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Pyrethroids are chemical insecticides that are widely used to control pests. Entomopathogenic fungi are considered environmentally safe alternatives to these compounds. Pyrethroids and entomopathogenic fungi not only co-exist in the environment but can also be applied together in pest control. They are often found in contact with each other, and thus, it seems important to understand their interactions at the cellular level. In this study, we analyzed whether pyrethroids could influence the phospholipid profile of Beauveria bassiana and whether membrane changes are one of the mechanisms by which these fungi adapt to unfavorable environmental conditions. The results of our study revealed that pyrethroids changed the phospholipid profile and increased the cell membrane permeability of B. bassiana, which enabled them to enter and accumulate within the fungal cells, resulting in oxidative stress. Pyrethroids influenced the amount of neutral lipids, caused a decrease in sodium content, and also temporarily lowered the level of the secondary metabolite oosporein in the studied fungi. These findings indicate that the effect of pyrethroids on entomopathogenic fungi may be more complex than originally thought and that lipidomic studies can aid in fully understanding the influence of these chemicals on the mentioned group of fungi.

Pre‐emergence herbicides are taken up by seeds before germination and by roots, hypocotyls, cotyledons, coleoptiles or leaves before emergence, whereas post‐emergence herbicides are taken up primarily by foliage and stems. Most modern pre‐emergence herbicides are lipophilic, but post‐emergence herbicides may be lipophilic or hydrophilic. The metabolic conversion of herbicides to inactive or active metabolites after plant uptake is of major importance for some compound classes. Several herbicides are proherbicides as for example some acetyl‐coenzyme A carboxylase (ACCase)‐inhibitors. The physicochemical characteristics of proherbicides and herbicides are usually unrelated. A major role can be attributed to the site of action at a cellular level. A great number of herbicides such as photosystem II (PS II)‐inhibitors, protoporphyrinogen oxidase (PPO)‐inhibitors or carotenoid biosynthesis inhibitors require light for activity. Others, such as cellulose‐biosynthesis and mitotic inhibitors seem to be primarily active in belowground organs. Several lipophilic barriers against the uptake of xenobiotics exist in aboveground and belowground plant parts. The relevance of these barriers needs, however, further clarification. Uptake and translocation models are valuable tools for the explanation of the potential movement of compounds. Many factors other than uptake and translocation have, however, to be considered for the design of herbicides. For post‐emergence herbicides, ultraviolet (UV) light stability, stability in formulations, and mixability with other agrochemicals have to be kept in mind while, in addition to the aforementioned factors soil interaction plays a major role for pre‐emergence herbicides. In our opinion, general physicochemical characteristics of pre‐ or post‐emergence herbicides do, unfortunately not exist yet. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., A high number of parameters influence the fate of herbicides. General characteristics of pre‐ or post‐emergence herbicides can therefore not be defined yet.

The architecture of the mouse mammary gland is highly dynamic and constantly remodeled during pubertal development and estrous cycle-driven sprouting and regression of alveolar side branches. During each of these developmental stages, turnover is driven by distinct subsets of mammary epithelial cells. Extensive previous research has shed light on the unique morphological and cell biological characteristics of each stage. However, technological shortcomings failed to capture the dynamics and single-cell contributions to mammary remodeling. Here, we developed in vivo imaging strategies to follow the same mammary ducts over time and quantify the dynamics of mammary gland growth and remodeling from single-cell level to organ scale. Using a combination of intravital microscopy and genetic reporter systems we show how proliferative heterogeneity drives ductal morphogenesis during different developmental stages. To visualize pubertal growth at the cellular level, we performed long-term time-lapse imaging of extending terminal end buds through a mammary imaging window. We show that single-cells within the terminal end buds are extremely motile and continuously exchange position whilst the duct is elongating. To visualize short-term remodeling in the adult mammary gland at the single cell level, we performed multi-day intravital imaging in photoconvertible Kikume Green-Red mice and fluorescent ubiquitination-based cell cycle indicator mice. We demonstrate that the contribution of single-cells to estrous-driven remodeling is highly variable between cells in the same micro-environment. To assess the effects of this dynamic proliferative contribution on the long-term stability of tissue architecture, we developed a repeated skin flap method to assess mammary gland morphology by intravital microscopy over extended time spans for up to six months. Interestingly, in contrast to the short-term dynamic remodeling, the long-term morphology of the mammary gland remains remarkably stable. Together, our tool box of imaging strategies allows to identify and map transient and continuing dynamics of single cells to the architecture of the mammary gland. ispartof: JOURNAL OF MAMMARY GLAND BIOLOGY AND NEOPLASIA vol:26 issue:1 pages:9-27 ispartof: location:United States status: published

