Biological Research Research Articles

A multicenter high-quality data registry for advanced proton therapy approaches: the POWER registry

BackgroundPaucity and low evidence-level data on proton therapy (PT) represent one of the main issues for the establishment of solid indications in the PT setting. Aim of the present registry, the POWER registry, is to provide a tool for systematic, prospective, harmonized, and multidimensional high-quality data collection to promote knowledge in the field of PT with a particular focus on the use of hypofractionation.MethodsAll patients with any type of oncologic disease (benign and malignant disease) eligible for PT at the European Institute of Oncology (IEO), Milan, Italy, will be included in the present registry. Three levels of data collection will be implemented: Level (1) clinical research (patients outcome and toxicity, quality of life, and cost/effectiveness analysis); Level (2) radiological and radiobiological research (radiomic and dosiomic analysis, as well as biological modeling); Level (3) biological and translational research (biological biomarkers and genomic data analysis). Endpoints and outcome measures of hypofractionation schedules will be evaluated in terms of either Treatment Efficacy (tumor response rate, time to progression/percentages of survivors/median survival, clinical, biological, and radiological biomarkers changes, identified as surrogate endpoints of cancer survival/response to treatment) and Toxicity. The study protocol has been approved by the IEO Ethical Committee (IEO 1885). Other than patients treated at IEO, additional PT facilities (equipped with Proteus®ONE or Proteus®PLUS technologies by IBA, Ion Beam Applications, Louvain-la-Neuve, Belgium) are planned to join the registry data collection. Moreover, the registry will be also fully integrated into international PT data collection networks.

Construction of a glycosylation-mediated fluorescent biosensor for label-free measurement of site-specific 5-hydroxymethylcytosine in cancer cells with zero background signal

Background5-hydroxymethylcytosine (5hmC) as an epigenetic modification can regulate gene expression, and its abnormal level is related with various tumor invasiveness and poor prognosis. Nevertheless, the current methods for 5hmC assay usually involve expensive instruments/antibodies, radioactive risk, high background, laborious bisulfite treatment procedures, and non-specific/long amplification time. ResultsWe develop a glycosylation-mediated fluorescent biosensor based on helicase-dependent amplification (HDA) for label-free detection of site-specific 5hmC in cancer cells with zero background signal. The glycosylated 5hmC-DNA (5ghmC) catalyzed by β-glucosyltransferase (β-GT) can be cleaved by AbaSI restriction endonuclease to generate two dsDNA fragments with sticky ends. The resultant dsDNA fragments are complementary to the biotinylated probes and ligated by DNA ligases, followed by being captured by magnetic beads. After magnetic separation, the eluted ligation products act as the templates to initiate HDA reaction, generating abundant double-stranded DNA (dsDNA) products within 20 min. The dsDNA products are measured in a label-free manner with SYBR Green I as an indicator. This biosensor can measure 5hmC with a detection limit of 2.75 fM and a wide linear range from 1 × 10−14 to 1 × 10−8 M, and it can discriminate as low as 0.001% 5hmC level in complex mixture. Moreover, this biosensor can measure site-specific 5hmC in cancer cells, and distinguish tumor cells from normal cells. SignificanceThis biosensor can achieve a zero-background signal without the need of either 5hmC specific antibody or bisulfite treatment, and it holds potential applications in biological research and disease diagnosis.

Evaluation of Oil-Absorbing Film for Imprint Desorption Electrospray Ionization Mass Spectrometry Imaging (IDESI-MSI) on Biological Samples.

