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OBJECTIVEFailure to close the ductus arteriosus immediately post-birth, patent ductus arteriosus (PDA), accounts for up to 10% of all congenital heart defects. Despite significant advances in PDA management options, including pharmacological treatment targeting the prostaglandin pathway, a proportion of patients fail to respond and must undergo surgical intervention. Thus, further refinement of the cellular and molecular mechanisms that govern vascular remodeling of this vessel is required.APPROACH AND RESULTSAs anticipated, single-cell RNA sequencing on the ductus arteriosus in mouse embryos at E18.5, P0.5, and P5, revealed broad transcriptional alterations in the endothelial, smooth muscle, and fibroblast cell compartments. Making use of these data sets, vimentin emerged as an interesting candidate for further investigation. Subsequent studies demonstrated that, in fact, mice with genetic deletion of vimentin fail to complete vascular remodeling of the ductus arteriosus, as per presence of a functional lumen.CONCLUSIONSThrough single-cell RNA-sequencing and by tracking closure of the ductus arteriosus postnatally in mice, we uncovered the unexpected contribution of vimentin in driving complete closure of the ductus arteriosus potentially through regulation of the Notch signaling pathway.HIGHLIGHTSSingle-cell RNA-sequencing on the ductus arteriosus at E18.5, P0.5, and P5 reveals how the ductus arteriosus undergoes drastic transcriptional changes at the single-cell level.Endothelial cells increase levels of Vimentin, Notch1 and Jag1 transcripts soon after birth (P0.5), concurrent with ductus arteriosus closure.Loss of vimentin, the major intermediate filament protein of endothelial cells, prevents proper permanent closure of the ductus arteriosus.

Extracellular vesicles (EVs) are membrane vesicles secreted by cells, playing critical roles in mediating intercellular communications for various physiological and pathological processes. Most of the EV analysis is currently performed at the bulk level, obscuring the origin of the EVs and diverse characteristics of the individual extracellular vesicle. Technologies to analyze the extracellular vesicles at the single-cell and single-vesicle levels are needed to evaluate EV comprehensively and decode the heterogeneity underlying EV secretion. Microfluidic platforms that could control and manipulate fluids at the microscale provide an efficient way to achieve the aims. Various microfluidics-based technologies are emerging to realize single-cell EV secretion analysis and single EV analysis, which would be summarized in this mini-review.

Long non-coding RNAs (LncRNAs) are a type of RNA with little or no protein-coding ability. Their length is more than 200 nucleotides. A large number of studies have indicated that lncRNAs play a significant role in various biological processes, including chromatin organizations, epigenetic programmings, transcriptional regulations, post-transcriptional processing, and circadian mechanism at the cellular level. Since lncRNAs perform vast functions through their interactions with proteins, identifying lncRNA-protein interaction is crucial to the understandings of the lncRNA molecular functions. However, due to the high cost and time-consuming disadvantage of experimental methods, a variety of computational methods have emerged. Recently, many effective and novel machine learning methods have been developed. In general, these methods fall into two categories: semisupervised learning methods and supervised learning methods. The latter category can be further classified into the deep learning-based method, the ensemble learning-based method, and the hybrid method. In this paper, we focused on supervised learning methods. We summarized the state-of-the-art methods in predicting lncRNA-protein interactions. Furthermore, the performance and the characteristics of different methods have also been compared in this work. Considering the limits of the existing models, we analyzed the problems and discussed future research potentials.

Bone is composed of dense and solid cortical bone and honeycomb-like trabecular bone. Although cortical bone provides the majority of mechanical strength for a bone, there are few studies focusing on cortical bone repair or regeneration. Osteons (the Haversian system) form structural and functional units of cortical bone. In recent years, emerging evidences have shown that the osteon structure (including osteocytes, lamellae, lacunocanalicular network, and Haversian canals) plays critical roles in bone mechanics and turnover. Therefore, reconstruction of the osteon structure is crucial for cortical bone regeneration. This article provides a systematic summary of recent advances in osteons, including the structure, function, turnover, and regenerative strategies. First, the hierarchical structure of osteons is illustrated and the critical functions of osteons in bone dynamics are introduced. Next, the modeling and remodeling processes of osteons at a cellular level and the turnover of osteons in response to mechanical loading and aging are emphasized. Furthermore, several bioengineering approaches that were recently developed to recapitulate the osteon structure are highlighted. IMPACT STATEMENT: This review provides a comprehensive summary of recent advances in osteons, especially the roles in bone formation, remodeling, and regeneration. Besides introducing the hierarchical structure and critical functions of osteons, we elucidate the modeling and remodeling of osteons at a cellular level. Specifically, we highlight the bioengineering approaches that were recently developed to mimic the hierarchical structure of osteons. We expect that this review will provide informative insights and attract increasing attentions in orthopedic community, shedding light on cortical bone regeneration in the future.