Analysis of growth and death kinetics at single-cell resolution is a key step in understanding the complexity of the nonreplicating growth phenotype of the bacterial pathogen Mycobacterium tuberculosis. Here, we developed a single-cell-resolution microfluidic mycobacterial culture device that allows time-lapse microscopy-based long-term phenotypic visualization of the live replication dynamics of mycobacteria. This technology was successfully applied to monitor the real-time growth dynamics of the fast-growing model strain Mycobacterium smegmatis (M. smegmatis) while subjected to drug treatment regimens during continuous culture for 48 h inside the microfluidic device. A clear morphological change leading to significant swelling at the poles of the bacterial membrane was observed during drug treatment. In addition, a small subpopulation of cells surviving treatment by frontline antibiotics was observed to recover and achieve robust replicative growth once regular culture media was provided, suggesting the possibility of identifying and isolating nonreplicative mycobacteria. This device is a simple, easy-to-use, and low-cost solution for studying the single-cell phenotype and growth dynamics of mycobacteria, especially during drug treatment.

Cancer stem cells (CSCs) are a subpopulation of cancer cells with functions similar to those of normal stem cells. Although few in number, they are capable of self-renewal, unlimited proliferation, and multi-directional differentiation potential. In addition, CSCs have the ability to escape immune surveillance. Thus, they play an important role in the occurrence and development of tumors, and they are closely related to tumor invasion, metastasis, drug resistance, and recurrence after treatment. Therefore, specific targeting of CSCs may improve the efficiency of cancer therapy. A series of corresponding promising therapeutic strategies based on CSC targeting, such as the targeting of CSC niche, CSC signaling pathways, and CSC mitochondria, are currently under development. Given the rapid progression in this field and nanotechnology, drug delivery systems (DDSs) for CSC targeting are increasingly being developed. In this review, we summarize the advances in CSC-targeted DDSs. Furthermore, we highlight the latest developmental trends through the main line of CSC occurrence and development process; some considerations about the rationale, advantages, and limitations of different DDSs for CSC-targeted therapies were discussed., Graphical abstract Emerging CSCs-targeted drug delivery systems for efficient cancer therapy following the main line of CSCs occurrence and development process from the whole to the part and from the outside to the inside.Image 1

Abstract Diabetes mellitus is referred as common metabolic abnormalities characterized as hyperglycemia, mainly caused due to insufficient production of insulin at cellular level or/and defects in insulin action. Such an endocrine disorder is responsible for serious health problems and its worldwide prevalence is rapidly increasing. Common management of diabetes by oral administration of drugs without creating any side effects is still considered a challenging task and increasing cost of conventional medicine in developing countries is another matter of concern. To address these issues, traditional preparations of herbal plant extracts in the form of medicines already gained immense attention. Swertia chirayita is one among such plants which is known for its hypoglycemic potential. Numerous chemical constituents with promising pharmacological properties have been identified from such plant extracts but still, such compounds have not been well characterized, specifically against human application. Hence, more research efforts are necessitated to understand exact mechanism of such compounds and to develop overall safety of such plant formulations. Present review clearly represents antidiabetic properties of Swertia chirayita extract, strategies to be taken to strengthen its safety application on humans and biotechnological interventions that ensure conservation of such endangered species to promote its future application in modern medicine. Graphic Abstract

Living cells interpret a variety of signals in different contexts to elucidate functional responses. While the understanding of signalling molecules, their respective receptors and response at the gene transcription level have been relatively well-explored, how exactly does a single cell interpret a plethora of time-varying signals? Furthermore, how their subsequent responses at the single cell level manifest in the larger context of a developing tissue is unknown. At the same time, the biophysics and chemistry of how receptors are trafficked through the complex dynamic transport network between the plasma membrane–endosome–lysosome–Golgi–endoplasmic reticulum are much more well-studied. How the intracellular organisation of the cell and inter-organellar contacts aid in orchestrating trafficking, as well as signal interpretation and modulation by the cells are beginning to be uncovered. In this review, we highlight the significant developments that have strived to integrate endosomal trafficking, signal interpretation in the context of developmental biology and relevant open questions with a few chosen examples. Furthermore, we will discuss the imaging technologies that have been developed in the recent past that have the potential to tremendously accelerate knowledge gain in this direction while shedding light on some of the many challenges.

