Seeking Solvation: Exploring the Role of Protein Hydration in Silk Gelation

The mechanism by which arthropods (e.g., spiders and many insects) can produce silk fibres from an aqueous protein (fibroin) solution has remained elusive, despite much scientific investigation. In this work, we used several techniques to explore the role of a hydration shell bound to the fibroin in native silk feedstock (NSF) from Bombyx mori silkworms. Small angle X-ray and dynamic light scattering (SAXS and DLS) revealed a coil size (radius of gyration or hydrodynamic radius) around 12 nm, providing considerable scope for hydration. Aggregation in dilute aqueous solution was observed above 65 °C, matching the gelation temperature of more concentrated solutions and suggesting that the strength of interaction with the solvent (i.e., water) was the dominant factor. Infrared (IR) spectroscopy indicated decreasing hydration as the temperature was raised, with similar changes in hydration following gelation by freezing or heating. It was found that the solubility of fibroin in water or aqueous salt solutions could be described well by a relatively simple thermodynamic model for the stability of the protein hydration shell, which suggests that the affected water is enthalpically favoured but entropically penalised, due to its reduced (vibrational or translational) dynamics. Moreover, while the majority of this investigation used fibroin from B. mori, comparisons with published work on silk proteins from other silkworms and spiders, globular proteins and peptide model systems suggest that our findings may be of much wider significance.

Closer to Nature: The Role of MSCs in Recreating the Microenvironment of the Hematopoietic Stem Cell Niche in vitro

<b><i>Background:</i></b> The stem cell niche in human bone marrow provides scaffolds, cellular frameworks and essential soluble cues to support the stemness of hematopoietic stem and progenitor cells (HSPCs). To decipher this complex structure and the corresponding cellular interactions, a number of in vitro model systems have been developed. The cellular microenvironment is of key importance, and mesenchymal stromal cells (MSCs) represent one of the major cellular determinants of the niche. Regulation of the self-renewal and differentiation of HSPCs requires not only direct cellular contact and adhesion molecules, but also various cytokines and chemokines. The C-X-C chemokine receptor type 4/stromal cell-derived factor 1 axis plays a pivotal role in stem cell mobilization and homing. As we have learned in recent years, to realistically simulate the physiological in vivo situation, advanced model systems should be based on niche cells arranged in a three-dimensional (3D) structure. By providing a dynamic rather than static setup, microbioreactor systems offer a number of advantages. In addition, the role of low oxygen tension in the niche microenvironment and its impact on hematopoietic stem cells need to be taken into account and are discussed in this review. <b><i>Summary:</i></b> This review focuses on the role of MSCs as a part of the bone marrow niche, the interplay between MSCs and HSPCs and the most important regulatory factors that need to be considered when engineering artificial hematopoietic stem cell niche systems. <b><i>Conclusion:</i></b> Advanced 3D model systems using MSCs as niche cells and applying microbioreactor-based technology are capable of simulating the natural properties of the bone marrow niche more closely than ever before.

Vertical Stratification of Insect Species Developing in Water-Filled Tree Holes

Forest ecosystems have a distinct vertical dimension, but the structuring of communities in this three-dimensional space is not well understood. Water-filled tree holes are natural microcosms structured in metacommunities. Here, we used these microcosms as model systems to analyze how insect communities and the occurrence and abundance of individual species are influenced by biotic and abiotic microhabitat characteristics, the vertical position of the tree hole, and stand-scale habitat availability. We found that both the characteristics of water-filled tree holes and their insect communities differ significantly between canopy and ground level. Individual insect species showed contrasting responses to the vertical position of the tree holes when important environmental factors at the stand and the tree-hole scale were considered. While some species, such as the mosquito Aedes geniculatus and the beetle Prionocyphon serricornis, decreased in abundance with increasing tree-hole height, the biting midge Dasyhelea sp., the non-biting midge Metriocnemus cavicola and the hoverfly Myiatropa florea increased in abundance. Our results suggest that vertical stratification in forests is most likely driven not only by variation in tree-hole microhabitat properties, i.e., niche separation, but also by individual species traits, such as adult dispersal propensity, food preferences and mating behavior of adult stages, and interspecific competition of larval stages. Therefore, communities of insect species developing in tree holes are likely structured by competition–colonization trade-offs predicted by metacommunity theory.