Imprint Desorption Electrospray Ionization Mass Spectrometry Imaging (IDESI-MSI) has proven to be a robust and reliable tool for chemically imaging biological samples such as fungi, animal tissues, and plants, but the choice of the imprint substrate is crucial. It must effectively transfer maximum amounts of species from the sample while preserving the original spatial distribution of detected molecules. In this study, we explored the potential of utilizing an oil-absorbing film, known for its soft nature and excellent lipophilicity, as an imprint substrate for IDESI-MSI on biological samples. To assess the transfer efficiency of the amounts of molecules and molecular patterns, we conducted experiments using mouse brain tissue. The result shows that more than 90% of the analytes can be transferred to the oil-absorbing film from the original tissue. A comparison of IDESI-MSI results between the oil-absorbing film and the original tissue demonstrates the material's capability to transfer most molecules from the original tissue and retain images of different analytes with high spatial fidelity. We extended our investigation to plant imaging, where we applied IDESI-MSI to a cross-section of okra. The oil-absorbing film exhibited promise in this context as well. These findings suggest that IDESI-MSI utilizing the oil-absorbing film holds potential across various research fields, including biological metabolism, chemistry, and clinical research, making this technique widely applicable.

Transition-Metal-Catalyzed C–H Activation Reactions for the Creation and Modification of Organic Fluorophores

AbstractOrganic fluorophores have consistently garnered significant interest owing to their widespread application across various multidisciplinary research fields. In the realm of biological research, these organic fluorophores find extensive use in diverse applications such as molecular imaging, DNA sequencing, drug discovery, and biosensors. Remarkably, in recent times, organic fluorescent molecules have emerged as pivotal elements in the advancement of organic electronics. Across several reaction pathways developed for constructing and modifying organic fluorophores, transition-metal-catalyzed C–H activation reactions have come across as a dependable and step-economical approach. In this review we discuss various transition-metal-catalyzed C–H activation-based approaches that have been employed to create and modify organic fluorescent molecules which find applications in multidisciplinary research areas.1 Introduction2 Basic Reactions for the Creation of Organic Fluorophores3 Merits and Drawbacks of Classical Reactions in the Creation and Modification of Organic Fluorophores4 C–H Activation/Functionalization Reactions5 C–H Activation Pathways in the Creation and Modification of Organic­ Fluorophores5.1 Electrophilic C–H Activation Reactions5.2 Heteroatom-Directed C–H Activation Reactions6 Conclusion

Cysteine-Specific Multifaceted Bioconjugation of Peptides and Proteins Using 5-Substituted 1,2,3-Triazines.

Peptide and protein postmodification have gained significant attention due to their extensive impact on biomolecule engineering and drug discovery, of which cysteine-specific modification strategies are prominent due to their inherent nucleophilicity and low abundance. Herein, the study introduces a novel approach utilizing multifunctional 5-substituted 1,2,3-triazine derivatives to achieve multifaceted bioconjugation targeting cysteine-containing peptides and proteins. On the one hand, this represents an inaugural instance of employing 1,2,3-triazine in biomolecular-specific modification within a physiological solution. On the other hand, as a powerful combination of precision modification and biorthogonality, this strategy allows for the one-pot dual-orthogonal functionalization of biomolecules utilizing the aldehyde group generated simultaneously. 1,2,3-Triazine derivatives with diverse functional groups allow conjugation to peptides or proteins, while bi-triazines enable peptide cyclization and dimerization. The examination of the stability of bi-triazines revealed their potential for reversible peptide modification. This work establishes a comprehensive platform for identifying cysteine-selective modifications, providing new avenues for peptide-based drug development, protein bioconjugation, and chemical biology research.

Lung Organoids: Systematic Review of Recent Advancements and its Future Perspectives.

Organoids are essentially an in vitro (lab-grown) three-dimensional tissue culture system model that meticulously replicates the structure and physiology of human organs. A few of the present applications of organoids are in the basic biological research area, molecular medicine and pharmaceutical drug testing. Organoids are crucial in connecting the gap between animal models and human clinical trials during the drug discovery process, which significantly lowers the time duration and cost associated with each stage of testing. Likewise, they can be used to understand cell-to-cell interactions, a crucial aspect of tissue biology and regeneration, and to model disease pathogenesis at various stages of thedisease. Lung organoids can be utilized to explore numerous pathophysiological activities of a lung since they share similarities with its function. Researchers have been trying to recreate the complex nature of the lung by developing various "Lung organoids" models.This article is a systematic review of various developments of lung organoids and their potential progenitors. It also covers the in-depth applications of lung organoids for the advancement of translational research. The review discusses the methodologies to establish different types of lung organoids for studying the regenerative capability of the respiratory system and comprehending various respiratory diseases.Respiratory diseases are among the most common worldwide, and the growing burden must be addressed instantaneously.Lung organoids along with diverse bio-engineering tools and technologies will serve as a novel model for studying thepathophysiology of various respiratory diseasesand fordrug screening purposes.