Liver metastasis, the leading cause of colorectal cancer mortality, exhibits a highly heterogeneous and suppressive immune microenvironment. Here, we sequenced 97 matched samples by using single-cell RNA sequencing and spatial transcriptomics. Strikingly, the metastatic microenvironment underwent remarkable spatial reprogramming of immunosuppressive cells such as MRC1+ CCL18+ M2-like macrophages. We further developed scMetabolism, a computational pipeline for quantifying single-cell metabolism, and observed that those macrophages harbored enhanced metabolic activity. Interestingly, neoadjuvant chemotherapy could block this status and restore the antitumor immune balance in responsive patients, whereas the nonresponsive patients deteriorated into a more suppressive one. Our work described the immune evolution of metastasis and uncovered the black box of how tumors respond to neoadjuvant chemotherapy. Significance: We present a single-cell and spatial atlas of colorectal liver metastasis and found the highly metabolically activated MRC1+ CCL18+ M2-like macrophages in metastatic sites. Efficient neoadjuvant chemotherapy can slow down such metabolic activation, raising the possibility to target metabolism pathways in metastasis. This article is highlighted in the In This Issue feature, p. 1

We report how Raman difference imaging provides insight on cellular biochemistry in vivo as a function of sub-cellular dimensions and the cellular environment. We show that this approach offers a sensitive diagnostic to address blood biochemistry at the cellular level. We examine Raman microscopic images of the distribution of the different hemoglobins in both healthy (discocyte) and unhealthy (echinocyte) blood cells and interpret these images using pre-calculated, accurate pre-resonant Raman tensors for scattering intensities specific to hemoglobins. These tensors are developed from theoretical calculations of models of the oxy, deoxy and met forms of heme benchmarked against the experimental visible spectra of the corresponding hemoglobins. The calculations also enable assignments of the electronic transitions responsible for the colour of blood: these are mainly ligand to metal charge transfer transitions., We assign the electronic transitions responsible for the colour of blood and present a Raman imaging diagnostic approach for individual blood cells.

Recent emergence of FTIR spectromicroscopy (micro-FTIR) as a dynamic spectroscopy for imaging to the study of biological chemistry has opened new possibilities for investigating in situ drug release, redox chemistry effects with biological molecules, DNA and drug interactions, membranes dynamic, and redox reaction with proteins at the single cell level. Micro-FTIR applied to metallodrugs have been playing important role in the last decade because of its great potential to achieve more robust and controlled pharmacological effects against several diseases, including cancer. An important aspect in the development of these drugs is to understand their cellular properties, such as uptake, accumulation, activity, and toxicity. In this review, we present the potential application of the micro-FTIR and its importance for studying metal-based drugs, highlighting the prospective of chemistry of living cells. We also present a bioimaging emphasis, which is therefore of high importance to localize the cellular processes, for the proper understanding of the mechanism of action.

Background: Hypertrophic cardiomyopathy (HCM) is a complex disease partly explained by the effects of individual gene variants on sarcomeric protein biomechanics. At the cellular level, HCM mutations most commonly enhance force production, leading to higher energy demands. Despite significant advances in elucidating sarcomeric structure–function relationships, there is still much to be learned about the mechanisms that link altered cardiac energetics to HCM phenotypes. In this work, we test the hypothesis that changes in cardiac energetics represent a common pathophysiologic pathway in HCM. Methods: We performed a comprehensive multiomics profile of the molecular (transcripts, metabolites, and complex lipids), ultrastructural, and functional components of HCM energetics using myocardial samples from 27 HCM patients and 13 normal controls (donor hearts). Results: Integrated omics analysis revealed alterations in a wide array of biochemical pathways with major dysregulation in fatty acid metabolism, reduction of acylcarnitines, and accumulation of free fatty acids. HCM hearts showed evidence of global energetic decompensation manifested by a decrease in high energy phosphate metabolites (ATP, ADP, and phosphocreatine) and a reduction in mitochondrial genes involved in creatine kinase and ATP synthesis. Accompanying these metabolic derangements, electron microscopy showed an increased fraction of severely damaged mitochondria with reduced cristae density, coinciding with reduced citrate synthase activity and mitochondrial oxidative respiration. These mitochondrial abnormalities were associated with elevated reactive oxygen species and reduced antioxidant defenses. However, despite significant mitochondrial injury, HCM hearts failed to upregulate mitophagic clearance. Conclusions: Overall, our findings suggest that perturbed metabolic signaling and mitochondrial dysfunction are common pathogenic mechanisms in patients with HCM. These results highlight potential new drug targets for attenuation of the clinical disease through improving metabolic function and reducing mitochondrial injury.