Aging and circadian rhythms are two biological processes that affect an organism, although at different time scales. Nevertheless, due to the overlap of their actions, it was speculated that both interfere or interact with each other. However, to address this question, a much deeper insight into these processes is necessary, especially at the cellular level. New methods such as single-cell RNA-sequencing (scRNA-Seq) have the potential to close this gap in our knowledge. In this review, we analyze applications of scRNA-Seq from the aging and circadian rhythm fields and highlight new findings emerging from the analysis of single cells, especially in humans or rodents. Furthermore, we judge the potential of scRNA-Seq to identify common traits of both processes. Overall, this method offers several advantages over more traditional methods analyzing gene expression and will become an important tool to unravel the link between these biological processes.

Actively regulated symmetry breaking, which is ubiquitous in biological cells, underlies phenomena such as directed cellular movement and morphological polarization. Here we investigate how an organ-level polarity pattern emerges through symmetry breaking at the cellular level during the formation of a mechanosensory organ. Combining theory, genetic perturbations, and in vivo imaging, we study the development and regeneration of the fluid-motion sensors in the zebrafish's lateral line. We find that two interacting symmetry-breaking events - one mediated by biochemical signaling and the other by cellular mechanics - give rise to precise rotations of cell pairs, which produce a mirror-symmetric polarity pattern in the receptor organ.

Background Optogenetic stimulation has grown into a popular brain stimulation method in basic neuroscience while electrical stimulation predominates in clinical applications. In order to explain the effects of electrical stimulation on a cellular level and evaluate potential advantages of optogenetic therapies, comparisons between the two stimulation modalities are necessary. This comparison is hindered, however, by the difficulty of effectively matching the two fundamentally different modalities. Objective Comparison of brain-wide activation patterns in response to intensity-matched electrical and optogenetic VTA stimulation. Methods We mapped optogenetic and electrical self-stimulation rates in the same mice over stimulation intensity and determined iso-behavioral intensities. Using functional 99mTc-HMPAO SPECT imaging of cerebral blood flow in awake animals, we obtained brain-wide activation patterns for both modalities at these iso-behavioral intensities. We performed these experiments in two mouse lines commonly used for optogenetic VTA stimulation, DAT::Cre and TH::Cre mice. Results We find iso-behavioral intensity matching of stimulation gives rise to similar brain activation patterns. Differences between mouse lines were more pronounced than differences between modalities. Conclusions Previously found large differences of electrical and optogenetic stimulation might be due to unmatched stimulation intensity, particularly relative electrical overstimulation. These findings imply that therapeutic electrical VTA stimulation might be relatively specific if employed with optimized parameters.

Purpose of review To fully exploit the concept of hemodynamic coherence in resuscitating critically ill one should preferably take into account information about the state of parenchymal cells. Monitoring of mitochondrial oxygen tension (mitoPO2) has emerged as a clinical means to assess information of oxygen delivery and oxygen utilization at the mitochondrial level. This review will outline the basics of the technique, summarize its development and describe the rationale of measuring oxygen at the mitochondrial level. Recent findings Mitochondrial oxygen tension can be measured by means of the protoporphyrin IX-Triplet State Lifetime Technique (PpIX-TSLT). After validation and use in preclinical animal models, the technique has recently become commercially available in the form of a clinical measuring system. This system has now been used in a number of healthy volunteer studies and is currently being evaluated in studies in perioperative and intensive care patients in several European university hospitals. Summary PpIX-TSLT is a noninvasive and well tolerated method to assess aspects of mitochondrial function at the bedside. It allows doctors to look beyond the macrocirculation and microcirculation and to take the oxygen balance at the cellular level into account in treatment strategies.