Revolutionizing bone tumor management: cutting-edge breakthroughs in limb-saving treatments.

Limb salvage surgery has revolutionized the approach to bone tumors in orthopedic oncology, steering away from historical amputations toward preserving limb function and enhancing patient quality of life. This transformative shift underscores the delicate balance between tumor eradication and optimal postoperative function. Primary and metastatic bone tumors present challenges in early detection, differentiation between benign and malignant tumors, preservation of function, and the risk of local recurrence. Conventional methods, including surgery, radiation therapy, chemotherapy, and targeted therapies, have evolved with a heightened focus on personalized medicine. A groundbreaking development in limb salvage surgery is the advent of 3D-printed patient-specific implants, which significantly enhance anatomical precision, stability, and fixation. These implants reduce soft tissue disruption and the associated risks, fostering improved osseointegration and correction of deformities for a more natural and functional postoperative outcome. Biological and molecular research has reshaped the understanding of bone tumors, guiding surgical interventions with advancements such as genomic profiling, targeted intraoperative imaging, precision targeting of molecular pathways, and immunotherapy tailored to individual tumor characteristics. In the realm of imaging technologies, MRI, CT scans, and intraoperative navigation systems have redefined preoperative planning, minimizing collateral damage and optimizing outcomes through accurate resections. Postoperative rehabilitation plays a crucial role in restoring function and improving the quality of life. Emphasizing early mobilization, effective pain management, and a multidisciplinary approach, rehabilitation addresses the physical, psychological, and social aspects of recovery. Looking ahead, future developments may encompass advanced biomaterials, smart implants, AI algorithms, robotics, and regenerative medicine. Challenges lie in standardization, cost-effectiveness, accessibility, long-term outcome assessment, mental health support, and fostering global collaboration. As research progresses, limb salvage surgery emerges not just as a preservation tool but as a transformative approach, restoring functionality, resilience, and hope in the recovery journey. This review summarizes the recent advances in limb salvage therapy for bone tumors over the past decade.

Estimation of Temperature and Salinity from Marine Seismic Data—A Two-Step Approach

Ocean-water temperature and salinity are two vital properties that are required for weather-, climate-, and marine biology-related research. These properties are usually measured using disposable instruments at sparse locations, typically from tens to hundreds of kilometers apart. Laterally interpolating these sparse measurements provides smooth temperature and salinity distributions within the oceans, although they may not be very accurate. Marine seismic data, on the other hand, show visible reflections within the water-column which are primarily controlled by subtle sound-speed variations. Because these variations are functions of the temperature, salinity, and pressure, estimating sound-speed from marine seismic data and relating them to temperature and salinity have been attempted in the past. These seismically derived properties are of much higher lateral resolution (less than 25 m) than the sparse measurements and can be potentially used for climate and marine biology research. Estimating sound-speeds from seismic data, however, requires running iterative seismic inversions, which need a good initial model. Currently practiced ways to generate this initial model are computationally challenging, labor-intensive, and subject to human error and bias. In this research, we outline an automated method to generate the initial model which is neither computational and labor-intensive nor prone to human errors and biases. We also use a two-step process of, first, estimating the sound-speed from seismic inversion data and then estimating the salinity and temperature. Furthermore, by applying this method to real seismic data, we demonstrate the feasibility of our approach and discuss how the use of machine learning can further improve the computational efficiency of the method and make an impact on the future of climate modeling, weather prediction, and marine biology research.