Could enzymatic activities and their cooperative functions act as cellular temperature-sensing systems? This review introduces recent opto-thermal technologies for microscopic analyses of various types of cellular temperature-sensing system. Optical microheating technologies have been developed for local and rapid temperature manipulations at the cellular level. Advanced luminescent thermometers visualize the dynamics of cellular local temperature in space and time during microheating. An optical heater and thermometer can be combined into one smart nanomaterial that demonstrates hybrid function. These technologies have revealed a variety of cellular responses to spatial and temporal changes in temperature. Spatial temperature gradients cause asymmetric deformations during mitosis and neurite outgrowth. Rapid changes in temperature causes imbalance of intracellular Ca2+ homeostasis and membrane potential. Among those responses, heat-induced muscle contractions are highlighted. It is also demonstrated that the short-term heating hyperactivates molecular motors to exceed their maximal activities at optimal temperatures. We discuss future prospects for opto-thermal manipulation of cellular functions and contributions to obtain a deeper understanding of the mechanisms of cellular temperature-sensing systems.

Understanding cellular metabolism (how cells use energy) can be key in treating a wide range of diseases, including vascular disease and cancer. Although many techniques can measure these processes in tens of thousands of cells, researchers have not been able to measure them at the single-cell level. Researchers have used a genetically encoded biosensor with artificial intelligence to measure glycolysis. (Process of converting glucose to energy, single endothelial cells, blood vessel cells). Keywords:Cancer; Cells; Tissues, Tumors; Prevention, Prognosis; Diagnosis; Imaging; Screening; Treatment; Management https://www.raftpubs.com/jca-chemistry/articles/jca_raft1016.php

Local anesthetic systemic toxicity (LAST) is a life-threatening event caused by elevated local anesthetic plasma concentration. It is often unrecognized or misdiagnosed. Peripartum women are at increased risk for toxicity due to pregnancy-related physiological changes. Rising serum drug levels can cause cellular level impairment of mitochondria and voltage-gated ion channels leading to a cascade of symptoms that can end in cardiac arrest. Local anesthetic systemic toxicity can mimic other maternal pathologies but may be considered if local anesthetics have been used. Published treatment guidelines for this event include lipid emulsion which is approved for use in pregnant women. We review LAST in the maternity care setting, published treatment protocols, management of maternity patients with toxicity, and recommendations to increase awareness among maternity care clinicians for this medical emergency.

It has been nearly 15 years since the discovery of human-induced pluripotent stem cells (iPSCs). During this time, differentiation methods to targeted cells have dramatically improved, and many types of cells in the human body can be currently generated at high efficiency. In the cardiovascular field, the ability to generate human cardiomyocytes in vitro with the same genetic background as patients has provided a great opportunity to investigate human cardiovascular diseases at the cellular level to clarify the molecular mechanisms underlying the diseases and discover potential therapeutics. Additionally, iPSC-derived cardiomyocytes have provided a powerful platform to study drug-induced cardiotoxicity and identify patients at high risk for the cardiotoxicity; thus, accelerating personalized precision medicine. Moreover, iPSC-derived cardiomyocytes can be sources for cardiac cell therapy. Here, we review these achievements and discuss potential improvements for the future application of iPSC technology in cardiovascular diseases.

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.

In this paper, we take a look at Austria’s renewable energy targets established in the Renewable Energy Expansion Act (EAG), aiming to annually generate an additional 10 TWh of wind power by 2030. We conduct a GIS (geographic information system)-based analysis to determine average wind power density in Austria on a cellular level while considering prohibited regions, such as national parks, where building wind turbines might not be allowed. The calculated expansion potential for all remaining regions of Austria is allocated to the closest corresponding transmission nodes. Furthermore, we suggest an optimization algorithm to geographically distribute the expansion of wind power capacity to applicable transmission nodes. Finally, we conduct a case study to validate the algorithm using historical data on expansion and utilize it to predict an annual scenario for wind power expansion from 2021 to 2030 on a regional level. The total expansion required to achieve the goal of 10 TWh is assessed to be 4 GW based on predefined full load hours while assuming an exponential increase in annually added capacity (from 250 MW in 2021 to 590 MW in 2030).

Background: In recent years much research has been devoted to the deployment of biomarkers in the field of heart failure. Objectives: To study the potential of post-transcriptional regulation by microRNAs on the diagnosis, management and therapy of heart failure. Methods: Literature search focus on the role of microRNAs in heart failure. Results: MicroRNAs are expressed and regulated in the course of the pathological manifestations of heart failure (HF). This wide and uncharted area of genetic imprints consisting of small non-coding RNA molecule is upregulated and released into the bloodstream from organs under certain conditions and or stress. The use of genetically based strategies for the management of HF has gained great interest in the field of biomedical science because they can be used as biomarkers providing information regarding cardiac status and function. They also appear as promising tools with therapeutic potential because of their ability to induce changes at the cellular level without creating alterations in the gene sequence. In addition, with the advances in genomic sequencing, quantification and synthesis in technologies of microRNAs identification as well as the growing knowledge of the biology of miRNAs and their involvement in HF, it is expected to favorably affect the prognosis of HF patients. Conclusion: MicroRNAs are involved in the regulation of multibiological processes involved in the progress of heart failure. More studies are needed to achieve a clinical valuable implementation of microRNAs in the management of HF.