Despite the precise controllability of droplet samples in digital microfluidic (DMF) systems, their capability in isolating single cells for long-time culture is still limited: typically, only a few cells can be captured on an electrode. Although fabricating small-sized hydrophilic micropatches on an electrode aids single-cell capture, the actuation voltage for droplet transportation has to be significantly raised, resulting in a shorter lifetime for the DMF chip and a larger risk of damaging the cells. In this work, a DMF system with 3D microstructures engineered on-chip is proposed to form semi-closed micro-wells for efficient single-cell isolation and long-time culture. Our optimum results showed that approximately 20% of the micro-wells over a 30 × 30 array were occupied by isolated single cells. In addition, low-evaporation-temperature oil and surfactant aided the system in achieving a low droplet actuation voltage of 36V, which was 4 times lower than the typical 150 V, minimizing the potential damage to the cells in the droplets and to the DMF chip. To exemplify the technological advances, drug sensitivity tests were run in our DMF system to investigate the cell response of breast cancer cells (MDA-MB-231) and breast normal cells (MCF-10A) to a widely used chemotherapeutic drug, Cisplatin (Cis). The results on-chip were consistent with those screened in conventional 96-well plates. This novel, simple and robust single-cell trapping method has great potential in biological research at the single cell level. A novel microfluidic device enables researchers to isolate and culture single cells for use in drug testing and other experiments. To date, the high voltages required to capture cells on electrodes of a digital microfluidic (DMF) device have led to a reduced lifespan for the device and potentially damaged the cells. A team led by Yanwei Jia of the University of Macau overcame this by incorporating 3D microstructures into a DMF chip. This constrains the shape of cell culture droplets to isolate and capture single cells. With the use of low evaporation temperature oil and a surfactant, the system enabled them to use one quarter of the voltage of other designs for droplet transportation. The ability to isolate and culture individual cells will be of great value in addressing biological questions at the single-cell level.

At present, early diagnosis and treatment is the most effective way to treat early gastrointestinal neuroendocrine tumors. Therefore, we attempted to carry out multiphoton imaging of early neuroendocrine tumors because of its ability to label-free image tissue microstructure at the cellular level. Imaging results show that this imaging technique can quickly identify the histopathological changes in mucosa and submucosa caused by tumor invasion. Furthermore, we performed automatic image analysis on SHG images and extracted two optical diagnostic features—collagen density and average intensity, and also found obvious differences in the density as well as average intensity of collagen fibers in tumor microenvironment using a series of quantitative analysis. These findings may further facilitate the development of multiphoton microscopic imaging technique for clinical use.

Classical in vivo infection models are oftentimes associated with speculation due to the many physiological factors that are unseen or not accounted for when analyzing experimental outputs, especially when solely utilizing the classic approach of tissue-derived colony-forming unit (CFU) enumeration. To better understand the steps and natural progression of bacterial infection, the pathophysiology of individual organs with which the bacteria interact in their natural course of infection must be considered. In this case, it is not only important to isolate organs as much as possible from additional physiological processes, but to also consider the dynamics of the bacteria at the cellular level within these organs of interest. Here, we describe in detail two models, ex vivo porcine liver and spleen coperfusion and a murine infection model, and the numerous associated experimental outputs produced by these models that can be taken and used together to investigate the pathogen-host interactions within tissues in depth.

Glioblastoma is the most aggressive cancer originating in the brain with an average survival of 15 months. One of the characteristics of glioblastoma is the high level of intra-tumor heterogeneity (ITH), but the composition and complexity at the single-cell level is poorly understood. Here, we aimed to assess the effects and consequences of immune checkpoint inhibitor (ICI) on the cellular and molecular heterogeneity of glioblastoma tumors at the single cell level. In collaboration with the phase I trials unit at Rigshospitalet, we performed paired molecular analysis of glioma cells from primary and relapse surgery after ICI treatment. Samples were analyzed using single-cell RNA sequencing (scRNA-seq) as well as bulk RNA sequencing and whole exome DNA sequencing. In an effort to trace cellular lineages we developed and refined methods to a identify copy number changes using scRNA-seq. To this end, we identified clonal and subclonal tumor cell populations in each sample. We found high levels of ITH prior to treatment, both with respect to the glioblastoma subtype enrichment and the cell type-specific gene expression. Using expression-based cell-type classification, we found defined recurrent cell-type populations present at both surgery time points. The immune checkpoint treatment had consequences on the cellular phenotypes and proportions of tumor cells, suggesting a level of plasticity in the neoplastic cells. Moreover, we identified examples of clonal dynamics and sweeps following ICI treatment, pointing to potential treatment response and resistance in these populations. We additionally found a recurrent pattern of a small population showing high levels of cell cycle activation and simultaneously expressing markers of stem cell potential. In summary, we pursued single cell-focused analysis of ICI treated glioblastoma patients to study the cellular and molecular heterogeneity within and between glioblastoma patients, which pointed to recurrent patterns of cellular responses following ICI treatment.