Quality and applicability of cadaveric donor eyes for molecular biology research: An Indian experience.

Human ocular tissue banking plays an important part in the advancement of translational research for identifying the molecular processes involved in disease etiology and pathogenesis. Timely obtaining a good-quality ocular tissue from a cadaveric donor is exceedingly difficult, especially in remote areas, with a variable transportation time (within 12-24 h), raising concerns about RNA quality and its subsequent applications. Therefore, we assessed the utility of retinal tissues from cadaver donor and enucleated eyes based on the RNA quality and gene expression by real-time polymerase chain reaction (PCR). Prospective study. Retina tissues were separated from the donor/enucleated eyes received in the eye bank within 24 h of death (n = 15) and within an hour from OR (n = 3), respectively, and stored immediately at -80 degree. RNA was isolated using trizol, and the quantity and quality were assessed using Qubit and agarose gel electrophoresis, respectively. QPCR was performed for measuring the expression of different retinal-specific genes. The cellular viability of the retina was assessed by establishing explant primary cell cultures. The data were calculated as an average of normalised Ct values ± standard error of the mean. RNA obtained from cadaveric tissues despite being partially degraded showed a uniform strong gene expression of several retinal-specific genes such as PAX6, RHO, TUBB3, CRX, and ALDH1L1. The primary cultures established from cadaveric tissues showed viable cells. The cadaver donor tissues collected within 24 hours of death can be effectively utilized for gene expression profiling.

Home Sweet Home: Setting the Best Thriving Conditions for the Ad Hoc Engineered Microbial Consortium in the Zero Mile System

Wastewaters from household appliances, such as dishwashers and washing machines, are an untapped resource of recoverable water and/or nutrients. The Zero Mile system has been developed to reuse/upcycle dishwasher wastewaters through bioremediation activity carried out by an ad hoc engineered phototrophic/heterotrophic microbial consortium. The choice of both suitable microorganisms for engineering consortia and detailed knowledge on their structure, behaviour and interaction are essential to optimising consortium culture conditions and drive the biofilter container design (structure and topology). To these aims, the effect of abiotic conditions (i.e., irradiance, pH and organic load) on the microbial consortium growth and its capability to survive and thrive in different dishwasher wastewater dilutions have been evaluated. At the same time, the crucial interplay between biological and design research has allowed us to define the characteristics of the biofilter container and plan its development for the industrial application of the Zero Mile system, bringing sustainability benefits as it moves household wastewater from a traditional linear model to a more sustainable, circular approach.

Bacterial chemotaxis control process analysis with SysML

AbstractThis paper looks at the bacteria chemotaxis control process utilizing the System Modeling Language (SysML) to leverage well‐defined and proven engineering tools for architecting, analyzing, and refining complex systems. It proposes a new methodology called reverse‐engineering object‐oriented systems engineering method (RE‐OOSEM) that converts descriptive biology research information into descriptive systems engineering information. It utilizes SysML and model‐based systems engineering (MBSE) to capture system architecture from biological system knowledge and inputs them into systems engineering tools. From an engineering point of view, this allows greater insight into how biological systems operate and suggests how much model detail is required to uncover a top‐down system understanding. RE‐OOSEM methodology guides the SysML chemotaxis control capture process. SysML syntax is used instead of biological syntax to facilitate biological chemotaxis control system analysis from an engineered system point of view. The model can act as a scaffolding to help uncover system function, the relationships of system components and processes, and bioinformatic phenotype and genotype correlation. An executable MathWorks Stateflow chemotaxis control process model based on the SysML architectural model is included. The results show the following engineering perspective observations. (1) Several control components are not dedicated but are available and utilized when needed. (2) Individual chemoreceptors act together as a sensor array. (3) Phosphate groups act as a signaling mechanism. (4) Methylation via CH3 groups of the chemoreceptor results in sensitivity adaptation. (5) Closed‐loop control collaboratively utilizes ligand bonding, phosphorylation, and methylation. (6) Timing relationships of the control subprocesses give insight into the system's architecture.