Introduction. Liver diseases with all the variety of clinical manifestations have common pathogenetic links at the cellular level. The group of hepatoprotective agents is represented by drugs that exhibit versatile mechanisms for protecting liver cells from the effects of damaging factors, the main of which are membrane-stabilizing, antioxidant, regenerative, detoxifying, choleretic and anti-inflammatory effects. The high therapeutic and hepatoprotective effectiveness of modern drugs is largely due to their metabolic effects, as well as their ability to bind free radicals and reactive oxygen species in the cell.Text. The purpose of this work is to form an analytical review of the literature on the assortment and concepts of improving the dosage forms of hepatoprotective agents. According to the analysis and systematization of modern publications devoted to the use of hepatoprotectors in the treatment of liver diseases, it is shown that it is necessary to develop new formulations and combinations of biologically active substances with the manifestation of versatile mechanisms of hepatoprotection, as well as to improve the composition and manufacturing technologies of existing traditional therapies. One of the current trends is the use of new substances in the development of traditional and innovative dosage forms. The search for biologically active molecules with antioxidant, antiradical and membrane-stabilizing activity that can be considered as effective hepatoprotectors continues. An integral task of pharmaceutical development is the creation of bioavailable drugs that have a prolonged effect and minimal side effects. A promising direction in pharmaceutical technology is the development of innovative drugs for the directed transport of biologically active molecules to the affected organ.Conclusion. As a result of the analysis of modern data, priority directions for the development and improvement of existing formulations based on modern approaches to the production of innovative dosage forms are identified. The relevance of improving the dosage forms of hepatoprotectors presented on the pharmaceutical market is shown. Of particular interest is the development of innovative targeted delivery systems with effective and safe hepatoprotectors in various combinations, including those based on cinnamic acid derivatives.

The cardiac pacemaker ignites and coordinates the contraction of the whole heart, uninterruptedly, throughout our entire life. Pacemaker rate is constantly tuned by the autonomous nervous system to maintain body homeostasis. Sympathetic and parasympathetic terminals act over the pacemaker cells as the accelerator and the brake pedals, increasing or reducing the firing rate of pacemaker cells to match physiological demands. Despite the remarkable reliability of this tissue, the pacemaker is not exempt from the detrimental effects of aging. Mammals experience a natural and continuous decrease in the pacemaker rate throughout the entire lifespan. Why the pacemaker rhythm slows with age is poorly understood. Neural control of the pacemaker is remodeled from birth to adulthood, with strong evidence of age-related dysfunction that leads to a downshift of the pacemaker. Such evidence includes remodeling of pacemaker tissue architecture, alterations in the innervation, changes in the sympathetic acceleration and the parasympathetic deceleration, and alterations in the responsiveness of pacemaker cells to adrenergic and cholinergic modulation. In this review, we revisit the main evidence on the neural control of the pacemaker at the tissue and cellular level and the effects of aging on shaping this neural control.

Adaptation to mechanical loading is critical to maintaining bone mass and offers therapeutic potential to preventing age-related bone loss and osteoporosis. However, increasing the duration of loading is met with "diminishing returns" as the anabolic response quickly becomes saturated. As a result, the anabolic response to daily activities and repetitive bouts of loading is limited by the underlying mechanisms that desensitize and render bone unresponsive at the cellular level. Osteocytes are the primary cells that respond to skeletal loading and facilitate the overall anabolic response. Although many of osteocytes' signaling mechanisms activated in response to loading are considered anabolic in nature, several of them can also render osteocytes insensitive to further stimuli and thereby creating a negative feedback loop that limits osteocytes' overall response. The purpose of this review is to examine the potential mechanisms that may contribute to the loss of mechanosensitivity. In particular, we examined the inactivation/desensitization of ion channels and signaling molecules along with the potential role of endocytosis and cytoskeletal reorganization. The significance in defining the negative feedback loop is the potential to identify unique targets for enabling osteocytes to maintain their sensitivity. In doing so, we can begin to cultivate new strategies that capitalize on the anabolic nature of daily activities that repeatedly load the skeleton.

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.

Non-viral vehicles hold therapeutic promise in advancing the delivery of a variety of cargos in vitro and in vivo, including small molecule drugs, biologics, and especially nucleic acids. However, their efficacy at the cellular level is limited by several delivery barriers, with endolysosomal degradation being most significant. The entrapment of vehicles and their cargo in the acidified endosome prevents access to the cytosol, nucleus, and other subcellular compartments. Understanding the factors that contribute to uptake and intracellular trafficking, especially endosomal entrapment and release, is key to overcoming delivery obstacles within cells. In this review, we summarize and compare experimental techniques for assessing the extent of endosomal escape of a variety of non-viral vehicles and describe proposed escape mechanisms for different classes of lipid-, polymer-, and peptide-based delivery agents. Based on this evaluation, we present forward-looking strategies utilizing information gained from mechanistic studies to inform the rational design of efficient delivery vehicles.