Background One goal of expression data analysis is to discover the biological significance or function of genes that are differentially expressed. Gene Set Enrichment (GSE) analysis is one of the main tools for function mining that has been widely used. However, every gene expressed in a cell is valuable information for GSE for single-cell RNA sequencing (scRNA-SEQ) data and not should be discarded. Methods We developed the functional expression matrix (FEM) algorithm to utilize the information from all expressed genes. The algorithm converts the gene expression matrix (GEM) into a FEM. The FEM algorithm can provide insight on the biological significance of a single cell. It can also integrate with GEM for downstream analysis. Results We found that FEM performed well with cell clustering and cell-type specific function annotation in three datasets (peripheral blood mononuclear cells, human liver, and human pancreas).

Classic metabolomic methods have proven to be very useful to study functional biology and variation in the chemical composition of different tissues. However, they do not provide any information in terms of spatial localization within fine structures. Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI) does and reaches at best a spatial resolution of 0.25 μm depending on the laser setup, making it a very powerful tool to analyze the local complexity of biological samples at the cellular level. Here, we intend to give an overview of the diversity of the molecules and localizations analyzed using this method as well as to update on the latest adaptations made to circumvent the complexity of samples. MALDI MSI has been widely used in medical sciences and is now developing in research areas as diverse as entomology, microbiology, plant biology, and plant–microbe interactions, the rhizobia symbiosis being the most exhaustively described so far. Those are the fields of interest on which we will focus to demonstrate MALDI MSI strengths in characterizing the spatial distributions of metabolites, lipids, and peptides in relation to biological questions.

The effects of an increase in water temperature as a direct consequence of global change on organisms living in springs and spring brooks have rarely been studied in laboratory experiments. In this study, experiments were conducted to test the response of Gammarus fossarum Koch, 1836, as an abundant representative of the European spring fauna, to changing water temperatures. The aim was to find out experimentally how G. fossarum reacts to varying and increasing water temperatures. The experiments were conducted in flow channels with spring water. In each flow channel, G. fossarum were placed in boxes with a flow-through system for 4 weeks. Two analytical methods were applied: the feeding activity of the amphipods was quantified in order to determine the reaction of G. fossarum on the level of the organism and the respiratory electron transport system (ETS) assay was conducted in order to determine changes in the test organisms on the cellular level. The results show that the feeding activity of G. fossarum increased with increasing water temperature, up to an optimum, indicating an increase in their metabolic activity. The ETS activity does not show significant differences at the different temperatures tested. A possible explanation for this is the ability of the organisms to adapt quickly to the changed environmental circumstances.

Plants are constantly facing abiotic and biotic stresses. To continue to thrive in their environment, they have developed many sophisticated mechanisms to perceive these stresses and provide an appropriate response. There are many ways to study these stress signals in plant, and among them, protoplasts appear to provide a unique experimental system. As plant cells devoid of cell wall, protoplasts allow observations at the individual cell level. They also offer a prime access to the plasma membrane and an original view on the inside of the cell. In this regard, protoplasts are particularly useful to address essential biological questions regarding stress response, such as protein signaling, ion fluxes, ROS production, and plasma membrane dynamics. Here, the tools associated with protoplasts to comprehend plant stress signaling are overviewed and their potential to decipher plant defense mechanisms is discussed.

Recently, rapid advancement in nanomedicine has opened new horizons towards the treatment of disparate types of cancer. Nanomedicine is considered as the science of applying nanoparticles (NPs) for various diagnostic or therapeutic aims. Nanoparticles (NPs) have attracted increasing interest all over the world for the treatment of disparate types of cancer due to their noteworthy properties such as negligible toxicity and great bioactivity. The main objective of this paper is to present a comprehensive review about the potential of various NPs including silver NPs (AgNPs), gold NPs (AuNPs), selenium NPs (SeNPs), titanium oxide NPs (TiO2NPs) and iron oxide NPs (FeONPs) to treat tumoral cells, and investigate the molecular interaction at the cellular level. Moreover, different synthesis mechanisms of NPs along with their operational roles in enhancing the efficiency of conventional chemotherapeutic agents and reducing the toxicity are discussed in detail. Finally, future challenges towards the application of NPs in the field of cancer treatment are presented, and appropriate solutions to remove the ambiguities are suggested.