Unique fluorescent probe for the recognition of late apoptosis via translocation from plasma membrane to nucleus

Cell apoptosis is a crucial physiological process playing central roles in key biological and pathological activities. However, the current fluorescent probes for the detection of late apoptosis were “off–on” probes, which were facilely interfered by false positive signals caused by inhomogeneous staining and other factors. Herein, a unique fluorescent probe (NPn) discriminating late apoptosis from early apoptosis and heathy status with two different sets of fluorescent signals have been prepared, to overcome the possible false positive signals. NPn was designed impermeable to biomembranes and simultaneously with high affinity to DNA/RNA, which localized on the plasma membranes of living and early apoptotic cells, while relocated to the nucleus in late apoptotic cells. The hydrophilic amine unit and small ion radius were responsive for its membrane impermeability, which was confirmed with two control molecules without amine group. Using the probe, we have successfully evaluated the cell apoptosis induced by ultraviolet irradiation, rotenone, colchicine, and paclitaxel, demonstrating its potential application in biological researches.

The Transthyretin Protein and Amyloidosis – an Extraordinary Chemical Biology Platform

AbstractThe amyloidoses are diseases caused by accumulation of amyloid fibrils from over 40 different human misfolded proteins in various organs of the body depending on precursor protein. Amyloidogenesis is a self‐perpetuating reaction with deleterious consequences causing degeneration in cells and organs where depositions occur. Transthyretin, TTR, is an amyloidogenic protein causing sporadic disease from the wild‐type protein during aging and from numerous different autosomal dominant familial mutations at earlier ages depending on the sequence of the hereditary variant. Until recently the disease process was poorly understood, and therapies were scarce. Over the past decades, spurred by clinical data, using chemical biology research, the mechanisms of TTR production and misfolding have been elucidated affording almost complete coverage of the TTR amyloidogenesis pathway to be targeted. This translational science success has provided a plethora of therapeutic options for the TTR amyloidoses providing an inspiring example for success in previously intractable diseases.

Hybrid representation learning for cognitive diagnosis in late-life depression over 5 years with structural MRI

Late-life depression (LLD) is a highly prevalent mood disorder occurring in older adults and is frequently accompanied by cognitive impairment (CI). Studies have shown that LLD may increase the risk of Alzheimer’s disease (AD). However, the heterogeneity of presentation of geriatric depression suggests that multiple biological mechanisms may underlie it. Current biological research on LLD progression incorporates machine learning that combines neuroimaging data with clinical observations. There are few studies on incident cognitive diagnostic outcomes in LLD based on structural MRI (sMRI). In this paper, we describe the development of a hybrid representation learning (HRL) framework for predicting cognitive diagnosis over 5 years based on T1-weighted sMRI data. Specifically, we first extract prediction-oriented MRI features via a deep neural network, and then integrate them with handcrafted MRI features via a Transformer encoder for cognitive diagnosis prediction. Two tasks are investigated in this work, including (1) identifying cognitively normal subjects with LLD and never-depressed older healthy subjects, and (2) identifying LLD subjects who developed CI (or even AD) and those who stayed cognitively normal over five years. We validate the proposed HRL on 294 subjects with T1-weighted MRIs from two clinically harmonized studies. Experimental results suggest that the HRL outperforms several classical machine learning and state-of-the-art deep learning methods in LLD identification and prediction tasks.

The horizontal ladder test (HLT) protocol: a novel, optimized, and reliable means of assessing motor coordination in Sus scrofa domesticus.