Cardiac arrhythmias are prevalent among humans across all age ranges, affecting millions of people worldwide. While cardiac arrhythmias vary widely in their clinical presentation, they possess shared complex electrophysiologic properties at cellular level that have not been fully studied. Over the last decade, our current understanding of the functional roles of non-coding RNAs have progressively increased. microRNAs represent the most studied type of small ncRNAs and it has been demonstrated that miRNAs play essential roles in multiple biological contexts, including normal development and diseases. In this review, we provide a comprehensive analysis of the functional contribution of non-coding RNAs, primarily microRNAs, to the normal configuration of the cardiac action potential, as well as their association to distinct types of arrhythmogenic cardiac diseases.

DNA computation is considered as one fascinating alternative to silicon-based computers, which has evoked substantial attention and gained rapid advancements. Besides realizing versatile functions, implementing spatiotemporal control of logic operations, especially in cellular level, is also of great significance to the development of DNA computation. However, developing simple and efficient methods to restrict DNA logic gates performing in live cells is still a challenge. In this work, we designed a series of DNA logic gates by taking full advantage of the diversity and programmability of G-quadruplex (G4) structure. More importantly, by further utilizing the high affinity and specific endocytosis of cells to aptamer G4, we have achieved in realizing an INHIBIT logic gate whose operational site is precisely restricted in specific live cells. The design strategy may have great potential in the field of molecular computation and smart bio-applications.

The paper analyzes methodological foundations for selection of educational content about the structure and life of protozoa in the university course Zoology while training bachelors of Pedagogical education majoring in Biology and Chemistry. The authors test the hypothesis that the methodological foundations of the study of protozoa are the achievements of biological sciences, as well as formulated methodological constructs based on the recognition of ecological and evolutionary approaches. General scientific methods of theoretical knowledge as well as methods used at the empirical and theoretical levels of knowledge were used as research methods. The methodological foundations for selecting the educational content about the structure and life of protozoa in the university course Zoology are: general properties of living organisms, levels of life organization, ideas of development and structure, an ecological-evolutionary approach, description of co-evolutionary processes and the role of cognitive models. The belonging of unicellular organisms to the cellular level of life organization does not prevent their description taking into account the structural-functional approach with the substantiation of a close connection between the vital processes of unicellular and multicellular organisms. Methodological attitudes operating in biological knowledge are associated with an orientation towards the idea of development and organization. The study of unicellular organisms biology taking into account the above ideas is guided by ecological and evolutionary aspects. The formulated approach is important from a practical and theoretical point of view. The aspects of the practical application of knowledge about the biology of protozoa include, for example, the study of their role in the life of the biosphere. Evolutionary aspects in the description of protozoa are realized at the phylogenetic and ontogenetic levels.

Single cell analysis has drawn increasing interest from the research community due to its capability to interrogate cellular heterogeneity, allowing refined tissue classification and facilitating novel biomarker discovery. With the advancement of relevant instruments and techniques, it is now possible to perform multiple omics including genomics, transcriptomics, metabolomics or even proteomics at single cell level. In comparison with other omics studies, single-cell metabolomics (SCM) represents a significant challenge since it involves many types of dynamically changing compounds with a wide range of concentrations. In addition, metabolites cannot be amplified. Although difficult, considerable progress has been made over the past decade in mass spectrometry (MS)-based SCM in terms of processing technologies and biochemical applications. In this review, we will summarize recent progress in the development of promising MS platforms, sample preparation methods and SCM analysis of various cell types (including plant cell, cancer cell, neuron, embryo cell, and yeast cell). Current limitations and future research directions in the field of SCM will also be discussed.

Exposure to outdoor conditions can cause chemical components on wood surfaces to deteriorate. To develop effective wood protection methods, a deep understanding of the mechanisms of weathering-induced wood surface degradation is required. However, the effects of outdoor exposure on wood at the cellular level have not been investigated in detail. Additionally, a comparative study of the degradation behavior of sapwood and heartwood has also not been conducted. This paper investigated the chemical changes in cell walls in Japanese cedar (Cryptomeria japonica D. Don) sapwood and heartwood during natural weathering using confocal Raman microscopy. Spectral and chemical mapping revealed that heartwood had higher weather durability than sapwood due to the large extractives content in heartwood. Although there were differences in the rates of lignin reduction between sapwood and heartwood, the molecular structures and lignin-degraded sites of the weathered samples were almost identical. The degradation patterns of naturally weathered wood were also similar to that of artificially weathered wood. The knowledge revealed here will help in the development of more effective wood protection methods.

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.

Since wood chemical components can be depolymerized and modified by weathering, a better understanding of the mechanisms governing these processes in needed to develop effective protection methods for wood surfaces. Unfortunately, very little has been reported about the micro-scale chemical changes in wood, particularly hardwood, during weathering. The purpose of the present work is to determine the degradation behavior of Japanese beech (Fagus crenata Blume) under artificial weathering at the cellular-level. Herein, the structural and micro-distributional changes in wood components during weathering were investigated using micro-Raman spectral and chemical mapping analyses. The Raman spectra showed that weathering facilitated lignin degradation and modification. The degradation behavior of lignin differed depending on the type of wood tissue. The rate of lignin reduction followed a descending order: vessel element > axial parenchyma cell > wood fiber. Raman mapping determined that cellular-level lignin reduction on the surface layers differed for wood species. Although lignin degradation of cedar tracheids proceeded from both the surface and the cell lumen, the lignin in beech fibers degraded according to the depth.