The tumor immune microenvironment (TIME) encompasses many heterogeneous cell types that engage in extensive crosstalk among the cancer, immune, and stromal components. The spatial organization of these different cell types in TIME could be used as biomarkers for predicting drug responses, prognosis and metastasis. Recently, deep learning approaches have been widely used for digital histopathology images for cancer diagnoses and prognoses. Furthermore, some recent approaches have attempted to integrate spatial and molecular omics data to better characterize the TIME. In this review we focus on machine learning-based digital histopathology image analysis methods for characterizing tumor ecosystem. In this review, we will consider three different scales of histopathological analyses that machine learning can operate within: whole slide image (WSI)-level, region of interest (ROI)-level, and cell-level. We will systematically review the various machine learning methods in these three scales with a focus on cell-level analysis. We will provide a perspective of workflow on generating cell-level training data sets using immunohistochemistry markers to “weakly-label” the cell types. We will describe some common steps in the workflow of preparing the data, as well as some limitations of this approach. Finally, we will discuss future opportunities of integrating molecular omics data with digital histopathology images for characterizing tumor ecosystem.

Photosensitized oxidations, in part responsible for the harmful effect of UV and visible radiation on biological systems, can take place through type I (generation of radicals) and type II (singlet oxygen) mechanisms. Pterins are heterocyclic compounds, widespread in living systems and involved in relevant biological functions. Pterins present a profuse and amazing photochemistry and are endogenous photosensitizers that act mainly via type I mechanism. This survey is aimed to contribute to a better understanding on the complex set of competitive pathways involved in type I photosensitization. Based on studies performed in model systems with pterins as photosensitizers, this review explores the mechanisms involved in the type I photooxidations of proteins, DNA and lipids sensitized by pterins. The generation of radicals and their subsequent reactions are described together with the analysis of the role of O2 and reactive oxygen species. The chemical modification of pterins seeking for better properties and the effect of the photochemical processes at a cellular level are also analyzed.

Simple Summary The fate of the gonad in teleost is influenced by various factors, including genetics and external factors. Several species-specific genes and environmental factors involved in sex determination and differentiation have been identified in teleost. In addition, these factors are species-specific. At the brain level, suppression of key molecule of the hypothalamus–gonadal axis affects sex determination. At the same time, pituitary hormones are required for regulating sex differentiation. However, the role of the brain during sex determination and differentiation remains elusive. In this review, we have gathered and discussed the findings on the role of prominent genes, environmental factors, and the brain in regulating sex determination and differentiation of teleost. Abstract The fish reproductive system is a complex biological system. Nonetheless, reproductive organ development is conserved, which starts with sex determination and then sex differentiation. The sex of a teleost is determined and differentiated from bipotential primordium by genetics, environmental factors, or both. These two processes are species-specific. There are several prominent genes and environmental factors involved during sex determination and differentiation. At the cellular level, most of the sex-determining genes suppress the female pathway. For environmental factors, there are temperature, density, hypoxia, pH, and social interaction. Once the sexual fate is determined, sex differentiation takes over the gonadal developmental process. Environmental factors involve activation and suppression of various male and female pathways depending on the sexual fate. Alongside these factors, the role of the brain during sex determination and differentiation remains elusive. Nonetheless, GnRH III knockout has promoted a male sex-biased population, which shows brain involvement during sex determination. During sex differentiation, LH and FSH might not affect the gonadal differentiation, but are required for regulating sex differentiation. This review discusses the role of prominent genes, environmental factors, and the brain in sex determination and differentiation across a few teleost species.

Simple Summary Magnetic hyperthermia therapy is an alternative treatment for cancer that complements traditional therapies and that has shown great promise in recent years. In this review, we assess the current applications of this therapy in order to understand why its translation from the laboratory to the clinic has been less smooth than was anticipated, identifying the possible bottlenecks and proposing solutions to the problems encountered. Abstract Hyperthermia has emerged as a promising alternative to conventional cancer therapies and in fact, traditional hyperthermia is now commonly used in combination with chemotherapy or surgery during cancer treatment. Nevertheless, non-specific application of hyperthermia generates various undesirable side-effects, such that nano-magnetic hyperthermia has arisen a possible solution to this problem. This technique to induce hyperthermia is based on the intrinsic capacity of magnetic nanoparticles to accumulate in a given target area and to respond to alternating magnetic fields (AMFs) by releasing heat, based on different principles of physics. Unfortunately, the clinical implementation of nano-magnetic hyperthermia has not been fluid and few clinical trials have been carried out. In this review, we want to demonstrate the need for more systematic and basic research in this area, as many of the sub-cellular and molecular mechanisms associated with this approach remain unclear. As such, we shall consider here the biological effects that occur and why this theoretically well-designed nano-system fails in physiological conditions. Moreover, we will offer some guidelines that may help establish successful strategies through the rational design of magnetic nanoparticles for magnetic hyperthermia.

