic reference database. We developed a tool called TaxMapper for a reliably mapping of sequencing reads against this database and filtering of unreliable assignments. For filtering, a classifier was trained and tested on each of the following: sequences of taxa in the database, sequences of taxa related to those in the database, and random sequences. Additionally, TaxMapper is part of a metatranscriptomic Snakemake workflow developed to perform quality assessment, functional and taxonomic annotation and (multivariate) statistical analysis including environmental data. The workflow is provided and described in detail to empower researchers to apply it for metatranscriptome analysis of any environmental sample., Conclusions: TaxMapper shows superior performance compared to standard approaches, resulting in a higher number of true positive taxonomic assignments. Both the TaxMapper tool and the workflow are available as open-source code at Bitbucket under the MIT license: https://bitbucket.org/dbeisser/taxmapper and as a Bioconda package: https://bioconda.github.io/recipes/taxmapper/README.html .

Background: Chrysophytes are protist model species in ecology and ecophysiology and important grazers of bacteria-sized microorganisms and primary producers. However, they have not yet been investigated in detail at the molecular level, and no genomic and only little transcriptomic information is available. Chrysophytes exhibit different trophic modes: while phototrophic chrysophytes perform only photosynthesis, mixotrophs can gain carbon from bacterial food as well as from photosynthesis, and heterotrophs solely feed on bacteria-sized microorganisms. Recent phylogenies and megasystematics demonstrate an immense complexity of eukaryotic diversity with numerous transitions between phototrophic and heterotrophic organisms. The question we aim to answer is how the diverse nutritional strategies, accompanied or brought about by a reduction of the plasmid and size reduction in heterotrophic strains, affect physiology and molecular processes., Results: We sequenced the mRNA of 18 chrysophyte strains on the Illumina HiSeq platform and analysed the transcriptomes to determine relations between the trophic mode (mixotrophic vs. heterotrophic) and gene expression. We observed an enrichment of genes for photosynthesis, porphyrin and chlorophyll metabolism for phototrophic and mixotrophic strains that can perform photosynthesis. Genes involved in nutrient absorption, environmental information processing and various transporters (e.g., monosaccharide, peptide, lipid transporters) were present or highly expressed only in heterotrophic strains that have to sense, digest and absorb bacterial food. We furthermore present a transcriptome-based alignment-free phylogeny construction approach using transcripts assembled from short reads to determine the evolutionary relationships between the strains and the possible influence of nutritional strategies on the reconstructed phylogeny. We discuss the resulting phylogenies in comparison to those from established approaches based on ribosomal RNA and orthologous genes. Finally, we make functionally annotated reference transcriptomes of each strain available to the community, significantly enhancing publicly available data on Chrysophyceae., Conclusions: Our study is the first comprehensive transcriptomic characterisation of a diverse set of Chrysophyceaen strains. In addition, we showcase the possibility of inferring phylogenies from assembled transcriptomes using an alignment-free approach. The raw and functionally annotated data we provide will prove beneficial for further examination of the diversity within this taxon. Our molecular characterisation of different trophic modes presents a first such example., Competing Interests: Sven Rahmann is an Academic Editor for PeerJ.

Aquatic environments are heavily impacted by human activities including climate warming and the introduction of xenobiotics. Due to the application of silver nanoparticles as bactericidal agent the introduction of silver into the environment strongly has increased during the past years. Silver ions affect the primary metabolism of algae, in particular photosynthesis. Mixotrophic algae are an interesting test case as they do not exclusively rely on photosynthesis which may attenuate the harmful effect of silver. In order to study the effect of silver ions on mixotrophs, cultures of the chrysophyte Poterioochromonas malhamensis were treated in a replicate design in light and darkness with silver nitrate at a sub-lethal concentration. At five time points samples were taken for the identification and quantitation of proteins by mass spectrometry. In our analysis, relative quantitative protein mass spectrometry has shown to be a useful tool for functional analyses in conjunction with transcriptome reference sequences. A total of 3,952 proteins in 63 samples were identified and quantified, mapping to 4,829 transcripts of the sequenced and assembled transcriptome. Among them, 720 and 104 proteins performing various cellular functions were differentially expressed after eight days in light versus darkness and after three days of silver treatment, respectively. Specifically pathways of the energy and primary carbon metabolism were differentially affected by light and the utilization of expensive reactions hints to an energy surplus of P. malhamensis under light conditions. The excess energy is not invested in growth, but in the synthesis of storage metabolites. The effects of silver were less explicit, observable especially in the dark treatments where the light effect could not mask coinciding but weaker effects of silver. Photosynthesis, particularly the light harvesting complexes, and several sulphur containing enzymes were affected presumably due to a direct interference with the silver ions, mainly affecting energy supply., Competing Interests: The authors have declared that no competing interests exist.