Cytoplasmic microviscosity (CPMV) plays essential roles in governing the diffusion-mediated cellular processes and has been recognized as a reliable indicator of the cellular response of many diseases and malfunctions. Current CPMV studies are exclusively established by probe-assisted optical methods, which nevertheless necessitate the complicated synthesis and delivery of optical probes into cells and thus the issues of biocompatibility and bio-orthogonality. Using twin nanopipettes integrated with a patch-clamp system, a practical electrochemical single-cell measurement is presented, which is capable of real-time and long-term CPMV detection without cell disruption. Specifically, upon the operation of the twin nanopipettes, the cellular CPMV status, which is correlated to cytoplasmic ionic mobility, could be sensibly transduced via the ionic current passing through the nanosystem. The average CPMV value of HeLa cells was detected as ca. 86 cP. Notably, the correlation between chemotherapy and CPMV alterations makes this approach possible for the real-time and long-term assessment of the evolution of external stimuli, as exemplified by the two natural products taxol and colchicine. Integrated with the patch-clamp setup, this study features the first use of twin nanopipettes for electrochemical CPMV monitoring of single living cells, and it is expected to inspire more interest in the exploitation of dual- and multiple nanopipettes for advanced single-cell analysis.

Aim. To analyze the factors associated with the development of chronic heart failure (CHF) in older age and to evaluate possible approaches to managing outpatients.Material and methods. The search for domestic and foreign publications in Russian and international systems (PubMed, eLibrary, Medscape, etc.) for the last 0.5–15 years has been carried out. The analysis includes articles from the peer-reviewed literature.Results. During aging, a number of structural changes and changes at the cellular level occur in the cardiovascular system, predisposing to the development of myocardial dysfunction, which explains the variety of manifestations of heart failure and the prevalence of preserved left ventricular ejection fraction (LVEF). Involutional functional and morphological changes in organs and systems form multimorbidity and nonspecificity of clinical symptoms and signs. «Cross symptoms» of frailty and CHF often complicate the timely diagnosis of the heart failure. Medications used for the treatment of CHF in older and younger patients are similar, but when choosing a specific drug, caution should be exercised in the group of the most vulnerable patients: over the age of 85, in the first two weeks after discharge from the hospital and in the presence of frailty.Conclusion. Based on the available results of studies, it is necessary to be alert of primary care physicians regarding the presence of CHF with preserved LVEF in older persons with multimorbidity and geriatric syndromes. The therapeutic strategy for CHF older patients is complex and involves an individualized approach, depending on the clinical situation.

Coronaviruses are a family of viruses present in various animals, and alpha and beta types can infect humans. Human coronavirus varieties settle in different regions of the respiratory tract epithelium, causing disease with varying severity. We conducted a comprehensive academic search to aggregate data and added our own ideas to create a good research article. As a result of many studies carried out in a short time, detailed information was obtained about the entry of the virus into the cell and its cellular cycle. Although vaccination studies are about to come to an end, we do not yet have an agent that provides a definitive treatment that will facilitate millions of people's lives. At this point, humanity needs detailed genetic research, especially on cellular interactions. Because although we think that we are in control of the subject, science is a field that changes daily, and new data are added to it. It is evident that we need marginal ideas for a virus that has affected the whole world, can easily be transmitted by respiratory and droplets, and has destroyed everyday life. This research aims to examine the data we have so far in detail on all of the topics we have mentioned and try to make some suggestions within our knowledge.

Vetiver root is a commodity that is tolerant enough to be planted in salty soils with a certain level of salinity. One approach to increase oxidative stress tolerance that will increase the enzyme substrate at the cellular level is ascorbic acid. The purpose of this study was to increase students' knowledge about how the response of vetiver seed (Vetiveria zizanioides) growth in salt soils to ascorbic acid through practical field learning in plant physiology courses. This research was conducted using a factorial randomized block design (RBD), the first factor was the salinity (S), namely S0: 0 dsm-1 and S1: 4 dsm-1. The second factor was ascorbic acid (A), namely A0: without treatment, A1: 50 ppm, A2: 100 ppm and A3: 150 ppm. There were 8 treatment combinations that were repeated 3 times resulting in 24 experimental units. The parameters observed were plant height, number of leaves and number of stomata. In this study, ascorbic acid had a significant effect on plant height, number of leaves and number of stomata. Saline soil had a significant effect on the number of stomata parameters. No interactions for all parameters were observed.