We performed a histological and quantitative study of iron in archaeological maize seeds from prehispanic times recovered from Tarapacá, Atacama Desert. Also, we examined iron distribution changes at the cell level in embryos from ancient versus new varieties of maize. Our results show a progressive decrease in iron concentration from the oldest maize to modern specimens. We interpret the results as an effect of prehispanic agriculture over the micronutrient composition of maize.

International audience; Background and PurposeDopamine in the striatum plays a crucial role in reward processes and action selection. Dopamine signals are transduced by D1 and D2 dopamine receptors which trigger mirror effects through the cAMP/PKA signalling cascade in D1 and D2 medium-sized spiny neurones (MSNs). Phosphodiesterases (PDEs), which determine the profile of cAMP signals, are highly expressed in MSNs, but their respective roles in dopamine signal integration remain poorly understood.Experimental approachWe used genetically-encoded FRET biosensors to monitor at the single cell level the functional contribution of PDE2A, PDE4 and PDE10A in the changes of the cAMP/PKA response to transient and continuous dopamine in mouse striatal brain slices.Key ResultsWe found that PDE2A, PDE4 and PDE10A operate on the moderate to high cAMP levels elicited by D1 or A2A receptor stimulation. In contrast, only PDE10A is able to reduce cAMP down to baseline in both type of neurones, leading to the dephosphorylation of PKA substrates.Conclusion and ImplicationsIn both MSN types, PDE10A inhibition blunts the responsiveness to dopamine, whereas PDE2A or PDE4 inhibition reinforces dopamine action.

In combination with scRNA-seq, spatial transcriptomics have the potential to lead to an unprecedented view of lung architecture at the single cell level, providing original information on lung physiology and physiopathologyhttps://bit.ly/3uDYir0

Throughout life, the body is subjected to various mechanical forces on the organ, tissue, and cellular level. Mechanical stimuli are essential for organ development and function. One organ whose function depends on the tightly connected interplay between mechanical cell properties, biochemical signaling, and external forces is the lung. However, altered mechanical properties or excessive mechanical forces can also drive the onset and progression of severe pulmonary diseases. Characterizing the mechanical properties and forces that affect cell and tissue function is therefore necessary for understanding physiological and pathophysiological mechanisms. In recent years, multiple methods have been developed for cellular force measurements at multiple length scales, from subcellular forces to measuring the collective behavior of heterogeneous cellular networks. In this short review, we give a brief overview of the mechanical forces at play on the cellular level in the lung. We then focus on the technological aspects of measuring cellular forces at many length scales. We describe tools with a subcellular resolution and elaborate measurement techniques for collective multicellular units. Many of the technologies described are by no means restricted to lung research and have already been applied successfully to cells from various other tissues. However, integrating the knowledge gained from these multi-scale measurements in a unifying framework is still a major future challenge.

Single-cell RNA sequencing (scRNA-seq) technology can analyze the transcriptome expression level of cells with high-throughput from the single cell level, fully show the heterogeneity of cells, and provide a new way for the study of multicellular biological heterogeneity. Synovitis is the pathological basis of rheumatoid arthritis (RA). Synovial fibroblasts (SFs) and synovial macrophages are the core target cells of RA, which results in the destruction of articular cartilage, as well as bone. Recent scRNA-seq technology has made breakthroughs in the differentiation and development of two types of synovial cells, identification of subsets, functional analysis, and new therapeutic targets, which will bring remarkable changes in RA treatment.

The current ecosystem of single-cell RNA-seq platforms is rapidly expanding, but robust solutions for single-cell and single-molecule full-length RNA sequencing are virtually absent. A high-throughput solution that covers all aspects is necessary to study the complex life of mRNA on the single-cell level. The Nanopore platform offers long read sequencing and can be integrated with the popular single-cell sequencing method on the 10× Chromium platform. However, the high error-rate of Nanopore reads poses a challenge in downstream processing (e.g., for cell barcode assignment). We propose a solution to this particular problem by using a hybrid sequencing approach on Nanopore and Illumina platforms. Our software ScNapBar enables cell barcode assignment with high accuracy, especially if sequencing saturation is low. ScNapBar uses unique molecular identifier (UMI) or Naïve Bayes probabilistic approaches in the barcode assignment, depending on the available Illumina sequencing depth. We have benchmarked the two approaches on simulated and real Nanopore data sets. We further applied ScNapBar to pools of cells with an active or a silenced nonsense-mediated RNA decay pathway. Our Nanopore read assignment distinguishes the respective cell populations and reveals characteristic nonsense-mediated mRNA decay events depending on cell status.