Protists are perhaps the most lineage-rich of microbial lifeforms, but remain largely unknown. High-throughput sequencing technologies provide opportunities to screen whole habitats in depth and enable detailed comparisons of different habitats to measure, compare and map protistan diversity. Such comparisons are often limited by low sample numbers within single studies and a lack of standardisation between studies. Here, we analysed 232 samples from 10 sampling campaigns using a standardised PCR protocol and bioinformatics pipeline. We show that protistan community patterns are highly consistent within habitat types and geographic regions, provided that sample processing is standardised. Community profiles are only weakly affected by fluctuations of the abundances of the most abundant taxa and, therefore, provide a sound basis for habitat comparison beyond random short-term fluctuations in the community composition. Further, we provide evidence that distribution patterns are not solely resulting from random processes. Distinct habitat types and distinct taxonomic groups are dominated by taxa with distinct distribution patterns that reflect their ecology with respect to dispersal and habitat colonisation. However, there is no systematic shift of the distribution pattern with taxon abundance.

The functional properties of cutaneous afferent fibers were investigated 1-15 mo after nerve lesions, which allowed regeneration into denervated skin. After crushing or transection and resuturing the rat sural nerve, ongoing activity and responses to cold, heat, and mechanical stimuli presented to the denervated skin or to the nerve distal to the lesion were examined in 273 A-fibers and 211 C-fibers. Reinnervation of skin by A-fibers was largely complete by 1-4 mo after crushing but incomplete after transection and resuturing. A few A-fibers could be activated from the nerve trunk, even after 10-15 mo. Almost all regenerated A-fibers were mechanosensitive and about 6% were cold- or heat-sensitive. A few A-fibers had ongoing activity after nerve crush. Only 15-35% of C-fibers could be activated at 1-4 mo, but 60% were excited from the skin at 10-15 mo, when many also had receptive fields within the lesioned nerve. The remaining C-fibers had receptive fields only within the nerve trunk. Responses of both intraneural and intradermal endings of C-fibers could be classified into functional groups similar to those of C-fibers in control nerves to cutaneous stimuli. The frequency of afferent C-fibers with ongoing activity that were not highly cold sensitive was 45%. We conclude that the functional characteristics of afferent A- and C-fibers are expressed by regenerating nerve endings, even when they do not reinnervate their target tissue. The reinnervation of skin by afferent C-fibers is extremely slow and may never recover to normal.

Afferent A- and C-fibers regenerating into a nerve following peripheral nerve injury are exposed to inflammatory mediators released by Schwann cells, resident and invading macrophages, and other inflammatory cells. Here we tested the hypothesis that ongoing and evoked activity in these afferent fibers are enhanced by a mixture of inflammatory mediators [inflammatory soup (IS)] applied to the injured nerve. Using in vivo electrophysiology, regenerating afferent nerve fibers were studied 7-14 days after sural nerve crush lesion. The ectopic activity was studied before and

Isoprenylation of geranylgeranyl-pyrophosphate (GGPP) is critical for activation of small GTPases. We examined the roles of GGPP synthase (GGPPS) during the differentiation induced by the cell-to-cell contact in osteoblastic cell line MC3T3-E1 cells. We found that (1) both mRNA and protein expression of GGPPS was reduced with decrement of its activity during the differentiation, (2) GGOH, which is converted to GGPP in the cells, inhibited differentiation. These results suggest that the decrement of GGPP is critical for the cell-to-cell contact-induced differentiation, in which the down-regulation of GGPPS might be involved.