Pigs can be an important model for preclinical biological research, including neurological diseases such as Alcohol Use Disorder. Such research often involves longitudinal assessment of changes in motor coordination as the disease or disorder progresses. Current motor coordination tests in pigs are derived from behavioral assessments in rodents and lack critical aspects of face and construct validity. While such tests may permit for the comparison of experimental results to rodents, a lack of validation studies of such tests in the pig itself may preclude the drawing of meaningful conclusions. To address this knowledge gap, an apparatus modeled after a horizontally placed ladder and where the height of the rungs could be adjusted was developed. The protocol that was employed within the apparatus mimicked the walk and turn test of the human standardized field sobriety test. Here, five Sinclair miniature pigs were trained to cross the horizontally placed ladder, starting at a rung height of six inches and decreasing to three inches in one-inch increments. It was demonstrated that pigs can reliably learn to cross the ladder, with few errors, under baseline/unimpaired conditions. These animals were then involved in a voluntary consumption of ethanol study where animals were longitudinally evaluated for motor coordination changes at baseline, 2.5, 5, 7.5, and 10% ethanol concentrations subsequently to consuming ethanol. Consistent with our predictions, relative to baseline performance, motor incoordination increased as voluntary consumption of escalating concentrations of ethanol increased. Together these data highlight that the horizontal ladder test (HLT) test protocol is a novel, optimized and reliable test for evaluating motor coordination as well as changes in motor coordination in pigs.

Mesenchyme instructs growth while epithelium directs branching in the mouse mammary gland.

The mammary gland is a unique organ that undergoes dynamic alterations throughout a female's reproductive life, making it an ideal model for developmental, stem cell and cancer biology research. Mammary gland development begins in utero and proceeds via a quiescent bud stage before the initial outgrowth and subsequent branching morphogenesis. How mammary epithelial cells transit from quiescence to an actively proliferating and branching tissue during embryogenesis and, importantly, how the branch pattern is determined remain largely unknown. Here, we provide evidence indicating that epithelial cell proliferation and onset of branching are independent processes, yet partially coordinated by the Eda signaling pathway. Through heterotypic and heterochronic epithelial-mesenchymal recombination experiments between mouse mammary and salivary gland tissues and ex vivo live imaging, we demonstrate that unlike previously concluded, the mode of branching is an intrinsic property of the mammary epithelium whereas the pace of growth and the density of ductal tree are determined by the mesenchyme. Transcriptomic profiling and ex vivo and in vivo functional studies in mice disclose that mesenchymal Wnt/ß-catenin signaling, and in particular IGF-1 downstream of it critically regulate mammary gland growth. These results underscore the general need to carefully deconstruct the different developmental processes producing branched organs.

Revealing mitf functions and visualizing allografted tumor metastasis in colorless and immunodeficient Xenopus tropicalis

Transparent immunodeficient animal models not only enhance in vivo imaging investigations of visceral organ development but also facilitate in vivo tracking of transplanted tumor cells. However, at present, transparent and immunodeficient animal models are confined to zebrafish, presenting substantial challenges for real-time, in vivo imaging studies addressing specific biological inquiries. Here, we employed a mitf−/−/prkdc−/−/il2rg−/− triple-knockout strategy to establish a colorless and immunodeficient amphibian model of Xenopus tropicalis. By disrupting the mitf gene, we observed the loss of melanophores, xanthophores, and granular glands in Xenopus tropicalis. Through the endogenous mitf promoter to drive BRAFV600E expression, we confirmed mitf expression in melanophores, xanthophores and granular glands. Moreover, the reconstruction of the disrupted site effectively reinstated melanophores, xanthophores, and granular glands, further highlighting the crucial role of mitf as a regulator in their development. By crossing mitf−/− frogs with prkdc−/−/il2rg−/− frogs, we generated a mitf−/−/prkdc−/−/il2rg−/−Xenopus tropicalis line, providing a colorless and immunodeficient amphibian model. Utilizing this model, we successfully observed intravital metastases of allotransplanted xanthophoromas and migrations of allotransplanted melanomas. Overall, colorless and immunodeficient Xenopus tropicalis holds great promise as a valuable platform for tumorous and developmental biology research.