Species that can regrow their lost appendages have been studied with the ultimate aim of developing methods to enable human limb regeneration. These examinations highlight that appendage regeneration progresses through shared tissue stages and gene activities, leading to the assumption that appendage regeneration paradigms (e.g. tails and limbs) are the same or similar. However, recent research suggests these paradigms operate differently at the cellular level, despite sharing tissue descriptions and gene expressions. Here, collecting the findings from disparate studies, I argue appendage regeneration is context dependent at the cellular level; nonetheless, it requires (i) signalling centres, (ii) stem/progenitor cell types and (iii) a regeneration-permissive environment, and these three common cellular principles could be more suitable for cross-species/paradigm/age comparisons.

Organoids have emerged as powerful model systems to study organ development and regeneration at the cellular level. Recently developed microscopy techniques that track individual cells through space and time hold great promise to elucidate the organizational principles of organs and organoids. Applied extensively in the past decade to embryo development and 2D cell cultures, cell tracking can reveal the cellular lineage trees, proliferation rates, and their spatial distributions, while fluorescent markers indicate differentiation events and other cellular processes. Here, we review a number of recent studies that exemplify the power of this approach, and illustrate its potential to organoid research. We will discuss promising future routes, and the key technical challenges that need to be overcome to apply cell tracking techniques to organoid biology.

Publisher Copyright: © 2020 The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. The heartwood of many wood species is rich in extractives, which improve the wood material's resistance to biological attack. Their concentration is generally higher in outer than inner heartwood, but the exact radial changes in aging heartwood remain poorly characterized. This investigation studied these radial changes in detail in Scots pine (Pinus sylvestris L.), using radial sample sequences prepared from three different trees. Stilbene and resin acid contents were first measured from bulk samples, after which the extractive contents of individual heartwood annual rings were investigated using Raman spectroscopy and fluorescence microscopy. Raman imaging and fluorescence microscopy were also used to study the cellular-level distributions of extractives in different annual rings. Although there were substantial differences between the trees, the content and distribution of stilbenes seemed to follow a general radial trend. The results suggest that stilbenes are absorbed into heartwood tracheid cell walls from small stilbene-rich extractive deposits over several years and then eventually transform into non-extractable compounds in aging heartwood. Resin acids followed no consistent radial trends, but their content was strongly connected to the frequency of large extractive deposits in latewood tracheid lumens. The results highlight the variability of heartwood extractives: their content and distribution vary not only between trees but also between and even within the annual rings of a single tree. This high variability is likely to have important effects on the properties of heartwood and the utilization of heartwood timber.

The International Comet Assay Workshops (ICAW) are a series of scientific conferences dealing with different aspects of the comet assay. The assay itself is a simple method for the detection of DNA strand breaks at the cellular level and can be applied to any cell type derived from different organs and tissues of eukaryotic organisms. Additionally, the comet assay is widely applied in human biomonitoring, ecotoxicology, genotoxicity testing of chemicals, but also in basic research studying the mechanisms of DNA damage and repair. The 2019 ICAW edition gathered about 80 participants with over 30 lecturers, 27 poster presentations and 2 open discussion sessions presenting the latest advances in technical developments as well as applications of the comet assay in genetic toxicology, and environmental and human biomonitoring. This report summarises the important issues that were raised and discussed during the sessions as well as a short synopsis of the papers selected for inclusion in this special issue. Based on the topics presented at the workshop, the assay with its new modifications and applications has a bright future and will for sure stay one of the most popular methods in genetic toxicology and beyond in the years to come.

Several experimental and clinical studies have transformed the traditional antimalarial role of chloroquine (CHQ) and related structural analogues to potent therapeutic agents for a host of nonmalarial indications. The expanding clinical applicability for these drugs includes rheumatological and cardiovascular disorders (CVD), chronic kidney disease (CKD), oncology, and a variety of nonmalarial infections. These clinical advancements are primarily related to pleiotropic pharmacological actions of these drugs, including immunomodulation, anti-inflammatory properties, and capabilities of inducing autophagy and apoptosis at a cellular level. Historically, many clinical benefits in nonmalarial indications were first recognized through serendipitous observations; however, with numerous ongoing systematic clinical studies, the clinical horizons of these drugs have a promising future.