Although aging is a strong and independent predictor of cardiovascular disease, an effective method to evaluate the degree of aging has yet to be reported. Here, we present a straightforward approach to determine aging in a rapid and quantitative manner, using a microfluidic-based electrochemical sensor equipped with a flexible membrane actuator. The flexible membrane in the sensor captures single endothelial cells of various ages (3, 4, and 18 month-old) under different cell capturing pressures (250, 300, and 350 kPa). The sensing electrodes in the sensor measure electrochemical impedance spectra of each cell group in terms of resistance and reactance. An optimal condition showing significant differences in resistance and reactance between different age groups was experimentally determined at a frequency of 1 MHz and a pressure of 350 kPa (p

HIGHLIGHTS A comprehensive review of increasing electromagnetic field radiation (EMFR) impacts on building occupant health Scientific evidence identifying adverse effects of EMFR Governments and public health agencies are creating regulations to reduce EMFR exposure National and international regulations with respect to the thresholds they set to protect human health Provides a robust foundation for researchers to use in additional studies of EMFR impacts in various built environment scenarios ABSTRACT A significant share of the technology that has emerged over the past several decades produces electromagnetic field (EMFR) radiation. Communications devices, household appliances, industrial equipment, and medical equipment and devices all produce EMFR with a variety of frequencies, strengths, and ranges. Some EMFR, such as Extremely Low Frequency (ELF), Radio Frequency (RF), and Ionizing Range (IR) radiation have been shown to have harmful effects on human health. Depending on the frequency and strength of the radiation, EMFR can have health effects at the cellular level as well as at brain, nervous, and cardiovascular levels. Health authorities have enacted regulations locally and globally to set critical values to limit the adverse effects of EMFR. By introducing a more comprehensive field of EMFR study and practice, architects and designers can design for a safer electromagnetic (EM) indoor environment, and, as building and construction specialists, will be able to monitor and reduce EM radiation. This paper identifies the nature of EMFR in the built environment, the various EMFR sources, and its human health effects. It addresses European and US regulations for EMFR in buildings and provides a preliminary action plan. The challenges of developing measurement protocols for the various EMFR frequency ranges and determining the effects of EMFR on building occupants are discussed. This paper argues that a mature method for measuring EMFR in building environments and linking these measurements to human health impacts will foster occupant health and lead to the adequate development of safeguards for occupants of buildings in future research.

Targeted Auger Therapy represents great potential for the therapy of diseases which require a high degree of selectivity on the cellular level (e.g. for therapy of metastatic cancers). Due to their high Linear Energy Transfer (LET), Auger emitters, combined with selective biological systems which enable delivery of radionuclides close to the DNA of the targeting cell, can be extremely selective and powerful treatment tools. There are two main aspects associated with the development of efficient radiopharmaceuticals based on Auger Emitters: a) the availability of suitable Auger-emitting radionuclides for therapy and b) the design of targeting vectors which can deliver Auger emitters into/close to the nucleus. In the present review, we address the first aspect by defining important parameters for the selection of radionuclides for application to Targeted Auger Therapy and form a categorized list of the most promising radionuclides, their possible production routes, and their use in the synthesis of radiopharmaceuticals.

Flow cytometry (FCM) is a sophisticated technique that works on the principle of light scattering and fluorescence emission by the specific fluorescent probe-labeled cells as they pass through a laser beam. It offers several unique advantages as it allows fast, relatively quantitative, multiparametric analysis of cell populations at the single cell level. In addition, it also enables physical sorting of the cells to separate the subpopulations based on different parameters. In this constantly evolving field, innovative technologies such as imaging FCM, mass cytometry and Raman FCM are being developed in order to address limitations of traditional FCM. This review explains the general principles, main applications and recent advances in the field of FCM.

Genetically identical cells in a tissue can respond differently to perturbations in their environment or “stress.” Such stresses can be physicochemical, mechanical, or infectious or may come from competition with other cells in the tissue. Here, I discuss how the varying responses to stress influence the decision of a cell to repair or die, and how one cell's response can have effects on surrounding cells. Such responses control the health and fitness of single cells and how they compete with other genetically identical cells. See related article on p. 129

The article presents the current state of knowledge on the differences between the sexes in the contraction features of motor units, motor and sensory innervation of skeletal muscles, features of motor neurons, proportions of contractile proteins and muscle fibers of various types, and morphometric features of muscles and muscle fibers. These studies, conducted at the cellular level, on animals, allow to explain the foundations of significant differences in the motor conditions of women and men.

Rozważania pierwszej części pracy dotyczą składu ciała człowieka, rozpatrywanego na pięciu różnych poziomach: A) poziom pojedynczych atomów, poprzez B) poziom związków chemicznych, C) skład ciała uwzględniający istnienie błony komórkowej (poziom komórkowy) i D) skład tkankowo-narządowy (poziom tkankowy), aż po E) ocenę ustroju jako całości. Szczególnie dużo uwagi w rozważaniach tych poświęcono mięśniom szkieletowym, w tym czynnikom warunkującym ich siłę i moc. W dalszych częściach pracy przedstawiono proces starzenia się organizmu człowieka, w tym starzenie się mięśni szkieletowych, z uwzględnieniem zjawiska otyłości sarkopenicznej oraz jej negatywnych skutków dla zdrowia człowieka. Omówiono również zjawisko starzenia się mięśni szkieletowych sportowców w przebiegu ich kariery sportowej jak i po jej zakończeniu. W końcowej części pracy przedstawiono dowody na korzystne oddziaływanie aktywności fizycznej / treningu na organizm osób w starszym wieku, nawet po przekroczeniu 100 roku życia.