Single-Nucleotide-Specific Lipidic Nanoflares for Precise and Visible Detection of KRAS Mutations via Toehold-Initiated Self-Priming DNA Polymerization.

Accurate identification of single-nucleotide mutations in circulating tumor DNA (ctDNA) is critical for cancer surveillance and cell biology research. However, achieving precise and sensitive detection of ctDNAs in complex physiological environments remains challenging due to their low expression and interference from numerous homologous species. This study introduces single-nucleotide-specific lipidic nanoflares designed for the precise and visible detection of ctDNA via toehold-initiated self-priming DNA polymerization (TPP). This system can be assembled from only a single cholesterol-conjugated multifunctional molecular beacon (MMB) via hydrophobicity-mediated aggregation. This results in a compact, high-density, and nick-hidden arrangement of MMBs on the surface of lipidic micelles, thereby enhancing their biostability and localized concentrations. The assay commences with the binding of frequently mutated regions of ctDNA to the MMB toehold domain. This domain is the proximal holding point for initiating the TPP-based strand-displacement reaction, which is the key step in enabling the discrimination of single-base mutations. We successfully detected a single-base mutation in ctDNA (KRAS G12D) in its wild-type gene (KRAS WT), which is one of the most frequently mutated ctDNAs. Notably, coexisting homologous species did not interfere with signal transduction, and small differences in these variations can be visualized by fluorescence imaging. The limit of detection was as low as 10 amol, with the system functioning well in physiological media. In particular, this system allowed us to resolve genetic mutations in the KRAS gene in colorectal cancer, suggesting its high potential in clinical diagnosis and personalized medicine.

Hydroponics in physiological studies of trace element tolerance and accumulation in plants focussing on metallophytes and hyperaccumulator plants

Abstract Background When studying metallophytes and hyperaccumulator plants, it is often desired to assess the level of tolerance of a specific trace metal/metalloid in a putative tolerant species, to determine root and shoot accumulation of the trace metal/metalloid of interest, or to establish whether a trace metal/metalloid has an essential function. The use of hydroponics has proven to be a powerful tool in answering such questions in relation to the physiological regulation of metal/metalloids in plants. Carefully designing experiments requires considering nutrient solution formulation, dose rate regime, and environmental conditions, but this is often overlooked. Aims This review aims to bring together key information for hydroponics studies in physiological, evolutionary, and genetics/molecular biological research of trace metal/metalloid tolerance and accumulation in plants, focussing on metallophytes and hyperaccumulator plants. Conclusions It is not possible to define a ‘universal’ nutrient solution that is both sufficient and non-toxic for all plants, although it is often possible, dependent on plant species under study and the research question to be addressed, to ‘adapt’ commonly used ‘standard formulations’. Well-designed and executed hydroponics experiments can yield powerful insights in the regulation of essential and toxic metal/metalloid trace elements, and this extends far beyond hyperaccumulator plants.

From static to dynamic: The influence of mechanotransduction on skin equivalents analyzed by bioimaging and RNAseq

In this study, we explore the impact of mechanical stimuli on skin models using an innovative skin-on-a-chip platform, addressing the limitations of conventional transwell-cultured skin equivalents. This platform facilitates cyclic mechanical stimulation through compression and stretching, combined with automated media perfusion. Our findings, using bioimaging and bulk RNA sequencing, reveal increased expression of Keratin 10 and Keratin 14, indicating enhanced skin differentiation and mechanical integrity. The increase in desmosomes and tight junctions, observed through Claudin-1 and Desmoplakin 1 & 2 analysis, suggests improved keratinocyte differentiation due to mechanical stimulation. Gene expression analyses reveal a nuanced regulatory response, suggesting a potential connection to the Hippo pathway, indicative of a significant cellular reaction to mechanical stimuli. The results show the important influence of mechanical stimulation on skin model integrity and differentiation, demonstrating the potential of our microfluidic platform in advancing skin biology research and pharmaceutical testing.