Memristive devices based on single ZnO nanowires - from material synthesis to neuromorphic functionalities

Memristive and resistive switching devices are considered promising building blocks for the realization of artificial neural networks and neuromorphic systems. Besides conventional top-down memristive devices based on thin films, resistive switching devices based on nanowires (NWs) have attracted great attention, not only for the possibility of going beyond current scaling limitations of the top-down approach, but also as model systems for the localization and investigation of the physical mechanism of switching. This work reports on the fabrication of memristive devices based on ZnO NWs, from NW synthesis to single NW-based memristive cell fabrication and characterization. The bottom-up synthesis of ZnO NWs was performed by low-pressure Chemical Vapor Deposition (LPCVD) according to a self-seeding Vapor-Solid (VS) mechanism on a Pt substrate over large scale (∼ cm2), without the requirement of previous seed deposition. The grown ZnO NWs are single crystalline with wurtzite crystal structure and are vertically aligned respect to the growth substrate. Single NWs were then contacted by means of asymmetric contacts, with an electrochemically active and an electrochemically inert electrode, to form NW-based electrochemical metallization memory (ECM) cells that show reproducible resistive switching behaviour and neuromorphic functionalities including short-term synaptic plasticity and Paired Pulse Facilitation (PPF). Besides representing building blocks for NW-based memristive and neuromorphic systems, these single crystalline devices can be exploited as model systems to study physicochemical processing underlaying memristive functionalities thanks to the high localization of switching events on the ZnO crystalline surface.

Bcl-XL but Not Bcl-2 Is a Potential Target in Medulloblastoma Therapy

Medulloblastoma (MB) is the most common solid tumour in children and, despite current treatment with a rather aggressive combination therapy, accounts for 10% of all deaths associated with paediatric cancer. Breaking the tumour cells’ intrinsic resistance to therapy-induced cell death should lead to less aggressive and more effective treatment options. In other tumour entities, this has been achieved by modulating the balance between the various pro- and anti-apoptotic members of the Bcl-2 family with small molecule inhibitors. To evaluate the therapeutic benefits of ABT-199 (Venetoclax), a Bcl-2 inhibitor, and ABT-263 (Navitoclax), a dual Bcl-XL/Bcl-2 inhibitor, increasingly more relevant model systems were investigated. Starting from established MB cell lines, progressing to primary patient-derived material and finally an experimental tumour system imbedded in an organic environment were chosen. Assessment of the metabolic activity (a surrogate readout for population viability), the induction of DNA fragmentation (apoptosis) and changes in cell number (the combined effect of alterations in proliferation and cell death induction) revealed that ABT-263, but not ABT-199, is a promising candidate for combination therapy, synergizing with cell death-inducing stimuli. Interestingly, in the experimental tumour setting, the sensitizing effect of ABT-263 seems to be predominantly mediated via an anti-proliferative and not a pro-apoptotic effect, opening a future line of investigation. Our data show that modulation of specific members of the Bcl-2 family might be a promising therapeutic addition for the treatment of MB.

The Evolution of Ovarian Carcinoma Subclassification

The phenotypically informed histotype classification remains the mainstay of ovarian carcinoma subclassification. Histotypes of ovarian epithelial neoplasms have evolved with each edition of the WHO Classification of Female Genital Tumours. The current fifth edition (2020) lists five principal histotypes: high-grade serous carcinoma (HGSC), low-grade serous carcinoma (LGSC), mucinous carcinoma (MC), endometrioid carcinoma (EC) and clear cell carcinoma (CCC). Since histotypes arise from different cells of origin, cell lineage-specific diagnostic immunohistochemical markers and histotype-specific oncogenic alterations can confirm the morphological diagnosis. A four-marker immunohistochemical panel (WT1/p53/napsin A/PR) can distinguish the five principal histotypes with high accuracy, and additional immunohistochemical markers can be used depending on the diagnostic considerations. Histotypes are further stratified into molecular subtypes and assessed with predictive biomarker tests. HGSCs have recently been subclassified based on mechanisms of chromosomal instability, mRNA expression profiles or individual candidate biomarkers. ECs are composed of the same molecular subtypes (POLE-mutated/mismatch repair-deficient/no specific molecular profile/p53-abnormal) with the same prognostic stratification as their endometrial counterparts. Although methylation analyses and gene expression and sequencing showed at least two clusters, the molecular subtypes of CCCs remain largely elusive to date. Mutational and immunohistochemical data on LGSC have suggested five molecular subtypes with prognostic differences. While our understanding of the molecular composition of ovarian carcinomas has significantly advanced and continues to evolve, the need for treatment options suitable for these alterations is becoming more obvious. Further preclinical studies using histotype-defined and molecular subtype-characterized model systems are needed to expand the therapeutic spectrum for women diagnosed with ovarian carcinomas.