Understanding the metabolic interactions between bacteria in natural habitat at the single-cell level and the contribution of individual cell to their functions is essential for exploring the dark matter of uncultured bacteria. The combination of Raman-activated cell sorting (RACS) and single-cell Raman spectra (SCRS) with unique fingerprint characteristics makes it possible for research in the field of microbiology to enter the single cell era. This review presents an overview of current knowledge about the research progress of recognition and assessment of single bacterium cell based on RACS and further research perspectives. We first systematically summarize the label-free and non-destructive RACS strategies based on microfluidics, microdroplets, optical tweezers, and specially made substrates. The importance of RACS platforms in linking target cell genotype and phenotype is highlighted and the approaches mentioned in this paper for distinguishing single-cell phenotype include surface-enhanced Raman scattering (SERS), biomarkers, stable isotope probing (SIP), and machine learning. Finally, the prospects and challenges of RACS in exploring the world of unknown microorganisms are discussed. KEY POINTS: • Analysis of single bacteria is essential for further understanding of the microbiological world. • Raman-activated cell sorting (RACS) systems are significant protocol for characterizing phenotypes and genotypes of individual bacteria.

Background: Allometric scaling is a well-known research tool used for the metabolic rates of organisms. It measures the living systems with fractal physiology. The metabolic rate versus the mass of the living species has a definite scaling and behaves like a four-dimensional phenomenon. The extended investigations focus on the mass-dependence of the various physiological parameters. Objective: Proving the length of vascularization is the scaling parameter instead of mass in allometric relation. Method: The description of the energy balance of the ontogenic growth of the tumor is an extended cell-death parameter for studying the mass balance at the cellular level. Results: It is shown that when a malignant cellular cluster tries to maximize its metabolic rate, it changes its allometric scaling exponent. A growth description could follow the heterogenic development of the tumor. The mass in the allometric scaling could be replaced by the average length of the circulatory system in each case. Conclusion: According to this concept, the dependence of the mass in allometric scaling is replaced with a more fundamental parameter, the length character of the circulatory system. The introduced scaling parameter has primary importance in cancer development, where the elongation of the circulatory length by angiogenesis is in significant demand.

Fluorescence-guided surgery provides surgeons with improved visualization of tumor tissue in the operating room to allow for maximal safe resection of brain tumors. Multiple fluorescent agents have been studied for fluorescence-guided surgery. Both nontargeted and targeted fluorescent agents are currently being used for glioblastoma multiforme visualization and resection. Fluorescence detection in the visible light or near infrared spectrum is possible. Visualization device advancements have permitted greater detection of fluorescence down to the cellular level, which may provide even greater ability for the neurosurgeon to resect tumors.

Molecular imaging is a non-invasive process that enables the visualization, characterization, and quantitation of biological processes at the molecular and cellular level. With the emergence of theragnostic agents to diagnose and treat disease for personalized medicine there is a growing need for matched pairs of isotopes. Matched pairs offer the unique opportunity to obtain patient specific information from SPECT or PET diagnostic studies to quantitate in vivo function or receptor density to inform and tailor therapeutic treatment. There are several isotopes of arsenic that have emissions suitable for either or both diagnostic imaging and radiotherapy. Their half-lives are long enough to pair them with peptides and antibodies which take longer to reach maximum uptake to facilitate improved patient pharmacokinetics and dosimetry then can be obtained with shorter lived radionuclides. Arsenic-72 even offers availability from a generator that can be shipped to remote sites and thus enhances availability. Arsenic has a long history as a diagnostic agent, but until recently has suffered from limited availability, lack of suitable chelators, and concerns about toxicity have inhibited its use in nuclear medicine. However, new production methods and novel chelators are coming online and the use of radioarsenic in the pico and nanomolar scale is well below the limits associated with toxicity. This manuscript will review the production routes, separation chemistry, radiolabeling techniques and in vitro/in vivo studies of three medically relevant isotopes of arsenic (arsenic-74, arsenic-72, and arsenic-77).

Interdisciplinary researches have been well appraised in recent times and has to a greater extent helped the CAM community to develop scientific validation for the safety and efficacy of traditional and indigenous formulations. Bhasmas or incinerated organometallic preparations are highly acclaimed with regard to therapeutic efficacy among the Ayurvedic practitioners and has thus, created an arena for various unvalidated and unscientific claims. Bringing order in chaos is a basic purpose that interdisciplinary researches serves to. Addressing the various concerns on the lack of scientific validation on chemical characterization and preclinical studies at cellular level, an attempt is being made to develop a biological safety profile protocol for bhasmas using advanced instrumentation techniques and in vitro methods.